Document ev0oj6Gy2bgdw1Xvz7xrq80J9

IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION 1 CECIL SCOTT, ET AL v. MONSANTO COMPANY ] ] No. B-84-1103-CA ] EXHIBITS TO VIDEOTAPE DEPOSITION OF DR. GEORGE LEVINSKAS May 28, 1987 1300 Post Oak Boulevard Houston, Texas Wanda G. Kuhn, Court Reporter Nell McCallum a Associates Inc. 2900 Smith, Suite 104 Houston, Texas 77006 (713) 523-3767 NELL MC CALLUM & ASSOCIATES, INC. HARTOLDMON0023204 OMtrt ii s DMEH C'*: Toxicology Section/St. Louis tcoJOtvjoerr^LOCATioN) SPECIAL (TYPE OF REPORT) REPORT in N i-i CO -g (iVrI pi (VJ O i-t 3-5t a-isi aa xCu* *-* caQj <ri U. in- oa. o Xca a. tXu <n o > -Ci _O ix-i > CJ JDMOMt <t- "} u ott. Ot 0V0. >HvoO ONN UO UII H xa Sj3 O .. <P o o CM x co z d z O z bfc > Ul cu -1 <(* REPORT NO.: MSL-2 002 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: TOXICITY OF AROCLOR PRODUCTS 1242,; 1254 AND 1260 TO THE LIVER OF ALBINO-RATS AUTHORS: George J. Levinskas, Ph.D. asstract: This report is a tabulation of the results of microscopic evaluation of liver sections from rats fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 1260 for two years. SCM 058930 000007 HARTOLDMON0023205 DISTRIBUTION COPY NUMBER 1 - Reports.Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEHLibrary, G2WA 5 - T3MEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H. Craddock, A2SA 8 - G.J. Leviriskas, G2WF 9 - J.G. Nassi'f, E2ND '' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: ______________REPORTS LIBRARY, R2C ____________ If you transfer it to anyone else, please let your librarian know, so the records can be changed. R.IOII(C) (RCV. l/M) OOOCG'^cm 05893 L HARTOLDMON0023206 INTRODUCTION The polychlorinated biphenyls (PCBs) comprise a family of compounds of variable chlorine content. PCBs manufactured by Monsanto (tradenamed "Aroclor") bear a four digit number, the '12' indicating biphenyl and the last two digits indicating the chlorine content by weight percent. Since the chlorine atoms are randomly distributed among the 10 ring positions available for substitution, each material is a mixture of isomers. In 1969, Monsanto sponsored a series of animal studies at Industrial BIO-TEST Laboratories in Northbrook, Illinois, on Aroclor 1242, 1254, and 1260 for the assessment of the health and environmental hazards of these materials.. This series consisted of 2-year chronic feeding studies to rats and dogs, a 3-generation rat reproduction study, a rat teratology study, a dominant lethal mutagenic study in mice and a toxicity/reproduction study in chickens on each of the 3 Aroclor products. Even though no such action was contemplated, such a broad battery of tests would have been adequate to support Food Additive Petitions for each of these materials. These studies were initiated by reports that PCBs had been detected in the environment. A report (Nelson, 1972b) by a Panel on Hazardous Trace Substances of an Ad Hoc Committee on Environmental Health Research covers the development of information on the environmental and biological effects of PCBs. There have been subsequent literature reviews which need not be recapitulated here [DHEW, 1978; EPA, 1976; IARC, 1978; NAS, 1979; Roberts, et al. (1978)]. Starting in 1969, while these studies still were in progress, information regarding them was made available to various government groups.1 Subsequently, data from these studies were presented at a conference '. - i- 000CG3 SCM 058932 HARTOLDMON0023207 sponsored by the National Institute of Environmental Health Sciences at Rougemont, N.C. on December 20-21, 1971, but the account of these studies was omitted from the published proceedings (Keplinger, et al., 1972; Nelson, 1972a). Subsequently, it was reported that female Sherman strain rats developed hepatocellular carcinomas when fed Aroclor 12602 from Lot No. AK-3; the same lot used for the earlier Monsanto study with this material. As a result, Monsanto requested the contract laboratory's pathologists to review livers from all available rats from the 3 Aroclor studies. That review (which could have included new sections of liver from animals examined previously in addition to sections from animals not examined previously) was presented in the reports of Gordon and Richter (1975a,b,c). In November 1975, reports containing evaluations of liver sections from the 2-year rat feeding studies (Gordon and Richter, 1975a,b,c) and the results of an independent evaluation of the same liver sections (Pour, 1975) were presented to and discussed with several federal regulatory agencies3. Later that month, a summary of data from all of the studies was presented at the National Conference on Polychlorinated Biphenyls sponsored by the Environmental Protection Agency and held in Chicago on November 19-21, 1975 (Calandra, 1976). ' Only summaries of data from these and other toxicity studies conducted over the years have been published (Jenkins, et al.. 1972; Keplinger, et al., 1971; Monsanto, 1980). Knowledge of these efforts may have prompted the statement in a recent article that "...Monsanto, whose reaction to the possibility that polychlorinated biphenyls (PCBs) might be an ecological disaster was a classic of how such issues should be handled, says Ford4. Monsanto began to investigate the dangers as soon as they were 2 - OOOCGO SCH 059,33 HARTOLDMON0023208 seriously voiced. It insisted on keeping an open mind about them. As soon as evidence appeared that there was a strong chance PCBs were a hazard, it published the results of its investigations, admitting the danger. It thus established a reputation of honesty even among the environmentalists.'1 (Clutterbuck 1981). At a later meeting in Bethesda, MD.S, pathologists selected and reviewed some liver slides from the Monsanto-sponsored and Kimbrough studies. The pathologists differed in the terminology used to classify the lesions. They did conclude that the general incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) were greater in the rats from the study subsequently published by Kimbrough, et al. (1975). No carcinomas were seen in the Monsanto-sponsored studies. It was noted that either the strain of rat or sex could have accounted for the differences. The spontaneous incidence of hyperplastic or neoplastic liver lesions in the Sherman strain rat was unknown, and the occurrence of such lesions appears to be higher in female rats and mice. The different diagnoses were discussed with Dr. Philippe Shubik of the Eppley Institute for Research in Cancer at the University of Nebraska. Dr. Shubik suggested that the liver sections from these studies could be reviewed by Dr. Parvis Pour. This was done. This publication is an effort to make readily available information in the Gordon and Richter reports (1975a,b,c) which are only summarized in the literature (Calandra, 1976). 000-01 - 3 SCH 058934 HARTOLDMON0023209 METHODS It appears that the Threshold Limit Value of 0.5 mg/m3 for chlorobiphenyl with 54% chlorine (ACGIH, 1969) was used to estimate suitable dose levels for the rat feeding studies. For that purpose the following assumptions were made: if the ambient air concentration of PCBs is 0.5 mg/m3, and if all of the inhaled PCBs are absorbed, and if 15 m3 of air are inhaled during the working day, a person would receive a PCB dose of 7.5 mg/day. The corresponding dosage would be approximately 0.1 mg/kg/day for a 70-kilogram person. Consequently, dosages of 0.1, 1, and 10 mg/kg/day were decided upon, and the dietary concentrations were set at 1, 10, and 100 ppm. As it turned out, the assumptions used to set dosages for the feeding study resulted in the low dose level rats receiving a dosage that was 100 times higher than the 0.001 mg/kg/day which FDA later calculated as allowable for protracted ingestion based on human data (FDA, 1973). For the chronic toxicity study in rats6, weanling animals of the Charles River CD strain7 were divided into a control group and 9 treatment groups, each consisting of 50 males and 50 females, with 3 treatment groups assigned to each of the 3 Aroclor products studied (1242, 1254, and 1260). Not all of these animals started at the same time. After about 2 months on test, additional groups of males and females were assigned to each test diet and the controls. These animals were added to allow a sufficient number of animals for sacrifices at 3, 6 and 12 months to provide some information prior to the completion of the 2-year study period. The numbering sequence 4- 0 j'-j' SCM 058935 HARTOLDMON0023210 and the designation of some animals as "Extra" suggests that additional animals were placed on test. Groups of 5 males and 5 females were scheduled to be sacrificed after 3, 6, and 12 months of feeding, and survivors were to be sacrificed at the end of the test period.* Animals were to be given a gross autopsy and tissues were to be preserved for possible histologic examination. Tumors or lesions suggestive of tumors were to be examined. RESULTS Data from the Gordon and Richter reports (1975a,b,c) on liver slides are shown in Tables 1, 2, and 3 for Aroclor 1242, 1254, and 1260, respectively. While the text of the reports contained comments specific to the particular Aroclor, they also contained similar comments such as "There is evidence of a chemical effect on the liver..........This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoma" and "In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study". The reports also stated that "The other treatment-related lesions reported are regarded as degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotrans formation of the test material" and "In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report..........No hepatocellular carcinomas were observed". 5- OSCHV,05:S936 HARTOLDMONOQ23211 With respect to Aroclor 1254, it was noted that "One liver tumor (a hepatoma) previously reported in a T-II animal (No. 445) was re-classified as nodular hyperplasia" (Gordon and Richter, 1975a)1. DISCUSSION The initial reports of these studies concluded that the target organ was the liver for each Aroclor product. This was confirmed by the second evaluation of liver slides (Gordon and Richter, 1975a,b,c). These alterations were slow to develop, their incidence being related to both dose level and duration of treatment. Since there were increased incidences of vacuolar changes in the cytoplasm of the hepatocytes and focal hypertrophy at all dose levels for all 3 Aroclor products at the 24-month sacrifice, a "no-effect" level was not established for liver effects. With respect to the slides from Industrial BIO-TEST, Pour (1975) concluded that Aroclor 1242, 1254, and 1260 "showed a dose-dependent hepatotoxic effect, characterized by degenerative and regenerative processes. With one exception, all lesions were considered non-neoplastic. Structures similar to cholangiocarcinomas and hepatomas were found in one rat. However, the possibility of metastases of a carcinoma into the liver has been also considered." In the report, he noted one lesion which seemed to "represent a metastatic tumor (probably a mammary gland carcinoma of the same animal from which ...(the particular liver section]... was submitted), rather than a cholangiocarcinoma". This occurred in an animal fed 100 ppm of Aroclor 1254. The contract laboratory pathologists diagnosed the presence of mammary tumor metastases in the liver of this rat (Gordon and Richter, 1975b).11 . -6- . 000-01 so, 05893-1 HARTOLDMONOQ23212 SUMMARY and CONCLUSIONS Groups of male and female rats were fed diets containing either 1, 10, or 100 ppm of one of 3 Aroclor products, 1242, 1254, or 1260, for 2 years. Focal hypertrophy of the liver occurred at 3, 3, and 12 months for rats fed Aroclor 1260, 1254, and 1242, respectively. Focal hypertrophy was not seen in livers of control animals. At the 24-month sacrifice, livers of animals from all dose levels of the 3 Aroclor products had an increased incidence of vacuolar changes and focal hypertrophy. Livers of some animals fed 100 ppm of each Aroclor also had benign liver tumors (hepatoma or cholangiohepatoma). No hepatocellular carcinomas were observed. GOQC.O.j 7 - SCM 058938 HARTOLDMONOQ23213 Footnotes Dates of initial correspondence and contacts. October 3, 1969. E.P. Wheeler to A.R. Glasgow, Division of Pesticides. Food and Drug Administration. April 8, 1970. R.E. Kelly to H. Blumenthal, Petitions Review Branch, Food and Drug Administration. ' April 22, 1970. E.P. Wheeler to E.J. Burger, Jr., Office of Science and Technology. June 1, 1970. E.P. Wheeler to H.E. Stokinger, Bureau of Occupational Safety and Health. R.D. Kimbrough, personal communication. November 13-14, 1975. Council on Environmental Quality. Environmental Protection Agency. Food and Drug Administration. House Subcommittee Manpower, Compensation, Health and Safety. National Cancer Institute. National Institute for Environmental Health Sciences. National Institute for Occupational Safety and Health. Dr. David Ford of Bath University. At National Cancer Institute on January 21, 1975. Present were D.E. Gordon, R.D. Kimbrough, G.J. Levinskas, R.A. Squire, W.R. Richter. _ . Since the events described earlier, the validity of many toxicity studies conducted by Industrial BIO-TEST Laboratories has been challenged. Therefore, the available raw data supplied by Industrial BIO-TEST was reviewed to determine whether this study could be validated. The review showed that the data bases (including the lack of a protocol) were insufficient for a complete validation of the study. It was decided to focus on presenting the primary liver effects reported by Gordon and Richter (1975a,b,c). Therefore, a complete audit was not undertaken. Available records were examined for a determination that the animals were placed on test and of their ultimate fate. Necropsy and microscopic reports were also examined for findings pertaining to livers of those animals. Significant discrepancies which were found between data in Tables 1, 2, and 3 and the data base for the Gordon and Richter reports are noted in this report. On some records, the labels Aroclor 1242 and Aroclor 1260 are interchanged as determined by a check of the animal numbers., Charles River Breeding Laboratories, North Wilmington, Massachusetts. Animals sacrificed at 6 and 12 months were placed on test about 2 months after the first group. Those sacrifice periods are sometimes labeled 8 and 14 months, respectively, which corresponds to the calendar time from the start of the test but overstates the time on test for those groups. OnQ-36 - 8 058^ sort HARTOLDMONOQ23214 9. As a result of the examination described in footnote 6, a few animals were reassigned to other dose levels on basis of assigned numbers. Three with no recorded liver lesions were deleted from the 24-month listing because of short duration on test: 14 months at 100 ppm Aroclor 1242, 15 months at 10 ppm Aroclor 1254 and 13 months at 100 ppm Aroclor 1254. Therefore, numbers in Tables vary slightly from those in reports of Gordon and Richter (1975a,b,c). 10. That change and comments in the footnotes to the Tables were noted ' during the examination described in footnote 6. In addition, the following also were noted during the examination. Aroclor 1254 - 100 ppm animal marked "Extra3" with diagnosis of hepatoma in record of examination for original Industrial BIO-TEST report. Animal not listed in Gordon and Richter report. . - 100 ppm animal no. 542 with gross autopsy notation that liver appears tumorous and white foci has no record of examination. Aroclor 1260 - 100 ppm animal no. 293 with gross autopsy notation of . tumor on liver has no record of examination. In some instances, diagnoses were crossed out on the available records. These were included in the Tables. Crossed out diagnoses for hepatoma or cholangiohepatoma are noted in the footnotes to the Tables. - 11. Dr. Pour's report does not include the following liver sections noted by discrepancies between his tabulation and the Gordon and Richter reports. 1 from control at 12 month sacrifice, 5 from 100 ppm Aroclor 1242 at 24 months, 1 from 100 ppm Aroclor 1260 at 3 months. None of these animals were reported as having hepatomas or cholangiohepatomas in the Gordon and Richter reports. 000C07 SCH 058940 HARTOLDMON0023215 References 1. ACGIH (1969). Threshold Limit Values of Airborne Contaminants. American Conference of Governmental Industrial Hygienists. Cincinnati. 2. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. In: National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. EPA Report No. 560/6-75-004. March. - NTIS PB-253 248. pp.35-42. 3. Clutterback, D. (1981). What causes environmental conflict? New Scientist 90, 176-177. 4. DHEW (1978). Final Report of the Subcommittee on Health Effects of PCBs and PBBs. Environ. Health Perspect., 24, 129-198. 5. EPA (1976). National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. U.S. Environmental Protection Agency. Washington, D.C. EPA-560/6-75-004, March. NTIS PB-253 248. 6. FDA (1973). Polychlorinated biphenyls (PCB's). Contamination of animal feeds, foods and food-packaging materials. Fed. Regis., July 6, 38, 18096-18103. 7. Gordon, D.E. and Richter, W.R. (1975a). Two-year chronic and toxicity study with Aroclor 1242 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 8. Gordon, D.E. and Richter, W.R. (1975b). Two-year chronic oral toxicity study with Aroclor 1254 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 9. Gordon, D.E. and Richter, W.R. (1975c). Two-year chronic oral toxicity study with Aroclor 1260 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 10. IARC (1978). Polychlorinated biphenyls. IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. 18, 43-103. International Agency for Research on Cancer. Lyon, France. October. 11. Jenkins, D.H., Keplinger, M.L., Fancher, O.E., Wheeler, E.P. and Calandra, J.C. (1972). Reproductive effects of polychlorinated biphenyls in white leghorn chickens. Toxicol. Appl. Pharmacol. 22, 316. 12. Keplinger, M.L., Fancher, O.E., Calandra, J.C. (1971). Toxicologic studies with polychlorinated biphenyls. Toxicol. Appl. Pharmacol. 19, 402-403. 00QC33 SCM 0589*1 HARTOLDMONOQ23216 13. Keplinger, M.L., Fancher, O.E., Calandra, J.C., et al. (1972). Toxicological studies with polychlorinated biphenyls. Read at PCB Conference, Quail Roost Conference Center, Rougemont, North Carolina, 20-21 December 1971. Cited in Polychlorinated Biphenyls Environmental Impact. A Review by the Panel on Hazardous Trace Substances. March 1972. Environ. Res. 5 , 249-362. 14. Kimbrough, R.D., Squire, R.A., Linder, R.E., Strandberg, J.D., ' Montali, R.J., and Burse, V.W. (1975). Induction of liver tumors in Sherman Strain female rats by polychlorinated biphenyl Aroclor 1260. J. Natl. Cancer Inst. 55, 1453-1459. 15. Monsanto Company (1980). Polychlorinated biphenyls. PCB's. A report on Uses, Environmental and Health Effects and Disposal. 16. NAS (1979). Polychlorinated biphenyls. National Academy of Sciences. Washington, D.C. 17. Nelson, N. (1972a). Comments on research needs. Environ. Health Perspect., 1, 181-185. 18. Nelson, N. (1972b). Chairman, Panel on Hazardous Trace Substances. Polychlorinated biphenyls - environmental impact. Environ. Res. 5, 249-362. 19. Pour, P. (1975). Histopathological reevaluation of livers for rats treated with Aroclor. August 1 (unpublished observations). 20. Roberts, J.R., Rodgers, D.W., Bailey, J.A., Rorke, M.A. (1978). Polychlorinated Biphenyls: Biological criteria for an assessment of their effects on environmental quality. National Research Council of Canada. Ottawa. Publication No. NRCC 16077. SCH 058942 HARTOLDMON0023217 Sacrifice Interval Diet Level (ppm) Table 1 Aroclor 1242: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cho1angiohepa toma 10 8 10 9 202 010 000 000 000 000 000 000 1 0 0 0 0 1 0 0 10 10 8 9 1 00 1 10 030 000 000 000 000 000 0 0 0 0 0 0 0 0 10 10 10 9 1 24 000 00 1 000 000 101 000 000 0 0 0 1 0 0 0 0 23 32 29 19 178 111 100 023 111 533 000 000 9 0 0 8 8 3 3b 1C * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group haa animals dead at 19, 22, 22, 23, 23 months. 10 ppm group has animals dead at 23, 23 months. ,t100 ppm group has animals dead at 22, 22, 22 months and 2 marked "Extra". ^ Includes 1 animal without record of examination in original IBT report. Not listed as hepatoma in worksheets for ^ Gordon and Richter report but appears and was crossed out on typed tabulation for anioial marked "Extra". O for other 2 animals do not appear in records of examinations for original IBT report. O'- . t-)C Diagnosis does not appear in records of examinations for original IBT report. Diagnoses SCM 0 5 8 9 4 3 HARTOLDMON0023218 Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cholangiohepatoma 10 10 10 10 233 01 1 000 000 000 000 000 000 1 0 1 1 0 0 0 0 10 10 10 10 1 00 1 20 000 000 000 000 000 000 0 2 2 4 0 0 0 0 10 10 10 10 1 14 000 002 004 000 111 000 000 1 1 1 2 0 0 0 0 23 30 26 26 1 7 9 13 131 1 120 1 0 3 4 14 1 0 3 14 563 000 4 4b 000 2C * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 22, 22 months, 1 marked "Extra" and 1 other for which date of death ia not recorded. 10 ppm group has animals dead at 22, 22, 22 months. ,,100 ppm group has animals dead at 23, 23 months and 9 marked "Extra". b Includes 2 animals marked "Extra". Diagnoses do not appear in records of examination for original IBT reports. o C3c Both animals marked "Extra" with same designation. 1 without apparent record of examination for original IBT report. Diagnosis for other animal does not appear in records of examinations for original IBT report. b ' '1 H- HARTOLDMON0023219 iJ 'JD O ijV J *J1> *4 00 Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats Sacrifice Interval Diet Level (ppm) 3 Honths 0 1 10 100 6 1Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined 10 10 iob 10 10 6 Findings Vacuolar change Focal necrosis 202 3 0 0 4C 0 11 10 .Focal lymphoid infiltration Focal hypertrophy hepatocytes 000 000 0 2 02 00 Nodular hyperplasia 000 0 00 Ductular hyperplasia 00 1 0 00 Hepatoma Cholangiohepatoma 000 000 0 0 00 00 *. Control group has animal dead at 19 months and 2 marked "Extra". 5 2 ld 2 0 0 0 0 0 9 6 0 0 3 0 0 0 0 10 9 10 9 1 25 000 0 00 000 000 1 10 000 000 3 0 0 4 1 2 0 0 23 26 25 25 157 9 143 6 101 0 0 3 13 9 107 6 5 6 7 12 0 0 le 000 48. * 1 ppm group has animals dead at 20, 22, 22 months. 10 ppm group has animals dead at 19, 22, 22 months and 1 marked "Extra". 100 ppm group has animals dead at 22, 22 months. ** Includes 1 marked "Extra". c Worksheets show listings under this diagnosis with arrow pointing to Vacuolar change colusw. d,Date of death of this animal not recorded. e No record of examination for original IBT report. Listed on worksheets for Gordon and Richter report with diagnosis crossed out. * Includes 2animals without record of examination for original IBT report. Diagnoses for other 5 animals do not appear in q records ofexaminations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter j reports. O Includes one animal with both diagnoses. Hepatoma is 1 of 2 crossed out (superscript f). O Includes 3animals without record of examinationfor original IBT report. Diagnosis for other animal does not appear in tO records ofexaminations for original IBT report. 2 of these diagnoses crossed out on worksheets for Cordon and Richter reports. HARTOLDMON0023220 V E X H I B I T ft HARTOLDMON0023221 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT. ET AL VS. MONSANTO COMPANY CIVIL ACTION NO B-84-1103-CA PLAINTIFFS' NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its attorneys of record Mr. Robert A. Hall, Mr. Jonathan B. Shoebotham, Woodard, Hall & Primm, 4700 Texas Commerce Tower, Houston, Texas 77002; Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3709, Beaumont, Texas 77704; Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, . Birmingham, Alabama 35203; and Mr. Michael R. Fruehwald, Mr. Michael Rosiello, and Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Building, 11 South Meridian Street, Indianapolis, Indiana 46204. Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give NOTICE to the Defendant that the oral deposition of MONSANTO COMPANY, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the representative(s) designated by the 2 DEPOSITION f EXHIBIT HARTOLDMON0023222 Defendant pursuant to Rule 30(b) (6) of the Federal Rules of Civil Procedure. Plaintiffs request that Defendant designate that current employee of Defendant having held the highest position of any held by a current employee in Monsanto Company's toxicology department during the period 1965 through 1975. Plaintiffs also request that Defendant designate that representative knowledgeable of the documents and subject matter herein below enumerated. 1) Any document (s) which would state or disclose the following: a) when Monsanto first began using the services of IBT; b) when Monsanto first began using the services of IBT in connection with Monsanto's PCB products; c) the dollars spent by Monsanto with IBT (i) each year that Monsanto used IBT's services, (ii) on all PCB related studies, and (iii) totally, from the date Monsanto first began using the services of IBT. 2) All correspondence from IBT to Dr. Paul L. Wright while Dr. Wright was an employee of Monsanto; 3) All correspondence by Monsanto to Paul L. Wright or from Paul Wright to Monsanto; 4) Any document stating, reflecting or referring to the policies and procedures of Monsanto's Toxicology Department for the years 1965 through 19 80; 5) Any document stating, reflecting or referring to the current standards, procedures and policies of Monsanto's Toxicology Department; 00QCQ1I -2- HARTOLDMON0023223 6) The results of any IBT PCB test redone by or for Monsanto Company; 7) Any investigation or background file on Dr. Renata Kimbrough; 8) Any correspondence to or from Dr. William Ribelin; 9) Any report, analysis or investigation of IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products (this request includes copies of government reports; independent investigations; internal IBT investigations; Monsanto authored or sponsored investigations; and, investigations by industry groups, bodies or authorities or other private or public body) (IBT includes all of its employees and consultants). 10) Any document which would in any way reflect, relate or pertain to a change by or on behalf of Monsanto to the results (interim or final) of any test of a Monsanto product by IBT; 11) Any document which would in any way reflect, relate or pertain to a change by or on behalf of Monsanto to the results (interim or final) of any test of a Monsanto PCB product by IBT; 12) Any document which would in any way reflect, relate or pertain to a suggestion by Monsanto regarding the outcome of a test to be conducted by IBT for Monsanto on any Monsanto product; 13) Any document which would in any way reflect, relate or pertain to a suggestion by Monsanto regarding the outcome of a test to be conducted by IBT for Monsanto on any Monsanto PCB product. This deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, G00C03 -3- HARTOLDMON0023224 23rd Floor, Houston, Texas 77056 beginning at 9:00 a.m. on Saturday, May 23, 1987 and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together with the documents produced at this deposition. You are invited to attend and dross-examine. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers, financial statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. "PCB" means polychlorinated biphenyl and its contaminants and derivatives. ooooo** -4HARTOLDMON0023225 Respectfully submitted, By:______________________ Michael A. Pohl OF COUNSEL: David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Boulevard Allied Bank Tower, 23rd Floor Houston, Texas 77056 (713) 623-8800 REAUD, MORGAN & QUINN 909 Laurel Avenue Beaumont, Texas 77701 (409) 838-9941 Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania (412) 471-3980 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 . 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. CERTIFICATE OF SERVICE QQQCOn -5HARTOLDMONOQ23226 I hereby certify that on the ______ day of , 1987 , a true and correct copy of the above Notice of Intent to Take Oral Deposition and Subpoena Duces Tecum was served upon counsel of record either by messenger or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3708 Beaumont, Texas 77704 Mr. John M. Johnson Mr. Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruehwald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 1313 Merchants Bank Building 11 South Meridian Street Indianapolis, Indiana 46204 depntc02/txt8 65 51 Michael A. Pohl -6- OOQCCC HARTOLDMON0023227 u ixx V \ I B I T Ii ! HARTOLDMON0023228 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT, et al Plaintiffs V. MONSANTO COMPANY C. A. NO. B-84-1103-CA Defendant PLAINTIFFS * NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its counsel of record, Mr. Jonathan B. Shoebotham and Mr. Robert A. Hall, Woodard, Hall <5 Priirnn, 4700 Texas Commerce Tower, Houston, Texas 77002 Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt <5 Crawford, 624 Petroleum Bldg., P. 0. Box 3708, Beaumont, Texas 77704 Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, Birmingham, Alabama 35203 Mr. Michael R. Fruewald, Mr. Michael Rosiello, Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Bldg. 11 South Meridian Street, Indianapolis, Indiana 46204 Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give notice to the Defendant that the oral deposition of Monsanto Company, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the representative( s) designated by the 000C07 HARTOLDMON0023229 Defendant pursuant to Rule 30(b)(6) of the Federal Rules of Civil Procedure and charged with the responsibility for maintaining custody of the documents listed below and such designated representative(s) shall testify as to matters known or reasonably available to the Defendant as well as the custody of the herein designated records and their maintenance by Defendant: 1. The complete personnel file of Paul L. Wright; 2. Any bonuses, payment dividends or other consideration other than normal salary paid by Monsanto or anyone on its behalf to Paul L. Wright or his designee/assignee; 3. Paul L. Wright's job evaluation and/or reviews; 4. Any payment to or for the account of attorneys hired by or for Paul L. Wright in connection with matters pertaining to his employment at Industrial Bio-Test Laboratories, Inc. ("IBT"). See United States v. Keplinger. 776 F.2d 678 (7th Cir. 1985); 5. Any correspondence to or from Paul L. Wright; 6. Paul L. Wright's job duties and responsibilities as an employee of Monsanto both before and after he was employed by IBT; and, 7. The matters identified in the below listed subpoena duces tecum. The deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, 23rd Floor, Houston, Texas 77056 beginning at 1:30 p.m. on Friday, May 8, 1987, and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together G0QC03 HARTOLDMON0023230 with the documents produced at this deposition. You are invited to attend and cross-examine. ' Pursuant to Rules 30(b)(5) and 34 of the Federal Rules of Civil Procedure, certain documents shall be produced by the designated representative(s) or custodian(s) of Monsanto Company (referred to herein as "Monsanto") at the commencement of the oral deposition or prior thereto by agreement of counsel. The originals and all drafts of the following requested documents shall be produced: 1. The complete personnel file of Paul L. Wright; 2. Any payment by Monsanto or anyone on its behalf to or for the benefit of Wright Paul L. Wright; 3. Any payment by or for Monsanto to attorneys for Paul L. Wright in connection with any criminal prosecution arising out of or in any way related to his employment at IBT; 4. All correspondence to or from Paul L. Wright; 5. All evaluations of Paul L. Wright's performance as an employee of Monsanto; 6. All documents pertaining to the termination of Paul L. Wright's Monsanto employment (both before being first, employed by IBT and as a result of or in connection with the criminal proceedings relating to IBT). See United States v. Keplinqer. 776 F.2d 678 (7th Cir. 1985); 7. All documents pertaining to Monsanto's efforts to monitor, supervise, keep abreast of or in any way observe the IBT criminal trial. See United States v. Keplinqer. 776 F.2d 678 (7th Cir. 1985); 8. Any report, analysis or investigation of IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products (this request includes copies of government reports; independent investigations; internal IBT investigations; Monsanto authored or sponsored investigations; -3- 0000013 HARTOLDMONOQ23231 and, investigations) by industry groups, bodies or authorities or other private or public body) (IBT includes all of its employees and consultants). Unless otherwise specified herein, the documents to be produced include all documents in the possession of Monsanto from the date of the first production or manufacture of PCBs in any form by Monsanto to the present time. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers, financial, statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. By:--------------------------------------------------------------JOSEPH C. BLANKS -4- ooo(;:o HARTOLDMON0023232 OF COUNSEL: REAUD, MORGAN & QUINN 909 Laurel Avenue Beaumont, Texas 77701 (409) 838-9941 Michael A. Pohl David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Blvd. Allied Bank Tower, 23rd Houston, Texas 77056 (713) 623-8800 Floor Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. -5- oooo;i HARTOLDMON0023233 CERTIFICATE OF SERVICE I hereby certify that on the ______ day of , 1987, a duplicate original of the foregoing Plaintiffs' Notice of Intention to Take Oral Deposition was filed with the Clerk of the Court for the Eastern District of Texas, Beaumont Division. I also certify that a true and correct copy of this Notice was served upon counsel of record on the ______ day of ___________ 1987, by pessenger delivery and/or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford P. 0. Box 3708 Beaumont, Texas 77704 Mr. John M. Johnson Mr. Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruewald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 1313 Merchants Bank Bldg. 11 South Meridian Street Indianapolis, Indiana 46204 MAP:j ar/scottmon/054 JOSEPH C. BLANKS -6ococ:^ HARTOLDMON0023234 UJXX fI V___ _ A I B I T ( I HARTOLDMON0023235 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT, ET AL. VS. MONSANTO COMPANY CIVIL ACTION NO. B-84-1103-CA PLAINTIFFS' NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its attorneys of record Mr. Robert A. Hall, Mr. Jonathan B. Shoebotham, Woodard, Hall & Primm, 4700 Texas Commerce Tower, Houston, Texas 77002; .' Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3709, Beaumont, Texas 77704; Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, Birmingham, Alabama 35203; and Mr. Michael R. Fruehwald, Mr. Michael Rosiello, and Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Building, 11 South Meridian Street, Indianapolis, Indiana 46204. Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give NOTICE to the Defendant that the oral deposition of the Manager of Toxicology of MONSANTO COMPANY, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the ' 2 DEPOSITION \ EXHIBIT 2l HARTOLDMON0023236 representative (s) designated by the Defendant pursuant to Rule 30(b) (6) of the Federal Rules of Civil Procedure and charged with the responsibility for maintaining custody of the documents listed below. The designated representative(s) shall testify as to: a) Matters known or reasonably available to the Defendants concerning the policies and procedures of Monsanto's Toxicology Department for the years 1965 through 1980; b) The current standards, procedures and policies of Monsanto's Toxicology Department; c) The duties and responsibilities of the Manager of Monsanto's Toxicology Department (i) 1965 through 1980 and (ii) currently; d) The organization of Monsanto's Toxicology Department (i) 1965 through 1980 and (ii) currently; e) The interrelationship between Monsanto's Toxicology Department and Monsanto's other departments and/or sections; f) The relationship between Monsanto's Toxicology Department and any outside testing facilities (i) 1965 through 1980 and (ii) currently; g) Any industry or government guidelines or regulations relating or pertaining to tests concerning the toxicological effects of Monsanto products on animals; h) Any participation by Monsanto's Toxicology Department in any effort by Monsanto to avoid or defer the ban of its PCB products; i) The day-to-day activities of the current Manager of Monsanto's Toxicology Department; and -2- 000CM HARTOLDMON0023237 j) The documents designated in the accompanying Subpoena Duces Tecum. This deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, 23rd Floor, Houston, Texas 77056 beginning at 9:00 a.m. on Saturday, May 23, 1987 and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together with the documents produced at this deposition. You are invited to attend and cross-examine. Pursuant to Rules 30 (b) (5) and 34 of the Federal Rules of Civil Procedure, certain documents shall be produced by the designated representative(s) MONSANTO COMPANY (referred to herein as "Monsanto") at the commencement of the oral deposition. The originals and all drafts of the following requested .documents shall be produced: 1) Any manual, compilation or other writing setting forth or reciting the policy or procedures for the Monsanto Toxicology Department from 1965 to date; 2) Any document pertaining to Monsanto's standards for the feeding or care of animals used in connection with the testing of Monsanto products; 3) Any document pertaining to the care, skill or precision to be used (a) by Monsanto and (b) by any outside/independent testing laboratory in gathering, recording, maintaining and/or -3- 0000:5 HARTOLDMON0023238 reporting of data generated by or from animal studies; 4) Any document comparing or contrasting the results of IBT Aroclor tests with tests conducted by (a) Monsanto, (b) others on Monsanto's behalf and/or (c) independent or government researchers; 5) Any document regarding any investigation or analysis of those Aroclor tests conducted by IBT for Monsanto; 6) Any document regarding or otherwise pertaining to trips (a) by Monsanto employees or representatives (i) to the offices of IBT (ii) to other locations where IBT representatives were met or (b) by IBT employees or representatives (i) to the offices of Monsanto or (ii) to locations where IBT and Monsanto employees or representatives met; 7) All literature regarding PCBs which was supplied or made available to Monsanto's experts, or consultants; 8) All Electrical Power Research Institute reports, articles, papers, etc., pertaining or relating to PCBs; 9) Any list or summary identifying (a) which IBT studies were redone by or for Monsanto and (b) the variances or similarities between the Monsanto tests conducted by IBT and those redone by or for Monsanto; 10) Any industry (and/or government) guidelines, standards or suggested guidelines/standards applicable to toxicological studies from 1960 to date (e.g. Aroclor studies on rats, beagle dogs, leghorn hens and/or fish) including any variations thereof from their inception to date; 11) Any document setting forth or otherwise stating Monsanto's standard of care regarding the following data in connection with toxicological tests (gathering, recording, maintaining); a) blood data; Q00C1G -4- HARTOLDMON0023239 b) urine analysis; c) body weight; d) feeding data; e) autopsy (procedures and results) ; f) viscera; g) microscopic analysis; h) organ analysis; i) death of test animals; j) substitution of animals; k) loss of animals due to escape or confusion of identity. 12) All correspondence between IBT and Dr. Paul Wright while Dr. Wright was employed by Monsanto; 13) Any document evidencing or suggesting that Monsanto corrected, changed, altered or otherwise amended (a) any report of a test performed by IBT for Monsanto and (b) any paper published or to be published regarding IBT's study of any Aroclor product; 14) Any document (a) evidencing or otherwise pertaining to suggestion by Monsanto as to the "results" to be achieved by IBT in connection with the tests by IBT of Monsanto products, and (b) Monsanto's policy in regarding thereto (i) 1965 to 1980 and (ii) currently; and 15) Those documents relating to items a through j first above listed. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, 000017 HARTOLDMON0023240 inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers. financial statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. "Aroclor" shall mean any Monsanto product containing PCBs or their contaminants. "Animal" shall mean any animal, mammal, reptile, bird, or fish used in connection with any study. "PCB" means polychlorinated biphenyl and its contaminants and derivatives. "Toxicology Department" means any department or section responsible for investigating or determining the possible toxicological effects of exposure to products manufactured by Monsanto Company. Respectfully submitted. By: Michael A. Pohl OF COUNSEL: David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Boulevard Allied Bank Tower, 23rd Floor Houston, Texas 77056 (713) 623-8800 000C;;3 HARTOLDMON0023241 REAUD, MORGAN & QUINN 909 Laurel Avenue. Beaumont, Texas 77701 (409) 838-9941 Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania (412) 471-3980 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. CERTIFICATE OF SERVICE I hereby certify that on the ______ day of , 1987 , a true and correct copy of the above Notice of Intent to Take Oral Deposition and Subpoena Duces Tecum was served upon counsel of record either by messenger or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3708 oooc:o HARTOLDMON0023242 Beaumont, Texas 77704 Mr. John M. Johnson Mr.'Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruehwald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 1313 Merchants Bank Building 11 South Meridian Street Indianapolis, Indiana 46204 depntc0 2/txt8 6551 Michael A. Pohl oooc;;o HARTOLDMON0023243 V E X H l B I T HARTOLDMON0023244 TUz VUI: 650 26J0717 <409) 038-9941 EBBS, MM6IN V VBINN LAVTEBS 909 Laurel Beaumont Tezat 77701 Awwr&at: MCI JBLAJKS MCI io: 263 0717 May 13, 1067 Walter Crawford WELLS, PEYTOH, BEARD, ICEBERG, HUNT & CRAVEORD 624 Petroleum Building BEAUMONT, TEXAS 77704-3706 Re: Cedi Scott v. Monsanto Company, B-64-1103-CA Dear Walter: In order to minimize the burden on you of complying with Plaintiffs' requests for item 6 on IBT related documents --- which requests first came December 31, 1966, were renewed via 30(b)(6) notices April 3,1967, and most recently, April 29, 1967 -- I repeat my verbal agreement with you of this morning that item 6 will at this time be satisfied if you provide each summary report regarding the analyses of, investigations of, and other detailed reports on IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products. I understand that IBT may have done as many as 650 separate studies for Monsanto on some 35 products. I also understand that third parties may have done some 330 study validations on IBT studies of about 30 Monsanto products. It is not clear whether the study validations were done before or after the IBT misconduct came to light. At this time, we do not insist on having the 330 study validations. We do, however, want to know the results of such validations, that is, whether or not the IBT studies were or were not found to be valid, and if not, why not. 2 DEPOSITION t EXHIBIT iLeviusKKS'y I uo/z onoc/21 HARTOLDMON0023245 As concerns this particular request reports about IBT misconduct -- I suspect that Monsanto has some summary reports about the tests, the validations, independent investigations, internal IBT investigations, and even governmental investigations or reports. I suggest that you start with the reports presented to the highest level of Monsanto management and work your way downward until you reach the detailed study validations, stopping just short of them. Surely this will simplify your quest, and with the use of the wonderful Monsanto PCB index and document retrieval system and data base, you can gather this handful of documents together in a matter of days --- having already had months since the initial requests to Identify the dangerous documents. Thank you for your ongoing help in this matter. ooor/j-- HARTOLDMON0023246 ix m I HARTOLDMON0023247 Monsanto IWlJlCf , ( <( . TO : <<% , ! ) ^ c/Z ' George Roush, Jr., M. D.__________ September 17, 1975 MONTHLY COMMENTS - MEDICAL DEPARTMENT Corporate Administrative Committee CONFIDENTIAL State and federal agency activities related to PCB's continue to require Medical Department attention to support business group efforts. Recent public hearings in Wisconsin served as a forum for Mr. Schweitzer, of EPA's Toxic Substances Office,to make his pitch again in support of the Toxic Substances Control Act now being considered in Congress. Between now and the end of November, FDA, EPA, and NIOSH will conduct hearings on various aspects of the'PCB's in foods and the environ ment. Questions from outside'Monsanto about the long-term health effects of chemicals used in two Monsanto plants on the workers in the plants have required organization of epidemiological studies. These require acquisition of data on individual work assignments within the plant over long periods of time, collection of death certificates of deceased employees, and comparison of causes of death with frequency of cause to some segment of the general population. We will have to do much more epidemiology in the future although clear cut conclusions are almost impossible. GR/ln ~ DEPOSITION \ EXHIBIT i oo/cl SCM 019722 ooor;^3 HARTOLDMON0023248 xxm I HARTOLDMON0023249 'Monsanto ' |Mt * lOCATlOMl Medical Department A2SA October 13* 1971 AROCLOi^ 1260 te R. E. Kelly M. N. Johnson E. P. Wheeler TO PILE !.< MV IM| I spoke with: Tel: Renate D. Kimbrough, M.D. Pathologist EPA Atlanta Tox. Branch 4770 Buford Highway Chamblee, Georgia 30341 (404) 633-3311, ext. 5218 today regarding her obaervatlon of bladder tumors In rata fed AROCLOR 1260. The observations of Vos and Koenan (Toxicol, and Applied Pharmacol. 17, 656-668, 1970) on polychlorinated biphenyls led them to undertake rat reproduction studies with AROCLOR 1254 (Lot AX38) and 1260 (Lot AK3). The materials were obtained from Glasgow at FDA. Their primary findings ^ have been lea Iona In the liver, and Dr. Kimbrough plana to present a paper on the liver lesions at the 1972 spring meeting of the Society of Toxicology. In addition, ahe haa seen 2 lesions In the bladders of rata fed AROCLOR 1260. The first occurred In a female which died after 6 months on test. Thla waa dlagnoaed as a malignant anaplastic carcinoma of the bladder. The second waa In a male killed after 8 months on teat. The flrat diagnosis waa of epithelial hyperplasia. Sections of both bladders were sent to NCI for diagnosis to Dra. Strauss (?) and Katherine Snell. The diagnosis of carcinoma in the female was confirmed. The lesion In the male waa not resolved. Some pathologists believe it Is a carcinoma, others believe it is -.hyperplasia. Dr. Kimbrough stated that they were trying to analyze the AROCLOR 1260 sample for Impurities, but were having some difficulty with their equipment. I Indicated that we would try to track down material from the sample that they had received. In the event that we cannot locate this lot, we probably can get some material back through Thomas B. Gaines, Supervisory Research Pharmacologist, at the same location as Dr. Kimbrough. If we have an analysis of this lot, we should make the results known to Dr. Kimbrough. Dr. Kimbrough said that they had observed porphyria In some rats with both compounds. She also Indicated that they were contem plating doing a 2 year feeding study with larger numbers of rats to Investigate the problem of bladder tumors. She is per plexed by the bladder tumors, and she Is not emphasizing them in her discussions. However, she has mentioned her findings when PCB's have been discussed at various Interagency meetings. This Is apparrently how Weissburg learned about them. (cont'd.) H DEPOSITION t exhibit K- (r ^"'4-7-1") coir 100Oitt HARTOLDMON0023250 PILE October 13, 1971 Page - 2 - As a final note, Dr. Kimbrough expressed concern over whether PCB's presented an additional hazard to the employees who manufacture It. 1 told here that because of the chloracne and liver hazards, there had been medical supervision of the employees. However, I would raise this point with Dra. Kelly and Johnson for their review. , /blea N / > ) i } 052188 St* ooor;: HARTOLDMON0023251 U iX X ( \ I B I T / I HARTOLDMON0023252 October 28, 1971 Or. Moreno L. Keplinger Induetrial Bio-Test Laboratories, Inc. 1810 Frontage Road Horthbrcnk, Illinois 60062 Bear Kept I spoke with Or. Senate Kimbrough of the E?A in Chamblee, Georgia regarding our proposed visit to her to discuss bladder pathology La rats fed this product. Z Indicated . to her that we expected to have all of the available bladders examined by the latter part of Vovember, and that we would confirm a specific date beforehand. The express purpose of the visit is to exchange Information, l.e., view the slides she has, and let her see any slides that Bon turns up which may be of interest. This was agreeable to her. She suggested that we might bring extra slides for referral to KCI, if we so desired. After Don has finished his evaluation of the bladder sections, and after you and he have had a change to review his schedule, I would appreciate a c-pple'of dates that would be convenient for a trip to Chamblee so that Z may schedule a visit there. "Kindest personal regards. George J. Levlnskas, Ph. 0. Mgr., Product Svaluatloa OJVbks cct Dr. Don Gordon Mr. V. B. Pspegeorge Mr. B. P. Wheeler i DEPOSITION i EXHIBIT j S-M-n o->C SCM 023586 OOOG^G HARTOLDMON0023253 LDXX V I B I T HARTOLDMON0023254 HARTOLDMON0023255 u jx x f i HARTOLDMON0023256 -- Joj**.'*&/ r*-'*\\* ' . * * - ^ , '~u?':U rot tuiorTii Ue Aroelor studies. - _ rwib'Uca.ti.dn w*y\'uV'&fjiOy:rdpci*ilJw< -v ->. ' . in Its nonain^'and; have; ,inatoad provided, descriptive. terns for thooa' lesions not ltovinn;ibo' elnsele oppooirnnea dClivor carclncna,. However. _________________nnJ wish .to llianJt.you Dor the use of your . notorial. Ho iasiona will beldcntiricxlviUi.iyiy "specific coapoiad, -' J.' ' new or in tlu nitu>-o.*^ln fact.'r'mloctud tho'Uldas biiadly to'nprrrent' HARTOLDMON0023257 >': - -SV'fe:M cy^hy^'M ',*" ' r '* ^'^fv V S'i*%' /.^'/>?&'>*r.?T V&.*$vl$;/ :..,..-,-v. ^ : *<, '. -. wV >. 7V ' : ;' '.&$& ' ^ ` ^ 'S, ..`-i- : ' -l - 'O HARTOLDMON0023258 UJXX HARTOLDMON0023259 7.R.Johonnsen - A2SC December 6, 1974 AROCLOR 1254 DC t 1 i: 74' E. P. Wheeler - A2SA W. B. Papageorge - E2CK 0. Roush A2SA /o In a recent lphone conversation between Dr. R. Kimbrough (CDO) and 0. J. Lcvinslcas, Dr. Kimbrough expressed the telief that rICI had recently contracted out a carcinogenicity study involving AROCLOR 1254. Consequently, I made a call to Dr. Sidney Siegel at IICI. Dr. Siegel indicated that he had no knowledge of an outside contract with ARCCLCR 1254 and further stated he doubted there was one. He did tell me that AROCLOR 1254 has been under test at IICI in their bicassa.y operation program since September 1972. He reported that the experimental part of the study load "been completed aj^of October, 1974. His expectations are that the necessary histopatholosyrriirbe completed within the next * 6 months, at which time a meeting could be set up for discussion or Tile test results. '----------------------- . - T7 A quick check of the April 3* 1974 list of chemicals being tested . under the IICI carcinogenesis program did not reveal that AROCLOR 12^4 was on test. However, the NCI list does show C02664 BIPHENYL, CHLORO as part of the program. If this is the study he is referring to, it could explain why we had not picked it up sooner. / (m) fhft/ Ct. o /bkp / Frederick R. Johannsen ,V Ol* ++ vj * ** - >jL. y-Jt; tfaM^ (j/2 ,o/i - deposition f EXHIBIT 00/ osifc*8 SC* OOOCoO HARTOLDMON0023260 UJXX I \ B 1 T n HARTOLDMON0023261 ) OCFARTMCNT OF HKALTH, KOUCATIOH,>NO WKLFARt PUBLIC HCALTH StftVICC CtMTU rod OIIUU UMTML ATVAXTA, QKOMOiA MU1 rn-i* n< tttm uuu Decanter 6, 1974 !7 i Dr. Georgs ierineha* Monsanto Industrial Chrsdcals Conpaay 900 I. Lindbergh Boulevard SC. Louie, Missouri 63164 Deer Georgei 1 sa enclosing brief outline of the experiment Aroelor 12-72-A . for your information. A copy of Bob Squire* letter 1* elao todoied.' Sincerely yours, . Enclosure Senate D. Klsfcrough, M.D. Toxicology Branch HARTOLDMON0023262 xxm I I \ \ I B I T HARTOLDMON0023263 izms/r -. .&/. &r:. vf~:.i3sp '.. F'^ ,, ' ""Jfexfs^fe: . AP. e'* ft..;. 3 Dr. Donovan "Oordon;____ ____ , .. Industrial B20-TEST libdnioneS^fi 1810 Frontage Hoad ttorthbx'ook, Illinois vfiSSfei&ir* __^r* a'lr.r**' -*? . *vi\ - - r * ?r ' ' Dear Don As a follow-up to our phone conversation yesterday, X aa . , sending you copies ' of.Ithe Information on Arodor 1260 study - . which was sent to ne by Dr* Kimbrough.. You nay find this ` lnfornatlve In preparing.for your subsequent'dlscassloo VV'-f When you have had a' chance, to review yourschedule, would you . kindly let ae know What days my be convenient for a nesting. -' This probably will be In the Washington area and Z anticipate j that you, Renate Kimbrough, Bob Squires and I will attend. . . * 'jLJ:-`-S< Vv;WV-M.a "i^f Sv^: v;" . < :* ^Ejpiv 000032 JflJ HARTOLDMON0023264 SOI 70* fmiCAIIOH o* CI1CTJIAT10H Aroclor 12-72-A Animals Random brad female Sherman aCrala rata raarad uadar specific pathogen-frae conditions vara obtained froa tha SCSC farm at Lswrencavllla, Ga. Aroclor 1260 Aroclor 1260 (Lot number AX-3) vaa supplied bp Monsanto Industrial Chaalcala Co., St. Louis, Mo. iI Statistics Tha students t-tast vaa usad for comparing body valghts and weight gain. Methods . Pour hundred 21-26 dap-old veanllng female rats, weighing 46-97 g, vara distributed into 2 groups of 200 animals each according to a table of random numbers. Each animal, was valghad and given an Individual ear 1 tag number. Tha animals vara housed 10 rats par cage, food and water vera provided ad libitum. Two hundred animals wera fed tha control diet of ground Purina laboratory chow; tha taat animals wera fed the saaa diet fortified with 100 ppm Aroclor 1260. Aroclor 1260 was Incorporated Into tha diet bp dissolving 5.2 g In ethpl ether than adding this to 100 g corn starch and allowing tha ether to evaporate. Tha PC3-cornstaxch mixture was blended with 345 g ground ehov, than pulverised In a mortar and diluted with additional ground chow in a bakars mixer to give a 1000 ppm concentrate. Tha concentrate was further diluted to obtain 50 kg of 100 ppm diet. Tha diet was prepared every 10-14 daps. Random samples from tha final mixes along with samples of control chow vara taken at regular intervals to determine PCS levels la tha control chow and to verlfp tha 100 ppm fortification level. Ground laboratory chow was also checked at intervals for aflotoxln at tha dls trice of flea of ehe Food and Drug Administration in Rev Orleans. Tha combined body valght of rats In each cage vaa recorded weekly until tha rata vara 6 months old, bi-veeklp until 12 aontha old, and monthly thereafter. Individual velghta vara recorded only sc tha onset of tha experiment end at death or sacrifice. Tha animals vers observed SCM 022811 nnnr.PA HARTOLDMON0023265 I Axoclor 12-72-1 - Page 2 briefly each day tad animals exhibiting debilitating signs or latte tumors ware removed from the group cap tad housed individually. At aech valghlng tha animals vara axaainad Individually and abnormalltla's notad. food conjunction van eaasurad on all rati during the firat two weeks of tha experiment, than during vaaka 3, 8, 11 end 20 and tvary 12 veeke thereafter, Antopeiee vara performed on all rata that died, lata that were atcrlficad vara billed under ether anaathaaia by aeverlng tha vena cava. Tiaauea vara fixed in buffered forealin and stained with hematoxylin and eoeln. Additional special stains vill be conducted. Xasulta During the course of tha study 4 animala in tha teat group and 8 controla vara sacrificed prior to the final kill. Theaa aniaals had either large tuaora which had ulcerated or hindered movement, or tha animal appeared moribund. In ell. 14 animala la tha teat group died; 17 animala in the control group died. One animal.vaa accidently killed in aech group early in tha study. Ifo definite doee related signs of toxicity vara observed In the teat animala throughout the study. Comparative curves of body weight gain and food consumption on the basis of body might, as mil as PCI intaka of tha tsst group are given in Figure 1. 1 alight decline in the rate of weight gala of the teat group cohered to the control group began about 3 months after onset of tha experiment. Mean final body valghta vara 420 g (S.D. 72, S.Z. 3.4) for tha control group and 392 g (S.D. 62, S.Z. 4.6) in the teat gToup; the difference vam statistically significant (p<0.001). Pood consumption (gm/rst/dey) vaa cooparable in both groups throughout tha study. Kean might gain van 330 g (S.D. 70, S.Z. 3.3) and 323 g (S.D. 60, S.Z. 4.3) for tho control and teat .groups, raapactivaly; tha difference in might gain vaa alao statistically significant (p<0.001). ?CB intake declined from 11.6 mg/kg/day during tha first week of exposure to 6.1 mg/kg/day at 3 months of exposure and to 4.3 mgAg/dsy at 20 months. A 6 g mean might lose occurred in both control end teat groups during tha 6 make prior to the final kill. 1 SCM 0228U oooc HARTOLDMON0023266 Preliminary of Microscopic findings (liabrou*h) Please Hoc*: Sections of All Tissues Listed Vers Rot Aval lib Is for All Animals Timor Type Controls (Tissue of one rat autolysed) Experimental Rees Maanary Gland: fibroadenoma 19 14 Adenocarcinoma 7 0 Pituitary: Chromophobe Adenoma 38 30 Csreinoas Pituitary 0 1 Uterus: Endometrial Polyp 20 23 - Endometrial Sarcoma 2 3 and 1 adenocarcinoma Thyroid: Medullary Call Tuaors (parafollicular) . 26 18 I Liver: Modular Hyperplasis 3 (8188,8192,8232) 7 . Nodular Hyperplasia and Hepatoaes 1 (8231) 134 Nodular Hyperplasia* Hepatomas, and Hepatocellu lar carcinomas 0 Adeaoflbroels 0 14 4 Bladder: Papilloma 1 (8317) 0 Is addition, a nusfaer of other occasional tuaors were found. SCM 02281 000CC5 HARTOLDMON0023267 Preliminary Liat of Microscopic Pladlnga - Paga 2 Out of this group t 4 control rata and 4 rata la tha axparlaaatal group vara aacrlflead aarly because they vara tick* All tuaora of thaaa aalaala, outalda of tha liver laalona, ara laeludad la tha praeaedlag 11at. Tha liver laaloaa naad to ba addad. Tha aalaala ara aa follow i Controla . Experimental 8194 8131 8121 8234 8331 8347 8413 8317 SCM 02281b HARTOLDMON0023268 Figure 1 BODY WEIGHT Months i Ii I 1 l \ HARTOLDMON0023269 ecuosjo Figure 2 FOOD INTAKE AND PCB CONSUMPTION <> oo rr\\jj o Xjin. 1i * i i l j w HARTOLDMON0023270 LUXX ! II ! ! I \ I B I T # ( i I HARTOLDMONOQ23271 AROCLCR 1260: Meeting *t KX# January 31# 1975 Ti*e purpose of this meeting was to review sections of liver tissue from Or. Kimbrough's 2-year study in which feaale rats were fed 100 ppa of AROCLOR 1260. Drs. Kimbrough end Squire had studied and classified the findings. Or. Levitt, either a visiting fellow or a post doctoral trainee did not participate to any great extent. He was aentlooed as having a good knowledge of liver. Dr. Rlcnter had viewed the slides froa our 2-year study in which rats cf both sexes had been fed 100 ppa, 10 ppa or 1 ppa of AROCLOR 1260. Subsequently, be and Or. Cordon reviewed sections of liver froa all aniaala In this study. Three conclusions can be drawn. 1. In our earlier study# the severity of liver lesions wan greater in fearles than in salas. ?. To a large extent, substantially the saae type of leslonr were observed in both studies except that- the lenlons seemed to be wore advanced in flab rough's study. Although the re was tone variation in terminology, the findings were reasonably close. 3. There were definite liver adenocarcinomas in Kimbrough's study. Or. Richter expressed the view later that 2 anlsals in our study approached the type of lesion brough had observed, but there was agreement by Drs. Cor don and Ric.iter that Or. Kimbrough's rats had developed a lesion which they had not observed In our earlier study with AROCLOR 1260. If. Drs. Cordon or Richter feel that I have not suamarixed this seating correctly, or if they desire to saend or expand ay remarks, I invite them to let m know. % /bkp 2/3/75 -George J. Levinskas 0^0033 HARTOLDMON0023272 U IX Z I B HARTOLDMON0023273 S' -" - -r'-tlntaa fit* Dr;?Kimbrough* ^.rv ^rats wara fedlOO ;ppmV ' ^8-jrear? MSCtAu- dy 3a. \ j?.'Vj'a0^a3yS!^s^*!VAi>fc ^'t.;^XT..-t'C' findings.; _____________ Dr.^Ivitti;-'eit2*ar,Su:,vialiing fallow ok>t thS^ - `i- ^doctoral tralneedid ncrtpaxticipnte.'to any great " v~-.- Fa?7* . v**v.. v -'v-. . *--: 'a <jK<V\xTsf* . .. . To a large extent ^ Substantially the saae typeof lesion* were observed In both studies exceptvt)at-'.the Jjesions ^ ,.. seemed to be more advancedln Kimbrough'S study.7* Although tbon ............................. ` ---- were 3. There were deflnltefllver..adenocarcinomas in Xlnbieugh's .. . study. Dr. Richter .expressed tbe view.later, that a sm-;-. ^V' . sals In our study1 approached the gtypeiof lesion ' - ... trough had observed, but there was agreeaent by;Dr*. ;0or- ,J%, > -..1 don and Rleater that Dr.- Kimbrough's rat* had .developed `tTK- *a lesion which'they, had not'.observed'In our. earlier study '' meeting correctly, or. If they desire .to anend oraxpanday vv T 4olSa ttiea +s* 1*0 m Vn nsr ssTaf'' '*-- xlJ HARTOLDMON0023274 UJ X X HARTOLDMON0023275 K TOXICQlOaT (NVIHQNMCNTAi tCItNCKS CMCMiiriT AUNT CIINCCI MKOieAk seiKMcta !}tukAtf/u&L BIO -TEST JtcrfytfjObbyuAb, JJnc. 1810 S*ONTAG8 *OAO NORTH8ROOK. ILLINOIS 60062 March 24, 1975 ** COOK Jil rCLCMONC I7MOJO George J. Levinskas, Ph. D. Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 63166 Dear Or. Levinskas: . I am enclosing the trip reports prepared by Or. Richter and myself regarding our meeting at N. C. I. on January 31, 1975 with Ors. Kimbrough and Squire on Aroclor 1260 (IBT No. 622-07298). . .. _, .. ' . . *.t The micro-slides of the liver sections from all these animals and' those from Aroclor 1242 and 1254 will be forwarded today, under separate cover. We are also mailing the reports relevant to these studies today. The charges for this additional work will be billed under a new Number (IBT No. 641-06672) instead of the previous number. Sincerely O. . Gordon, D. V. M., Ph. D Section Head, Pathology Dept. DEG:hb Enclosure DEPOSITION } EXHIBIT | /S'! SCM 023632 HARTOLDMON0023276 February 2, 1975 Trip Report On January 31, 1975; Ora. Richter, Levinskas and myaelf met with Drs. Squire and Kimbrough at the National Cancer Inetitute in Bethaeda, Maryland. The purpoae of thia meeting wee to review the slides and data from a 23 month oral feeding atudy with Aroclor 1260 in rata that waa con ducted by Or. Kimbrough while at the E. P.A. In her atudy, 200 female rata (Sherman atrain) were fed 100 ppm of Aroclor 1260 in the diet for 21 months, while 200 female rate of the same atrain served as controls and were fad Purina Rat Chow. The Sherman Rat is a random-bred animal . ' .. ` ,. ***.* . derived from the Osborn Mendel Strain. The animals were 21-26 days of ' age when the atudy began, and were sacrificed after 23 months on the study. Therefore, there was a 2 month recovery period at the end of the atudy in which the teat material waa removed from the diet. The details of the ex perimental design, procedures and results that were forwarded to Or. Lav- inskaa are attached. In summary. Dr. Kimbrough found a rather high inci dence of hyperplastic (nodular hyperplasia) and neoplastic (hepatomas, car cinomas) lesions in the liver of the test animals. The slides were subsequently reviewed by Dr. Robert Squire, Head of the'Tumor Pathology Brlnch of The National Cancer Institute, who concurred with her findings, although, the ' . terminology he used in classification of the lesions varied slightly from Dr. Kimbrough's. In this regard, it was his impression that all hyperplastic nodules should be regarded as "pre-cancerous lesions" and the term "hepatomas" SCH 023633 000CI3 HARTOLDMON0023277 Trip Report 2 bo deleted and re-classified a* carcinomas or naoplaatic nodulaa. Tbia classification system waa baaad on a Liver Tumor Workshop that was held on Dec. 11-13, 1974 in Silver Springs, Maryland which waa attended by a small group of pathologists from the regulatory agencies, research labora tories and Industry. In view of Dr. Kimbrough's findings, additional sections of liver from a 2-Tear Oral Feeding Study With Aroclor 1260,. in rats, conducted at Indus trial Bio-Test were processed and evaluated. In this study, there was a 3, 8 and 14 month interim sacrifice which enabled one to determine the ap proximate time of onset of the liver changes. No interim sacrifices were, conducted in Dr. Kimbrough's study. The additional flides prepared at . . Bio-Test were examined by Dr. Richter and a tabulation of the liver findings are attached. These slides were reviewed by Drs. Kimbrough and Squire at our meeting. Although there was some disagreement on the terminology used in classification of the lesions, it was apparent that the incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) was greater in Dr. Kimbrough's study when compared with the results of the Bio-Test Study, even though the same level of test material was administered in the diet. There are two possible explanations for this difference: 1.) The strain of rat. When questioned. Dr. Kimbrough had no informa tion on the spontaneous incidence of hyperplastic or neoplastic liver . lesions in the Sherman strain. However, the incidence of spontaneous liver cell hyperplasia would appear to be extremely low since this finding was reported in only 4 controls. The incidence of liver hyperplasia among control rats of the Bio-Test study was also very low. The Incidence of spontaneous hyperplastic liver nodules reached 20f* in the Fischer Rat 000041 HARTOLDMON0023278 Trip Report 3 and In loma of the other strains. 2.) Sex difference* On the basis of our atudiee and thoae of other in ' read gator a in rata and mice, it waa found that the incidence of hyper* plaatic and neopiaatic liver lealone are much higher in femalee. The reaulta of the liver tumor workahop are to be publiahed in the near future and will include the participante* The viewa of thia group, regarding claaaificatioa of hyperplaatic and neopiaatic liver laaiona is based on the pathogeneaia of auch leaione in rats that have been Induced by known carcinogens auch as the nitroaoaminea and 2. - acetylagdnoflorine (2-AAF). The adaption of this terminology and claaaification ayetem by the scientific community will have wideapread implicationa regarding aaaeaament of aafety of chemicals and drugs currently on teat and thoae previously studied. Attached are Dr. Richter's comments in regard to Dr. Kimbrough's findings. . Donovan E. Gordon, D.V.M. ,Ph. D. DlpL., Am. College Vet. Path. HARTOLDMON0023279 Or. Donovan Gordon Indue trial Bio-Teat Laboratories February 3, 1975 I wish to summarize my observations of the lesions seen in Or. Kimbrough's study of Aroclor 1260 and the comparison of diagnoses with Or. Kimbrough and Dr. Squire. 1. ) The criteria that I used for diagnosis of the lesions are similar to what they used. 2.) My evaluation tends to be a little more conservative than theirs. For example: they ^tould call some of my hepatomas careinoma but with some question. 3.) Therefore, if we both read the same slides there might be a little variation in numbers of lesions in the different categories but no major difference. 4.) Dr. Squire and Or. Kimbrough are using a new and revised terminology for the categories and list them as follows: My Terminology (Classical Use) Dr. Squire's (Revised Terminology) Focal Hypertrophy Cellular Alteration Nodular Hyperplasia! Hepatoma J Nodular Neoplasia or Neoplastic Nodule Carcinoma Carcinoma 5.) However, the lesions in Or. Kimbrough's study were more severe those in the Bio-Test study. The lesions that she and Or. Squire are calling carcinoma are also carcinomas by my criteria. I would HARTOLDMON0023280 conclude from in exunimtioa of their miteriil thit Or. Kimbrough*a tudy demonetrited cireinogenieity. Ward R. Richter, D. V. M. Dipl. Am. College Vet. Pith. SCM 023637 0000-17 HARTOLDMON0023281 u ixx I B I T f HARTOLDMON0023282 KJ 25, 1970 Otis E. Fancher, Ph.S. Director Industrial Bio-Teat Laboratories, Inc. - 1810 Prontage Road Northbrook, Illinois 60062 Osar Otis: * This 1st tar will authoriss you to procssd with a study with tha leghorn chicksns with ths Aroclor 1242, lot numbsr AL-55* This should dupliests ths study you did previously with Aroclor 1242, lot nuabsr AX-255, except in this instance you haws suggested using dietary lave la of 2, 4, and 8 ppa. This sample of Aroclor represents our current regu lar production which involves some clean-up insti tuted since the previous sample was aade available ' to you. Ve would hope that we might find a higher "no effect" level with this sample as compared to the previous work. - As soon as suitable reference standards are available, the research laboratory is St. Louis will more care fully characterize the two samples in terms of possi ble minute amounts of contaminants. Best personal regards. Sincerely, Elmer P. Vheeler Manager, Environmental Health SCM 023565 o^or;:ri HARTOLDMON0023283 U iX X I \ HARTOLDMONOQ23284 TO*iCOLOV KNV1IIONMCNTAL SCii CMCMISTWV *-** SClCNCCS McMCAi acicMeca fCB Jn&uMai BIO - TEST J!abo*aiMie&, Sno. - 1B10 ^RONfAGH ROAD NORTHBROOK. ILLINOIS 60042 anca eooc SI* mimoHC iffi< September 30, 1971 Dr. George J. Levinskas Manager, Product Evaluation Monaanto Company 800 S. Lindbergh Boulevard St. Louie, Mieaouri 63166 Dear George: Endosed are the. correction pages for the three teratology etudiea with Arodor 1242, Arodor 1254 and Arodor 1260. I die cue *ed die interpretation of die renal caudal ectopia, found with Arodor 1254, with Dr. Keplinger. He ie in agreement with the interpretation that thie ie probably not a specific teratogenic effect but a general indication of the toadcity of the materiaL hi reviewing the reeulte from the three generation study there appears to be a reduction in both 5 day and 21 day pup survival in die group fed die highest level (100 ppm) of Arodor 1254. Thie was not observed in the other Aroder groups or in groups fed 1 or 10 ppm, again suggesting that die level of Arodor had a general toxic effect. As I pointed out in our phone conversation, die small differences observed la sex ratios are not significant. A statement to that effect has been inserted. If you have further questions, contact me at any time. PLW :1am Enclosures Sincerely yours, Paul L. Wright Section Head, Toxicdogy r- deposition f EXHIBIT \SLEV/A)S#S-n\ cO<. QO0C '0 SCM 02350^ HARTOLDMON0023285 xxm i \ l B I T / I HARTOLDMON0023286 April 28, 1979 * deposition I j exhibit .3 $-) oj/cj Dr. Otis Fancher 624 H. Abrego Drive Oreen TSlley, Arizona 85614 . Dear Otisi Z an aahaaed that Z have fceenlso slow; it* reviewisg the aanuscripta and sending you our eosawnta. Z guess Xsp's sending ae the chronic rat and dominant lethal and nhlcken study on the 18th of April finally atlx fed as to boys. First of all, Z agree we should hold up the publi cation of the chicken work until the current study is eoapleted* .* * - the only other eoonent I hare of substance la that Z wish we could vo~k la several paragraphs reviewing the Troon work on vapors published la 1956 (see en closed xerox copy of reprint), k logical, place for a review of those data would be at the top of the ~ 3rd pegs of your review paper, before your paragraph at the top which begins with *Durlng succeeding years llttla Information was reported eoncesnlag . the toxlohlpgy of taeJC3j3r>til goLaugh! la etal, 1963. ete.- . goat of the review articles on the FCB'e have failed to indicate awareness of the Treoa work* Z believe this is because the Rlsebroughs. Feakalls, etc*, ete* -- the nviromaontalists -- have not encountered the Anai'lcan Znduatrlal Hygiene Association Quarterly la the review of the literature nor .la It likely that . they ever heard of the AXHA, .. .- Zn any case,' Z think the work la worthy of review even though it waa inhalation data particularly since Drinker's earlier work in *37 1* frequently nentlooed and. this, again concerned Inhalation* ' . v ' i`-% . * . *- . e. .1.-. *. Zn the last paragraph of thla sane review paper you . "*. vi` r-SCH 023539 HARTOLDMON0023287 Dr, Otis Fancher April 28, 1972 Pagt - 2 - refer to the perniaeable aablent concentrations of PCB*l" an auggeated by Xlrlnicer In 1939. Aotually the AC0I3 established the Threshold Halt valuta baatd oo Trton'a work rathtr than the tarlltr Drinker work. Z btlltvt wt aay havt arrived at our high dlttary level of 100 ppu in tht atudlta dont at Bio-Teat baatd on tht Treoo work rathtr than Drlnkar'a. George Ztvlnakaaa la reviewing taeh of tht paper* ' to sat If you. Bap and I havt Biased typo'a and ha ay wall suggest soot graanatlcal change# igjoh wt will tntar on our eopy and atnd ztroz*a to you and Bap. . Z hopt to gat ooplta of all of tht atudlta in tht hands of Bill Fapageorge, Scott Tucker and tht lawyers next wttk. Z do not antlelpatt a lot of ohangta frou thouand hopt that tht attomtya agrtt that wt oan go ahead with publication. Boat personal regards. Sincerely. m/bka cos M. L. BepUnger Xlatr P. Wheeler Mgr., Environmental Health Medical Department oom . SCM 023590 .* HARTOLDMON0023288 V e: x H I Bi I! t; ( i HARTOLDMON0023289 * / HnduMd, BI O-TEST Jldyyuzt&ues. Snc. 1810 PRONTA3C AC AO NOATHBBOOK. ILLINOIS 80082 OXICOkOdT CHVIAOMM C*TAV tOCNCCt CMCMltTIr ' ^u>r scicmccs MCOtCAV BCiCNCCJ April 18, 1975 ArR 21 *<* 30C JU t(lx#monc j;mcjc Dr, George Roush, Jr. Mon seats Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear George: - 1 fully appreciate that the meeting an PCB's today was not completely satisfactory and that many nagging Questions remain. The enclosed is a brief summary of my personal views and I would appreciate any open and frank comments that you all may have. Please let me know of any action that you contemplate in the way of seeking additional assistance in pathology or in contacting federal agencies. We will be pleased to be of help in any way that you may wish. It is my feeling that we need to get together again within the nest few weeks to continue our discussions. * Very truly yours. JCC:AR J. C. Calandra President cc -Dr. George Levinskas Mr. Elmer Wheeler Mr. William Papageorge SCM 022745 deposition EXHIBIT IjlEVUftgfrS- \[ 11 *s -wn 03 `QCSC HARTOLDMON0023290 xxm I B I T HARTOLDMON0023291 a '*..: - *;_ . IV' . Dr. Gaorga .":-.v\**:r*-V, -* 190 H*. 'UntborghBl St.; toat<Sw~t?'tlfUe29rUM4n*j*w*:!i & Or* . KSS-;.!?' 'Q: yj#* . ,j-i--d---t-i-w. .-at: U. - m7, <tan ity jjaMiaamlaaS additional '. ~ Til IttpfB Jmli.,*:., ' a11BT Ho.~f2.ftTlTha iaitio*''wm Sit from lb* original block of nt>jd4 'Uiiw^;.Jkt^Vq|(t|,C|>|tilo* vaa mI|IUt cUaaiCod try Dr." ftkiUf il l hopetoco*. ~^Jpoa rarlaw,7P>. 'Richter h eUiilflti thla lotion owhlu kyptijptttU o*d I ooocmr. Howovar, yon will ilio note that whan tUttoMl'McliaM of llm from this animal worn pntiiHd (ST Study Mwrabay41-66&7). no Urtr aliaraliana wara ' *'< which atteatatetfarJocal hater* of than*laatoaa.- 'i - > /''<* v v"= -J`r- *.~...T....W.. /Thla eiuct,J raport of IBT No^:< 'i#ak>o.V J`' dictating i sllda of tha abtit A,' r>r.'/ . . `.I _ a &!> Aa.z" \i >r. -^VC r-*4 " a E. Gordo*. D. V. M.. Ph. D. . ^: . , Saction lluf, Pathology Obpartmoal aaclonrt. | i ooor;c.3 HARTOLDMON0023292 1 > j 1 1 1 1 1 1 ' J :::::: I ; C ontrol +1>* cn EA K O 0 31 *- 0 C-OI lOOppm r? ^: IC : s :: J ::::;: j a I? : i j ;: j< > J? S se 2. * 8.? +X t x+ x * t f Cytoplasmic vacuolatlon of hapalocytaa Focal nacroala of hapalocytaa Focal lymphoid Infiltration* j j Focal byportrophy of hapalocytaa a <o<d u >> D *d onn *d Nodular hyporplaala of bapaiocytoa Focal blla duct kyporptaalo Hepatoma c a H rm eS' *0 Clislanglo-hepatoma (letlculum Coll Sarcoma ma la atallc Mammary tumor, metastatic T** 1 ||I|M'I'I **!.. fM HARTOLDMON0023293 u ixx I B I T # HARTOLDMON0023294 s Monsanto MoflltntS Comgiity 800 N. Lindbtrgft louUvird St. louii. Umouri 83108 Fton: '314! 194-1000 July 18, 1975 Dr. J.C. Calandra Industrial BIO-TEST Laboratories 1310 Frontage Rd. Northbrook, 111. 60062 re: AROCLOR 2-year Rat Feeding Studies Dear Joe: The attached table summarizes a comparison of the 3 revised. AROCLOR reports (12^2, 125^, 1250). * In 2 instances, the previous conclusion of "slightly tumorigenic" was changed to "does not appear to be carcinogenic". The latter phrase Is preferable. May we request that the AROCLOR 125report be amended to say "does net appear to be carcinogenic'. The number of hepatomas reported for AROCLORS 1260 and 12^2 have been Interchanged. This appears to have arisen from con fusion regarding the numbering of the animals. The original reports show tumors in animals with numbers in the 100-300 range for AROCLOR 1260 and in the 500 to 800 range for AROCLOR 12^2. This leads me to conclude that the numbering scheme shown in the second set of reports is correct. With AROCLOR 125^ confusion is compounded. The original report showed tumors in animals with numbers in the units to teens, but the revised report shows animal numbers ranging from 40 to 1000. Can this be straightened out? I was unable to reconcile the differences in the animal numbers between the first supplemental report and the original reports, I had inquired as to the changes in the numbers. As I recall, I was told that the sections had been renumbered when the new slides were made and that a key relating to the sets of numbers received jui Z1 l-o HARTOLDMON0023295 Dr. J.C. Calandra July 18, 1975 Page - 2 - would be supplied. This has not been done. It may not be necessary for AROCLORS 1260 and 1242, but AROCLOR 1254 remains unresolved. Insofar as I can see, the remainder of the reports appear acceptable. Kindest personal regards, Sincerely, George J. Levlnskas, PhD Mgr., Environmental Assessment and Toxicology /bkp att. cc: Dr. George Roush, Jr., M.D. G^GCCC r oA A * m HARTOLDMONOQ23296 Product Supplemental Report #1 (mailed) AROCLOR 1260 . conclusion hepatomas. , . slightly tumorigenic t* 3 range of p. 9 p. 10 p. 11 p. 12 P. 13 p. 14 test animal nos: 600 to 800 1000 series 70 to 100 500 to 600 600 to 700 700 series Supplemental Report #2 (JCJ delivered) does not appear carcinogenic 7 100 to 300 800 to 900 lOttO 40 80 to 200 200 to 300 200 to 300 AROCLOR 1254 conclusion hepatomas slightly tumorigenic 6 slightly tumorigenic 6 . AROCLOR 1242 conclusion slightly tumorigenic does not appear carcinogenic hepatomas range of test animal noa. 73 as in report #2 as in report #1 for AROCLOR 1260 AROCLOR 1260 SCM 02280^ o^oc HARTOLDMON0023297 xxm i I B I T # l I ! i Ii i i HARTOLDMON0023298 /]/icei rouicoco^y (nvhonmimtav *ci CM CM' *T*V LAMT ICKNCfl MCSICA4 ICKNCfl Jrt&uA&iuil BIO -TEST J!&6<MCiX&ue&. Sttc. 1810 FRONTAGE ROAO NORTHBROOK. ILLINOIS 80062 August 4, 1975 ACA COOC Jit rtiONONf in*jou Dr. George J. Levinskas, Manager Environmental Assessment and Toxicology Monsanto Company 800 North Lindbergh Boulevard St. Lotus, Missouri 63166 Dear George: \ -' Re: Aroclor - 2 Year Rat Studies : " in regard to the comments and questions covered in your letter dated July 18, 1975, pertaining to the above, please note the following: 1. We will amend our statement in the last paragraph on. ' page 2 of the Aroclor 1254 report to read, "does not appear to be carcinogenic" in place of "slightly tumorigenic" as requested. 2. In regard to the animal numbers in the Aroclor 1242 and 1260 reports, they are correct in our final revised report. In the original reports, the Aroclor titles for these two materials were reversed. 3. The animal identification numbers appearing in the reports on evaluation of additional liver sections are the same as those in our original report. The animals were not renumbered. 4. We cannot find any discrepancy in animal identification numbers in the reports (original, re-evaluation, final revision) on Aroclor 1254. However, in the report on re-evaluation of additional liver sections dated March 24, 1975, there was a typo graphical error on page 1 which referred to Aroclor 1260 instead of 1254. Perhaps this is the basis of your confusion. I hope that this will serve to further clarify the situation. Thank you for your assistance and cooperation. JCC:AR Sincerely yours, J4*- J. C. Calandra President POSITION" 1, r EXHW' [I L* itia-j SCM 023620 G0QC.G3 HARTOLDMON0023299 UJXX I v I I HARTOLDMON0023300 *.w-W tlO-TIST 16 U. Summary la Dott iMtiBUt, the spectrum of tnilwitf'nUlid histopathologlcal findlaff la the liver from thie rctviluattoa did nod differ significantly from that previously reported la our original report doled November 12. 1971. There were ela hepatomas detected among six of the onioiols at the highest treataeat level (100 ppm) of the 24-Mouth Sacrifice. No hepatocellular '* " * .. . - \r .. . . . reported ar/Trfgsrded'ss negeas .. - - . - - -t- .I;'-1 *" \ J**** .1 h}"p<rpla*tic in nitur'* and tfcvy or# morphologic ,e * t" ' oi an tcapd'-# response of the Uver ascribed to blotrans/ormation of the test material. The latter lestons were confined primarily to teat animals of the 12 and 24 month v.. . sacrifices and they were dose-related la incidence aad severity. , >i: * '....... * * *" ' :r* y;`-^****w*i-* Aft-w<,". .. . k - v In conclusion, Xroclor U54 do*s not appear to b carcinogenic in rats ... ,, . .. v * ''&T'* ^ '* . J fed lor two years at' levels up to and Including 100 ppm. \ Respectfully submitted. ^ * : : -** ;'/**-uK . INDUSTRIAL 310-TEST LABORATORIES. INC. '#c *1> . HARTOLDMON0023301 rc x rti I V I I III HARTOLDMON0023302 UrtduiUuil BIO -TIST JahoMittHieA. Ate. V. 1*10 FRONTAOe ROAD NORTMiROOK, ILLINOIS MM3 /' August 5. 1975 Georg* J. Levinskas, Ph.D. Moniinto Company 804 N. Lindbergh Blvd. St. Louis. Missouri 83165 ` Dear Mr. Levinskas; . Res 1BT No. 641-06672 - Hlstopathological Evaluation at Additional Liver Sections from Rata ot a Two - Year Chronic Oral Toxicity Study at Arcelor 1254 Enclosed pieAce find 3 copies ol page 2 from our revised laboratory report dated March 24, 1975, prepared in connection with the above study. Very truly yours, . . ^>3 ^leu/uun ' J. C. Calandra ' President JCC/fd t DEPOSITION J EXHIBIT ^LcdivseAV <34 f.y 1 t HARTOLDMON0023303 UJXX I I I B I T HARTOLDMON0023304 August 14, 1975 Dr, J.C. Calandre Industrial BZ0-TS3T Laboratoriae 1810 Frontage Road ^ Sorthbrook, HI, 60062 rat AROCLOR 2-year Bat Feeding Studies Dear Joe: With respect to your latter of August 4, 1975* we appear to be reading froa different scripts, . The attached copies of pages froa the revised AAOCLCR 1254 report (ZBT Bo. 641-06672), dated March 24, 1975 shove that virtually all anlaal numbers have 3-digita. Bte exceptions include a'4-digit number on page 10, a series of 2-digit numbers ranging froa 71*70 on page 11, and slides started Set or Zx'appearlng on pages 12 and l4. Copies of pages 83-88 froa the original report, Z3T Jib. B7298 dated Bfoveaber 12, 1971 also are attached. These contain the tables listing tumors, and they are the only ones in which individual animal numbers appear, Ae summarised below, there is a repetition of low digit numbers in each group. Male ret nos. Female ret nos. Control 1,2 W-Wf50, AHMLfiM I2b4 1 1 ppm 10 ppm 100 ppm 1*2 3-13 1.2 3-H 4,5.7 1-3.6* 8-15 The only pages which contain similar numbers are page 11 of the revised report and page 83 of the original report. How- 2 DEPOSITION 1 i EXHIBIT 1 ^>1-21 U*C I SC" 22S07 oooc.ci HARTOLDMON0023305 Dr. J.C. Calandm August 14, 1975 Page - 2 - aver, even in those Instances where the saaa numbers appear, there are inconsistencies as shown below1 animal no. l 46 47 4e 50 52 12 66 ifroup and sew , JSSSJH 1 ppa Female 1 ppa Female 1 ppa Female 1 ppa Female 10 ppa Male 10 ppa Female 100 ppa Male 100 ppa Female all shown as control females Although "it is of minimal significants, pages 84 and 85 of the original report are aisnumbered in that page 64 is a continuation of the table which begins on page 85. The page for the AROCLOR 1254 containing the phrase "does not appear_to be carcinogenic" has been received, Ve note the interchange of the numbers 1242 and 1260 on those 2 AROCLOR reporta, and will destroy the mislabeled copies. Sincerely, /bkp att. George J. Levinskas, IfcD Kgr., Xnvironaenta 1 Assessment and Toxicology 02zaoa sc* 000CG2 HARTOLDMON0023306 E X H I B I T # f HARTOLDMON0023307 Toneouoar ' CNVlAONM CMTAk ICIfNCtl CMCMIBTKT plant sciences . MCO'CAL SCICNCCS 9M&jbt*tAjjal BIO - TEST JPd&uiiyu&. 9nc. 1810 PRONTAG8 ROAO NORTHBROOK. ILLINOIS 60062 October 17, 1975 Fftf f'M *** cee* nt mifMSNC 171-lOJO George J, Levinskas, Ph. D. Mgr., Environmental Asaesament fc Toxicology Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 63166 Dear Dr. Levinskas: I am returning the black and white photomicrographs of Aroclor lesions in the rat liver that were taken by Dr. Pour at the Eppley Institute in Omaha, Nebraska and delivered to me by * Jim Hill. I found his report interesting, although I do not concur with his classification and interpretation of some of the liver lesions. Per your request, I am also enclosing a copy of Dr. Kimbrough's findings and report on Aroclor 1260 in the rat which you sent to me earlier in the year. Sincerely, DEG/ji enclosures Donovan E. Gordon, D. V. M. , Ph. D. Section Head, Pathology 3 deposition } EXHIBIT I LgV<U5K/ta-<3k 00QCG3 HARTOLDMON0023308 U IX Z \ I B l T i !1I HARTOLDMON0023309 . ^ ,-v -`A tit INTER-OFFICE CORRESPONDENCE MtM 1IO-TIST tJmmkrn* An rnt DacniUr 24, 1175 u+mg*. PCS Paper* 003 George Lrvinskas ha* lome questions regarding the nathotoyy section of the PCB papers. He wants to visit Northbrook alter Jan. 1 at a time convenient to 7011. Please make the necessary arrangements. GLK/lam G. 1^. Kennedy ^ '.A) 'OOP . n-v'iA > s.- ; G00CG1 HARTOLDMON0023310 UJXX ( V HARTOLDMON0023311 - INTER-OFFlCeCORRESPONDENCCj o. . ro MLK. ck.-SU#P1 . r* ' &*! ..kC J3 i'X- ---------- Dr. U*lnnin'nwttfbi>f **? `****&)?. -.,. * \'* , 'i '"**' '*"'' "h-,.V*r. a. v tha pathology melto,onn aiitnlhwa Pr-CwD papara. ^ -; -> y, \ . V" : ^ '^vr- vv-r-/:iv;--v;V ,--.; ^ t . -I!,, ; ..% . i- * Si. "A S -*.. ' ii Va ' ' . January U W* Cv , . "f' i ' " 'W Dr. Lavlnakaa ha. ra.ebadul.d hit trip far Thurwlay. January 1J. - a HARTOLDMON0023312 UJXX I B I T # ( I) HARTOLDMON0023313 ) Toxicity and Environmental Effects of Commercial PCBs Introduction . Polychlorinated biphenyls (PCBs) are not a single chemical entity. They are produced by chlorinating biphenyl to achelve a final product with specified properties. These products, sold under the trade name AROCLOR by Monsanto, are mixtures of chlorine-containing biphenyls with different numbers and positions of the chlorine substituents. Over the years, a series of Investigations have been conducted to assess potential health and environmental hazards of PCBs. The 3 predominant commercial mixtures (AROCLORS 1242, 1254 and 1260) have been studied most intensively. Toxicity' tests performed on these 3 commercial mixtures were typical in scope of those designed for the evaluation and establishment of the safety of direct and indirect food additives. It may be noted that AROCLOR 1260 no longer is produced in this country since it does not meet the physical specifications for its restricted uses. Conclusions Based on Monsanto Studies 1) As a class, the AROCLORS are relatively harmless materials for routine industrial handling under ambient conditions. 2) Threshold Limit Values of 1 mg/rn^ and 0.5 mg/m^ have been established for materials containing average chlorine values of 42# and 54#, respectively, of the available sites. 3) Tfre no-effect level for these materials in chronic 2 DEPOSITION I I EXHIBIT I 1 S'3n-8i toicl SCH 019639 C\f\C\C.l jo HARTOLDMON0023314 rat and dog feeding studies is about 10 ppm in the diet. No hepatocellular carcinomas were present. 4) The no-effect level on rat reproduction is between 1 and 10 ppm in the diet since higher levels result in low mating Indices. 5) No teratogenic or mutagenic effects were observed in studies with rats and mice. 6) In chicken reproduction and teratology studies, AROCLOR 1242, which produced the most severe effects, had a no-effect level of 2-4 ppm in the diet. 7) Acute toxicity to fish Varies from less than 1 ppm to greater than 100 ppm depending on the specific AROCLOR and species of fish tested. 8) PCBs containing 3 or less chlorine substituents per molecule are reasonably biodegradable. Summary of Monsanto's Long-term Toxicity Studies on Commercial PCBs. These tests consisted of 2-year (lifetime) feeding to rats, 2-year feeding to dogs, 3-generation rat reproduction studies with 2 litters cast per generation, rat teratology studies and dominant lethal mutagenic studies in mice. Toxicity and reproduction studies in chickens were performed to evaluate possible untoward effects In birds such as decreases in eggshell thickness. In addition, biodegradation and tissue accumulation studies have been conducted. The highest dietary level (100 ppm) in the lifetime rat feeding studies produced a weight depression at 24 months in females fed AROCLOR 1254 and liver weight increases in all SCM 019640 ' 000CG7 HARTOLDMON0023315 / groups except males fed AROCLOR 1242. As with other halogenated hydrocarbons, the important histopathologic changes were present in the livers of animals sacrificed at the end of the study. These consisted of hepatocellular alterations svlch as focal hypertrophy, cytoplasmic lipid changes and in some animals from the 100 ppm groups, hepa tomas or cholangiohepatomas. No evidence of hepatocellular carcinogenicity of the AR0CL0RS was found. In the dog 2-year studies, some slight depresses in body weight gain were noted. At the highest feeding level (100 ppm), AROCLOR 1260 produced an increase in serum alkaline phosphatase activity and liver weights without concomitant - histopathologic changes. However, there was evidence of gastrointestinal inflammatory lesions and ulcerations which appear to be similar to those found by Allen in rhesus monkeys fed AROCLOR 1248. (AROCLOR 1248 was an experimental product which never was commercialized.) In the rat reproduction studies, none of the AR0CL0RS produced adverse effects in the 2 litters of the first gen eration.* In the second and third generations, there was a reduction in the mating index at the 2 highest feeding levels (10 ppm and 100 ppm). The ability of females to conceive, carry the delivery process to parturition, and to successfully nourish the young was not affected by the 3 AROCLORS. No changes were produced in the reproductive tracts of either male or female rats by any of the 3 AROCLORS. There was no evidence of teratogenic or mutagenic changes with any of the PCBs at maximally tolerated dose levels in SCM 019641 00QCG3 HARTOLDMON0023316 u) rats and mice. In the chicken toxlclty/reproduction study, AROCLOP. * 1260 was without effect at all test levels. AROCLORS 1242 and 1254 at 100 ppm In the diet decreased egg hatchablllty. In fact, poor hatchablllty of eggs was found from hens fed 8 ppm of AROCLOR 1242. In addition, AROCLOR 1242 at 10 and 100 ppm and AROCLOR 1254 at 100 ppm were associated with reduced eggshell thickness. Chick viability was affected ' by both substances at &-dietary level of 10 ppm. Biodegradation studies were conducted using semi-contlnuous activated sludge systems and tissues of rats fed PCBs were analyzed to measure accumulation and retention of these ma terials. These factors are influenced by the number and ` position of the chlorine substituents. Higher chlorine-con taining members are more resistant to biodegradation and they accumulate more readily and are less readily metabolized and/or excreted from lipoid tissue in animals. PCBs containing 3 or less chlorine substituents per molecule are reasonably biode gradable. Comments The conclusion that "No evidence of hepatocellular car cinogenicity of the AROCLORS was found" in the 2-year rat feeding study was reached only after extensive re-evaluatlon of the original liver slides as well as additional liver sections from all of the animals after the Kimbrough results on AROCLOR 1260 became known to us. The slides were read SCH 0196<2 ooocco HARTOLDMON0023317 (J f independently by Dr. D. Gordon of Industrial BIO-TEST Labora tories, Professor W. Richter of the University of Chicago, and by Professor P. Pour of the Eppley Institute for Research in Cancer. In addition. Dr. Pour evaluated the Kimbrough slides and does not agree with the reported findings. Barsotti and Allen have reported that 2.5 ppm of AROCLOR 1248 in the diet of rhesus monkeys adversely affected repro duction. This approximates a dosage of 100 vig/kg/day. It is higher than the safe human intake (1 pgAg/day) estimated by PDA from human data when it promulgated its tolerances for PCBs in food. Oft SCH 019643 onor.70 HARTOLDMON0023318 o) . . Human data (F.R. July 6, 1973. p. 18097, Section (2)) Lowest level of PCBs producing effect In man 500 mg total. 500 tag consumed by 50 kg Individual over 50 days. 500 cog 4- 50 kg 4. 50 days 200 ufiAg/day (effect level) _ Using 10-fold safety factor leads to estimate that 20 pg/kg/ day may be-safe over a 50-day interval. Since PCBs have long half-life; calculating same Intake over a 22-month span arrives at an estimated safe intake for a pro tracted period of 1 ugAs/day. Primate data (Fed. Proc. 2^:338 (1975). Abstract 675. Barsotti & Allen) At 2.5 PPm in diet, primates Ingested 182 mg over 52 weeks. 182 mg n 365 days 0.5 mg/day which produced effects. Assuming 5 kg primate, 500 pg 4- 5 kg = 100 ug/kg/day Conclusions: Primate study done at dosages (100 ug/kg/day) greater than the safe dosage (1 ugAg/day) eslmated from human data. Primate study does show effects at lower dosages over longer time span (100 ugAg/365 days) than had been observed in man ' (200 ugAg/50 days). The tooal amount of PCBs producing effects in primates (18^ mg) was lower than that producing effects in man (500 mg). (N.B. Abrahamson & Allen, Env. Health Perspectives. June, 1973, bl-66. Report that infant monkeys are able to tolerate doses of PCBs that produce extreme morbidity in adult monkeys.) Dog and Rodent Data (F.R. July 6, 1973. p. 18097, Section (1)) nNo-effect" level for man using a 100-fold safety factor and accepting 10 ppm as a "no-effect" level in animals would be 2.5 ugAs/day based on dog data and 3. pg/kg/day based on rat .. - ' '' -- ---------------- If rat data "no-effect" level drops to 1 ppm, then corres ponding estimate of human "no-effect" level drops to 0.3 v&/ kg/day. Question: In a "collision" between rat data using a 100-fold safety factor and human data using a 10-fold safety factor, which data base would you like to be riding cn? Comment: Since human data is available, it could be argued 'that the traditional 100-fold safety factor is not necessary. Application of a 30-fold safety factor to rat data, supported by dog and human data, makes present estimate of safe human intake appear reasonable. SCM 0196<<r 000C.71 HARTOLDMON0023319 1 'i r ) PCB Primate and Human Residue Data Cone. In diet, ppm Intake, ygAg/day Estimated safe dose, USAg/day Liver eone.,uS/S Eat cone., ug/g Primates 2.5 100 56.3(0.01)* 50,0(27.7)* Value in ( ) from Infant primate. Allen et al., TAP 0: 440-451 (1974) **Yobs, Env. Health Persp.,79-81, April 1972 Human -/ 0 |3l4 samples ** <1 |125 samples 1-2 2.65 samples >2 [33 samples) SCM 0196^5 00007 HARTOLDMON0023320 V E X H I B I T / HARTOLDMON0023321 oiicnyMr ClCtaCCS encHiti** coic*tCtteCi*c<kCc*ct # *; V- Georgpe J. Levinnkaa, Ph. D* jV vlv-' s%r`- Mona 800 N. Lindbergh lUvil. '*M '. '# ^ St. Lo-u*i-e, M*is-s..o..u..r.i WI66 ^ Dear Dr. LovinskAs:s . ,* V'V5' .: . . .! * - . . r-. '' ... . V>"- . * . '. ` . * . .'% t .'V _ ^ v""..*" ' '- . ' I aan <*nelosing tlw rcvi*.*l"hinir tableaforlbe'Aroelor paper that wo discusmd during your %i*it on January 15, 197<>. .You will .. ; _ notice there arc two table* with the smite flats* _ One^table-T* a Hating of tumors by organ system, as you aucgnatcfl, to replace' the original table. I think I also prefer the f..rnu;r although it will requireranoro space when printed. . . \ J &. 'a * # * . *. : .* ..x / r # Should yuu haver any further qwli*nr# please contact me, A - -.% . & 'x.\ . ` ' > v ' ' Sincerely youra. ^ ^ * \' jr./v *:/ $ ' t)<nnvnn K. Cordon* PeV.XU* Ph. D, - Section lluad !*athulogy DfcIC/ji : cc: Me L. . ' '+ *- . * ' * V. "*i > ,' :SSTk.-*...-r,* fy , :.% :? : V rv.; > _? i v\j"V" T r --- i DEPOSITION ? EXHIBIT |LVl05MS-3c| Vi??. At. -Ar oonr^'T HARTOLDMON0023322 E X H I B I T (' HARTOLDMON0023323 m Monsanto Dipt. *>f Medicine & Environmental Health** / * ' Levinskas, A2SA ' \ November 17, 1978 AROCLOR 1260 MMMi TO i FILE . - G. Roush, Jr. - A2SA J.C. Weber - B2SK G.F. Barton - A3NC W.J. McCarville - A3NC C.F. Callis - B2SA J.T. Garrett - A2SA DEPOSITION i EXHIBIT dJSUftS* 3! On Wednesday, November 15, 1978, I had a call from Joan 1 Ranson, an industrial hygienist with OSHA in Philadelphi uxl. About two weeks ago, a Philadelphia trucking firm picked up two used transformers Westinghouse was having shipped. The truck carried other cargo of a consumer nature, namely shoes, toys and candy. The transformers were not accepted at the receiving end because their contents had spilled. Consequently, the driver took them to the truck terminal. The transformers contained AROCLOR 1260. An estimated 168 gallons of transformer fluid was spilled. CPSC has been tracking down the consumer products. Most are believed to have been recovered. EPA is on the scene. Details as to exactly what they are doing were not discussed at length. They apparently are offering advice on cleanup of the spill. Two local TV stations have camera crews at hand, and the emotional level is rising. The two leaking transformers have been removed to an enclosed area in the truck terminal. Ms. Ransom wanted to know about monitoring air levels in the enclosed area for PCBs. She also inquired about clinical or other tests which could be done on exposed workers to gauge their exposure and the nature of protective equipment which clean-up crew members should wear. SCM 0 1 9 6 3 7 Without reiterating all aspects of this somewhat lengthy discussion, I made several statements to Ms. Ranson. She was told that there were no tests we know of to measure the after effects of exposure, and none should be necessary since there was no reason to expect that persons exposed under the conditions she had described would become ill from PCBs. With respect to air monitoring, the analysts should be careful to identify PCBs as such since AROCLOR 1260 was mixed with another chlorinated hydrocarbon. The latter was more volatile and, if detected, and expressed as PCBs, could create unnecessary concern in addition to being misleading. I saw no reason for concern about the health of employees' families from possible contact with PCBcontaminated workclothes. With respect to the emotional tensions, I suggested that they tell people there was no health hazard involved, but that efforts were being made to limit the potential environmental effects of PCBs. The use of disposable or protective clothing was justified to keep workers from taking soiled garments home and having PCBs flushed down the drain. THere was no need for respiratory protection for cleanup crews. Finally, if the truck had a wooden floor-bed, it IN-10M IREV. 3/78) 000C74 HARTOLDMON0023324 FILE - AROCLOR 1260 November 17, 1978 Page - 2 - ) might be advisable to replace it. The loading dock apparently is concrete. All wastes, including spillage initially swept up with sawdust, should be disposed of in an acceptable manner according to prevailing regulations. Ms. Ranson appeared to be aware of those requirements. I don't know how many places have been contacted. Ms. Ranson stated that they had called Dow in the mistaken belief that they manu factured PCBs. They received a suggestion that adipose tissue assays might provide a useful measure of exposure. My comments on that suggestion ended with the observation that obtaining fat samples by biopsy would be the most traumatic part of the entire episode for the individuals involved. Ms. Ranson expressed appreciation for my remarks and indicated that her supervisor, Mark Durham, Sr. Industrial Hygiene Supervisor probably would call me tomorrow - Thursday, November 16, 1978. . Let's see what happens. . /bks Ge _ skas SCM 019638 HARTOLDMON0023325 V E X H I B I T # HARTOLDMON0023326 Monsanto G. J. Levinskas - A2SC September 25, 1975 PCBs TO G. Roush, Jr. A2SA H. S. Bergen - B2SL D. R. Bishop - BIND W. B. Papageorge - B2SK R. G. Potter - B3SA W.W. Withers - B2SA The attached represents a final (?) version of the toxicity statement on PCBs. This takes note of all the comments which have been received since the version mailed to you on August 29, 1975. This was discussed on the phone with Wayne Withers, and he agreed that it was acceptable. Since Wayne was the only one who had comments regarding the August 29 version, this should be satisfactory to all concerned. /bkp att. Geo w . Levinskas SCM 019716 000C7G HARTOLDMON0023327 PCBs Recently, we were informed that liver carcinomas were observed in female Sherman strain rats fed AROCLOR 1260 for 20 months. This prompted us to re-examine livers from male and female rats of the Charles River strain which had been fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 1260 for 2 years in earlier studies conducted for Monsanto Company. There are 4 elements which are closely interwoven in this matter: (1) differences in test procedures, (2) differences in results obtained by various investigators, (3) definition of what is a cancer, and (4) evaluation of potential risks, if any, to man. These will be summarized briefly. - (1) Several animal studies have been conducted with various brands c-f' PCBs. In some studies, the test material has Been identified by trade name (AROCLOR, KANECLOR). In others, there was Just a general reference to PCB. Consequently, the quality of test material with respect to the amounts and nature of contaminating impurities or by-products cannot be determined in case. In addition, several strains of test animals were used, the duration of the experimental periods varied, and there was a wide range in the depth of detail with which the observa tions wei;e reported. Consequently, it is difficult to make comparisons between these studies. . (.2) .In general, studies have shown mice to be more susceptible than rats and females to be more sensitive than males to the liver effects of PCBs. Beyond these generalizations, the results have not been consistent. Some investigators have -- . SCM 0197 IT onor.'?? HARTOLDMON0023328 reported liver carcinomas. Others have observed only- benign tumors. Several have noted changes in liver tissue without detecting tumor formation. For the reasons Just cited, it is difficult to determine the bases for these different results. The most direct comparison can be made between the 2 studies on AROCLOR 1260 since the same high dosage level of the same lot of AROCLOR 1260 was employed in both experiments. In the studies reported to us, liver carcinomas occurred in approximately 8# of female rats of the Sherman strain (the only sex used). In Monsanto's study, none of the liver lesions had progressed beyond the stage of benign tumors (hepatomas) despite a slightly longer duration of feeding of AROCLOR 1260 to rats of Sprague Dawley, Charles River strain. The Monsanto study employed rats of both sex and it did confirm the previously noted greater sensitivity of female rats'to liver effects of PCBs. (3) A review of the liver alterations seen in all 3 AROCLORS in Monsanto's studies shows that the lesions were benign in character in the traditional pathologic sense. An evaluation of all Information available to us, including the contradiction . between the recent data showing AROCLOH 1260 to be a carcinogen and our earlier negative results on the same product, leads to a conclusion that AROCLOR 1260 is not carcinogenic to all commonly used strains of laboratory test animals. `~ (4) In 1972, the manufacture of AROCLOR 1260 was dis continued in the United States and the sale of AROCLORs was restricted to a single use, i.e., as dielectric fluids. As such they are used in closed systems which will at least mini mize additional environmental contamination. Considering the ' SCM 019716 " 00007a HARTOLDMON0023329 high degree of fire risk associated with this use, and recognizing that AROCLOR 1260 may have a- weak carcinogenic potency which has not been fully proven and that AROCLORS 1242 and 1254 have not been shown to be carcinogens-Jit is ________ :------------------------------------------------------------------fS^iZST concluded that the continued use of AROCLORS^as dielectric fluids will not present an unreasonable human health hazard SCM 019719 QOOC/79 HARTOLDMON0023330 4 bee V. B. Paoageorge - B2SX & January Ik, 1973 Dr. Donovan I. Gordon Industrial HO-TSST Laboratories* Ino. 1810 Frontsgo Boed Borthbrook* Illinois 6oo6* . Bat Polychlorinated Biphenyl (PCI) Dear Deni V ; . *- '4 .* . Inclosed ara cop las of throe rsprints dealing *ith livar tuaorlgenlels of FOB'S. These aro front * . (1) Oann. (1972).1* 805. (2) Oann. (1973).6*l 105-106. ... (3) J. Bat'l Cane. Inat.. (1973).5li1637-16*6. ' Tbs studios mere performed withJapansss materials (Kanechlora), but tho asmnants aro of direct lntarsst to our naatlng oq^AROCLCR 1260y You and Dr. Blchtar should ba familiar altn-tba contents of thoaa papara as you ravisa tha AROCLOR slldas. Our meeting is schadulad for January 31* 1975 In Boon 2011 Building 37, at BCZ la Batbasda. Va *111 ba naatlng with Drs. Kimbrough and Squlrss. Bo apaelfle time has baan set, but vs eaa plan on starting in tha morning. I *111 con firm a apacific time and *111 chock *lth you or Xap on s place to stay tbs night bafars. Sincerely, ml* cc Dr. M. L. laplinger Oaorga J. Levlnakaa, Ph. D. Manager. Knvlronmantal Assess ment and Toxicology 2 DEPOSITION 1 EXHIBIT /3 SCH 022820 000(Vi0 HARTOLDMON0023331 VE X H I B I T # 3Ji HART OLDMON0023332 Monsanto *M uSilef 1 '. - TO t U Bishop - BIND i 'i^o : /x4W. Nov. 17, 1975 W. R. Corey - JS? ^Mr. F. J. Fitzgerald - B2SA 2 Dr. J. F. Mietire - T2F ^3 7 Air. D. Wood - B2SD2.o*i>^ -,713. -'Mr. T. H. Bottini - B2CA J Bi. P.-OT'DeGcUiuu B3TIA ^Dr. W. C. Hammann - B3SA*"' -Mr. E. H. Harbison - DlK *,9t Dr G. J/'Mr. W. B. Papagaorga-- B2-SK ^Or. C. Paton - B2SC .Mr, R. G. Potter - B2SB Dvr Q. Roush AT5A ^Mr. J. C. Weber - BIND J'1 7 ^Mr. W. W. Withers - B2SA-2''3'' gg.'T. HE.-Wright A2SC Attached is the second draft of a news release summarizing Dr. Pour's re-evaluation of Dr. Kimbrough's study. It incorporates comments- from Dr. Levinskas, Messrs. Papageorge and Wood. Please let me have your comments and/or approvals as soon as possible tomorrow morning. We need to clear the final version with Dr. Pour and have copies printed for distribution in Chicago on Wednesday, following Dr. Kimbrough's presentation.' I'll have to print the news release late tomorrow morning. Yovv cooperation is appreciated. /$ /inks attachment! * D. R. Bishop ^ fa 1(&k & k**4**eS ; deposition exhibit <-gy<N Uu\s-`S-zn-xi oJt SCM O58058 IN 10 *CV. ` GO0C30 HARTOLDMON0023333 xxm I I V / * HARTOLDMON0023334 imom .n*mi . V<v*i.mm D. R. Bishop * BiND OA'I Nov. 17, 1975 luH I TO Mr. T. H. Bottini - B2CA Dr. P. 0. DeGarmo - B3NA Dr. W. C. Hammann - B3SA Mr. E. H. Harbison - DlK Dr. G. J. Levinskas - A2SC Mr. W. B. Papageorge - B2SK "Mr. W. R. Corey - B2SA Mr. F. J. Fitzgerald - B2SA Dr. J. P. Mieure - T2F Mr. D. Wood - B2SD Dr. C. Paton - B2SC Mr. R. G. Potter - B2SB Dr. G. Roush - A2SA Mr. J. C. Weber - BIND Mr. W. W. Withers - B2SA Dr. P. L. Wright - A2SC Attached is the second draft of a news release summarizing Dr. Pour's re-evaluation of Dr. Kimbrough's study. It incorporates comments- from Dr. Levinskas, Messrs, papageorge and Wood. Please let me have your comments and/or approvals as soon as possible tomorrow morning. . We need to clear the final version with Dr. Pour and have copies printed for distribution in Chicago on Wednesday, following Dr. Kimbrough's presentation. I'll have to print the news release late tomorrow morning. Your cooperation is appreciated. ' /Q- /mks attachment D. R. Bishop * deposition f EXHIBIT 10 <*f. 1 - SCM 058337 cnoooi HARTOLDMON0023335 :- IMMEDIATELY 1975 _ "" . . MONSANTO INDUSTRIAL CHEMICALS CO. D. R. Bishop (314) 694-2891 PUBLIC RELATIONS OEPARTMENT 800 N. Lindbergh Boulevard SL Louie, Mlsaourt <3166 CHICAGO, Nov. 19 -- Monsanto Company today announced that a newly completed scientific report, commissioned by the St. Louis-based company, does not confirm the presence of malignant liver tumors in experimental rats fed a commercial polychlorinated biphenyl (FCB) mixture...a conclusion that had been previously reached and widely reported by Dr. Renate D. Kimbrough of the Center for Disease Control of the U.S. Public Health Service in Atlanta. The Monsanto-sponsored report, a re-evaluation of Dr. Kimbrough's work, is titled, "Histopathological Re-evaluation of Tissues from Sherman Rats Fed Aroclor 1260," and is authored by Dr. Peter Pour, a pathologist with the renowned Eppley Institute for Research in Cancer of the University of Nebraska Medical Center . at Omaha. Aroclor is a Monsanto trademark for a class of chlorinated hydrocarbon Industrial chemicals it manufactures. In her study. Dr. Kimbrough reported that when Sherman strain female rats were fed 100 parts per million of PCB (Aroclor 1260) for about 21 months, 170 of the 184 experimental animals developed neoplastic lesions (precursors of malignant tumors). She further identified 26 of these lesions as hepatocellular carcinomas (malignant liver tumors) . SCM 05 8 33 8 onocn^ HARTOLDMON0023336 --2 During his re-evaluation, Dr. Pour saw the same alteration but identified them as being hyperplastic or non-tumorous lesions, further pointing out that it was his strong impression that many of the detectable alterations were a degenerative and reparative rather than a neoplastic process. Dr. Pour's re-evaluation is consistent with conclusions drawn from Monsanto's own PCB feeding studies which have never produced a carcinogenic response in experimental animals. . In his discussion, Dr. Pour reported that he observed 20 cases of abnormal lesions which presented such structural criteria as to be considered possible precursors to tumors, although a most significant criteria for malignancy -- namely invasiveness -- was missing, and the sign of ongoing toxic, degenerative and regenerative processes in the rest of the tissue was evident. "At present, the statement that these lesions may have metastasized (infiltrated adjacent tissue) if the treatment had been continued is as much as unreliable as the possibility that they might have been regressed if the animal would have been kept longer or for life," he stated. "In summary," the Eppley Institute pathologist reported, "It is difficult at present to conclude whether or not some of the examined lesions represent a malignant lesion, because of the lack of invasion and metastases. In the case of such organs, as the liver, with its marked tendency toward regeneration, it is, in my -more- SCM 058339 0^0C03 HARTOLDMON0023337 --3 opinion, difficult and sometimes impossible to distinguish between regeneration, hyperplasia and neoplasia, particularly when the tissue is continuously exposed tp a toxic substance." Quoting further from Dr. Pour's report, he wrote, "in this context. Dr. Kimbrough's study seems of particular interest, in that polychlorinated compounds may be retained in the body, even in higher concentrations for several months after feeding has been stopped. This finding may explain the continuing degenerative and regenerative processes in the livers of most experimental rats, even two months after Aroclor was removed from the diet. "One should also bear in mind that some animal strains may react more specifically to a compound because of common endogenous diseases, as in the case of the Sherman rats used in this experiment which tend toward spontaneous liver lesions. "In my opinion," he concluded, "many of the lesions induced were of a degenerative nature, and without further studies, the induced liver lesions could not be definitely designated as neoplasia (tumorous) . -0O0- NOTE TO EDITORS: Copies of Dr. Pour's report are available on request, in writing, from Public Relations Department, Monsanto Industrial Chemicals Co., 800 N. Lindbergh Blvd., St. Louis, Mo. 631 SCM 0583^0 .GOOCG* HARTOLDMON0023338 V \ E X H I B I T i HARTOLDMON0023339 7oxicoi.oar CMCMIITXT MCOICAL SCIKNCCa MiCMOaiouear iNoutr.uL iaxcty * mcaltm BIO -TEST Jbd^yioJ/yiuzb, 3/x. 1810 FRONTAGE ROAO ' NORTH8ROOK,ILLINOIS 80082 July 31, 1981 . A*CA COOt 311 TCUXHONt 171-3010 TCLXX 71---17 Or. George Levinskas Monsanto Company 800 North Lindbergh Boulevard St. Louie, Missouri 63166 Dear Or. Levinskas: As per your request of July 30, 1981, Or. O. E. Gordon is attempting to locate the raw data for the pathology reports on histologic examination of additional liver sections from rats fed Aroclors 1242, 1254 and 1260 (IBT study no. 641-06672). These data will be microfilmed and a diazo copy sent to Mr. A. Uelner, Manager, Quality Assurance. Xerox copies of these data will be mailed to your attention. If I can be of further help, please contact me. Sincerely J Marilyn A. Biederer Validation Assistance Specialist MABjAR cc - Mr. A. Uelner Monsanto Company SCM 023698 HARTOLDMON0023340 LUXX I I B I T # ( HARTOLDMON0023341 Monsanto De, of Medicine & Environmental Health J. Levinskas, G2WF (4-83C9 July 9, 1981 AROCLOR Products 1242, Two Year Rat Feeding St a!nd 1260: J.R.G. Ortiz, E2ND A.F. Uelner, G2WC J.H. Craddock A complete audit of these three past IBT studies will require considerable time. So much time, in fact, that the results of the audit would not be available until long after completion of the monograph which is being prepared. As a result, it has been decided to limit the audit to a determination of how long animals were on test and as to whether or not liver reactions were taken for microscopy. These are the crucial elements for assessing potential carcinogenicity of these materials. GJL/mcl INOOM (H6V. 3/731 SCH 058929 HARTOLDMON0023342 UJXX I B I T / l HARTOLDMON0023343 Monsanto C- Njme ol Nominee LocJl'On Paul L. Wright Creve Cceuj Comoany Un.C at SUM Oaot. Oivition 8utmu Group 0O4ftmnt Corporate Staff rVolition i(it Medical Salary Gf Annual Salary Medical Oata of Last incraait Toxicology Manage: 19 $28,980.00 2/75 Par'ormanca Growth 0*ta lilt Employ# Avtw Excellent Excellent 2/75 Type of Award (Select the award category from tne opposite tide of (hi* meet wh<n most epprooeateiy descriOts the award end anfar tne category ano __ code oeiow.) Staff _______ Award Category Category Code Numoe Solution to product toxicity problem which permitted Monsanto 28 orib. *___i_n___*. Jjc7_hivm"an*t1 nd. _ !!' it.1s' itfniftcjn'c_____to_ Monunto otiowto contmmuisa product mmaannutlff actIuI rin^ and sale. Dr. Paul L. Wright has performed his regular duties exceedingly well and has, in addition, completed a specific "ad hoc" assignment with dispatch and distinction. Dow informed us that they had encountered lung tumors in rats fed maleic anhydride and that they felt they had to report these findings to the Pood and Drug Administration (FDA) ait once under their "product stewardship" program. Dr. Wright was asked to contact Dow's toxicologists for details of their findings. This he did, and by his personal and professional; efforts he convinced Dow personnel that while they had an obligation to report their findings to FDA, it would be foolhardy to act precipitously. Instead, Dr. Wright advised Dow to defer contacting FDA until they had thoroughly evaluated their findings and a subsequent course of action had been developed to allow resolution of questions which undoubtedly would be raised by their report of tumors. Dow agreed to this. Dr. Wright then contacted the Process Chemicals business group and informed them of Dow's findings. He proposed a dual course of action. The first : step was a commitment to support a repeat of the feeding study to determine whether Dow's findings were reproducible. The results of such a study would not necessarily resolve FDA's concern over the potential carcinogencity of maleic anhydride, but the intent to conduct the study promptly would serve to forestall precipitous action against the product by FDA. The subsequent step was to provide assurance to FDA that a thorough investigation of the toxicity of maleic anhydride would be undertaken, either ay Monsanto alone or in cooperation with others. This proposal was acceptable to the business group, and Dr. Wright so informed Dow. Subsequently, Dr. Wright accompanied Dow personnel to FDA when they presented their findings on maleic anhydride. FDA's toxicologists reviewed the data and the proposed plans of actions. They concluded that while the questions of carcinogenicity would need to be resolved, there was 10 apparent need for immediate prohibition of maleic anhydride or polymers aontaining maleic anhydride in food contact applications. Thus, we aelieve that Dr. Wright played a singular, outstanding role in preventing TDA from publicly proclaiming that maleic anhydride was a suspect Racommandad 6yt Oato Award Amount Racammandadi Acorawd 8yi 0.. Award amount aporoved lor payment: ^---------------------------------------------------------------------------------------------------------------------------------------j- / 2 DEPOSITION O-IJJJ [y | EXHIBIT 1 ^7-31 ul>i<L $2,500.00 k --------- SCH 058799 000:03 HARTOLDMON0023344 er." A ward 2::a (\. :ei carcinogen. Because of the many uses of maleic anhydride in feed contact polymers, this was a significant accomplishment. Subsequently, when maleic anhydride was included in the priority list for study by CUT, Dr. Wright undertook writing of a literature review on the toxicity of this material. Notwith standing the fact that he had assistance in locating pertinent articles in the literature. Dr. Wright had to read, digest, and write a substantive review in a relatively short time. This meant considerable extra work on nights and weekends. He completed his assignment on time, and it was a credit to him personally and to Monsanto. At present. Dr. Wright has been given the added assignment of preparing protocols for further toxicity testing of maleic anhydride by CUT. We are confident that he will complete this assignment with equal professional competence. Therefore, it is a pleasure to nominate Dr. Paul L. Wright for a merit award on the basis of his demonstrated performance. G. J. Levinskas I SCM 058800 00Q--* HARTOLDMON0023345 xxm 1. \ I B I T HARTOLDMON0023346 Monsanto f a Nam* Ol Nomina* Company unit or Staff Oept. OivtiMn Med. & F.nv. Hlth. Poftition Title . Solary Grade Toxicolocv Manager Potior mane* 19 Butineai Group Annual Salary S31.800 Growth Oapartmant Location Creve Cceu: Oate of Laft Inc/eate 2/1/76 Date Laat Employe Review Typ* of Award . c(Soadi*acbtdtlnow.a)ward catatory from in* oepoittt aid* of ml* tnaat wnlcfl moit appropriitt,ry attend** tn award and antaa tna categpry and Award Cataoory Category Cod* Nun OTaiceriOcahtnna iaccnalavlamant Accomplishment of significant and It* iinillanea to Monunto Balowi results 106 At a recent meeting in Creve Coeur, Glenn Schweitzer, head of the Office of Toxic Substances of EPA, offered some interesting comments. He observed that Monsanto's toxicologists were held in high regard at EPA. However, since it lacked an in-house toxicology facility, Monsanto as a company was not as highly regarded overall as duPont, Carbide, or Dow. Dr. Wright's professional and personal characteristics have contributed sig nificantly to Monsanto's image at EPA. He has shown unusual perseverance and dedication, frequently involving his own time, to review and interpret large volumes of data which he subsequently organized for presentation to EPA officials. Particularly noteworthy were his efforts on polychlorinated biphenyls (Aroclors) and chlorinated isocyanurates (ACL products). In the former instance, his excellent analysis and synthesis of widely scattered observations played a prominent role in forestalling EPA's promulgation of unrealistic regulations to limit discharges of polychlorinated biphenyls. EPA's proposed regulations would have precluded the use of these materials by Monsanto's customers. With respect to chlorinated isocyanurates, Dr. Wright has had several contacts with individual scientists at EPA to answer specific questions that they had raised or to inform them of the status of additional studies which, had been undertaken*. Overall, by virtue of the qualities of leadership which Dr. Wright has dis played, he has made an important contribution to Monsanto's image at EPA. That favorable image will become increasingly important to Monsanto as the areas of interaction with EPA multiply. , ^POSITION r EXHIBIT , 7- uu/ [ Racommandad Syt ' C-2LJ: |n*v. 10/74) Oat* ARwecaormd mAmenoouendti Approved Byi ------- -- rtnn-* Oat* Awapappradryomavemednotuf:onrt 7? Aft SCM 058795 HARTOLDMON0023347 E X H I B I T HARTOLDMON0023348 'woa i tG (Pv d'82) AUTHORIZATION TO RELEASE INFORMATION (PRIVATE PERSON OR ORGANIZATION) TO PROBATION OFFICER TO WHOM IT MAY CONCERN: I.---------------- Pa-.---L* ----------------------------------------- , the undersigned, hereby authorize the United States Probation Office for the EasternDistrict nf Missouri or its authorized representative(s) or employee(s), bearing this release or copy thereof, to obtain anv information in vour files pertaining to my: [if Employment Q Education Records (including but not limited to academic achievement, attendance, athletic, personal history, and disciplinary records) Medical Records Psychological and Psychiatric Records ' I hereby direct you to release such information upon request of the bearer. This release is executed with full knowledge and understanding that the information is for the United States Probation Office's official use. I hereby release you, as custodian of such records, any school, college, or university, or other educa tional institution; hospital or other repository of medical records; social service agency, any employer, or retail business establishment including its officers, employees, or related personnel both individually and collectively, from any and all liability for damages ofwhatever kind which may at any time result to me, mv heirs, family, or associates because of compliance with this authorization and request for information or any other attempt to comply with it. The information hereby obtained by the aforementioned probation office is to be used only for the purpose of presentence investigation and report and, if applicable, for supervision. HARTOLDMON0023349 PPO0 140 13/64) united l t * r e j 9iir*icT C0lt FCOCIUL ^KOSaTiOh IYITIm REQUEST FOR EMPLOYMENT OATA AOORCSS OF RROEATION OFFICE 111 U.S. Court & Custom House 1114 Market Street St. Louis, MO 63101-2071 r - Personnel Department Monsanto Company 800 North Lindbergh Blvd. St. Louis, MO 63141 L 3/7/Ri Dear Sir: The person identified below is j-der investigation by this office. The mformetion requested is needed to complete this investigation. Your cooperation will be greatly appreciated. Please return this form^aiiAifl three days in the enclosed tavq * Pat "I aooreu op person seinc investigated 2001 North Geyer Rd. J St. Louis, MO 63131 States employed fr om Nov., 19/2, to Feb. 29, 1984, as manager of special projects in Dept, of Medical & Environmental Health earning $53,000 per WAS THIS PERSON EVER EMPLOY? ____ y pjjvti POSITIONS HELO IN YOUR o / year. Please information oesmeo verify. "Y*S*IF * GIVE Oats start.co OATS LEFT U ///7 2- 9i 7-------7------ ' gS SALARY OR WAGE s . RE_ASON FOR TCRMIM ATI NO EMPLOYMENT / (y j f-- - . - >4 if WAS THIS PERSON'S SALARY ATTACHED* O ru WOULD YOU C0NS40ER REEMRLOYING THIS PERSON! REMARKS (Concmmtn4 tfite pwai'i ntediiw, IWMey, wthttHty, and dtitmm* ttamm --plered V* rw intafuMi.) AUTHORIZATION TO RELEASE CONFIDENTIAL INFORMATION form attached. 000/.07 HARTOLDMON0023350 UJXX / HARTOLDMON0023351 GMErt Cl'*-"-4 DMEH C" Toxicology Section/St. Louis C0J0tvj0frr7L0CATi0N) SPECIAL (TYPE OF REPORT) REPORT LO N -t CO H ** NO - t2--i Q-CO CQ HJ Xa. CJ = < O Urn </ oB. o rt cu oi (U tfl oi > CH OX ti--3t J>uaou :ni <H >-j B, O OH 4> ' M>- O U (-> X O4> t C* CJ rH CD XQ tooc 3 o tii 3< REPORT NO.: MSL-2 002 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: TOXICITY OF AROCLOR PRODUCTS 1242,;1254 AND 1260 TO THE LIVER OF ALBINO-RATS AUTHORS: George J. Levins leas, Ph.D. ABSTRACT: This report is a tabulation of the results of microscopic evaluation of liver sections from rats fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 12 60 for two years. A-IOMIat SCM 058930 0001.03 HARTOLDMON0023352 DISTRIBUTION COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - DMEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H.Craddock, A2SA 8 - G.J. Leviriskas, G2WF 9 - J.G. Nassi'f, E2ND '' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: REPORTS LIBRARY, R2C 1 If you transfer it to anyone else, please let your librarian know, so the records can be changed. ooo:.co R-ioaa(C) (Rev. i/ao) SCM 058931 HARTOLDMON0023353 INTRODUCTION The polychlorinated biphenyls (PCBs) comprise a family of compounds of variable chlorine content. PCBs manufactured by Monsanto (tradenamed "Aroclor") bear a four digit number, the '12' indicating biphenyl and the last two digits indicating the chlorine content by weight percent. Since the chlorine atoms are randomly distributed among the 10 ring positions available for substitution, each material is a mixture of isomers. In 1969, Monsanto sponsored a series of animal studies at Industrial BIO-TEST Laboratories in Northbrook, Illinois, on Aroclor 1242, 1254, and 1260 for the assessment of the health and environmental hazards of these materials. This series consisted of 2-year chronic feeding studies to rats and dogs, a 3-generation rat reproduction study, a rat teratology study, a dominant lethal mutagenic study in mice and a toxicity/reproduction study in chickens on each of the 3 Aroclor products. Even though no such action was contemplated, such a broad battery of tests would have been adequate to support Food Additive Petitions for each of these materials. These studies were initiated by reports that PCBs had been detected in the environment. A report (Nelson, 1972b) by a Panel on Hazardous Trace Substances of an Ad Hoc Committee on Environmental Health Research covers the development of information on the environmental and biological effects of PCBs. There have been subsequent literature reviews which need not be recapitulated here [DHEW, 1978; EPA, 1976; I ARC, 1978; NAS, 1979; Roberts, et al. (1978)]. Starting in 1969, while these studies still were in progress, information regarding them was made available to various government groups.1 Subsequently, data from these studies were presented at a conference -1- GO Q:\19 SCM 058932 HARTOLDMONOQ23354 sponsored by the National Institute of Environmental Health Sciences at Rougemont, N.C. on December 20-21, 1971, but the account of these studies was omitted from the published proceedings (Keplinger, et al., 1972; Nelson, 1972a). Subsequently, it was reported that female Sherman strain rats developed hepatocellular carcinomas when fed Aroclor 12602 from Lot No. AK-3; the same lot used for the earlier Monsanto study with this material. As a result, Monsanto requested the contract laboratory's pathologists to review livers from all available rats from the 3 Aroclor studies. That review (which could have included new sections of liver from animals examined previously in addition to sections from animals not examined previously) was presented in the reports of Gordon and Richter (1975a,b,c). In November 1975, reports containing evaluations of liver sections from the 2-year rat feeding studies (Gordon and Richter, 1975a,b,c) and the results of an independent evaluation of the same liver sections (Pour, 1975) were . presented to and discussed with several federal regulatory agencies3. Later that month, a summary of data from all of the studies was presented at the National Conference on Polychlorinated Biphenyls sponsored by the Environmental Protection Agency and held in Chicago on November 19-21, 1975 (Calandra, 1976). Only summaries of data from these and other toxicity studies conducted over the years have been published (Jenkins, et al., 1972; Keplinger, et al.. 1971; Monsanto, 1980). Knowledge of these efforts may have prompted the statement in a recent article that "...Monsanto, whose reaction to the possibility that polychlorinated biphenyls (PCBs) might be an ecological disaster was a classic of how such issues should be handled, says Ford4. Monsanto began to investigate the dangers as soon as they were 2 - -a SCH HARTOLDMONOQ23355 seriously voiced. It insisted on keeping an open mind about them. As soon as evidence appeared that there was a strong chance PCBs were a hazard, it published the results of its investigations, admitting the danger. It thus established a reputation of honesty even among the environmentalists." (Clutterbuck 1981). At a later meeting in Bethesda, MD.5, pathologists selected and reviewed some liver slides from the Monsanto-sponsored and Kimbrough studies. The pathologists differed in the terminology used to classify the lesions. They did conclude that the general incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) were greater in the rats from the study subsequently published by Kimbrough, et al. (1975). No carcinomas were seen in the Monsanto-sponsored studies. It was noted that either the strain of rat or sex could have accounted for the differences. The spontaneous incidence of hyperplastic or neoplastic liver lesions in the Sherman strain rat was unknown, and the occurrence of such lesions appears to be higher in female rats and mice. The different diagnoses were discussed with Dr. Philippe Shubik of the Eppley Institute for Research in Cancer at the University of Nebraska. Dr. Shubik suggested that the liver sections from these studies could be reviewed by Dr. Parvis Pour. This was done. This publication is an effort to make readily available information in the Gordon and Richter reports (1975a,b,c) which are only summarized in the literature (Calandra, 1976). - 3 SCH 05893<. 000 O A *+* HARTOLDMONOQ23356 METHODS It appears that the Threshold Limit Value of 0.5 mg/m3 for chlorobiphenyl with 54% chlorine (ACGIH, 1969) was used to estimate suitable dose levels for the rat feeding studies. For that purpose the following assumptions were made: if the ambient air concentration of PCBs is 0.5 mg/m3, and if all of the inhaled PCBs are absorbed, and if 15 m3 of air are inhaled during the working day, a person would receive a PCB dose of 7.5 mg/day. The corresponding dosage would be approximately 0.1 mgAg/day for a 70-kilogram person. Consequently, dosages of 0.1, 1, and 10 mgAg/day were decided upon, and the dietary concentrations were set at 1, 10, and 100 ppm. As it turned out, the assumptions used to set dosages for the feeding study resulted in the low dose level rats receiving a dosage that was 100 times higher than the 0.001 mg/kg/day which FDA later calculated as allowable for protracted ingestion based on human data (FDA, 1973). For the chronic toxicity study in rats6, weanling animals of the Charles River CD strain7 were divided into a control group and 9 treatment groups, each consisting of 50 males and 50 females, with 3 treatment groups assigned to each of the 3 Aroclor products studied (1242, 1254, and 1260). Not all of these animals started at the same time. After about 2 months on test, additional groups of males and females were assigned to each test diet and the controls. These animals were added to allow a sufficient number of animals for sacrifices at 3, 6 and 12 months to provide some information prior to the completion of the 2-year study period. The uumbermg- 6equence SCM 058935 OOO:.'.3 HARTOLDMON0023357 and^-thd.^4esicj.ation,^ of .some, ^animals ^as^Jl&xtra.1**-suggests that additional animals were. placj*d^on test. Groups of 5 males and 5 females were scheduled to be sacrificed after 3, 6, and 12 months of feeding, and survivors were to be sacrificed at the end of the test period. Animals were to be given a gross autopsy and tissues were to be preserved for possible histologic examination. Tumors or lesions suggestive of tumors were to be examined. RESULTS Data from the Gordon and Richter reports (1975a,b,c) on liver slides are shown in Tables 1, 2, and 3 for Aroclor 1242, 1254, and 1260, respectively. While the text of the reports contained comments specific to the particular Aroclor, they also contained similar comments such as "There is evidence of a chemical effect on the liver.......... This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoma" and "In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study". The reports also stated that "The other treatment-related lesions reported are regarded as degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotrans formation of the test material" and "In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report.......... No hepatocellular carcinomas were observed". * 5 ' `* SCM 058936 <no:u* HARTOLDMONOQ23358 With respect to Aroclor 1254, it was noted that "One liver tumor (a hepatoma) previously reported in a T-II animal (No. 445) was re-classified as nodular hyperplasia" (Gordon and Richter, 1975a)10. DISCUSSION The initial reports of these studies concluded that the target organ was the liver for each Aroclor product. This was confirmed by the second evaluation of liver slides (Gordon and Richter, 1975a,b,c). These alterations were slow to develop, their incidence being related to both dose level and duration of treatment. Since there were increased incidences of vacuolar changes in the cytoplasm of the hepatocytes and focal hypertrophy at all dose levels for all 3 Aroclor products at the 24-month sacrifice, a "no-effect" level was not established for liver effects. With respect to the slides from Industrial BIO-TEST, Pour (1975) concluded that Aroclor 1242, 1254, and 1260 "showed a dose-dependent hepatotoxic effect, characterized by degenerative and regenerative processes. With one exception, all lesions were considered non-neoplastic. Structures similar to cholangiocarcinomas and hepatomas were found in one rat. However, the possibility of metastases of a carcinoma into the liver has been also considered." In the report, he noted one lesion which seemed to "represent a metastatic tumor (probably a mammary gland carcinoma of the same animal from which ...[the particular liver section]... was submitted), rather than a cholangiocarcinoma". This occurred in an animal fed 100 ppm of Aroclor 1254. The contract laboratory pathologists diagnosed the presence of mammary tumor metastases in the liver of this rat (Gordon and Richter, 1975b).11 SCM 058937 (no:'.:5 HARTOLDMONOQ23359 SUMMARY and CONCLUSIONS Groups of male and female rats were fed diets containing either 1, 10, or 100 ppm of one of 3 Aroclor products, 1242, 1254, or 1260, for 2 years. Focal hypertrophy of the liver occurred at 3, 3, and 12 months for rats fed Aroclor 1260, 1254, and 1242, respectively. Focal hypertrophy was not seen in livers of control animals. At the 24-month sacrifice, livers of animals from all dose levels of the 3 Aroclor products had an increased incidence of vacuolar changes and focal hypertrophy. Livers of some animals fed 100 ppm of each Aroclor also had benign liver tumors (hepatoma or cholangiohepatoma). No hepatocellular carcinomas were observed. 7- SCH 058938 ono HARTOLDMON0023360 Footnotes Dates of initial correspondence and contacts. October 3, 1969. E.P. Wheeler to A.R. Glasgow, Division of Pesticides. Food and Drug Administration. April 8, 1970. R.E. Kelly to H. Bluraenthal, Petitions Review Branch, Food and Drug Administration. ' April 22, 1970. E.P. Wheeler to E.J. Burger, Jr., Office of Science and Technology. June 1, 1970. E.P. Wheeler to H.E. Stokinger, Bureau of Occupational Safety and Health. R.D. Kimbrough, personal communication. November 13-14, 1975. Council on Environmental Quality. Environmental Protection Agency. Food and Drug Administration. House Subcommittee Manpower, Compensation, Health and Safety. National Cancer Institute. National Institute for Environmental Health Sciences. National Institute for Occupational Safety and Health. Dr. David Ford of Bath University. . At National Cancer Institute on January 21, 1975. Present were D.E. Gordon, R.D. Kimbrough, G.J. Levinskas, R.A. Squire, W.R. Richter. .' Since the events described earlier, the validity of many toxicity studies conducted by Industrial BIO-TEST Laboratories has been challenged. Therefore, the available raw data supplied by Industrial BIO-TEST was reviewed to determine whether this study could be validated. The review showed that the data bases (including the lack of a protocol) were insufficient for a complete validation of the study. It was decided to focus on presenting the primary liver effects reported by Gordon and Richter (1975a,b,c). Therefore, a complete audit was not undertaken. Available records were examined for a determination that the animals were placed on test and of their ultimate fate. Necropsy and microscopic reports were also examined for findings pertaining to livers of those animals. Significant discrepancies which were found between data in Tables 1, 2, and 3 and the data base for the Gordon and Richter reports are noted in this report. On some records, the labels Aroclor 1242 and Aroclor 1260 are interchanged as determined by a check of the animal numbers. Charles River Breeding Laboratories, North Wilmington, Massachusetts. Animals sacrificed at 6 and 12 months were placed on test about 2 months after the first group. Those sacrifice periods are sometimes labeled 8 and 14 months, respectively, which corresponds to the calendar time from the start of the test but overstates the time on test for those groups. 8- SC* ooo 17 HARTOLDMONOQ23361 9. As a result of the examination described in footnote 6, a few animals were reassigned to other dose levels on basis of assigned numbers. Three with no recorded liver lesions were deleted from the 24-month listing because of short duration on test: 14 months at 100 ppm Aroclor 1242, 15 months at 10 ppm Aroclor 1254 and 13 months at 100 ppm Aroclor 1254. Therefore, numbers in Tables vary slightly from those in reports of Gordon and Richter (1975a,b,c). 10. That change and comments in the footnotes to the Tables were noted ' during the examination described in footnote 6. In addition, the following also were noted during the examination. Aroclor 1254 - 100 ppm animal marked "Extra3" with diagnosis of hepatoma in record of examination for original Industrial BIO-TEST report. Animal not listed in Gordon and Richter report. . - 100 ppm animal no. 542 with gross autopsy notation that liver appears tumorous and white foci has no record of examination. Aroclor 1260 - 100 ppm animal no. 293 with gross autopsy notation of tumor on liver has no record of examination. . In some instances, diagnoses were crossed out on the available records. These were included in the Tables. Crossed out diagnoses for hepatoma or cholangiohepatoma are noted in the footnotes to the Tables. ' 11. Dr. Pour's report does not include the following liver sections noted by discrepancies between his tabulation and the Gordon and Richter reports. 1 from control at 12 month sacrifice, 5 from 100 ppm Aroclor 1242 at 24 months, 1 from 100 ppm Aroclor 1260 at 3 months. None of these animals were reported as having hepatomas or cholangiohepatomas in the Gordon and Richter reports. SCM 0589^0 onn''^ HARTOLDMON0023362 References 1. ACGIH (1969). Threshold Limit Values of Airborne Contaminants. American Conference of Governmental Industrial Hygienists. Cincinnati. 2. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. In: National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. EPA Report No. 560/6-75-004. March. . NTIS PB-253 248. pp.35-42. 3. Clutterback, D. (1981). What causes environmental conflict? New Scientist 90, 176-177. 4. DHEW (1978). Final Report of the Subcommittee on Health Effects of PCBs and PBBs. Environ. Health Perspect., 24, 129-198. 5. EPA (1976). National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. U.S. Environmental Protection Agency. Washington, D.C. EPA-560/6-75-004, March. NTIS PB-253 248. 6. FDA (1973). Polychlorinated biphenyls (PCB's). Contamination of animal feeds, foods and food-packaging materials. Fed. Regis., July 6, 38, 18096-18103. 7. Gordon, D.E. and Richter, W.R. (1975a). Two-year chronic and toxicity study with Aroclor 1242 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 8. Gordon, D.E. and Richter, W.R. (1975b). Two-year chronic oral toxicity study with Aroclor 1254 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 9. Gordon, D.E. and Richter, W.R. (1975c). Two-year chronic oral toxicity study with Aroclor 1260 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 10. IARC (1978). Polychlorinated biphenyls. IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. 18, 43-103. International Agency for Research on Cancer. Lyon, France. October. 11. Jenkins, D.H., Keplinger, M.L., Fancher, O.E., Wheeler, E.P. and Calandra, J.C. (1972). Reproductive effects of polychlorinated biphenyls in white leghorn chickens. Toxicol. Appl. Pharmacol. 22, 316. 12. Keplinger, M.L., Fancher, O.E., Calandra, J.C. (1971). Toxicologic studies with polychlorinated biphenyls. Toxicol. Appl. Pharmacol. 19, 402-403. - 10 - SCM 058941 mnn.' * r\ HARTOLDMONOQ23363 13. Keplinger, M.L., Fancher, O.E., Calandra, J.C., et al. (1972). Toxicological studies with polychlorinated biphenyls. Read at PCB Conference, Quail Roost Conference Center, Rougemont, North Carolina, 20-21 December 1971. Cited in Polychlorinated Biphenyls Environmental Impact. A Review by the Panel on Hazardous Trace Substances. March 1972. Environ. Res. 5, 249-362. 14. Kimbrough, R.D., Squire, R.A., Linder, R.E., Strandberg, J.D., - Montali, R.J., and Burse, V.W. (1975). Induction of liver tumors in Sherman Strain female rats by polychlorinated biphenyl Aroclor 1260. J. Natl. Cancer Inst. 55, 1453-1459. 15. Monsanto Company (1980). Polychlorinated biphenyls. PCB's. A report on Uses, Environmental and Health Effects and Disposal. 16. NAS (1979). Polychlorinated biphenyls. National Academy of Sciences. Washington, D.C. 17. Nelson, N. (1972a). Comments on research needs. Environ. Health Perspect., 1, 181-185. 18. Nelson, N. (1972b). Chairman, Panel on Hazardous Trace Substances. Polychlorinated biphenyls - environmental impact. Environ. Res. 5, 249-362. 19. Pour, P. (1975). Histopathological reevaluation of livers for rats treated with Aroclor. August 1 (unpublished observations). 20. Roberts, J.R., Rodgers, D.W., Bailey, J.A., Rorke, M.A. (1978). Polychlorinated Biphenyls: Biological criteria for an assessment of their effects on environmental quality. National Research Council of Canada. Ottawa. Publication No. NRCC 16077. - 11 - cnQ..~o SCH 0589^2 HARTOLDMON0023364 Sacrifice Interval Diet Level (ppm) Table 1 Aroclor 12A2: 'Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Termina1 a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cholangiohepa toma 10 8 10 9 202 010 000 000 00 0 00 0 000 000 1 0 0 0 0 1 0 0 10 10 8 9 1 00 1 10 030 000 000 000 000 000 0 0 0 0 0 0 0 0 10 10 10 9 1 2A 000 00 1 000 000 101 000 000 0 0 0 1 0 0 0 0 23 32 29 19 178 111 1 00 023 111 533 000 000 9 0 0 8 8 3 3b 1C * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 19, 22, 22, 23, 23 months. 10 ppm group has animals dead at 23, 23 months. ,#100 ppm group has animals dead at 22, 22, 22 months and 2 marked "Extra". b Includes 1 animal without record of examination in original IBT report. Not listed as hepatoma in worksheets for Gordon and Richter report but appears and was crossed out on typed tabulation for animal marked "Extra". q for other 2 animals do not appear in records of examinations for original IBT report. O' ' Q\ Diagnosis does not appear in records of examinations for origina'l IBT report. i: Diagnoses SCH 0 5 8 9 ^ 3 HARTOLDMON0023365 Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cholangiobepatoma 10 10 10 10 233 01 1 000 000 000 000 000 000 1 0 1 1 0 0 0 0 10 10 10 10 1 00 1 20 000 000 000 000 000 000 0 2 2 4 0 0 0 0 10 10 10 10 1 14 000 002 004 000 111 000 000 1 1 1 2 0 0 0 0 23 30 26 26 179 131 120 034 103 563 00 0 000 13 1 1 14 14 4 4b 2C a Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 22, 22 amnths, 1 marked "Extra" and 1 other for which date of death is not recorded. 10 ppm group has animals dead at 22, 22, 22 months. ,,100 ppm group has animals dead at 23, 23 months and 9 marked "Extra". b Includes 2 animals marked "Extra". Diagnoses do not appear in records of examination for original IBT reports. Qc Both animals marked "Extra" with same designation. 1 without apparent record of examination for original IBT report. q Diagnosis for other animal does not appear in records of examinations for original IBT report. t: n SCa Os,a9< HARTOLDMON0023366 Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats Sacrifice Interval Diet Level (ppm) 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Termina1 A 0 1 10 100 No. Animals Examined 10 10 iob 10 10 6 5 9 10 9 10 9 23 26 25 25 Findings Vacuolar change 202 3 11 Focal necrosis 0 0 4C 0 10 Focal lymphoid infiltration 000 0 02 Focal hypertrophy hepatocytes 000 2 00 Nodular hyperplasia 000 0 00 Ductular hyperplasia 00 1 0 00 Hepatoma 000 0 00 Cholangiohepatoma 000 0 00 *. Control group has animal dead at 19 months and 2 marked "Extra". 2 ld 2 0 0 0 0 0 6 0 0 3 0 0 0 0 1 25 000 000 000 000 1 10 000 000 3 0 0 4 1 2 0 0 15 7 9 143 6 10 1 0 0 3 13 9 107 6 5 6 7 12 0 0 le 7f.g 00 0 1 ppm group has animals dead at 20, 22, 22 months. 10 ppm group has animals dead at 19, 22, 22 months and 1 marked "Extra". 100 ppm group has animals dead at 22, 22 months. ** Includes 1 marked "Extra". c Worksheets show listings under this diagnosis with arrow pointing to Vacuolar change column. d,Date of death of this animal not recorded. e No record of examination for original IBT report. Listed on worksheets for Gordon and Richter report with diagnosis crossed out. f Includes 2 animals without record of examination for original IBT report. Diagnoses for other 5 animals do not appear in records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter reports. Q * Includes one animal with both diagnoses. Hepatoma is 1 of 2 crossed out (superscript f). ^ h Includes 3 animals without record of examination for original IBT report. Diagnosis for other animal does not appear in . records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter ^* reports. u' HARTOLDMON0023367 bVOBbU WJi U IX X I B I T # 41 7 HARTOLDMON0023368 SnJudiiiaL B 1 O T E S T ^aJtey\alyu*i, Jtvt. REPORT TO ' MONSANTO COMPANY . TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH ARCCLOR 1254 - IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1975 * IBT NO. 641 - 06672 L Introductioa . At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity^ study of Aroclor 1254 in rats (IBT No. 622-07Z98) were processed into H St E stained sections and evaluated by light microscopy. The fallowing report presents the results of this study. . DEPOSITION | EXHIBIT | tje\inv#fa-*t1 `s-yv'ti uxc. SCM 058166 HARTOLDMON0023369 Jnduiixlal B I O T E S T }rtc. 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated November 12, 1971. There were six hepatomas detected among six of the animals at the highest treatment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. One liver tumor (a hepatoma) previously reported . in a T-II animal (No. 445) was re-classified as nodular hyperplasia. The . ' other treatment-related lesions reported are regarded as degenerative or . ' hyperplastic in nature and they are morphologic manifestations of an adaptive ' . - response of the liver ascribed to biotransformation of the test material. The . Hatter-lesions wie confined primarily to lest animals of the 12 and 24 month sacrifices and they were dose-related in incidence and severity. * f?r:: In conclusion, Aroclor 1254 does not appear to be carcinogenic in rats ; fed. for ;two years at levels up to and including 100 ppm. . Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. * Report Prepared and Reviewed by: D. E. Gordon, D.V. M. ,Ph. D Section Head, Pathology Report Approved by: M. L. Kepli/ger, Jpb, D; Manager, Toxicology HARTOLDMON0023370 JntLulxlai B l O - T 6 5 T IBT No. 622-07298 Monsanto 3no. 3 I have examined additional sections of liver from rats of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated my findings for Arodor 1254 on the following pages: * There is evidence of a chemical effect on the liver which is most pronounced at time of the 12-Month and 24-Month sacrifice. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoms. The hypertrophic cells contain large amounts of light staining cytoplasm which is probably rich in v glycogen and endoplasmic reticulum. In some cases ring shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen in the cytoplasm of these cells. . The hyperplastic nodules occupied larger areas and often compressed surrounding cells of the normal liver parenchyma. They also contained the same hypertrophic cells. ' ' '' * * Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence .of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study. SCH 058168 000 *vb HARTOLDMON0023371 jndultfiJsil B I O * T E S T 2aItaMlL>\U4, 3*C. 4 With. exception of the vacuolar changes in the cytoplasm of hepatocytes, the more severe hepatocellular alterations were confined to animals of the 24Month Sacrifice. (i/o/lel &Ccd&j Ward R. Richter, D. V. M., M. S. Dipl., Am. College Vet. Path. SCH 058169 O'lQ: .^7 HARTOLDMON0023372 5 ' Lesion Primary Liver Lesions - Aroclor 1254 3 Month Sacrifice - Rata CroHio TI th Tin Control Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cholangio -Hepatoma Hepatocellular Necrosis 3/10 0/10 0/10 0/10 0/10 0/10 1/10 3/10 0/10 0/10 0/10 0/10 0/10 1/10 1/10 1/10 0/10 0/10 0/10 0/10 0/10 2/10 ...... 0/10 0/10 0/10 0/10 0/10 0/10 SC* GOO 23 \ HARTOLDMON0023373 6 Primary Liver Lesions - Aroclor 1254 6 Month Sacrifice Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Duetular Hyperplasia Cholangio-Hepatoma Hepatocellular Necrosis TI 0/10 0/10 0/10 0/10 0/10 . 0/10 2/10 th 0/10 0/10 0/10 0/10 0/10 0/10 .0/10 Grouo Tin 1/10 4/10 0/10 0/10 0/10 0/10 1/10 Control 1/10 0/10 0/10 0/10 0/100/10 1/10 q58 sew /I 0^0 HARTOLDMON0023374 7 Primary Liver Lesions - Aroclor 1254 12 Month Sacrifice - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cho langio -Hepatoma Hepatocellular Necrosis H 1/10 0/10 o/io 0/10 1/10 0/10 0/10 th 4/10 4/10 0/10 0/10 - . 1/10 0/10 0/10 Grouo Tin Control 1/10 1/10 2/10 0/10 0/10 0/10 0/10 0/10 0/10 1/10 0/10 0/10 1/10 0/10 sc* 058172 000 ;o HARTOLDMON0023375 8 Primary Liver Lesions - Arocior 1254 Terminal Sacrifice - Hats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cholangio-Hepatoma Hepatocellular Necrosis TI 8/31 3/31 0/31 0/31 6/31 0/31 3/31 TII 10/26 5/26 3/26 0/26 3/26 . 0/26 1/26 Croup TUI 13/27 13/27 13/27 4/27 4/27 2/27 2/27 Control 1/23 0/23 1/23 0/23 5/23 0/23 1/23 SCH 58113 00Q'.:.3l HARTOLDMON0023376 * \ V^V- J..WU T a b u la tio n ot In d iv id u a l L iv e r Lesions in Rata ' * . .. - ----- ----- ,, tn *- s ( OH g B *2-- na Dose Level uoo ttiui . . . **odd wtia : :g , : 22~ wi 0 . U0Ol(UU0ttUIlU'lIUUIUIlUOIUI<UINOIUm0IU0h<IUJhI<0Ui<->I O<OOO0IO<O<0OI0IO< 1 aOO<<uO0Ou<0Ow<00IOu<<0uO<<MwOIOIuVOIUu0IIOuU<I sssshissssg : : : :hj : : : --> tHn 0ap. rv0 Ba3 + + i '. + + + ++ Cytoplasmic vacuolation of hcpatocytes ++ + Focal necrosis of hcpatocytes . Focal lymphoid infiltrations .. Focal hypertrophy of hcpatocytes Nodular hyperplasia of hcpatocytes L iv e r Lesions ... Focal bile duct hyperplasia '. '' Hupatoma ' %. Cholanglo-hepatoma i i SCM 1 000 Ov/l CD T f ,. ....... Squamous Cell Carcinoma, metastatic Mammary tumor, metastatic lie pat it is /Ci r rims i 3 1 HARTOLDMON0023377 6 AHUULUK - 1<134 Tabulation of Individual Liver Lesions in Rats acrifica 1 erval Dose Level - Anima No. and Sex * oo <u3 ^ >2 2 2. o .92) 2u cU 2 O *c. % a oc O ~a* ad 12 Ss* =w --_ C** iayd "uoid b h. Liver Lesions o 2* "5.2 ou >y -a wo oa iad. x>. xo yid o hi id _ 22 2-* oa. o --5b* ioas id -- l o. Z u id -? oo 3| oiyd hi cd 5 a 9 X d E 990Q. X1 C o U 61 vtonth B-I lppm 921M 922" 923" 924" 925" 936F 937" 938" 939" 940" n 2oa o > U B-n 950M lOppm 951" 952" 953" 954" 966F 967" 968" 969" 970" B-in 981M lOOppm 982" 983" 984" 985" 996F 997" 998" 999" 1000" r. + + + + + + SCM 058175 Squamous C ell C arcinom a, m etastatic iu |r*n HARTOLDMON0023378 \ i i SCM i - o \j\ 00 -4 ---------- O' . ... . .. -. **'*>#' s if O tjj S0 -B OiOM'IIMn^UMi. :ss:N|:s!<^ t - +. Sw . 9 --M a y OkUIUIUIUIUIUIUIUIW 040)>40'UlitwUNM . , 040>|IMII4kUMM + + t-t . + ++ . > (n P 2! E. x o. P 3* Cytoplasmic vacuolation of hepatocytes Focal necrosis of hepatocytes Focal lymphoid infiltrations Dose Level + ++ + + + . Focal hypertrophy of 1 hepatocytes Nodular hyperplasia * of hopatocytes Focal bile duct . `+ . + 1 hyperplasia 4 Hepatoma \ Cholanglo-liepatoma 4. . . . Squamous Ceil 1 Carcinoma, metastatic Mammary tumor, ' metastatic 1 HARTOLDMON0023379 ( i I S a c rific e I aterval i AROCLOR - l^s-t T a b u la tio n of In d ivid u a l L iv e r Lesions in Rats L iv e r Lesions o^orr HARTOLDMON0023380 ia o u u tio n oi in a iv ia u ii L tv e r Lesions in Rats L iv e r Lesions * ........ X ' . ___ . .. N ______ ______ g .* S & ____ ; * ____b* w -1 7h*P^SS-- nnH Dose Level i - 1R i^^^'J'J'10'0'U1U|>1'>UiUm0004)IMMNmhhh OMllUO>HO^UNOO>NHO^OUIUNh>>10'U<OOtIUI > W Df o m X P- " D P t ++ + +t Cytoplasmic vacuolation of hepatocytes + Focal necrosis of hepatocytes Focal lymphoid Infiltrations + ++ ^j- Focal hypertrophy of hepatocytes * *d*d Nodular hyperplasia . of hepatocytes * X ' # Focal bile duct +. hyparplasla Hepatoma i i i i i I i i i SCM o\J\ co *CD . , . tjrj ., 1 Cholanglo-hepatoma * i Squamous Cell Carcinoma, metastatic Mammary tumor, metastatic HARTOLDMON0023381 HARTOLDMON0023382 A U U C M I K - I <54 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats = m ild in se ve rity = m oderate in se ve rity u ixx V \- I B I T i ii i HARTOLDMON0023383 OnSiUMud BIO-TEST Jjzlxyiafoues. 3m. 1810 FRONTAGE ROAO ,, NORTHBROOK. UllNOIS 60082 REPORT.TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS . MARCH 24, 1975 IBT NO. 641 - 06672 HARTOLDMON0023384 Jndxjlxial B ! O * T E S T ^aJn>\aia'u&i, Jnc. REPORT TO ' MONSANTO COMPANY 4 TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 -: _ IN ALBINO RATS HISTOPATHOLOGICA L EVALUATION OF ADDITIONAL LIVER SECTIONS T MARCH 24, _ 1975 IBT NO. 641 - 06672 . L Introduction . . - . At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity study of Aroclor 1260 in rats (IBT No. 622-07298) were processed into H It E stained sections and evaluated by light microscopy. The following report presents the results of this study. SCM 058181 000 o .0 0 HARTOLDMON0023385 fndnilMal B I O - T E S T JaJxyialo'Jmc. \ 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated .November 12, 1971. ' .There were seven hepatomas detected among seven of the animals at the highest treatment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. The other treatment-related lesions reported are regarded as degenerative or hyperplastic to nature and they are morphologic manifestations of an adaptive response of the liver associated with biotransformation of the test material. In general, the latter findings were confined primarily to test animals of the 6-, 12- and 24-Month Sacrifices, and they were dose- related in incidence and severity. . In conclusion, Aroclor 1260 does not appear to be carcinogenic in rats fed for two years at levels up to and including 100 ppm. . Respectfully submitted, . INDUSTRIAL BIO-TEST LABORATORIES, INC. Report Prepared and Reviewed by: D. E. Gordon, D.V.M. ,Ph. D. Section Head, Pathology Report Approved by: M. L. Kepling'er, Fh>. Manager, Toxicology SCM 058182 Q^O ;o HARTOLDMON0023386 jnduii/Ual B I O * T E 5 T $*<a. 3 IBT No. 622-07298 Monsanto I .have examined additional sections of liver from rats of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated - my findings for Aroclor 1260 on the following pages: There is evidence of a chemical effect on the liver which was most evident at time of the 12-Month and terminal (24-Month) sacrifices. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals, to hepatoma or cholangiohepatoma. The hypertrophic cells contain large amounts of light staining cytoplasm which is probably rich in glycogen and endoplasmic reticulum. In some cases, ring shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen in the cytoplasm of these cells. The hyperplastic nodules occupied larger areas and often compressed surrounding cells of the normal liver parenchyma. They also contained the same hypertrophic cells. Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence of metastasis, invasiveness, severe basophilia, mitoses or other evidence of anaplasia, these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these , tumors in this study. SCM 058183 000 41 HARTOLDMON0023387 jridtiiUlaL B I O T E 5 T Ja&Moiyuu. Jnc. 4 With exception of the vacuolar changes in the cytoplasm of hepatocytea, the more severe hepatocellular alterations were confined to animals of the 12 % and 24-Month Sacrifice periods. ' (Jcuiej'R _________ Ward R. Richter. D. V. M. , M. 3. Diplocnate. American College of Veterinary Pathologists so a5818" 000 * 1V ^ HARTOLDMON0023388 5 Primary Liver Lesions - Aroclor 1260 3 Month Sacrifice - Rats Lesion . - Group TI TII 'tiii ' Vacuolar Change 0/11 4/10 3/10 Focal Hypertrophy 0/11 0/10 2/10 Nodular Hyperplasia 0/11 0/10 0/10 Hepatoma . 0/11 0/.10 0/10 Ductular Hyperplasia 0/11 . 1/10 0/10 Cholangio -Hepatoma 0/11 0/10 0/10 Hepatocellular Necrosis 0/11 0/10 0/10 Control 2/10 0/10 0/10 0/10 0/10 0/10 0/10 GOO:*A3 HARTOLDMON0023389 6 Primary Liver Lesions - Aroclor 1260 --------Lesion 6 Month Sacrifice - Rats -- - Group - TI TII T1H Vacuolar Change 1/6 2/5 5/9 Focal Hypertrophy 0/6 0/5 3/9 Modular Hyperplasia 0/6 0/5 0/9 Hepatoma ' 0/6 0/5 0/9 Duetula r Hyperplasia 0/6 0/5 0/9 Cholangio-Hepatoma 0/6 0/5 0/9 Hepatocellular Necrosis 0/6 1/5 0/9 Control 1/10 0/100/10 0/10 . 0/10 0/10 1/10 SCM 058186 ooo:vu HARTOLDMON0023390 7 Primary Liver Lesions - Aroclor 1260 12 Month Sacrifice - Rats Lesion ' CrouD TI TQ Till Vacuolar Change 2/9 5/10 3/9 Focal Hypertrophy 0/9 0/10 4/9 Nodular Hyperplasia Hepatoma 0/9 0/10 1/9 0/9 o/io 0/9 Ductular Hyperplasia 1/9 0/10 2/9 Cholangio-Hepatoma 0/9 0/10 0/9 Hepatocellular Necrosis 0/9 0/10 0/9 Control -1/10 0/10 0/10 0/10 1/10 0/10 c/to scM 058187 ooo:'i5 HARTOLDMON0023391 8 Primary Liver Lesions - Aroclor 1260 Terminal Sacrifice - Rats Lesion * Group Vacuolar Change ' Focal Hypertrophy Nodular Hyperplasia Hepatoma TI 5/25 3/25 0/25 0/25 TE 6/23 10/23 9/23 0/23 tie 10/27 11/27 7/27 5/27 Ductular Hyperplasia 6/25 5/23 14/27 -Cholangio-Hepatoma 0/25 0/23 2/27 Hepatocellular Necrosis 4/25 2/23 6/27 Control 1/23 0/23 1/23 0/23 5/23 0/23 1/23 SC* 000' *6 HARTOLDMON0023392 C lio la a g io -h e p a to m a Tnlnil.iUort of Indi/idiul Liver Lesions in Knls ooo:`.;7 HARTOLDMON0023393 AROCLOR 1260 Tabulation of Individual Liver Lesions in Flats HARTOLDMON0023394 HARTOLDMON0023395 C ho la u g lo -h e p a to m a 0-1 .t/l/l) HARTOLDMON0023396 AROCLOR 1260 Tabulation of Individual Liver Lesions in Rats HART OLDMON0023397 AROCLOR 1260 Taliul.ntion of Individual Liver Lesions in Rais 14 C h o la n g io - hepatom a Grading System + = minimal in severity ++ = mild in severity . +++ = moderate in severity ++++ = marked in severity P = Present, no grade SCH 058190 ono: HARTOLDMON0023398 THE EPPLEY INSTITUTE for RESEARCH IN CANCER October 31, 1975 Paul L. Wright, Ph.D, Monsanto Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear Dr. Wright; Enclosed please find the report on Hlstopathologlcal Re-evaluation of Tissues from Female Sherman Rats Fed Arodor 1260. As mentioned in this report, there were about 20 animals with lesions, which on a morphological basis alone, could be interpreted as neoplastic. However, the definite malignant nature of these lesions cannot be proved in the presented - material because of the factors mentioned in the section entitled "discussion" in the enclosed report. . I am sorry fhr I was not able to contact you by telephone. I was very much tied uj> with several visitors last week. If you have any questions concerning the report, please contact me. Sincerely, P, Pour, M.D* Professor TMj UfllMlllly * NlWIlU MMkll SCM 058195 IM O*--y Ahm, Omtiu,Naontfca OIOS ooo;,^ HARTOLDMONOQ23399 UJXX I ii \ I B I T #, I I HARTOLDMON0023400 Srt/SbJMud BI O - TEST J^mciyueS, Snc. 1610 FRONTACe ROAO NORTHBROOK. ILLINOIS 60062 . REPORT TO . ` MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1242 IN ALBINO RATS ' % HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS . MARCH 24, 1975 " IBT NO. 641 - 96672 DEPOSITION 1 ; EXHIBIT I IJ SCM 0581^9 ooa:*.5-i HARTOLDMON0023401 )tuLui\iai B I O T E S T jZabyial&utt, Jne. ' REPORT TO * MONSANTO COMPANY . TWO - YEAR CHRONIC ORALTOXICITY STUDY WITH AROCLOR 1242 ' *-----IN ALBINO RATS............. -- - HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS "TV ' ~ T_yMARCH _24, ~197S v ~ " IBT NO. 641 - 06672 ' . - L Introduction ' * ' :" ' At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity study of Arocior 1242 in rats (IBT No. 622*07298) were processed into H & E stained sections and evaluated by light microscopy. The following report presents the results of this study. SCM 058150 HARTOLDMON0023402 B I O * T E S T JaixHal&uM*, Stic. 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated November 12, 1971. There were three hepatomas detected among three of the animals at the highest treat ment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. The other treatment-related lesions reported are regarded as _ degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotransformation of the test material. In general, the latter findings were confined primarily to test # of the final sacrifice and they were dose-related in incidence and severity. In conclusion, Arodor 1242 does not appear to be carcinogenic in rats fed for two years at levels up to and including 100 ppm. Report Prepared and Reviewed by: D. E. Cordon, D.V.M., Ph.D. Section Head, Pathology Report Approved by: Id Manager, a oncology SCM 058151 HARTOLDMON0023403 jruLuUlal B I O - T E S T <ab>\alo^Ui, jtte. IBT No. 622-07298 Monsanto . 3 I ha/e examined additional sections of liver from rata of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated my findings for Aroclor 1242 on the following pages: _ I` There is evidence of a chemical effect on the liver which is most pronounced at time of the terminal (24-Month) sacrifice. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, * and in a few animals to hepatoma or cholangiohepatoma. The hypertrophic cells contain large volumes of light staining cytoplasm which is probably rich in glycogen and endoplasmic reticulum. In some sections, ring-shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen in the cytoplasm of these cells. ' , The hyperplastic nodules occupied larger areas and often compressed surrounding cells of the normal liver parenchyma. These nodules also contained the same hypertrophic cells. Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia, these are benign tumors rather than malignant tumors (carcinomas). There was no evidence of metastasis or invasiveness of these tumors in this study. SCM 058l 000:^7 HARTOLDMON0023404 jiduli'Ual B I O * T E S T JaJmMiloAUt, Jxc. 4 With exception of the vacu^ar changes in the cytoplasm of hepatocytes, the more severe hepatocellular alterations were confined to 24-Month Sacrifice. * I of the CJcuial WardR. Richter, D.V.M..M.S. Dipl., Am. College Vet. Path. SCM 058153 <no:*.53 HARTOLDMON0023405 5 Primary Liver Lesions - Aroclor 1242 3 Month Sacrifice - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Duetolar Hyperplasia Cholangio- Hepatoma Hepatocellular Necrosis TI 0/3 0/3 0/8 0/8 0/3 0/8 1/8 i\ Croup th Tin 2/10 1/9 0/10 0/10 0/10 0/9 0/9 6/9 0/10" . 0/10 1/9 0/9 0/10 \ 0/9 Control 2/10 0/10 0/10 0/10 . 0/10 . 0/10 0/10 SCH 05815* qoq:' r-n HARTOLDMON0023406 Primary Liver Lesions - Aroclor 1242 6 Month Sacrifice - Rats Lesion --------- ' TI Vacuolar Change 0/10 Focal Hypertrophy 0/10 Nodular Hyperplasia 0/10 Hepatoma 0/10 Ductular Hyperplasia ' 0/10 Cholangio-Hepatoma 0/10 Hepatocellular Necrosis 1/10 TH 0/8 0/8 0/8 0/8 0/8 0/8 0/8- Grout} TIH 0/3 0/9 . 0/9 0/9 0/9 0/9 0/9 . 6 Control 1/10 0/10 0/10 0/10 0/10 0/10 1/10 SCM 058155 ooo:.go HARTOLDMON0023407 7 Primary Liver Lesions - Aroclor 1242 t i l. Lesion 12 Month Sacrifice - Rats Group Vacuolar Change TI 2/10 xn ' 4/10 tm % 0/9 Control 1/10 Focal Hypertrophy 0/10 0/10 1/9 0/10 Nodular Hyperplasia 0/10 0A10 0/9 0/10 Hepatoma - 0/10 0/10 0/9 0/10 Duetula r Hyperplasia 0/10 1/10 0/9 1/10 Cholangio -Hepatoma 0/10 0/10 0/9 . 0/10 Hepatocellular Necrosis 0/10 0/10 0/10 0/10 i i i sen 058156 OOO.'Gl HARTOLDMON0023408 Primary Liver Lesions - Aroclor 1242 Terminal Sacrifice Rats Lesion ' Vacuolar Change _ Focal Hypertrophy : Nodular Hyperplasia ' Hepatoma . Ductular Hyperplasia * Cholangio-Hepatoma Hepatocellular Necrosis TI 7/31 2/31 0/31 0/31 3/31 0/31 1/31 Croup TH . Tin 8/30 . 9/20 3/30 8/20 2/30 8/20 0/30 2/20 3/30 .0/30 3/20 . * 1/20 . 1/30 0/20 8 Control 1/23 0/23 1/230/23 5/23 0/23 1/23 SCH 058157 0OQ:\C2 HARTOLDMON0023409 * ' ^ "* ' '* .................................. " / .* *- * * ' Ut ? l i* F J? s ^3U nA~H Dose Level C -I lp p m 1 c-u lO ppm 1 i 1 1 c-m lO O p p c i Con tro l ! A R O C L O R - 1242 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats o >oai<4 0|0<cA->i s : : :"l : : : :^ ++ 0'Jfl''aa,'J?' $ , o<aa-4 0o<aa^o! ++ o!oa-!S^o) sssslirS + X 1O *JL_ Cytoplasmic vacuolation of heoatocvles Focal necrosis of liepatocytes 3 Focal lymphoid Infiltrations * * * + Focal hypertrophy of hepatocyles Nodular hypurplasla f of hepatocytes A< Focal bile duct hypurplasla f-s Aa O a u Hepatoma o\j\ 00 \J\ , a> . . * Cholanglo-hepatoma teticulum Cell Sarcoma metastatic Mammary tumor, ' metastatic i i Tulani;ectasia. focal 1 HARTOLDMON0023410 1 1 1 SCH i AROCLOR . 1242 Tabulation of Individual Liv,r L^ions in Rat, Tclan^ectayia, focal ,6 Month C-I lppm ' 1011M 1012" 1013" 1014" 1015" 1026F 1027" 1028" 1029" 1030" c-n 1041M lOppm 1042" 1043" 1045" ' 1 J 1056F 1057" 1058" 1059" -| c-m 107IM 00ppm.l072" 1073" 1075" 1086F .1087" 1088" 1089" 1090" Control 801M 802" 803" 804" 805" 816F 817" 818" 819" 820" SCM 058159 II ono/.ci HARTOLDMON0023411 n o OarH*, 4>Qo-aotuiitoMfr-*. nl 0 %A ^0 %0 %0 0 %0 0 '2<5*i(5^WNe; Ua MJiL, >O=04=0)0O0=0O0=>04100O0=^0U0-0I|0^=0U00-N0^m0 CD _ O_ v-O 0-0- , (h Ul z od & n a Ar< oOM AA UNm CA/t A x a. o * > tJ 3 A (Cytoplftsmlc vacuotatlon of hcpatocylc^ Focal necrosla of hepatocytea Focal lymphoid infiltrations HA cr C_ a" r-r H* o 0 Focal hypurtrophy of hepatocytea Nodular hyperplaaia of hepatocytea Focal kilo duct hyporplaala > S3 C o r c :=> <1 HnA N1- r o3u Hepatoma JO A </> o r3>C o oUl CD r-* a* VI o Cliolanglo -hepatoma eticulum Coll Sarcom, me ta static Mammary tumor, metastatic Telangcctasia, focal , HARTOLDMON0023412 Tabulation of Individual Liver Lesions in Rats Sacrifice Interval '4 Month Dose Level [Anima No. and Sex 2 oo u < >tj. 2U (a4 91 -aS. o -ow >% u o2 U sU c* o4u t *3 c 0 <44 MW i3 CL ** = c *oo3 & C-I lppm 564M 566" 591" 567" .569" 572" 576" 580" 588" 589" 595" 60 IF 604" 60S" 582" 583" 607" 608" 609" 613" 621" 627" 628" 631" 634" 635" 606" 610" 620" 626" 633" f + Liver Lesions e > a.2 2 5 o >. a - **** O0 Vau aa>. a -= - *o u 0?.0-J O, w o= 4ou C<4 3* o Z mm o it -l 9O -- q_ f i* nou h ao. 4 M4o a. 0 j: 1 o "e3* 4 O 12 owu 4 (I VJ w-- <t u3 u0 o cd 5 if 3 a3 >.2 *A4 V sow Q58161 II ooorca HARTOLDMONOQ23413 ` to O " ''" o \J\ OD r-* O N) A R O C L o n - 1242 T a b u la tio n of In d ivid u a l L iv e r Lesions in Rats P P . * * 1 1 * . S HJ Hf~4 B a40^0'0^a4*>)ai4"4^1^j0^<h0^0^0^0^00'0^0^0'(h0^(h0'0^(h(h(h0' O vO *09 Q)HHOOOO^)\00090)0)Qa4(Aiki^i^^i(ki^i^(houlUi NmUNN^M^JUO^IOiO Ot^)4(hUHOChMii^UHO"4N*4Ul *% +1 + + ' T+ . ++ . ++ + .+ '' j 1 ; 1 .. * h- O <S N *Hn ` -n i 1-A oO ! a< nt ! *-- . > i %M"PaK. ?^ah23. . Cytoplasmic vacuolation of hepatocytes Focal necrosis of : hepatocytes Focal lymphoid i infiltrations + ++ i Focal hypertrophy of | hepatocytes tl " * + t+ Nodular hyperplasia 'of hepalocytus 1 Focal bllu duct 1 - hyperplasia Hepatoma 1% rH<A* *1 ST u oH* D Cholanglo-he patoma .* * Reticulum Cell Sarcoma metastatic , *d Mammary tumor, metastatic #l*i: l.l ui*i*r | s i.t , f r. 1 HARTOLDMON0023414 Cytoplasm ic vacuolation of hepatocytes Focal necrosis of hepatocytes Focal lym phoid Infiltrations Focal hypertrophy of hepatocytes C holanglo-hepatom a AIIOC LOR - 1242 Tabulation of Individual Liver Lesions in Rats Sacrifice i Interval Dose Level Anima No. and Sex r 1 ~j 4 Month 1r. 1 1 1 * r c-ni 740M lOOppm 722" 724" +++ 744" 746" 748" ++ 768F 771" + 77S" Ext" +++ 733" 765" + 766" ++ 767" 776" 773" 786" +++ 789" 793" H+++ Ext" ++ ' |1. . ( , r1. 1. Control 8M 11" 12" 21" 23" 33" 34" 35" Ext" 46F 47" 49" 51" 52" 56" 59" . 62" Liver Lesions .a a a|a. > 0c. o --2* ae. ue a-- 10 o Z Jj* e<sJ Q --O 9C. -3 2* oao fa a o . <C4. ------ -- -- . . . ........... P + P + 'P ++ P . +++ T + P. + +P +P +P p p +' . + +' 1 14 aC SCM 058163 I!I HARTOLDMON0023415 l> ^ n o 3C o Oin CD -- O -r Grading S yste m ' + = M inim *J in severity ++ * M ild in se ve rity +++ a M oderate in se ve rity ++++ a M arked in se ve rity P a P re s e n t, no grade .............................. 1 o 5 tr 1 -- ~ ' ' ( D rlv f*t1 <*$->* fHf*t* Dose A nim a Level No. and Sex C ontrol til 4 *>l O' O' O' |l M O O' 1^ u tt z z a z hj Cytoplasmic vacuolation + of hepatocytes Focal necrosis of + hepalocytos Focal lymphoid Infiltrations 1 Focal hypertrophy of I licpatocyles Nodular hyperplasia *d of hopalocytus Focal bilo duct > hyperplasia Hepatoma \ 1 Cholanglo-hepatoma L iv e r Lesions Reticulum Cell Sarcoma - ' metastatic Mammary tumor, ' metastatic rP<: h in*i*r t*i 4i r* f HARTOLDMON0023416 A R 0 C L 0 R - 1242 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats 9tvdujJAj&L BIO -TEST 1610 FRONTAG8 ROAO NOTHS0O<. ILLINOIS 60043 Snc. REPORT TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH . AROCLOR 1254 IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1S75 IBT NO. 641 - 0667? I i 0 nClPO HARTOLDMON0023417 ix m i I HARTOLDMON0023418 i ( I io.t*t Jahmta**. 3m." y |M MOHTAOa >0*0 221 ' Moamaaooa. iujno* <mi UMiT TO MONSANTO COMPANY TWO . TEAS CHEOTOC OftAL TOOdCITT STOUT WITH AE0CL0H US4 XN ALBINO EATS H3TOPATHOLOCKAL EVALUATION or AOcmcNAL uvxa sections MAACH 14, UTS I8T NO. 441 . 04472 ^ DEPOSITION EXHIBIT |cVHcrsfcrts44 1 s-an-ti <*>*4 i HARTOLDMON0023419 Maitmi *10-1111 fnhwttiiti IM MONTaO* 00*0 NWTNMOM.IUlHOtl t*MI kfaitk 14, 1971 JW 222 Georgo J. Lootaakaa, P1D. MoowMa Conftif 100 N. Uadborfh Bird. St. LmIi, Miaaoorl iJIM . Dear Or. Leeiaakae: la: ST No. Ml-OMTt Rlatsfatk*l>|lat Evaloatiaa W A41HmiI Ll<r Soctlaea frma lata o< a Two . Taar Chroaic Oral Taelely Stodr at Aroclor 1M4 Wo aro wbnilHl| herewith nr laboratory report dated March M, ITM: prepared U coaoectlaa with the aboro ready. Vary troly yeera. J. C. Calaadra Treiidoal J HARTOLDMON0023420 ,h+ml tlO-TIST 223 befoetto MONSANTO COMPANY TWO - TEAS CHSOmC CEAL TOBOCITT moTvasaoetos um at ALBINO BATS KBTOPATHOLOGJCAL EVALUATION OF ADDITIONAL LIVES SECTIONS MASCH 1VTS ' 1ST NO. Ml - 0M72 L lti4<ttlam At tbs milt of Dr. LtTiutai of tba Hamit Ctafur, iMUIhiI Mctioaa of Um from t two - yttr ekntlc trtl loUily rtarfp of Aroclor U?J la rata (I2T St. 4U -07IM) waro proeataof lata Hkl ataiaaf aactlaaa aad mhttN bp Ufbt microacopp. Tba hllwln report praaaata tba reanita of this ttadp. HARTOLDMON0023421 Uni (m Ms U art tthr >lfin< illp fata Mt ptirt--ly wpwal la ariflaal rspart 4W4 Rmartf U, ltd. Baaaiai. Mr* ^r* rts Mfi Knr.MMH 4s*to4 mwmm% rta rt M *slant* *l M U|tert tmrtM 1ml flM Ml rt M IHlirt kcriflM aMcfc nn art prsrloasly Npmrt^A* rtfcar llilrt*l-nli*l lastoas Wfrt4 tn i*fmra*4~V~ 4i|tMnM av VypstylasM la aatasa *a4 My wa rMiytil*|lf aaaaifsrtatlea* rt u *4ayttrs rsspaaa* rt M lira* mmSm! at* Mamiln imiIw rf M rt saariaL TV* Van laatoa vara rwflni< yrlauOr la tart aateaals oi il M Kaat 14 aart sa rifles* aa4 My n 4s**-r*lato4 la IsrUaan t*4 *rarity. la r*a*toslna. Aml*r 1234 ayyaara ta Va *Uhiiy aawl|rt( u Ural* rt 1M ppsa *V*a M etaMutf la M 4tot far M years. Ratpactfally nkalM QCOOSTUAL UO-TOT LABORATORIES, OK. HARTOLDMON0023422 BIO-TItT ibt k*. tu-mn ] 225 tU-tTiW aa* haw* w-nh>M4 >Mi tw w1l>w to WHtminU. tbm tohalato* ny flatfiaga tw AimIn US4 as Ikt taltowiag mil. TWn to nUni* to t tkMtal tBM *a to* ttm which u scat proaaeatai it ttoM to to* jfr-ltotoh ui MM*sth sacrifice. Thto ceestsc* to hapamratlalar iltmUH h|l--In m l*c*l hfpaatraphy which pnfniM* to ctolcr hyperplasia. tmd to lav utatli to hapatoao* *r rhalaaglab*patents. Th* hppailiaphla e*U* fM*l Iccjc umm ai Ugh* atolaiag cyteplaam which la proi ilf rich to glycegaa ui niepUimit rettrehwa to aac* *--a iii| atop** *teuton*, prahahty reprenadag wharla * prelUaratH* aaheplaamic ntinho, were *m* to to* cftoptoaa at to*** Mils. Th* hyp*rplactic nSale a aac^is* larger arcaa tai *fa*c cempresseC nmuhii call* to to* aancl Unr psrsechym*. They *U* c*at*to*4 the asm* hppailiaphit cell*. Than toatoa* which I claasiAe* a* toperm* a *r chelaagle-hepeaemaa were larger a*Ma* which ahaweg *wt4*aa* to caaftaaac* *r acne Tartetlsa la call sis*, ahap*. atolaiag *r a towtotor *r a Aaaamiatmia patters to giwwth. to toa ahaaace tosaatoataala, towastewaeaa, ****** toaaphlUa. aaitaaaa. *r ether awtoaaaa to aaapiaato; 1 ch*M to aaU these toaigs teaser* rathe* thaa maU**eai (carciawtoas). There wsa as avUaaca to mataatoaia er toessleeaea* to toss* HARTOLDMON0023423 Wttfc imn W>miUiIt tlfnltm am tMk (mtIOm. f ,U (*!- W. W*r4 ft. fticfcter, D.T.M..U.S. MfL, Am. C*U*t* T,t- fMk I "CT'1 "7*^ 000;,7G HARTOLDMON0023424 227 Primary Lhw Uiint Amlw llS4 ) Mmk OaarMWa lilt l' Urt-- Vmiiir r>m rt s/io T8 3/10 9tflt TZS ^w * Camral 4K t/io U 1/10 Toetl If>yaUoyhy MaOalir HyyaryUaU 0/10 0/10 0/10 0/10 B^mma Dactalar HyptryUila 0/10 0/10 0/10 0/10 Chalaafto -Hayatama Hayatocallolar Nacraala 1/10 1/10 0/10 0/10 000 0 >>* .V 4 / HARTOLDMON0023425 4 '* a.-*?; A' -, . 1 d*-*'-. tC- v Maur Uvar Laai-- - AmtatUM " 1 '* Iktalk ImiUIm . )Uts J'". - Laaioa Vuwlu Cbu|t Focal Hypertrophy Nodalar HyparpUri* BayatMBa Daenlu HyparpUaia Qtolaafio-Hapatotaa Hapotocollolar Nocroola TI 0/10 0/10 0/10 0/10 0/10 0/10 1/10 Tn 0/10 0/10 0/10 0/10 0/10 0/10 0/10 vm v 1/10 * Coatrol 1/10 4/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 1/10 *5% 1/10 HARTOLDMON0023426  Pilury live* Uia - irocUr 1254 14 UnIh OaarlOca - UM Lio Yuiu rv r*c*l H>yiilioyh; No<tei&r RyyarpluU Hifitgot tacdu Hyparpteala Cholu|ia-ifap(taB* Hptoc*UaL*r Macroala a 1/10 0/10 0/ld 0/10 1/10 0/10 0/10 in 4/10 4/10 0/10 0/10 1/10 0/10 0/10 V Of*- TO *>?. CMW 1/10 e/ 1/10 2/io 0/10 0/10 0/10 0/10 0/10 0/10 1/10 0/10 0/10 1/10 0/yo 0/1C T * a HARTOLDMON0023427  Llm lull * AncIm 1254 TamOaal aarlflaa lu* Latlaa Vtnoku Cbi| Total Hypartaoptor Nodular Hyparplaaia Hapatama Dactolar Hyparplaaia Cfcolaflo -Hapatama Hapatocallular Naaroala 3 '* >3 0 * T1 Til l?fe **. (* * 4 4/31 VvVd 10/24 -/itV. I3 v.H S/Sl />.?/ S/24 y4,r*-( 0/31 % i/24 ; -V . W* 1J/2T ^Ail/23 0/31 0/24 V 4/21 >-.i^4e/25 4/31 vv VJ 3/24 v.V 4/21 S/23 0/31 0/24 tr z/n Vi-lJo/u 3jt/-3. 1 vsy*< 1 >* na. * Lr 1/24 *u. VL< 2/21 " * " 4 , ^ M'-- fr HARTOLDMON0023428 miMtoilc TC?mni7CIFfOT7 Sacrlfic* UMml Dm* L**ai ABOCLOft US4 M Lhtr LM*m la Uwmt Uoteao H*. Md r !i ft 11 it ii li ii i 1 Mootfc N.' *. * B-I iu ur Ml" Mr )40" JHT irr* " m** 400" B-n 434M \ lOypm 4J1" 4)1" 4*4" 440" 47tT 477" 47" 474" 400" B-m SUM lOOffm S17" St" sir* U0" sisr SI7" >" nr 144" *** til aS HARTOLDMON0023429 Aiocua tm TiMiIIm W Urn UiUm la Bala 23-2 to ' *T' ooor.c2 & HARTOLDMON0023430 .1 HARTOLDMON0023431 jp- . -5 f'i* h" ^ 4j*s -V' - - -; 'T-i--fASssSAte'..*j*s- - -. ' ^. .-~ r-'>; ;.x\ -i.< ***; -w . - -* - - - .^V- -''% x "'V--'' ^'-s?'3^*^ .'>:'*'*.*. HARTOLDMON0023432 AROCLOA US4 13 HARTOLDMON0023433 lla|iaiiiita /C lrrh o ili AltOCLOft . 1*94 TtfcaUHoa al UItUwI Um UlUu la IUU 236 3*c rifle* l^arral Do** Aaima Ltvav Laalaaa Laval No. oad nSa* i! 21 ii ItV }! al Vaa* * h * li II i! 1 5J 1n * 2 i i i u ! i j T 3 0 1X U i 24 Month B-QX 4I4M rl+4 lOOppm 491" N ^ / 497" 499" * S07" 0 909" 910" 912" 443" r* 409" t* 9X2T *44 924" 9*4 V 927" *444 p,4/ t ** 941" Id 947" Eatl" >44 Ext2" *at3" 19+4 CsH" 9 Cst9" 934" 9*4 937" 1* 939" 44 441" Xal" Ea2" 9*4 ta3" -144 H4 (v) i**d ti p*" H Pi-* hp h P P ) 1 1 IfJ >1 * H* 1 l * "I 44 ip.1 t* ]r> 9* H * H Pi" [44 ir H Pi-' IP H P * w PIP1 H W P H |>444 H tt IP 41 P liP M-P >9*44 4 P M C*"' i l PH ! L rt*4 h l>4 CP' * * lev p * ! J W P 1- I H Pl-I r if- 1 2* ! iI ! i j H I I i Grmdlnt Svitaia ,i |a Mvaiitjr *> mild la aavarttjr ~h- modaraaa la aavarltr hh nurhid la aavaritjr P Praaaat. ao grad* Abaaat . HARTOLDMON0023434 U <AW tlO-TIST z 237 11. Summtrr 't -'- A In roost instance s,, treatment -related hi stops tholo. .1 findings la tha livsr from this re-ewlnatiae 414 cot 4illst slfniilcs; from that prttiouly reportsd In our e If ins 1 rsport dated NoTStnbsr 1Z( 1971, Hmnr, tbs's vsrs six bsalfa Unr musts 4otscta4 smoaf six of tha animals st ths hightat trsstxnsat level 1104 ppm) of tha 24-Month Sacrlfica which wars not previously - I. - '- l .- * ... . .J, ' ... reported. Tha othar treatment-related lesions reported a re regarded as ` degenerative or hyperplastic la nature acd they are roorpholofic manifestations of an adaptive response of tha liver associated sitk biotransformation of the test rraterial. Tha latter lesions wars confined primarily to test animals of the 14 and 24 month sacrifices and they were dose-related in .evidence and severity. In conclusion, Aroclor 1254 appears to be slightly tumorigsme at levels of 100 ppm wLea fed contlnously la the diet for two years. Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES. INC. Report Prapsred snd Reviewed by: D. E. Cordon. D. V. M., Pta. D. Section Head. Pathslofy Report Approved by: M. L. Keplt Manager, Toxical HARTOLDMON0023435 fr&atod Bio-Tisr SrtHtc&vu, JUc. 1*14 HONT14I *OAO - ,, NO*TM*fcae.-u.iMoi* mhi" - 188 REPORT TO MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITY' STUDY WITH AROCLOR 12*4 IN ALBINO RATS HSTOPATHOLOCICAL CYALCATI ON or ADDITIONAL LIVES SECTIONS MARCH 24. 1479 1ST NO. 44 . 06672 /' HARTOLDMON0023436 t frkntwl BIO TBT NOtTMMOPt, iUIMOII 189 C>ni J. Lovteakaa. ftJ). MO N. tgdbaryb Slrd. St. Lmu, WHiwrt 4J144 Dor Dr. Lortaskss: IU: IBT No. 041*04472 - Hlotopatfcological Zraluslhoi o2 AMlttaail Lirar Sorttatu Fna Rot* l 4 Too Tor OnaK Oral Toxicity Stadr at Aroclor UM. Wo aro NbaiWi| hvnrttk oar rrruod Ubnnlory rapart doSod Msrcb 24. WTJ; proporad la with Oa abort atlady. Vary truly yours. J. C. ralwidrs a _i HARTOLDMON0023437 IO.TIST JU s'a- 190 KEPOBT TO MONSANTO COMPANT TWO TEAS CH3GN1C ORAL TOXICITT STUDT WITH ABOCLOR 12 J4 IN ALBINO RATS KBTOPATHOLOGICAL EVALUATION OT ADDITIONAL LIVER SECTIONS MARCH 24. 147S IBT NO. 441 04472 L Introduction At tan n^Mtt of Dr. Lsotaskas of tkt Uuiutt Cwfur. additional sections of Uoer from a two - year chronic oral tosicity iMy of Aroclor 1254 la rata (IBT No. 4LI-07Z44) voro processed Into H 4 E itaiaad aoctioaa and evaluated by ILjht microscopy. The following report prv .ants An roaulta of this study. OOO'CO 4 HARTOLDMON0023438 IO.TIST 191 n. is tha Uvar iofIra i w wiHiHIi rili III rapartad in oar nrlglsol ropart dalad Rmab 12, lf?l. sis It mips livor tamorr 4stsctad ananp Us at As i Irvol (100 ppm) at 1 Ou livor tunnr (s hapatas) provtsusty nforOi in T-n IQhop I Ola. 445) ni ro-daastfiad as iidilg hyparplisii Tha ithar trsaMsnt-raialad laoiaaa roportad arc rspardad aa ihfillii or liipaiplanlc in mnn t nd thoy aro norphalopic nanlfaatatinna at an ailapWvi roapenaa of tho Uvar aiaikad la biatranafni nation at tha loot notarial. Tha Unsr laaiana wars rwiflnsd prtaaxily to taat animals at tha 11 and 24 aarriflraa and thay wars dsaa-ralaasd *n inriftonra and aovarily. In rwicluaian. Aroclar 1254 appaars bo slightly tunorlgooic at at 100 ppn wbaa (ad rmtinoaaly in Aa dial far two 7oars. Raspectfolly nhaUMd. INDUSTRIAL BO-TEST LABORATORIES. INC. Report Prepared and Raviowrod by: D. I. Cardan. D.V.M.. Ph D. action Hoad. Patfaalofy Roporl Approved by: id U. L. KapUnpor^Ph.D.7 Manager. Tanlralofy ' HARTOLDMON0023439 BIO.TUT tBT No. 422-072. _ J9Z ] I ktff mmliri i^dltlMil sections of lifu Inm rUi at My Msmbtr 422-0T29# ud bare n-nluM4 them (or evidence ti rsrrinngsassti { ban tabulated my Htdlip for Aroclnr 12S4 a the following yt|ti. Thnm U evidence of a chemical affaet oo tha liver which i moat pro nounced at rlma of tha 12 -Month aad 24-Month sacrifice. Thia consists of a hepatocellular alteration beginning aa focal hypertrophy which progmoot :a nodular hypo rplaala, aad la a faw aaimala to bapatoma or cholangiohspatoma. Tha hypertrophic calla cootala large amounta of tight ioiai| cytoplaam which la probably rich la glycogen aad eadoplaamlc rvtic-ulum. la some cases r-.ag shaped stroctaree, probably repro sooting wharla of proliferative endoplasmic roticuluin. wore saaa la tha cytoplaam of thata calls. Tha hyperplastic aodulas occupied larger areas aad often compressed surrounding calls of tha normal !' !* careachyma. They also contained the same hypertrophic eella. There laa lone which 1 claasUiad as hepatomas or cholaagto-hepatcmaa ware larger nodule# which showed ortdeoce of coafluoaca or soma variation la call star, shape, staining or a ductular or adaoomataua pattaru of g-owta. la tha abeeace af mataetaals, lavaslvenesa, sera re basophilia, mltnaes. or other arldanca of aaaplasla; t ehoaa to coll that# benign tumors rather than malignant (carcinomas). Tha re was so erideace of metastasis or Invasivetie as ef these tumors la thle study. . -- HARTOLDMON0023440 mor* Mnrt hayatacaHalar tltintloM war# mart--i la aalagats af tba 24llnath Sac rifle*. AiUtr Wax* *. ftlcktar, O.V.U..U.S. DlyL, Am. CaOaga Vat. hA, I1; V . . *a . w, 000:33* HARTOLDMON0023441 roc*l No4*la* Hypirpluil Ihptont DactfeU* Hyparplatla ChoUofto JWpatoma HtptlaciUiltr NicrMti 0/10 0/M 0/10 0/10 0/10 1/10 o/M 0/10 0/10 0/10 0/10 1/10 mo 0/10 0/10 0/10 0/10 0/10 o/io 0/10 0/10 0/10 0/10 0/10 1 HARTOLDMON0023442  195 t Primer? Um Udou - AroeLor U54 1 Month Sacrifice JUt* Vncnoler Cheng* foal Hypertrophy Hothiler Hyper?tea le Hopetome Docraler HyparpUoie Choleegio-Hopetome Hopetocolluler Necrool* TI 0/10 0/10 0/10 0/10 0/10 0/10 2/10 TH 0/10 0/10 0/10 0/10 0/10 0/10 0/10 TIB 1/10 4/10 0/10 0/10 0/10 0/10 1/10 Control 1/10 0/10 0/10 0- 10 o/'.o O'ta 1/10 HARTOLDMON0023443 7 196 i Primary Liver Liilou Aroclor 1254 12 Uoatfc Sacrifice lilt Laaloa B th Tm Control Vmtlu CUa|* 1/10 4/10 1/10 1/10 Tecal Hypertrophy 0/10 4/10 2/10 0/10 Nodular Hyporplaala o/io 0/10 0/10 0/10 Hapatoma 0/10 0/10 0/10 0/10 Ouetular HyparpUala 1/10 1/10 0/10 1/10 Cbolaaflo- Hapatoma 0/10 0/10 0/10 0/10 Hepatocellular Naeroaia 0/10 0/10 1/10 0/10 V. HARTOLDMON0023444 It7 Primary Lhtr Laaioaa - Aroclar 1254 Tarmiaal Sacrtflca Rau L*ties VtcooLar Toeal Hypartrophy 'TothUar Hyparplaata Hapatoma Sttutular Hyparplaata Cholanfto-Kapatoma Hapatocailular Nacraala T1 8/31 1/31 0/11 0/31 6/31 0/31 3/31 Til 10/26 3/26 1/26 0/26 3/26 . 0/26 1/26 Grow thi 11/27 13/2T 13/27 4/27 4/27 2/27 2/27 Ce. 1/21 3/23 '.<23 0/23 !/23 0/23 1/23 HARTOLDMON0023445 AHOCLOR . UM TabaUUo* oi badlvidaal Linr La*Ion* la Rata * 198 Sacrifice Lota ml ] Vtoath Do#* Laval Aaim* No. 1 and 2Saa a nh i! * "i si M S&'* 1 U u X u 'B-I lppan l J5 4M 157" 151" 15 9" IlM11 39tr 197" ! J" 199" 400" 9* s-n 41MI lOppm 417" 415" 419" 440" 476r 477" 475" l479 450" 1 B-EI IS16M lQOppm! 5 17" 511" 1519" 1520" |555r ! 5*7" 55 559" SoO" * * - 1 * 1! X Uvar Laalaoa If l 5J 1* * m m H 2& *i l ! 1 I i I . i i 1 j i i ti lj1 i I i ; i i > i i i i j i i ; i x X ilii'll J . t -A H ea - 12 is r4 1 5 ! I 5 5 r3 2 -1 HARTOLDMON0023446 a? o B- oW _____ ___f ^ <A >0 >0 A 9 : : :^ : : ? :V o^ 04-94-O)t`o^>^liPKJ<t^WWUNM^J0i . . : : :n : : ; : r 3" B ^0o^>w0o 1>uo W4 4U9- N4Ui N#t NU 4NN) N<>A : r :^ r IT? 2S -----v iHf 0?~ o ff |Tyioplanilc vacuuUllon of bopitocylPc Focal Mcroaia of hapalocytoi Focal lymphoid Infiltration* Focal hypertrophy of hopalocylaa Nodular tiyparplaala of hapatocytaa Focal hila duel fcyparpfaala acr eF. oo t n E* r* rZ F 9, lUpatoma * C he la oglo* hepatoma S<|uamoua Coll Caretnoma RMUplatir Mi mm* ry tumor, malaatalic |l*|taiii /Clrrlmala to to #* HARTOLDMON0023447 AROCLOR US4 TabaUtiaa o4 lattrUaal Uaar Laaioaa la lata u ML Sacrlilca Doaa |Anltna| Uvar Laatoaa Intaral Laval No. and * Sax ns 2J inl Itii *1 ill -1 9 V u H 0 Z li 5& a 8" Sm S 1 1 2 S sf Is l\1' H? P !5 i V '}y 3- !j Sr 12 Month B-I lppm 41M 42" 43" 4S" I \ 4br I i 47" I I * I 491 ' 50- I , B-a I SIM lQppm ; 52" I 53" | 34" 55" ! s&r j 57" I 5" I 5 9" | b0" B-m , lOOppm' ! bIM | o2" j b3" b4" 65" wr 67" 61" 69" 70" % HARTOLDMON0023448 HARTOLDMON0023449 llo |M liiiN /r Irrlim ii A ROC LOR . I2H Tabulation ol Individual Llvor Laalaaa la Rata Sacrtilcs Intarval Doaa Laval Animal JNo ^' Saa It si -1 u r I 24 Month B-S UlSU lOppm j 417" I4U" i 4It" j 423" j 426" j 427" ] 491" J 402" 403" I 40*" i 410" | 434" |4$or | 451" 1452" I 454" ! 459" J 443" 1 469" ; 470" ; 471" 473" : 443" '445" Ij 440" Llvor Laaioaa si u I Is tl = 11 i'i 51 4 kK si ia 9 Z It n P P l i * ? a J U 202 13 ft HARTOLDMON0023450 A HOC um . 144 TibaUllM ot lndtrWoal Ltnf Luioal la Ratt u 203 Sacrifice latarral Data Larat A alma No. i aad 3 Saa J a H Hh n *w il M iJ V1 kk u * fl 11 ii k Lira r Latloaa if 3 m * t 2L i i! i 8 k i ( 1 isii s', iz If e a it 2i Hi! 1i ? H4 1: 2 lu u 24 Month s-nx lOOpptn ' . 444U +4- 491" 49?" 499" JOT" 509" 510" 4 512" 44J" 449" 522P 524" 44 4 4* 44 52?" 4-4* 541" 54?" EatH *r Eatl'l EatJ" ** E*t4" Eit5" 5 36" 55?" 559" 4 441" bl" Ex2" 44 EsS" * 4p 4** p 4p p p * 4 * 4 4-i~P4 p p p p p *** p * pp p -* p *4 i p p* 1 ? p1 1 p Grtdlni Sanaa . minimal la Mftntr * nlU la taratitp nwltnti la torarity m marfead la Mrarity P Praaaat, as (rad* Abtaat I- HARTOLDMON0023451 * - ' .; -r- . -.''-vfr* v-n^.s^VtviVir'-r.3-/#if s'^*:*_ . '-yr/ 5J.\S/***X Z_ - ` i-.-t..-/..*;->--*T*-'. . y -* Jf.. 'is*.*" -. > i-'^xw-r ;,, -y . . ;, . - ; ' i' S'-V' ' " f i-i. > .- .Att ; * oofcfrtWyHl i--ft . . .-, l^lfO.TIIT jlOAfwMO--. IkUNOt* H HOQMtO MONSANTO COMPANY TWO YXA1 eittONje ORAL TOXICITY rrrov wtm AICCLOI M IN ALBDIO EATS KmONATNOLOOKAL EVALUATION or ADDITIONAL UVOt IICT10M MABCNIi. IfTI n IDT NO. Ml OUn HARTOLDMON0023452 ai arm. Hi-*un - mta^trtitiogini u Lfrrar Socflaoa fna Ui af a Twm - Taar Ckrak Oval Taaldty btadr ad Awriw W. Vt a* adbartMag >awltt --r'rarUad laboratory r^orl datod Mack M, ltl), fuptrod la eaaaacttaa akk At abort otady. Tayadritat. JCC/U HARTOLDMON0023453 4 WiUrf ftlO-TIST -',058 (XPCtT TO MONSANTO COMPACT TWO - TEAS CXBCNK OftAL TQXICITT STUDY WITH ABOCLOB USA IN ALBINO BATS H3TOPATHOLOGJCAL CVALCAT1CN or additional litzb sections VUICH 14. in 3T NO. Ml - 044T2 L at24actlaap At tfca roqorat a( Dr. Lavtaakaa al tha UntuM Compear. tAditiseal octloa* af lltrar (ram a tv* poor cknalc aral Mrtcliy itt^y at Araclar 1254 - la rata (ST N*. 622>97290 vara pracaaaod lata Hit mM Mcttooj ut rralaata4 by U|ht microacopp. Tba tollowlaj report praaaata dw raaalta it tfala tto4r> HARTOLDMON0023454 ^ t kkr ---- wara iwiflaai prlwarily la mi ataaU of fta U od4 14 sob* * MerifliM m4 flwy vm tefitod toi iTiikif< Md murliy. la mmMm, AwHg UM ifpw to bt akgfidy nawiptor at Mi i3*m, ir~ I*' **" ,twa U *-!' jj * v . . .. i 1M ppai otw U eaUmcIf la tha diarl lor Mm yaara. , KxnrnoAL no-nsT laxcbatowis. me. Xaport Pf>n4 ul Irrtm4 feyi D. X. QrH--. D. V.U-.Pk-D. Sactlaa Hm4. PatiuUfy Xapart Apyrml Vyi HARTOLDMON0023455 cHARTOLDMON0023456 HARTOLDMON0023457 HARTOLDMON0023458 i 083 Primary Unr Uitu - Aroclor 12S4 i Maa* Sacrlllaa - Kata La a ion V molar Chanja Tocal Hyportropby Nodular Hyparplaaia Hapatoma Dncraiar Hyparplaaia Cho Ua*lo-Ha patoma Hapatocallular Nacroala TI 0/10 0/10 0/10 0/10 0/10 0/10 2/10 TQ 0/10 0/10 O'lO 0/10 0/10 O'lO 0/10 2S!9L Tin 1/10 4/10 0>10 0/10 0/10 0/10 i/io Castral 1/10 0/10 0/10 0/10 0/10 0/10 1/10 i HARTOLDMON0023459 rtm cT / Mmit U*wr Uitaa - Atmlor US4 U" " - uu JOClSS Vmotu Chi|i Foul Hyyrtmyhy NoOvlar ftyyarpluU Rapatcma Dacmla* HyprpUii* CboUmfio -Hiptuat H>pttsclhUr NtertiU s 1/10 0/10 o/io 0/10 1/10 0/10 0/10 TH 4/10 4/10 0/10 0/10 1/10 0/10 0/10 nn. 1/10 2/10 0/10 0/10 0/10 0/10 1/10 014 : Caaml 1/10 0/10 0/10 0/10 ' 1/10 0/10 0/10 T HARTOLDMON0023460 -A> c Mwr Umt Urtw - ttXr Tiwrlr-* hotflc* ftatt r**i8i f i i 0/23 MwtaUr HyvT>Mto om /20 13/2T 1/23 B>ptnw 0/11 0/20 4/2T 0/23 DvctsUr Hyp**?***4* tm 3/20 4/27 3/23 / CfcoUnfto-B|W*-- Hapttocallate* Wril* 0/31 3/31 . 0/20 1/20 2/27 11 /2? 0/23 1/23 HARTOLDMON0023461 i 1 %\ fhif xm i u ml |CflofUinic vicmUUm * X q| Upitocyfl fw>l (MrMll t ____ hlfilKfllt Focal IfnifteM WllliUlMl Focal fcyyartroy Iky of hayatocyloc IMaUr Iqftrybila oI kaflHqrMi Focal Mia 4ocl byparylaala llayatoma Ckola<nlofcayI--' f *P I> is h i* jf I wV m . loonmi Coll ' Cowl--btioteHllc Mammary locnOr, malaalalla np^iTilta /Cirrkoala' i HARTOLDMON0023462 J 5acrLOce Interval JJfci ABOCLOB . 1*94 Tabetatlea ot Mhrlduel Liver Utltu la Kata Data Aalma Laval No. Llvar Laatom So * Jitl >t 1i si * 8. ji 14 m 1 li 3 !i 40 4 I* -V: ii i! mm 2 L 1 2 V 2 1u B-t 92IU 9Z2" 9*3" 9*4" 9*5" 934T 937" 93B" 939** 940'' B-Q lOppm 960M 951" 99*" 993" 954" 944T 947" 949" 949" 970" B-tS 99134 lOOppmj 992 993" 994" 995" 794T 999" 999" 11000" 067 2m 2 iJ5! jfc 10 :: i x It HARTOLDMON0023463 r \ cf J li 1" fa 1" f a? 1- *** n , ^ wf t * i tt t Cyafti--B<i Mlillw i VayiMytaa ' Focal awiatli of iay aarylaa Focal IfMfkaM lofUtratiooa r, " Focal fcyyHlOhy at fcopaaoay' a -1 ** Ma <ali.liyporylaaU a1 fcoyalocytaa Focal Mia M * r lyaiflaala . 1 m \ S V ' . '> . Kr*// i.''* jt ^ .b'?15>"&.. / ' % : 'SW ' V , i " 0 . l C|whgla4 |fo>aaO (luuaanooa (Soil ' Caraloowia. malaalalia Mjnimtri inmor intfUfUtU lk'|i4tllU/(!lrclioii * V HARTOLDMON0023464 T tta U tiM oI U ir M n tl L in t L a ile ii la t i l t A ^ M <0 O) *4 A i* * N K0 1 ' f" O 4 M 4^*49>^9>U'^aiNN9'wUiW^UiO^ 4 +x t i *' : ** * -1 S-Vl* -1? 3s *- r 3: n? 1 1 Cytoplaatrlc nctulillai of Lapatocytaa/'^ ' Focal naaroala of . UfaiaayHi' ' Focal lrmpM4 ..jj-w loflltratiaaa^" Fooal hypertrophy .01,^. hapatacytaa ^ t t t t Nodular hyparplaala ; of hapatocytaa v^di f *1 Focal blla *aa4- ^;',^' r hyperplaoU^;^. Se' llptonM * .i CbolaaflobapaM*)*^ , % . v A <%* .* lyMOtai Coll ' Carclanim.iinlOlataUc . . .<r`i Mammary tunauri _ .. malaalaCc ---------------------------------------------------------------------------------------------------- --------------------------------------------llapatltla/nirrhoala I HARTOLDMON0023465 AAOCLOR 1234 TabvUtim f Mlvidaal Llrar Laaioaa la lUta SacrUlea latarval V- Dot. Kn'ja* Laval No. IfSu a Si it a1 21 u i<* V e UmLyitu ii hii ii 1l -j ii i k I i ; 1 i 24 Month B-n 413M lOppm 41?" 413" 419H ! 423" 426" 42?" 491" 402" 403" 403" 410" 434" 430r 431" 432" 436" 439" 443" 449" 470" 471 473 443 443 P P 070 u a i i If i f* i: e i iI II I< HARTOLDMON0023466 AHOCLOR No. u4 Su 1 m ................................. ...................... * K o p 6 ................p 1 ! i | nV t 5 nm r? 3: ppptsssppppprjssssj^sssttsfc 3 3 [* 3 4m4N^UNO<*4>*4m <4 t+t{}**+ +*+ + Jt a {it i ti * o O'O'O dad, >o >o t d < *o *d *a n <d Cyloplaamlc vacuolallon ot Hapatocytaa Focal oacroala of hapalocylos Focal lymphoid Inflllrallona Focal hypertrophy of hapalocylos Nodular hyparplaala of hapalocylos Focal hlla duel hyparplaala llopalonta r 3 9 r m r 0 HK oar Va. 8 a l \ d *d U C holag16-ho pa1oma SqwmMi Coll CrclAemt#iMUilall Mammary tumor* mataalallc 11 |Mli(k/Ckrrhoali HARTOLDMON0023467 Laaioo Vacuolar Chaaga Focal Hypartrophy Nodular Hyparplaaia Hepatoma Ouctular Hyperpia ala C holanflo-Hepatoma llapatucallular Nacrosia o 072 Primary Uni Uriou - Aroclor 1254 Tai inlaal Sacrifice - Kata n 1? 1 1/11 S/2S S/Sl TO 12 Crap TD1 1 12 1 4/26 4/25 10/26 11/27* 1/2511/27 I/ll 2/2S 1/11 0/20 0/11 1/26 1/25 5/26 1/26 1/25 1/26 6/27 1/2511/27 1/27 9/2511/27 m 0/11 0/15 0/26 1/27 2/21 4/27 S/ll 6/20 6/11 4/2* 0/11 2/11 2/20 1/11 1/26 4/25 1/26 0/25 0/26 1/26 2/25 1/26 5/27 2/25 4/27 0/27 1/25 2/27 1/27 1/25 2/27 Control 12 1/21 0/24 0/23 0/24 1/21 0/24 0/21 0/24 5/21 6/24 0/21 0/24 1/21 0/24 1 - FI ret anlutioa oi ari|iaal slidae 2 - Ra-evaluation ci oH|iaal alldai 1 - Evaluation of aitiMUnaal tiaauo aacHaaa Thera nra 5 aaintala ia which tMa laaioo was not aaaa at Urn* of Iha urnad avihaaMna hat waa praam! on I he firat. 1 Similar to abova tavohrlag 4 aalmala. ` -- a u HARTOLDMON0023468  s 3 1 n HARTOLDMON0023469 uuu <1* HARTOLDMON0023470 HARTOLDMON0023471 HARTOLDMON0023472 t { E> xi H I! B| II HARTOLDMON0023473 ? 11990 m ON IS I %u>\ >* Hosvn SKOIXD3S (3ATI IVNOIUOaV JO NO IXYOTVA3 IVDIOOIOHXVdOXSnt S1V ONJS1V MI <* rm hotdosv hxu xanxs AXI3IXOX 1VSO DINOSHD SV3A * OM_L XNVmoo oxmvsnor ox xHO<na IMH HONilii aOMlMaOH are* teviNO OZ 'IWIJWifV ASIA* Ott HARTOLDMON0023474 IhluilW io-tiit Jabnviknm. Snc. *10 OHTAOI O*0 MMTMBOOK.ILUKOi> ZOb Gsor(s J. LaTiBaha.it Ph. D. Monsanto Company 00 N. Udbor|h Bird. St Loots, Missouri i)lM Dsar Dr. LaTiaakas: Bai IBT No. 441-06672 Hlatopatholo|lcal Evsloatton o( Additional Lior Socrtoas From Bata of a Two3 Yoar Chronic Oral Toaieity Sto^_^iiAroelor_W4A;_i_v^^ __ 5 .I * an sobmltttnf harosrith oor1 laboratory report datsd March 24, 1975. -np*r*d 1b coaooctioB with tbs abo to study. Vary troly pin, J. C. CoUadra Pmldtit HARTOLDMON0023475 iBIO-T3ST Ata.aMrai JU 206 RETORT TO MONSANTO COUPANT TWO . TEAK CKXCtOC ORAL TOWCITT rreDT WITH AROCLOR IMS < U.0 IN ALBINO RATS KBTOFATHOLOGICAL EVALUATION Or ADDITIONAL LIVER SECTIONS MARCH 14. 1471 DT NO. 641 06672 L latrodaction At tho n^Htt ot Dr. Ltrluku of Ha Mon Mato Company. additional iite acttoaa of Unr Iran 4 tw* r>>r chronic oral tulcltr italf et Aroclor IM to rata (IBT No. 622-07291) vara practiced lato H 6 E ataiaad aactiona and anlaaM by lifht nictaaco^r. The (eltgaria| report praiaati tba raaalti of tkla atudy. IO.TBSV 2. 207 n. Klim Mlljliriltl rt IWU|i Id ki Unt tna this w-<nhtrtw 414 apt diffar ripBcuUr Imm 4ii pwrimuly rapcrM la oar *tl|tail npnt dial Hmirdii 12, 1971. Ihwm. than ware m*w koalyo Urtr tumor* datoetad invwa aoaos W 4i nlaili at tka hl|ha*t tron&ao* Wool (100 ppm) of tka 24>k(oatk SocrlAca oktck vota oat proviso* ly raportad. Tka otkoT tmOmit-nUad laaloaa raportad ara rtfiidid > d|ari diva or kyporpUatlc la aataro aa4 thay ara attrpkoloflc moaUastatlona of an adaptlfi roapoaao o( 4a livar aatociatod witk biotmaafonxiotloa if tha taat notarial. la Roaaral, tka lattar findings vara fiaflnad primarily to taat animal* of tkaJ*. 14'- 24>Moatk SocrlAca*. and tkay vara dosa*r*latad In Ineldanc* and sorority. Hi la coodusloo, Aroclor >211 appear* ta ba allfktly tumarifanic at Laval* of 100 ppm vkaa fad caetiaaooaly la tka 41at far rva yaar*. Respectfully submitted. INDUSTRIAL BIO-TEST LABORATORIES. TNC. Boport Praparad and Reviewed by: D. E. Cardan. 0. V.M.. Ph. D. Section Hood, Patka lady ?or' V HARTOLDMON0023477 KIO-TIIT i DT No. 4U0?294 t Maaaiata *08 t ten iMUomI -- ctiaa af Um fan rate af 1ST No. 4U>0721I aai ten r*>mlsaM4 (tea for avidaaaa af aardaafoaaala- I ten nfailanf nay fiadloga far Aroclar - U&aaol*W* Iter* it rrUuw of a rhamlral aOact aa 4r llnr Uefa waa moot '1. ' orldaat at tinva of Ite H>Uaa4 oaf taradaal (H-MMk) aaertfleaa. Thla cooaiata af a hapatacaUular altaraUoa fa aft aatag focal hypertrophy which progroaaaa to anCnlir hyparplaaiau aif la a faw aAtnalt. to hepatoma* or cfaolaafiohepatatnaa. The hypertrophic call* enttla lir|a iirnati af lighttaimiaf cytopiaam which It prafaafaly rick ia glycogna aad aapoptaamie retinlao. Ia aema caaat. rtag-ahaped atroctaraa. prafaafaly ropreaoatiag whorl* of proliferative aadaplaamic reticulum. vara aaaa la Aa cytaplaam of than call*. Tfaa fayparpUatic aodalea occupied larger araaa aad oftaa eecapratiad aur rounding call* of tfaa normal liver parenchyma. Tfaay alaa caatalaad tha uma hypartrephlc caII . Thoaa lealoa* which 1 claaalfled aa faapatomaa ar chaleaglofaapatomaa vara largar aedulaa which afaowad evidence of coofloanca or aama variation in call alaa. ahapa. ataiaiag or a ductular ar adaaomatoua pattara of growth, In Aa abaanco of metaataal*. lavaaiveaaaa. aavara baaaphilia. mitoaea ar oAar evidence of anaplaaia. I choaa to call thaoa baalga tumor a rathor than malignant carcinoma*. Thara .aa ao evidence of mataataaia ar lavaatveaaa* of thaaa tumor a ia thl a atudy. HARTOLDMON0023478 'i IO.TIST 209 > M lfc Ulll With i , mi tba umli r ltatlM(|MfUl(i*nwflMc|tti, tk* Mrt Mnn kipitMtlbUr iltartttM **r M mlmal ( th* Hj ud 24-klaatfc heflAct ptrM*. UvuJtz.RuJM* Wmrt %. fticfctar. D. V. U.. U. S. Diplomat*. Amarlcaa GoUag* at Tt*riiur PathologUta ^7^ *A 0002;*/ HARTOLDMON0023479  rw* Primary thr UiImi AncUr IMS Sacrifle* bu 5 2(0 VaraoUr Ciu|i TocaI Brp>rtrt|b7 NoMir Hyp*rpU*U Hepttomi Dactolar Hyparplult Ckaluilo-Htpetaoi H*p*toc*lh>lar Nacroai* TX 0/11 0/11 0/11 0/M 0/11 0/M 0/M Til 4/10 0/10 0/10 0/10 1/10 0/10 0/10 rni 3/10 c.J,* A Coatrol S/10 2/10 Vu 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 't A - ~J . 0^ -- -4' HARTOLDMON0023480 >6 211 Prlakrr Liver Uitou Arttlw 1W Man* Sacrifice - ltti alon Vtcwlir Cku|i Fecal Hypertrophy Modular Hyperpleala Hepatoma Dectolar Hyperplaala Cholaafio -Hepatoma .-\J Hepatocellular Necroaij TI 1/A 0/6 0/6 0/6 0/6 0/6 0/6 Til 2/S 0/3 0/S 0/5 0/S 0/S 1/S Owi nn 5/9 , 5/9 V, . C/9 0/9 0/9 0/9 0/9 Control 1/10 0/10 0/10 0/10 . 0/10 0/10 1/10 HARTOLDMON0023481 7 Frlaary Llm Ltiiou Arvcter life 14 Maalfc SurlAc* . fcaW ImIw VtntUr CkU|< focal Hypertrophy Orcro T1 TO Tin \i 1/9 9/10 J/9 '> 0/9 0/10 4/9 Itodalar Hyperplasia Hepatoma Oacttbr RyytrpUiU 0/9 0/10 1/9 Vo 0/9 0/10 0/9 1/9 0/10 2/9 Chelaagio -Hepatoma Hepatocellular Necrosis 0/9 0/9 0/10 0/10 0/9 0/9 Control 1/10 0/10 0/10 O.'IO 1/10 0/10 0/10 HARTOLDMON0023482 Vo Ll*r Uilm Arwltr UK. 213 TmKal hcrUlc* Uw Vtnoltr Ckaaf* FmiI Hfpitoph|i Nodular HyparpUaU Hapttama Oactol&r HrpaxplacU ChoUaf <{) patoma Ssso TX %m ^1 Tn A,. 4/23 im *"V' 10/27 Coatrol 1/23 t/jH 3/25 Hr "iT tom *ul. iy27. % `*1 0/23 A ** am am am t*' 0/23 7/27 y-, I/.3 -Av *4 S/27 'm 0/23 t/ju km '.i- A' S/23 am << Jj. 0/23 34/27 V ' 2/27 v- S/23 0/23 >/>! IbpttsctUalar Naerecla 4/2S *3t V 2/23 ''ll *lf 4/27 *'V * /. 3/23 t\ . f\ 4f !i 000/^3^ HARTOLDMON0023483 " .A- **i , `Jjsr ' 000P.-7.*? HARTOLDMON0023484 j /i ) . * t y= .> ^ ^? . H /* rj f *i m tl n -- \ 16 * r? :: umim mil nmi : : i : i : 3^ i :(| i iUtt Vaaoolar Ckaogo Focal Hicrtili in : Focal lymyhaM laUtltntloai rml kyptrtnplif tl hapalacyt** Nofelar fcrfortrofkjr ot hapatoeytoa Focal glia pool Nvpo rjtlaal* lUpiiona 5 *8 -5ri c5* M** - fc*" r i 8! CHo la lo-hepatoma 9CZ000 HARTOLDMON0023485 I I --e \s V' i k r V ^ P cV in * a ll <-- 1 6 fc r? 2: : : : s 1 : : : JJ +^ : : : : il| 2 3 i : j 4- : i :C| : : ! * ?i?i Vaeuola r Chang* rocil Ncrili t Z Z *T * iz . Focal lymphoid Infiltration* Focal hyp*rtrophy of hopalocyl** - Nodular hyprtrophy of hopatocyto* c 3n Focil Blit Dct r HyprptaU S* * Hopaloma m9 Cholangio-hopaloma i jk HARTOLDMON0023486 000?-38 HARTOLDMON0023487 SacrWca lateral riaal (esatlBMd) /' , ;i r/.t AftOCLOft Ull TatmUtloa af ladlvMtel Unr Laalaaa la kut ] g Dom Laval fcatoo* Na. u4 Sn AH zizr 213- ft* 220" 221" 224" 233" 234" 217"*| -+230" *| 203" 204" 204" *07" that Laaloaa *8 it H* 8 h i! il c lil H Si I ks 3z 1 P - MM ' -i * u) P'i 141 *f, II * ** 1 1 AHI 24134 233" 241" 244" 270" 271" 4^4-h 2m73 T$ 243" fi+ If' 244" 312" 23 IT 274" 273*44 44-Hf. 234" 237" >02" 304" 3M" >11- 314" 240" iv*| I-* M h-1 - 4 l-l Itfj i *' H I Ml If p s L., P(P * HP It i IP-' UP<-> 13 000?99 HARTOLDMON0023488 Ot^OUO tpwa on `hhuj < 6**ia**a >i pm+rwtu 4-t-t-f ^IU r tuapaa i4V<*a r n> t lnma + DWi( nrfpug i /N i ?___________ ` <- d<.t .V ft z T 0 B 9i m 0 r i i r- ><* M <- drt c-1 v-l) d - h #i J>++ * 0A fj f! f Z J r? if *> S a ri 21 n * H ?i H t 612 ma|M^ <un *u * _ *n iapi**p"j f WJM?W |.W( i.l6' ..AM ..ll itn mv 0 * < A 1 ? A A F n f * pa* *M I**"! mq _. (paMnses) r"n.i f |aAja*n HARTOLDMON0023489 Si*iuiCucJ tlO-TIST lK MONTtSI IOAO 00*2NOl'nllOOr. UIIMOIS * Site. REPORT TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH A ROC LOR 1260 IN ALBINO R/.TS H1STOPATHOLOCICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1975 13T NO. i4l - 06672 172 V HARTOLDMON0023490 %+UUud *10 -TUT Jehtmknmi. %c Mie homtaoi *oao NOtTHltOOft.iUlNOU MM! 173 Gtaii J. Utiaaku, Pfc.D. MflBHBtD 00 N. Lindbergh Bird. St. Louis. UiMOurl 43164 . Dear Dr. Lortnahaa: ftc: IBT No. 041*06472 - Kisaopaiholofical Evaluation oi Additional Liver Sschnna Froa Bats oi a Two - Year Chianie Oral Toxicity Study at Aroclor 1240. Wa are submitting harstrllh our i ariaad laboratory rtpert datad March 24, lf75: prepared In ctBMcdot with tha abort study. Vary truly yours, J. C. Calandra Prassdant HARTOLDMON0023491 tlO-TIST c 174 REPORT TO MONSANTO COMP ANY TWO - TEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 IN ALBmO RATS KISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1975 1ST NO. *41 06672 I. Introduction At the reqtaeai Dr. LeTinakaa of tb Monaanto Compac-', additional actlona oi '.Ivor IrotB a two year chronic oral toalciry atudv ol Aroclor 1260 in rau II2T No. 622-072981 woro procoaaod into H 6 E atainod action and evaluated by li|ht mlcroacopy. Tha followtaq report preaesta iba raaulta of thia arody. HARTOLDMON0023492 IIG-TIIT d.h.^ens jw 2. 175 . Stmaorr b seat Instances, Aa epoctrust ef traafmant reltiod hietspathabgkal fladtags b tbs Uver from this ro-evaluation did not differ significantly frees Ant previously rsportsd Is ear original report dosed November 12, 1971. Hobsset. there were seres`benign liver tamers detected among seven of the <' at the eighest treatment level (100 ppm) of As 24-Uonth Sacrifice which were net previoasty reported. The other treatment-related beions reported are regarded as degenera tive or hyperplastic la nature and they are morphologic menil* stations of as adaptive response of the Liver associated with biotraasforxnatien of the test material. In general, Ae Latter findings were confined primarily to test animals of As 6-. 12- and 24-MeaA Sacrifices, and Aey were dose-related La incidents and severity. In conclusion. Aroclur 1260 appears to be slightly tumorlgsalc at levels of 100 ppm whoa fad coctlauausly la the Act for two years. Respectfully submitted. INDUSTRIAL DIO-TEST LABORATORIES. INC. Report Prepared and Reviewed by: D. X. Gordon. D. V.U..Ph. D. Section Hand, Pathology eP- * HARTOLDMON0023493 "*r A JUtadkata KIO-TIST j OT No. 422-47291 Mimihbw t km 176 J MHwl aoctlOBa of liver from imU Of OT Wo. (Zl'OTWI u< tan rt-tnlutad thorn tor evidence of ctrdaafnMli. I V kin ttMatad Bf tor Aroclor * UM M tta toflnwiag pagea: Ttan li evidence of i chemical effect n to* llnr which *11 moat evident at time of tho 12- Month and terminal (24-Month) aacrlflco*. Thli conalata of a hepatocellular alteration beginning a* focal hypertrophy which pro|roaoai to BOdular hyperplaala. amd la a fdw admail, to hopatomai or chalaafiohapatomaa. Tho hypertrophic colli contaia large amounta of light- ataialag cytoplaim which li probably rich la (lycopea aad oodoplaamic reti culum. la acme caaaa. ring-hoped itroctnree. probably repreaoadag whorli of proliferative endoptaamic reticulum, wore aees la the cyuplaam of theta eelli. The hyporpiaatic aodulai occupied larger areaa and oftaa compreiied aurrouadlng Celia of tho aarmal liver parenchyma. They alao contained the aame hypertrophic colla. Thoao toeiaaa which I daaalfled aa hepatomaa or cholaagiohepato.-r.aj were larger aodulaa which ahowed evidence of confluence or ncu variation i: cell alae. ahape. ataming or a ductular or adonomaioua pattern of growth. In the abeeace of metastaata. Invaalvaneii. never baaophilia. mitone* or ot.-.er evldenco of anaplanla. J choae to call thoao benign tumor a rather than malignant earclnomae. There waa do evidence of metaetaala o. laraaivoneat of tbeie Cuncn la thia atudy. HARTOLDMON0023494 XAAtd BIO-TIST JU. With i af tb**aoUr ad 24-Uoatfa WaniHi |Wriod. 4 ' _ . 4 ' ' 177 U- Ward R. Iliclktar. D. V. U.. U. S. Diptentt*. Ataarlcaa CoUaf* of Vatartaary Patfcalofiata HARTOLDMON0023495 Tttmnry LJror Laaiaoa Aroclor UM 1 Honk Sacrifle* - Kata Laaioo fiSBL ti TU in i VtnsUr Chu|t o/n 4/10 1/10 local Hrp*rtropfcy 0/11 0/10 2/10 Nodular H>-p*rpLa*ia 0/11 0/10 0/10 Hapatotna 0/11 0/10 0/10 Doctalar Hyporplaata 0/11 1/10 0/10 Cbolaaf 10-Hapatoma 0/11 0/10 0/10 Hapatocf llular Nocroal* 0/11 0/10 0/10 Control 2/10 0/10 O'lO O'lO 0/10 0/10 0/10 HARTOLDMON0023496 179 4 Prtmar? Lint Laaloaa - Araclor UU i Month Sacrtflca till La a ion Grata Vacuolar Chanf TI Til Tin 1/6 2/S s/s Focal Hypartrophp 0/6 0/S 3/S Nodular HyparplaaU 0/6 0/S 0/S Hapatoma 0/6 0/S 0/S Ductular Hrparplnala 0/6 0/S 0/9 Cholaaflo Hapatoma 0/6 0/5 0/9 Hapatocallolar Nacroala 0/6 1/5 0/9 Control 1/10 0M0 9/10 0/10 0/10 0/10 1/10 HARTOLDMON0023497 7 Primary Unr Uilou - Aroclor UM 12 Mcaik SacrifH# - Bat* La* loo Or--P T1 TH Tin Vacuolar Chaaga 2/4 5/10 1/9 focal Hyportrophy 0/9 0/10 4/9 Nodular Hyparplala 0/9 0/10 1/9 Ha pa tom* 0/9 0/10 9 '9 Ductular Hyparplaaia 1/9 0/10 2/9 Cholaaglo-Hepatoma 0/9 0/10 0/9 Hapatocallular Necroala 0/9 0/10 0/9 ISO Control 1/10 9/10 0/10 0 '10 1/1C o/.o O'lO HARTOLDMON0023498 ef .if % . I*J Mwrr Uni Uiliii - fcwtlwsiUW-' ' Tmlal Socriflco - Xmti % . - Grtmc TT TO TD Control VicMitr Chaafo S/25 4/23 ie/27 1/23 !"eii Hyportrvpbr J/2S 10/23 1227 0 ;23 Nodilu Bparpluli HapMOBtB Duttlw HyporpUoU 0/25 0/25 4/25 9/23 0/23 5/23 7'27 9/27 14 '27 1/23 0/23 5/23 Cteluflt-Htpaiuu 0/25 0/23 2/27 0'23 Hapotocollulnr Nacroau 4/25 2/23 4/27 1/23 I 000251 -- r> f HARTOLDMON0023500 _ */ s o <5 n ii if5 'yA:'KOh <4^*a4 *u41 w:ft .<S3,S2,aSS-j uS= *8=SnK i: ---------r y3 ?zJi Vacoolar Chaafa Focal Nacroala f4 Focal lymphoid Infill raliono Focal hyporlroptiy of hapalocylaa Nodular hypertrophy of hapalocylaa Focal Blla Dcl llyparplaala St It |8 L h C| si i llapaloma Cholang lo-hapalama Cj HARTOLDMON0023501 1 it': o o o u\ CJ HARTOLDMON0023502 f> r? ^j ssshre^s sssstssstjjssssgsssresssre 3 1 *i J I 5 1 i j[ 5 2 2 3 3 3 S3 3 3 3 3 3 3 ^33 333 3 : i ; J fEJ Vimbr Focal Mrcrucla Focal tymfloU Ikflltratlaaa ofrocal kyyoMrofrtiy hopalocytoc Modular fcyf Hcal>T of hcpalocftoc Focal |ll Duct llyyorplacld ' ') ' lUpoloma ). t l s 3 It |8 Is a ChoUn|lo`hepatoma Cn c fS 2 0 G 0 HARTOLDMON0023503  S- ABOCLOB UW 1 akntlaUoa a* laAH44wa I Unr La lota taJLata Sacrifice lotonral Doao LotoI Aalma No. u4 Sob Uw UitoM m 3 ci li*! n Si ri" e II a 1 1 r 1u Flaal (cnnrl--i it) o AS xizr X13" Z20" 221" 224" 233" 234" 219" ZJ0M 203" 204" Z04" 207" Ad 24134 ZS3" 241" 244" 270" 271" I 272" I Z751' : 243" j 244" I 312" 23 ir | Z74" I 273" I 244" I 297" 302" | 304" | 301" ' 311" j 314" ' 240" ** P P P P P P ,.*7 . *-( I lit. ijWffB ~ *3 ; *&,' .. 1 ' S&~* -.i,r'vj&r* ^ 000255 HARTOLDMON0023504 5 ol o *0 w o i l if I 5'sra * * *z { [I Tiff VintUr Ckaoye facilHicrMlt Fil lymphoid laflltratlaoa rocal hyporltopliy of kapilstyMI Nodular hypertrophy of hapalocyiea Foul Blla Duel Hyperplaaia llepaloma Cliolanyio -hepatoma HARTOLDMON0023505 Am4uUual tio-TlfT Jab&aAmu. She... < rtONfUU low MO*TrOQ.IILINO'I mm> 077 REPORT TO MONSANTO COMPANY TWO YrAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR ISM IN ALBINO RATS KSTOPATHOLOCICAL EVALUATION or ADDITIONAL UVSR SECTIONS MARCH 24. m$ IBT NO. 441 . 0t>o7: / ,.. . . .J hVt' IO-TST .ftrfeuf'niK. Jiao. MM raomraaa MM NOITMt*OM. ikUNOS HIM July 1. 1075 078 Goorya J. Laytaafcaa. Fb.D. 100 N. I tndhf|h Bhrd. St. Loola. Mlaaouri U144 Ra: 1ST No. Ml-04472 Klrtopalhnloglcal Iraluattoo a< Additional Llvar Soctiana From Rota a< a Two - Yaor Ctoonle Oral Taatdty Study erf Awthr 1240. Wa ara aohoritttoR harawilh oar rarvlaod laboratory raport dMad March 24. 1?75, pcaparad is connocdan with tha abort study. Vary truly years, --i. J. C. Calandra JCC/fd HARTOLDMON0023507 ,W-lW BIO-TtST 7f REPORT TO MONSANTO COMPANY TWO YEAR CHRONIC ORAL TOXXITY STUDY WITH AROCLOR 1260 IN ALBINO RATS HtSTOPATHOLOGICAL EVALUATION OF ADDITIONAL UVER SECTIONS MARCH 24. 17} IBT NO. Ml . 06472 L Introduction At the roquast of Dr. Lavinakas of tfaa Monsanto Company, additional tactions of '.Ivor from a two yaar chronic oral toxicity study of Aroclor 1260 in Tata (IBT No. 622-07246) wore procoaaod into H k E ataiaad aoctiona and avaluatad by li|ht microscopy. Tho following raport praianta the rasuits of this study. jd, I S*!! S hjtfcManxlAil 000253 + NMMMKtfC H. HARTOLDMON0023508 III ^11 f tl0TtSY rfb.. .huSti &f.ki~w"I trCr, l" e la 2 080 from that prarrtaunty i^aat ta aw aaiglaal nput Afad Waytiir U. 1T71. Thm wn ura hapaaaoma htah4 wa| aoraa at tha ataili u h highest triifna Went (1M ppmj at Ae It Heath Sacrifice. No hepaw cellular r irrimiaa wn aba aired. Tha athar Ira atmant-related laaiooa reported ara regarded aa degaaeratlTe or hyparplaatta la natwa aad they ara morphologic anlfaatattaaa at aa adapt!aa raapeaee at tha brer aaanrtatait arith btotranafarmatloa at Aa wet aaadal. la general, Aa lattrr dotla|t were nnnfinad primarily to Wat tnl~r*T at tha b*. 12" aad 24-Uoath Sacrlficee, and they were doaa- relatad ta larirtaara aad arrarity. la coaclaalaa. AracVor 1144 daaa not appear w be caretaopaalc u> raw fad far two yoara at levala W aad taclttdtag 104 ppm. Raepectfully tabat Had, INDUSTRIAL BIO-T1ST LAB04LATORUS. INC. Raport Prepared aad Raaiamad byt O.c. Gordoa. O.v. VL . Ph. 0. Sactloa Head, Pathology i HARTOLDMON0023509 HARTOLDMON0023510 U'aJr.&cMI' Vtrtf H. IldMi. a v. U., &L s. DtltMHM. Amrteu CtlUft o( VMtrtitry PiriMtofiaw . . . i- 000262 * V.* * ''^f14 .. -v HARTOLDMON0023511 A' . ' ' ' V ' t: * / ir'v j:. r Uaat Laataaa Aaaalar UM V`.?J - 1]Maa* ttriflca - Kata *' ' Vwwlu Ou|< rocal Hypamaphr KaMtr HrptrpUlU Hapnotna DvaIu HyparpUata CfcaUafia-Haparoma Kapatocaltalar Macraaia Vj TX /II 9/11 0/11 0/11 0/11 0/11 0/11 m 4/10 0/10 0/10 0/10 i/io o/to 0/10 thi 1/10 1/10 0/10 0/10 0/10 0/10 0/10 Caatral 2/10 0/10 0/10 0/10 0/10 0/10 0/10 i HARTOLDMON0023512 ^;w * v',. -;V '^"V >* '. ' y .<- ', . - ri*l -*- . y. 4 ; ii4 0 Vrtemrf U** Trt-- Araclar UM Liriw 4 Um6 SmtUU* - Iw ' T1 Til fin tux VicmIii dui|* 1/4 2/} s/s rosi Kry*nfhr Notmli* Hy^r*U*L* 0/4 0/S 3/S 0/4 0/S 0/S Ovetdir KrfiipUili 0/4 0/S 0/S 0/4 0/S 0/s dfUagta-Haytom* 0/4 0/S 0/S Hl*Mtllibr Ntcmlj 0/4 1/S 0/S CoMrol 1/10 0/10 0/10 0/10 . 0/10 0/10 1/10 HARTOLDMON0023513 CHb \. *rimary lire* Lsslaas - Areclor UM i Meath Sacrifle* - Rats L*lcm Groce TI T3 Tm Ccctrol Vacoolar Chase* i/9 5/10 5/9 1/10 Focal Hypertrophy 9/9 0/10 4/9 0-1? Nodular Hyp*rpla*l_ Hapatooaa 0/9 0 '10 l, 0/9 0/10 0 '9 3 10 o :o Doctalar Hyperplasia Cholaaglo -Hepatoma HrpatoceUalar Necrosis 1/9 0/9 0/9 0/10 0/19 0/10 i/9 0 '9 0/9 ! 10 0 10 o :o HARTOLDMON0023514 I 096 . Prtatn Ufit UaMu AraclM UM Ltoc Taraiaal Merlfioa Kata V Ortwm T1 TU Tm VacKltr Cku|i Focai Hypru tns iUS 4/25 14/25 10/27 11/27 No4iUi HptrpluU o/2S 5/25 7'27 IbfllOBtt 0/25 0/25 5/27 Doctular HyprpU*ik 4/2* 5/25 14'27 Choln<io-Hptott* 0/25 0/25 2/27 Ki^tMdhUir Ncro-. 4/25 2/25 4 '27 Coalrol 1/25 0/25 1/25 0/25 512 5 0 '2 j . t '22 HARTOLDMON0023515 IS 75 ^ MNM II.J yM&IlijJiMSIili i ; i ! i s x t 11 t * 11 ??* VttMUr Cku|t ractlNiciHti Focal lymphoid lellliralleoa Focal hypertrophy ol hapalocylaa Nodal* r hype rtf I hapalacytaa Focal Hilo puci Hyporpiaala IWpalmna Cholaapla-hapalom* m u l *|an ia net nK |n*opo>r/lFp l f oonw |oTi I 80 HARTOLDMON0023516 j* fa rt i -- 3 2 2 2 5 -f ^ j 1 i jS : : t^ t M> ^ n 21 -- & ]? r trSCyw : i|; : : *| i j g ri; VamUi Ciu|a Focal HtcraiU Karel lympluM latlUrailaoa Focal hyp* rlraphy of hapa'ac/iaa Nodule r kypai/rtfUf of kayilotylal Focal Blla DaeI HrftrUaU^, llvpaionu. i ------- latf. Cholaofko- Ft fl r Ii -! ,> , .4 .- 4\ f / $.< C? HARTOLDMON0023517 00026P - * 000269 HARTOLDMON0023518 tcafl HARTOLDMON0023519 AftOCLCft 12*6 TtbuUtiM ot MWidl Umt Loilwu la Rata 091 Sacri/ic# Intarval I Doa# J*olma i Laval No. aad Saw ? 1y riaoi ; as (coimanad) ' 212T 213" 220" 221" 224" 233" 234" 219" 230" 203" 204" 20a" 207" Uvar Loaloaa >6 J i0 2 1 f: i i! c i I 8 I:- i !i 3s - *3 * It u XI iL 3* *uw03. 9U 9 L*____ 1 2 5 i * 9 ?*3 W i | 1 i I ! 1 .1i Am 24124 233" 261" 1264" ! 270" i 271" ! 272" 1273" 243" ; 244" 312" 23 LT | 274" ! 273" ! '04" I 237" !ii 3J0024" | 300" ' 311" I 314" ! 240" III I i p p p p p p 13 HARTOLDMON0023520 i<S Sp *-* -aA* JAN 3 A D -a HXHTXW O - * M * Oj :*1-: U> 3: ys m a. 2 Vinelir Chango I'MilNucrotti 9 *0 Koctl lyrn|.hulJ lalllitailoua Focal hyp*rtrophy of h[ulucyl i Nodular kyporlrvplty at hapaiocylaa Focal Bila pucl Myparplaata Ifopalonia ifc sR! 8-5 c9 n r 0 C Itole agio-bapalonia N I I HARTOLDMON0023521 Uiion Vacuolar Change Focal Hypertrophy Nodular Hyperplaaia KUptttomi OictuJar Hyperplaaia C ho lan gio-Hepatoma Hepatocellular Necroaia Primary Linr Ltttou - Arodor 1240 TiratUil ecriflce * Hate M 1f5 1/25 2/25 5/25 Oroay m Till 121 121 1/21 2/20 4/23 8/27 7/24 10/27 0/25 0/25 1/25 1/21 4/20 10/23 9/27 6/24 11/27 0/25 0/25 0/23 4/20 9/21 2/27 3/24 7/24 0/25 0/25 0/20 0/21 0/27 4/24 5/27 1/25 1/25 4/25 2/20 5/21 1/27 3/24 14/27 0/25 0/25 0/20 0/21 0/27 0/24 2/27 1/25 4/25 0/23 0/20 2/21 2/27 11/24 4/27 093 Control 12 1/21 0/24 0/21 0/24 1/21 0/24 0/23 ill* 5/23 0/24 0/21 0/24 1/21 1 - Firat twliutini of original elidea 2 - Re-evaluation of original >1Mm 3 - Evaluation of additional ttaeue aaettona HARTOLDMON0023522 MAkf IO-TIST VrlMiT Uoar taaitmm - AtOCUM - 1240 Taraiaal SacrlAca - ftato a-anlmtlai af 0*VmI SJdM Vacuolar Quaga Focal riypara ophy A-I 2/25 /2S A-2 2/20 4/20 A-2 7/24 4/24 Nodular Hyparplaota BUa Duct Kyparplaala Q>olaogto-hpc>naia Hapatoaa 0/25 1/25 J/23 0/25 4/20 2/20 0/20 0/20 2/24 2/24 0/24 4/24 Curol 0/24 0/24 0/24 4/24 0/24 0/24 HARTOLDMON0023523 HARTOLDMON0023524 UM tlp.TIST 1 6 HARTOLDMON0023525 O iJO f cr-. j ! 1 F l r HARTOLDMON0023526 E X H I B I T HART OLDMON0023527 ( OrAuUaL tio-Tisr fnUvttmm. She. i*t# rioMuai *e*e NOITHIHOOI. lkUMO tMt] 239 REPORT TO MONSANTO COMPANY TWO - TEAS CHRONIC ORAL TOZXCITT STUDY WITH AROCLOR IN ALBINO RATS KBTOPATHOLOGICAL EVALUATION OF.ADDITIONAL LIVER SECTIONS MARCH 24, 197S I3T NO. 441 - 04*72 HARTOLDMON0023528 210 t SmiatBiit tIO TUT ftrifirtTfrif She. 1*1* PaONTtOI *OAO NOirxiieo.iiuMii tNti hlnrcb 24. im Gsorf* J. Ltriuku, Pb. D. MooMBta Company BOO N. Lladborfh Bird. St. Losli, Missouri 631S& . Osar Or. Lsvinakaa: ' Re: IBT No. fr4l>06672 - Hlstcpacholofteal evaluation oi Additional Liver Soctlooa From Rata at a Two * Tsar Chronic Oral Tsatc.tr Study ol Aroclor-W4. 17.HI -1TT aro submitting herewith oar laboratory roport dated March 1975: prsparsd In conaoction with tba above study. Vary trulv yoara. J. C. Calaedra President HARTOLDMON0023529 BIO-T 1ST i -W *41 REPORT TO MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITT 3TVDT WITH AROCLOR 1264 ' IN ALBINO RATS histopathological evalcaticn or AOOITIONAL OVER SECTIONS MARCH 24. 197J 1ST NO. 641 06672 I. Introduction At tha raqua at of Or. Ltnnikai of tfci Monauto Company, additional aacttona of Uvar front a two yaar chronic oral tonicity study of A roc lor K66 '.ly T :n rata lIBT No. 621 -07276) wore procaaaad Into H 6 E atainod aactiona and avaluatad by light mieroaeopy. Tha following raport praaanta tha raaulta of ihia atudy. HARTOLDMON0023530 lO'TIST 2 242 --( In mot truaanl-Nl>M2 Mrtcyi>ih|lnl fladii|i la the liver from tide re-evalnntioB Aid aet differ aignlfUantly from that previously reported la oar ori|iMl report dated November 12, 1971. Hiieint, there were three lenip Ueer tamore detected amopg three of the ulsali at the highest treatment level (100 ppm) of the 24-Month Sacrifice which were aet previously reported. The other treaeaent-reletad leeioaa reported are regarded aa de generative or hyperplastic la aatore and they are morphologic manifestation* of aa adaptive reapoaae of the liver aaeociated with biotraaaformatioa ai the teat material. In general, the latter (lading* were confined primarily to teat animal* of the final eacrlflce and they were doee-related la Incidence and aeventy. In conclualoa, A roc lor appoars to ho (tightly tumorigeaic at level* of 100 ppen when fed contlaouely in the diet for two year*. Respectfully auhmltted, INDUSTRIAL BIO-TSST LABORATORIES. INC. Report Prepared end Reviewed by: O.C. Cordon. O.T.M. .Ph.3. Section Hoad. Pathology HARTOLDMON0023531 IO-TIST t IBT No. HI-07Z*I Uoaiuto J l kit* uunlitd idditlAMl McdMi of Utn tM nil of Staff Number 022-07240 ud hare t*-*nhut*d tbam (or eeidaace of ; inc tabulatad my flidh|i far Aroclor f&t M tha following pages. Thors la oridooeo of a chemical affect aa tha liver which l* moat pronounced at ttma of tha terminal (24-Month) sacrifice. This conaiita o: a hepatocallular alteration beginning aa focal hypertrophy which progrv to nodular hyperplasia. and la a few animals to hepatoma or cholani'.ocepatocna. Tha hypertrophic eella coataln lerfe volumes of Ught staining cytoplaem whteh i* probably rich la glycogan and eadoplaemic reticuhan. la soma sections, .-.c,-- haped truetore e, probably representing wbarls of proliferative endoplasm..: reticulum, ware taaa la tha cytoplasm of thaaa cello. The hyperplactic nodulee occupied larger areas aad often compressed eurrounding cells of the normal Urer parenchyma. Theee nodules also contained the same hypertrophic cell*. Thooo losioas which 1 claesUled as hepatomas or cholaagiobepatomas larger aoduloa which showed oridoace of coafluence or lomt variation :n cel! i.i;. (hops, staining or a ductular or adenomatous patters of growth, hi the absence mataetaels, laraalvoaoae, severe baeophiUa, ttutoeee. or ether evidence of ana plasia. 1 ehosa to call thaoa beaiga tumors rather thaa mallgnana tumors cere.n- onail. Thors erou so evidence of metastasis or larailveeeae of these tumors in this study. HARTOLDMON0023532 IIO-TIST 244 " Tl 111mli In pilalil mil tint 1HIIF .......................10a nil *m hujuim^ With axcoptloa of tho vacoolor <kti|i la tho cytoplasm of Wpatocytoa, tho mort severe hepatocellular aUeretloa* wore coaflaed to oaimale of tho 24Uootfa Sacrifice. Word R. Richter. S. Dipl., Am. Collage Vot. Path. i HARTOLDMON0023533 Primary Unr Lnou - Aroctor 126* 1 Month Sacrifice Rati UliOB Croo TI Tn TOT ' .easier Chany* 0/8 2/10 1/9 Focal Hypertrophy N'ouolar Hyperplaaia Hepatoma 0/8 0/8 0/8 0/10 0/10 0/10 0/9 0/9 ` 0 '9 Ductuiar Hyperplaata Jholanyio- Hepatoma 0/8 0 '8 0/10 0/10 1/9 0/9 Hepatocellular Neeroeia 1/3 0/10 0/9 Control i. 10 c 'i: o i; ,9 * e : :u :n 000284 HARTOLDMON0023534 r * K Prirr-ery Lirer Lelon* - Aroclor 1284 ft 8 Month Secriflco - Ret* L*ion TI Vacuo Ur Chaii|* Tocel Hypertrophy Noduler Hyporpleole 0/10 0/10 0/10 Hopetome Guetuler Hyporpleiie Cho len| io-Hopetome 0/10 0/10 0/10 Hopetoccllular Nocroti* 1/10 TQ 0/1 0/8 0/8 0/8 0/8 0/8 0/8 Grom TUI 0/9 0/9 0/9 0/9 0/9 0/9 o. 246 Control 1/10 0/10 0/10 . e/io 0/10 0/10 l /10 *% a G30285 HARTOLDMON0023535 7 Ptiatir Uf*> Utlsu Aroclor 12M 14 Month Sacrifice - Kota Laaloa TI V a coolor Change 2/10 Focal Hypertrophy Modular Hyperplaaia 0/10 0/10 Hepatoma - 0/10 Ductalar Hyperplaaia 0/10 ChoUnfic-Hapatoma 0/10 Hepatocellular Nocroaia 0/10 TU 4/10 0/10 0/10 0/10 1/10 0/10 0/10 Group tm 0/7 1/9 *1 0/7 0/7 .0/7 0/7 0/10 Coaml 1/10 0/10 0/10 0/10 1/10 0/10 0'10 pv r 0. wt <<*- HARTOLDMON0023536 Primary Unr Liilou - Areclor 12M Tormiaal Sacrillc* - Rata g * 248 Lootoa Vacoolar Chao** Focal Hypertrophy Nodular Hyporplaala Hepatoma Ductular Hyporplaala C ho laa*to - H* patoma Hepatocellular Necroaia 3* ft **Vm UTa, 3 1* Grog; TU *> 1/30 3 Tin 3/20 / *? 1 Control 1/23 2/11 %i#/n 1/30 S/20 *A-o ^u 0/21 om A*,*TM 2/30 8/23 "'f** ' t/ 1/21 0/11 */> t "%7 0/30 2/:a A/ Ml /*<> U a '2i s/si^/Tai 1/10 1/20 >ho *, S/21 0/11 1/31 @> 0/30 h* v'< Yiriv, 1/10 G> G> 6> 1/20 0/20 <3 /> r> 3 3 23 l 21 X 0V, .H I4* ml -- ll I a -y, 4. v ,.~L .14 * /* *. <u.r 000287 * HARTOLDMON0023537 fr;, ** p) . Vs. ,-1 I $:> 3n x og 4 a *4 ss s f w w m w i ^* : : ; : "I : : :O ^ *4 gO 4# H 9> j e. : i ii 4-45) 4*)* X 3 *. A o o o N CD CZ> Tri oO bAb o o 040^01I M*4 *M4 *M4 *-4 g ^-4>-^4>-4>->I 41 Ol 4 O' ib4U(4W*A4<4 Q4-4^4g Ji S'? 3: nfi ^ion !^lftttni*me ivg*ctumo1i.*' Focal Mcroiit of hapatocytea Focal lymphoid Inflltralluna & Focal hypertrophy ol hopalocyloa Nodular hyparplaala of hopatocytoa Focal bilo duct byporplaala !i E1 g7 * hr *i. Hepatoma C.ho la oglu-hepatoma toticulum Cell Sarcoma! metastatic Mammary tumor, molMiitilc ] TtfUnu( l.mu. N (O HARTOLDMONOQ23538 AROCLUR . Tabulation ai ladtvidaal Unr Leaiana ia (at* Sacrifice 0N Animal) Interval Laval No. and 1Sax 3 i0 3S J1 H ~X u_ ui & Month | C-I iom* ! `pp* 1011"1 1011". 10U"1 .1 C> tS>>--- . 10 IS" 1 1026F, 1027" , 1024'' j 1029" ! 1030"J 3s 11 nV l k9. Uxu_l*ilaai I 4-*1 r & hi M i II f i u59. " 10 z ma c-n 1041M! 1 lOppm 1042" 1041" ` 1045" j 10S6F ! 1057" 1058" . 1059" | I c-az 1071M. lOOopm 1072" 1 1073 ' ' 1075" I 1 I l.X v 10S6F ' 1017" ' 1088" 1089" 1090" Control 501M . a 802" | <> 803" 1 804" i 80;" 816F 917" i 318" 819" 820" 0 1 i o ? X 10 250 sJ -l-!` 5 i 3 j' 3 !| " Ef f 000.?89 HARTOLDMON0023539 AROCLGR - lit*' of laKividaal Uni Laaloaa la Kata 11 T o U n g e cU iU , (oc| * -. - 4 000230 HARTOLDMON0023540 I AROCLOR . 1260; TthiltHw at MlvUaal Uftr Lsaloaa la Rata 252 12 Sacrtiica Data (Aalmal lnt ml Laval No. aad 5as St J a II R Li aJ J; 3 u u J'S ^o II ii__ ; i- x -a 24 Month jc-I sum! i ,PP*n 366" | S9t'H "/' i 567*' 5*9" 1r) + K' 572" 576" 580" 588" 889" 395" MIT f 604" 603" S'JBZ" l*l-Or.ml (atl ] 1*6 (.ajr V8J" 607" fo* 'v *i>t I 608" 609" ^naiw! " tk a *.*4 u-v- tai nr l.n4tfljp 613" |rt* 621" 627" 628" 6;i" 6:*4M 635" ^606"^ UlO"" f*20,,J y626" T6J"1 Lira r Laatcaa IS 1! H if i* 0 tgk L R Z K> 1*>* H H* <' I**) 2 1 2 *R j j* 31 si 5 si H uac Hpim ! 1^*1* I4 1-rr* lu. A-J., -t Lttij t J 000.^91 HARTOLDMON0023541 TiT- -- MN l* * * J to it l fs ----------------4U jl----------------- jl---------f--1~_ 044iHHOoodjiiiiii^uiWiIiI,Kfruiui rr ,`i.``1 V* * *.~ ~. :" ~" ` i-~.f TFJ t ' ^ : ; * : fi.~:.~: ii.:'.:'^: ':^: |CytopUitnic vAcwoUtioa of iMpttocyUi Focal aacroala of hopalocylaa T" T" Focal lymphoid laflllrallooa Focal hypertrophy of liopotocyloa h H "TT t j_ Nodular hyporplaaia of hapatocytaa Focal Mlo duct hyporplaaia 'ape tome E 1` *Z Va i w Chol*n|lo-hapatoma Raliculum Cell Sarcoma] mataalatie Mamniiry tumor, iiivlualatic I - I i it* I i * i*i , I* >i I u< Co HARTOLDMON0023542 [ A HOC LOS TabaWttaa sf Mlfltal Unt UiUu la Bats {$4 HARTOLDMON0023543 S a crifice .) ___ 1 s* ,Y (V f t rf l 3 r* pT 8B rv 8*i* sE?| n a s i * * -S 1 Cylc^Uwnlc Vkcuolaliaa ot hafkiocirlti F*mI Meroili of Iwpalscftu H ? a. i Focal lymphoid laflltratirwa x Focal hypotrophy of hapalocylaa E2 Nodular hyparplaaia *0 ol hapalocylaa J Focal hlla duel hypa rplaala r o 0 llapalaina V m S Cholanpio-Sapaloma i Mottculum Call Sarcoma molaaUtle Mammary tumor, mUuilc T IrtiiKei'U^iit, fui.it <N/> b< Ul f HARTOLDMON0023544 JIft&aUtal tl O TIST JabMaloMeS. %& 110 MONTMI lOAO . NO*THMOO.IUINOl* tOMI 155 REPOST TO MONSANTO COMPANT TWO TEAR CHRONIC ORAL TOXICITY STUDY WITH A ROC LOR 1242 IN ALSENO RATS H3TOPATHOLOGICAL EVALUATION OF ADDmONAL LIVES SECTIONS MARCH i4. 197S I BT NO. 641 - 0667J / )VW* ^.L^gr.y v y > K-. . . - - .* ""'.L __ \S.'~ 00029;' HARTOLDMON0023545 9*kut**i 10-TIIT Jsie*t6*U. %c 1IM MpNTAOl tOAO ' NOrHtoo. iiunoi* MMt 156 Gooryo J. Lorlaskas. Ph.D. Uonsanlo Coapany tOO N. Uadboryh bird. St. Louis. Missouri 43164 Osor Dr. Loriaskas: Rsi DT Mo. Ml-06472 - Histopatholoyical Evaluation el Additions! Livor Sections Froa Slats of s Two - Year Chronic Oral Toxicity Study d Aroclor 1262. Wo sro subaittiny horowtth our rovissd laberstory ropert daud March 24. 1775; prepared la connection with tbs aboro study. Vary truly yours. J. C. Calandrs Prosidsnt HARTOLDMON0023546 BIO-TIST ' JU * 157 REPORT MONSANTO COM. ' TWO - YEAR CHRONIC ORAL TOXICITTY STUDY WITH A ROCLOR 042 D? ALBINO RJtTS HISTOPATHOLCOICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 197! :BT NO. 641 - 06672 L Introduction At tfaa raona it ai Dr. Laa-.nikaa ol tba Monianto Coir,pa try, additional itcr.om of lirtr from a two - mr chronic oral tonicity atodp al A roc lor 1242 tn rati lIBT No. 622-072981 wora proconad late Hi I mtaod lactica aad avaluatad hr '.ight trucroicop-,-. Tha foi'.owiaj raport praaaats tha raiults of thti itudv. HARTOLDMON0023547 JUiW tlO-TIST 158 IL Suamar^ In not uutucM, Ik* spectrum of tr a* ft-related lil*b|i*l1>aliiglc*l ta th* liver from this r*-evaluation did not differ significantly from that praviouslv raportad In onr original raport datad Novmb*r 12, 1971. However. thorn war* ehraa benign liver tumor* detected thraa of tba animal* at th* highaat traatmant level (100 ppm) of th* 2 4-Month Saerlfle* which war* aot previously raportad, Th* othar treatmaat-ralatod lesions raportad ar* regarded ** de generative or hyperplastic in nature and they ar* morphologic manifestations of aa adapdr* raapoaa* of th* liver a*ociat*d v-.th biotraasformatioa of da* to*: matariaL la gaaarml, tha lattar finding* war* <-oaf:aod primarily to t**t amma.s of tha final lacrilic* and they w*ra doa-ralatad La Lac Id* oca and Mnrr. In conclusion, Aroclor 1242 appaan to ba (lightly tumor!goaic at Lovol* of 100 ppm whoa fad coatiaoualy in th* diat for two roars. Respectfully submitted, INDUSTRIAL. BIC-TSST 1-ABCRA70RLE5, INC. Report Prepared acd Rsnewad by. O. E. Cordoa. 3. V.M. . Ph.3. Section Hoad. Pathology Report Approved by; Manager. Tjmcolog'J if HARTOLDMON0023548 < 1 Wmlkml 110-TIST IBT No. 622*07296 Monsanto im. } ' f_ 159 t have examined additional Mctlou of liver from rat* ti 9tdy Number 622*07298 and have re-evaluated them for evidence oi carclaayaaaals. : have tabulated my findings (or A roc lor 1242 oa the following pay**. There it srideace of a cbeanlcal effect oa die liver which la moat pronounced at time of the terminal *24-Month) eacriflee. Tbia coaaiata of a hepatocellular alteration beginning aa focal hypertrophy which progressea to nodular hyperplasia, and la a few aaimala to hepatoma or cholsngiohopatosa. The hypertrophic cells contain large volumes of light staining cytoplasm which is probably rich la glycogea sad endoplasmic reticulum. la some lecnoet. ring shaped structures, probably representing whorls of proliferative endoplasmic reticulum, ware seea m the cytoplasm of these cells. The hyperplastic eodulss occupied larger areas sad oftsn compressed surrounding cells of the aormal liver parenchyma. Thee* oodutas also contained the same hypertrophic cells. Those leatons which l classified aa hepatomas or cholaagtobepatsmae were larger nodules which showed evidence of confluence or somo variation m cell stso, shape, staining or a due tula r or sdsaomatoua pane n of growth. In the absence of metastasis, invaaiveneas, severe basophilia, mitoses, or other evidence of ana* plasta. I chose to cal! these benign rumors rather than .malignant turners carcin omas , There was .to eviieoce of metastasis or tavaslvenass of thssa rasters :.-.ij arady. J* m L HARTOLDMON0023549 JU*td IIO'TIST i . JU 4 160 With wjpIWi at tha vacular cfcaaffaa In tha cytoplaa .at hapaaacytta. tha or* aavara hapahirallnlar aharattaaa wara caaftead a aataala at tha 24-Uaath Sacxifica. (Jt*dB Ward R. Rtehtar. D.V. U..U.S. DtpL. Am. College Vet. Pith. 9 yTjwr^- , -*' - >A.V. - -a * OQQrr- HARTOLDMON0023550 Prlnirr Llrar Laaiooa Aroclor 1242 S Ummtk Sacrifle* kata- La atoa Croo TI Tn Tm Vacuolar Chaaga 0/S 2/10 1/S Focal Hypartrapbf 0/8 0/10 0/S Nodular Hyparplaala 0/S 0/10 0/S Hapatoma 0/S 0/10 0/s Doctvlar Hrparplaaia 0/8 0/10 1/S Cholaaylo> Hapatoma 0/8 0/10 0/S Hapatocallolar Nacroala 1/8 0/10 0/9 IS1 Control 2/10 0/10 0/10 0 10 C-'10 O-'IO O'lO \ .; 0^0301 # HARTOLDMON0023551 Primary Unr Lesions Aroe lor 1242 4 Month Sacrifice - Rats . La Ion Group t: tu Tin Vacuolar Change Total Hypertrophy 0/10 0/10 0/1 0/8 0/8 0/9 Nodular HyparpUsia 0/10 Hepatoma 0/10 Due tular Hyparplaeia 0/10 Cholaag io-He patotna 0/10 Hepatocellular Necrosis I MO 0/8 0/8 0/8 0/8 0/8 0/9 0/9 0'9 Z *? 0/Q t Control 1/10 0. 10 ' 0/10 - 0 10 0. 10 0M0 I 10 00030:' 4 HARTOLDMON0023552 Primary Llvor Utiou - Aroclor 1242 12 Month Sacrtilca - Itti Lotion Croon TI Tn t m Vacuolar Chang* 2/10 4/10 0/4 Focal Hypertrophy 0/10 0/10 1/9 Nodular Hyporplaala Hapatoma Ouctular Kyporplaiia Cholanglo - Hepatoma 0/10 0/10 0/10 0/10 0/10 0/10 l /10 0/10 0/4 0/9 0/4 0/9 Hopatoco llular Nocroaia e/io 0/10 0/13 7 163 Control 1/10 0/10 0/10 0.10 1/10 0/10 3'13 000.^03 HARTOLDMON0023553 Primary Hrer La done Aroe lor 1242 Terminal Sacrifice Rate J64 Legion Vacuolar Change Toe el Hypertrophy Nodular Hyperplaeia Hepa:oma Due ruler Hyperplaeia Cholanglo-Hepatoma Hepatocellular Nee self TI 7/11 2/11 0/11 0/11 1/11 0/11 1/11 Greg TU Tin 3/10 9/:o 1/10 /:o 2/10 3/20 0/10 2/:a 1/10 0/13 i/:o ' i/:i 1 10 o,;o Control ! :3 i :j l :i i :: 5 :i o :i : .. HARTOLDMON0023554 760" 796F 797" 790 00" Con trol J6M 37" ja - 39 40" ! 76F 1 77- U. 79" | JO1 I HARTOLDMON0023555 0 l nO P O rn 0n f2 * *o5 ST? 3: w*-'*----ooooo OOOOOOOOO 5 5 5 -* *f ; - ; oooooooo yiUUIUI^^^^ : J si : : (' oo o< o* N N ti N I O: : >: *:1 *^t I Oooo 7 b* N M Jpg *& u 6. 0 J s : UL Cytpplaainlc vacuolaI** 0( l>oMtocvtaa Focal oocroola of hopatacytas + Focal lymphoid loll Itratlona Focal tiyporlrealty of hapatocyias r Hm f V i * nIs MJ Nodular hyparplaala of fcapalocytoa >p * r* Focal bfW duel byptrpItaU ii llapa|oma Cliulanglo-hopaloma Iwllculum Call Sarconu malaalallc Mammary tamer, motaatallc ... . lulaunaclaala. focal 1 to. HART OLDMON0023556 AROCLOR . 1242 Tabulation of tadhrl^aal Unr Lailoaa u Rat* SaerUlc# I Dot* AMmall Intarral 1 Larol No. 1| "So<n1 ihm 1 *Ui i' -s! Oj J :t ' ' ' ("_} tIt** 12 Month! C-I : lppm : 71M j 72" * , 73" 74-' 11 75'- i 76F 77" 78" ! 79" ! 80" ! C-n 81M * ! 1 lOppm 92" . 83" 84" * ; 89" ! 86F . 87" 88- 1 89" 90" * ' C-n 91M *9 i! f>S;! ($h * 0 o ~2 is fms9 * r 2m 2 * i n M i\ i f* i t 2 X t I 1 !" z (J9* -c l 1 : r. 1^ 1! 1| 1: i: i .; - 1 11 i; ( ! i 93" 94" 95" : 97" 98" 99" Control . IM 2" 3" 4" 5" 6r 7" 9" 9" , *' * 1 *1 i ! i H7 9 c0 1 2 i, a* 7 3 uaMoo. 1 1 Se ia ' '- I IIM U lU lIt 11 HARTOLDMON0023557 ABOCLOA . 1342 Tabulation of ladlridual Llrar La*leas in Kata 168 >2 HARTOLDMON0023558 ............................... i 1' ! *\J\*3* S *>d 0 1 ".............................................................................................. fT r? J: *4^0* ^^Sa4*<l^l*49l9k9tff'9t0'0,0'ff>0'0'ff>9k0'^0'0'0k0'0' ^-M:W:NNKM-4WO-4UtOU4= -=A*=J*01 'U= M= 4*O:`U: >:^W= M-. Os*4=N=-4U= *< f * * > *x ab. *r 13 of hapatocylas Focal aacroala of bapalocytaa foctl lymphoid infl llrat ions Hm l m* o t* Focal hypertrophy of lMptlocyl a n Nodular hyparplaaka of bapalocylaa C 1* 1 it Focal hlla duck hyperplasia II* pa lama r a ? e e s am S Cliolanglo-hapaloma ' Uettculum Cell Sarcoma a> 40 metaatalic . a - Miinniiry tumor, metastatic *1 ......... I'*< i HARTOLDMON0023559 AttOCLOR 1242 Tabulation of Individual Lirar Laaloaa in Rato 170 Saeri/lc* Intarral Dooo (Aaimal . La to l No. and Son ll o A a " s ? i il ii 11 iao'. i Lltor Lot loos If1o 2 u li fi 5J Vz . A n ia ou i <3 2 io U a ?m II sa =; 4 Mouth jc-oi 7403d | lOTppmi 722" 724" 744" 744" 748" 768T 771" j 77S" Ext" | 733" ' 765" j 7G4" 767" | 774" : 773" j 784" 789" 793" P P P P P u o Ca fc 3 ? ^Control' 6m; ii" | 12" j 21" j 3" j 33 34 35" I ; Ext" 46r 47" 49" ; 51" ; 52" > 54" j 59" ! 42" | Ii - o' I HARTOLDMON0023560 4 M onth C o n tro l 6JF t* + Modarmta la a a ve rity +*-- M a r had ia a a r a r ify P P ra a a n t. no gratia 3a nE c za S.? 3 jCytoplatfmlc v*coUiion of hapilocytfi Focal oacroaia of hapalocylaa Focal lymphoid laflltratlooa Focal hypertrophy of kapalooylaa Nt'tlolar hyparplaaia of liapalocylva Focal btla duel hyparplaaia s. I> 7P I? ~o ra it llapaloma Cliotanglo-Hupatoma Kalkulum Call Sarcuma mala Halle Mammary tumor, mala alalic *1 I., H|M- la a la, foial i & ii! I HARTOLDMON0023561 tiw wowm ioo NOtfMHOM.lUlNOil MIM ICPOtTTO MONSANTO COMPANY two - tea* chronic oral toxic:rr STVBT WITH AROC LOR HO IN AUDIO RATS hstopathological evaluation or ADDITIONAL LT'ER sections march m? 1ST NO. MI - OMTi 098 /'* HARTOLDMON0023562 Milhtil lO-TIfT M. Akl 099 NM nONTMl IOM % Nomueei. iujMO<a mm J*1/ 1. WTJ C|| J. t athiafcaa fk.D. MO S Ltod3ghBl*4. St. Unto. Waawirl 4J1M Dttr Dr. UtiMkiti ' 1 OTN*. MI-**72 Htotopalhatotltil inlamtUm ml AddMaaal Lhtr Itcta* Fraa >t ml a Two - Ym Qmlc Oral Toalctty Itud*. 11 Aroctor UU. , M ara --i--m*| hw hIM our rrrtiid Ithtmiry tap art dunl March 24, 1474. prepared to ccouctua with tha above mdy. Vary My yaure. fy. Z>. >Uo - jTc. <~ilaoitra JGC/M HARTOLDMON0023563 f EEPCftTTO Monsanto company TWO - YEAS OOIOKTC OEAL TOOnCITY STUDY WITH AJtOCLCl 1242 IN ALBBfO slaty KBTOPATHOLOGICAL EVALUATION or ADDITIONAL LIVE* SECTIONS MAJLCH 24, 14TJ 1ST NO. 441 04*72 L Tmtrodactloa At dlt reboott of Dr. Lnlukti of Ik JImmim CtBftir, oddltioool oocNom oI licar (tub o two r**' tkratlt oral toxicity itatfy o' Aroc lor 1242 la rati (1ST No. 422*072 90) ooro procotoad lata Hit icxtnoC ooctioct t: i ovotaotod by Ufht microtcoyy. Tbo following report protoato tfeo rotul.o r' tfclo rtady. HARTOLDMON0023564 IO.TIIT n. ta *a U that frtriMMlf Wral UM a < Tka J' _A 191 -.V mi Aa i 1242 to aa DBOSZSIAL BIO-TMT LABORATORIES. Rapart Praparad aad D. t. Gordot. D.V4I.. Pfc.D. Raport Approrad bp td U. L. KapUap J>. HARTOLDMON0023565 SmJmAtd tlO-TIST IBT Ko. 4Z2-fT2t If* HARTOLDMON0023566 ar -tfV ^ ( IO-TIST Wltfci i ti * eMt Wutt. Uxktar. D.T.M..M.S. DtpL, Am. C*tUf* Ttt PuX HARTOLDMON0023567 raawr Lhrar UaMM AtmIot Utt . - , ,u Bm - IUM . SiVv' Iditsa *w n T8 TB tmmiw ckm e/o 2/10 1/0 r*c*X Kyptitnykr o/e 0/10 0/0 Natal** HyftflkaU e/a 0/10 0/0 HtyMon* o/t 0/10 0/0 Ovctilu Kn*fyU<ia 0/1 0/10 1/0 CbalMfi** Kayatttn* e/t 0/10 0/0 Htytwwlhlir Nacroai* i/a 0/10 0/0 -. Cwnl 2/10 0/10 0/10 0/10 0/10 0/10 0/10 000318 -Cfc i HARTOLDMON0023568 TX TJX TJS Vtnilu 0/10 0/0 0/0 rcil 0/10 0/0 0/0 MoOaU* Hy^ryUiU 0/10 0/0 0/0 HtfUMM 0/10 0/0 o/o Ovcttlii C/10 0/0 0/0 atUnto-Hlyiwmi 0/10 0/0 0/0 Ht^uciUalu Xtemli 1/10 o/l 0/0 'C- Cwnl 1/10 0/10 0/10 0/10 0/10 0/10 1/10 $ HARTOLDMON0023569 PitM Unr UitH - Amid Ms- ; . u Urn* ImtUIm IM - ' Tiwlir CkMf TX 2/19 rtMiBffMtnykr 0/19 IMitar 9/19 0/10 DvcmIm MyyaryUat* 0/10 0/10 HtfitMtUtUr terU 0/10 TO 4/10 0/10 0/10 0/10 1/10 0/10 0/10 * : '"t m 0/0 1/0 0/0 0/0 0/* 0/0 0/10 T 10* C--trol 1/10 0/|0 0/10 0. 10 1/10 o/:o 0/10 HARTOLDMON0023570 ' >> ' " V' .t'.f '-* ! Prlauy Ltrmr Uiiwi * AtocIot BO Tmlait Skcrlflc* - Kata 107 Latloa VuMUr Gain* focal Hr7rtrfhr NoCular Hrp*r?Uia Hapaioma Doetalar HTfo^P1**u CboLaaflo-Hapatoota HapatoeoUular Nacroala t: T/ll 2/11 0/11 0/11 1/11 0/11 1/11 Sssat n nn 0/10 0/20 1/10 0/20 2/10 0/20 0/10 2/20 1/10 1/20 0/10 ` 1/20 1/10 o/:o - Control 1/23 0/23 1/21 0 21 S/21 u/21 l 23 a HARTOLDMON0023571 TOSA ! tw* : n o n 31 H Aiiillsi Ji.rJL * 3i?| Vocal mmli o( hapatocytaa Focal lymphoM laflltratlooa Vocal byparlroftlty W IW|MlM|ll No*ilf hyporpUaia ai Impatocytaa Vocal MU duel ItyparpMaU Ilo^atoma Cholaafla-hayatama laltculum Coll Sarcoma] mataalallc Mammary tumor, mataalallc To laniim la a 1.1, fu(*.il HARTOLDMON0023572 Aaoctoa . 1242 TiWUUoa mt l&tMdaal Uwt LUa to Bate ** 10 itcrillet Latarvml Da Aaimatj Ul No. htad 1 Sex h\ 1 & Uoath j C-I jlppm m. toiiM 1012" | 1013") 1014"| 1019"! 102*Fj 1027" 1029" 1029" 1030" ii*3i l1 la Urtr JUoloao fi }i {}H ii i*F I k I 'J:, l 2 -3* VJ it Ij If it I 4 LJL ;C-C 10413d ! lOppm 1042" I 1043" ! 1049" losor 1097" ibS9" 1039" i C.Ol 10713d lOCppra1072" 1073" 1073" ' 1096F 1097" | 1099" 1099" 1090" Control 90134 902" 903" 904" 909" our | 117" 119" 919" 920" ! HARTOLDMON0023573 HARTOLDMONOQ23574 *m m *ry him or, m tla iU ilc u r * 2 #trj? ^ 9A lg * f2 [1 *! BlHfUMlttltMhliM . -, (f klfMltyili rnl mcimU t( . . .:' : rCAl tyaaffcM, , . ^t ' kifMMftM . WIHWHIM f H * rml Ut*M :< [ \Itapktaaaifitelr >; > ' Pto- 'i': ' . * ^"4 4 t' Chl>nl| M*Nre. o u I(liculun Call Immim iMiutallt 1 Mammary tumor* IMUlUtiC HARTOLDMON0023575 HARTOLDMON0023576  t .. ' 1 4 c.ffl lOOppm CoatTol c * ir if m ** { * u |J *d *d *0 4 1 *d * * *d *u w a rir| Cytoplasmic wmtUllN of kiylN|tM FmI MtMl* oi rottiiiNfiwU . laOUttUMi Focal fcypotttpphy of hapatocytaa 8 NcMar krfitfbil* cf Mpatocytaa c ni Focal Mia 4act I *?**,-,, ; r it * `:U,; '**'? 'v * 9 1 Clmlaasto!>paterae <Vr . RdlcaHun Call klaHM nxItaUllc Mammary tumor, metastatic |\* \ |tl*t>l | ^ j.|. flM l| 0003 HARTOLDMON0023577 Grading 3rrtm * UUbmIIi Mvnrltf UiU in MvariTf -t-f Mndnmtn in imrily h++ iUfM in Mnrlty P Pntut, no gnda 000328 HARTOLDMON0023578 ' '*.** -V*. , v* ` ,.-?4 '*'** - \:* .~i CO Lciioo VacuoUr Chui| Focal Hypertrophy Nodular Hyperplasia Hepatoma LXictular Hyporplasia C ho lan gio- Hops loma Hepatocellular Necrosis- Primary Unr Utloai Aroclor 1242 Its Tarmiul iteriflci - Hat* T1 123 TU 12 Group 3 nn 12 3 Caitral 12 0/31 0/27 7/31 4/30 1/19 1/30 13/20*' S/21 9/20 1/23 0/24 0/31 0/27 2/31 1/30 2/19 3/30 1/20 2/21 0/20 0/23 0/24 0/31 0/27 0/31 1/30 0/19 2/30 2/20 S/21 0/20 1/23 0/24 0/31 0/27 0/31 0/30 1/19 0/30 0*20 0/21 2/20 0/23 0/24 0/31 4/27 3/31 1/30 0/19 3/30 20 2/21 3/20 S/21 b/24 0/31 0/27 0/31 0/30 0/19 0/30 0/20 0/20 1/20 0/23 0/24 1/31 0/27 1/31 1/30 0/19 1. 30 0/20 0/21 0/20 1/21 0/24 1 - First evaluation of original ilUtl 2 - Re*cviIuatian of original slides 1 = Evaluation of additional tiaauo Htlioai Fuur animals positiva on first evaluation, not on sscad. HARTOLDMON0023579 . -I LI 11 ,u<yj 1IO-T1IT iiMkMJb Frtnary Liver LmIou - AXOCLOft 1242 Tmtatl ImiIIIh hM " a< Original Slldao . Vacuolar Change Focal Hypertrophy Modular Hyperplasia Ductular Hyperplaaia Cholan gin-hepatoma Hepatoma C-l 0/27 0/27 0/27 4/27 0/27 0/27 C-2 1/1* 2/14 0/19 1/19 0/19 0/19 C-l S/21 2/21 S/21 2/21 0/21 0/21 1J6 Central 0/24 0/24 e/24 b/24 0/24 0/24 HARTOLDMON0023580 1 a U0-71ST JW wiGJim J4? '* j I .in ; ._ Ml f M .Ml f MM*l 11 u::i * *i: H 117 1t i HARTOLDMON0023581 I I ! rr I ;\ j I I 4 4 I I . it* j *r* t r. i: : i r 1111*011 811 HARTOLDMON0023582 H* * i i m rj nli.Miq pa H f* i-p***.*-!^ r>4 .....................! `i :i i n, n jr.5 is i >>* ' 1 ii 1 > W *' c: cj t. o c lQ i i k 'it - S& V:4 I l K HARTOLDMON0023583 t? rj c5 c ! I HARTOLDMON0023584 tu x x I B I T HARTOLDMON0023585 UX CEXTSAt DMEH FIU3 Toxicology Section/St. Louis ; (CO./CIVJD6?TiUOCATIO.NI 1^ , r ________ SPECIAL ITYPE OF REPORT) REPORT tCoU H-* 2 2z E-- cCO I--' >- iE----i t0o3 U H-. h2-. 22 W - to to o 2 > -- tj i-. Ci tj < >- fl. 'JZcu .X Cl. to c. a <-- t2o - 2 2 >* vi a s z a >-- ai ^ vi E- C 2Z C2- C 2>4) 22 >< 2 < to c-- < C-3* 2 at . 2 a: c-- < 0 to oao 2< S t>>u- 2Z <Z.t-e>i u aat t>o-. 2u to Ca: 2 >2 vai: ^< lz-H ac- o .. X lit 5 p! <P ro oo(NJ JW ^ . REPORTNO.: MS L-2 00 3 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: A REVIEW AND EVALUATION OF CARCINOGENICITY STUDIES IN MICE AND RATS AND MUTAGENICITY STUDIES WITH POLYCHLORINATED BIPHENYLS AUTHORS: George J. Levinskas, PH.D. A3STRACT: This is a review and evaluation of studies which deal with the potential carcinogenicity and mutagenicity of polychlorinated biphenyls (PC3s). It is subdivided into 4 sections: Chronic Rodent Studies, Metabolism Studies, Co -Carcinogenesis Studies and Mutagenicity Studies. A brief summary of Epidemiology Studies is added to complete coverage of the issue of carcinogenicity. R-ioasiat SCM 019651 00033^ HARTOLDMON0023586 COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - DMEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H. Craddock, A2SA 8 - G.J. Levinskas, G2WF 9 - J.G. Nassif, E2.ND . ' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: _____________ REPORTS LIBRARY, R2C______________ If you transfer it to anyone else, please let your librarian know, so the records can be changed. R-iaai(C) (Rev. l/ao) SCH 019652 000336 HARTOLDMONOQ23587 INTRODUCTION This is a review and evaluation of studies which deal with the potential carcinogenicity and mutagenicity of polychlorinated biphenyls (PC3s). It is subdivided into 4 sections: Chronic Rodent Studies, Metabolism Studies, Co-Carcinogenesis Studies and Mutagenicity Studies. A brief summary of Epidemiology Studies is added to complete coverage of the issue of carcinogenicity. This review does not discuss the effects of impurities or contaminants, particularly polychlorinated dibenzofurans (PCDF), which are reported to be present in some PCB mixtures (Brinkman and deKok, 1980). The presence and amounts of such impurities have not been specified in the materials used in many studies. Thus, attempts to apportion the observed biological effects between impurities and PCBs would only add further " conjecture to a subject which currently is rife with speculation. By contrast, specific chemical and trade names have been used to identify materials that were studied instead of the all encompassing term PCBs. This was deemed necessary because there are too many one-sided generali zations in the literature. Every adverse finding is, by implication at least, extrapolated to the entire class of materials designated as PCBs. Conversely, every report which has failed to find an adverse effect is careful to stipulate that it relates only to the substance studied. Even more difficult to contend with are misleading references to adverse findings which are not substantiated by the actual publications referred to. An example of this occurred in a discussion of in vitro metabolism studies with liver microsomal enzymes which stated that the formation of PCBmacromolecular adducts had been demonstrated. Among the references ` 1` SCM 019653 000337 HARTOLDMON0023588 cited was a paper entitled "The in vitro binding of 2,2',5,5'-tetrachicrobiphenyl metabolites to rat liver microsomal proteins". The latter paper clearly states "There is no clear evidence in the data obtained from the work reported here to substantiate covalent binding; however, it is also not "possible to exclude the nondialyzable radioactivity as being covalently bound". (Hargraves and Allen, 1979). To emphasize the point that dose is important in evaluating safety, test exposure conditions have been described so that the reader can compare them to ambient exposure levels encountered in. the literature and else where. Hoopingarner, et al. (1972) in their studies with Aroclor 1254 on cultured human lymphocytes are among the few authors who acknowledged that "The toxic dose of the chemical was several times greater than is usually found biologically". They used concentrations of 100 ppm of Aroclor 1254 in their studies discussed under Mutagenicity. Chronic Rodent Studies Difficulties in comparing and assessing different studies are compounded not only by problems of histopathologic diagnosis but also by variations in experimental design and animal strain differences. Animal feeding studies cited in the literature as evidence for the carcinogenicity of PCBs and related studies of similar duration have been tabulated for mice in Table 1 and for rats in Table 2. Studies have been grouped by the approximate weight percent of chlorine in the materials tested to facilitate comparisons between products from different manufacturers with the same chlorine content. 2 SCM 019654 0Q033F HARTOLDMON0023589 Mouse studies in Table 1 were conducted with high dietary levels of PCBs. Kimbrough and Linder (1974) reported a 52% mortality for mice fed 300 ppm of Aroclor L254 for 6 months and then held an additional 5 months. During that interval, control mice had a 32% mortality. This appears to be quite high for control mice about 1 year of age. In a report discussed under Co-carcinogenesis, Roller (1977) studied mice injected with Aroclor products and Moloney leukemia virus. Some data from his study, although not shown in Table 1, are quite similar to those of Kimbrough and Linder (1974). Roller (1977) fed diets containing Aroclor 1221, 1242, or 1254 to groups of 25 Balb/c male mice for 6 months. After that time, some mice were sacrificed for examination and others were returned to a control diet for another 3 months and then sacrificed. Dietary levels of each Aroclor were 375, 37.5, or 3.75 ppm. The only feeding regimen that was toxic was 375 ppm of Aroclor 1254. Only 8 mice of that group survived for 6' months, giving a mortality of 68%. Other authors cited in Table 1 (Ito, et al. , 1973a,b and Nagasaki, et al. , 1972, 1974, 1975) did not indicate how many of the mice in their studies died when fed 500 ppm of Kanechlor 500, which has a chlorine content similar to Aroclor 1254. Roller (1977) reported that "PCBs did not produce hepatic neoplasia".There was marked liver injury in mice fed 375 ppm of Aroclor 1254, mild liver injury which persisted through the 3-month recovery period at 37.5 ppm, and no hepatic lesions at 3.75 ppm. Moderate liver injury was produced by 375 ppm of Aroclor 1242, but this regressed and no hepatic lesions were seen 3 months after mice were returned to a control diet. Other dietary levels of Aroclor 1242 and all levels of 1221 did not produce liver changes. Highly significant mean liver weight increases after 6 months of feeding occurred in all Aroclor 1254 groups and in the group fed 375 ppm of Aroclor 1242. Three months after mice were placed on a -3 SCH 019655 onorw HARTOLDMON0023590 control diet, mean liver weights of each group had decreased. The decreases were highly significant for the 37.5 ppm Aroclor 1254 and the 375 ppm Aroclor 1242 groups. These observations are generally consistent with those of other investigators which show regression of lesions when PCQ dosing is ended, the degree of regression depending upon the duration of the recovery period. . Data for mice reported earlier by Nagasaki, et al. (1972) appear to have been included in the later publications of Ito, et al. (1973a,b). It is interesting to note that the tumors were called hepatomas in the former publication and were labeled well-differentiated hepatocellular carcinomas in the latter. Kimbrough and Linder (1974) apparently also felt that these same data on mice were reported in these 2 papers as they reference the . production of hepatomas in male dd mice to the later publication by Ito, . et al. (1973b). Similar terminology troubles beset the 1974 and 1975 Nagasaki publications. Data from the later publication list 9/17 males and 4/17 females fed Kanechlor 500 as having liver tumors. Table 1 shows data from the earlier publication; among males, 9/17 had nodular hyperplasia and 7/17 had hepatocellular carcinoma while the female incidence of 4/17 was described as nodular hyperplasia. The hepatomas described by Kimbrough and Linder (1974) were later referred to as "neo plastic nodules (hematomas [sic], hyperplastic nodules)" by Kimbrough, et al. (1978). The use of the term hepatoma has created confusion. With respect to terminology of tumors in the mouse liver, "Hepatoma is a collective term used to describe the progressive stages of tumour development from the lesion called "hyperplastic nodule" or "simple hyperplastic growth" to the morphologically and biologically malignant neoplasms" (Turusov and Takayama, 1979). Nevertheless, whatever '4* SCH 019656 000340 HARTOLDMON0023591 terminology is used to describe them, tumors were attributed only to PCBs containing 52-54 percent chlorine. Lower doses of the 52-54 percent chlorine materials, as well as lower chlorinated materials, were not described as tumorigenic. Table 2 summarizes results of rat studies with PCBs. As in the mouse studies, only some studies with materials having a chlorine content of 52-54 percent, or higher, were reported to produce carcinomas in the Livers of rats, and even for those materials this was not a consistent, reproducible observation. Odashima (1976) reported the results from a series of different tests used to evaluate potential carcinogenicity of several compounds, including PCBs. Two tests are of sufficient interest to mention here. One is the transplacental method in which rats were treated for 3 days during pregnancy. These were days 15, 17, and 19 or 14, 16, and 18, depending on the strain used. The total dose was approximately the maximum one that did not cause abortion or early death of the weanlings. In the other, the newborn method, pups were dosed subcutaneously on days 1, 8, 15, and 22 after birth. The maximum dose was one that did not cause early death of over 20% of the animals. The subsequent observation period in each test was limited to one year after birth. For both Kanechlor 300 and Kanechlor 500, the transplacental test was scored negative and the newborn one as equivocal. Both of these tests gave positive results with some known carcinogens such as 4-aminobiphenyl, benzo [a] pyrene, butyl-nitrosourea and N-methyl-N-nitrosourea. SCM 01 Qf> 57 000^41 HARTOLDMON0023592 Objective appraisal of chronic rodent feeding- studies is difficult. Some .of the rodent studies in Table 1 and Table 2 are of the type used to detect potent carcinogens. They use relatively few animals, high doses of the test material, and have a relatively short duration. Studies of that nature have been included in Tables 1 and 2 to illustrate the various diagnoses of the rodent hepatic lesions observed after dosing with PCBs and to aid in the stepwise analysis of the results of all the reported carcinogenicity studies. In any short-term test with a few animals, the chance occurrence of some tumors is a possibility which must be considered. The probability that an event is a chance occurrence is decreased if it is repeatable. Many of the studies in Table 1 did not produce tumors. Those which were reported to ' produce hepatocellular carcinoma apparently were also reported in other publications as producing nodular hyperplasia, or tumors, or hepatomas. Since terminology of tumors and the use of those terms have subjective, i.e., judgemental, elements it may not be valid to make direct comparisons between different studies on the basis of terminology alone. The rodent studies which were of less than lifetime duration raise a question as to whether or not cancers would have occurred had the feeding periods been extended. That question is speculative, and it cannot be answered from the present data. Nevertheless, negative studies even if they are of relatively short duration are part of the overall evidence which has to be considered. As a first step in the analysis of the data, the evidence shows that at a minimum, PCBs are not potent carcinogens to rodents because they do not produce cancers when tested in studies designed to detect potent carcinogens. *6* SCM 019658 AAno-l?. HARTOLDMON0023593 With respect to further analysis of che mouse studies, greater significance can be attached to the data of Kimbrough and Linder (1974) because their study had a larger number of animals, a longer duration, and a relatively high dietary level of PCS. In those respects, it more closely resembles conventional cancer studies. They did not observe any hepatocellular carcinomas. Thus, it can reasonably be concluded that PCBs are not carcinogenic to mice. Review of the rat data in Table 2 shows that there are 2 studies on PCBs with an average chlorine content of 40% (Levinskas, 1SS1 and Weltman and Norback, 1979) and 3 on PCBs wich an average chlorine content of 52-54% (Levinskas, 1981; NCI, 1973; Wasserman, et al. , 1978). Results of those studies are consistent with respect to the absence of hepatocellular carcinomas. This consistency reasonably suggests a conclusion that PCBs ' wich those chlorine contencs are not carcinogenic. Of the 3 studies wich PCBs having an average chlorine content of 60%, one reported hepatocellular carcinomas (Kimbrough, et al. 1975) and 2 did not (Levinskas, 1981 and Weltman and Norback, 1978). Since Kimbrough, et al. (1975) and Levinskas (1981) both used Lot No. AK-3 of Aroclor 1260, che different conclusions they reached are not related to differences in the test material. In addition to the use of a different strain of rat, Kimbrough, et al. (1975) used a different histologic diagnostic criteria. Kimbrough, et al. (1975) used the criteria for classification of specific hepacoceLlular lesions in rats developed at a National Cancer Institute Workshop (Squire and Levitt, 1975). That workshop recommended that the term "neoplastic nodules" replace so-called "hyperplastic nodules" because "Such nodules are proliferative lesions and -7- SCM 019659 000.143 HARTOLDMON0023594 are known co be induced by carcinogens and, ac che least, they indicate an increased probabiity for the development of hepatocellular carcinoma" (Squire and Levitt, 1975). However, Pitot, et al. (1978) in a discussion of stages in the progression of hepatocarcinogenesis in rat liver have noted that "The demonstration of carcinoma cells that appeared to arise within the nodules was also reported (13)1, suggesting the nodule was a precursor to the malignant neoplasm. On the other hand, as was shown by Farber and others, the vast majority of the regenerating or hyper plastic nodules disappeared on removing the animals from the carcinogenic diet. Thus, despite the more recent suggestion that these nodules be termed "neoplastic nodules" (14)2, it is difficult to understand a precursor relationship of the nodule to carcinomas if the existence of the putative precursor is so transient." Further, the use of those criteria for classifying experimental hepatic lesions was considered and rejected by Kimura, et al. (1976) in their studies on the co-carcinogenesis of Kanechlor 400 and 3'-methyl-4-dimethylaminoazobenzene. There is concern' that a presently benign lesion might at some future date or under other circumstances be transformed into a malignant lesion. Kimbrough (1979) apparently had that concern when she stated "Although it has been shown many times that the neoplastic nodules are part of the carcinogenic response they are not always included in the statistical evaluation of bioassays, which may lead to erroneously interpreted results, particularly when they are classified as hyperplastic nodules or "nodular hyperplasia" and when the number of animals studied was small (Carcinogenesis Testing Program, 1977)".3 A similar concern appears to have been behind the statement in a recent review (Anon., 1981) that "hexachlorobiphenyl administered to groups of 50 male and 50 female 8 SCM 019660 000344 HARTOLDMON0023595 Sprague Dawley rats ac dietary levels of 0 and 100 ppm for 105 weeks was carcinogenic in female rats, producing an increased incidence of liver hepatocellular carcinomas among the dosed animals (Norback and Weltman, 1980. Personal communication)1'. Contact with Dr. Norback (Ribelin, 1981) revealed that her data were available only as an abstract (Weltman and Norback, 1979), and that the abstract and her oral presentation of the data referred to the lesions as neoplastic nodules, i.e., not distinctly tumorous. These 2 examples illustrate the difficulty in establishing that cancer is not present, and they strongly suggest that the different findings reported by various researchers are a reflection of their orientation, training, and philosophical perspective. The latter is defined as the difficulty in separating what is actually being observed under the microscope, from a concern over what it might have become if the animals had lived longer. Since only an abstract has been published, details on the studies by Weltman and Norback (1979) are limited . Their studies are of particular interest because they observed the sequential development of liver changes over a 2-year period. They noted that hexachlorobiphenyl was more toxic than tetrachlorobiphenyl, and that the more toxic, more highly chlorinated hexachlorobiphenyl produced neoplastic nodules only. Again, the weight of the evidence leads to a reasonable conclusion that the carcinogenicity of biphenyls with an average chlorine content of 60% has not been established. 9 " . SCH 019661 O 00.145 HARTOLDMON0023596 Overall, one can, surmise chat the inability to resolve the crucial issue of whether or not a lesion is neoplastic in character was one of the factors which led to the following1 statement from a recent Surgeon General's report: "Science, and society have not yet arrived at a final consensus on the detinition of a carcinogen either in the human population or in experimental animals" (DHHS, 1980). Metabolism Studies Several reviews (Goldstein, 1980; IARC, 1978; Matthews and Kato, 1979; Roberts, et al. , 1978; and Safe, 1980) discuss the metabolism of specific isomers as well as mixtures of PCBs. While there are some exceptions, the following general conclusions can be drawn. Both the degree of chlorination and the positions of the chlorine substituents determine the ease with which PCBs are metabolized. In general, the lower chlorinated ones are metabolized and excreted more readily while the more highly chlorinated materials are stored in fat. The process of metabolism converts the fat soluble PCS into a water soluble hydroxylated derivative which can be excreted in the urine. This metabolism and excretion appears to occur via arene oxide intermediates, and the carcinogenicity of some compounds has been attributed to formation of arene oxide interme diates and their binding to subcellular macromolecules. Chlorination at the 4,4' position blocks the metabolism and excretion of PCBs as illustrated below. SCM 019662 0Q0ri4G HARTOLDMON0023597 Studies in rats (Hansell, et al. , 1977) and in mice (Morales and Matthews. 1978, 1979) are of interest because they report the metabolism of isomers which were fed to rats for 2 years by Weitman and Norback (1979). Hansell. et al. (1977) gave groups of male rats single intraperitoneal injections of 0.2 nmoles/kg of 2,2',5,5'-tetra- or 2,2\4,4',5,5'hexachlorobiphenyl (TCB and HCB, respectively). They measured changes in hepatic mixed function oxidases and the persistence of the PCB in the livers of animals killed at selected intervals for 35 days after dosing. TCB produced a transient, but significant increase in O-demethylase activity only at day 3 while HCB produced significant increases in O-demethylase and aniline hydroxylase activities within 24-48 hours. Induced enzyme activities by HCB peaked at 4-6 times control activity during days 7-14 and were about 3 times control activity at the end of 35 days. Even though equimolar amounts of each isomer were given, liver ' residues of HCB one day after dosing were about 7 times higher than those of TCB. HCB residues decreased relatively slowly with time while TCB residues were more rapidly eliminated and had almost returned to control values by 35 days. Livers from HCB treated rats had centrolobular necrosis at day 35. They also were significantly increased in size, showed increased amounts of smooth endoplasmic reticulum (SER), and appeared to contain increased numbers of microbodies and reduced amounts of rough endoplasmic reticulum throughout the 35 day interval. By contrast, TCB treated livers were similar to control ones except for occasionally increased aggregates of SER on days 3 and 4. Hansell, et al. (1977) stated "It would be interesting to speculate that.. .2,4,5,2',4',5'hexachlorobiphenyl undergoes direct hydroxylation whereas 2,5,2' ,5'tetrachlorobiphenyl undergoes bio transformation via the arene oxide intermediate, thereby accounting for the different slope for the - 11 - . SCM 019663 000.147 HARTOLDMON0023598 elimination curve, of the latter compound." Thus, the metabolism (Hansell, et al. 1977) and feeding1 (Weltman and Norback, 1979) study results lead to somewhat different conclusions. The more potent enzyme inducing, less readily excreted HC3 which appears to resist hydroxylation produces neoplastic nodules in rat livers. By contrast, the more readily excreted TC3, which apparently is excreted via an arene oxide intermediate, does not produce tumors when fed to rats for 2 years. Covalent binding to cellular macromolecules also appears to be greater for the more readily metabolized PCB isomers. Morales and Matthews (1978, 1979) compared the covalent binding of 2 hexachlorobiphenyls; the more readily metabolized 2,2',3,3',6,6'-hexachlorobiphenyl (2,3,6-isomer) and the more slowly metabolized 2,2',4,4',5 ,S'-hexachlorobiphenyl (2,4,5-isomer). Each PCB, with a radiocarbon label, was given orally to groups of mice at a dosage of 7.28 mg/kg on each of five successive days. Animals were killed 1, 5, and 8 days later. The concentration of each PCB was determined in liver, muscle, and kidney. All tissues had consistently higher concentrations of the less readily metabolized 2,4,5-isomer. The more readily metabolized 2,3,6-isomer showed a consistently greater binding to purified macromolecules. The binding was at least one order of magnitude greater than that seen with the 2,4,5-isomer. Results of animal feeding studies with the 2,3,6-isomer would be of particular interest to determine the degree of correlation, if any, between the carcinogenicity of this isomer and its covalent binding to cellular macromolecules. Taken as a whole, metabolism studies suggest that if PCBs are carcinogenic, it should be those PCBs which are more readily metabolized and excreted, i.e., the lower chlorinated materials. This is in sharp - 12 - SCH 019664 000.74/9 HARTOLDMON0023599 contrast to results on animal studies in which questions of carcinogenicity arise regarding higher chlorinated materials only. On balance, metabolism studies on the formation of arene oxide intermediates would lead one to expect lower chlorinated materials to show a more pronounced carcinogenic response in animals. Conclusions drawn from metabolism studies are not supported by animal feeding studies. Co-Carcinogenesis Studies The position and degree of chlorination of PCBs also are important in inducing microsomal mixed function monooxygenases (Goldstein, 1980 and Yoshiraura, et al. . 1979). Such induction of microsomal monooxygenases could alter the metabolism of exogenous and endogenous substances in the body, as reported in a variety of studies which have been conducted to determine whether PCBs might be cocarcinogens. These are summarized in Table 3 and discussed in greater detail below. Uchiyama and Chiba (1974) inserted 20-methylcholanthrene impregnated threads into the uteri of virgin mice and fed them diets containing up to 100 ppm of Kanechlor 400 or DDT. Animals were killed at various times and the cervical epithelium was examined for cancerous changes. Kanechlor 400 dosed mice showed no significant changes as compared to the controls, but those receiving 100 ppm of DDT "showed a remarkable tendency towards the induction of cancer." Diets containing benzene hexachloride (BHC) isomers and Kanechlor 500 separately and in various combinations were fed to male mice by Ito, et al. (1973a,b). Concentrations of the BHC isomers ranged from 250 ppm to - 13 SCH 019665 0^0.^ 3 HARTOLDMON0023600 50 ppm. The amount of Kanechlor 500 was 250 ppm or 100 ppm. Test groups consisted of 20 to 30 animals. When fed alone, only a-BHC at 250 ppm produced a high incidence of noduiar hyperplasia and a moderate incidence of hepacocellular carcinoma. A diet containing 250 ppm each of a-BHC and Kanechlor 500 increased the number of hepatocellular carcinomas. In comparison, combinations of 100 or 50 ppm of a-BHC and 250 ppm of Kanechlor 500 yielded a moderate incidence of nodular hyperplasia and produced only a few hepatocellular carcinomas. There were a few nodular hyperplasias in mice fed 100 ppm each of a-BHC and Kanechlor 500. A mixture of 50 ppm a-BHC and 100 ppm of Kanechlor 500 was without effect. Diets containing 250 ppm or 100 ppm of p-BHC and 250 ppm of Kanechlor 500 produced both nodular hyperplasia and hepatocellular carcinomas at a lower incidence than that seen with comparable diets of a-BHC. No liver nodules were seen with 50 ppm of p-BHC and 250 ppm of Kanechlor 500 or with 100 ppm of each. y-BHC and Kanechlor 500 did not produce liver nodules either singly or in combination. Nagasaki, et al. (1974, 1975) reported a similar series of experiments except that Kanechlor 400 was included. The same concentrations of the PCBs, 250 and 100 ppm, were used. Their test groups consisted of 20 to 38 male mice. For a-BHC and Kanechlor 500, they presented essentially the same data as Ito, et al. (1973a,b). While Kanechlor 400 did not produce liver nodules when fed alone at 250 ppm, there was an increase in the incidence of hepatocellular carcinomas when it was fed in combination with 250 ppm of a-BHC. The increase was similar to that reported by Ito, et al. (1973a,b) for a-BHC and Kanechlor 500. A combination of 100 ppm of a-BHC and 250 ppm of Kanechlor 400 produced only a few nodular hyperplasias and 100 ppm of each in the diet did not induce any liver nodules. - 14 SCM 019666 HARTOLDMON0023601 The effaces of PCBs on cumor induction by diethylnitrosamine (DEN) have been studied extensively. Male rats were given 25 ppm of DEN in their drinking water and 500 ppm of Kanechlor 500 in their diet concurrently for 20 weeks, returned to a control diet for 4 weeks, and then killed (Makiura. et al. 1974). Neither liver tumors nor nodular hyperplasia were seen, even though rats receiving the same DEN treatment alone had a 92% incidence of liver cancer. The livers of rats treated with Kanechlor 500 only also had no tumors. When administered after DEN, Kanechlor 500 markedly enhanced liver tumor production as reported in a series of papers by Nishizumi (1976, 1979a, 1979b). He described studies in which male rats were given 50 ppm DEN in drinking water for 2 to 10 weeks, after which they were intubated twice weekly with a com oil solution of Kanechlor 500 for 6 to 12 weeks. Kanechlor 500 doses were either 0.2 ml of a 5% or 0.1 ml of a 10% solution. Since experiments were terminated at 20 or 52 weeks after dosing, some animals received untreated diets prior to sacrifice. Only one paper (Nishizumi, 1976) contained results for animals dosed only with Kanechlor 500 alone. That treatment produced enlargement of the livers of rats but no neoplastic lesions. After pre treatment with DEN, however, hepatocarcinogenesis was enhanced as evidenced by the earlier appearance and a significant increase in the number of tumors. A similar dosing pattern was used by Preston, et al. (1981). Male rats were given drinking water containing 66 pg DEN/ml (66 ppm) for 5 weeks after which they were fed for 18 weeks on a control diet or one containing 100 ppm of either of 2 Aroclor 1254 diets. One was Aroclor 1254 as received. The other was an Aroclor 1254 which the authors claimed to have purified by removing polychlorinated dibenzofuran (PCDF) impurities by adsorption onto and subsequent elution from activated Florisil. No analysis was made of Aroclor 1254 as received for " 15 " . SCM 019667 000332 HARTOLDMON0023602 PCDF. The authors reported recovery of PCDF at a level comparable to 30 pg of tetrachlorodibenzofuran (TCDF) per 10 g of Aroclor 1254. TCDF was used as a positive control and a standard for quantification for the purification process. Other animals received one of the 2 Aroclor 1254 diets only during the latter 18-week interval. ' No evidence of hepatic tumor formation was seen in control animals or those receiving 100 ppm of either Aroclor 1254 diet. However, using the diagnostic criteria of Squire and Levitt (1975), they reported that both Aroclor 1254 diets resulted in significantly greater incidences of hepatocellular carcinomas in rats pretreated with DEN as compared to those dosed with DEN alone. Thus, they concluded that the hepatic tumor-promoting ability appears to reside in Aroclor 1254 itself. As the authors of this paper pointed out, it cannot be concluded that PCDFs are not promoters of hepatocarcinogenesis since appropriate studies have not been done on PCDFs alone. Similarly, their findings do not invalidate the hypothesis that some of the other effects reported for some PCBs may have been due to PCDF impurities. The dependence of the inhibition or promotion effect of PCBs on DEN-induced tumors on the dosing sequence was illustrated in a different manner by Nishizumi (1980). Pregnant female rats were given oral doses of 200 mg/kg or 50 mg/kg Kanechlor 500 on days 5, 10, and 15 of gestation and were allowed to deliver their pups. At 28 days of age, pups were given 50 ppm of DEN in their drinking water for 5 weeks. Groups of these pups were sacrificed at 16, 20, and 24 weeks after the start of DEN dosing and their livers were examined. The number of liver tumors in the offspring from Kanechlor 500 treated dams was significantly decreased as compared to the controls. The decreases were more pronounced in males. This decrease in DEN-induced tumors apparently resulted from induction of - 16 SCH 019668 HARTOLDMON0023603 microsomal enzymes in the livers of the pups. Those livers contained Kanechlor 500 residues and electron microscopy showed an increase in the smooth endoplasmic reticulum of hepatic cells. Rainbow trout were fed 6 ppb of aflatoxin Bi (AFB!), or 100 ppm of Aroclor 1254, or a combination of both for a year (Hendricks, et al. , 1977). At intervals during' the year, some fish were killed. The remainder were killed at the end of that time. All livers were examined grossly and microscopically. There were no tumors in those fed a control diet or Aroclor 1254 alone, and growth was not affected by 100 ppm of Aroclor 1254. The number of tumor-bearing fish in the group fed AFB t, plus Aroclor 1254 was significantly reduced (to less than one-half) when compared to those fed AFB!, alone. There also were fewer tumors per liver and the tumors were smaller in those fed the mixture. ' In the studies cited earlier, Makiura, et al. (1974) also fed combinations of 500 ppm Kanechlor 500 with 300 ppm of 3'-methyl-4-dimethyIaminoazobenzene (3'-Me-DAB) or 150 ppm of N-2-fluorenylacetamide (2-FAA). Results similar to those when PCBs were fed concurrently with DEN were obtained. Kanechlor 500 reduced the incidence of liver tumors to zero from 65% for those treated with 3'-Me-DAB and from 54% for those treated with 2-FAA. However, the effect of PCBs on the tumorigenicity of 3'-Me-DAB appears to depend on the dosing sequence as shown for DEN. Kimura, et al. (1976) used different sequences to feed groups of rats diets containing 400 ppm of Kanechlor 400 for six months and 600 ppm of 3'-Me-DAB for 2 months. Some received Kanechlor 400 alone and some only 3'-Me-DAB. Others received one diet, then the other, after a 2-month interval on control diet. A final group received Kanechlor 400 for - 17 - SCM 019669 HARTOLDMON0023604 4 months and both materials for another 2 months. No hepacocarcinomas were produced by Kanechlor 400 alone or when it was given before or during the overlap with 3'-Me-DAB. No hepatocarcinomas occurred in control animals. The incidence of liver cancer rose from 13% in those receiving 3'-Me-D AB alone to 64% in the group which received Kanechlor 400 after 3'-Me-DAB. The inhibitory effect of Kanechlor 500 on rat liver tumors was evident when rats were dosed with two carcinogens (Makiura, et al., 1974). Combinations of 3'-Me-DAB and DEN or 2-FAA and DEN were administered with and without Kanechlor 500 at the concentrations stated earlier. The liver cancer incidence fell from 92% to 8% when Kanechlor 500 was given to rats dosed with 3'-Me-DAB and DEN. It went to zero from 82% for those receiving 2-FAA and DEN. Nishizumi (1979a,b) studied the effects of various combinations of DDT and sodium phenobarbetal (SPB), in the absence and in the presence of Kanechlor 500, on the induction of rat liver hepatocellular carcinoma by DEN. Pretreatment of rats with DEN followed by DDT, by SPB, or by a combination of both produced low incidences of liver cancer while DEN pretreatment alone did not induce cancer. When Kanechlor 500 was included with each of the preceding dosing regimens, there was a marked increase in liver cancers. Increases resulting from joint administration of compounds after pretreatment with DEN were lower than those observed for DEN followed by Kanechlor 500 alone discussed earlier. Ito, et al. (1978) fed diets containing 200 ppm of 2-FAA to male rats for two weeks and then a diet containing 1000 ppm of an unspecified Kanechlor - 18 SCM 019670 000354 HARTOLDMON0023605 mixture for 8 weeks. Partial hepatectomies (PH) were performed on some rats during- the third week of the study. Feeding of 2-FAA was without effect, but that diet plus PH produced a few hyperplastic nodules in the Liver. Treatment with both diets caused an even greater incidence of hyperplastic nodules and both diets combined with PH produced a marked rise in the number of liver nodules. DiGiovanni, et al. (1977) and Berry, et al. (1978, 1979) studied Aroclor 1254 in a two-stage mouse skin carcinogenesis assay to see if it was a tumor initiator or promoter. The shaved skin of female mice was dosed with Aroclor 1254 at a level of 100 pg or 625 pg per mouse. This was applied alone as an initiator or from 5 minutes to 72 hours before initiation with 7,12-dimethylbenz(a]anchracene (DMBA). One week later, mice received twice weekly applications of 5 pg of the phorbol diester' promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) for 32 weeks. Animals were observed for both papillomas and carcinomas. Aroclor 1254 alone produced a few papillomas. The authors of these reports apparently faced a dilemma- common to scientists, i.e., how much significance should be attached to the observation of a low incidence finding which may or may not have a causal relationship to treatment. On the basis of these few papillomas, Aroclor 1254 was called a "weak tumor initiator" (DiGiovanni, 1977). Later, it was described as possessing "little or no tumor-initiating properties" (Berry, et al. , 1979). While Aroclor 1254 had a negligible effect on tumor induction when given 5 minutes before an initiating dose of DMBA, it markedly inhibited tumor induction when given 18 to 72 hours before DMBA. In other studies, an initiating dose of 200 nmole of DMBA was applied to the shaved backs of mice. After one week, 100 pg of Aroclor 1254 was applied twice weekly for 30 weeks. Aroclor 1254 failed to - 19 - SC* 019671 000.755 t, HARTOLDMON0023606 promote tumors while 0.2 yg doses of TPA applied in a similar manner induced an average of 8 papillomas per mouse. The authors concluded that Aroclor 1254 possessed little or no tumor-initiating or tumor-promoting properties. . The transplantabilicy and growch of Walker 256 carcinosarcoma in rats was inhibited by Aroclor 1254 (Kerkvliet and Kimeldorf, 1977a,b). Diets . containing up to 800 ppm of Aroclor 1254 were fed to rats of both sexes for 30 days, after which they were given incramuscular injections of Walker tumor cells. Nine days later, during which time they continued to receive Aroclor 1254 in their diets, animals were killed and the tumors were dissected out and weighed. Mean tumor weights of all Aroclor 1254 fed groups were significantly reduced as compared to their sex-matched controls, and the reductions were dose-related to the dietary level of ' Aroclor 1254. Body weight gain was depressed in males receiving 400 ppm or more of Aroclor 1254. Females showed reduced body weight at 100 ppm, the lowest level fed to that sex. Intraperitoneal injections of 50 to 200 mg/kg every other day for 14 days after injection of a small inoculum of Walker 256 cells (103) inhibited both the development and growth of the Walker tumor. The number of tumor takes and the size of the tumors were reduced and the tumor latency period was increased. Body weight gain was reduced only at the 200 mg/kg dosage. Alternate day intraperitoneal injections of 100 mg/kg and 200 mg/kg of Aroclor 1254 after a large inoculum of Walker 256 cells (107) were continued for 60 days. Control animals receiving tumor cells only had 100% mortality by day 20 with a mean survival time of 13.5 days. Mean survival times were significantly increased to 17.7 days and 18.9 days for animals dosed with 100 mg/kg and 200 mg/kg of Aroclor 1254. A few animals survived to 60 " 20 " SCM 019672 00Q35C HARTOLDMON0023607 days, and four receiving Che higher dosage of Aroclor 1254 showed cocal regression of their bilateral tumors. Tumor growth rates in Aroclor-treated animals were significantly reduced. Again, body weighc gain was depressed only at Che 200 mg/kg dosage. An experiment was undertaken to determine if there was an optimum time period for the antitumor effect of Aroclor 1254. InCraperitoneal injections of 100 mg/kg of Aroclor 1254 were given on 5 consecutive days before tumor inoculacion, on 5 days after inoculation, and daily from 5 days before until 10 days after inoculacion. This study was ended 14 days after Che initiating tumor inoculum was given. While all dosing schedules reduced tumor growth, there were variations in the responses. The greatest inhibition of tumor growth resulted from dosing animals for 15 days. Almost equally effective were precreatment and treatment starting with inoculation of the Walker cells. The delayed treatment starting after 5 days was least effective as Che tumor had become established. While early treatment reduced tumor growth, it was less effective in preventing metastases and death of the animals. - Kerkvliet and Roller (1980) offered groups of male mice diets containing 10, 100, or 500 ppm of Aroclor 1254 for 15 weeks prior to injecting them with MSB, an established tissue culture cell line derived from Moloney sarcoma virus. They measured tumor growth and cell-mediated toxicity over the next 19 days. A dietary level of 500 ppm of Aroclor 1254 resulted in marked weight loss and deaths. This was reduced to 250 ppm at 11 weeks, and a few weeks later surviving animals were placed on a control diet. They reported that Aroclor 1254 pretreacment enhanced the growth rate of the MSB tumor and inhibited or delayed the development of cellular immunity against the tumor. SCM 019673 000357 HARTOLDMON0023608 KoUer (1977) fed groups of mice diets containing 3.75, 37.5, or 375 ppm of Arocior 1221, Aroclor 1242, or Aroclor 1254 for 6 months. About 2 weeks after being placed on test diets, some mice in each group were inoculated intraperitoneally with Moloney leukemia virus. None of the dietary levels of -any Aroclor affected, i.e., did not promote or induce, the oncogenesis of Moloney leukemia virus. Since PCBs are enzyme inducers, their biological effects resemble those of other enzyme inducers. Peraino, et al. (1978) noted that phenobarbital had a specific tumorigenic enhancing effect that was restricted to the liver. With the exception of the mouse skin painting studies and those in which tumor cells or a virus were injected, all of the studies in Table 3 were concerned with liver tumors. Peraino, et al. (1978) also pointed out resemblances between phenobarbital and PCBs. Both substances "enhanced hepatic tumorigenesis" when given after DEN, and both "exerted a protec tive effect against hepatic tumorigenesis" when administered concurrently with AAF or DEN. The comparison between PCBs and phenobarbital is extended by the observation of Berry, et al. (1978, 1979) that PCBs did not promote skin tumors in mice pretreated with DMBA. Peraino, et al. (1978) refer to a study4 in which skin tumorigenesis was not enhanced in mice fed phenobarbital after skin painting with DMBA. Lichti, et al. (1978) described the induction of ornithine decarboxylase (ODC) in mouse epidermal cell cultures by TPA. Aroclor 1254, apparently at much higher concentrations than TPA, induced a low but reproducible stimulation of ODC in the same system. They also noted a report5 that a high intraperitoneal dose of Aroclor 1254 induced ODC in the liver of rats. After commenting that PCBs "warrant further investigation into their - 22 - SCM 019674 00035r HARTOLDMON0023609 possible action as tumor promoters," they added that "These compounds are being tested on carcinogen-initiated mouse skin (T.J. Slaga, personal communication)". The series of publications by Berry, et al. (1978, 1979) and DiGiovanni. et al. (1977) show that PCBs are not promoters on mouse skin. Authors of some of the mutagenicity studies to be discussed (Danz and Urban, 1980; Norback, et al. , 1981; and Stadnicki, et al., 1979) have suggested that PCBs may act as promoters of carcinogenicity. In general, those comments appear to have been offered as hypotheses to aid in understanding the observations which had been made. This also appears to be the case for the NCI study on Aroclor 1254 which was reviewed by the Data Evaluation/Risk Assessment Subgroup of the Clearinghouse on Environmental Carcinogens. That group, charged with the responsibility of providing a peer review of NCI bioassay reports on chemicals studied for carcinogenicity, made and accepted a motion to add the following to the report summary: "Based on the liver proliferative lesions in the treated rats and published reports, it is suggested that Aroclor 1254 may be a tumor promoter" (NCI, 1978). With respect to tumor promotion, Weisburger and Williams (1980) state that "certain inducers of liver metabolic enzyme systems, such as phenobarbital, DDT and BHT, when administered after minimal doses of primary hepatocarcinogens exerted a powerful promoting effect". As mentioned earlier, PCBs also induce liver metabolic enzyme systems. However, since the carcinogen alone produced tumors in animals in the co-carcinogenesis studies summarized in Table 3, it appears that the doses were greater than minimal. In addition, even though the co-carcinogenesis studies were of ' 23 " SCM 019675 onQ"53 HARTOLDMON0023610 shorter duracian, severalused much higher dietary levels of PCBs than those fed to rats for 2 years. The latter studies are the ones which gave rise to questions of carcinogenicity. Thus, while the evidence indicates that PC3s are not carcinogenic, their reported effects in rodent livers following prolonged exposure in conjunction with an initiator may be promotion or inhibition. ' When administered to animals together with a biological agent, PCBs show a promoting or inhibitory effect similar to that seen with, chemical agents. Pretreatment with PCBs inhibited the growth of Walker 256 carcinosarcomas in rats and increased their survival time (Kerkvliet and Kimmeldorf, 1977a,b), enhanced the growth of MSB tumor in mice (Kerkvliet and Roller, 1978) and neither promoted.nor induced the oncogenesis of Moloney leukemia virus in mice (Roller, 1977). ` Mutagenicity Studies Mutagenicity testing of PCBs has ranged from bacterial systems to intact animal studies. A series of studies with eleven bacterial test strains (Heddle and Bruce, 1977; Hsia, et al. 1978; McMahon, et al. 1979; Odashima, 1976; Probst, et al. , 1981; Sugimura, et al. , 1976; and Wyndham, et al. , 1976) are summarized in Table 4. Hsia, et al. (1973) used both phenobarbital and Aroclor 1254 to induce liver enzymes in rats for preparation of S-9 activation systems. They also tested 4-hydroxy2,2',5,5l-tetrachlorobiphenyl and 2,2*,5,5'-tetrachlorobiphenyl-3,4-oxide, a known and a presumed metabolite of 2,2' ,5,5'-tetrachlorobiphenyl, respectively. All three materials gave negative responses with each activation system. Odashima (1976) presented data in tabular form from a series of screening tests. In addition to the results shown in Table 9, " 24 - . $CH 019676 0003G(i HARTOLDMON0023611 bacterial strains WP2, TA100, TA98, H-17, M-45, W3110, and TA1973 are shown in groups in their Table 5 with a text notation that not all chemicals were tested with each strain. For Kanechlor 300 and 500, such of the preceding strains as were tested showed a negative response. The group consisting of H-17 & M-45, WP2 try' (her1- & her') shows a plus sign for Kanechlor 300. Presumably, one or more of those bacterial strains gave a positive response. Kanechlor 500 was listed as giving a negative response with the latter group. With the exception of Wyndham, et al., (1976), all of the studies were negative with and without the addition of a microsomal enzyme activation system. It is somewhat difficult to reconcile the text and the graphs in the publication by the latter authors. They stated that their "results clearly showed that as the degree of chlorination decreased themutagenicity increased, a concentration of 100 pg of 4-chlorobiphenyl in the test medium gave over 2000 revertant colonies per plate. The higher chlorinated biphenyls show very little activity as mutagens". While Figure 3 in their article shows their marked increase in revertants for 4-chlorobiphenyl, the same figure also shows results for Aroclor 1221, which averages 1.15 chlorine units per molecule. The mutagenicity of Aroclor 1221 is not discussed in the text, but Figure 3 shows that at a concentration of 100 pg per plate, it produced about one-tenth of the revertant colonies that 4-chlorobiphenyl did. Thus, a 15% increase in chlorine content produced a 10-fold decrease in the reported mutagenic activity. In light of that sharp reduction of mutagenic activity with a slight chlorine increase, it is difficult to understand the statement in their text that Aroclor 1254 "was only weakly mutagenic." Results from Aroclor 1254 (average of 4.96 chlorine per molecule) are not shown in their ' 25 ' . SCH 019677 000361 HARTOLDMON0023612 Table 3, but 2,2' ,5, 5!-tetrachlorobiphenyl (average of 4 chlorine per molecule) was presented. At 100 pg per plate, 2,2',5,5'-tetr3chlorobiphenyl was virtually devoid of mutagenic activity. Overall, primarily because McMahon, et al. (1979) reported that 4-chlorobiphenyl was not mutagenic, it can only be concluded that the findings of Wyndham, et al. (1976) are aberrant. A similar conclusion that the findings reported by Wyndham, et al. (1976) are unfounded because attempts to repeat them have been unsuccessful was reached by the State of California (Anon. 1981). In a variant of this test, Stott and Sinnhuber (1978) used Aroclor 1221, 1242, 1254, and 1260 to induce the mixed function oxidase system of the liver in trout. The subraitochondrial fraction of those livers was used to activate the metabolism of AFBi which was assayed for mutagenicity using strain TA 1538 of S. typhimurium. The mutagenic response was decreased compared to the concurrent control using untreated trout liver. A general pattern of decreasing response with increasing degree of chlorination was observed, except for Aroclor 1260. Induction of mutagen detoxifying enzyme systems was suggested as a possible explanation for the apparent conflict between the reported induction of trout mixed function oxidases and the decrease in mutagenic activity. These results are consistent with those of Hendricks, et al. (1977) discussed earlier who reported that Aroclor 1254 reduced hepatic tumors in trout fed AFBX. On the basis of slower sedimentation rates in alkaline sucrose gradients, Stadnicki, et al. (1979) reported that 2,2`,5,5'-tetrachlorobiphenyl (TCB), a mixture of the 3-hydroxy and 4-hydroxy derivatives of TCB and the 3,4-epoxide of TCB induced single strand breaks in DMA of L-929 cells. - 26 SCH 019678 00Q36* HARTOLDMON0023613 At 100 pg/ml. each of che three materials caused all of the DMA co come out in fractions at Che cop of che gradient. The epoxide caused some breakage down Co 1 pg/ml. The mixture of hydroxy derivatives caused significant breakage at 20 pg/ml and only slight breakage at 1 and 10 pg/ml. TC3, che lease pocent, caused lesser breakage at 20 pg/ml and was without effect at 1 and 10 pg/ml. Nilsson and Ramel (1974) conducted genetic tests on adults and larvae of Drosophila melanogaster fed Clophen 30 and Clophen 50. These PCB mLxtures were wichout effect on the loss of sex chromosomes used Co measure chromosome breaking action and nondisjunction of the sex chromosomes. Tazima (1980) has described a specific locus test using the silkworm (Bombvx mori). Neither Kanechlor 300 nor Kanechlor 500 showed mutagenic activity in that system. ' Hoopingarner, et al, (1972) induced mitosis in cultured human lymphocytes with phytohemagglutinin and treated them with 100 ppm of Aroclor 1254. There was no effect on the mitotic index, satellite association, chromatid gaps or chromatid breaks in comparison to control human lymphocytes. Odashima (1976) also studied the incidence of chromosomal aberrations. Their in vitro test used Yoshida ascites sarcoma cells cultured for 6 to 72 hours in the presence of PCBs at concentrations producing a minimal or 50% growth inhibition. Kanechlor 300 gave a positive response and Kanechlor 500 a negative one. For an in vivo system, they examined bone marrow cells 6 to 48 hours after adult or newborn animals were given an approximately lethal dose of PCBs. This test gave the opposite result; Kanechlor 500 was positive and Kanechlor 300 was negative. They noted " 27 " SCH 019679 HARTOLDMON0023614 that chromosomal aberrations frequently occurred in controls and that chemicals were called positive when they induced more than twice the aberrations In controLs. In "a Syrian hamster cell transformation system (Pienta, 1980), Aroclor 1254 was one of several chemicals tested double-blind. It gave a negative result. Mouse embryo fibroblasts alsohave been exposed to different PCBs. Nesnow, et al. (1981) studied the cocarcinogenic action of agents which increase microsomal mixed-function oxidase activity in the C3H10Tl-sCL3 transformation assay. After a 48-hour pretreatment with Aroclor 1254, cells were then treated with benzo(a)pyrene [B(a)P] and the agent for an additional 24 hours. Aroclor 1254 did not increase B(a)P-mediated transformation and no Type II or Type III foci were observed. Norback, et al. (1979, 1980, 1981) exposed CSHIOT^ cells continuously for 6 weeks to 10 pg Aroclor 1254 per ml of medium. Treated cells developed Type III foci. Cells exposed to the same concentration of Aroclor 1254 for 24 hours or to 1 pg of Aroclor 1254 per ml of medium did not develop Type III foci. Continuous exposure of cells to Aroclor 1260 and 2,4,5,2',4',5'-hexachlorobiphenyl also caused formation of Type III foci. A clone from a focus transformed by Aroclor 1254 induced sar coma formation when inoculated in irradiated mice. Foci from Aroclor 1260 and 2,4,5,2',4',5'-hexachlorobiphenyl had not been characterized further. Since transformation occurred only after continuous exposure, Norback, et al. (1981) suggested that the effects of PCBs in culture include promotion. When fed to rats, however, 2,4,5,2*,4*,5'-hexachlorobiphenyl produced neoplastic liver nodules, not hepatocarcinomas (Weltman and Norback, 1979). - 28 SCM 019680 0003b* HARTOLDMON0023615 Wong, et al. (1979) claimed that 4-chlorobiphenyl induced an increase in unscheduled DN'A synthesis in Chinese hamster ovary cell cultures in the presence of hydroxyurea, a chemical agent which suppresses normal replicacive DN'A synthesis. Few details were presented regarding the validation and reliability of their test system. By contrast, Aroclor 1254 gave a negative response in an unscheduled DNA synthesis in primary cultures of adult rat hepatocytes (Probst, et al.. 1981). The latter authors presented results of an extended series of compounds tested in their system. Danz and Urban (1980) have proposed using the mitogenic response of the rat adrenal cortex after dosing the animals with a test substance as a short-term test for evaluating the promoting action of chemical compounds. Clophen A60, Delor 103s, Delor 106s, and Phenoclor DP6 all gave positive responses in their test system. At 3-6 days of incubation, embryos from ring doves (Streptopelia risoria) fed a control diet or one containing 10 ppm of Aroclor 1254 were examined for cytogenetic changes (Peakall, et al., 1972). The relative frequencies of chromosome aberrations in the . 8 largest chromosome pairs in metaphase cells of allantoic sac and limb bud origin were scored. The 6 control embryos had a mean aberration rate of 0.8% (0-2.0). The aberration rate in 17 embryos from Aroclor 1254 treated birds was 1.8% (0-9.4). In the latter group, there was one chromosome rearrangement, 13 embryos with aberration rates exceeding the mean control rate, and 4 embryos which exeeded the highest control rate. Aroclor 1242 was injected into fertile White Leghorn eggs to give estimated final concentrations of 10 or 20 ppm (Blazak and Marcun, 1975). After 4 or 5 days of incubation, eggs were - 29 SCM 019681 000365 HARTOLDMON0023616 injected with coicemide, incubated an additional 45-60 minutes, and embryos were harvested for examination. There was a high degree of early embryonic death, and a few live embryos showed drastically retarded development without malformation. The first five pairs of chromosomes, constituting over 50% of the chromatin material per cell, were examined for detection of clastogenesis. There were no chromosomal aberrations. Heddle and Bruce (1977) injected mice with Aroclor 1254 for five consecu tive days and then examined bone marrow preparations for chromosomal breakage and sperm ceE preparations for sperm with abnormally shaped heads. While nonspecific factors can induce sperm abnormalities, the authors believe the latter also can result from point mutations or small deletions. Aroclor 1254 was judged to be neither carcinogenic or mutagenic. ' Dikshith, et al. (1975) gave male rats oral dosages of 50 mg/kg of Aroclor 1254 on each of 7 consecutive days. Animals were killed at intervals over the next 3 days. Those scheduled for cytogenetic analysis were injected with colchicine 2 hours before killing, after which time the seminiferous tubules were prepared for such study. At autopsy of the other animals, testis, epididymis, and liver weight were recorded and sections were prepared for microscopic study and histochemical determi nations. Livers were markedly enlarged and showed a significant increase in weight compared to controls. There were no differences in body weight or weight and appearance of testis, epididymis, or vas deferens between control and treated rats. Aroclor 1254 treated rats showed a few meta phase figures with abnormal chromosomes whch appeared to be sporadic and not specific to any one type. Histological examination showed testis - 30 JU SCH 019682 00Q36o HARTOLDMON0023617 and epididymis from Aroclor L2S4 treated rats were comparable to controls although interstitial cells were increased in Aroclor 1254 dosed rats. These cells showed an increase in acid phosphatase activity. The authors concluded that they "found no evidence to suggest that Aroclor 1254 causes significant chromosome damage or histopathological changes in the rat testis." ' Similar studies were conducted by Green, et al. (1973, 1975a) who gave male rats single oral dosages of 5000 mg/kg, 2500 mg/kg or 1250 mg/kg or four successive daily dosages of 500 mg/kg of Aroclor 1242. Other rats received five consecutive daily dosages of 300 mg/kg, 150 mg/kg or 75 mg/kg of Aroclor 1254. Animals were injected with colcemide 3 or 4 hours before sacrifice which occurred about 24 hours after the single dose ` or the series of doses. There were some body weight losses and some ' deaths occurred. Bone marrow from rats dosed with both PCB mixtures and spermatogonial preparations from Aroclor 1242 treated rats were prepared for cytogenetic study. Repeated dosages of 150 mg/kg and 300 mg/kg of Aroclor 1254 produced decreases in the number of mitoses in bone marrow cells. Repeated dosages of 75 mg/kg of Aroclor 1254 and all dosages of Aroclor 1242 were without effect. Neither PCB mixture at any dosage produced a significant number of chromosomal abnormalities in bone marrow cells. At the lower dosages, Aroclor 1242 did not affect mitoses of spermatogonial cells, but 5000 mg/kg or 4 dosages of 500 mg/kg significantly reduced the rate of cell division. No dosage of Aroclor 1242 produced cytogenetic abnormalities in spermatogonial cells. The authors concluded from their studies that Aroclor 1242 and Aroclor 1254 did not possess mutagenic potential. Garthoff, et al. (1977) fed male rats diets containing 5, 50, or 500 ppm of Aroclor 1254 for 5 weeks, after which they " 31 " SCM 019683 000367 HARTOLDMON0023618 examined bone marrow and testis samples. There was no significant difference between test and control animals with respect to the incidence of chromosomal abnormalities and the number of cells in mitosis from bone marrow and spermatogonial cells. In a dominant lethal study (Keplinger, et al.. 1972 and Calandra, 1976), albino mice were given single intraperitoneal dosages of 500 or 1000 mg/kg of Aroclor 1242, Aroclor 1254, or Aroclor 1260 and mated on successive weeks to virgin females. There was no evidence of mutagenic effects. Although details on their studies are limited, their results are in general agreement with those from another dominant lethal study in rats conducted by Green, et al. (1975b) with Aroclor 1242 and Aroclor 1254. The former was administered orally at a single dosage of 625, 1250, or 2500 mg/kg or in five daily dosages of 125 or 250 mg/kg. Aroclor 1254 was given orally in five daily dosages of 75, 150, or 300 mg/kg. After dosing, they were mated with untreated females for 10-11 weeks. Another group of rats was given 150 mg/kg of Aroclor 1254 for five successive days and starved overnight before admittance to females. Other male rats were offered diets containing 25 or 100 ppm of Aroclor 1254 for 70 days, then mated with untreated females for one week. TEM (triethylenemelamine) was used as a positive control. There was a body weight loss and some deaths occurred in a few groups. Neither the oral dosing nor the dietary feeding of Aroclor 1242 or Aroclor 1254 had any effect on the number of implantations or the number of dead implantations per pregnant female while TEM produced significant postimplantation losses in weeks 2, 3, and 4. These authors noted that the reduction in the number of dividing spermatogonial cells reported earlier for Aroclor 1242 (Green, et al., 1973, 1975a) did not impair the reproductive performance of male animals. - 32 SCH 019684 (V^ "6^ HARTOLDMON0023619 Review of the various mucagenicity tests and their results prompts three comments. Chlorinated hydrocarbons do not generally give positive results in Salmonella strains. Therefore, the mutagenic responses with TA1538 (Wyndham, et al. 1976) are either aberrant or else the response is, indeed, Limited to essentially monochlorobiphenyl. The degree of correlation between in vitro mutagenicity tests and carcinogenicity depends upon the initial classification of the test materials. Pienta (1980) classifies Aroclor 1254 as a non-carcinogen. Rinkus and Legator (1980) regard Aroclor 1254 and Kanechlor 500 as having known or suspected carcinogenic activity. Odashima (1976) and Sugimura, et al. (1976, 1977) consider Kanechlor 500 to be carcinogenic and Kanechlor 300 to be non-carcinogenic. Apart from the difficulties they present for correlation, these different opinions about the carcinogenicity of PCBs are a reflection of what data these individuals considered and how they evaluated those data. Finally, in light of the large number of tests conducted to evaluate various mutagenic parameters, it is not surprising that an occasional suspicious or positive finding resulted, simply on a statistical basis. Those occurred in in vitro systems. In vivo tests gave negative results and provide a basis for concluding Chat ambient levels of PCBs do not present a mutagenic risk. Epidemiology' Studies With respect to epidemiologic studies, it should be noted that there are frequent references in the literature to an epidemiologic study of workers exposed to Aroclor 1254 (Bahn, et al. , 1976, 1977). Those articles are cited in two recent reviews. One states "The epidemiological data provide suggestive evidence of a relationship between exposure to polychlorinated biphenyls and the development of malignant melanoma. ...for practical - 33 SCM 019685 00Q36. HARTOLDMON0023620 purposes, polychlorinated biphenyls should be regarded if they were carcinogenic to humans" (IARC, 1978). The ocher states "No conclusive evidence has chus far been reported which demonstrates that occupational exposure to PCBs has caused an increased incidence of cancer" (Kimbrough, 1930). The latter author then proceeds to discuss the study by Bahn, et al. (1976). Among the references for these 2 reviews are NIOSH (1978) which has the following comment on the Bahn study, "PCB exposure histories were based on recollections of two company employees. Exposures to other chemicals could not be ascertained.------ To correct these deficiencies in the preliminary study, a more intensive investigation is being conducted (B.N. Kightlinger, written communication, November 1976). A substantial change has occurred in the cohort since release of the preliminary report by Bahn and her coworkers, and it seems likely . that the findings on this new cohort will differ significantly from those of the preliminary study. The final report is not yet available." Recently, Gaffey (1981) reviewed and evaluated existing reports concern ing health effects and exposure to PCBs. The reports he discussed dealt with diverse potential health effects. With respect to liver effects, he concluded that "Alterations of liver function and fat metabolism associated with PCB exposure have been observed in several studies, but are characterized by investigators as mild and of no clinical significance." He also concluded that "Mortality studies concerned primarily with cancer present problems of interpretation due to the small sample size of some of the studies, and to the confounding effect of other exposures. However, they do exhibit a pattern, which is that none of the studies agree on the cancer sites at which an excess mortality was found, and the excesses that ` 34 " SCM 019686 OHO^'O HARTOLDMON0023621 were found are in general not statistically significant. One must conclude that the findings of the mortality studies reflect a sporadic pattern of excess mortality at different sites which is not consistent with a carcinogenic effect of PCBs. In addition, where an examination of duration and latency of exposure was possible, no association with these variables was found [32]7 . ' "Taken as a whole, the epidemiologic studies find that high occupational exposures to PCBs may cause dermatitis of various kinds, but that there are no other clinically observable effects, including the occurrence of cancer." Summary Some remarks by Cole and Merletti (1980), although written in a more ' general vein, appear to be particularly suited for ending a discussion on the potential carcinogenicicy of PCBs. "In view of the difficulty of deciding whether a "suspect" carcinogen is in fact a weak carcinogen or in fact harmless, we point out one aspect of this scientific judgement which, though well known, is often lost sight of; namely, that in prin ciple, it is more likely that a non-carcinogen will appear to be a weak carcinogen than the reverse. This asymmetry should be fully appreciated by legislators and regulators. It is summed-up succinctly, if not very accurately, in the oft-heard phrase "you can prove a positive but not a negative". This is clearly illustrated in the case of PCBs. Review of the results of a large number and wide range of chronic feeding and metabolism studies in animals and of mutagenicity studies in various systems has failed to establish that PCBs are carcinogenic. However, - 35 - SCM 019687 (JHM.rVl HARTOLDMON0023622 presently available rodent data raise some suspicions and lead to disputes about the carcinogenicity of PCBs. Attempts to compare and evaluate results of rodent studies reported in the literature are complicated by different uses of similar terminology and variations in the criteria employed for diagnosing hepatic tumors in rodents. Co-carcinogenesis studies have reported both promotion and inhibition of tumors in rodent livers following prolonged administration of PCBs in conjunction with an initiator. Thus, it is not likely that animal studies and other laboratory procedures will lead to a universal consensus on the carci nogenicity or non-carcinogenicity of PCBs. While epidemiology studies of humans exposed to PCBs present problems of interpretation due to their small sample size and the confounding effect of other exposures, they present the best available evidence for assessing the carcinogenic potential of PCBs to humans. Epidemiology studies, including recent studies involving large cohorts with lengthy exposures to PCBs, demonstrate a pattern that is not consistent with a carcinogenic effect of PCBs. - 36 - SCM 019688 HARTOLDMON0023623 Footnotes 1. Farber, E. (1973). Hyperplastic liver nodules. In:Methods in Cancer Research, Vol. 7, H. Busch, ed. Academic Press, N.Y. pp. 345-375. 2. See reference: Squire, R.A. and Levitt, M.H. (1975). 3. See reference: NCI (1978). 4. Grube, D.D., Peraino, C., and Fry, R.J.M. (1975): The effects of dietary phenobarbital on the induction of skin tumors in hairless mice with 7,12- dimethylbenz(a)anthracene. J. Invest. Dermatol., 64. 258-262. 5. Costa, M., Costa, E.R., Manen, C.A., Sipes, I.G., and Russell, D.H. (1976): Adenosine cyclic 3',5'-monophosphate-dependent protein kinase and ornithine decarboxylase involvement in the induction of cytochrome P-450 and hepatic hypertrophy. Mol. Pharmacol., 12, 871-878. 6. Delor is a tradename for PC3s made by Chemko in Czechoslavakia. 7. Brown, D.P. and Jones, M. (1981). Mortality and industrial hygiene study of workers exposed to polychlorinated biphenyls. Arch. Environ. Health 36, 120-129. SCM 019689 000373 HARTOLDMON0023624 References L. Anon. (L98L). The Toxicology of PCB's. An overview wich emphasis on human health effects and occupational exposures. Hazard Evaluation System. Epidemiological Studies Section. State of California. Berkeley. 2. Bahn. A.K., Grover, P., Rosenwaike, I., O'Leary, K., Stellman, J. ' (1977) PC3? and melanoma. Hew Engl. J. Med. 296, 108. 3. Bahn, A.K., Rosenwaike. I., Herrmann, H., Grover, P., Stellman, J. and O'Leary, K. (1976). Melanoma after exposure to PCB's. Hew Engl. J. Med. 295, 450. 4. Berry, D.L., DiGiovanni, J., Juchau, M.R., Bracken, W.M., Gleason, G.L., and Slaga, T.J. (1978). Lack of tumor-promoting ability of certain environmental chemicals in a two-step mouse skin tumorigenesis assay. Res. Commun. Chem. 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Devenish, J. , Safe, S. (1976). The in vitro metabolism, macromolecular binding and bacterial mutagenicity of 4-chlorobiphenyl, a model PCB substrate. Res. Comm. Chem. Pathol. Pharmacol. 15, 563-570. 88. Yoshimura, H., Yoshihara, S., Ozawa, N. and Miki, M. (1979). Possible correlation between induction modes of hepatic enzymes by PCBs and their toxicity in rats. Ann. N.Y. Acad. Sci. 320, 179-192. SCM 0L9697 0^0 rri HARTOLDMON0023632 TABLE 1 PCBs: Tal>ulalion of Liver Nodules Reported in Mice1 Approx. wt. X Cl2 Product Dosing Strain Duration PCB in diet (ppm) No. Animals Nodular Hyperplasia Hepatoma Hepato cellular Carcinoma deference 52-54 Aroclor 1254 BALB/cJ 6 mos3 11 mos Kanechlor 500 dd 32 weeks Kanechlor 500 dd 32 weeks Kanechlor 500 dd7 32 weeks 0 300 0 300 0 100 250 500 0 100 250 500 0 100 250 500 34 24 24 22 6 12 12 12 20 18 20 17 12 10 20 17 - 0 0 0 7 9 4 0 1 0 9< (-)5 '- - Kimbrough - +,Linder (1974) - 0 Nagasaki, et a 1. 0 (1972) 0 1 to, et al. (1973a.b) 5s 0 Nagasaki, et a 1. 0 (1974, 1975) 0 76 0 Nagasaki, et a 1. 0 (1974, 1975) 0 0 48 t/i n X o 3o O *.) o 'a CD Kanechlor 400 dd Kanechlor 400 dd Kanechlor 400 dd7 32 weeks 32 weeks 32 weeks 0 100 250 500 0 100 250 500 0 100 250 500 6 12 12 12 0 17 19 20 12 20 20 17 0 0 0 0 - - - _ *- . - - 0 Nagasaki, et al. 0 (1972) 0 Ito, et al. (1973a.b) 0 0 Nagasaki, et al. 0 (1974, 1975 S ' 0 0 0 Nagasaki , <-L a 1 . 0 (1974, 1975) " 0 0 *, HARTOLDMON0023633 TABLE 1-continued Approx. wt. X Cl2 Product Strain PCBs : Tabulation of Liver Nodules Reported in Hice1 Dosing Duration PCD in diet (ppm) No. Auima1s Nodular Hyperplasia Hepatoma llepatocellular Carcinoma i Refereure 40-42 Kanechlor 300 dd 32 weeks 0 100 250 500 6 12 12 12 o' 0 0 0 . 0 Nagasaki, e i a 1 . - 0 (19/2) - 0 [to, eL al . ( 1973a,h) -0 t Kanechlor 300 dd 32 weeks 0 100 250 500 20 19 19 20 - - - - 0 Nagasaki, eL a 1 . - 0 (1974, 1975) -0 "0 Kanechlor 300 dd7 32 weeks 0 100 250 5110 12 19 20 20 - - - 0 Nagasaki, el a 1 . - 0 (1974, 197 5) -0 0 1 Hales, except as noted. 2 From Brinkman and deKok (1980) * Held an additional 5 months before sacrifice. 4 Ho. of animals. One had 2 hepatomas for a tumor total of 10. 8 Apparently same data reported as hepatoma and hepatocellular carcinoma. See references and text. a Apparently same idata reported as nodular hyperplasia and hepacellular carcinoma or Lumor. See references ami text. 7 Females. IoA z o o o Q-- *0 0" 00 CJ HARTOLDMON0023634 ! TAUI_.E_2 FCHs: T*iluidi iua of I.iver Nmlulcu Hcporteil in HuLa Approx, wt . I Cl 1 Product. Strain Sen PCD Doling in diet No. Dural ion _!ppiu)___ Aiiiiu 1 s Arii'iiouijioua Nodules Nodular iiyjicrj11 us i a 60 Aroclor 1260 Site man V 21 nos.^ 0 in 100 184 - _ - Aroclor (21*0 Churle* M, 24 aioa. Hi vcr K 212\414\S,5` - - Hdvcbloro6i phciiy la - 24 noi. 0 1 10 100 100 21 26 25 25 - - - 1 0 7 6 - N(`0|i 1 usl i c Nodu1es Adenoma Ilepul Iiiuj Ilepaloic 11 o 1 j r Oj 1 11 lliMU.I Clio- I.IIRIOlli`p.1 ( tiou Hr 1 i-i'piiir 0 __ 1 144 " - 26 _ K i ml*i ougit, - cl_.ul. (l`J/5) - _ 0 _ 0 i.t:v titsk.is * - - (1 - 0 ( 1 `J 8 1 ) - - 1 - (1 - -7- 4 < -. _ We 11 uiii u 6 Nurl.jck (I'J/'JI 52-54 Aroclor 1264 Clurle* N. 24 nos. 0 2'J 1 0. 0 Ktver F 1 30 - 0 ' - - 0 - 0 (IU8I) 10 26 - 3 - -0- 0 100 26 - 14 - -4- 2 Aroclor 1254 Fic|ir N 105 weak* 0 24 . . . 0 . 0 m NCI (10781 144 26 24 - - - 00 . SO 24 - - - 0 - 1 _ 100 24 - - 1-2 - Aroclor 1254 Fischer F 105 week* 0 21 - _ 0_ _ . NCI ( I'J/H) 344 26 24 - - - 0-- so 22 - - - 1-. _ 100 24 - - 2- - - Aroclor 1264 l.oCa 1 F 28 uui.6 0 u - - - I) - _ _ i 111.* II , 200 6 " ~ " i4 " - - e 1 .*1 . ( I'J/ll) Kjitcklilur UiiiUr n 28-52 wk 0 )H - 0 - -._ 600 IlMl 2* - 1 * -- - - ( 1`1/C) MU) )< - 6 - --- . 100)) 1) 6 - --- - .. ........... . . _. HARTOLDMON0023635 SCM 0 1 9 7 0 0 I A|>|iroM. Ul. 1 Cl 1 produil 46 Kailcclilor 400 Kaiieiiilor TAIII.K 2-cunl iiiU4*I PCjji: Taiuijat imi u| Liver Nmlnlus Hi^iurinl iii Hal a pc:h Dosing in ill el No. Alrmuit.i! mis Noiiu 1 a r Si rain Sl-x Dural ion (|l|Hll) Aii im.11 * Noitu 1 rs lly|u*i |> 1 as l a Doiiryu ti 22-62 wks 26.5- 5 t) . till 10 t) . - K 50 - ID (. - WtkLur ti 26-52 wkx 0 1D0 500 lOUO IH 16 6 10 . 0 2 0 a N<*o|il.isi ii: Nmlnlrs . - - . . AJriioma - . llrjiat oiua - -' llr|l.ilbirl iii la r Ca 11 l iiouiu - . . - Clio1 aiiK i u 11 4 ' |4. 1 Will. _ . - _ _ _ Hi* I * i i* luV Kiiiuua i ll.ilta d'j/ n 11 , ri ,i i. (i ')M ' 40-42 Aroclor 1242 (Mur 1 cm Hi 24 MOS. II i ve r K 0 1 10 100 Kxnedilor Wiklar H 26-52 wkx 0 100 *00 1000 2,2' ,5,5'- iclrachlorobi|i|ienyl - - 24 aios. 100 21 22 20 IV IB 22 1!) 15 - - 1**4 - - -' i i i B 0 1 0 0 - . 11 IJ l.*v i nokat> - 0 - (1 ( I'JBI ) - - 11 - II - - a- - 1 _ __ . I lu, 1*1 .1 I . ----- U'J/O' . - --- - .__ W: I 1 uiaii L Hui I.ji k ( O/'M 1 Krou briukuuan ami JeKuk (IDBO). * llrlil aii adl111oiial 2 uionliis Itclofc kacrilirc. * Absirail. Drlii h> liiailcil. 4 I'icfct'iiic rcjoricii. * Llliaaul solution iii ill (iik int; wjlcf. SCM 019701 HARTOLDMON0023636 SiBrJO U U SCM 0 1 9 7 0 2 Product Kaneclilor 400 KjiicdiUr SOU Kdiittlilor SOU KaneClilor 400 Kanechlor S00 Kauechlor 500 Aroclor 1254 Kanechlor 5002 Aroclor 1254 KjiiediUr 500 KdiitfClilur 500 Kdiurt.li lor 500 Kdiicihfur 500 TABEEJ co-cakcino<;khe.sis studies with mu Hade Tmc Treatment1 Mode Species Observalions h!c 1 . Diet Willi Diet WlLll Dicl Did Willi Villi Did Willi Cavage After Did Gjvng After lie I'e re Diet Willi Hi eL With Diet Willi lIlLl llclurc Hint Alin 2a-HcCli If-line ill cr inf: |UI(l|l4it Iliirt (1-UIIC Hurt O-UIIC Did DEN Wilier DEN Water HEN DEN* Water Water AKH, JlieL 2-KAA Diet `J * -Me-IIAH Dili J'-H:~HAU lint 1 ' -fle-IMII Jii.i Jil mouse tlt| amuse ild mouse dil mouse Sprague* Hawley rat Wislsr rat Spragne* Hawley rat Wifctar . rat Ka Inliow t root S|ii ague* Hawley rat SpragueHawley rat S|H dgot:Hawley iat S| agile * llawley i il llcpal ore 11 ii|ar carcinoma witli 2H~hcCb. Node with cowl* mat lull. Comb i na l i nn i nercased liepal uce 1 1 u 1 ar Carcinoma over u*llllC alone. Combi nut Inn praOuced licpalocel lular Carcinoma. Each alone negative. Combination Increased liepalucel lular carcinoma ovor u-UIIC alone. Hepatocellular Carcinoma with DEN. None with combination. Combinat ion increased hepatocellular carcinoma liver HEN uloite. Combination increased lii?|iuloci;l lular carcinoma over HEN alone. Combination decreased liepalord lular carcinoma in oil*firing. Hepatocellular cjrcinomJ with AKMj. None with combination. Ilepatorcl lular carcinoma with 2-KAA. None willi Combinat ion. Ilrpat tite 1 1II1 ar carcinoma willi J1 -tle-HAll. `Nunc with couili 1 Hal i on . ilepa l me 1 1 ii 1 a r ea rc I inuna willi l**He~|iAH. Nuni! willi comb 1 lial 1 mi. I'tMiib j |* .11 l oil int*ie.ised Itep.t 1 >i e | 1 u 1 >i i i.ii> i ii>uvi-i 1` fli,` llftll .iltim'. III b 1 yama , and lTit i b.i, | `1 /4 11 u, el a 1 . rM u.i. Nagasnk 1 , d i{. I`J/A, iy/*i 1 1 ii i 1*1 a 1 . 10/iaJ* NagaSaki, el al. 1 *J 74 , nm Nagasaki, et al. ) `i 7 4 , i*m Mukiura, el al: 1074 Nisiiiaumi, l`J76, |*j,, i. l`res Ion, el a | . 1 *>H I Nisillaumi , iyuu Hendricks, el al. iy/7 Hakiura, e 1 al. |)74 flakl in a , e 1 . 1 . 1*1/4 ' K imm a , el al. I'j/li Kiiiiiii .t, el al i'i/ K MUD 1 .> t t a| J`l/,, HARTOLDMON0023637 H 019703 000.<Sv Product Kaueclilor 500 Kaueclilor 500 Kanechlor 500 Kanoclilor 500 Kancchlor 500 Unspecified Kaiiadtior Aroclor 1254 Aroclor 1254 Aroclor 1254 Aroclor 1254 Ai or 1 ti 1254 A.ot |r 12^4 Mode Diet Old Cavage Ctvage Gavage Diet Skin Shin Sk in Did Dili Did Time With With After l> Ullll After 4 Willi After l With Alter i With Alter lit fore lit: fore ltd uTc lie lure |le|.c Other Treatment1 TAIJI.K 7-cuiil iuuoil SUMMAUY OK <:Q-CAIlc:iNi>GK.NKSIS SHIM IKS WITH Mis Hode Species Observalions Kt: 1 . i'-Me-UAU * DEN Diet Sprague* Oawley rat 2-KAA t UEN IlieL Sprague** Oawley rat I1KH I1MT Water Gavage IIKN Mi'll Water Water DEN Mi'll, l)l)T Voter Voter, Uovoge 2-FAA Petri 1 si lic|ulecioky Diet WisLar rai Wislur rat Wislar rat Pi setter rat 7,12-DMBA Skin C0l Mouse TPA .Skin C|)l mouse ;.i2-nmiA i TI'A Skin Ciil mouse Walker 256 llll fd* liuuoi' Ctrl Ik muscular HSU 4 el U 1 IllL ra~ muscular Mu | unry |rttki'iU|.| Vllllit Ini i |'f | | 1 tiHIM 1 Spiague* llowlry i.it i:miii./i> ll.ilh/t imn:,r Cuiultinul ion decreased hepatoiel lular carcinoma over !i'~Hi:-0AU t 01.N. Cuuibiual ion decreased hepal me 11 ular cat cinema over 2-KAA i 0KN. Combinot ion increased hepalocel lular carcimiMj over DKN * 00T. Climb Inal ion increased hepatord lular carcinoma over DCN SIMI. CombinaLion inrrensotl hepalore 1lular carcinoma over UKH, SPB, OUT. harked Increase lit hyperplastic liver iiudiiltis. tlakiuia, cl a 1 . 19/4 hakiura, cl al. 19/4 * Nikhiicium, 1979a,b Nislii xuuii , 19 79a ,b Nishiauibi, 19 79a , h llu, rl M. Not a promoter of skin tumors. Utile or no skill tumor initiation. Nu <ii 1 ferciice or decrease in skin Luunir induction over 7,l2~bHIIA U TI'A, depending on prel lealmeiit inleival. Tuuiur growth inhibited. `Survival time increased. ' Knham'ed giuwth ol MSII tiinioi . Uerry, l'L ol. I'm, Kerry, i:l al. 19 79 DiGiovaiiiii . el al. 1977 Uerry, el al. 19/9 0 > 6 to valid |, el a). 1977 Kei kv 1 11* l ami K liiuiu: liloi l , 1 9 / / .i ( ii Kcrkvltd and K.illi-i, |9/fl Dili 4|tt| .(llrii 4m ttfirin*:. i u i > 1 holuory 1I'llki'fiil .1 V 1 | us . K..I 1 , 1 *}/ / HARTOLDMON0023638 Product Aruclor 1242 Aroclor 1221 Mode Tine TAItl.E 3-conl limed SUHHAHY OK CU-CAHCINOCKNKSIS STUDIES WITH pfbs Other Treatment 1 Hodc Species Oliscrvji tons Hut Uiel lit: 1 ore belore Moloney 1 eokeiui a virus Moloney leukemia vi rub llit i j* 1|i: 11 oiicj | IIj Ih/c woubt: 1 lit l apei i loiiCti | 11.1 Ih/c mouse Did itol .illerl oncogenesis ol Moloney leukemia vi ims. Did uol allecl oncogenesis ol Moloney leukemia virus. 'Chemical* used: 20-MeCh s 20-aclhylclio)*nlhrene a-BI|C a o-benzena hesachlorlda 0-BtlC = 0-benzcne lieMschlor ide |)EN * dielhy InilrossaUe AFB| * aflauvin Bt 2-FAA a N-2-fluorcnylacciamidc 3*-Me-DAB = 3'-aethy 1-4-diaelliylamlnoagobeiuene DOT = diclilorodiphenyltrirhloroethane SPB = todiua phcnobarbilal 7,12-DDA - 712'-diiuelltylht?iu|<i|4nLhriceue TI'A - l2*()'lelritieLMituyljj|iuiho) - I'J-acrl*t e ^Pregnant daws dosed with Kauechlor 4(11), Oil bpring dosed with OEM. ^Established tissue culture cell line derived froa Moloney sarcoma virus-induced luwors hi Il6 aice. He I . Kol ler, 1 *1 / / Ko1 let, l'J/7 SCH 0 ' " *1 HARTOLDMON0023639 i Tester Strain Product AKDCI.OK 1268 AHOCI.UN USA AHOCLOK USA AH0CI.UK USA Kanerhlor 500* K.iieclilor SOU TABLE A PCIls i_TalmI*I ioii_ol'__Mieroltia j__Mulageiiicily JTcsls S. (yjdi i mu r i mu C3076 D30S2 GA6 TAOS TA100 '"TAtOUU TAI535 TAIS36 "r'AI5'3T TAISJH Neg. Ncg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Ncg. Neg. Neg. Neg. Neg. Neg. Neg. Pox* Neg. E. roll UF2 WP2 nvrA- Hcl . Neg Neg o Neg. Wyndliaiii, el a 1 . t*1 /o Pi oltsl , cl at. I4H I llcddlc and IlfuCe \lH / Wyndliaui, el_al . |4 7t Odasliiuia, 1476 Sugiiuura, cl al. I'J/o 2,2*.5,5-tetraclilorobiphenyl Neg. Wyndliaui, cl d|; |47n 2,2* 5,51-let raclilorobiplieny 1 Neg' Neg* Neg1 Neg' K.ncchlor 300* Neg. Neg. Neg. Neg. K.nechlor 300 Neg. Neg. AHOCLOK 1221 . I'u#' 4*chlorobiphenyl * Ho#' ! *j I! J1" 15 SCM 0 1 9 7 0 5 4-chlorobi jiheuy 1 He*. He*. Neg. Neg. Neg. Neg. 1. All ictlu done with aud without activdlion except us noted. 2. oilier xiraim# icxlcd, xce lexl and relcrcnee. = Nol lex led. Ncg. Ncg - Ncgaltve. - Only levied wiilioul dilivalioii. Neg. Neg. l Neg' s Only Icsled williat'l I val i nil. I'uit1 - l`o> ill ve only with .*l i v.il i mi. ` lliit i* i I .i 1 It. 'i*:w lexl. t Ncg o Neg. Him. till. I'JIU Oilaxluuia, 147b Sugiiunru, el al. |47 Uyinlliatu, el al. 147b Wyudliuiu, cl al. 1471* Neg. Mittalum, cl al. 14 74. ------ HARTOLDMON0023640 ILU X I I i <! HARTOLDMON0023641 decision Analysis. Phil 's -eroy, A. A.. Gaylord, rger, T. 1982. Reducing nduced hospitalization: utilization review. Drug 986. Pharmaceutical inenehc dnig competition ieca of the Drag Price id Patent Term Restota14. Pharm. Med. 1:177- :tolIey, P. D.t Brown, T. usubstitution law coa lition? Ann. Inlem. Med. 1986. Med. Lea. 28:1-2 Ann Rev. Pharmacol. Toxicol. 1987 27.87-1 It HUMAN HEALTH EFFECTS OF POLYCHLORINATED BIPHENYLS (PCBs) AND POLYBROMINATED BIPHENYLS (PBBs)1 Renate D. Kimbrough Center for Environmental Health. Centers for Disease Control, Public Health Service, US Department of Health and Human Services. Atlanta. Georgia 30333 INTRODUCTION Polychlorinated biphenyls (PCBs) are chemical compounds with the empirical formula C|2H|o_,Cln, with n = l--10. They are a mixture of chlorinated biphenyl congeners. Theoretically. 209 such congeners are possible, but at least 20 congeners have never been identified in commercial products. In addition. PCBs may contain polychlorinated dibenzofurans and chlorinated quaterphenyls as impurities. PCBs were discovered before the turn of the century, and the useful industrial properties of mixtures obtained by chlorina tion of biphenyl were recognized early. In 1966 the discovery of PCBs in environmental samples (l) spurred renewed interest in the analysis and toxic ity of these compounds. In recent years many industrial nations have taken steps to control the flow of PCBs into the environment. PCBs and PCB-containing formulations are restricted (an exception is sometimes made for mono- and dichloro-PCB) for most uses, except for categories such as closed-system electrical equipment and hydraulic fluids in mining equipment. Commercial production of PCBs began in the United States in the late 1920s. In 1971, Monsanto Chemical Company voluntarily stopped openended uses of PCBs, and subsequently only the lower chlorinated biphenyls 'The US Government has the right to retain a nonexclusive, royalty-free license in ind to any copyright covering this paper. 87 ,, DEPOSITION * EXHIBIT I 1 S iO/( ooorro HARTOLDMON0023642 88 KIMBROUGH HUMAN HEALTH EFFECTS OF PCBs & PBBs 89 were produced (Aroclor 1242 and 1016). In 1977 ihe company ceased produc tion enlirely (2). Many PCHs manufactured in Ihe past (3) are still in use in old transformers, but even this use is decreasing. The estimated cumulative production and consumption of PCBs in Ihe United Slates in the period 1930-1975 (in millions of pounds) was as follows: total production, 1400; imports. 3; domestic sales. 1253; exports, 150. PCBs are inert chemicals that are fairly resistant to degradation. Because of their stability and lipophilicily, they have accumulated in the environment and in organisms. They have been identified in indoor air (4) at concentrations of into feed for livestock. The flame retardant was called Fircmaster and Ihe magnesium oxide. Nulrimaster. In 1973 some bags of Firemaster were accidentally sold as Nutrimaster and mixed into animal feed. This resulted in widespread contamination in the slate of Michigan (14). Since 1974 PBBs have not been produced in the United Stales, At the time of Ihe exposure little was known about Ihe toxic effects of PBBs. Ten years after the Michigan residents were exposed, no clinical illness has been causally linked to PBB exposure in this group, although chloracne was apparently noted in some workers who manufactured PBB. 0.1 /ig/m1 (5). in fish (6, 7) and other food products, and in sediments from lakes and rivers (8. 9). PCBs have also been identified at varying con HUMAN EXPOSURE centrations in soil (0.01 mg/kg-100 mg/kg) (10). They do not occur naturally. Thus, their presence in Ihe environment is linked with human activities, and Because PCBs are ubiquitous and very persistent in Ihe environment, humans concentrations arc higher in urban and heavily industrialized areas than in have been and will continue to be exposed to them, particularly in in rural and remote areas. However, trace amounts arc also found in remote I dustrialized countries. PCBs may be inhaled in small amounts through ihe air areas, since Ihe air may transport such chemicals over large distances. I or ingested through food. In Ihe United Stales today, people arc primarily Although PCBs are no longer used commercially in the United Stales exposed to PCBs by consuming fish from contaminated waters (9). In the because of their persistence, they are still present in our environment. A past, some farm families were exposed to PCBs from dairy products; these number of transformers and capacitors that contain PCBs. however, are still PCBs originated from coating material used in the inside of silos (15). In in use. Results of laboratory experiments showed that pyrolysis of PCBs at addition, workers who repair transformers and workers who handle toxic temperatures of 200-600C could result in the formation of significant wastes may also be exposed (16). amounts of the more toxic polychlorinated dibenzofurans (PCDFs) (II). The PCB products that were manufactured by Monsanto in the United In February 1981, an electrical fire occurred in a New York State office States had the trade name "Aroclor." The particular kind of Aroclor is building in Binghamton. N.Y. The fire, which originated in a switch gear in identified by a four-digit number. The first two digits refer to the 12 carbon the basement, caused the bushings to crack on a nearby transformer. About atoms, and the second two refer to the percent, by weight, of chlorine in the 180 gallons of PCB dielectric fluid Pyralon (65% Aroclor 1254, 35% chlorin mixture. Thus. Aroclor 1254 contains about 54% chlorine, and Aroclor 1260, ated benzenes and trace additives) were lost. A fine layer of oily soot covered about 60% chlorine. many of Ihe internal surfaces of the 18 floors of the building. Analysis of a The composition of this mixture of chemicals, with different properties, soot sample showed that it contained various isomers of chlorinated di changes once it gels into the environment and into organisms. Some com benzofurans, 2.3,7.8-teirachlorodibenzodioxin, other chlorinated di- ponents of the mixture are more easily degraded in the environment than benzodioxins, and chlorinated biphenylenes. Some of these chemicals are i others. As a result the PCBs identified in Ihe environment resemble Aroclor much more toxic than Ihe PCBs. Because of these findings Ihe Binghamton I 1254 but are not identical to it. Similarly, the PCB mixtures found in humans stale office building was closed, and workers wearing respirators and pro ' usually resemble Aroclor 1254 if exposure occurred primarily through the tective clothing began an extensive cleanup of Ihe building. The cleanup I environment. A different composition of the PCBs may be found in serum or operations lasted four years, and the cost has been enormous (12). Since then I adipose tissue samples from occupationally exposed workers. For instance, if several other transformer fires have occurred. These fires have not resulted in the workers arc primarily exposed to Aroclor 1016 or Aroclor 1242, which as much contamination as Ihe one in Binghamton, partly because the contain much less of the more highly chlorinated homologs, then their transformers in Ihe other fires were usually located in a separate.vault (13). gas-chromatographic patterns resemble a combination of Aroclor 1016 or o The problems with polybrominated biphenyls (PBBs). which arc also a o mixture of chemicals, have been quite different. In 1970 a chemical company o in Michigan manufactured polybrominated biphenyls as flame retardants. 'Ihe w same company also produced magnesium oxide, a chemical commonly mixed j ' 1242 and Aroclor 1254. For this reason Smith et al (17), in evaluating occupational exposure, divided PCBs into high and low chlorinated biphenyls. The gas-chromatographic pattern of the PCB mixture present in humans can be used to determine whether the exposure occurred primarily o HARTOLDMONOQ23643 90 KIMBROUGH through occupation or through the environment, or, in the case of occupation al situations, whether most of the exposure was recent or occurred many years ago. Although 93% of the US population eats fish, the average annual per capita consumption is small: 13 lbs. per year (6. 7). If PCBs are to be quantitated in fish, the edible pnnion rather than the whole Osh must be examined. Because of their lipophilicity, PCBs are preferentially stored in the hepatopancreas of the fish, giving erroneously high levels if the whole fish is analyzed. Sim ilarly, levels in cooked fish are lower (6). Generally, PBBs are not found in the environment because they have had less commercial use than PCBs. PBB contamination is essentially restricted to Michigan's lower peninsula. Most persons who lived in Michigan during the 1973-1974 period have low-level PBB body burdens (18). The greatest degree of contamination occurred mainly in areas with contaminated farms; this segment of the population still has appreciable body burdens (19). Since PBBs and PCBs are lipophilic, they are preferentially stored in adipose tissue. They are also present, to a smaller extent, in serum and other organs and in human milk. The concentration of these materials in different organs depends upon the lipid content of such organs, with the exception of the brain where the concentration is lower than the lipid content would indicate. PCBs and PBBs pass the placenta and ate primarily excreted through bile and milk. In addition to lipid content, the ratios between adipose tissue, blood, and vital organs are influenced by exposure level, sex, age, length of exposure, and also by whether exposure is current. At very low con centrations an analytical imprecision influences the ratios much more than at higher concentrations (19). Since human milk is relatively easy to obtain, it has been used to monitor human exposure. Jensen (20) recently summarized results of such monitoring studies. Average levels of PCBs below 2 ppm (mg/kg) in milk fat have normally been found, although women living in heavily industrialized urban areas may have higher levels. The fat concentra tion in human milk averages 2.6-4.3% (21). At 2% fat, I liter (I) of milk would contain 0.04 mg or 40 fig if the PCBs were present in milk fat at a concentration of I ppm. If an infant weighed 3 kg and imbibed 730 ml of milk per day, it would take in about 6 fig/kg, a dose that exceeds the 1.3 fig/kg dose calculated as acceptable by Cordle et al (6). At 1% milk fat this dose would be reduced to 3 fig/kg. As the infant gains weight, the dose on a kilogram body weight basis will be reduced to some extent, however; milk is the sole food source for only about six months. After the first week, the daily milk intake is estimated to be 150 ml/kg body weight per day. This consump tion gradually falls after two months and declines to 120 ml/kg body weight at four-to-six months. Finally, the amount of PCBs and other halogenated organic chemicals declines with lime. However, al low concentrations this HUMAN HEALTH EFFECTS OF PCBs & PBBs 91 may not be obvious because of continued exposure of the mother and the variability of the analytical results. In addition to PCBs, human milk contains trace amounts of many other persistent chemicals. Whether the infant's consumption of such chemicals has any adverse health effects is not known. Most persons, particularly in industrialized countries, have had some expo sure to polychlorinated biphenyls even if they do not eat fish. The con centrations at which such exposure presents a risk are not clear. Recently, Cordle et al (6, 7) calculated the dose of PCBs to people consuming fish from Lake Michigan. They concluded that persons eating Lake Michigan fish ingested an average of 46.3 mg of PCBs per year; this amount ranged from 14.17 to 114.31 mg/year/person. The calculated mean daily dose received by the exposed group was 1.7 fig/kg/day and ranged from 0.09 to 3.94 fig/kg/ day. Thus, the average spons fisherman consuming contaminated fish would receive a total PCB dose equal to 200 mg in about 4.3 years. No adverse health effects or groups of symptoms clearly related to PCB exposure could be identified in this exposed group. The presence of PCBs in the exposed persons has not caused any observable adverse health effects similar to those observed in the Yusho population (see below). However, this finding docs not exclude the possibility that the effects are loo subtle for detection or that they require long-term observation. Similarly, in Michigan an analysis of 1,075 human milk samples showed that all contained PCB residues and that the residues ranged from trace amounts to 5 ppm (mg/kg) based on fat level. The public health significance of PCB residues in human breast milk and their effects on breast-fed infants are unclear. Since there are no human data on which to base public health policy, risk predictions for PCBs have been based on results from animal studies, particularly the positive bioassay studies. Reviewing these data, Cordle et al (6, 7) concluded that a 2-ppm (mg/kg) tolerance for PCB in fish be established, since a I-ppm (mg/kg) tolerance does not greatly reduce the estimated risk. As previously mentioned, some of the isomers of the PCBs and PBBs are much more easily degraded or metabolized. Because they can be metabolized, they are more easily excreted. Others may be retained in the body for long periods; in general, the PBBs appear to be more persistent in human tissues than the PCBs (19. 22). POLYCHLORINATED BIPHENYLS Background When PCBs were first used industrially some workers developed chloracne. Results of early animal studies seemed to suggest that PCBs might have some toxic effects on the liver. Beyond that observation no information was avail- HARTOLDMON0023644 O o o CJ fj 92 KIMBROUGH able. Because PCBs were so inert chemically, they were not considered to cause a great deal of toxicity. In 1968 a poisoning outbreak occurred in Japan (23) that affected over 1,000 persons. These individuals had purchased rice oil. in large drums, from a single source and had used this rice oil for cooking. Chloracne was one of the leading signs in those who became ill. It was soon discovered that PCBs had been used as a heat-exchange fluid in the factory where the rice oil originated. PCBs had leaked out of the columns in which they were contained into the rice oil when the rice oil was heated. Since the disease was caused by ingesting contaminated rice oil. it was called Yusho (rice-oil disease). When the outbreak first occurred, its association with exposure to PCBs was not dear. At that time the capabilities for measuring these types of chemicals in tissues and body fluids were limited, particularly in Japan. Therefore, early in the investigation total chlorine, rather than PCBs. was measured. Retrospectively determining the precise dose these patients received is difficult. Whether the consumed oil was uniformly con taminated is also not clear. However, a relationship between the amount of rice oil ingested and some symptoms could be established (24). Because of this poisoning outbreak and other environmental problems, animal studies were started in Japan, in the United Slates, and in other countries to elucidate the toxic effects of PCBs. These data are summarized in several detailed reviews (16. 2S. 26). Animal Studies This article addresses primarily the human health effects of PCBs and PDBs. Therefore we highlight only recent results from animal studies that might give a better understanding of implemented public health policies and potential human health effects. One of the difficulties in using animal data to predict human health effects for PCBs and related compounds is that animal species vary greatly in their responses. Further, many of the animal studies use relatively high doses. Therefore, determining how such animal studies relate to the human situation is difficult. Some animal species, such aa the subhu man primates, the guinea pig, and the mink, are much more sensitive to the toxic effects of PCBs than the rat or the mouse; also the types of toxic effects and morphological changes in the organs of different species vary. Most animal studies conducted during the 1970s used mixtures of PCBs. In general. PCBs were found to affect reproduction and the immune response, and to cause liver tumors in rodents (16). When different mixtures of PCBs were studied, however, the results were inconsistent. For instance, the mix ture Arocior 1234 affects reproduction in rats at much lower doses than does Arocior 1260 (27). More recently some of the isomers of the PCB mixture were found to be much more toxic than others (28-32). The more toxic isomers constitute only HUMAN HEALTH EFFECTS OF PCBs Sc. PBBs 93 a very small portion of the mixture, particularly those with less chlorine by weight, such as Arocior 1242 or Arocior 1016. Recently, Schaeffer ct al (33) found that a German PCB mixture--Clophcn A-30, with an average composi tion of 1% monochlorobiphenyl, 20.7% dichlorobiphcnyl, 37.4% Irichlorobiphenyl, 17.3% letrachlorobiphenyl, 1.8% pcntachlorobiphenyl, 1.0% hexachlorobiphcnyl, 0.6% heptachlorpbiphenyl, and 0.1% octachlorobiphenyl--produced a 3% incidence of hepatocellular carcinoma, whereas Clophen A-60 produced a 61% incidence of hepatocellular carcinoma in Wistar rats. The incidence of the disease in the controls was 2%. The Clophen A-60 had an average composition of 0.2% monochlorobiphenyl. 1.1% dichlorobiphenyl, 2.2% trichlorobiphenyl. 3.1% letrachlorobiphenyl. 19.8% pcntachlorobiphenyl, 43.2% hexachlorobiphcnyl, 23.3% heptachlorobiphenyl, 4.7% octachlorobiphcnyl, and 0.3% nonachlorobiphenyl. Sim ilarly. Norback & Weltman (34) and Kimbrough et al (33) were able to produce hepatocellular carcinomas in rats with Arocior 1260, the more highly chlorinated Monsanto product. When Arocior 1234 was fed to rats, fewer liver tumors developed in exposed rats (36); however, the incidence of gastric intestinal metaplasia and adenocarcinoma of the stomach increased (37, 38). Whether PCB fractions without hexachlorobiphenyls, heptachlorobiphenyls, and octachlorobiphcnyls produce hepatocellular carcinomas in rodents should be explored. Particularly in the United States, mixtures such as Arocior 1242, 1234, and 1016 were used more than Arocior 1260. Because of these differences in potency, the PCBs in heavily contaminated areas of our environment should be characterized according to their isomeric composition. For instance, whether the PCBs in Lake Michigan arc of the same composition as those found in New Bedford Harbor, Massachusetts, is not clear. Aside from tumor formation, PCBs cause a variety of other biological effects, such as the induction of enzymes (39)j In some species they may cause atrophy of the thymus, inlrahepatic bile duct hyperplasia, hyperplasia of the epithelial lining of the urinary bladder, atrophy of the sebaceous glands, and hyperkeratosis of the ducts (40). Some isomen are feloloxic. and some produce metaplasia of the sebaceous glands, nailbeds, ameloblasts, thymus corpuscles, and gastric mucosa (28, 41). Subhuman primates, mink, and guinea pigs are particularly sensitive to the toxic effects of PCBs; other species, such as the rat, the mouse, and the dog, can tolerate much higher doses. From empirical observations, humans also appear to be less sensitive . to the toxic effects of PCBs. The ability to store these chemicals In adipose t tissue may be protective. Generally, the subhuman primates and mink have ' less adipose tissue than humans. Animals with greater ability to store vitamin A on a quantitative basis, such as the hamster and the rat, are somewhat less susceptible to the toxic effects of these types of compounds (42). The HARTOLDMON0023645 o C3 O LD 94 KIMBROUGH mechanism by which these types of chemicals affect hepatic retinoids is not clear. Apparently, the duration of the reduction of hepatic retinoids docs not correlate with (he induced aryl hydrocarbon hydroxylase (Allll) activity (43). Body Burdens Many investigators have reported PCBs in human (issues (44, 43). In the United States, according to data from the Centers for Disease Control (CDC), mean PCB serum levels ate about 5-7 ng/ml (pbb), although some patients may have higher serum levels without any documented unusual exposure. These data were also summarized by Kteiss (46). Levels in adipose tissue and in human milk fat are 100-200 times as high, since PCBs are highly lipid soluble (20). Mes et al (47) reported that PCB levels in adipose tissue of accident victims ranged from 0.9-9.4 mg/kg. ' Sahl et al (48) surveyed PCB blood levels in 738 prc-employed and 1,058 currently employed workers of a utility company. The median blood level before employment was 4 mg/I and the range, 1-37 mg/I. These levels were quite similar to (hose in (he currently employed group. Patients who died of cancer in Denmark had somewhat higher levels of PCBs in their adipose tissue (49). Since terminal cancer patients have usually lost a great deal of weight, bioconcenlration may have occurred. Similarly, in patients with highly impaired liver function, tissue concentrations of xenobi- olics may be slightly higher than those in healthy persons. However, levels remained higher if parameters such as weight, height, occupation, and resi dence were considered (50), whereas levels of PCBs in breast fat tissue from patients with breast cancer were similar to those of controls (51). Furthermore, Lawton et al (52) demonstrated that random errors and Interlaboratory variations in procedure and methods of data reporting can influence serum and adipose PCB levels. Unless an interlaboratory quality- control system is set up, measured levels between laboratories are not neces sarily comparable. For instance, Lawton et al (52) found that the results of repealed analyses on scrum samples of known composition showed the 95% prediction interval for an individual measurement to be about 42%. This interval depends on the method of extraction, the procedure used, and the means of quantitation. Summary of Human Epidemiology Studies Recently, investigators studied the predominantly black population of Triana. a small rural town in the southern United States (53). This population was excessively exposed to DDT residues by consuming contaminated fish. The residents also had PCB body burdens. Fish consumption correlated positively with PCB blood levels; no other source of PCB exposure could be established. These researchers noted that PCB serum levels increased with age and that > , j ! ` HUMAN HEALTH EFFECTS OF PCBs & PBBs 95 levels were lower in females of each age group. Similar findings were made for DDT residues. The serum cholesterol level was positively associated with the log PCB level, independent of age, sex, fish consumption, body-mass index, and alcohol consumption. Rales of borderline and definite hyperten sion for study participants were 30% higher than those expected on (he basis of national rates (54). Log PCB serum values contributed significantly to explaining (he variability of log systolic and diastolic blood pressure in multiple regression analysis (55). Median total cholesterol levels of in dividuals in the United States increase with age from about ISO to 160 mg/di at age 20 to over 200 mg/dl at age 50. PCBs in blood are influenced by serum lipid content, and populations with inherently lower total serum cholesterol levels appear to have a different PCB scrum to adipose tissue ratio. The age-associated increase in blood PCB levels could be related to (he long half-life of some PCB isomers that are preferentially retained in mammals (29); as long as exposure continues, a true steady slate between intake and excretion is never reached. Other variables affecting body burdens may be differences in metabolism with age. In the Triana studies, (he blood levels of total DDT residues also increased with age, and others have made similar observations (56. 57). Lawton et al (58) studied workers who had been exposed lo electrical-grade Arodor 1016, 1242, and/or 1254; the study covered (he period from before the workers were exposed lo (wo years after PCB exposure ceased. Serum levels for the lower chlorinated PCBs in 1977 ranged from 57-2270 ppb and in 1979, from 12-392 ppb; for (he higher chlorinated PCBs serum levels ranged from 6-142 ppb in 1977 and from 4-108 ppb in 1979. These findings again illustrate the preferential excretion of lower chlorinated PCB. Lawton et al (58) also found (hat cholesterol levels correlated with log serum PCBs. Similar associations with log scrum PCBs were found for log gamma glutamyl transpeplidase (GGTP) and, in some cases, for log alanine aminotransferase. When the PCB concentrations were expressed as levels in serum lipids, all the associations between serum lipids or enzymes and log serum PCBs disappeared except for those between log GGTP and log PCBs. Similarly, Chase et al (59) found no significant correlation between either serum triglycerides or aminotransferases and the PCB levels in adipose tissue. How age and length of exposure affect these parameters is not adequately explained in the article. Finally, Akagi & Okumura (60) were not able lo confirm a positive association between PCB blood levels and elevated blood pressure in Yusho patients. Thus, as Brown (61) has suggested, the positive association between PCB serum levels and elevated triglycerides and scrum cholesterol can be explained by the increased solubility of PCB in scrum with higher lipid content. In several cross-sectional studies of exposed workers, only minor abnormalities not necessarily related lo PCB exposure have been detected HARTOLDMON0023646 cG ^O O O 96 KIMBROUGH (62-66). In cross-sectional studies, however, the ability to evaluate chronic health effects is limited. In several studies, a positive association between results of one liver function test--the test for y-glutamyltranspeptidase--and PCB blood levels has been found. Kimbrough (67) has summarized earlier studies on the health effects of PCBs observed in workers. In 1930 and 1940. chloracne, a disfiguring skin disease, was reported among workers exposed to PCBs. One of the clinical features of chloracne is the chloracne cyst, which is skin colored and measures from I --10 mm in diameter, wilh a central opening. The other dominant lesion is the comedo. The skin lesions may only involve the face, but many also extend to other parts of the body. Microscopic examination of human skin biopsies from chloracne cases shows markedly dilated hair follicles filled with keratin. The sebaceous glands involute partially or completely. The epithelial cells lining the hair follicles and the adjacent surface epithelium proliferate, and acantho sis is present. In old lesions, the epithelial lining of the greatly dilated hair follicles becomes atrophic. Jones Sc Alden (68) examined 17 of 23 workers engaged in the production of PCBs. The workers had chloracne involving the face, genitalia, trunk, and extremities. Before the outbreak of chloracne in the plant, the electrical properly of the PCBs had fallen below specifications, and the color had deepened. In the report, symptoms of illness were extensively described for the first worker who was diagnosed as having chloracne. This worker com plained of lassitude, loss of appetite, and loss of libido. Over the years, other cases of chloracne following exposure to PCBs have been reported. Most of these involved exposure to vapors that developed when PCBs were healed (69). At times, the skin rashes that developed in workers were accompanied by pruritus. Some workers also complained of burning of the eyes, nose, and throat; dry throat; nausea; and dizziness. Meigs et al (70) reported chloracne in workers who had been exposed to PCB vapors for 5--14 months. The concentration of PCBs in the workers' breathing zone was 0.1 mg/m1. Evidence of slight liver injury was also present. Ouw el al (71) found air levels in a capacitor plant that ranged from 0.32-1.44 mg/m1 Aroclor 1242 (PCB). Here workers complained of burning eyes, face, and skin in general, and persistent body odor. One worker suffered from chloracne, five com plained of eczematous rashes, and a few had abnormal liver function tests. These workers had a mean PCB blood level of about 400 ppb (/xg/kg). In most studies of workers with chloracne, evidence of liver injury was also found; in one study, workers who did not have chloracne were found to have abnormal liver function (69). PCBs also affect the liver by inducing mixed-function oxidases. Alvares et HUMAN HEALTH EFFECTS OF PCBs A PBBs 97 al (72) determined that in five workers occupationally exposed to Aroclor 1016--a PCB mixture primarily composed of dichlnrobiphenyls, trichlorobiphenyls. tetrachlorobiphenyls, and pentachlorobiphcnyls--plasma antipyrine half-life was significantly lower than that in matched controls, suggest ing (he induction of mixed-function oxidases in the liver. These workers had been exposed to Aroclor 1016 for at least two years and had no obvious symptoms of PCB poisoning. Other health effects are eye and upper respiratory irritation. Warshaw et al (63) studied a group of 326 workers in a capacitor plant with a mean employment of more than 15 years and mean employee ages of 41.1 years for males and 47.3 years for females. Work-related eye or upper respiratory irritation was reported by 48% of the workers, and 10% had experienced tightness in the chest. Spiromelric studies were conducted on 309 workers; 66 of them were dropped from the study because (hey had been exposed to talc, textile dust, or asbestos. Thus, 243 men were available for analysis. In males, there were about twice as many smokers and exsmokers as nonsmokers. In females, the proportion of nonsmokers was higher. Thirty-four of the workers (14%) had a reduced vital capacity, and 27 of these demonstrated a restrictive pattern of impairment. Because of additional variables such as smoking and asbestos exposure, these findings arc difficult to interpret. Taylor el al (73), in an attempt to determine whether the fetus would be affected in capacitor workers, examined pregnancy outcome and birth weight, and found (hat the gestation period was reduced by one week. The infants weighed slightly less than the controls; this finding could be explained by the reduced gestation period. Smoking and alcohol consumption were not con trolled, however; furthermore, whether the socioeconomic status of this group of women was similar to that of the control group is not clear. Thus, until other studies confirm these findings, they should be viewed with caution. In several papers Jacobson and his associates reported behavioral changes and a reduced gestation period in associatinn with higher fish intake or higher intake of PCBs (9, 74-76). Furthermore, Jacobson et al (74) reported that intrauterine PCB exposure may have a delayed effect on central nervous system functioning. Since genetic makeup, the mother's lifestyle, and acute illness also affect these parameters, these findings are difficult to interpret. Furthermore, many other chemicals are also excreted in human milk (20). Kogan Sc Gladen (77). for instance, found that mothers with high levels of DDE 11,1'-(2,2-dichloroethenylidene)-bis-4-chlorobenzene| in their milk tended to wean (heir infants earlier, as they did not thrive. Apparently, PCB levels in milk were higher in older women, women who drank alcohol regularly, and primiparas (78). Whether high levels of DDE affect lactation is not dear. In animals. DDT tfl: HARTOLDMON0023647 GuO 98 KIMBROUGH homologs, but not specifically DDE, have been shown to have estrogenic effects (79). Before these findings can be clarified, additional studies must be done. No conclusive evidence thus far reported shows that occupational exposure to PCBs causes an increased incidence of cancer. Bahn el al (80) reponed results of a preliminary study of a group of SI research and development employees and 41 refinery plant employees at a New Jersey petrochemical facility. Between 1949 and 1957 these workers had been exposed to Aroclor 1254. Three melanomas and two carcinomas of the pancreas were found. This incidence was significantly higher than expected. Exposure to other chemicals also occurred, however, and the cohort was small. Brown & Jones (81) conducted a retrospective mortality study of 2.567 workers in two capacitor plants. The relatively few deaths (163) severely limited the statistical power of the study, and the average follow-up was only 15 years, whereas latency periods of 20-30 years are not uncommon for cancer. Over 50% of the sample had exposure to PCBs for two years or less. Deaths from liver cancer, cirrhosis of the liver, and rectal cancer were slightly higher than expected, but not significantly for both sites comhined. The observed increase for cancer of the rectum was statistically significant among females at one of the plants. In a follow up study (82) no additional cancers of the rectum were noted, and the standardized mortality ratio (SMR) dropped from 336 to 211. However, two additional cancers of the liver and biliary tract were observed, bringing the total of these tumors to five as reponed on the death certificates. However, a review of the medical records raises questions about at least one of these tumors. Bertazzi el al (83) reviewed the mortality of 290 males and 1.020 females who had worked for six months or more in capacitor production. Males had a statistically significant increased number of deaths from all neoplasms. When deaths were analyzed by organ system, deaths from neoplasms of the di gestive system, the peritoneum, and the lymphatic and hematopoietic tissues were higher. Among females, all causes of deaths were significantly elevated. The actual numbers in this study, however, were small. Yusho and Yucheng Two outbreaks of poisoning have been reported that followed the ingestion of rice oil contaminated with polychlorinated dibenzofurans, biphenyls, and quaterphenyls (PCQs). The first outbreak occurred in Japan in the summer of 1968 and the second outbreak, in Taiwan in 1979. Ironically, the outbreak in Taiwan repealed what had occurred 10 years earlier in Japan. Many studies of these two outbreaks have been published in Japanese or Chinese. In 1984 some of the information in these reports was published in English in the ':o o HUMAN HEALTH EFFECTS OF PCBs & PBBs 99 American Journal of Industrial Medicine 5:1-153; the information is also summarized in volumes 59 and 60 of Environmental Health Perspectives. In Japan and Taiwan the disease was first recognized because chloracne developed in the affected patients (84). In Japan, members of all of the affected households had purchased rice oil from a specific company, and the toxic rice oil produced or shipped on February 5 and 6 of 1968 contained large amounts of Kancchlor 400, a brand of PCB with a chlorine content of 48%. At the time of the outbreak, no analytical methods specific for PCBs were available in Japan; the concentration of Kancchlor 400 in the oil was therefore estimated from the organic chlorine content to be 2,000-3,000 ppm. Kanechlor 400 had been used for heating the rice oil in a metal container at over 200C. at a reduced pressure of 3-44 mm llg, to remove odorous material from it. Kancchlor (K) must have leaked from the heating pipe into the processed oil, but the actual mechanism of the contamination has apparently not been determined. The rcanalysis of the K-rice oil. once the methods were developed, showed that some of the oil samples contained 1.000 ppm (mg/kg) PCB. This concentration was much lower than had originally been estimated. Therefore, other chlorine-containing compounds were assumed to be in the oil. and additional samples were analyzed. The oil was found to contain an average of 5-ppm polychlorinated dibenzofurans (85). According to Buser ct al (86). the Yusho oil contained more than 40 polychlorinated dibenzofuran isomers, including the highly toxic 2,3,7,8-tetrachlorodibenzofuran (TCDF) and 2.3,4.7.8-pentachlorodibenzofuran (PCDF). In addition, the oil con tained PCQs at a concentration of 866 ppm (87, 88). According to estimates made by Kuratsune (89), the total amount of PCB, PCDF. and PCQs consumed by the patients was. on the average. 633 mg of PCB. 3.4 mg of PCDF. and 596 mg of PCQ. This calculates to roughly 157 /ig/kg body weight/d. PCB. 0.9 Mg^g body weight/d. PCDF, and 148 #ig/kg body weight/d. PCQ. At this dose the length of the latent period between exposure and onset of clinical illness was roughly 71 days, with a range from 20 to 190 days. Some of the oil the patients consumed may have contained higher or lower levels because in such situations contamination is usually not uniform. Furthermore, the patients consumed different amounts of con taminated rice oil. The severity of symptoms was positively associated with the amount of contaminated rice oil consumed (24). Early in the outbreak the patients had chloracne. dark-brown pigmentation of the nails, itching, pigmentation of the skin, swelling of the limbs, pig mented mucous membranes, eye discharge, hyperemic conjunclivae, jaun dice. swelling of the upper eyelids, a feeling of weakness, numbness of the limbs, and fever. Over 1.000 people were affected. Thirty-six babies showed fetal PCB syndrome, which consists primarily of a dark-brown pigmentation HARTOLDMON0023648 100 KIMBROUGH of (he skin (Cola babies). The cutaneous pigmentation was caused by an increase in melanin pigment in the epidermis (90). The mucous membranes were also pigmented. In all cases, the pigmentation disappeared by the lime the babies were between two and five months old. In affected infants, the face was edematous, and spotty calcifications were noticed in the parietal and occipital areas of the skull. In a few of the infants, the teeth had erupted at birth. Subsequently, the adult patients with clinical disease complained of having to expectorate a great deal and. on auscultation, wheezing was noted; however, on examination, there was no evidence of bronchial asthma or pulmonary emphysema. In many of these patients, the respiratory symptoms have pcrsisled. and the patients have chronically infecied airways. In the early 1970s. some changes were noled in the patients' serum immunoglobulin levels, bul the levels relumed lo normal. Over lime Ihe severity and the extern of ihe skin lesions improved considerably in the exposed population. Fifteen yean after the accidenl, only a very few patients had exlensive chloracnc (91). About five yean after the oulbreak of Yusho, tissue and body fluids of Yusho patients were analyzed for various congenen of PCB and PCDF. At this lime. Ihe PCB levels in adipose lissue were 1.9 i 1.4 ppm (mg/kg). In Ihe liver Ihey were 0.08 0 06 ppm and in blood, 6.7 5.3 ppb (/ig/kg); thus, they were not very different from levels in the general population in Japan. On the other hand, the isomeric distribution for the PCBs in the Yusho patients varied from that in the control population in the same area (92). About 40 PCDF congeners were identified in Ihe rice oil that the Yusho patients ingested- Only some PCDF congeners were retained in the body for a long lime; they included 2,3,6,8-TCDF, 2.3,7,8-TCDF, 1,2.4,7,8-PCDF, 2.3.4,7,8-PCDF, and 1,2,3,4,7.8-hexachlorinated dibenzofurans. Since these congeners do not have free adjacent carbon atoms, they are not as easily metabolized and excreted. More of the 2,3,4.7.8-PCDF than Ihe other iso mers was retained in Ihe patients' tissues. In the five palients studied, the concentration of this isomer ranged from 6.9 ppb (/ig/kg) in a specimen obtained in 1969 lo 0.1 ppb (/ig/kg) in a specimen collecled in 1977. Measurable concentrations of TCDFs were only detected in Ihe earlier years. Although not the most toxic isomer. Ihe 2.3.4,7,8-PCDF caused mixedfunction oxidase induction at a dose of I /*g/kg in rats, and atrophy of the Ihymus, suggesting toxicily at a very low dosage level. Thus, Ihe clinical o manifestations observed in these patienls were primarily caused hy the PCDFs. specifically by the more toxic isomers. In the Yucheng episode, il was never determined with certainty how the o rice oil was contaminated (93). In 1979 a school for blind persons informed a local health bureau in Taichung Counly lhal a strange disease characterized by an acnelike skin eruption had been occurring frequently among students and 4 X? HUMAN HEALTH EFFECTS OF PCBs & PBBs 101 staff since the end of March. At the same lime. 85 of 150 workers in a nearby plastic shoe factory had Ihe same symptoms. Later that year, this outbreak was also reported to a local health bureau. Victims in both outbreaks had consumed the same brand of cooking rice oil. which had been manufactured by the same company and which had been purchased in the same store. For this reason the rice oil was the prime suspect in ihe oulbreak. In additional reports of outbreaks in other companies and in the general population, all victims had consumed the same type of C-rice oil. Finally, because Ihe disease resembled the Yusho disease in Japan, samples of C-rice oil and patients' blood were analyzed in Japan and were found lo contain either a Kanechlor-400 or a Kanechlor-500 mixture at concentrations as high as 65 and 108 ppm (mg/kg), respectively. Over 2,000 patients were finally identified as having been poisoned by contaminated rice oil. Oil samples collecled from other outbreaks contained PCBs at concentrations of 31-300 ppm (mg/kg). Retrospective studies determined lhal ihe period of PCB inlake ranged from 3 lo 9 months. The average total intake for each person varied from 0.77 to 1.8 mg of PCB. Within the first year of the outbreak, the blood levels of PCB in 13 patients ranged from 3 ppb lo 1,156 ppb. Most of the patienls had blood levels between 11 and 150 ppb (/ig/kg). The symptoms observed in these patients were quite similar to those already described for the patients in Ihe 1968 Yusho oulbreak in Japan. The rice oil was not only contaminated with PCBs but also with PCDFs and polychlorinated quaierphenyls. It contained the same major components of PCDFs observed in the rice oil in Japan--namely. 2,3.4.6,7-PCDF and 2.3.4.7.8- PCDF. Relatively high concentrations of 2.3.4.5.3',4'-hexachlorohiphenyl were found in Ihe blood and adipose tissue of the Yucheng palients. This particular PCB isomer is biologically quite active, and the concentration of 2,3,4,3'4l-pentachlorobiphenyl was also elevated in these patienls. Furthermore, as in the Yusho palients. Ihe concentration of 2.3.7.8TCDF was comparatively low (92). Chen e( al (94) analyzed additional samples of ihe oil, blood, and adipose lissue of the Yucheng palients. These investigators identified several TCDF and PCDF isomers. Apparently. 2.3.7.8- TCDF was only a minor component in the oil; the major component was 2.3.4,8-TCDF. One of Ihe major furans in Ihe toxic oil was 2.3.4,7.8PCDF. Ihe concentrations of the PCDFs in different oil samples ranged from 0.21 10 1.68 ppm. Polychlorinated quaierphenyls were present in concentrations ranging from 25 to S3 ppm (mg/kg). Overall, the concentrations of PCDFs and PCQs were lower in these oil samples than in the oil samples that had caused Ihe Yusho oulbreak. Whether these oil samples were representative is not really known. The 3.4,3'4'-telrachlorobiphenyl was also identified in the 011 lhal caused Yucheng disease in Taiwan. This isomer is considered to be HARTOLDMON0023649 O oo ID Tb 102 KIMBROUGH he most loxic PCB isomer present in commercial PCB preparations (95); it was present at a concentration of about 1%. Most other commercial PCB preparations, such as the Aroclors. in the United States have not been shown to contain this particular isomer. In addition to the epidemiological studies, some disease-specific in vestigations were also conducted. The blood pressure of the Yucheng and Yusho patients was not affected (60). Although some of the patients in the Yusho cohort have died of cancer (90), the number has been small; because the latency period may be long, the population should be followed for a longer period to determine whether Ihe cancer incidence will increase. Although PCBs and related compounds are known to affect reproduction in animals, and although they affected some fetuses and neonates in the Yusho and Yucheng episodes, the information on reproduction and fetal toxicity in general is very limited. In one such study, Hara (96) examined women working in a capacitor plant who also nursed their infants and who themselves had mild chloracne and erythema of the skin. The human milk of some of these women contained, on a whole milk basis. PCB levels that ranged from below 50 ppb (/xg'kg) to about 400 ppb (/xg/kg). Forty children of these mothers were followed for a five-year period. Some children were found to have "decayed" nails, gingival pigmentation, mottled enamel, and denial caries. No relationships between these changes or symptoms to PCB blood levels, however, were observed. The general population in the United States and other countries also has body burdens of PCBs, PCDFs. and polychlorin ated dibenzodioxins (97, 98). However, these background concentrations-- particularly for the biologically active isomers--are far lower than they were in the Yusho and Yucheng patients, even several years after exposure. Chang el al (99) examined the delayed immune response in 30 Yucheng patients and compared their responses with those of 50 controls. The mean age of patients in both groups was about 14 years. The authors injected a solution of streptokinase and streplodomasc subcutaneously into the flexor side of the forearm. The response was read al 24 hours (hr) and again at 48 hr after injection. Eighty percent of Ihe controls had an induration of 5 mm or more in diameter 24 or 48 hr after they were injected; only 43% of the exposed group responded similarly. All of the poisoned patients had dermal lesions, and Ihe percentage of patients with a positive response decreased with increasing severity of the skin lesions (chloracne). Furthermore, the degree of the dermal lesions appeared to be associated with the whole blood PCB concentrations. Patients with minor skin lesions that were classified as grade I appeared to have a normal skin response. The same authors found that PCBs caused a decreased concentration of IgA and IgM. but not of IgG, in serum. Furthermore, the percentages of total T cells, active T cells, and T mu cells decreased, whereas Ihe percentage of B cells and T gamma cells were not HUMAN HEALTH EFFECTS OF PCBs & PBBs 103 affected (100). These two reports are the first in which the effect on the immune response was actually correlated with body burdens of PCBs and in which only severely poisoned patients showed this effect. This finding is consistent with the findings from animal studies in which relatively high doses of PCBs affected the immune response and also caused some other adverse effects. In the Japanese and Ihe Taiwanese Yusho and Yucheng poisoning out breaks, sensory neuropathy was reported in a number of patients for whom nerve conduction velocities were measured (101, 102). The blood levels of Ihe various chemicals (PCBs. PCDFs, PCQs) were negatively correlated with the lowered nerve conduction velocity, suggesting that these types of chemi cals affect nerve conduction velocity. (If is not quite clear why most in vestigators measure nerve conduction velocity to detect sensory neuropathy. Other tests that would measure the detection of vibration, touch, and tempera ture would be more useful from a clinical perspective.) Seppalainen el al (103) examined 16 men working in a cardboard plant who were exposed to fumes that resulted from the explosion of 15 capacitors containing Clophen A-30. The first PCB air concentrations, measured 5.5 hr after the explosion, were 8,000 to 16,000 /xg/m3 air. PCDFs were also formed. The soot samples contained tetrachlorodibenzofuran up to 90 /xg/g. of which 6.5 /xg/g was 2,3.7.8-tetrachlorodibenzofuran. In addition, monochloropyrenes and dichloropyrenes were found. Most of the men had a transient sensory neuropathy in their lower extremities. Chang et al (104) reported increased urinary 5-aminolevulinic acid uropor phyrin excretion in 69 Yucheng patients over that of 20 controls. No informa tion on the patients' clinical conditions or on how these findings related to degree of exposure was given. No such observations have been reported from Japan. POLYBROMINATED BIPHENYLS Since the toxicity of PBBs both in laboratory animals and livestock was recently reviewed (105), we do not review it here in detail. In laboratory animals, PBBs generally cause effects similar to those that the PCBs cause. They produce morphological changes in the liver, affect reproduction, and promote biochemical changes, such as hepatic porphyria and induction of mixed-function oxidases. Teratogenic effects have also been noted. In addi tion, atrophy of the thymus has been reported, and hepatocellular carcinomas have been produced in both rats and mice. The overall findings reported in animal studies are similar to those that have been reported for PCBs. Although the PCB contamination of the environment is a more general problem, the PBB contamination primarily affects certain areas within the state of Michigan. Most persons living within the lower peninsula of Michi- HARTOLDMON0023650 104 KIMBROUGH gan have had slight exposure, since the contamination resulted from dairy products and since normal marketing channels for these products involved the mixing of milk from many producers in relatively few processing facilities. In addition, most cull dairy cattle are used for hamburgers and processed meat products that would also receive wide distribution. Thus, the marketing system diluted the degree of exposure for the individual; however, it increased the number of those exposed. In 1978, the distribution of PBBs was com prehensively studied in a probability sample of 1.738 persons. PBB levels in scrum were determined, and 844 adipose tissue samples were also analyzed for PBBs. PBBs were detected in 97.3% of the adipose tissue samples, in 68% of the adult serum samples, and in 72.7% of the serum samples from children. The mean PBB concentration in adipose tissue was 400 pph (/ig/kg); in scrum it was 1.3 ppb (/ig/kg) for adults and 1.8 ppb (ju/gkg) for children. The highest adipose tissue concentration was 37 ppm (mg/kg) (18). In addi tional studies, when cohorts of PBB-exposcd residents of Michigan were compared with residents of the stale of Wisconsin, a higher prevalence of a variety of symptoms and complaints was noted in the Michigan residents (106). Similarly, in comparative neurobchavioral studies, the Michigan pop ulation was found to be affected more than that in Wisconsin (107). Since the findings were not correlated with body burdens of PBB in any of these studies, determining whether other factors may be responsible for these differences is difficult. In 1976, the Michigan Department of Public Health established a cohort of farmers who had been exposed to varying con centrations of PBBs in their products and their environment. A total of 3.877 persons were enrolled. They included farm residents, direct recipients of farm products, chemical workers and their families, and a few persons who had been originally studied in a smaller previous study. The serum PBB levels in this entire group ranged from no detectable levels to 1,900 ppb (^tg/l). with a mean of 21.2 ppb (jig/I) and a median of 3 ppb (jig/I). Because of the wide range of exposure and because results could be analyzed by regression analyses with exposure as a variable, a comparison group for acute health effects was not included. This cohort was found to have various symptoms and conditions; however, these symptoms did not correlate with PBB body burdens. Symptom prevalence rates were slightly higher in persons with no detectable PBBs in serum than in those with measurable quantities. In all groups, including chemical workers and quarantined farm residents, the highest prevalence rates were in persons with the lowest serum PBB levels (22). Similarly, in this study and in a previous immunologic study (108) no dose-related depression of lymphocyte function in persons exposed to PBBs could be demonstrated. All these findings suggest that there may be no causal HUMAN HEALTH EFFECTS OF PCBs & PBBs 105 relationship between the abnormal lymphocyte functions observed in some persons or the prevalence of other symptoms and exposure to PBBs. This cohort of Michigan residents is still being followed by the Michigan Depart ment of Health in collaboration with the Centers for Disease Control. Several studies of subgroups of this population pnd surveys for chronic health effects have been conducted since the cohort was First assembled (109, 19). When serum and adipose tissue concentrations were compared, a significant correla tion was found. The serum: adipose tissue concentration ratios ranged from I to 140 to I to 260 for pregnant women and male chemical workers, respec tively. Males from farms had a significantly different ratio of I to 325 to 329. Potential transplacental passage of PBBs was demonstrated, since they could also be found in the fetus and newborn. Cord blood contained one-tenth of the concentration found in the maternal serum, which indicated partial placental passage. Human milk contained PBBs at 107-119 times the quantity found in maternal serum. PBBs were also delected in bile and feces, which indicates that these materials can be transferred into the intestinal tract. All of these concentrations were measured long after the population bad first been exposed to PBBs (19). Concentrations of PBBs observed in bile and feces were about one half to seven-tenths of the serum levels and are probably about 0.5% of the adiposejissue levels. These findings indicate lhai PBBs are very slowly excreted, which is consistent with the Findings of Tuey & Matthews in rats (110). The estimated half-life for PBB is 6.5 years. More recently, two groups of Michigan residents--those with high PBB serum levels and those with PBB serum levels around I ppb--were matched for age. sex, and smoking. For both groups, various clinical laboratory rests were conducted, blood pressure was measured, and height and weight were determined. In this study. 83 participants had PBB serum levels of 50 ppb (/x.g/1) or more. In the middle group, 83 had PBB levels of 5-49 ppb and 96 had PBB levels of 0-44 ppb (^tg/l) in serum. Urinary porphyrins were also measured in all of the participants. Thus far. the Final results of this study have not been reported. For most of the parameters studied--which included serum glucose, triglycerides, high-density lipoproteins, various liver func tions, creatinine, uric acid, thyroid function, proteins, calcium and phospho rus in serum, and also measurement of various porphyrins--no differences of clinical significance were found among different groups (M. Barone, personal communication). (Note: Even though significantly more women in the high PBB group used birth-control pills than women in (he low PBB group, too few were using them to affect urine porphyrin levels.) In none of a variety of other studies conducted on (his population as well as other groups in Michigan did any Findings indicate that exposure to PBB had impaired the health of the exposed group. All of these studies have been reviewed by Fries (105). HARTOLDMON0023651 106 KIMBROUGH Allhough this population was exposed during a 9-monlh period in 1973 and 1974. whether ii will have chronic health effects is unknown. This particular cohort needs lo be followed for 30 lo 40 years before the question of chronic health effects can be intelligently addressed. Two problems with assessing chronic healih effects are that the cohort, in spite of iis size, is slid relatively small and lhat ihe amount of exposure it has received varied widely. Although some members of Ihe group exposed lo PBBs have relatively high body burdens, these burdens are still appreciably lower than those of rats in which liver cancer developed. In the study by Kimbrough el al (111), liver cancer developed in the rats lhal received a dose of 1,000 mg/kg body weight. This dose for humans would roughly translate into a dose of 70 grams per person. These amounts are much greater than the estimated mean total exposure per person. The highest exposure was about 11.7 grams, and Ihe mean was 170 mg per person. In rats given 200 mg/kg. a dose lhal for humans would be between 12 and 14 grams, only neoplastic nodules developed in their livers: there was no evidence of hepatocellular carcinomas. Of course, whether humans would be more or less susceptible lo the toxic effects of PBBs and whether their response would be similar to that of rats is not known. In conclusion, various toxic effects of PBBs and PCBs have been described in laboratory animals. In humans, acuie poisoning outbreaks have only occurred following exposure lo a combination of PCBs and PCDFs. When humans were exposed only lo PCBs or PBBs. Ihe only observed acute effects have generally been minor. So far. no significant chronic healih effects have been causally associated with exposure lo PCBs or PBBs. Use of trade names is for identification only and does not constitute endorsement by Ihe Public Healih Service or Ihe US Department of Health and Human Services. Literature Cited 1. Jensen. S. 1966 Report of a new chernicl hazard. New S* i. 32:612 2. 19X2. Polychlorinated biphenyls (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in electrical equipment. Fed, Mrgin. 47:37342-6(1 3. |982. Polychlorinated biphenyls (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in closed and controlled waste man* ufacturing processes. Fed. Regal, 47:469X0-96 4 McLeod. K. 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