Document yk5q7ex9b5Jb7mn97KOpXew3E

*t IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION 1 CECIL SCOTT, ET AL v. MONSANTO COMPANY ] 1 No. B-84-1103-CA 1 EXHIBITS TO VIDEOTAPE DEPOSITION OF DR. GEORGE LEVINSKAS May 28, 1987 1300 Post Oak Boulevard Houston, Texas Wanda G. Kuhn, Court Reporter Nell McCollum & Associates Inc. 2900 Smith, Suite 104 Houston, Texas 77006 (713) 523-3767 - NELL MC CALLUM & ASSOCIATES, INC. STLCOPCB4028791 QMtrt DMEH Toxicology Section/St. Louis rcojoivjoerrjuocATiON) ! ' SPECIAL (TYPE OF REPORT) REPORT urs-jj rH CH/D cm a! CM o a-ct-o> a_i ur a < fit/.*i Xaoaq.. uo. a. l/l cH > J43 oo U>3 uJ _J Om X < 0.0 OH 4M3 S- O ' J- t- 'O o U43 CJI x C**M a Wao: 2 ui < CM O o CM -j to s o z o z HaU.i > & cc u n-iosaiBi REPORT NO.: MSL-2002 JOB/PROJECT NO.: OATE: October 14, 1981 TITLE: TOXICITY OF AROCLOR PRODUCTS 1242 ,;. 1254 AND 12 60 TO THE LIVER OF ALBINO-RATS ' authors: George J. Levinskas, Ph.D. A3STRact: 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. SCH 058930 000007 ^ STLCOPCB4028792 DISTRIBUTION COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - TDM EH 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-10SS<C) (RCV. 1/80) GOQCG`3ch 05893 STLCOPCB4028793 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 --* '- - i- - 0000.30 SCH 0539 32 . . STLCOPCB4028794 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- - 000C30 053933 SCH STLCOPCB4028795 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). OOQ-Ol 3 SCM 05893** STLCOPCB4028796 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 0 '1QG02 SCM 058935 ^ STLCOPCB4028797 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 STLCOPCB4028798 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(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 c *6- . GOQCO-l - $CM 058937 STLCOPCB4028799 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. SCH 058938 STLCOPCB4028800 Footnotes 1. 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. 2. R.D. Kimbrough, personal communication. 3. 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. . 4. Dr. David Ford of Bath University. . 5. 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. _ .' 6. 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. 7. Charles River Breeding Laboratories, North Wilmington, Massachusetts. 8. 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- - ooor.oG SC* STLCOPCB4028801 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 9- SCH 058940 STLCOPCB4028802 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. 00QC03 SCM 0589*1 STLCOPCB4028803 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. 000003 SCM 0539^2 ^ STLCOPCB4028804 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 \ Terrains 1 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 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 11 1 100 023 11 1 533 000 000 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". 1 b Includes 1 animal without record of examination in original IBT report. Not listed as hepatoma in worksheets for ni/> 2 ^ * iaJ O Gordon and Richter report but appears and was crossed out on typed tabulation for animal marked "Extra". for other 2 animals do not appear in records of examinations for original IBT report. Ov/> O'- ' ' GCD 0C Diagnosis does not appear in records of examinations for original IBT report. Lhi Diagnoses STLCOPCB4028805 Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254: Primary Liver Lesions Among Albino Rats 3 Honths 0 1 10 100 6 Honths 0 1 10 100 12 Honths 0 1 10 100 Tenaina 1 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 26 179 131 120 034 103 563 000 000 13 1 1 14 14 4 4b 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 wDc 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 '' STLCOPCB4028806 Sacrifice Interval Diet Level (ppm) Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats 3 Honths 0 1 10 100 6 Honths 0 1 10 100 12 Honths 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 Focal necrosis - Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia Hepatoma Cholangiohepatoma 202 3 0 0 4C 0 000 0 000 2 000 0 00 1 0 000 0 000 0 112 6 1 0 ld 0 022 0 000 3 000 0 000 0 000 0 000 0 1 25 000 000 000 000 1 10 000 000 3 0 0 4 1 2 0 0 157 9 143 6 10 1 0 0 3 13 9 107 6 5 6 7 12 0 0 le ?f,8 000 A8.h *.Control group has animal dead at 19 montha and 2 marked "Extra". 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 aniauils dead at 22, 22 months. b Includes 1 marked "Extra". c Vorkaheets show listings under this diagnosis with arrow pointing to Vacuolar change column. d ,Date of death of thia 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 2 animals without record of examination for original IBT report. Diagnoses for other 5 animals do not appear in ^ Q records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter S reports. o 08 Includes one animal with both diagnoses. Hepatoma is 1 of 2 crossed out (superscript f). n' D O Includes 3 animals without record of examinationfor original IBT report. Diagnosis for other animal does not appear in t. records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter n reports. > ^' STLCOPCB4028807 V E X H I B I T # ( I STLCOPCB4028808 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 ' - ^ DEPOSITION I EXHIBIT |cevnosicfls- / . 1 5-Q1-V7 vK. * ,,..--unmur STLCOPCB4028809 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 8 0; 5) Any document stating, reflecting or referring to the current standards, procedures and policies of Monsanto's Toxicology Department; 000002 -2STLCOPCB4028810 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, 000C03 -3- STLCOPCB4028811 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. 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. -4- OOQOO* STLCOPCB4028812 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 -5- G0QC05 STLCOPCB4028813 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 6551 Michael A. Pohl -6- OOQCCG STLCOPCB4028814 a ixx STLCOPCB4028815 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 <& Primm, 4700 Texas Commerce Tower, Houston, Texas 77002 Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt & 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 000007 STLCOPCB4028816 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. Keplinqer. 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 G0QCG3 STLCOPCB4028817 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. Keplinger. 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. Keplinger, 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- ' 000003 STLCOPCB4028818 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- - oock;:o STLCOPCB4028819 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 Floor Houston, Texas 77056 (713) 623-8800 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- 000011 STLCOPCB4028820 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 jnessenger 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 & Primrn 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 -6- ococ::r STLCOPCB4028821 V E X H I B I T STLCOPCB4028822 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 f EXHIBIT I S'-3-7- 7 STLCOPCB4028823 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- 000014 ^ STLCOPCB4028824 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 anyoutside/independent testing laboratory in gathering, recording, maintaining and/or 000015 -3- STLCOPCB4028825 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; Q0Q01G -4- STLCOPCB4028826 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. 000C17 STLCOPCB4028827 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 0000:3 STLCOPCB4028828 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 -7- 000C13 STLCOPCB4028829 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 6551 Michael A. Pohl oooc;:o STLCOPCB4028830 ix m \ STLCOPCB4028831 T*l rU VDI: 650 263 071? <409)838-9941 BESSS, MUBIN 9 SUNN LAWTEBS 909 Laurel Beaumont, Texa* 77701 Aarvnbtct: Ha JBLAHXS Ha ID: 263 071? May 13, 1967 Walter Crawford WELLS, PEYTOH, BEARD, ICEBERG, HUHT & CRAVTORD 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 55 products. 1 also understand that third parties may have done some 350 study validations on IBT studies of about 50 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 350 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. 5 DEPOSITION f EXHIBIT *Le\/tu$KPrs.-y I r-0.-7.y7 uofc 000C21 STLCOPCB4028832 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-: STLCOPCB4028833 !I ! E X H I B I T STLCOPCB4028834 Monsanto r*a * * *t wjCCT , <<<% . TO : M George Rough, Jr., M. D. September 17, 1975 ) * ^3 cJlm -Pt*r~ -ms- r</s MONTHLY COMMENTS - MEDICAL DEPARTMENT CONFIDENTIAL Corporate Administrative Committee 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 D. ^ DEPOSITION | EXHIBIT S' SCM 019722 G00G33 STLCOPCB4028835 ix m STLCOPCB4028836 ^Monsanto ' r*oa ft lOCAftat Medical Department A2SA October 13*, 1971 AROCLC^ 1260 MfOCWCC TO FILE <e R. K. Kelly M. N. Johnson B. P. Wheeler I spoke with: Tel: Renate D. Kimbrough, M.D. Pathologist EPA Atlanta Tox. Branch 4770 Buford Highway Chamblee, Georgia 303&1 (404) 633-3311, ext. 5218 today regarding her observation of bladder tumors in rata fed AROCLORl 1260. The observations of Vos and Koeaan (Toxicol, and Applied Pharmacol. 17, 656-668, 1970) on polychlorinated biphenyls led them to undertake rat reproduction studies with - AROCLOR 1254 (Lot AK38) and 1260 (Lot AK3). The materials were obtained from Glasgow at FDA. Their primary findings - have been lesions in the liver, and Dr. Kimbrough plans bo present a paper on the liver lesions at the 1972 spring meeting of the Society of Toxicology. In addition, she has seen 2 lesions in the bladders of rats fed AROCLOR 1260. The first occurred in a female which died after 6 months on test. This was diagnosed as a malignant anaplastic carcinoma of the bladder. The second was in a male killed after 8 months on teat. The first diagnosis was of epithelial hyperplasia. Sections of both bladders were sent to NCI for diagnosis to Drs. Strauss (7) and Katherine Snell. The diagnosis of carcinoma in the female was confirmed. The lesion in the male was 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. iM. aiv <. Dr. Kimbrough said that they had observed porphyria in some rata with both compounds. ' She also Indicated that they were contem plating doing a 2 year feeding study with larger numbers of rata 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 Weiss burg learned abo^ut them. \.cont a.j. DEPOSITION exhibit y^V'VSK/Kc- fr s-n-s > 00tr 10002 STLCOPCB4028837 FILS 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. I 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 Drs. Kelly and Johnson for their review /bks > SCK 0183 STLCOPCB4028838 LUXX ( I B I T / STLCOPCB4028839 October 28, 1971 Dr. Moreno L. Keplinger Industrial Bio-Teat Laboratories, Inc 1810 Frontage Road Horthbrcnk, Illinois 60062 Dear Kep: I spoke with Dr. Renata Kimbrough 0f the S?A la Chaablee, Georgia regarding our proposed visit to her to discuss bladder pathology la rets fed this product. I Indicated . to her that we expected to have all of the available bladders examined by the latter part of Voveaber, 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 Don turns up which say be of interest. This was agreeable to her. She suggested that we sight bring extra slides for referral to MCI, 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 e-ppleCof dates that would be convenient for a trip to Chaablee so that I say schedule a visit there. Xindesfc personal regards. George J. Levlnskas, Ph. B Mgr., Product Svaluatlon JL/bics cc: Dp. Don Gordon Mr. V. B. Papsgeorge Mr. 1. P. Wheeler 2 DEPOSITION EXHIBIT S~-17- f~7 oOC SCM 023586 00002G STLCOPCB4028840 STLCOPCB4028841 STLCOPCB4028842 UJXX I B II T I t STLCOPCB4028843 ,m . - -=* *- /W^KN rvJ ...................... Tw* :?>*S^ST *./ .. .- :. 3b?k: 722!&*s rW' - -?" * * Tha final TVruw^'TotJ'ithorpub'Uffa.tlan way Sary" silrJ'tly ; dopendin - . in Its noonine and; ha'rt.liaatodd provided.descriptive tcraa for tboaa' .' -: leaioos not linvin<t;tl;:elittsie opimarMm deliver, cnrcincan..'However, : .. corwJdarlnrt tli'sl^iricMCo';oClt^^fl^fc^.| (kiflM-'dlscrcte nodules" ', ~'~ and "trnhcculnr (bosnpfallic) hypnrplasln1* aaTimcaBaiVous lcsfono'and' * '"' 'A ' '.-. tints lma.-sUvo of ctaolnoi^aie msponso, ,, ttcHlw'hawj you called - . . : -~ sized am tmpal3ooli.ui.ax wltii-pscuioglacdulsr fursntioo. .< 'r- - . _ . '' ' . , p. You lnrw oftha Jfrit Idvnr Worhshoit i`aa~bavin/: oo thii typo of, V- lesion on ro`*nil'^r 11-JJUi. and wish to.llonlt you for the use of your . , notorial,' o iosiona will bd'Jdcntirlcii'viUi.j(n7 "epoeiflc coapouad, V-? " ' ` > . c now or la Uw future vjn factj'-'l^iwloctod tho'illdos blindly to'represent' fv ' .tho apoctrusof lesions and could not ldaatify than with an acmrt cyolf, ' '= c.- 5^. C:sr-fl: m inviting aU/altod nuabcr.;pf. guusta,' nod"would bo.pleased to ; 11 hove you attend,'"if you like A. It la twine bold lA iho.Ehcmtoa Motor '' ' ') Hotel in. silver Spring,' frew .H:30'n'in,';' Doccslwr. 11- "`13:00 mn .- ;-- '' ' ','.i.; Deccntor 13th. I cnnnoi'puy.ajty of yuur oxpauoai,''since you will not ", _ -' ,'V-bO a official .1 7. ' . _ ' ;. s Cnll.aieII* you want`:to di.-.cusa anythin^'-further. '.-I^sa eorxy for ,&<:; '"" UmT tlno on thin Aroclor study,'iiuit iqr.solodulu nut oblleatloM bom' .' bocooaa quite ovorvtKiiiatne.' Ao soon oa ttw cpcelolliitains UV oonplatrd, *f-.' I should ho'able'.to write tin pathal^y doccrlpticTC: and lainilations'. .1 t--- = :'' ."*" &:>' r-?: " .r . sra-- r '-v.. . _ STLCOPCB4028844 ' - SMS'..- "';; *; v , V *-*- = *.-. `m ;-.'Cb:t f",r''i-i 7 *- . ' `.Ur rinary bl_o_n_--dIeirt:.'. .ic__<1$> K\ 11__ ' ' I /l?/*:t`s/.fjf'~/ 'z'2'Asf/-l -:--7 /!a 1 r^' J- '. ` ` .J>..*';.. . ' ' STLCOPCB4028845 z rx m \ \ B I T # / / STLCOPCB4028846 F.R,Johnnnsen - A2SC December 6, 1974 AROCLOR 1254 n |*74 E. P. Wheeler - A2SA W. B. Papageorge - E2CK G. Roush A2SA In a recent lphone conversation between Dr. R. Kimbrough (CDO) and G. J. Lcvinskas, Dr. Kimbrough expressed the belief that NCI had recently contracted out a carcinogenicity study involving AROCLOR 1254. Consequently, I made a call to Dr. Sidney Siegel at NCI. Dr. Siegel indicated that he had no knowledge of an outside contract with AROCLOR 1254 and further stated he doubted there was one. He did tell me that AROCLOR 1254 has been under test at NCI in their bicassay operation program since September 1972. He reported that the experimental part of the study had "been completed as_.of October, 1974. His expectations are that the necessary histopatholoy-~wil2""be completed within the next 6 months, at which time a meeting could be set up for discussion- oSrtTlB test results. ----------------------- .- A quick check of the April 3, 1974 list of chemicals being tested . under the NCI carcinogenesis program did not reveal that AROCLOR 1234 was on test. However, the RCI 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. / c^(m) ' ' / Frederick R. Johannsen ;V C/6a J<tX+ ^ *> C*--1 ^ Jfju. rfcl/oVS y ^ 4 ooocgo STLCOPCB4028847 u ixx i B I T STLCOPCB4028848 \ OIPARTMSNT Of HIALTH. IDUCATtON.ANO WtLfARI PUBLIC HtALTM SSJtVICK - esw-rin poa wham cowtsol ATLANTA, atOBOIA NUl December 6, 1574 i i!:? Dr. Georg* Lenasksa Honsanto Industrial Cfcsal cals Cospasy 800 I. Lindbergh Boula-rard St. Louis Missouri 63164 Dear Georg*: I sm anclosing a brlaf outline of the experiment Aroclor 12-72-4 . for jour information. A copy of lob Squires latter is also anclosed.' Sinesrely yours, . Enclosure lenata D. Kiafcrough, M.D. Toxicology Branch c. --y SCM 2ZSlZ * --DEPOSITION 1 I f EXHIBIT 1 |6V/juS>C7K> - )l I i ^*^7- y7 gjt: | STLCOPCB4028849 E X H I B I T # STLCOPCB4028850 .^-$8&& 5V:- O . r> , v^ As a follow-up to-our phone conversatlon yesterday,' I sa . sending you copies-of :tfae lnfor-a&tlon on Aroclor 1260 study which was sent to.he by Dr. Kl*brough.'c Too say find this informative In preparing for your subsequent' discussion 'V': l When you have had a chance: to review your schedule, would you . , j kindly let ate know what days my be convenient for a meting. -Y :: I This probably will be in the Washington area and I anticipate 1 that you, Fcnate Kimbrough, Sob Squires and I will attend. - "'`v - - 2^`J- ;;.-:^-\..^;;sincnwiy, ,% > : .v ' . STLCOPCB4028851 HOT FOR PUlUCATIOa OR CIlCTJUnCK Aroclor 12-72-A AnlulJ Randoa brad fmli Sherman strain rata raarad undar specific pthogea-fre conditions vara obtainad fro* the SCDC (} flim at Lavreneeville, Ca. Aroclor 1260 Aroclor 1260 (Lot number AR-3) vaa supplied by Monsanto Industrial Chemicals Co., St. Louis, Mo. Statistics The students t-tast vas usad for comparing body vaights and veighe gain. Methods Four hundred 21-26 dry-old vaanling faaala rats, vaigbing 43-97 g, vara distributed into 2 groups of 200 animals each according to a table of randoa numbers. Each animal.vas vcighad and given an individual ear 1 tag number. The animals vara housed 10 rats par cage. Food and vater vers provided ad libitum. Two hundred animals vara fad the control diet of ground Purina laboratory chov; the test animals vere fed the 'same diet fortified vith 100 ppa Aroclor 1260. Aroclor 1260 vas incorporated into the diet by dissolving 5.2 g in ethyl ether then adding this to 100 g corn starch and allowing the ether to evaporate. The PO-cornstarch mixture vas blcndad vith 345 g ground chov, then pulverized in a mortar and diluted vith additional ground chov in a bakers sixer to give a 1000 ppa concentrate. The concentrate vas further diluted to obtain 54 kg of 100 ppa diet. The diet vas prepared every 10-14 days. Randoa samples froa the final sixes along vith samples of control chov vere taken at regular intervals to determine PCS levels in the control chov and to verify the 100 ppa fortification lavel. Ground laboratory chov vas also checked at Intervals for aflocoxin at the district of fics of the Food and Drug Administration in Hev Orleans. The ccabined body velght of rets in each cage ves recorded veeicly until the rats vers 6 months old, bi-veekly until 12 months old, and monthly thartsftar. Individual weights vert recorded only at the onset of the experiment and at death or sacrifice. The animals vere observed SCM 02281: nnncp'A STLCOPCB4028852 I1 Aroclor 12-72*4 - Fags 2 briefly each day tad animals exhibiting debilitating signs or largo tumors were removed from tha group cage tod housed individually. AC sach vaighing tha animals vara examined individually and abnormalities nocad. Food consumption vaa aaasurad on all rata during tha first two weeks of tha experiment, than during vaaka 5, 3, 11 and 20 and avary 12 vaaka thereafter. Autopsies vara performed on all rata that dlad. kata that vara aaerlficad vara killad uadar athar anaathaala by aera ring tha vana cava. Tiaauaa vara flzad la buffered foraalin and atainad with haaatozylin and aoaia. Additional special stains vill ba conduct ad. Zasulta During tha coursa of tha study 4 animals in tha taat group and 8 i controls vara sacrificed prior to tha final kill. These aniaala bad either large tumors vhich had ulcerated or hindered movement, or tha &n<mjal appeared moribund. In all, 14 aniaala la tha cast group died; 17 la tha control group died. One anlaal.vas accidently killed in aach group early in the study. Ho definite dose related signs of tozicity vers observed in the taat animals throughout the study. Cooperative curves of body weight gain and food consumption on the bssis of body weight, as veil as PCI intake of the teat group are given in Figure 1. A slight decline in the rate of weight gain of the taat group colored to the control group began about 3 months after onset of tha experiment. Keen final body weights ware 420 g (S.D. 72, S.Z. 3.4) for tha control group and 392 g (S.D. 62, S.Z. 4,6) in tha test group; tha difference was statistically significant (p<0.001). Food consumption (gm/rat/dsy) was comparable in both groups throughout the study. Kean weight gain vaa 330 g (S.D. 70, S.Z. 3.3) and 323 g (S.D. 60, S.Z. 4.3) for the control and test.groups, respectively; the difference in weight gain vaa also statistically i significant (p<0.001). PCI intake declined from 11.6 mg/kg/day during t tha first veek of exposure to 6.1 ag/kg/day at 3 months of exposure and to 4.3 mgAg/dsy aC 20 asnehs. A 6 g assn weight loss occurred in i both control teat groups during the 6 vaaka prior to tha final kill. SCH 022811' oooc STLCOPCB4028853 ?T*lisinary Hat of Micro#copic Finding# (Il*brou*h) !lui Hota: Section* of All Tiaaua* LLacad Vara Hoc Aval labia for All Animal* Tumor Typ - Mammary Gland: Control# (Tlaaua of ona rat autolyxad) Experimental Xata Fibroadanoma 13 14 . Adenocarcinoma 7 0 Pituitary: - Chroaophob Adenoma 38 Carcinoma Pituitary 0 > Utarua: 30 1 * r 1,1 Endometrial Polyp Endomatrial Sarcoma 20 2 23 3 and 1 adanocarcisoaa Thyroid: .- Medullary Call Tumors (parafollicular) . 26 IS I Livar: Nodular Syparplaaia 3 (8188,8192,1232) 7 . Nodular Byparplaaia and Hapatoaaa 1 (8231) 134 Nodular Hyperplasia. Hepatomas, and Hapatocallu- lar carcinoma# 0 Adenofihroaia 0 14 4 t*- t -2, --------. Bladdar: Papilloma 1 (8317) 0 In addition, a number of othar occasional tuaora vara found. SCM 0 2 2 8'; 000C35 STLCOPCB4028854 Pralisrinary List of Microscopic Findings - Page 2 Out of this group, 4 control rats and 4 rats in tht experimental group wars sacrlfiead early because they vara sick. All tumors of those aninsla, outside of the liver lesions, are included in the proceeding list. The liver lesions need to be added. The aniaals are as followsi Controls Experimental 8194 8131 8121 8234 8331 8347 8413 8317 ' SCH 0223*6 n /1 n T' nr' STLCOPCB4028855 . s~\ s' Figure 1 BODY WEIGHT ,I co . rsj 4 rvi O * Tn vJ * C 'J O O O t1 Ii I I I f l i i i % i D) Food intake gm/kg /day SCH 000033 STLCOPCB4028857 Figure 2 FOOD INTAKE AND PCB CONSUMPTION 1 2 3 4 5 6 7 8 9 10 11 1213 1415 16 17 18 1920 21222324 Months AKXXCR 1260: HhUas at d9 Janaary 31, 1575 The purpose of this see ting was to renew sections of liver tissue frcxa Or. Xlsbrough's 2-year study in which female rats were fed 100 ppa of AROCLOR 1260. Ore. Slab rough and Squire had studied and classified the findings. Dr. Levitt, either a visiting fellow or a post doctoral trainee did not participate to any great extent. He was sentiooed as haring a good knowledge of liver. Dr. Ricnter Had viewed the slides fro our 2-year study in which rats of both sexes had been fed 100 ppm, 10 ppa or 1 ppo of aROCLOR 1260. Subsequently, he and Dr. Gordon reviewed sections of liver froa all animals In this study. Three conclusions can be drawn. 1. In our earlier study, the severity of liver lesions wan greater in fessrles than in sales. ?. To a large extent, substantially the Basse type of lealonr were observed In both studies except that the lesions seemed to be wore advanced in Klabrough's study. Although there was some variation In terminology, the findings were reasonably close. 3. There were definite liver adenocarcinomas In flabrough'* study. Dr. Richter expressed the view later that 2 ani mals in our study approached the. type of lesion Klnbrough had observed, but there was agreement by Drs. Gor don and Rlc.iter that Dr. Kimbrough's rats bad developed a lesion which they had not observed In our tsrllsr study with AR0CL08 1260. If. Drs. Gordon or Richter feel that I have not sunaariied this eeetlrg correctly, or If they desire to saend or expand my remarks, I invite then to let s<a know. /bkp 2/3/75 'George J. Levinskas "iX m I I i \ I T if STLCOPCB4028859 STLCOPCB4028860 - ^ -c'/tlsn i.frcBf I)r^Xi*bro(d^,i*S-reax'i '3 . , ^5*;:v. were ,fed;100 ^ppatof-ABXacail260 . ^iv'551 Dra.^Ki^r5il^';a^Slrt;ia4:'teil*<l. -,' ^' . findings;;^Dr.^toYitt^^lCxc^iv;Tisiting doctors! tralnea-didnot'lpartieipa "' - -5 ' nB*r."'Blcnter )^^e#ed';^;lide*^rTOa our .^yeajrstady" which rets of both. sexes.-b&dlbeen.fedlOO ppd^lOppa or ^ ;?-;N -Sr:. >sfet*-V "s^sripic. y. ov., . *^'-- *. Zn cor earlier'atodjri `the sererlty^ liter letleoe eao *-** ~ greater in feles,,Jtnah'-in. -; ;?, To a large extent,"'substantially the saae type-of lesions ' ... were observed in both stu<Ue* except.that..the;l*8lens~-J. . ' seeaed to be wore advanced-In Xlnbrough'e study.:'1 Although' ' there wassoee.variation-in torainology, -the fiedings^-; .' r j - 3. There were deflnitefliver. adenocarcinoaas in Cnbrough's _. . study. y Dr. Richter.expressed the .view,1ater. that 2 .ani--"- J\..? sals in our stadjr Approached the gtype.of lesion Ha^ c - ,, trough had observed,.but there .was agreement by;Ore. Oor- > don end Richter that Br.! Kiabroagh's rats had .developed *' STLCOPCB4028861 E X H I B I T / STLCOPCB4028862 9 TOXiCOWOC* (NVIflONHCNTAl, SOCWCtfl CH(MltTflY UMT sciences mcoicav sciences !}rvjiifoicd BIO - TEST Jji6<Hjayiue, %c. 1810 SRONTAGC ROAO NORTH8ROOK, ILLINOIS 60062 March 24, 1975 *. coot On George J. Levinakas, Ph. D. Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 63166 Dear Dr. Levinskas: . I am enclosing the trip reports prepared by Dr. Richter and myself regarding our meeting at N. C. I. on January 31, 1975 with Drs. Kimbrough and Squire on Aroclor 1260 (IBT No. 622*07298). ., . .* 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, D. E. Gordon, D. V. M., Ph. D Section Head, Pathology Dept. DEG:hb Enclosure i DEPOSITION |I EXHIBIT | /S' 1 SCM 023632 STLCOPCB4028863 February 2, 1975 Trip Report On January 31, 1975; Drs. Richter, Levinskaa and myself mat with Drs, Squire and Kimbrough at the National Cancer Institute in Bethesda, Maryland. The purpose of this meeting was to review the slides and data from a 23 month oral feeding study with Aroclor 1260 in rats that was con ducted by Dr. Kimbrough while at the E.P.A. In her study, 200 female rats (Sherman strain) were fed 100 ppm of Aroclor 1260 in the diet for 21 months, while 200 female rats of the same strain served as controls and were fed 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 study began, and were sacrificed after 23 months on the study. Therefore, there was a 2 month recovery period at the end of the study in which the test material was removed from the diet. The details of the ex perimental design, procedures and results that were forwarded to Dr. Lav- inakas 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 Branch 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 02363- 00QC13 * STLCOPCB4028864 Trip Report .2 be deleted and re-classiflad it carcinomas or neoplastic no dales. This classification system was based on a Liver Tumor Workshop that was held on Dec. 11-13, 1974 in Silver Springs, Maryland which was 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-Year Oral Feeding Study With Arocior 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 ware conducted in Dr. Kimbrough's study. The additional glides 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 call 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-Teat study was also very low. The incidence of spontaneous hyperplastic liver nodules reached 20% in the Fischer Rat . ooor;.;^ STLCOPCB4028865 Trip Report 3 and in toms of the other strains. 2, ) Sex difference. On the basis of our studies those of other in ' vestigators in rats and mice, it eras found that the incidence of hyper plastic and neoplastic liver lesions are much higher in females. The results of the liver tumor workshop are to be published in the near future and will include the participants. The views of this group, regarding classification of hyperplastic and neoplastic liver lesions is based on the patho genesis of such lesions in rats that have been induced by known carcinogens such as the nitrosoamlnea and 2. - acetylanrinoflorise (2-AAT). The adaption of this terminology and classification system by the scientific community will have widespread implications regarding assessment of safety of chemicals and drugs currently on test and those previously studied. Attached are Or. Richter's comments in regard to Dr. Kimbrough's findings. . Donovan E. Gordon, D. V. M., Pb. D. DipL, Am. College Vet. Path. STLCOPCB4028866 Dr. Donovan Gordon Industrial Bio-Test Laboratories February 3, 1975 I wish to summarize my observations of the lesions seen in Dr. Kimbrough's study of Aroclor 1260 and the comparison of diagnoses with Dr. 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 h*n theirs. For examples they vtould call some of my hepatomas carcinoma 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 Dr. Kimbrough are using a new and revised terminology for the categories and list them as follows: My Terminology (Classical Use) Focal Hypertrophy Nodular Hyperplasia Hepatoma Dr. Squire's (Revised Terminology) Cellular Alteration Nodular Neoplasia or Neoplastic Nodule Carcinoma Carcinoma 5.) However, the lesions in Dr. Kimbrough's study were more severe than those in the Bio-Test study. The lesions that she and Dr. Squire are calling carcinoma are also carcinomas by my criteria. I would STLCOPCB4028867 conclude from an examination of their material that Or. Kimbrough's study demonstrated carcinogenicity. Ct/oAcf WardR. Richter. D.V.M. Dipl. Am. College Vet. Path. SCM 023637 000047^ STLCOPCB4028868 xxm I B I T STLCOPCB4028869 Way 25, 1970 Otis E. Fancher, Fh.D. Director Industrial Bio-Teat Laboratories, Inc. 1810 Frontage Road Northbrook, Illinois 60062 Dear Otis: * This letter will authorize you to proceed with a study with the leghorn chickens with the Aroclcr 1242, lot number AL-55* Shis should duplicate the study you did previously with Aroclor 1242, lob number AK-255* except in this instance you have suggested using dietary levels of 2, 4, and 8 ppm. 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. We 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 in St. Louis will more care fully characterize the two samples in terms of possi ble minute amounts of contaminants. Best personal regards. Sincerely, X?W:Ju User P. Wheeler Manager, Environmental Health STLCOPCB4028870 STLCOPCB4028871 TQSICOLQ4Y CMVIftOMMCttTAl SCJSMCSS CMCMISTVT . PUT SC1IMCSB MC040UI 8C13&C&B - fC *B - ie*i~ BIO - TEST Jk^Hofate, Skc. iaiO PRONfAGC ROAO NORTHfiROOK, IUJNOI5 60062 AACA eooc SIS rtiottOMc m*aoso September 30, 1971 Dr. George J. Levinskas Manager, Product Evaluation Monsanto Company 800 N. Lindbergh Boulevard St. Louis, Missouri 63166 Dear George: Enclosed are the. correction pages for die three teratology studies with Aroclor 1242, Arodor 1254 and Aroclor 1260. I discussed the interpretation of the renal caudal ectopia, found with Aroclor 1254, with Dr. Keplinger. He is in agreement with the interpretation that this is probably not a specific teratogenic effect but a general indication of the toxicity of the material. la reviewing the results from the three generation study there appears to be a reduction in both 5 day and 21 day pup survival in the group fed die highest level (100 ppm) of Aroclor 1254. This was aot observed in the other Aroder groups or in groups fed 1 or 10 ppm, again suggesting that the level of Arodor had a general toxic effect. As X pointed out in our phone conversation, die small differences observed in sex ratios are not significant. A statement to that effect has been inserted. I you have further questions, contact me at any time. Sincerely yours, /U Paul L. Wright Section Head, Toxicology . deposition I EXHIBIT i sievjiosxm'l 91 PLWilam Enclosures QnQC'0- SCM 02 3 58** STLCOPCB4028872 E X H I B I T STLCOPCB4028873 April 28, 1972 Dr. Otis Fanchar 624 Mo Abrego Drive Green Talley, Arisons, 83614 Dear Otisi 2 aa ashamed that 2 have lieenlso Blow; la- reviewing the manuscripta and sending you our ecssaenta* 2 guess Sep's sending me the chronic rat and dominant lethal and rhieken study on the 18th of April finally atlifted me to move. First of all, X agree we should hold up the publi cation of the chicken work until the current study - Is completed* .* The only other comment X have of substance la that X wish we could wo** In several paragraphs reviewing the Treon work on vapors published la 1936 (see en closed xerox copy of reprint)* A logical place for a review of those data would be at the top of the ~ 3rd page of your review paper* before your paragraph . at the top which begins with ^During succeeding - years little Information was reported concerning . . . the toxlcblpgy of taejftansytil McLaughlin etad. Host of the review articles oa the FCB's have failed to indicate awareness of the Treon work* X believe this is because the Rlsebrougha, Feakmlls, eta*, ete* -- the environmentalists -- have not encountered the American Industrial Hygiene Association Quarterly in the review of the literature nor .is it likely that '. they ever heard of the AXHA* _ . - . . " In any case,' X think the work Is worthy of* review even though it was inhalation data particularly since Drinker's earlier work la *37 1 fre<ju*ntly motioned and. this. . again concerned Inhalatiomv * . u J, r' ' . i ' * V.-- - ' ... In the last paragraph of this same review'paper you . Qr*Qf\;0 *4' * /* r **V SCH 023589^ STLCOPCB4028874 Dr. Otis Fancher April 26, 1972 Pag# - 2 - refer to the perniasable ambient concentrations of PCB's" as suggested by Trinkar in 1939. Actually tbs ACOIS establish*! th Threshold Limit Values based oq Tr#ons work rather than the earlier Drinker work, Z believe we nay have arrived at our high dietary level of 100 pp* in the studies done at Bio-Test based on the Treon work rather than Drinker's. Geers# Levlnsksas is reviewing each of the papers ' to see if you. Sop and I have missed typo's and be asy well suggest sene grammatical changes tJch we will enter on our copy and send xerox's to you and Sep. . . Z hope to get copies of all of the studies in the hands of Bill Papageorge, Scott Tucker and the lawyers next * week. Z do not anticipate a lot of changes from the* and hope that the attorneys agree that we can go ahead with publication. Best personal regards. Sincerely, S?*/bks cos if. L. Sepllnger User P, kheeler Ifcr., Environmental Health nodical Department ' onQCVt _ SCM 023590 STLCOPCB4028875 u jx x I I \ B I T STLCOPCB4028876 *o*tceio* CNV1 AOXM CMTA4. SCltMCCS CMCMISTQT ' 01AMT SCJCMCtS m COICJU. teiCMCCJ BIO -TEST J^zlyyiGjby)jigt Stic. 1810 RRQNTACS RCAO NORTHBROOK.UUNOiS 60062 April 13, 1975 ^ 2 i A*CA C33C JU tcwxp**o.nc jti- iC2 Dr. Georg* Roush., Jr. Monsanto Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear George: I fully appreciate that the meeting on PC3*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 next 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 [1 DEPOSITION | EXHIBIT IjLEViiOSyaVS-ffi STLCOPCB4028877 \ E X H l B I T / STLCOPCB4028878 1 Jt !- Dr. Georgs Urbuku . ... . . .. U*ou1d < . ,*. . jMctiaa of U^V ' 11BT l^ibasfVscaattf rtf-siamlasd as > tUtt TH (lOppa )mtt ;- Sst iiraD tbs ortiiul block of ambeddod tlaaes v'..yjte Jspttic^lsslo was srlgiaally classified by Or." Richter ka '& ip^iotei^x*rimw,Dr. 'Wetter has classified tkla lellM ii wxhUr hyperplasia ^ad 1 eoacg. However, yos viU alas note that whan additional soctia&a of Urar (rom this animal were processed CBT Study Mombsgjsd)-06672), W fiver alteration* van s'! srwad which attests"is . -- haters f Uwm Ja(aatiooaass*^?L. '; -ajjrS.- t'^Ji^TlUa chugsya ce-c report of IBTMa^ttfrOM rvUato.%; ... ' ... .* ' _ iffiiria or rsirissi jf7/; aactesW a ailds of tbs absTs'.-^r. JKacsrsly,. ..V , - Jr._ _ -. * '/ V:/ - - / c. * ` ''` Gordon, D. V. W. Pb.D. Sactioa llwf. Pathology DapartrooBl sac losers. i i f I < 4 i: ooonca 4-L STLCOPCB4028879 >. V/.'A.v ' 8S2e#'rV .0 *- ;- ^ ~0Z*r:^' - * ' r''-\'-ir/- : ,^ V J '7'-* > " s* "***>*fBc.iiitAotyoffcctaoiAdhatoAifewaiAAfcii^atoih2<lhlto*tfA40tta*furM- ,W;VAA&*nh1^^a>Mt? rAiy4JiaTi .-, -'- *''" ' A&OCu3ir- 1249|j|'" Tabulation f ladlrtdoai Unr LaaWax in Jtala ^ 154 L^-iy ^ Sacrlilco Lata ml 24 Month Doi Laral \aima No. and Sax . * o i I ii M Is ia.f* gS 1* os u c-m 740M lOOppm 722" 724" 744" 744" 744" 7441" 771" 4 775" Ext" + 44 733" 745" * 766" 4+ 767" 776" 773" 786" +44 789" 793" K444 Ext" 44 Ur*r L*loa a IS II HII 11 St i z S it = 8. f o4y Sto P P P P P vto * = !i 1 1 2: 0 C40 O U 3 to 12 f 6 3U * e ii --: es Control 8M 1!" 12" 21"' 23" 33" 34" 35" Ext" i 46F [ 47" j 49" 51" 52" 56" 59" 62" 4 STLCOPCB4028880 i E X H I B I T # STLCOPCB4028881 Monsanto Monsanto ComoJftT 800 N Lmdborgn 8culv|fd St. louit. Utioun 83163 phona: '3u: 394-1000 July 18, 1975 Dr. J.C. Calandra Industrial 310-TEST Laboratories 1310 Frontage Rd. Northbrook, 111. 60062 re: ARCCLOR 2-year Rat Feeding Studies Dear Joe: The attached table summarizes a comparison of the 3 revised. AROCLOR reports (1242, 1254, 1250). ' In 2 instances, the previous conclusion of "slightly tumcrigenic" 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 1242 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 1242. This leads me to conclude that the numbering scheme shown in the second set of reports is correct. With AROCLOR 1254 confusion is compounded. The original report showed tumors in animals with numbers in the units to teens, out 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 STLCOPCB4028882 Dr. J.C. Calandra July 18, 1975 Page - 2 would be supplied. This has not been done. It may not be necessary for APOCLORS 1260 and 1242, but APOCLOR 1254 remains unresolved. Insofar as I can see, the remainder of the reports appear acceptable. Kindest personal regards. Sincerely, George J. Levinskas, PhD Mgr., Environmental Assessment and Toxicology /bkp att. cc: Dr. George Roush, Jr., M.D. G^QCCC- tf'"' O-f A A a STLCOPCB4028883 Product Supplemental Report #1 (mailed) AROCLOR1260 . conclusion hepatomas. . slightly tumorigenic r* 3 range of test animal nos; p. 9 ' 600 to 800 p. 10 1000 series p. 11 70 to 100 p. 12 500 to 600 p. 13 600 to 700 p. 14 700 series Supplemental Report #2 (JCC 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 tumorlgenlc S slightly tumorigenic S' AROCLOR 1242 conclusion . slightly tumorigenic does not appear carcinogenic hepatomas range of test animal nos. 7 3 as in report #2 as in report #1 for AROCLOR 12(SO AROCLOR 1260 SCM 022804 " onoc: STLCOPCB4028884 xxm I B I T # STLCOPCB4028885 /J lice L. f ji] ( roiieoiair KNV'VQ.MKMTAC ICICNCU CMCH'ITIT C*KT lOCDCtl mcoic-ai. (cicmccs SfiSLuJdAicJ BIO -TEST -Pfilwtfihy/\i0 Sue. 1810 P RONTAG8 RQAO NORTHBROOK. ILLINOIS 60062 August 4, 1975 A DCA C30C 3>* TtUCPxONt 171-303 ( Dr. George J. Levinskas, Manager Environmental Assessment and Toxicology Monsanto Company 800 North Lindbergh Boulevard St. Louis, 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. Sincerely yours, ^POSITION r EXHIBIT JCC:AR J. C. Calandra President SCM 023620 STLCOPCB4028886 E X H I B I T # STLCOPCB4028887 16 a. Summary - (_ . In moat Instance*, the spectrum of treatment-related Histopathological . findings In the Uver from this re-avsIuation dSd net 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 (104 ppm) of the 24-Month Sacrifice, Wo hepatocellular . - carcinomas were observed. One Uver turner (a hepatoma) previously reported ' *-* -v... '"*,v 1 < 'tyvr ' ' ' ? * ' ' in a T-tX animal No:r445) -was rv'classified as nodular hvperpliila- The * other <T*atoeat-related "lesions reported are Tygarded'aa degenertave or'.'; - . V *. "V; ' \ \ ' ~ . *--< r t'' ` . hyperplastic in nature arid they ire morphologic manifestations ot sn scapd-e ~ * - - u' ' ` > . " . .. response of the liver ascribed to biatr jns/ornabon of the test material. The 1 * f , f. latter lesions were confined primarily to test animals of the 12 and 24 month v.- sacrifices and they were dose-related la taddenes and severity. ' ,, '. h>"e..V >S't.-*- -s'* . * . . -- - ,. In conclusion. Aroclcr 1254 does not appear to be carcinogenic in rate ' : ->t ... V ; ~JbT. " ' :.V. W* <- i'- ' fed tor two years at levels up and including 100 ppm- i'-*''r~ ' ' . ' . 4 .. . j, .' Respectfully submitted, ^ *' . . * .' ./**-' -uj-;. . INDUSTRIAL 310-TEST LABORATORIES. INC. ,, - . Report Prepared and Rsviswed byt ; .- . -v/' . x` D. S. Gordon. D.V.hL.Pb.D. Section Head. Pathology . " .. Report Approved byt .. - . " *; */* ' ..'A; v Ttf' - . .L* .ir.--- --A .-C-iaii .. id. X~ ^ICsplUfg*Jt. 0. -:*.Tlp. . -- M'" nan*nVflw**-rTe-&,Vr r*-, 'J' -g-r-V^.-. .* J. jf. `Jr-.. --*V*,w' .. . ,, ... -' - w sM deposition i EXHIBIT vI .,, . Lgy/ioS^i 53 !>/sV Vev'v-' 4y-'-'xV oooesa^ STLCOPCB4028888 MJXX I V I B I T STLCOPCB4028889 2nuituzl 810- TiST JMctatcnte. Jha. ~ 1910 MONUQe ROAO NORTHBROOK, ILLINOIS *0*42 August 5, 1975 172 Georgs J. Levinskas, Ph.D. Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 83166 Dear Mr. Levinskas: Re: IBT No. 641-06672 - Histopathological Evaluation oi Additional Liver Sections from Rats ol a Two - Year Chronic Oral Toxicity Study of Aroclor 12S<- Enclosed pleass 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, 4s JCC/fd J. C. CnUndra President ' Z DEPOSITION | EXHIBIT 1 -11 u^c .. nvwwerffpyyyy. t y '..*[-**.*.--***-*( STLCOPCB4028890 V E X Hi I B I T f* STLCOPCB4028891 t August 14, 1975 Dr, J.C, Calandra Industrial BI0-TS3? Laborstorits 1810 Frontage Road Sorthbrook, HI, 6006Z re* ABOCLOB 2-year Bat feeding Studies Dear Joe: With respect to your letter of August 4, 1975, * appear to be reading froa different scripts, . The attached copies of pages froa the revised ABCCLCB 1254 report (XBT So, 641-06672), dated March 24, 1975 shoes -that virtually all anlaal numbers have 3-digits. The exceptions include 4-digit number on page 10, a series of 2-dlgit numbers ranging from 71-70 on page 11, and slides marked Srt or Sx'appe&rlng c pages 12 and 14, Copies of pages 83-88 froa the original report, XBT 2fe. B7298 dated Boveaber 12, 1971 also are attached. These contain the tables listing tumors, and they are the only ones in which Individual animal numbers appear. As summarised below, there is a repetition of low digit numbers in each group. Mel rat nos. Female rat nos. Control 1,2 46-43,50, 52,|9,64, ABOCLOk 1254 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, Hera- * DEPOSITION i EXHIBIT I 5-2W 1A6 sc* 02280 00QCG1 STLCOPCB4028892 Dr. J.C. Calandra August 14, 1975 Page - 2 - ever, even in thos instances where tha same numbers appear, there ar inconsistencies as shown blowj - ' group and sex animal no. . P*g *1 -?1, 46 ' 1 ppa Female all shown as 47 1 ppa Female control 4a 1 ppa Female females 50 1 ppa Female 52 10 ppa Mai 10 ppa Fecal 100 ppa Male 66 100 ppa Female Although 'It is of minimal significants, pages 84 and 85 of the original report aro mlsnuabared in that pag 84 is a continuation of th table which begins on page 85. Tha paga for the AEOCLOE 1254 containing ths phrsss "does not appear_to b carcinogenic0 has bn received. V not the interchange of tha numbers 1242 and 1260 on thos 2 AEOCLOH reports, end sill destroy ths mislabeled copies. Sincerely, /b*P att. Georg J0 Levinskaa, HID Mgr. 9 Environmental Assessment and Toxicology 0Z28a sc* G0G0G2 STLCOPCB4028893 STLCOPCB4028894 I To*ico.oaT * cNvtnoNMcNTAb icuMcts cwtMiarnT ukt sciences medical sciences !)rt&u2>tAJxd BIO T E S T Jjxky&jjywsA, Sac. 1810 f PONTAGE ROAO . NORTH BROOK. ILUNOIS 60062 October 17. 1975 ' - u coot 111 TCtCA-ONC Ut-JOJO George J. Levinskas, Ph. D. Mgr., Environmental Aaaeaament k Toxicology Monsanto Company 800 N. Lindbergh Blvd. St. Louie, Missouri 63166 Dear Dr. Levinakaa: 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 ;>*c/zr Donovan E. Gordon, D. V. M. , Ph. D. Section Head, Pathology 2 DEPOSITION J EXHIBIT i N'>')-?7 tP/e- SCrt 022-82' 000CC3 STLCOPCB4028895 m xx t i B 1 T # 1 4 STLCOPCB4028896 INTER-OFFICE CORRESPONDENCE JUfcriW 8IO-T8ST Ja MR Dcaoikr 24( 1175 ro PCB Papers George Lcvinskas has some questions regarding the pathology section of the PCB papers. He wants to visit Northbrook after Jan. I at a time convenient to you. Please make the necessary arrangements. CLK/Um G<. o. 7Ls Kennedy ^ '*a) v0^ \ooo e^` ,, - ooocg-v STLCOPCB4028897 UJXX I I B I T # J STLCOPCB4028898 Dr. LeyinatuTa wtU habere on.Friday', Jamury 1, 19T6 to dtacoss ' X/'" ,Ji :&.&*--V-. ,5;V ;. Sg'.-iyf- f.jf'. " -'-;k L- anptton of the P^CD pape.rs.-V-"- tha pathology section of the PCD ';U 3V r. '-\J ' f?\ . 1.* f' .` 1^>` ' .. 5. - ;'V. r%>s* . January 12 1996 iv y ... STLCOPCB4028899 V E X H I B I T # STLCOPCB4028900 ) Of Toxlclty and Environmental Effects of Commercial PCBs V A<v- f. 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/m3 and 0.5 mg/m3 have been established for materials containing average chlorine values of 42# and 54#, respectively, of the available sites. 3) The no-effect level for these materials in chronic t DEPOSITION ( EXHIBIT 1 S'Snsi oJic SCM 019639 nnnriCo STLCOPCB4028901 .0 ) rat and dog feeding studies is about 10 ppnx 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 teiatclogy 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 0196^0 G00CG7 * STLCOPCB4028902 ) groups except males fed AKOCLOR 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 silch 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 AROCLORS 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 AROCLORS 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 ' ' GOQCGSF- STLCOPCB4028903 (> ) rats and mice. In the chicken toxicity/reproduction study, AROCLOR * 1260 was without effect at all test levels. AROCLORS 1242 and 1254 at 100 ppm in the diet decreased egg hatchability. In fact, poor hatchability 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 a. dietary level of 10 ppm. Biodegradation studies were conducted using semi-continuous activated sludge systems and tissues of rats fed FCBs 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-evaluation 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 SCM 0196^2 00GGC0 ^ STLCOPCB4028904 o) 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 ugAg/day. It is higher than the safe human intake (1 jjgAg/day) estimated by FDA from, human data when it promulgated its tolerances for PCBs in food. O_ x SCM 0196^3 00QGr 0 STLCOPCB4028905 o) Human data (F.R. July 6, 1973. p. 18097, Section (2)) Lowest level of PCBs producing effect In man 500 mg total. 500 mg consumed by 50 Kg individual over 50 days. . 500 mg 4. 50 kg 4. 50 days * 200 ufiAg/day (effect level) . Using 10-fold safety factor leads to estimate that 20 ugAg/ day may ba-safe over a 50-day interval. Since PCBs have long half-life; calculating same intake over & 22-month span arrives at an estimated safe intake for a pro tracted period of 1 ugAs/day. . Primate data (Fed. Proc. 24:338 (1975). Abstract 675. Barsotti & Allen) At 2.5 ppm in diet, primates ingested 182 mg over 52 weeks. 182 mg z. 385 days =0.5 mg/day which produced effects. Assuming 5 kg primate, 500 yg -f 5 kg 100 ugAg/day Conclusions: Primate study done at dosages (100 pgAg/day) greater than the safe dosage (1 ugAg/day) esimated from human data. Primate study does show effects at lower dosages over longer time span (100 uSAg/365 days) than had been observed in man ' (200 ugAg/50 days). The too<*2 amount of PCBs producing effects in primates (l89 mg) was lower than that producing effects in man (500 mg). (N.B. Abrahamson & Allen, Env. Health Perspectives. June, 1973, 81-86. Report that infant monkeys are aole "to tolerate doses of PCBs that produce extreme morbidity in adult monkeys.) Dog and Rodent Data ' (P.R. July 6, 1973. p. 18097, Section (1)) "No-effect" level for man using a 100-fold safety factor and accepting 10 ppm as a "no-effect" level in animal3 would be 2.5 jig/kg/day based on dog data and. 3 uSAs/Uay based on rat data. \ ' If rat data "no-effect" level drops to 1 ppm, then corres ponding estimate of human "no-effect" level drops to 0.3 \xs/ 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^^ STLCOPCB4028906 PCB Primate and Human Residue Data Cone. In diet, ppm Intake, ugAg/day Estimated safe dose, UgAg/day Liver conc.,ug/g Fat cone., ug/g Primates 2.5 100 56.3fo.oi)* 50.0(27.7)* . Walue in ( ) from infant primate. Allen et al., TAP 30: 440-451 (1974) **Tobs, Env. Health Persp.,79-81, April 1972 Human -/ - 0 (314 samples)** <1 (125 samples) 1-2 (I65 samples) >2 (33 samples) scm 00QC72 STLCOPCB4028907 xxm I \ B I T STLCOPCB4028908 oncnvMT *eic*cta MKOICAW kCKkCCA , . ; f ' S Goorpe J. Levin^kaa, J'h. D. s4- -f * Monsanto Company ' 800 N, Lindbergh iUvd. 7;4 - - a' - . ~ -W> v-TV. 0St. Louis, Missouri Oil66 . . '"r. '. Dear Dr, Levinskas - t* ^ > - . \.*.'?**!'. e\i- . '**. ' v&*v; . . .**:.' ." ' * . ' -" -'; ' I am enclosing tlw revised''luriwr tables for IheCAroclor paper - that we discussed during your >isit on January 15, 1976** ,You will . notice there arc two tables with the saute data, * One table is a listing <W tumors by organ systout, as you suggested, to replace' the original table* { think I also prefer (hr former although it will require -moro space when printed, !: ' ` - >,*. *... *. - * -'* ' . VjOj^r* 'f. fm ... ' r~J '* "jf ' ; ' Should ywu have any further questions, please contact me, , : \- -| . ' *' " ,' '***- *. * *^ - Sincerely yours* - ' . * ` . . v. ^ ;/ "ir. ' * 'i * v" . ' . ;/ 7 * -/-/ .-**%* . ?'tkftflvnn E, Cordon,' O, V. XI., Ph. D . . Section Hoad Pathology . DEG/ji . cc: M, L, Kcplinger ^ .u . ' ` *3 *. '* i.i --h*r.^va/r-L .-C-V* . '. ` ^ DEPOSITION | EXHIBIT |L6V/dstfls-3c mm onnr,7'T STLCOPCB4028909 STLCOPCB4028910 m Monsanto D*pt. jf Medicine & Environmental Health- 0 J. Levinskas, A2SA l L < November 17, 1978 AROCLOR 1260 *4 > |it*CI 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 t DEPOSITION | EXHIBIT flSv(,i*AS'3l On Wednesday, November 15, 197 8, I had a call from Joan * Ranson, an industrial hygienist with OSHA in Philadelphia? 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 haul 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-IOM (HEV. 2/78) STLCOPCB4028911 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 / Geoi _ Levinskas SCM 019638 STLCOPCB4028912 U1XX V \ I B I T # STLCOPCB4028913 Monsanto rmom .**( * tOCifON. 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 ail 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^ STLCOPCB4028914 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 cf PCBs. In some studies, the test material has csen 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 ^sseL' 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 were 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 _ - . . " SCH 0197L7 ' goqc/t? ^ STLCOPCB4028915 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 Monsanto1s 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 AROCLOR 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 019718 " . ooocvd"1 STLCOPCB4028916 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 'concluded that the continued use of AROCLORS^ as dielectric fluids will not present an unreasonable human health hazard. SCM 019719 GOOC70 STLCOPCB4028917 bcc V. B. paoageorge - B2SX } d^ - January l&j 1975 Dr. Donovan 3. Gordon Industrial HO-TSST laboratories, Inc, 1810 Frontage Hoed Borthbroo*, Illinois $006* Bss Polychlorinated Biphenyl (103) Dear Dcot V '* - . *'/*-.* . Inclosed are copies of three reprints dealing with liver tuaorlgeniaia of FCB's. these are froai . (1) Gann. (1972).,2,1 805. (2) Sana. (1973).*i 105-106. -- - (3) J. gatl Cane. Inst.. (1973).21* 1637-16*6. At studies were perferased with Japanese aatarials (Kane- chlors), but^ih*-eaatj are of direct Interest to our seating oty^OCLCR V2S0y Toe and Sr. Mehtar should be faaill&r uTth-the contents of those papers as you review the ABOCLCR slides. ' Our Meting Is scheduled for January 31# 1975 In Kooa 201, Building 37# at 0GX in Bethssda. e elU be Mating with Srs. Hah rough and Squires. 80 specific time has been set, but we can plan on starting In the boruing. I will confir* a specific tiae and will check with you or lap oa a place to stay the night before. Sincerely, *1* ec Dr. M. .. Xapllnger George J. Levinakaa, Ph. S. Manager, Invironmental Aasesa- aent and toxicology * DEPQSIT10N | EXHIBIT SLEVipj^Cr <^. /3 IS'in.-gn ujjt SCM 022820 OOOOIO ^ STLCOPCB4028918 L E X H I B I T # STLCOPCB4028919 TO c 7 73Air. T. H. Bottini - B2CA ^ Bi. P'.--T "DuGaimu B3NA ^-Dr. W. C. Hammann - B3SA`^1I#,1 Air. E. H. Harbison - DlK Dr--Gt--I XMr. W.- B. Air. D. Wood - B2SD-2.0- t~ s Abe. C. Paeon - B2SC Ate.R. G. Potter - B2SB i""! Dr. G. Roush- A2'SA Ate. J. C. Weber - BIND `1 T Air. W. W. Withers - B2SA-2''cr/ <Pr**y??;-rtfrgfat:: - 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. Yoi\r cooperation is appreciated. M-fi- /mka acta fhmentj D. R. Bishop fa tL^. '{A * deposition exhibit `-svtNjt/vs-c SCM 058053 IH tg XCV. t 000C30 STLCOPCB4028920 V E X H I B I T # STLCOPCB4028921 -------------- ------ - D. R. Bishop - BIND o* f i Nov. 17, 1975 (".(.Cl TO Mr. T. H. Bottini - B2GA 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- /inks attachment * D. R. Bishop IN 11 "W* 1 * - deposition 11 exhibit cot SCM 058337 * QnQCGl^i STLCOPCB4028922 . . .. IMMEDIATELY 1975 " . MONSANTO INDUSTRIAL CHEMICALS CO. ' D. R. Bishop (314) 694-2891 PUBLIC RELATIONS DEPARTMENT 800 N. Lindbergh 8oulevard SL Louis, Missouri 63168 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 (PCB) 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 Ctnaha. Aroclor is a Monsanto trademark for a class of chlorinatec 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 058338 STLCOPCB4028923 --2 During his re-evaluation, Dr. Pour saw the same alteratic 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, althoug! 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 Qnoc.SQ: STLCOPCB4028924 --3 opinion, difficult and sometimes impossible to distinguish between regeneration, hyperplasia and neoplasia, particularly when the tissue is continuously exposed to a toxic substance.1' 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. 63 SCM 058340 onOCX* STLCOPCB4028925 STLCOPCB4028926 Sk&ujJaLgJ BIO -TEST 1810 FRONTAGE ROAD ` raxicoLOOr ch cm i ir*r MCSICAL SCIENCES Micoio^oor |"OUTiaw *a#cty 4 MCAUTM NORTHBROOK,ILLINOIS 80082 July 31, 1981 SVic. AACA COSC 3t2 TCLA**OHC 271-3030 Tcur* 72-A^ar ` Dr. George Levinskas Monsanto Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear Dr. Levinskas: As per your request of July 30, 1981, Dr. D. 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, MAB.-AR cc - Mr. A. Uelner Monsanto Company Marilyn A* Biederer Validation Assistance Specialist SCM 023698 Qn STLCOPCB4028927 E X H B I T # STLCOPCB4028928 Monsanto De, Jf Medicine & Environmental Health ,J, Levinskas, G2WF (4-83C July 9, 1981 AROCLOR Products 1242, 10e;A and 1260: Two Year Rat Feeding St J.R.G. Ortiz, E2NU A.F. Uelner, G2WC T0 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 1N-1QM (REV. 2/73) SCM 05S9Z9 STLCOPCB4028929 wxx \ I B 1 T STLCOPCB4028930 ' Monsanto C- Sjitk of Nominee Lo<j;-on Paul L. Wright Co^oany unit Of Staff QOt. Oiv.tion 6ut.nt Group QOartmnt Creve Coe1. Corporate Staff Pot.{.on T,u Medical Salary Grad* Annual Salary Medical Oat* of Lart incrat* Toxicology Manage: Ptfiof manci Excellent 19 $28,980.00 Gr o wtn Excellent 2/75 Oat* t_itl Cmoioy* 2/75 Tyoa of Awird (Select tne award category from tne opoont* Kd of tn.t meet wfi<n motf aoeroortatary o**crie> tne award and enter m category *o<a t coot oelow.) Staff ____________________ Awir0 Category Category Code Nunc Solution to product toxicity problem which permitted Monsanto 28 cxtcrio* tn* icni<mni jna <u mmficinM to Monunto Biowto continue product manufacturing 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 r 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 Food and Drug Administration (FDA) at 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 oy 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 Phe 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 containing maleic anhydride in food contact applications. Thus, we relieve that Dr. Wright played a singular, outstanding role in preventing 'DA from publicly proclaiming that maleic anhydride was a suspect. STLCOPCB4028931 it a i .j- V. - - carcinogen. Because of the many uses of maleic anhydride in foci 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 053800 STLCOPCB4028932 U1XX I I B I T STLCOPCB4028933 Monsanto C- .Hie Vs MS \ N,m* of Notninw T,. Wright Comoany Unit or Sta<* Oapt. Oivtiton Med. FTnv. Hlth. Poiihon TU . Salary Grade Tn*icoloav Manager 19 Par f ormanca Sutineii Group Annual Salary $31,800 Growtft Department Location Creve Cceu; Oata of Last Ihowm 2/1/76 Oat* L*U Employ* R*vi*w Typ of Award . (Saiact tha aware catayory from tn oppotit tida of tht* tnt wn*cfl most approprutery dascridei tft award and antar tn< category an< coda da tow.) . Award Category Technical ^ Accomplishment of significant results 0*cr>B* tn icnl*rmnt and it* tiynifieanea to Monunto otowi Category Coda Ni 106 At a recent meeting in Creve Coeur, Glenn Schweitzer, head of the Office c 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 si 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 polychlorinate 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. Racommenclad Byt (Rv. 10/7*1 Oat Award Amount R commanded! Approved Byt N--y Oat Award amour 'approved to payment* iMl. 7?/W r SCM 058795 0005.05- STLCOPCB4028934 U IX I I \ I B .5? | l STLCOPCB4028935 , wua i i> t AUTHORIZATION TO RELEASE INFORMATION (PRIVATE PERSON OR ORGANIZATION) TO PROBATION OFFICER TO WHOM IT MAY CONCERN: It----------------- Paul L. Wright, the undersigned, hereby authorize the United States Probation Office for the EasternDistrict nf His sour i or its authorized representative(s) or employee(s), bearing this release or copy thereof, to obtain anv information in your files pertaining to my: [jf Employment 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 anv 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. STLCOPCB4028936 P(3/?$048) UO uHr*D sr*rgj diw^c: couwr mocral eao8a rion jtitim REQUEST FOR EMPLOYMENT DATA AOORESS 0 P PROBATION OFFICE 111 U.S. Court h Custom House 1114 Market Streec St. Louis, MO 63101-2071 r - Personnel Department Monsanto Company 800 North Lindbergh Blvd. St. Louis, MO 63141 L 3/7/Si Oear Sir: The person identified below is under investigation by this office. The information requested is needed to complete this investigation. Your cooperation will be greatly appreciated. Ptesse return this form Tt,k`n days m the enclosed yr^titde f| necessary. Patricia/VJ P/ds 1 N*e o pejnoS^tiwj'uJ'vtsTioATto r.*< WRIGHT.. Paul Leer, tooncss or person scjmc investigated 2001 North Geyer Rd. J St. Louis, MO 63131 States employed trom Nov.,1972, to Feb. 29, 1984, as manager of special projects in Dept, of Medical & Environmental Health earning S55,QO0 per year. Please information desired verify. EMPLOY? _WAS THIS PCJRSOM CVCA IM YOU* ________C w IP Yli*' CIV* OATS STAMT.CO /////72- OAT* LEFT SAynr on wacc POSITIONS HCLO REASON POP TERMINATING '("'LoT","T^y-r- '^-<7 / WAS THIS PERSON'S SAUART *TT*ChCO' Y*S wQUCO YOU COMSlOCR RECMPi_OYtNC THIS PCRSONf Cj Y remarks ('C<***n4 (Ala pwrvtfi'j atrondanca, aMJifr* Jnetapor, ra/J*Jilfr. nrf aitfan-t rta*aa fpVaf ad *P rw* ar*a*faar*oRO AUTHORIZATION TO RELEASE CONFIDENTIAL INFORMATION form attached. i o,0::.O7 STLCOPCB4028937 E X H! Ii B! I STLCOPCB4028938 Toxicology Section/St. Louis (CQ-/OI V_/OEPT_/LOCATION) SPECIAL rnrPE OF REPORT) REPORT WO C<--M( CO H CM Csl XT CM O *~i 2 Q*-c--o* o_ai .C CJ < ft. 3a u. vt*oa; o to a. oi ^4 CI3 <n a > *C""C OIwj f.m-13 ><u uo w J oi s: < E- "J t) fot. oE-* box o ft H O O ' cm t) CJ rl o xa a2 f- < iii D < 6z oz > uccj 8a REPORT NO.: MSL-2002 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE- TOXICITY OF AROCLOR PRODUCTS 1242, 1254 AND 12 60 TO THE LIVER OF ALBINO-RATS authors: George J. Levinskas, 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. ^position' i j EXHIBIT i i ^iHHHi-401 LCkL SCM 058930 000/03 u STLCOPCB4028939 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. Levinskas, G2WF 9 - J.G. Nassi'f, E2ND '` 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ A8STRACT 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 recordsc^n be changed. goo:\o3 R- 1088(C) (Rev. 1/SOI . SCM 058931 - STLCOPCB4028940 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 the3 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 3 -1 SCH 058932 STLCOPCB4028941 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 . - sW0^ STLCOPCB4028942 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). SCM 05393** G3Q:* STLCOPCB4028943 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 feeding1 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 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. 'Tile uumbermg- sequence SCM 058935 000:'. 13 ^ STLCOPCB4028944 anri^-the..~4esicj.atiDa.< of . some. ..an imals ^aa.JlExtra."*. suggests that additional animals were, plac^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.8 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.8 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^ STLCOPCB4028945 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 ooo:'.:5^ STLCOPCB4028946 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 SCM 058938 cno STLCOPCB4028947 Footnotes 1. 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. 2. R.D. Kimbrough, personal communication. 3. 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. 4. Dr. David Ford of Bath University. . 5. 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. .` 6. 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. 7. Charles River Breeding Laboratories, North Wilmington, Massachusetts. 8. 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- ' ^ 0S. ooo:'.'/7 ^ . . #**-... STLCOPCB4028948 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 058940 nnfV ' STLCOPCB4028949 References 1. ACGIH (1969). Threshold Limit Values of Airborne Contaminants. American Conference of Governmental Industrial Hygienists. Cincinnati. 2. Caiandra, 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 Caiandra, J.C. (1972). Reproductive effects of polychlorinated biphenyls in white leghorn chickens. Toxicol. Appl. Pharmacol. 22, 316. 12. Keplinger, M.L., Fancher, O.E., Caiandra, J.C. (1971). Toxicologic studies with polychlorinated biphenyls. Toxicol. Appl. Pharmacol. 19, 402-403. - 10 - SCH 058941 nnn-' ' STLCOPCB4028950 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. ooo:*.-0 SCH 055942 ^ STLCOPCB4028951 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 Honths 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 Cholangiohepatoma 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 11 1 1 00 023 111 533 000 000 9 0 0 8 8 3 3b 1C STLCOPCB4028952 * 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". k Includes 1 animal without record of examination in or iginal IBT report. Not listed as hepatoma in worksheets for Gordon and Richter report but appears and1 wasi crossed out on typed tabulationi for animal marked "Extra". .^ for other 2 animals do not appear in records of examinations for original IBT' report Diagnoses O' . Diagnosis does not appear in records of examinations for original IBT report. Ul ( J r- > Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254 : Primary Liver Lesions Among Albino 1Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terrains 1 0 l 10 100 No. Aniaals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cholangiohepatoaa 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 26 179 131 120 034 103 563 000 000 13 1 1 14 14 4 4b 2C * Control group has animal dead at 19 aonths and 2 Barked "Extra". 1 ppa group has animals dead at 22, 22 aontbs, 1 marked "Extra" and 1 other for which date of death is not recorded. 10 ppm group has aaimals dead at 22, 22, 22 aonths. ,,100 ppa group has aniaals dead at 23, 23 aonths and 9 aarked "Extra". b Includes 2 aniaals marked "Extra". Diagnoses do not appear in records of examination for original IBT reports. STLCOPCB4028953 Oc 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. C n ! Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats Sacrifice Interval Diet Level (ppm) 3 Honths 0 1 10 100 6 Honths 0 1 10 100 12 Honths 0 1 10 100 Terminal* 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 Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytea Nodular hyperplasia Ductular hyperplasia Hepatoma Cholangiohepatoma 202 3 0 0 4C 0 000 0 000 2 000 0 00 1 0 000 0 000 0 1 12 6 1 0 ld 0 022 0 000 3 000 0 000 0 000 0 000 0 125 000 000 000 000 1 10 000 000 3 0 0 4 1 2 0 0 157 9 143 6 10 1 0 0 3 13 9 107 6 5 6 7 12 0 0 le 7f'* 000 48.h *.Control group has animal dead at 19 aontha and 2 marked "Extra". 1 ppm group has aniauils dead at 20, 22, 22 aumths. 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. b Includes 1 marked "Extra". c Worksheets show listings under this diagnosis with arrow pointing to Vacuolar change column. d,,Date of death of thia animal not recorded. * No Record of examination for original IBT report. Listed on worksheets for Gordon and Richter report with diagnosis crossed out. * 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. * Includes one animal with both diagnoses. Hepatoma is 1 of 2 crossed out (superscript f). b 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. ! xxm B I T # STLCOPCB4028955 jruhuitial 8 I O * T E S T Jkh. -' ` ' . REPORT TO ` MONSANTO COMPANY . TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH ARCCLOR 1254 - IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL UVER SECTIONS 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 1254 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. . i DEPOSITION | j EXHIBIT I /1 SCM 058166 qoq:~i ^ STLCOPCB4028956 9ntLui*lal B I O - T E S T JsJuyvaiytud. jke. 2 II. Summary In moat 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 confined primarily to lest aniicals of the 12 and 24 month sacrifices and they were dose-related in incidence and severity. % r* r:: In conclusion, Aroclcr 1254 does not appear to be carcinogenic in rats : fed.for .two years at levels up to and including 100 ppm. . j-.-t.ut.ii . .. ::_l 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: aew 58l67 ooo: Ay STLCOPCB4028957 IBT No. 622-07298 Monsanto 3 I have examined additional sections of liver from rats of EBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated my findings for Aroclor 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 cholangiohepatoma. The hypertrophic cells if. 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 058168 00Q STLCOPCB4028958 jJjd\Lzl B 1 O - T E S T $*c. 4 With exception of the vacuolar changes in the cytoplasm of hepatocytes, the '_ * more severe hepatocellular alterations were confined to animals of the 24- Month Sacrifice. (do/id Ward R. Richter, D.V.M. , M. S. Dipl., Am. College Vet. Path. SCH 058L69 01Q;'.77 STLCOPCB4028959 5 ' Lesion Primary Liver Lesions - Aroclor 1254 3 Month Sacrifice - Rats Cio'nn TI TE . TIE 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 2/10 _ 1/10 ____ 0/10 0/10 0/10 0/10 0/10 0/10 0/10 ' 0/10 0/10 0/10 0/10 10 0*3 G00:\~3 STLCOPCB4028960 6 Primary Liver Lesions - Aroclor 1254 6 Month Sacrifice Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular 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 Groun 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 STLCOPCB4028961 7 Primary Liver Lesions - Aroclor 1254 12 Month Sacrifice - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cholangio-Hepatoma Hepatocellular Necrosis n 1/10 0/10 o/io 0/10 1/10 0/10 0/10 ' th 4/10 4/10 0/10 o/io 1/10 0/10 0/10 Grout) 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 SCH .58U2 goq:`.~o STLCOPCB4028962 3 Primary Liver Lesions - Aroclor 1254 Terminal Sacrifice - Rats ' Lesion ' Vac-aolar 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 Gronp Tin 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 53113 " 000-' 31 STLCOPCB4028963 I Sacrifice interval * Month Tabulation of Individual Liver Lesions in Rats Dose Level Anima No. and Sex <n c o2 j >u0 2 o -3 0 c -roCct. CQ jQs. 's -O>. SS2ucu) (9 Uo -a~c. o. i =: -= 4U o kt u 3-1 lppm 3S6M 357" 358" 359" 360" 396F 397" 398" 399" 400" ++ + Liver Lesions zr --CL w OU U> r3 e4 CL CL > O 4 U o C=4 --4 "o CLL --> cX CL 5 -^C* *Q L 4-- 2 'o *O3 Z 4* <9 U4 -2 ~ O 9 2> 4 (J O C=4 4 wO 4 C3L X 4 o 4a xo 2tco 4 j2= u B-n lOppm 436M 437" 433" 439" 440" 476F 477" 478" 479" 430" B-m 516M lQOppm 517" 518" 519" 520" 555F 557" 558" 559" 560" 4 4 O O -- fw o l o C* t>. .2*j a 1 3c Cw co 52 *i* c <9 0 wj 2 SCH 058174 I! 010::.32 STLCOPCB4028964 STLCOPCB4028965 AHULLOft - 1 4 54 Tabulation oC Individual Liver Lesions in Rats Sacrifice I atervai l l i 2 Month Dose Level Amma No. and s *Sex 1Q v o2 n c .2 fr - 3 *4 31 2 0 1 jOa= 2 E J! Sc "2 6>. =- oa o >. u <U3 0 Cm <4J 0 Cm B-I lppm 41M 42" 43" 44" 45" 46F 47" 48" 49" 50" Liver Lesions oua 3u> M2 O idC. Q. >- i> iu0d Cm n 2; U >. S.8 --a Mo -- I- o Z <J 4 f 3 -- CL f 4U 0 m 4e o iaad. 4 E 04ao* 1 o "Sco "o u B-H lOppm 51M 52" 53" 54" 55" 56F 57" 58" 59" 60" ++ + + + + + B-m lOOppm 61M 62" 63" 64" 65" 66F 67" 68" 69" 70" Squamous C e ll C arcinom a, m e ta sta tic 1 SCM 058176 I 000:13 STLCOPCB4028966 STLCOPCB4028967 o^orcr, STLCOPCB4028968 I acrifice ni :rva,1 a Auuidcion Dose Level Anima r [ No. c and Sex _. - w 2,, o oa>*3. fwo> fl <a0 PC"fJ jrv. A o0 ?U 2u *a 1) C ---- cn rs a o- -^r au c^ -1 2a. o 0 <un 0 iu0s Ob & Liver Lesions 0 ^n --c. nu oU y>. T*4* 0 a a. n a. 1a3 -- e. t. o^ ' 3* 5 5* <0 i. r -- S 0 0 2' d w u^ 3 *r --o oQ, <v0 0 b 2 5* <oad a* * o <a0. JOi o *cto 2 u Month B-m lOOppm 484M 44 491" 497" 499" 507" 509" 510" + 512" 483" 44 489" + 522F 44 524" 44 527" 444 541" 547" 44 Extl" 44 Ext2" Ext3" 44 Ext4" Extn" 4 536" ++ 537" 539" 44 441" Exl" Ex2" 4+ Ex3" - . +P ++ P +P P. p + ++ + + + ++++ +++ 444 44 P P P P P P P P 4P 4' p 4 p 4 p 4 Grading System 4 44 444 4444 P = minimal in severity - mild in severity = moderate in severity a marked in severity = Present, no grade a Absent SCH 058179 Squam ous C e ll C a rc in o m a , m o ta ata t I -P G'TOr.C'? STLCOPCB4028969 STLCOPCB4028970 Jn&UtAud BIO -TEST Jldymtiyiies. 3nc. 1810 FRONTAG8 ROAO ,, NORTHBROOK. UllNOIS 60062 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. 64J - 06672 000333 STLCOPCB4028971 BIO-TEST . REPORT TO ' MONSANTO COMPANY 4 TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 -: _ IN ALBINO RATS HISTOPATHOLOGICA L EVALUATION OF ADDITIONAL LIVER SECTIONS : march 24,. 1975 IBT NO. 641 - 06672 . L Introduction ' . At the request of Dr. Levinska 3 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 ie E stained sections and evaluated by light microscopy. The following report presents the results of this study. SC* 058181 000:\33 STLCOPCB4028972 *ruLuUlal B l O - T E 5 T Jrt. 2 II. Summary In moat 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 datedt 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 m nature and they are morphologic manifestations of an adaptive response of the Ever 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: Manager, Toxicology SCM 058182 STLCOPCB4028973 BIO-TEST Jrux 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. my findings for Aroclor 1260 on the following pages: I have tabulated 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. SCH 058183 o-w ;x STLCOPCB4028974 jrulmi'JaL B I O * T E S T Iztxyiabyiiu. Jns. 4 With exception o the vacuolar changes in the cytoplasm of hepatocytes, the more severe hepatocellular alterations were confined to animals of the 12 % and 24-Month Sacrifice periods. hJcutJR.BuJM, Ward R. Richter. D. V. IvL , M. 3. Diplocnate. American College of Veterinary Pathologists SCH OSS'81' 000 STLCOPCB4028975 5 . Primary Liver Lesions - Aroclor 1260 3 Month Sacrifice - Rats Lesion . Grouo TX TII *THI ' 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 Duetula r 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 0/10 000:.43 STLCOPCB4028976 6 Primary Liver Lesions - Aroclor 1260 ------- 6 Month Sacrifice - Rats --- Lesion - Grouo TI TII Tm 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 Duetular 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 . 0710 0/10 1/10 SCM 058186 OOQlVU STLCOPCB4028977 7 Primary Liver Lesions - Aroclor 1260 12 Month Sacrifice - Rats Lesion * Grouo TI . Til Tni Vacuolar Change 2/9 5/10 3/9 Focal Hypertrophy 0/9 0/10 4/9 Nodular Hyperplasia 0/9 0/10 1/9 Hepatoma 0/9 0/10 0/9 Duetula r Hyperplasia 1/9 0/10 2/9 Cholangio-Hepatoma 0/9 0/10 0/9 Hepatocellular Necro sis 0/9 0/10 0/9 Control 1/10 0/10 0/10 0/10 1/10 0/10 C/1C l SCH 058181 000 i STLCOPCB4028978 8 Primary Liver Lesions - Aroclor 1260 Terminal Sacrifice - Rats ' Lesion * Grouo TX th Tin Vacuolar Change ` 5/25 . 6/23 10/27 Focal Hypertrophy 3/25 10/23 11/27 Nodular Hyperplasia 0/25 9/23 7/27 Hepatoma 0/25 0/23 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 V38 SC* OQQ'AG STLCOPCB4028979 1 ooo:v;7 ,, STLCOPCB4028980 AROCLOR 1260 Tabu tnlion of Inflivichi.il Liver Lesions in Finis STLCOPCB4028981 STLCOPCB4028982 Sacrifice Interval i Final . i ARCCLOR 1260 _ Tabulation of Individual Liver Lesions in Rats 12 Dose Level Anima { -- No. and Sex 0 c A u U "3o g > l 1 .a90* 0 u 9 2 *u 0 u "C o bai = ^c <0u & V0-4 ai uu t2 oa>S faoi js 0 Cu Liver Lesions >. a. _ uo u >* oa o >* j. 2* -33 0w O 2 is w It u3 --19 q 2--0 C9_ 23 > d0u ' fa 4 , c 0 ri Qm 2 ---- --------- o V i 0 ecj ft 0 u AI 85M 4 86" ++ 4 87" ++ 101" 106" 112" 114" 105" 107" 109" 4 122F 125" 127" 129" 130" 138" +++ 141" 147" 151" 155" + 144" 131" + 132" 135" 139" 4 4 4 _! 4 * 4 4 4 4 I 4. ah 167M 4 168" + 4 179" 44 182" P 184" ++ 187" ( Extra" 4 X* 205F P+ 208" fi. 210" -- SCH 058192 pOjO /- --O STLCOPCB4028983 AROCLOR 1260 Tabulation of Individual Liver Lesions in Rats STLCOPCB4028984 1260 Tahulntion of Individual Liver Lesions in Rats Grading System + = minimal in severity ++ = mild in severity . +++ = moderate in severity ++++ = marked in severity P = Present, no grade SC* 05B19<* ono:*.52 STLCOPCB4028985 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 His tops thological Re-evaluation of Tissues from Female Sherman Rats Fed Arodor 1260. As mentioned in this report, there vere about 20 animals vith lesions, vhich 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 I was not able to contact you by telephone. I was very much tied up with several visitors last week. . If you have any questions concerning the report, please contact me. Sincerely, P, Pour, M.D, Professor raj SCM 058195 UntMitHy a4 nmiu mmkii Cmr, * IM Oon AW, OciuM, U105 000^3 STLCOPCB4028986 i i \ E X H 1 B I T #, i STLCOPCB4028987 9n$jUjLt/ual BIO -TEST J?czJyyicyiie6. Snc. 1810 FRONTAGC ROAO NORTHBROOK. ILLINOIS 60062 . REPORT TO . ' MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1242 IN ALBINO RA* T% S ` HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS . MARCH 24, 1975 " IBT NO. 641 - 96672 2 DEPOSITION f EXHIBIT 1i 's-yi ti SCM 0581^9 000 STLCOPCB4028988 jrtdiuiual B l O * T E S T 3&yialyud4, Jrus. ' REPORT TO * MONSANTO COMPANY . TWO - YEAR CHRONIC ORALTOXICITY STUDY WITH AROCLOR 1242 - ' - -- IN ALBINO RATS............ ... HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS 7"iv ` ; L";'MARCH _24, T975-'-_ ' IBT NO. 641 - 06672 ' . .- L Introduction ** ' At the request of Dr. Leviaskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity study of Aroclor 1242 in rats (I3T No. 622-07298) were processed into H Se E stained sections and evaluated by light microscopy. The following report presents the results of this study. SCH 058150 000 jj STLCOPCB4028989 o I u l C J | ^a6o*a>tud, 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 axe 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 " ^ animals of the final sacrifice and they were dose-related in incidence and severity. In conclusion, Aroclor 1242 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, . Report Prepared and Reviewed by: D. E. Gordon, D.V.M., Ph.D. Section Head, Pathology Report Approved, by: fd ja*uager, i oxicoxogy SCM 058151 STLCOPCB4028990 Jtuiu.ii\ial tJ I U I fc 5 T jtalxyvzlosUcA, Jric. IBT No. 622-07298 Monsanto . 3 . - I ha/e 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 1242 on the following pages: * ' 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. SCH 58152 000 STLCOPCB4028991 JtvluAi'daL B 1 O T E S T JzA&vzLyUAi, 4 With exception of the vacujlar changes in the cytoplasm of hepatocytes. the more severe hepatocellular alterations were confined to animal* of the \ 24-Month Sacrifice. * (Jcuid&. R&ddh Ward R. Richter, D.V. M. ,M. S. DipL, Am. College Vet, Path. SCM 058153 o.o:`.33 STLCOPCB4028992 Primary Liver Lesions - Aroclor 1242 3 Month Sacrifice - Rats Lesion Group Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma TI 0/8 0/8 0/8 0/8 TU 2/10 0/10 0/10 0/10 Tm 1/9 0/9 0/9 6/9 Ductular Hyperplasia Cholangio- Hepatoma Hepatocellular Necrosis 0/8 0/8 1/8 0/10_ 1/9 0/10 0/9 0/10 0/9 5 Control 2/10 0/10 0/10 0/10 . 0/10 . 0/10 0/10 SCH 058154 ri t. goo:'CO STLCOPCB4028993 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 . tn 0/8 0/8 0/8 0/8 0/8 0/8 0/8- Croup - Tin 0/9 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 - ooor.GQ STLCOPCB4028994 7 Primary Liver Lesions - Aroclor 1242 I' 12 Month Sacrifice - Rats Lesion Grouo Vacuolar Change TI 2/10 TH ' 4/10 tm 0/9 Control 1/10 Focal Hypertrophy 0/10 0/10 1/9 0/10 Nodular Hyperplasia 0/10 0/10 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 SCH 058156 00Q1.61 STLCOPCB4028995 Primary Liver Lesions - Aroclor 1242 Terminal Sacritice - 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 Grouts TH . TIH 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 3 Control 1/23 0/23 1/23" 0/23 5/23 0/23 1/23 SCM 058157 0O0:'.C2 ^ STLCOPCB4028996 STLCOPCB4028997 ono:'.C4 ^ STLCOPCB4028998 AHU^LUU ------- Tabulation of Individual Liver Lesions in Rats Sacrifice terval Dose Animal] Level; No. and Sex 3 o>li *o-* S3 3o ^o --n. O ' o > oU 2n --e e o_: 2 C --<U4 CL .CO >. LL o C*4 2 Month C-I ! 71M Ippm j 72" 73" 74" 75" 76F 77" 78" 79" I 80" J.I I2 >t ~ 4U j--= O Cii Liver Lesi?nT J>O* OCL oca wU w> 5u c. o 21 IS a Us m 1Q. 9 3 >* n 2 --a ur o* 3 O o* 2-- QL. _J3 C9. u -- o I* eQ. O <a4 o M eC o O II c Lu0 r <n i-u O 32 32 w 9 L o ns > -- i, 'Jt fl --3 E s a' C-E ; 81M lOppm: 32" ; 83" 84" _ 85" ` 86F 87" 88" 89" 90" c-m : 91M 1 92" !-93" ! 94" i 95" ! 96F ! 97" '98" ;99" Control 1M 2" 3" 4" 5" 6F 7" 8" i 9" SCM 058160 - ono:cr: STLCOPCB4028999 aaouiacion ot individual Liver Lesions in Rats Sacrifice Interval M Month, Dose Level Anima No. and Sex 1a oo O2 3 t. 2 >2 2a O -<Zj <a9 2u *a. s71 jg 2c -C J5 o a. o <n c *3 O '3 " - (5 II c y__ C-I lppm 564M 566" 591" 567" .569" 572" 576" 580" 588" 589" 595" 60 IP 604" 605" 582" 583" 607" 608" 609" 613" 621" 627" 628" 631" 634" 635" 606" 610" 620" 626" 633" + + X a. au u ca>> *u a C*4 Liver Lesions --3 ua > a. -->* *a. u2 a " 1 o 2 xo aa = aU --9 OG. f (4 O U* <4 oP* aQO. a E0 4a 9 4eca 12 OU fc* o .2 S5 35 >. 2 SCM 058161- ono:*:cr, STLCOPCB4029000 AROCLOK - 1242 Tabulation oC Individual Liver Lesions in Rats 13 STLCOPCB4029001 O'lO/cn STLCOPCB4029002 Cytoplasm ic vacuolation of hepatocytes Focal necrosis of hopaiocytos Focal hypertrophy of hepatocylea Nodular hyperplasia of hepatocytes Focal bilo duct hypo rp la la C holanglo- hepatoma Reticulum C ell Sarcoma m etastatic Mammary tum or, ' m etastatic ARCCLOR - 1242 Tabulation of Individual Liver Lesions in Rats Sacrifice' nterval Dose Level Anima No. and Sex 1 n S 0c 1 c. t c~ 0o Li i Month. Control 63F 64" + 66" 70" 71" i: '' Ext" + Liver Lesions g 0 <oa9 * * P Grading System' + * Minim*3 in severity ++ = Mild in severity +++ = Moderate in severity ++++ s Marked in severity P = Present, no grade IS -- - X SCM 05816% oo: STLCOPCB4029003 9ru&jjLt/iAiL BIO -TEST laJ^yuiljyuzi,. Snc. 1810 FRONTACC ROAO NORTH 8ROOK, ILLINOIS 60062 REPORT TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1254 IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, IS75 - IBT NO. 641 - 06672 sc* 05Ql65 o'Tir/o STLCOPCB4029004 ILUX I B I T STLCOPCB4029005 f ( % * JhifcmlW Si O tist Jsioviami. Smi' 5 M* NONTMt *0*0 MOTMaOO.ILUMO<* *MI 221 UKtT TO MONSANTO COMPANT TWO - TEA* CHSONJC O&AL TCDCICITT STCBT WITH AAOCX.CC U34 IN A130(0 CATS KISTOPATHOUSCJCAI. ETAUJATION or AoomoNAi. uvxa sections MAXCH 24. 1373 IBT NO. Ml - 0M72 t DEPOSITION i EXHIBIT 2C.V|4ySErts44 I uo*j 000171 STLCOPCB4029006 JWfHlWf BIO-TIST 2aie*a29*ttA. %o. MM mtOMTAO* aoas MO*rNMOM.IUINOil Ml March U, trrs m Gaorfa J. Lavtaakaa, Ph.D. MmmiU Corapaay IOC N. Liodharfh Bird. 51. Louis, Mitioul &J144 Dear Dr. LeMaakaa: Ka: OT No. 641-0*672 Rlotopotfcolofical Eraloetlaa a( Additional Liver Sections from lui U a Two - Tear Chreaic Oral Trtcity 5tady at Aroelor 1254 W aro subrnltttng herewith our laboratory report dated March 14, 1575; prepared la coaaectlea with tba above etady. Vary truly yours. J. C. Calaadra President mi STLCOPCB4029007 '.V . ; MAW BIO-YSST ' * JL .V> '*&' -.A}*?- RETORT TO MOMSAMTO COMPACT two - rtAS aacrac asuu. toaoaxt rrnsT wrrx AJtocLO* um W ALSWO SLAT# RSTQPATHOLOGJCAl. ETALOATICK Or AOOrZXCMAL LIVES EECTIOW MASCH i\ 1973 1ST NO. Ml . OMTZ 223 At tha ol Dr. La-rtaafcaa tha Maaaaata Cmmpmry. aA&tioaal Mctlrn of Liwar {roes a two . ywar cknaic tnt tmrictty Maly M A roc lor la rata QBT Sa. ALZ-07Z99) wore procaaaa4 lata HAS aaiaa4 aactiaaa aa4 *vahiata4 by light microacopy. Tha foUaariag rapart praaaata tha rwaalta ol thia Italy. STLCOPCB4029008 IBT Ka. 422-0T2W I kaaa nirnimtnul nttlsu at Uasr tm rata at Mr Htaakar Ml*47294 ead tore ra<aeataa*a4 Mm far trUnn of aardaogoaosis. I tore whhaf mf fliMiftmiiiT far Arealor 1254 aa tba faOawiag pagaa. Tfcara ia aetfamoa at a otountoal afloat to fa Urn *Udi U moot pro* aa--cad at ttsa at fa yi-UaaA aad U-UtoH oacrifica. Tkia caaslsca at a fcopotocalhUar alteration kogiiwilttg aa local hrpatuogb/ progresses to aoifalor krvarplaaia. and ia a tom aaiaalo to bopatsma or ctolaagiakopatmaa. Tka hfpertiephla callo cawtaia larja oimmiwro at light otaiaiag cytoplasm vfclck La preh 'rip rick ia glycagaa ui aafaplaondc rortcaham. la him caaoo riag shaped itnmni, pretebty rapreaeatiag ohnU at proUfsrattse sadoplasmic retlnhaea. vara aaaa ia fa cytoplasm at thsoo sails. Tha kfptrpU<d< aodalas ontpM larger araao uf (Am compressed ..ttmxHt) colls at faa aarmai Uver parenchyma. They also caamlaed tfco oamo hypertrophic calls. Tkaaa laalaas which 1 claooiflsd as hepotamea or choUogla-hopeoomao vara larger aadniae which shews 4 oeidaaaa of coaflasaco or soms variatiea ia call sirs, akapa, staiaiag or a dactalar or adaaemateeo pottara at paotk. la fas aheaaca at metastasis, teeastoeaass. oarers fcassphllia. osteoses. or other ertdeaee at anaplasia; t chaos to call these fceaigo tamers rathor thaa ntaUgaeai (carcinomas). There was aa ertdsacs at motastasis or tasasieeaoes of these wttfc arrara ItafaMaaUaUr ittmAm i SacrlflM. U of tbs 14- Wax* ft. Uckasr. D.V.M..M.S. OtyU, Am. Callafa Tat. Patfc. " iju 00017G temmmeemr^e*:l. STLCOPCB4029011 5 227 Mawr ttmr Laafcna AmUt US4 1 Hurt iaartfiaa - Uh . ti TS l* - atm rm * Caacral YtwUr Oa*o focal lI)partToyhT Nartlar HyparpUata Bupatnnvi Dactalar Hyparplaaia ChaUagia Hepatoma HapatacaUolar Nacroaie 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 i/io qKU 1/10 0/10 t/10 0/10 0/10 0/10 1/10 0/10 0/10 0/10 0/10 o/to 0/10 000' 4 A STLCOPCB4029012 i Maurr ttror Uritu Amin UM . ' ^ " - iliaO Sacrifice ' tus ' Lloa Vacuolar Chine* Focal Hypertrophy Nodular Hypo rplaria Hepatoma DactalAT Hyperplasia Cty/Uagio-Hepacoroa Hapatecalhilar Nocrooia TJ 0/10 0/10 0/10 0/10 0/10 0/10 2/10 Tn 9/10 0/10 0/10 0/10 0/10 0/10 0/10 Ssa. TJB 1/10 * Control 1/10 4/10 ^ 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 l/io 1/10 STLCOPCB4029013 7 229 i PtlauT Um Uiteu Aroclar 1294 14 Hinirti SaariOca Kata i2i2* Yactaalu Chraga T ocal HTymoyhr Nodolar Hyyarplaala Hayatofpa Dactaiar Hj-parplaaia Cfcolaaylo-Bapatonia Hapatoca Ualar ttacroaia a 1/10 0/10 0/ld 0/10 1/10 0/10 0/10 Ta 4/10 4/10 o/ia 0/10 1/10 0/10 0/10 r*2SJHL Tin V Castral 1 l/io e/* 1/10 2/io *Ao 0/10 0/10 0/10 0/10 0/10 0/10 1/10 0/10 0/10 1/10 Vto 0/10 STLCOPCB4029014 s 230 Primary Um Laaiaoa Araclsr 1234 Tanoiaal Sacriflaa - &** Latloa Vtcwltr Qu( Tool Hypartroffey Nodular HrparpUaia Hepatoma Doctolar Hyparplaaia CioUa^io-Hspatoma Hapatocallolar Macroala 3 / 5 3 I 3 Oroap I Z l tHTX /ll Tt: V P t l/% 'fa 4 10/Z4 7*7. -ii/Z7 1 VtyA/ZS 1/11 / 5/Z4 yvtAf 13/27 Vj, V J/*J 0/11 0/11 4/zi 0/24 13/27 7 4/27 1/Z3 0/23 i/ll V 3/Zb VwA,- 4/27 vn\j 1/23 0/11 h.* 0/24 V 2/27 %.* i( 0/23 im V* V.U 1/Z4 11 2/27 MV 1/23 0a STLCOPCB4029015 KapaiMU /C lr rlol Sacrific* Iitaml * 3 Moot* A*OCLOft - UM TtWirtw ( MtoMwl Urmr Utlm la Km* D** HabD* ILt1 N. ul sHStx ii fi Jis.sJsUaai It H ii* ; J Ji 11 aa B-t l SOB 354M isr* j**" 3S7" 340" 3 HT srr3H" 344" <31 I B-n 434M 437" 438" 434" 440" 474T 477" 478" 477" B-D S14M lOO^ea sir1 SIB" 44 ! 17" SiO" Mir **7" Ml" nr* *40" **- t** .aJ.H in 5 * STLCOPCB4029016 AXOCLOC . U(4 Ttbatetta* mt tatftvMaal Uir UilMt ta Eat* to * frr:' , ooor.32 * W.II..li.A TV1irt|-- ' l STLCOPCB4029017 -v7^' - * ,: 2>v ' * v-' : /Vi-v-' \V AttOCLO* - UJ4 T*fcl*tia a/ ladtri4*al Lint Ultra la JL*fc 233 STLCOPCB4029018 STLCOPCB4029019 1 AR0CU3B US4 t3 STLCOPCB4029020 Io| m | i i i * / ( I r f h o a ii' .-i ' * AROCLO* - 14S4 Tt^iUrtwi C larfWt4**l Unr Li**g ta lUt* 236 u 3oc rifle* t^orrol Dooo Axima: Uni No. t tad Sox jj li iih u u 27 "3 r JL- Llror Loot-- m .2 Ua l e l | 1 3" o t i.f ij 24 Month B-m lOOpyn ab 4*414 r)++ 491" 497" 499" 507" $09" 510" 512" ** 483" ->+* 489" 522T ++ 524" 527" 541" 547" P*4+M Cxtl * * *-* Cxt2 bO 'H+* Crt4 SartSi 534' V++ 537' V !> 339' X* 441' Cxi* ZxZ *44 Cx3 W (-4 7)*4*4 b hp H4 w p H Pi-' p <n** t* * -J44 i* 87 H 0* Wn u.Pm ^4444 If.' t* <p P\^ vr P Hi* ^*444 W P H IP 4) P P *-> P W+* (f'W ie* *4 W P 4 P(- fiP w P t- iH Pi-* I) s- IH1 Crxdlnt Srttam - mintmol la oorortiy mild ia so-rartty 4 modanto la sonority 4444 mxrfca* la aarorl*y P ProMat, ao grado AIM* . STLCOPCB4029021 JU-'*W 810-TIST JiltiKfaMX. JU 2 237 .1 findings is tba lira r {ram this re-ewhastioe did cot differ algnifica: from that previously reported in our e igiaal report dated November 12, 1971. However, the-e vara aix benign liver.tumora datectad among at* of tba animale at tba highest treatment level (100 ppm) of tba 24-Mouth Sacrifice which arara aot prvvioudy reported. Tba otbar treatroent-reUted laaiona raportad ara regarded aa degenerative or hyperpUatie In nature acd they ara morphologic manifestations of an adaptive raaponaa of tba lirar ai)ci(ad wish biotransformation of the teat cratarlal. Tha Uttar laaiona arara confined primarily to teat animal* of the >4 and 24 month sacrlficaa and they arara doaa-raUtad in incidence and *va rity. In concluaion, Aroclor 1254 appeara to be (lightly tumorigenic at leer la at 100 ppm arLan fad continentsly la tha diet for taro yaara. Respectfully tuberuttad. INDUSTRIAL BIO-TEST LABORATORIES, INC. Report Prepared and R ariawad by: D. E. Cordon. D. V. M.. Ph. D. Section Head. Pathology ms7 STLCOPCB4029022 188 w IMuituai BIO - TIST ld**c2yv*. Jhc. ltfl r*OMTAO( *OAO . . sor>inao<.'uiNO( wi . REPORT TO MONSANTO COMPANT TWO - TEAK CHRONIC ORAL TOMCITT' STUDT WITH AXOCLOX 12*4 IN ALBINO RATS HISTOPATHOLOCICAL EVALUATION or AOOrTICNAL LIVER SECTIONS MARCH 24, irrs IBT NO, u: - 0M.72 /' STLCOPCB4029023 t Omitatml Bio.TltT . ifrjurrftMi. As. rtOMtAM ftOAO WO^TilHOOC, (UtMOti . . .t ? 189 Goory* J. Lavlaakaa, Ph-O- M H. Uadbartfe Bird. St. Leux, Mlmituni U1M D*r Dr. Lartaakaa: ZBT Ha. 441-Mi72 - Hlaaopacfcolotical Zraluattea al A^UtirKmal Lrnar Inrrlumii Fra* fate al * Two - Y*r Chrauc Oral Toxicity Study at Aroeloc 1254. W an eubsaittlA^ hamlth oar rrated laboratory mport dated Mmh 24. 1975; proparad la naiwrtei with tha ibex* ctady. Vary truly youra. J. C. Calaadr* _I STLCOPCB4029024 tmtntml tlO-TIST M BEPOBT TO MONSANTO COUPANT TWO - TEAS CXXGNIC ORAL TOXICFIT STOTT WITH A* OC LOR 1ZS4 IN ALBINO BATS ICSTOPATHOLOGICAL EYALCATICN or AsemauL uveb sections UABCH I*. UTS IBT NC. U1 . 06472 L Introdoctioa At taa n^ant ot Dr. Laaioafcoa ol tha Uoouan Coo-spaay, additional aactloaa of Lmt from & taro - yoar cbrooic oral toxicity arody of Aroclor 12S4 in rota (IBT No. 62.1 -77ZTO) vara prtsco a oad into H i I ataiaad Mcaoat and arataacad by li|bt microscopy. Tba fallowing rapor: pr .asta *a raaulta of tbia atudy. STLCOPCB4029025 ilO-TIST 191 n. la bmI SaaoacM, Aa apectnan a(t la tha Uar trsaa tUs n-whaha Ad aat i laputnad la our orlglaal report dsaad Wnaandini 12, 1171. aa-x baaiga lirr toamr* 1--acfad among as af tha wrtaala a Am high art Iraolnnat lrrel (100 ppa) at da 24 Maall Inn Mini vblck v M pi arWwnly royrtad. Om liver tumor (a hapawma) prarfaualy rtpnrfad to a 7-0 aafeaal (Ha. 445) Ml r*-claaatfiad aa aodolar hyprp4al Tha athar trinf mn ridma d laafana r.portad ar ragardad aa Jn|a>'iHn ar tijpat plastic la iiiiWm i and tbay ara morphologic naaliaatatiana at aa adapttaa raupmaa od tha Uvar aacrlbad la blotraaafarmatien al tha teat notarial. Tha latter laaiana wara rnnrtnad primarily to taat amlaw I a al tba 12 aad 24 month aacriScaa aad thay vara doaa ralamd mcidaaca aad arvsrtty. In rnnchiaton. Arcelor 12S4 appaars t> ba slightly namoriganic at laaaia al 10C ppm whaa lad CTntUwtiaty la Aa dint far two years. Respectfully nhsittida INDUSTRIAL BIO-TEST LABORATORIES. WC- Repcrt Prepared aad Reviewed 0. I. Cardan. O.V.M.. Ph.D. action Hand. Pathology Report Approved byi Id II. L. KapUapr^lk.oy Manager. Tadcalofy STLCOPCB4029026 a i BIO'TIST nr n*. tu-mn UwHntfl JU 192 I haw <rrmined additional sectlaas at Urn fresh rate at Stody Number 422-07294 usd haw re-ewtaoted thorn ter irltmca at carciaegeaesls. I haw tabulated my findings far A rex lar 1254 as the fallowing Tharo la evidence of a chemical affact aa eha Uwr which it moat pro- noanced at time of tha 12 -Uonth and 24-Month sacrifice. This coaetsts of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, aad la * low naimals to hepatoma or cholaagioSepatotna. Tha bypartrophic calls contain large amounts of light staining cytoplasm which ia probably rich ia glycogen and endoplasmic reticulum. la hoi eases ring shaped stractaras, probably representing whorls of proliferatim sodoplasimc reticulum. were sees ia the cytoplasm of thsse cells. Tha hyperplastic nodules occupied larger areas sod often compressed nrrouadlag cells of the aorma! V- "* ntrenchyma. They also contained the sarre hypertrophic cells. Thesa lesloos which ! classified ss hepatomas or eholasgto-hspatcmas were larger nodules which showed etrideace of ceafloeact or some wristioa in cetl else, shape, staining or a ductuler or adenomatous patters of growta. la the absence of metastasis, lnvaslwness. sere re basophilia, mitose s, ar other evidence of anaplasia; 1 chose to csll these beaign tamers rather than malignant i carcinomas). Tha re eras no evidence af metastasis or iawaiwaese of these minors la this study. . -' STLCOPCB4029027 With oscapdoa at tin ncnlu cfcaagaa la tb* cytapliam at hoyatocyfi, tha man Nnn lwpaMcaUalar aUantloaa war* comfl--rf tt Miami* at tb* Z4Mamth Sacrldea. (JmaJ A AUJ&f War**. tickttr, D.V.M..S4.S. DifL, Am. Catlap Vat. hit. STLCOPCB4029028 ..{&:&*v.'jtvAW:* * < t Vuwlu CM| rxtl Mfyrtroffcy . TMcff- 0/10 ~ o/w 1/10 z/10 0/10 Nodular Hrpsrplaaia 0/lfi 0/10 0/10 0/10 Btftnsa 0/10 0/10 0/10 0/10 Dact&Ur HyporpUaU 0/10 0/10 0/10 0/10 CieUn^io -Krpatona 0/10 0/10 0/10 0/10 V. HapAtooHular Nacrosla 1/10 1/10 0/10 0/10 STLCOPCB4029029 4 195 t Frisa&rf Lir*r Uiiou . ArocUr 1254 4 Mastii Swrlflct Ua taitoa Vicuolar Qiiaj* Toca.1 Hypertrophy Modular Hyp* rplieU HpAtoaa Ducraiar HyporplaiiA ChoUnjlo-Hepetotn* Hapto<aUular Necroal* TI 0/10 0/10 0/10 0/10 0/10 0/10 2/10 Til 0/10 0/10 0/10 0/10 0/10 0/10 0/10 Cna Tax 1/10 4/10 0/10 0/10 0/10 0/10 1/10 Coatrol mo 0/10 0/10 0. 10 C':a 0'10 l / 10 STLCOPCB4029030 Primary lioar Laaloaa Anxlor 12$4 12 Marti Sacrlfiea - Kata 196 Laalan Vae-aolar Qiaago Tocal Hypartropby Nodular Hyparplaala Hapatoma Doentlar Hyparplaaia Cho Uajio - Hapatoma Hapatocallular Nacroaia B 1/10 0/10 o/io 0/10 1/10 0/10 0/10 TO 4/10 4/10 0/10 0/10 1/10 0/10 0/10 2*2 Tm 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 7 STLCOPCB4029031 .117 Prtmarr Unr Lttioma - Aroclsr 1254 To retina1 Sacrifice Rat* L alon VtctaoUr Tjca1 Hypertrophy ;:o<iul*r HrperpUetA Hepatoma STtctular Hype rplijta ChoUa*io-Hepatoma Hepatocellular Nteteili Croczp TT Til TBX 8/31 10/26 33/27 3/11 5/26 13/27 0/11 3/26 13/27 0/31 0/26 4/27 o'31 3/26 4/27 0/31 . 0/26 2/37 3/31 1/26 zw c. W23 0.2J 1.23 3/23 5/23 0/23 1/23 STLCOPCB4029032 AROCLOR - US4 TabttUtloa oi iadivtdval Lhrot Ladotu ! Hats SacrUic* laterral Dm Lv*l 4nima No. f n1a.id Sox m U " 1 5} ii j"? &o 3 Smooth ! B-I ! lpptn t1 ; 1 356M 357" 35" 1359" 360" j 396r | 397" 1 39*" | I ! ; a-n !399" 40C" 43 614 'lOppm 437" f1 |i 43#" 43 9" 440" ! 476r i ] 477" 1 47#" | 1479" ! :480" I1 . B*EI 516M ;i00ppm.5l7" , 51#" ! 1519" ! 1520" 555r i '557" 55#" 559" 1 560" Lh*T Llotu * 11 j : If ft Ll 1 "m bs- U il :i 30 * Z 51 - f 1L ? 1 I . > 198 ai e2 n 11 I i! " .2 y I jI !2 STLCOPCB4029033 AltOCLOA 1254 Tabulation ot btdirvdmt Livar Lotion* in Rat* 5*crliic Doi A aim* lntrv1 ] Uni No. c Urtf L4ion 1 11 ind $4at 20 S* 0. S* , g 4 i: 5} ifi i i 1. is X1 3 * I! ii l * s1 11 ls Xi *a0 0 M>6__ Us. * 4U U i Z u il f1 4 8 l i 8 iu Month I> B-I i 921M 1 l ppm Q- .* i ;923" 924" 1 925" ,936F 937" ; 938" : 939" : 940" I a-c ' 95 0M 1 Opptn 951" 952" 953" 954" 966 F 967" 963" 96 9" 973" B 9J1M lOOppm: 932" i9*3" ;94" :9#j- .. I'96F |997 .998" 999" 1300" 10 199 * a: 3 ?! sS sl; *- STLCOPCB4029034 A HOCLOR - UJ4 TalmUti.0 at Ia4ivi4*l Liwr lxioaa la Kata fertile. latanrml Do Animt Uni No. ad j Sox 1 H \i a S If s H It Hi L ii fi.M5 "3 iif g i* *u5 W -3 'oo W Livar Lailoas 4 : 2 i. l tm * las i i ij ? I U Uonth B-t Ippm 1 41M 42" 43" 44" 45" 46F 4487"" 4590"" * lBO-paprn 51M 52" 53" 54" 55" * 56F 57" 58" 5 9" 50" * * ** " lOsO-mppm' 8o123"4 53" 64" I 65" ** 6667F" 68" 69" 70" * f 1 t H MJL i 3m fi .0 il >; UX8. U ta?~ b - jc 2 % STLCOPCB4029035 STLCOPCB4029036 AROCLOfl - 1214 Takalatioa o( ladlvtdual Llrar Laaiaaa la Rata Saerlllca Interval Doaa talma Llrar Laaiaaa nLara I No. aad Sax J S. o; si 21 21 a Si ! 4 is t* ii i-j LI 11 Si 5 , Jl -* 1 V 3 Iz 1 1 o ? 8 G 24 iXaath B-S 415U lOppm 417" I 1411" I 419" i j 423" : !*" . |427" | j491" I 402" j '<03'' ; i 405" j 1410" - I i " - ! j 450r I Mil'' ! 1452" 1 !454" ! !459" ! !443" I :449" j ; 470" ; 471" . ' 475" , 1443" , !445" P P 13 m. 0 i? 3! C* I 2 >. 1- il f4o 2"9 3S JT> ' ,dr 000202 STLCOPCB4029037 AKOCL<m - 134 Tabulation oi Individual Unt Lotion* la Rat* 14 203 Grading 3*t*m - imutiml la iTnry - * mild la r*rity --* mod*rmt* la a*v*rity *** markad la aowrlty P PrtMS, ao (rad* Abot .'iKjJ idSU allia` l-mIMiii 00G2Q3 L STLCOPCB4029038 V-/ ^s .O V.` 4* aro.THT " W* M*MTA MU ttOarwMOMM. llUNt MMI JW 2u--~-- * ' SSJKat TO MOMtANTO COMPANY TWO YZAI CKJtOttJC CSaL TOXICITY rrcoY with AJtCCLOS UI4 IX AU0 XAT3 KaTOPATXOLOOKAI. CVAL8ATI0N or AocmoNAj. um iecno< UAXCX 14. Iffl IS? MO. Ml 0**n A A / /!" STLCOPCB4029039 - ,i-.- Vi < !. ..;/*$ % ' ; * : '. N.. . oV.vrr Tf* '*' 7*1,, 'fX^Vv\-i:c-:':'- s'*-"' *. ' . . ' " ` <- .*v ' ' X - *\*.- .- .* j"* W- . 7 " . - ---------- ------- - : v ' ** r'~ r-Vlfi.H ...' V v;~.V} iV~* ^ i. .V . .t tliiiKftilii/^IO.TliT J-Ir 1. MB AdL/^l V >s;-. - * - > Oaarga J. Ml. M. LmMi .nui. IM. ttlM Dwlr. Sot BT Wa. Ml-64672 ttliiftiy irtwilngiml KYhatkn d fcdrfWncul Ltrar SocttoBS Jreea Kate of a Taw - 7r Cbrosic Aral Taaldty SafiuL^adaJSSl-------------------------------------------- Va ora --r*--"* Wuiartth oar* rvriaad laboratory rapert dated March 24,, 1VT5. prepar ad ta naaaartlna artdi dw ahava atady. Vary mly yaara. to . J. C. CaMadra .^gj/ JCC/td STLCOPCB4029040 W..W ftlO-TIST I y.^.y'-* .- :^- *.-:*% .v- -;.~;,..-*.: ;- ?~*. . .% ^V . - - V - 'i.S's-ar . '-*4- .- .-' ; ' **.. ..*; ,!*.-C*.\x ' . ` "*7. ' -r ^.n.' -a-nifif . 1 * iT - -`H JU MONSANTO COMPAXT TWO - TEAS CJOICNJC CSLAL TOXICITT 3TTOT WITH AEOCLCS US4 Ci AUDtO EATS H3TOPATHOLOCICAL EVAUTATICK or AflomoKAi. Ltrxa sections MASCH l*. im 1ST NO. 641 04472 - L Iatro<toctioa At tba rtfail oi Sr. Lavlaakas ai tfca Moaaaata Compear. aAditioeal octioa* oi lirar {root a tvs ysar circuit oral taaricirr tody #i Aroclar 12S4 - la rats IOT N- 6JI-072W) vara procaaoad into H k E **iaa4 xctioa* aa rral-oaead fay U4IU rclcroacopp. Tia iollov-La* report prM8tj a nrolu ci - chii ttaAr. STLCOPCB4029041 lO.TItf Ill, | Y< sis at t&a umlnla at *a Mflait (199 ppm) tf 6a H Month SaerlAca. Ka abaarad. Otm Unr tamr Ca hapaMmt ta a T*Q irtniwl Qb. 44S) vae n-dwtlM aa aoMolar hyparpiaala. Ha hypucplaaftt la imfaaa mi thaip ara morphologic arnnifiuKiimana ad aa adaptlva ad 9m &rar aacrtbad ta blotraaafanimtlita od da Mat malarial. 71m van aimflmini pa'laauUy to Mat aatamla ad the 12 ed M aaA fcay van <toaa ralatad la htcliianca aaA aa rarity. ta eaaaiaaMa. inda 12S4 appoaro m'ba akgfely't (-*- m" ,'-w* u ' jLi-f- "* < . . ad IM ppm vhaa ia4 raittirovfly la tha dhrt far Mra yaon. Baapactdollp atdbaUtad. MDUSTKUL BtO-TSST LAaOBATOMB. INC. Saport Praparvi tat lattaait bp S. X. OarMam. D. V.VL,Fh-D. Sactiam Baa A. Pathology a apart Afpmt Vyt M. I_ Kaptfa^ar, O. Wiaaagar. iMittltjy STLCOPCB4029042 br&a prateahly ia <ha Tin . at te t hyperplasia. Tin kf|WlifU( catb i ptitkMf rick ta **3 nd abaa Tiny aloe niHiiipm. irtnrtn^nm * Airtnim ar in ilium mmiihuiii pan-- at jwrt. b be abeanca ad of STLCOPCB4029043 o STLCOPCB4029044 STLCOPCB4029045 & 063 Primary Uv*r I^aiioaa Aroc lor US4 * Mostk Sacriflca Kata La* tea Vacuolar Qiaaja Tocal Hypertrophy Nodular Hypo rplaaia Hapatcma Ouctalar Hyparplaaia C ho 1o - H* patotu Hapatocallular Naeroata TX 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 Sees. ts: 1/10 4/10 0.10 0/10 0/10 0/10 i/io Control I/to 0/10 0/10 0/10 0/10 0/10 1/10 i i - `/'h >* mifip .......... STLCOPCB4029046 694 Prlauf Urv*z UriM - Amin 1254 12 Mart!k OiTlflr - Kata \r Vacuolar Cfetaga Focal Humitrophy Nodular Hyperplasia Hapaltoms Dietalar Hyperplasia Cholau*io-Hpatna Hepatocellular Nocrests S 1/10 0/10 0/10 0/10 1/10 0/10 0/10 th 4/10 4/10 0/10 0/10 1/10 0/10 0/10 2S9L Tin. Central 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 T STLCOPCB4029047 >j tony Ltm Laatom AmiM t294fi. ' / TaxiBlml Stcrlfica - ~ Modular HyparpUsl* Etyum Dvtslar Hrpn>lala QwUafia -Rapatoma Hapataca llalar Kacrarta 0/11 0/31 */31 0/11 3/31 3/1* o/i* 3/1* . 0/1* l/i* 13/1? *m 4/17 1/17 i/i? 1/13 0/13 3/13 0/13 1/13 STLCOPCB4029048 * S*criU jjf IotrrL 9 ) Usatfe AROCLOCt - U$4 T'VoiatJoo of MtvMtaoi Llnr Laaiooa to Rota m Do Loral B-I Ippa Ho. uU kx 3 iis*t 2asJ* 31 Mu f y_ 3S4M 357" 35*" 355" 340" 1W 357" 398" 355" <O0" Lirar Laataoa 3 1; If t ii it 1' 3 m' a ii hJr 1 * Jr ui B-n lOpjsn 434M 437" 438" 435" 440" 474T 477" 478" 475" 480" a-m 5I4M li I5 f*3S> : li STLCOPCB4029049 J 'mi n'oi. mmt ii . Z `Srftafcr.-J.., AaocLoe - usa j-- ToboUd-- of Mtriduil Livor Locioaa ta tot* iaerlfle* Litoral 4 XAoart Do* \nim Lor* l No. mad Sox 3 I* U f{ aJ ti u *l li U Is il t u_ s h> B-I Ippm 92IW 922" 923" 924" 925" 934T 937" 93*'* 939" 940" Unr Ltmiom fi 3 Is H -V: II11 if m i! bl. 1 1I rm UI B-H 950U lOppm 951" 952" 953" 954" 944T 947" 944" 949" 970" B-m I 94 LM I00ppmj942'' 943" 944" 945" 994T 994" 994" 1000" 067 ii I 10 t ' I STLCOPCB4029050 lk < |U tU ii/( lr r W > i` 'LL. . .5 * j ^ **"*v.V>.- - - Ay- - .;- ,<h Ur i. *idMUj*fii44 ' .-. 'i ri-;.*'* vT` *4ii')~ . .?..-'., -*** . ABOCLO* - U54 TmB%U*i*n ai iaAviAal Um Lt--* t ttti 018 U B-I *IM 42" 43" 44" 45" 4*r 47" 44" 44" 50" B-a lOppm i flU 52" 55" 54" *5" 547 57" 5" 54" 40" B-ca (OOppxn 4U4 42" 43" 44" 45" 447 47" 4B" 44" 70" | i i i i i i i i t I STLCOPCB4029051 j~Lm-- ^ AROClOa - US4 j u STLCOPCB4029052 " . ,+. . > '* AHOC LQK . US4 TibaUtioa ( ladtadaal Unr Lailotu la Xtti 07 is i STLCOPCB4029053 A HOC LOR - 134 Tabtalatloa / ladiridnal Uni Laaiona is Rata 71 14 Gradiat Syatam - minimal la uwrtty -- mild ia s-rarity ----- mod rat ia rarity *-- markad la ararity p Praaant, as pad* Abaaat STLCOPCB4029054 L* tioe Vacuolar Chiuag* Focal Hypertrophy Nodular Hyperplasia HffpfltomA Ductuiar Hyytrpkrta C Holinjio-Hepatoma fWpatocellulAr N<roi o m Primary Unr tnioat - Aradar 12)4 Tartniaal Sacrifice - Rata Tl i?3 3/31 5/2* 8/31 Til 12 Group TUI _____ 1 2 3 4/24 4/25 10/24 13/27* 3/2513/27 1/31 2/2* 3/31 1/24 1/25 5/24 6/27 3/25X3/27 S/2S 0/31 1/24 3/25 3/24 3/27 3/2513/27 9/2S /)l 0/25 0/24 1/27 2/25 4/27 3/31 4/28 4/31 3/24 4/25 3/24 5/27 2/25 4/27 4/2* 9/31 0/25 0/24 0/27 1/2S 2/27 2/31 2/2* 3/31 1/24 2/25 1/24 i/27 1/25 2/27 Control 12 1/23 0/24 0/23 0/24 1/23 0/24 0/23 0/24 5/23 5/24 0/23 0/24 1/23 0/24 1 - First nikttioo ot oriftMl slide a 2 - R*-evaluation erf orljiMl slide* 3 - Evaluation erf additional Haaua MCtioM Thera tra 5 animal* in Uch tMa lesion u not aaaa at tima irf IW aacoad mhrioa W wa prasaat on the first, ' Similar to above involving 4 aabaaU* . - STLCOPCB4029055 >aJS35i>:- ,\-1*.^!fe-'i-%j.s:-.'..- ;ss''.-jL. .-. . '/ <i'^*ii^itf(i%> ~fr ^Zfc;V''>VM^')r'.'~ i r'e^i&fc. riaii. f (O'TIST 073 Trimy Unr Utaa - A3ROCUML 1284 7T.~ __ TarariMt Smt -- A -* .: ** ^Itai %. .- ^ -*- oil._ Ov' $_ - . *-2 8-J - Cmrai Vic)v OlIMflO w 4/2$ V25 8/24 Fecal Hypainophy 2/23 1/2$ V2S 8/24 Nadvlar Byperylmrtii 0/28 V25 VIS 8/24 Ductalar Hyperplasia 4/28 4/2$ 2/2$ 4/21 2/28 8/2$ 1/2$ 9/24 Hapama 8/28 8/2$ 2/2$ 8/24 STLCOPCB4029056 UUiW 8IO.TIIT AI.hM .w - ' ' V ^*,, k,.pv -*..-^V. - -- - --.< - - - ^1,^ #74 STLCOPCB4029057 STLCOPCB4029058 otafe Vfc"~ - * *A i 1.1' l.tlThri' -- :{.- :r* * . n _ - t jjf^jfcTn'te'i. Uirnmtd BI0-T8ST /.tnanhim im. O 076 STLCOPCB4029059 STLCOPCB4029060 ',* %Autml 8IO-TIST /ffAwnAwat. JUe. ise *oNrot o*o 20 MO*THOOK. IILIMOIS MMI REPORT TO MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR W44 : .' IN ALBINO RATS KISTOPATHOLOCICAL EVALUATION Or ADDITIONAL LIVES SECTIONS MARCH 24. 1975 IBT NO. 441 - Obi?; "2 <C STLCOPCB4029061 Mtktfual #10 -TUT JabovtbvtH.. 3k. 205 1 MIC *OwTACt *01-0 Moimttaoot.auKOil ***a Goorgs J, Loriaahos, FfcuD. Vionsaoto Compaay 800 N. Lindbargh Bird. St Louis, Missouri 63144 Duar Dr. Lsuiaakas: R: BT No. 641 -06 672 Htotopatfao logical Enlunoa of Additional Liror Soctioas From Rota of a Tuo> Tour Chronic Oral Toxicity StorfT of Aroclor 8S4S. W are submitting hsruaritb our1 laboratory nrport dat*d Warefa 24, 1975. p^oparod la eooaactloa sritfc tbs aboro study. Vary truly yours. J. C. Calaodra Prosid* at STLCOPCB4029062 jUn<*>*( BI0-T31T JU 206 RITOtT TO MONSANTO COUPAXT TWO TEAS CHRONIC ORAL TOXICITT STUDY WITH AROCLO* H4<M.O IN ALBINO RATS KXSTOPATHOLOGXCAL EVALUATION or ADDITIONAL LTVER SECTIONS MARCH 24, 197! QT NO. 641 - 06672 L Introduction At the retjeart el Dr. Lerhaakaa el the Moeaaato Compeer, additional ilit eectioea of lira r (rem & two - yaa r chronic oral toxicity rody of A roc lo r Ml in rata (LBT No. 622-07298) were procaoaed l*to K 6 E ataiaed aaetioaa aad evaluatc-d by lijht tclcroacopy. The Iclloarinj report praeesta the reenlta of thti atudy. STLCOPCB4029063 207 he ttver treat this re-evalaatosa did bM differ eigalBcaatty frsm that previously r*p(m< to ear niflitl repert datod Na*t 12, H71. Hmm, there were eeveo haaiga liver tamers detected un( eovgp of toe pel mila at toe highest treeerneat level (104 ppm) sf toe 24-Meath Sacrifice which were set previously reported. The other tTPatmeat-related Leetoe* reported are regarded ai degenera tive or hyperplaetlc la setare aad they are marphologic maaileetatioaj of as adaptive reepeae* ol the Uarer aeaociatod with hiotruaiomatiea el toe tet material, la geaeral, the latter findings were ceafiaed primarily to tagt animals t ti si the^pd. )rtT~ cad 24-Mnnfh Sacrifice*, aad they were deee-releted to incidence aad severity. ' i; - la csseiiuloa. Aroclor hidd-eppeers to he slightly tumerigeaic at levels of 100 ppm whea fed conttaooealy to toe diet for two years. Kespectfully submitted. Sleport Prepared sei I 0. C. Gsrdea. D.T.U..Ph. 0. Secttoa Head. Patoato^y MM IIO-TIST JU.a*.* Jm. 1 1ST IW. 4M-C7rt* { Mwnte 208 I Wn mnliad Mltloaal Mcttea* af Utt tr*e rats af BT No. 422-07278 oaf fears re-ralta4 tfeota ter rrMnaa af aanlaag--mIi fesrs t4 mlnllmif Bkf Bite|i ter Araclar - Z& m tfco loUewtag papost I There la evidence f ft chemical affect *a tho Ueer aUck was ott 12. * evident ftt time af the rdrhteBth and terminal (24-fcfemh) sacrifices. This cociiffU f a hspatocellolar alteration haplpafeg as fecal hypertrophy which progresses to mnitnlnr hyperplasia. and is a few animals. to hepatoma* or cholaagiohepatomas. The hypertrophic calls erafata large amsont* af lightstaining cytoplasm which is probably rich ia glycogen and endoptoantic reti culum. la some caoas. ring-shaped structures. probably representing whorls /- of proLlferativs endoplasmic reticulum, wore oom ia the cytoplasm af these eUa. The hyperplastic nodules occupied larger areas aad often compressed surrounding cells of the aorm&l liver parenchyma. They also contained the same hypertrophic cells. . Those lesioas which I classified as hepatomas or cholaagiohepatomaj were larger nodules which showed evidence of confluence or seme variation us ceil sise. shape, staining or a doctular er adenomatous pertera of growth- la the absence of motasted*, lavasiveaess. severe basophilia, mitoses or other erldence of anaplasia. I chose to call these benign tumors rather than maLignan: carcinomas. There aas no evidence of metastasis sr Lev*stveae*s af these tumors ia this study. STLCOPCB4029065 W 8IO-T88T ' 209 --.--> ^u-frnlamt W liw> ixifcn wlhl|<t nri|ht H-*n Mim--^ nwiiiriii1iiiiiiimwib mu Htn fcrnrtn Tf'Htnrr i r" "`i t*t` 1111 iniflir^***** *--** k* Wltfc miinulln * ths VMtaoUr ckaaf** U tks cyMfUaa *f **<yt. th owra **vr fc*pfteUalar altaratiau *ra * 1_ M atiml *1 tha H: ml 24-Mntk fcerifln ytrMt. UUaJQ . &*JMi Ward R. UcKtar. D. v. M.. U. S. Diplomato, Amorlcaa CdlUfo of Votort&ary Pai^oLojiits "V-tL :vi. 0002;if STLCOPCB4029066 s . 21 0 * lv Primurr u>r Laiu - Arvclar IMS 3 Uaad l*trlflc< lU Own VftewUr rocal gyportrophy No^ai&r RyprpUU Hp*tocaA Ductal* r HyprpU^U OvcU-afto H^|Hitocrm H4p*.tocllL&r Ncroois TX 0/11 0/11 0/11 0/11 0/11 0/11 0/11 Til 4/10 0/10 0/10 0/10 1/10 0/10 0/10 Tttl Ceatrot J/10 7c 2/10 2/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 STLCOPCB4029067 >6 2)1 s> * Primary Unr LmUu Aroelar U Mantk SuriSct Slats ias.op Vacuolar Chaafa focal Hypartrophy Modular Byparplaala Ha patom* Oww TI Til Tin Control Mb i/5 5/9 c/*>. t/10 0/6 0/5 3/9 V, 0/10 0/6 0/5 . C/9 0/10 0/6 0/5 0/9 0/10 Duetular Hyparplatla 0/6 0/5 0/9 . 0/10 Cbolaaflo -Ha pa torn* 0/6 0/5 0/9 0/10 Hapatocallalar Macresis 0/6 1/5 0/9 1/10 STLCOPCB4029068 *) 212 l^rixaary Utt Loaie** Arador U4i 14 MHalt S*rtflc - KsU La sloe Vacvalar Cteag* Focal Hypartroyfey fey? th Tin Coatrol * 1/9 5/10 1/9 1/10 0/9 0/10 4/9 0/10 Kodalar Byparplaata Bapatotm Doctalar Byparplaaia 0/9 0/10 1/9 V 0/10 0/9 0/10 0/9 0/10 1/9 0/10 1/9 1/10 Cbolaa(lo>BapacscDa 0/9 0/10 0/9 0/10 ) Bapatocallalar Nocroaia 0/9 0/10 0/9 0/10 STLCOPCB4029069 s 213 fro Priam? Li'rar La stoma Aroclar 1242. Larisa Tonaiaal fecrlfica - 1UU Qro^ Vac-no Ur Ckaaf* TocaI Hyygrtioyilif Nodular Hj-parpUaia HftpfctMB* Ometalar HjrparpUaU ChoUaf io-Hi^atoma KapatocalhaUr Nacrosio *V 9f */ Tn A(. 6/23 nn %*. Ceitnl \*u10/27 1/23 .-W / s/2S v'4' 10/23 *U^ iV27 Y,.*;., 0/23 A- 0/23 0/23 "V 5/23 tu' 0/23 7/27 1/.3 V 3/27 7m 'n 0/23 ,/,v tj; v 6/23 ',r>-c 5/23 Va 14/27 /.- ! 3/23 d/>f 0/25 "a,' 0/23 2/27 /.- f.. 0/23 '/'/ 4/25 %.t 2/23 ''ll '</ 4/27 V..- 7., 1/23 f * f. <f--. * >* >- > ,v ^ * v> ^ 0n0Z3fl || STLCOPCB4029070 AJtOCLC 124*^ TaUqUUm of fndivMn*! Liver Letlofu la Kali - 2I 4 'Ucrtflce Uttrval * Dm L*l ^nima No. oixl *u u 1 a > m Vs 1 o X lit 3 Mooch A1 . T . 7>. c 11 AW 117" 115" 117" 120" 147T 154" 157" 15" 157" 140" Un r Latu * f: si fr t H 5* 1XA *k* H li li 5* 3 iiS U ftV0fe Z 5l h i rj j X0 u 0 AC 175W 4 174" 4 171" 200" Extra" 2341* 237" 235" 237" 240" Ad -'t t/r 274W 277" -4+ 275" 27?" 250" 3isr nr' 315" 31?" 320" <-* 4 i-' 44 55*" f**--* -l rwr r$ %" * ^*sr**-i* *'- ^ m?fsTh* .V ** OOO.^.r: STLCOPCB4029071 AJLOCU38. UU Tabulation *1 InriWMosI Unr l-a*ion* hi Rat* , 221155 10 tor1 *;* ' \ ^vS*v; \'"fc 'iil. T^. 000236 STLCOPCB4029072  Aiocxx* ' Tabulation of individual Liver Letiou la Rats 216 * STLCOPCB4029073 ` ' * * i. 1 *> ** V`m. 0007-38 STLCOPCB4029074 UOOOK Ul* 13 jaOCLCBL 4& Tibotatlfwi ( Individo*! Unr Lwim In Rtu 2'9 Gn&as Sratam *+ *** P > mtatmal in amrlty mil* ia avmrtty * taaianta ia *-*riry nark4 ia severity Phhk, to |rvli yftSf STLCOPCB4029076 JftSusiucU 810 -TIST 11C MONtAOl *0*0 NO'T"l*OOr.iUWOH *0041 She. REPORT TO MCN&uNTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH ARCCLOR 1260 IN ALBINO RATS HISTOPATHOLOGIC A L. EVALUATION or additional liver sections MARCH 14. 1975 I3T NO. 641 - 06671 172 STLCOPCB4029077 %&afaal ilO -TIST Jao*\t<n*i. Dnc. WO **CWTAO* OAO MOTMlOOC. tlVtNOtft +0001 173 Cwrgt J. Loriaskaa, Ph.D. Mowntp f nsiqurty #00 N. Llnrfbryh &lvd. Si. Uwns. Miaaouri 631fc4 . Dmt Dr. Lretnakao: Re: IBT Ho. Ml-0fe472 * KiasopatfaologicaJ Er&luatkie d Additional Litot Sections Frets Rots oi a Tsn> * Year Chrome Oral Toxicity Study erf Arpclor 1240. W are subsait&ng baroartlh our rarlsod laboratory report dated March 24. 1775: prepared ts mn--rrlnn with the abora study. Vary truly years. J. C. Calandrs Prasndont STLCOPCB4029078 ftIOTI ST C Am. in report to MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITY STUDY WITH A ROC LOR 12*4 IN ALBINO RATS HIST OPATH LOGICAL EVALUATION or additional liver sections MARCH 24. 1975 IBT NO. *41 - 04672 I. Introduction At tha raqtaaat wl Dr. Lariaafcaa oI tfa Monsanto C0fr4.it'', additional aaetiona of livar frost a tiro yaar chronic oral toxicity study of Aroctor 12*4 in rata 'ITT No. *22-07298) srara procasaad into K * E stainad aactions and avaluatad by lifht microscopy. Tha following raport praaanta tna rasulti of this study. rrr< STLCOPCB4029079 2. Q. SuMMOTT b aoat lutuctt, t'aa spectime at trpatmant-rslsiaid hf stopsthiubgknl fiedbgs ta tba Unr from this ra-erahiatioji did Ml differ significantly from that prerioosly reported la our original report dated Noromber 12, 1971. Hoveeor, than vara seean benign lira r tamors dataclad among aarca of the at the thghest treatment la-ral (100 ppm) at the 24-Uanth Sacrifice which warn sot pranoaaly reported. Tba othar treatment-related lesions reported are regarded aa degetasra- tiaa or hyperplastic is natore aad they ara morphologic manifestations of aa ndaptfre response of tba lieer aaaociatad with biotmnsformation of tba taat material. In general, tba Uttar find Inga atara coofload primarily to taat an) mala of tba 6*. 12* and 24-hionth Sacrifices. and tboy vara doaa*ralatad La incfdanc* and serenity. la conclusion. Aroclor 1760 appaara to bn slightly tumorignoic at larels of 100 pprs arbao fed cocdouously la tba diet for taro years. Respectf-olly s-shmitted. INDUSTRIAL BIO-TEST LABORATORIES. INC. Report Prepared aad Reviswod by: D. t. Gordon. D. 7.U..Pi. D. Section Hand. Pathology -'-ns'*--*w 'BQtJJPff T -*T- - -jgfc- ipr.sr- ***r SIO-TIST & IBT No. 4U*07Z?t MaaMBte 176 l her* aMUioaol aoctlooj ( lir%t tr*ca ntt < OT Ho. 4iZ-OTMS aad bora ro-srmlaMsd thorn tor ridsca at ca-rriiwigimnuH.*. I v. bare tabtUu^ my fto4ta^s for Arodor m tb feUovtag pages: There is evidence of a chemical effect o the liver which v&i most evident at time of the 12- Month and terminal (24-Moatfa) sacrifices. This consists of & hepatocellular alteration beginning ss focal hypertrophy which progresses to soduLsr hyperplasia* and to a few aidmail, to benetomas or cholsngiohepatomaa* The hypertrophic cells contain lar p amounts of U%h: - staining cytoplasm which is probably rich la glycogen and endoplasmic re:.' cuium. 2n some cases, ring-shaped structures, probably representing who.-Is of proliferative endoplasmic reticulum. eer seen in the cytoplasm c: these cells. The hyperplastic nodules occupied larger areas and oftsa compressed surrounding cells of the normal lWr parenchyma. They aLee contained the same hypertrophic cells. Those lesions which I classified as hepatomas or chelangiohepsrorr.sa were Larger nodules which showed evidence of confluence or some variar.or. ;r cell sise. shape, staining er a dueruler or adenomatous pattern of growth. In the absence of metastasis. LxrrasWeness. severe basophilia, mitoses or otr.tr evidenee of anaplasia. I chose to call these benign tumors rather than malignant carcinomas. There was no evidence of metastasis o. leva*imoots of (bi rumors in this study. STLCOPCB4029081 JUSIO-T8ST Afa*k<m. &w 4 ' 177 With sgcapOon af tbs racwolar cba| is tfea r ymiplmm at hnpM l jIiiiii , r.-fr" . : <b) Bar* mai kafOaoBsltr iharukai war* Tiftnfit to sataals at tka U- tad 24-Meatb SacrtUca paitoda. ' ud Hichtor. D. V. U.. M. S. Diplomat*. Amariean CoU*g* of Vetansary Pathologist* STLCOPCB4029082 1 Mouth Sacrifle - Siti L**loo VicmU: Chaa{ focal Hyp rtropfey Nodular Hyparplaaia HopOtOtn* Ooctular Hrparplaaia QioUajio -Hepatoma Kapatoeellular Nocroaia TI 0/U 0/11 0/11 0/11 0/11 0/11 0/11 Til */io 0/10 0/10 0/10 1/10 0/10 0/10 TUI 3/10 2/10 0/10 0/10 0/10 0/10 0/10 Coctroi 2/10 0/10 0 !0 O'lO 0/1C 0/10 0/10 STLCOPCB4029083 Primary Liver Lestoae Araclor 12M t Uodh beriflci - tail La elan Gre-ni Vacuolar Changa TI Til ns 1/6 2/9 5/5 Tocal Hypertrophy 0/6 0/5 3/9 Nodular Hyperplaala 0/6 0/5 0/9 Hepatoma 0/6 0/5 0/9 Duerular Hyperplaala 0/6 0/5 0/9 Chnlanfto-Hepatoma 0/6 0/5 0/9 Hepatocellular Necroale 0/6 1/5 0/9 Control 1/10 O'lO 0/10 0/10 0/10 0/10 1/10 STLCOPCB4029084 7 Primary lira r Laiumi - Aroc lor 12M U Moath SacriAt* Kats Laaion SS T1 Tn rm Vacuolar Claanja focal HtPrtrophy 2/9 5/10 3/9 0/9 0/10 4/9 Nodular HyptrplatU H*p* COCOA 0/9 0/10 1/9 0/Q 0/10 0'1 Duct&lar Hyparplaala 1/9 0/10 i/9 ChoU&fio* Hepatoma 0/9 0/10 O/o HepatocolluUr Ncrosl 0/9 0/10 0/9 no Control 1 -'10 o/io 0/10 0 '10 1/1C o;*o O'lO STLCOPCB4029085 .... '.-Gtfi-v 8 s .. m Primary Uwr Laaioaa - A.rocior^t3M ife' lr ' -- *- ' Tan&lml Sacrifle# lata v- v? - * 4_ ' Vtneur Chug* T ocal H;yrtroyti) NcxJuiar KyparpU-sU Hapatstaa Dnctnlar HyparpUaia ChoUngla-Hapatuca HapatocaUular Nacroou t: sm 3/25 0/25 0/25 4/25 0/25 4/25 Tn 4/23 10/23 9/23 0/23 5/23 0/23 2/23 TM5 Tm 10/27 11-27. 7'27 5/27 14'27 2/27 4/27 Control 1/23 0'23 1.23 0/23 5/23 0 '23 1/23 STLCOPCB4029086 Vfr' ;> 8t-.fr; 4 /-Y'*- '!!;. ,r .. "'T A Tfi*i mSSL*; AJIOCIX* UM T^UnUiieflW MUMitfl L'wr Leal*** la lUlf 182 STLCOPCB4029087 1 V sA ^ ofTalwUllen uocua um Individual Liver Loaion* Rat* Sacrifice Interval Coh Level \nima No. and Sex Liver La*lop* Ifv S 0 1: II If t! Li J l 4* =1 6 Month AI S3ZM !*J4" 1*35" Isi&r 1st*" *49" I .'An *4M 65" *73" iSTST l79" 0 h. Zio . Affi 8 9 lit *93" *94" *95" 19067 I - 1907" i908" 909" 910" 183 10 :IJ r i U STLCOPCB4029088 J l Jaii AXOCUM UW Tabnlnlion W Individual Liver Le*lo* ia IUU 184 ** STLCOPCB4029089 :t Xkl 4 c STLCOPCB4029090 000255 STLCOPCB4029091 JL JAOCVOO. 12M TftbaUOon of WH>i*al Liver Utiou la Itau 187 14 System * ml alma 1 la aevority ** mOd la sorority ** moderate la severity +** marked la severity P Present, as (rads wag*-'** * " ` v * ` ' csfr'*"'vs <T . - "i . ' ... **, .f.*~' ' onor*5G STLCOPCB4029092 MtUUmJ, ftIO *T8ST jSxttaM*. She... ie ftoxrAoc ftoxc MOTHr*OOC.tlVINO<* tun 077 REPORT TC MONSANTO COMPANT TWO - TEAR CHRONIC ORAL TOXICITY STUDY WITH A ROC LOR 11*0 IN ALBINO RATS ICSTOPATHOLOGICAL EVALUATION Or ADDITIONAL LIVER SECTIONS MARCH 24, 1 975 1ST NO. 441 - 0ooT2 S , 41 4\ / .a STLCOPCB4029093 JU&at&d tlO-TtST Jo&Malaxeab. %a. 1IM HOMTiOi *0*0 MomuooLiuiMoa tom Jly 1. It75 078 Goary* J. Lrrtaaku, Bi.D. Momtamw Company *00 M. liadberjh Bird. St. Udt, Missouri U1M - Deer Or. Levtaakaoi BLa: 1ST No. Ml-04*73 - KlatopatholoQtcal Kvaluatie* of Additional Liver Section* From Kata of a Two - Tear CJomic Oral Tortclty Study oi Arodor 13*0. We are eubndtttnj bareertth oar revload Laboratory report dated March 24, 1475. pnpond lit connection vltii A* above atady. Very truly year*, Xo . *So * -fiat -i y S. C. Galaudra JCC/d 1. STLCOPCB4029094 W-W SIO-TIST 97$ BEPOST TO MONSANTO C CMPANT TWO - TEA* CHRONIC ORAL TOXICITT STUDY WITH AROCLO* 1260 IN ALBINO RATS HE5TOPATHOLOCICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24. 1^75 IBT NO. 641 - 06472 At the raquaat of Dr. L*ink of tfao ilooconro Compter. additional ction of iivor from a taro r** chronic orl toxicity atudy of Aroclor 1240 in rtu I LET No. 622-07245) woro proeoaaod into H k E atainod loctiona test avaluatrd hr li*ht mjeroacopy. Th* .'olloarinf roport prnt :h rornl:* of thi i *tudv# STLCOPCB4029095 r. ' tfi Idt.fehf 8I0.TSST e B. --rt -r-w 2 089 fiadtaga hi fee Mver ftaa fete wtwlufea Ad net dttbr id|sflflmiiit1j tea feat piartealy t sported h aar hiflail rapmrt dated Wiwumlinr 12. Wl. Thera in wwa hapeteaaa detected among arraa ad fea ateali at fea highest traatrwimt Wrel (1W pfaj at fea 14-Mate Sacrifice. Mo hepatocellular carcinomas wi abasrvsd. The ntHiimi1 trostisnt ralitad lesions reported ara regarded aa degenerative or hype rplastir la nature aad they ara Morphologic BMal/aatattena at aa adaptive rcapooea of fea tivar iiMoctatod wife biotraneiovmatloa at fea teat asterial. fat general, fea latter findings war* rraflaod primarily to test ol fea 6-, 12- aad 24-Month Sacrlficaa, aad feay man doae- retetod la Incidence and esrarity. Is conehasten. Arecter 1260 dons net appear to be carciaogtnic in rats ted ter two years at Wmls sp te and feel tiding 100 ppa. Respectfully subedited. INDUSTRIAL BIO-TXST LABOSATORISS. INC. Report Prepared aad Reviewed by: D. E. Gordoa. D. w. M.. Ph. D. Ssctioe Hoad. Pathe logy Rapert Approved by: 000PG0 l > l_ STLCOPCB4029096  :r: I?'.. .... - - - :..j* .-5* :a c'"T-..: .'. K -W'- ')' ` f.r/ .^.` -.* r1.- V--- w`..~ - -.?V <.%; -<'*; -V4t.?/..'-; -t.-P~!"-v3K-.AV^'-''.', '.! .. . ..-, .-" v*. ' ' ;|>1 vf: - *. ;_xv - .; i- ,, *. '>'-/ A:Xv~' .. : :- _ ..tiw e J3iCA*lLj^&2ix}i ^ 1^ * - - - M~ -*- *. vr~ 10-TltT o BT Ha. U2S721Q -V: .. - *" * ` *< lw w dmH*. ua-*rm kw tar iiwjiimiiffl f UM m ft* Man* .'11 ts art taw ef a chiwriml affect os fea Htw which mat aaidaut at ttm ad 0m UHtaA and hniul (lHhafe) uoiflcat. lht tinntai od a hspatocottalar altaaartos bsgtaatog as fecal hypartrophy which pragraasaa to nodalar byparplaata. and to a tow soiaMla. to hapatn-- or cbslasgicbapaaosu. Tha hyparfeophlc call* caatata larg* a--pm at light stsistag cy-upUam which U probably rich is glyeogsa sad endoplasmic rstUubms. la soma CUM. rtag hspad structures, probably icprowes dag wborla at preiiiaraiiT* mdnplosmic reticulum. ware seam ia tba cytoplasm at that* calls. Tbs hyperplastic amduikai accepted larger araas sad odtaa rsniprwin id surreusdisg calls at *a .... ....ill haar parenchyma. They slsa rianhid tha seam hypertrophic cUs. Thaaa laaiaaa classified H hep unman or chalsiigiahepsimiMO wars larger ooduloa which shawad aridaaca at ensdlamca or sobs aartsdsa Is call sise. shape, or a dactalar ar adenomatous pattorn at giwwth. Is tha ahaataca at mtaatuia, brresieessese. --ears basephill*. evidence od muplsaia. thaaa ora benign twn rather 0 carciaoma. Thora was no avidaaca at matsstasia or iavasivtins at thaaa rumors in this study. \gn& ypiQ^ip |fF> aWS 002Si STLCOPCB4029097 4 3< Mwirtl Bum Kin pMi. t to attaals cf Am 12- W%x4 L tlcter. D.V.M., U. i Pt^inxm. Amncu CoUf o TtwrtMry PatholOflK* a - - 'HfTi V- ~ .* /V7 `y.** * 000262 +m+ STLCOPCB4029098 { V' . *c: * - r- . ' Vv $ \ ; l' * / frliwHTy Um Latina Amlar I MatthoiBct - lUts . ;-A*r / . v - ` SOL '' VicboIat Oup rcAi ary*ttu>fcr MeMtr SyparplAiiA Hapatxrm& OoetaUr HypBrpIaaIa QtoUajio BpAtemA Kap&tocaUalAr N Croats Tt <9/11 9/11 0/11 0/11 0/11 0/11 0/11 : 4/10 0/10 o/to 0/10 1/10 0/10 0/10 rni 1/10 2/10 0/10 0/10 0/10 0/10 0/10 Ci--tr<il 2/10 0/10 . 0/10 0/10 0/10 0/10 0/10 I t STLCOPCB4029099 * A- , Pwtutrr Umi lHw Andtr Ota * 4 MaoCh SMTiflca - liu lU Vacuolar Ch&age Ow XI TU tux 1/* 2/5 5/4 foal Hrytftnfkr NoMu 0/4 0/4 0/4 0/5 J/4 0/5 0/4 0/5 0/4 Doctalsr HyparpUala C^almio-Hapatoma KopuracaUuiar Ncroots 0/4 0/4 0/4 0/5 0/ 0/5 0/4 1/5 0/4 CoatroL 2/10 0/10 0/10 0/10 . 0/10 0/10 1/10 STLCOPCB4029100 X... C&5 Primary IJrar Laalaau - Aroclor 124# 2 Meath SacnOca - Uu Laale* Gm^ TI T3 TCI Vaaoolar Cfcaaga 2/9 5/10 3/9 rocal Hypertrophy No<hiUr HyparpUaL. 0/9 0/10 4/9 o/* 0 '-.o l. Hepatoma 0/9 C/10 0'S DoctaUr HyparpiaaU J/9 0/10 2/9 Cbolaa^io-Hepatoma 0/9 0/10 0 Hepatocellular Macro ala 0/9 0/10 0 Ccetrol 1/10 0 10 C 10 o -;o 10 3 10 3 :o STLCOPCB4029101 I Primary L*aiaa Ancln UM Tarmiaal iaertflaa - Rats Laaioe '* Om . TZ TB TB2 Vacaolar Cku|< sns 4/23 10/27 Focai Hypertrophy No4alar HrpcrplaaU 3/23 0/2S 14/23 V, 11/27 7'27 Hepatoma 0/23 0/23 i'ZT Durtul*r HyptrpUji* i/2* 5/23 14 '27 Qjolamio-Hapatotsa 0/25 0/23 2/27 Kapatocalhilar Nero: 4/25 2/23 k '27 Coaxrol 1/23 0/23 1/23 0/23 5.23 o 1 23 STLCOPCB4029103 bd -J iSimm', li'f Nr-':. i $ STLCOPCB4029104 A&OCLC* 1240 Tabatixllofi o( Liver Lesions U R*v 889 u STLCOPCB4029105 SocriXtco Is tervs l rual AJLOCLO* 12M TsbvUtloa of IndirMlajI Livsr Lesions la Hals 0 90 Dooa Uni Vmtma Ho. rad fax Llvot Lssloas i3 I i&ji E= * wg 1_: h2 K ii a * i*r to fi :i i* o Z 3 H il ?s g 'a e i ii y9 i AI i su! I M" j ! rr i Hoi" ; I*H2*-" i| 'in- jiosior- I* !ioi- : I122T : il23" 1 : 'ur* ; , Ii2i" : ; :jo- 1 ; " v.-| 1141" i ll47" '! I 1151- i 1 ii!s- ;* : 1*4" I 131" ' * 1 :1J2- : ! Ins- ' * 1! 139- i1 i AH il*7M . i UM- '> | :l7r* ! 1 i" j , .1*4" ! V* I !tir ' I ICxtra*^ J uosr ' t20" j j 210" , 1 STLCOPCB4029106 Sacrifice Interval final (conusoed) AAOCLOft 12*0 Tabulation of Individual Liver UiImi ia Kin Dote Aniroe Level No. Senedt me i ula m >%Va ______ AH M2212230""* Z21" 224" 233" 234" 219" 230" 203" 204" i 204" 207" m0uv* i J C9* Liver Leelone a 21 1c 1-- t*av0. 0 >4 I0s> 55 *LLJ h a Li *t ii *0 1*0 H u l 3 ! * p 091 !! ii ieD i1 5! 1 1 ah: | 24124 J 233" 1 241" j 244" ! 270" i 271,: ! 272" 273" '243" ; 244" j 312" I 23 IT ! 274" ! 273" ! 294" | 297" 302" i 304" j 300" ' 311" I 314" ' 240" P P P P P P 13 STLCOPCB4029107 u. *ocL<a uu TahulMitm of Liver Leto*a > ftaii 092 i* Ciradltm Srrtmtr. * * ** *--*-* P minimal la Mvarltr > mild la sTnrry modaraca la Mvorlty markad la aa rarity Praaaat, aa grada STLCOPCB4029108 La sion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Dictular Hyperplasia ChoUngio-Hepatoma Hepatocellular Nacrosia Primary Uvar Luiou Aroclor 1260 Terminal Sacrifice Rata M If3 TO t2 Groop 3 TOI 12 > 1/25 2/25 5/25 3/23 2/20 6/23 8/27 7/24 10/27 0/25 C/25 3/25 1/23 4/20 10/23 9/27 6/24 11/27 0/25 0/25 (1/23 4/20 9/23 2/27 3/24 7/26 0/25 0/25 0/20 0/23 0/27 4/24 5/27 1/25 1/25 6/25 2/20 5/23 1/27 3/24 14/27 0/25 0/25 0/20 0/23 0/27 0/24 2/27 1/25 4/25 0/23 0/20 2/23 2/27 11/24 6/27 093 Control 12 1/23 0/24 0/23 0/24 1/23 0/24 0/23 6/24 5/23 0/24 0/23 0/24 1/23 t = First evaluation of original elide* 2 - Re-evaluation <ai original alidee 3 - Evaluation of additional tiaaue eecftioaa STLCOPCB4029109 -.<nhW SIO.T1ST Primmry lh I urtr--ii - AtOCLOOi - i?i* Terminal Iftcrtiu - lu> lU-vrabattoB af Original " linn Vacuolar Quap A-l 2/2S A-2 2/20 A-J 7/24 Focal Hyporti ophy 0/25 4/20 4/24 Nodular Hyperplasia BU* Duct Hyperplasia 0/25 1/25 4/20 2/20 1/24 1/24 ChoIaeglo-hspaSoau J/25 0/20 0/24 Htfp&tOOBA 0/25 0/20 4/24 094 Control 0/24 0/24 0/24 4/24 0/24 0/24 STLCOPCB4029110 Uiitml BIO-TIST * <. 05 . c STLCOPCB4029111 Ulmtmi tlO*T!ST i C96 TS-yv Onr>' STLCOPCB4029112 fr- Ufa**/ BI0-T1ST % 097 Ooor*r*~. STLCOPCB4029113 xxm STLCOPCB4029114 1a. 6 AAutud lO-Tl$T Ac. s n* OAe NOtTH*oo,imoii *eo*a - 239 REPOST TO MONSANTO COMPANT TWO - TEAR CHRCC.TC ORAL TOXICITY STUDY WITH AROCLOR IN ALBINO RATS H3TOPATHOLOGICAL EVALUATION or. additional liver sections MARCH 24. 197S I3T NO. 641 - 06672 STLCOPCB4029115 2:o 8 SnAutoal 810 TUT i*i **owrxo4 toAo She. NOITMiaooi.iLUNOK tM> VUrcfc *4. ms George J. LTiukM, FluD. Moousto Cornygy 800 N. Lindbergh Bird. St. Louts, Missouri 63166 . Dear Or. Unailui: ' Re: DT No. 641*06672 Histopatholog leal Erulnatioa of Additional Um Sections From Roto oi s Two - Tear Chronic Cnl Toxicity Study of Aroclor-K4. IXNT- We >n submitting herewith our laboratory report dead March 2*. 1975: prepared In connection with the above study. Very tr-olr yours. J. C. Caiaedra President STLCOPCB4029116 Wmfci./ BIO*TIST 4 241 report to MONSANTO COVCPANT TWO - TEAS CHRONIC ORAL TOXICITT STUDT WITH AROCLOR 1164' IN ALBINO RATS KISTOPATHOLOGICAL EVALUATION Or ADDITIONAL LIVER SECTIONS MARCH H, 197S 1ST NO. 661 - 0667; L Introduction Ac eha raqoaat of Dr. La-.n*k* of cha Monaanto Company. additional netiona of Uvar front a two - )r<tr chronic oral toxicity itody of Aroclor 'JYJ-- :n rata <U5T No. 611-072W) nr< procaaaad into H 6 E itainad aactioaa and avaluatad by light mieroacopy. Tha following raport praaanta tha rarulta of ihia atudy. STLCOPCB4029117 IIO.TIST 2 242 _^~4a moat liataacaii^fcp traamyrct-raUtad hiifopatlaalafleal fladiaga la tha Uvar from ra-e-ralmtioa did not diffar alfiilfVantty from that prrrrooaly raportad la Mr orLfiaal raport datad Norambar 12, 1971. Hmrmr, tbara war* thraa iwaatp Uaar tnmora datactad amopf thraa of tha animal* at tha hi{ha: traatmaot laaal (100 ppm) of tha 24-Mooth Saeriflea which wara aot pravioualy raportad. Tha othar traatmant-ralatad lasloaa raportad ara rafardad aa da- jaaaratiTa or byparplaatic la nature and they ara morpholoflc maaifeatacioa* of aa adaptdva raaponaa of tha Urar aaaociatad with hiotraaafartnation of tha taat material. la (aaaral, tha latxar Had Inga wara confined primarily to taat animal of tha final aacriflca and they arara doaa>ralatad ia Incidence aad aavanry. la coacluaioa. A roc lor MM appaara to ha a lightly tumorlfanic at lavala of 100 ppm whaa fad coadaoualy la tha diat for two yaara. Reapactfully aobalttad. DiDCSTRJAl. b:o-test LABC8ATCRIES. INC. Raport Praparad aad Reviewed by; D. C. Gordon 0. V. M.. Ph. 3. Section Ha ad. Patholofy STLCOPCB4029118 BI0-T18T IBT No. 422-0729* UrnMnta 3 243 I have anrnlMj additional ttcdou at liver frees rata oi Study MtoIht HZ-<37214 and have re -evs lusted than lot evidence at carcinogenesis. I hare inHT-- tabula tad my findings for A roc Lor Tiw on tha following pages. Tbora U evidence at a chemical affect oa tha liver which is most pronounced at time o( tha terminal (2g-Woetih) sacrifice. This consists o: a hepatocellular alto ration beginning as local hypertrophy which profresses to nodular hyperplasia, and la a lev animals to hepatoma or ehoUaftonapatoma. Tha hypertrophic calls contain large volumes oi fight staining cytoplasm which is probably rich is glycogen and endoplasmic reticulum. Is sotsa secttocs, .-.c.-s ha pad stracturaa, probably representing whorls at profiler*tire endoplasm..: reticulum, wore sees Is tha cytoplasm oi these cells. The hyperplastic nodules occupied larger areas asd often compressed surrounding cells of the normal Urer parenchyma. These nodules also contained the same hypertrophic cells. Those lesioaa srhich 1 classified as hepatomas or eholaagioheperomae were larger nodule a which showed evidence of confluence or some vnriar.oo is coll s.ne. shape, staining or a due tular or adenomatous pattern of growth, la the absence i: metaetnels, invesivenese. severe basophilia, mitoses, or other evidence oi ana plasia, I chose to call these benign tumors rather than maligna as turnon care.aomasl. There was ne evidence of metastasis or lavejiveness of theeo rumors in this study. STLCOPCB4029119 JUim+u ItO-TIST 4 244 It tbomU- bo BaTSMur fstss^\*u 4hn boidi nt athar gHfitrln *--, ifttil flmffloiitil rnilH IfT jlh***^ With xcoption al tbo ncnoUr choag** t tbo qrttfUim ot W^Wytu. tbo mar* mtm hpatoco Uolir oltoretlou woro eoaAood to aaimols of tb 24- Mosth Sacrifice. CJtAa!. R+dMi Word R. Rlcitor. D.V.M..M.S. Dipt., Am. CotUgo Vot. Pocii. STLCOPCB4029120 Primary Lira r Las iota - A roclor 124^ I 1 Month SacrUica Rats La sion Cronp 4 .cools r Chaafa rocal Kypartrophy Noc.olsr Hyparplasia ti 0/8 0/8 0/8 th z/io 0/10 0/10 tut 1/? 0/9 0/9 ' Kapatoma Ductniar Hyparplasia -holazot* Hapatoma Hapatacsllular Naerosis 0/8 0/8 0 48 1 0/10 0/10 0/10 0/10 0 49 . 1/9 0/9 0/9 Zib Control Z. 10 c - io o o ., : :: : to :n 000284 STLCOPCB4029121 Primary Lirar Lariona - Arnelor UM 8 Month Sacrifice - Rats Lation Croup TI TE TUI Vacuolar Cbanja Focal Hypertrophy 0/10 0/10 0/1 0/8 0/9 0/9 Nodular Hyparplaala 0/10 Hp*tom* 0/10 Cuerular Hyparplatia 0/10 Cholanfio-Hapatotca 0/10 Hapatocallular Neroi* t/13 0/8 0/8 0/8 0/S 0/8 0/9 0/9 3/9 0/9 0/a Control t/10 0/10 0/10 . 0/10 0/10 0/10 1/10 *\ ; + 000285 STLCOPCB4029122 7 Primary Ltni Laaioaa - Aroelor 1!M i 14 Mocth Sacrifice - *iU La lion Croup Vacuolar Chnufe Focal Hypertrophy No<uiar Hyparplaaia TI 2/10 0/10 0/10 TU 4/10 0/10 0/10 T SX 0/7 *7 1/? *1 0/7 Hepatoma - Ducrular Hyperplaaia Cholaa*ie-Hapatoma o/to 0/10 0/10 0/10 1/10 0/10 0/7 .0/7 0/7 Hepatocellular Necroaia 0/10 0/10 0/10 247 Control 1/10 0/10 0/10 0/10 . 1/10 0/19 0 '10 pv r C. w <->*-- STLCOPCB4029123 a Primary Livar La a Iona Aroclor 12M Tanninal Soerifica - Rata * 248 Laatoa Vacaolar Chanja Focal Hypartrophy No<ialar Hyparplaaia Hapatoma Duetular Hyparplaaia Choiaagio-Hapatoma Hapatocallular Neroi t 2. i 3 t ^ r Croat! I J / ? . TI hZ-+-> th 2/31 %|%y Tin VControl V ^ 1/23 ' /20 */20 *A.o ' 0/23 0/31 8/23 1/23 0/31 /l/Viy 2/:a '/lp\ 0 23 l/Jl&'Vn 3/23 Xto 5/23 0/31 f$/ 1/20 t/H 8|7ty 1/30 I 0/20 (s) Q@ 6>@> % /> > 3 3 23 1 23 X 0v h 1^ ( --^ 4 i-al *. <U.T /.i'. 000287 STLCOPCB4029124 A ROC LOR . t'tt iits?: iMUm 000288 STLCOPCB4029125 5eMflci ! Do* Animal) Lntarral j Lorol No. .j -; *od s+x i Month | C-I ! t?p= CN*'ift ! lOllM 10U" ' 1013". 1014" 1 1015" 1024F, 1027" , 1028" [ 1029" : 1030" l AHOCLOR - !.'" TahuUtion erf Io<itYirf*al Unt Uiou U JUts Ltwr L1om I'S i. LS o * - a. If H 'J II 5 f f s. z1 0 3Si E 2. = 250 T m | U9 i i i , . c9 I 9 m s j 2o31L!*S^22k ts =2--* c - -J 5Si l .c-n lottM: ' 1 Oppm 1042" 1043"; 1045" 1054F | 1057" 1 1058" j 1059" j 10 P * It-- <> V c-m 1071m! 100t>pm 1072" ; 1073" 1Q7J" i 1084F ' 1087" ' 1088" 1089" i 1090" j Control 1 301M : 802" ; 803" ' 804" 1 80 = " 1 814F ; 317" i 318" 819" . 820" ; . r -r 000289 i . STLCOPCB4029126 AROCI.OR - 124f Tabolatioa of larfividul Lirar Lioaa la ftt 151 000230 STLCOPCB4029127 A HOC LOR - UU'. TabaUtUe ai UrflvUaal Uaar Llotta ta Sat* 25 2 12 Sa critic* tnr* m 1 Dcm Aaima: Lira r L*lon* Laval No. and Su o a 2 if s| 2 L 2 1 rl * 3 5 u J| zi !l tK t* it it =t a* 24 Monti jC-I -I i `H1 SUM SU" * 59ri' /I* . 567" 569" 572" 4410 .f.. 576" 580" 588" iI1 I 589" 5 95" rt* 60ir 604" 605" 5 82" 2e '"Ai.f I, V**J" 607" 608" 609" 613" *<**+if ( */\4**lfc 621" 627" 628" H62 l" 6>t" 635" ^606'"* ^610" <620" v626" T633"* <*' e Z U* M <-> H* > * i-' b< H* 'I l*> 2o t 3s * n ie 3e I f f. k w Aj * *2 T tO., u- STLCOPCB4029128 SacrLflea if ttrvnl AHOCLOR - uml Tabulation ol Individual Llnr La riant la Vau 253 13 3om 4nima Llvar Lari--a Laval No. and Sax H -> s2 21 a3 5f 0* 1 *w f 1! M p if -8 "3 ffX. 1t 1! Si 51 ** ?* 0 f0a I *: i i 0 e is ^! ai i 3 ^ * i* z 3E *3 W 9 ~ 1u I 24 14oath c-n 65SU lOppcn 457" r*+ "! 442" W** V-3 447" 440" V4I'* c Jti*a 443 ft/iT -r-j 445"* 444 H<449"* 451" -fr 473" 44 br 4*7" ! 449" 445" 3*}j<4699S""* | 697" 700"* ^i70^"* ^707" ^704" ; 711 712" 442"* 4J3"* 491"* 702"* ft .7) II' W p I ti * ... .1. l* *n ft. J>, ---- 4h 000292 r^ STLCOPCB4029129 AfiOCLOR - t TakaUtlaa of Mirital Liror Uilu la Rata 254 14 SocrUieo latotral Do Loo* l Anlra* No. and Sox o a s ; 21 l -o:rsf. o 9 fl i* -Z o ao Urtr Lotloaa * t0 i a ; Ift! ii JTJ *4 *1 si t to ii * taf 1 *S i= i 0 I iu n il 2; >a .: ic 4 i= I~ 2 24 Monti' c-m 740U lOOppm 722" 724" 744" 746" '744" 764T Hi- ,771" 775" Ext" 733" 765" 766" 767" 776" 773" 746" 755"* 753" M-**1 Ext" If' p p p ip' if) p / p p v I Control ~*tu 11" 12" 21" ('/i 23" S3" K'-'J -4-4ul T-j 35" Ext" 46T '/i 47" 51" 52" 56" 55" 62" * I* 0* r) i-*' f # i*o 000?.?,? STLCOPCB4029130 AROCLOO Tabalatioa at Udiridual Unr Laalon* la Rats 255 * ++ +++ *+*+ * P Minimal la aararity Mild la s*rri*7 Modrata la ssa-srity Markad la sasarity PrsMat. os grads STLCOPCB4029131 MLlitonl IIO TtST JaAo*aCM*e&. At 110 FtONUOI *OAO . NOThIIOO<.IUINOi *0041 m REPORT TO MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITT STUDY WITH AROCLOR 1242 IN ALBINO RATS H3TOPATHOLOGICAL EVALUATION or ADDITIONAL LIVER SECTIONS MARCH 14. 197? IBT NO. 641 - 06671 / iT^r V'-'-f*; --A.,'' r -~Cxr' iSdArfrix..,. 0002.9;' 2 STLCOPCB4029132 MLituai BlO-TIST Jahe*io*u. %c. tTO FtOWTAOC >OAO ' NOtTH|aOO.U.UNOi *oos 1-56 George J. Leriaakas, Ph.D. UobubIs Coapwy 100 N. Lindbergh Bird. St. I.atria. Missouri (3166 Dasr Dr. Larlaskas: Rat IBT No. 641-06672 - Histopathological Eraiuatioc at Additional Lirar Sections From JLais ol a Two - Tssr Chrome Oral Toncity Study oi Aroclor 1242. We an submitting herewith our revised laboratory rsport dated March 24, 1775; prepared Is connection with the shore study. Very truly yours. J. C Calaadra Preaideni STLCOPCB4029133 157 REPORT MONSANTO COM. TWO - YEAR CHRONIC ORAL TOXICTTY rrroY with aroclor 1242 IN ALBINO RATS HETOPATHOLCGICAL EVALTATICN Or ADDITIONAL LIVER SECTIONS MARCH 29, 1975 :BT NC. 691 - 06672 -- Istrodoctioa At th* r* cm a t of Dr. LT-.:ms ol th* Momuio Cocr.parr]r. addinoral ncr.oai of Uvor ircra a toro - raar chroalc oral toxicity atody of Aroclor 1292 ;a rata iIBT No. 622-07298) tror* procoaaod ista H It E acaud aocncaa aid evaluated by light microscopy. Tb fr.llowtcg report presents thr roaulta of this ttudr. STLCOPCB4029134 \< JUJyJ BIO-T 1ST Afmhem, JU 158 IL Siimutii la noet rnniatM. the epactrua erf tree tineat-ralatod hlmnpat1nriof1l is the liver from tMe re-evaluation did sot differ (igaificantly from that previooalv reported tn oar original report datad November 12. 1971. However. there were three benign Uvar tumor detected amopg three of tba animal* at tha higheat traacmaat level (100 ppm) of tha 24-Month Sacrifice arhich vara not previously reported. Tha othar treatment-related laeionc reportad ara regardaa aa de generative or byperplaatic la nature aad they ara morphologic maai/estation* c: aa adaptive response o: tha llvar aaaociatad er-.th biotranaiermarion of the east material. In general, tha latter finding* ware 'ondlaed primarilr to t**t antma.i of tha final aacriflc* and they ware doaa-related In Incidence aad aeventy. In eonelufion. A roc lor 1242 appear* to ha tlightly rumariganic at levela of 100 ppm when fad contiaouely in tha diet for two rear*. Re*pvetfully *uhmitted. INDUSTRIAL BiC-TEST LABCRA7DRIES, INC. Raport Prepared and Reviewed by. D.E. Cordon. D.Y.M.. Ph. 3. Section Head. Pathology Raport Approved by: M. L. Kaplir^fer. PV^C. Manager. Toxtcolog^ STLCOPCB4029135 BIO-Ti JT 1ST No. 622 -07298 Monsanto Asa J *I0" I have examined additional Mctlou of liver from rata at Scad 7 Number 622-07298 and bar* re-evaluated them for evidence of carclnognnnsis. I baa* tabulated my finding! for Aroclor 1242 on the following pages. Thara is evidence of a chemical affect on the liver which Is most pronounced at time of the terminal 124-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 cholangioheoatoma. The hypertrophic cells contain large volumes of light saining cytoplasm which ;s 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 calls. The hyperplastic nodules occupied larger areas sod ofaa compressed surrounding cells of the normal liver parenchyma. These nodulee slso conatned the seme hypertrophic cells. Those lesions which I classified as hepatomas or cholaagtofcepatomse were larger nodules which showed evidence of confluence nr soma vnrisr.on :n cell line, shape, saining or a due tula r or adenomatous pattern of growth. In the absence 0: metastasis, isvasiveness, severs basophilia, mitoses, er other evidence of anapias-.a. ! chose to cal', these benign rumors rather than malignant tumors carcin omas . There was no evidence of measaaia or invasive ness of ibsse rumors t.-.ti srudy. STLCOPCB4029136 W-nW SIO-UST % 4 160 With w^Htai of tha racalar rhangimi la tfco cpoplao .< Iw^MKjtM, tha nor* oovoro hop<*nrl1iilr thon&aaa woro coateod * imtimU of tfao 24-UoBth Soaiflca. U&aJ. Word X. Richtor. D. V. U.. U. S. OipL, Am. Cotl*g Vt. Pmtiu ( a OOQrr STLCOPCB4029137 Primary Liter Laaieoa - Aroclor 1242 1 Meath Sacrifice Kata La lion Group T1 th nn Vicookr Change 0/8 2/10 1/8 Focal Hypertrophy 0/8 0/10 0/8 Nodular Hyparplaala 0/8 0/10 0/8 Hapatoma 0/8 0/10 0/8 Ooctular HyparpLaaia 0/8 0/10 1/8 Cholaagio- Hapatoma 0/8 0/10 0/8 Hepatocellular Nacroala 1/8 0/10 0/9 161 Control 2/10 o/to 0/10 0-10 C-'10 0/10 0'10 0^0301 # STLCOPCB4029138 Primary Unr Lesions Aroolor 1242 i Month Sacriilc* Rats La ton Croon t: t n im VacooUt Chang* 0/10 0/1 0/9 Focal Hypertrophy 0/10 0/8 0/9 Nodular Hyperplasia 0/10 0/1 0/9 Hspatoma 0/10 0/8 0/9 Cuctuiar Hyp*rplasla 0/10 0/8 0'9 Cho l&ag io - He p*to.*oA 0/13 0/8 z *? Hpatocllular Necrosis 1 '10 0/8 0/o 6 Control 1 ' 10 0/10 ' 0/10 o-ie 0 - 10 0'10 1. 10 00030:' ^ STLCOPCB4029139 Primary Livar Laaioa* Aroclor 1242 12 Month SocrUic* - Rata Laaioa Crop TI Ttx t m ViojoUr Chnago 2/10 4/10 o/*> Focal Hyyartrophy 0/10 0/10 1/9 Nodular Hyparplaaia 0/10 0/10 0/4 0/10 0/10 0/9 OMcnlar Hyparplaaia 0/10 1/10 0/9 C ho laaflo - Hpa toma 0/10 HaoatocalluUr Naeroaia e/io 0/10 0/10 0/9 0/10 7 163 Control 1/10 0/10 0/10 0. 10 1 .'10 0/10 0'10 OOO.^n.i STLCOPCB4029140 a Primary Llvor Laaiooa - Aroclor 12W Tarmtnal Sacriilea Riu 164 t-toa Vacuolar Chan* Toc al Hypertrophy Nodular Hyporplaaia Hepatoma Dueruiar Hyperplaala Cuolao|io-Hepatoma Hepatocellular Nee- oeie TI 7/31 i/31 0/31 0/31 3/31 0/31 1/31 Grou TII 3/30 Tin 9/20 3/30 /:o 2 /30 8/20 0/30 2/20 3/30 0/30 3/:o ` 1/23 1 30 o/:o Control ! '-3 3 23 1 23 ) :: 5 23 *J 2 2 gwr STLCOPCB4029141 7S9'\ 760"- 796F . 797 " , 79* . JOO" i . Cobtrol 36M 37" JS " 39", 40" ; 7 bT 1 77" I Vi " J 79" ! ?0" 'I - r. . . ,,*V*-Y-. STLCOPCB4029142 AROCU'R . IUZ STLCOPCB4029143 AROCLOtl - 1242 Tabulation of Individual Llvar Laalosa La Rata 11 U iU lU c-n Control lid 2" 3" 4 5" bT r. ' STLCOPCB4029144 ASOCLOK - 1342 Tabulation ot Individual Livsr Laaioas Is Rats Sac riiica In! rvil Oo> Lars l bairns No. and S*x Hfj > 0 Z !! 1 * *i o u Z1 i* i-2 ioa 0 Q. t = 24 Month |C-I ; lpym 564M 566" 59!" 567" | 56' ' 572" | 576" j 580" j 588" ; 58?" j 595" | 601T j 604" | 605" j 582" | 583" | 607" ! 608" | 609" i 613" 621" 627" 628" t 631" I 634" 635" 606" 610" 620" 626" 633" "8 io i 11 LItt L>io-- If -* 1 = 1 Si 10 * Z 168 >2 STLCOPCB4029145 Saertflc* 'Ttorral AROCLOR 1343 Tabulation of Indiridaal Unr Lotion* to Rat* 169 Do*o Anima Loxol No. and Sox s> 2 2i iS 5} IE Is M 1-* o0. Utot Lotion* *8 1 1: : i1 5-* i" z H =L e 1 4s i o ^I (* ? il c 13 2 4 Month :C-n 1 655M lOppm I 657" 542" 667" 6*0" 6*1" 6*3" 6**" 6*5" 6*6" 6*9" 651" 673" 686F 687" 689" 685" 690" 695 1 697" 700" 703" 707" 798" 711" 712" 682" 683" 691" 702" I STLCOPCB4029146 A ROC LOR - 1242 Tabulation ot latfhridaal Lirar Lalioma ia Rat* 170 u Sacrifice Intanral Oo Laral A aim* Vo. u4 Sax J 5a 1J U C4 Month C-CI 11 u TS fi. 0 >a y 74012 722" 724" 744" 746" 746" 76 ST 771" 775" Ext" 733" 765" 766" 767" 776" 773" 786" 789" 793" Zxz" * ** iiA o|tdq 1 JO 9 |IU I9 M |*9<M Urt r Lai loo* i! cr -S u 0 k 3 ii 0 *> i! ll o 0 U ]: - -= 1 5i 1" z _ 2! i4 l t* 4 V0 ** l * * *** p p p p p p *p * p 1 1 1 1 j i i i t _I *t i *: i i a 1 2 0 Oi 2c Um ?* 2v 3C !S= 31 c Il - Control ! i : i | : t I ' i ' ! <fM! U" | U" t 21-i 23- ! 33" J4"| 35-1 46F 47" 49" 51" 52 56" 59" 62" STLCOPCB4029147 ARCCLOR - 2242 Tabulation of Individual Livar Laaloat ta Rata 171 15 * * +*+ P > Minimal. la aavnnrjr Mild ia mtb rity Mod* rata la r*-ity * Markad ia aarariry < Prasaat. ao grad* STLCOPCB4029148 A&*cud aio-TMT jBrtm&m. Ac. tM H8IT*0 IOM MOTHMOO(.lUlNOi| 69*4* REPORT TO MONSANTO COMPACT TWO - TEA* CWtOfCC ORAL TOXICITY STOTT WITH aroclor m> IN ALBINO RATS K3TOPATHOLOGICAL EVALUATION or ADDITIONAL LT-'ER SECTIONS march \m 1ST NO. 641 - 06*7; 098 /' STLCOPCB4029149 JWAniW BlO-TUT Am 099 MONTAOI KOAO t NOtNUOOK.UiN04| MHI ji/ i. ms C--rg J. UvtMkM, Pk.D. MhmMd CdaptBy MH. LtodbarghBlrd. S(. Leuia. Wasson 4 JIM Dear Dr. Latrtaahaa: ' SLa: BT No. Ml-94472 HlatopatWotictl Evaluation a1 Additional Lteer Hartt--M Fran SUts af a Two * Yaar Chronic Oral Toxicity Stud-, at Arcelor 1242. . Ha art subtilfrtnfl hamiilA our ravlaod laboratory report dated March 24. 177). prepared la ccouctua the afaova study. Vary truly years, >. Xo-&*~U J. C. CaUndra President JCC/M STLCOPCB4029150 BIO-TSST 100 &XPCftT TO MOKSASTO COMPACT TWO TZAS CHXOK1C ORAL TOXtCIXT STOTT WITH AZOCU5* UG DJ AU0O1AT3 KZ3TOPATHOLOGICAL ZTAUJATiaW or AOomo*UL lttz* sxcnows MARCH 24. 1ST SO. U1 . 04472 1. lairodoctloa At tha ro^oaat of Sr. Lrrlaakaa of tfc Cottpooy, additional octloaa at Uror irun a too year chronic oral toxicity (tody o' A roc lor 1242 ta rata (OT So. 422*07291) ooro procaaaod Uta R 4 C italnod aocaoca i- t xahaatad by light microacopy. Tbo following rrport prM* tfeo roaul.a o* ttda (tody. STLCOPCB4029151 ?wd*k*l AIO-TBST Q- fanr 101 fiadiag* ta te Uver tna Rate witwhat that previetnlf wpcrtd to ear i|tMl at te ant level QOt ppeO of te H Miali we observed. Tlw eths degenerative or hyperyUefte ta arts d so adaptive reap lie ae ef the liver teat arterial. Is general, Am lease itrrfaamla d te final sacrifice and ter la conchwdna. Arecfar 1242 dam art appear to be ardaipak ta rets tod far two rears at levels sp to said tochadtog 1M ppm. BeapeettoQr oobmltud. anomiu. bio-txst laboratories. Report Prepared and Reviewed by-: -c---- D. X. Oordac., D.V.M., Ph.D. Berrien Heed. Pathology - Report Approved by: X. L. Keplhagar/WtJ-/ Tcsdeotogy "r^yryapv* '*3* STLCOPCB4029152 .Wm.w IIO-TIST J ibt xo. uz-rrm * m I Kara aaated HdHIml iiidM to iHree hern rats to BT No. U2-4TO9 arjr ftndtnga far Aroclar tttt ts Am Mowing pagaai at tut* to tbs torariml (21 HinirtO sacrtfloo. This ntfairiii to a hapatocaQolar altentfaa hogirmlwg aa fooal hypoatuvihy which yrtgtawi to nodular hyparplaata. and to a faw miamla to bspsaama ar cfaslangiehapMaaa. The hypertrophic calls contain largo roll man a to light Maturing c; inplaam which ia probably rich to glycogae and ondoplaoadc rartnal--. fat a-- aortlono. riag-obapod atrocturoa. probably loprooaaritog whorls to proUtonttra andiplaaoh reticulum. wore ma to tba cytoplasm to thorn cQa. Tba hyperplastic aodalw occupied largo- araao aaad o&on roipraaaad rurTtnavding eoQs to tho normal Uwr pat ouchfa. Tbaoo nodaloo alas rantrimod tba esma hypartrapbic caQo. Thoaa fatoioam claoatflod ao bapaaomaa or clwriongttowpatciiaaa wora larger oodulaa which ahawad orldwm to ccaflaanca or aomo rartoboa to caO Mss. shape, atatotog or a docular or adoniwiwnwia f.attorn to growth, la tho abaaaca to srtaataala. toraatraaooo. wrat basophilia. atoms, or othar ortrimwo to anaplasia, thoaa ara baaiga tamors rather thoa niHgtaiunt tvnwora (carctoaotoo). There eras ao rrldaawm to moaootaoia or iavaotraaoao to than twiwril to fhto study. tj- yps'****i .... . 090316 t- STLCOPCB4029153 STLCOPCB4029154 / hasHa Pttxmxj Um Udm AfMto ms ^iu ) uM*S**t&M Rat* Info -.-i'i Sets TX TO rm Caoeral TmwIu Cli*g* 0/8 2/10 1/0 2/10 Fseal Hyyrtfophy 0/1 0/10 0/0 0/10 Nadala* Hyparplaaia 0/8 0/10 0/0 0/10 HtfUoot 0/S 0/10 0/0 0/{0 OkkIu HrparRiAata 0/8 0/10 1/0 0/10 ClUa*i- Hapatsma 0/8 0/10 0/0 0/10 Hapatocattalar Xocrocta l/S 0/10 0/0 0/10 * ' -*v V* "' XV . 'i# .. ` , *000318 ( STLCOPCB4029155 9riataxy Urn Utima - t Meath bcttflM U*Uo TJ TO Vinalu 0/10 0/0 focal tlypcWToyhy 0/10 0/0 WoOslar Hyperplasia 0/10 0/0 Hcp&tMM 0/10 0/0 Ductalar Hyperplaiia C/10 0/0 OMlaafto KapatMaa 0/10 0/0 HepateceUalar NeeroaU 1/10 0/0 rm 9/9 9/9 9/9 0/9 0/9 0/9 0/9 0 Ccatrol 1/10 0/10 0/10 0/10 0/10 0/10 1/10 STLCOPCB4029156 -vi ~.-l .rT^nUi-.i,* .V 1 a Phaur Urar U<i( - Aradar U Maadi IictUIm &M - ' IJfcxa' TX rn < -'.t n Tiariu Ckup rKtlHyptrmpkr 2/10 0/10 4/10 o/to 0/0 1/0 M4mlr HyparyteaU - 0/10 0/10 0/10 0/14 0/0 0/0 Dwrml&r HrparpitrU 0/10 1/14 o/o- 0/10 0/10 0/0 HtpttwtUsUr !tcrta 0/10 0/10 0/10 105 Caatrol 1/10 0/10 0/10 0. 10 i/to o/:o o/n STLCOPCB4029157 Primary Laainaa - JLroclor X28I Tormina! SacrlOca Kata < 107 inis Yacnalar Oiangc T ocal Hypertrophy Notfolar Hyporplaoia Hopatoma Doetalar Hyparplaoia C bo U-a$ la - Hepatoma Hopatooallular Nacroala TI 7/11 2/11 9/11 0/31 1/31 0/11 1/31 *s TO TSX 8/14 9/20 1/19 8/20 2/10 8/20 0/10 2/23 J/10 0/10 3/20 ` 1/20 1/30 0/20 Control 1/23 0/2) 1/23 0 23 5/23 u/21 1 23 STLCOPCB4029158 T o U n its ti Id ia , fo r, AROCLOR .120 TitoUHw a Individual Unr Lttioc* lx Rat* 5*eriie Dm 6sim*t latorvsl LotoI No. kad Sox is* tiUin: 4i UYu-.h-82 3 Month C-I 11PP I S94M 594" 599 " 6 36T 637" 631 * 639" 640 " UvsrLaaloso * I. !i IS H I s *2 b8 IS z 4 r c-u; lOpptn 676M 677" 674 " 679" 640" 716T 717" 714 M 719" 7Z0 " ;c-m , 756M [lOOppca 737" , : 734" r 759" , 760" 7961* ! : 797 " I ; 794 " ; I 400" Costrol 36M 37" 34" 39" 40" 76T 77" 74" 79" SO" _1_0__8_ 9 I 1111 J *5 fc* i si l ssii it0 M 3 i" STLCOPCB4029159 STLCOPCB4029160 r-i-'-- 1 * ' - v '" ... *r *.'-r1--i**',../<*-.; -y. . .-~ \ . -v. . - ',<. _* -A -v. A . v :5h-^- ->\ '- *i:'r-'..;-_**' :" r- ' . r3':. V-;--`i-- '-J' -- II T u la n g a cta a U , lo c i! STLCOPCB4029161 STLCOPCB4029162 STLCOPCB4029163 - **. . **k-/* ; "<v<>v^i- * \> ' 'V^r > 3vv \ .3$^ itta t*o#n*T>?**;*r*- a MMA T.*,' Ilrflti aftlfcn / AJ10CL0 -1242 ' T*WUti*a 4 la^Mthial Um Laio* taJL&ts |JS 14 STLCOPCB4029164 Xr: ' vV- AIOCLOS - XM TafesUtiaa af tadlrUaal Um Lortaaa is lata Grading 3rtam * * t-t-f -*-*-* a P" Vtlatmal. la oovortry MU4 la aaToriTf Uadarata la aorariiy Utrfcsd ta sararity Ptosos*. aa pad* STLCOPCB4029165 .i - i- . ^ '? , * - V - . -' ' '\ .. .i C' Leaioo Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Luc tula r Hyperplasia C ho lan gio* Hepa toma Hepatocellular Necrosis Primary Liver UliiMi Aroeier 1242 ' 11 J Tarmlwl kcriflct Bats n i23 TO I2 Group 3 Tin 12 1 Control 12 0/31 0/27 7/31 0/31 0/27 2/31 4/30 1/19 S/30 1/30 2/19 3/30 13/20*`5/21 9/20 S/20 2/21 S/20 1/23 0/24 0/21 0/24 0/31 0/27 0/31 1/30 0/19 2/30 2/20 5/21 S/20 1/21 0/24 0/31 0/27 0/31 0/30 1/19 0/30 0*20 0/21 2/28 0/21 0/24 0/31 4/27 3/31 0/31 0/27 0/31 1/31 0/27 1/31 1/30 0/19 3/30 0/30 0/19 0/30 1/30 0/19 1.30 20 2/21 1/20 0/20 0/20 1/20 0/20 0/21 0/20 5/21 fe/24 0/21 0/24 1/21 0/24 1 - First a valuation of original slides 2 - Re-evaluation of original (Ufa* 1 - Evaluation of additional tissue Motions Four animals positive an first evaluation, not on second. a STLCOPCB4029166 BIO-TIST Primary u-nr Leaiona - AJUXXOa 1242 Terminal Satrlfico - Rats " 8U-avmhatHero al Original SHdaa - ; C-l C-2 C-J Vacuolar Change Focal Hypertrophy Nodular Hyperplasia 0/27 9/27 0/27 1/19 2/ 19 0/19 S/21 2/21 S/21 Pustular Hyperplasia 4/27 1/19 2/21 Cholanglo-hepatoma 0/27 0/19 0/21 Hepatoma 0/27 0/19 0/21 Control 0/24 0/24 C/24 6/24 0/24 0/24 nnn^^n STLCOPCB4029167 s BIO-TIST Ah*. A. tcr\rm * \ur m STLCOPCB4029168 HO-TUT 118 :*i * u urx\ct - -c : a. : \ is . z l ,1 I irjt i; | \ i ] i *1* ' n ir<M * or>or?c: STLCOPCB4029169 d SI 0 -T IS T *w.a> 1M at s yr- .r' .,-' "J& -t, uj Ji OnQ AO - .. yJ * STLCOPCB4029170 aiO-TUT Vr aA* L 120 arcni ut w >v Unh a ias r. `iarui i* wmin >TWM. jTI a w ii GnGr.?4 STLCOPCB4029171 xxm I B I T STLCOPCB4029172 AUTHORS: George J . I.e v in s k a s , Ph.D. . T IT L E : A REVIEW AND HVAI.UAT I ON OF CARCINOGENICITY STUDIES IN MI Cl: AND RATS AND MUTAGENICITY STUDIES WITH POLYCHLORINATED BIPHENYLS UIH CLXT7AL DMEH FILES Toxicology Section/St. Louis ; (COJClVJCEPTj'LQCATION) " " (' . ' ________ SPECIAL________REPORT ITYPE OF REPORT) REPORT NO.: MSL-2 00 3 JOB/PROJECT NO.: OATE: 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 (?C3s). 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. REPT. NO.: MSL-2 00 3 COPY N O : *4 ! R.1C38iBl ]i DEPOSITION | EXHIBIT I f UfV/5JSlCAil^7 I i 5'Sn-zi I SCM 019651 00033 STLCOPCB4029173  tsTR!Btmai\ COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reoorts 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, E2ND . ' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ i 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-ioa(C) <sev. j/soi sCM o 1.9652 " 000336 STLCOPCB4029174 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 ail 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 000.137 STLCOPCB4029175 cited was a paper entitled "The in vitro binding- of 2,2',5,5'-tetrachicrobiphenvl 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. Hoopingamer, 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 PC3s 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 000338 STLCOPCB4029176 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 pom of Arocior 1254 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, Koller (1977) studied mice injected with Arocior 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). Koller (1977) fed diets containing Arocior 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 Arocior were 375, 37.5, or 3.75 ppm. The only feeding regimen that was toxic was 375 ppm of Arocior 1254. Only 8 mice of that group survived for 6' months, giving a mortality of68%. 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 Arocior 1254. Koller (1977) reported that "PCBs did not produce hepatic neoplasia".There was marked liver injury in mice fed 375 ppm of Arocior 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 Arocior 1242, but this regressed and no hepatic lesions were seen 3 months after mice were returned to a control diet. Other dietary levels of Arocior 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 Arocior 1254 groups and in the group fed 375 ppm of Arocior 1242. Three months after mice were placed on a -3 SCH 019655 onorw STLCOPCB4029177 control diet, mean liver weights of each group had decreased. The decreases were highly significant for the 37.5 ppm Arocior 1254 and the 375 ppm Arocior 1242 groups. These observations are generally consistent with those of other investigators which show regression of lesions when PCB 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 laterpublications 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 laterpublication list 9/17 males and 4/17 females fedKanechior 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 STLCOPCB4029178 terminology is used to describe them, tumors were attributed only to PC3s 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 PC3s. 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, benzofcr] pyrene, butyl-nitrosourea and N-methyl-N-nitrosourea. 3 SCH 01^657 STLCOPCB4029179 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.a., 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 019653 STLCOPCB4029180 With respect to further analysis of the 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 PCB. 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 PC3s with an average chlorine content of 40% (Levinskas, 1SS1 and Weltman and Norback, 1979) and 3 on PCBs with 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 ' with those chlorine contents are not carcinogenic. Of the 3 studies with 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 Arocior 1260, the 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 hepatocellular 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 " 000343 STLCOPCB4029181 are known to be induced by carcinogens and, at the 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 STLCOPCB4029182 Sprague Dawley rats ac dietary levels of 0 and 1Q0 pom 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)". 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 hexachiorobiphenyl was more toxic than tetrachlorobiphenyl, and that the more toxic, more highly chlorinated hexachiorobiphenyl 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- SCM 019661 " 0f(W5 STLCOPCB4029183 Overall, one can surmise that 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 following' statement from a recent Surgeon General's report: "Science- and society have not yet arrived at a final consensus on the definition of a carcinogen either in the human population or in experimental animals" (DHKS, 1980). Metabolism Studies Several reviews (Goldstein, 1980; I ARC, 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 subceiluiar macromolecules. Chlorination at the 4,4' position blocks the metabolism and excretion of PCBs as illustrated below. - 10 SCM 019662 - 000346 STLCOPCB4029184 Studies in rats (Hanseil, et al. , 1977) and in mice (Morales and Macthews, 1S7S, 1979) are of interest because they report the metabolism of isomers which w-ere fed to rats for 2 years by Weianan and Norback (1979). Hanseil. et al. (1977) gave groups of male rats single intraperitoneal injections of 0.2 nmoles/kg of 2,2',5,5r-tetra- or 2,2\4,4,,5,5'hexachlorobiphenyl (TC3 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 chose 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 smooch 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. Hanseil, 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 biotransformation via the arene oxide intermediate, thereby accounting for the different slope for the " 11 " SCH 019663 . . ^ 0Q0.147 STLCOPCB4029185 elimination curve, of the latter compound." Thus, the metabolism (Hanseil, et al. L9T7) and feeding' (Weltman and Norback, 1979) study results lead co 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',5,5'-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 PC3 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, be ween 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 - SCM 019664 000.^4,3 STLCOPCB4029186 contrast co 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 Yoshimura, 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 uceri of virgin mice and fed them diets containing up to 1GQ 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 SCM 019665 0r0 STLCOPCB4029187 50 ppm.The amount of Kanechior 500 was 250 ppm or 100 ppm. Test groups consisted of 20 to 30 animais. When fed alone, only a-BHC at 250 ppm produced a high incidence of nodular hyperplasia and a moderate incidence of hepacocellular carcinoma. A diet containing 250 pom each of a-BHC and Kanechior 500 increased the number of hepatocellular carcinomas. In comparison, combinations of 100 or 50 ppm of a-BHC and 250 ppm of Kanechior 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 Kanechior 500. A mixture of 50 ppm a-BHC and 100 ppm of Kanechior 500 was without effect. Diets containing 250 ppm or 100 ppm of {3-BHC and 250 ppm of Kanechior 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 ' g-BHC and 250 ppm of Kanechior 500 or with 100 ppm of each. y-BHC and Kanechior 500 did not produce liver nodules either singly or in combination. Nagasaki, et al. (1974, 1975) reported a similar series of experiments except that Kanechior 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 Kanechior 500, they presented essentially the same data as Ito, et al. (1973a,b). While Kanechior 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 Kanechior 500. A combination of 100 ppm of a-BHC and 250 ppm of Kanechior 400 produced only a few nodular hyperplasias and 100 ppm of each in the diet did not induce any liver nodules. - 14 SCM 019666 STLCOPCB4029188 The effects of ?C3s on tumor induction by diethylnitrosamine (DEN) have been studied extensively. Male rats were given 25 ppm of DEN in their drinking water and 500 ppm of Kanechior 500 in cheir 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 Kanechior 500 only also had no tumors. When administered after DEN, Kanechior 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 corn oil solution of Kanechior 500 for 6 to 12 weeks. Kanechior 500 doses were either 0.2 ml of a 5% or 0.1 mi 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 Kanechior 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 " . . - 000.^1 STLCOPCB4029189 PCDF. The authors reported recovery of PCDF at a level comparable to 30 Mg 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 PGBs 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 STLCOPCB4029190 microsomal enzymes in Che livers of the peps. Those livers contained Kanechlor 500 residues and electron microscopy showed an increase in the smooch endoplasmic reticulum of hepatic cells. Rainbow trout were fed 6 ppb of aflatoxin (AFBx), or 100 ppm of Arocior 1254, or a combination of boch 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 Arocior 1254 alone, and growth was not affecced by 100 ppm of Arocior 1254. The number of tumor-bearing fish in the group fed AF3 L, plus Arocior 1254 was significantly reduced (to less than one-half) when compared to those fed AFBx, alone. There also were fewer tumors per liver and the tumors were smaller in chose 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-dimechylaminoazober.zene (3'-Me-DAB) or 150 ppm of N-2-fluorenylacetamide (2-FAA). Results similar to those when PC3s 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 treaced 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 -- SCM 019669 Onn.95; STLCOPCB4029191 4 months and both materials for another 2 months. No hepatocarcinomas were produced by Kanechlor 400 alone or when it was given before or during the overlap with S'-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 feE 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 phenobarbitai (S?B), in the absence and in the presence of Kanechlor 500, on the induction of rat Ever hepatoceEular carcinoma by DEN. Pretreatment of rats with DEN foUowed by DDT, by SPB, or by a combination of both produced low incidences of liver cancer whEe 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 Ever cancers. Increases resulting from joint administration of compounds after pretreatment with DEN were lower than those observed for DEN foEowed by Kanechlor 500 alone discussed earEer. 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 STLCOPCB4029192 mixture for 8 weeks. Partial hepatectcmies (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 permouse. This was applied alone as an initiator or from 5 minutes to 72 hours before initiation with 7,12-dimethylbenz[a]anthracene (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 - SCM 019671 000355 STLCOPCB4029193 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 transplantability and growth of Walter 256 carcinosarcoma in rats was inhibited byAroclor 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 intramuscular 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 (IQ3) 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 (IQ7) 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 - ?n - . SCM 019672 - 00035C STLCOPCB4029194 days, and four receiving' the higher dosage of Aroclor 1254 showed total regression of their bilateral tumors. Tumor growth rates in Aroclor-treated animals were significantly reduced. Again, body weight gain was depressed only at the 200 mg/kg dosage. An experiment was undertaken to determine if there was an optimum time period for the antitumor effect of Aroclor 1254. Intraperitoneal injections of 100 mg/kg of Aroclor 1254 were given on 5 consecutive days before tumor inoculation, on 5 days after inoculation, and daily from 5 days before until 10 days after inoculation. This study was ended 14 days after the 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 pretreatment and treatment starting with inoculation of the Walker cells. The delayed treatment starting after 5 days was least effective as ' the tumor had become established. While early treatment reduced tumor growth, it was less effective in preventing metastases and death of the animals. Kerkv-liet and Koller (1980) offered groups of male mice diets containing 10, 100, or 500 ppm of Aroclor 1254 for 15 weeks prior to injecting them with MS3, 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. - 21 SCM 019673 ' 000357 STLCOPCB4029195 Koller (1977) fed groups of mice diets containing 3.75, 37.5, or 375 ppm of Aroclor 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 intraperitoneaily with Moloney leukemia virus. None of the dietary levels of -any Aroclor afrected, 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 reports 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 00Q35r STLCOPCB4029196 possible action as tumor promoters," they added that "These compounds are being1 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 0nQ"53 STLCOPCB4029197 shorter duracion, several used 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 PCBs 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 Koller, 1978) and neither promoted.nor induced the oncogenesis of Moloney - leukemia virus in mice (Koller, 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. (1978) 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\5,5'-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 - sCH 019676 " ~ 00Q36H STLCOPCB4029198 bacterial strains W?2, TA1G0, TA93, 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 3C0 and 500, such of the preceding strains as were tested showed a negative response. The group consisting of H-L7 & M-45, WP2 try" (her* & 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 yg 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 - 00Q36 STLCOPCB4029199 Table 3, but 2,2' ,5,5'-tetr3chloro'ciphenyl (average of 4 chlorine per molecule) was presented. At 100 pg per plate, 2,2' ,5,5`-tetrachlorobiphenyl was virtually devoid of mutagenic activity. Overall, primarily because McMahon, et al. (1979) reported that 4-chiorobiphenyl 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 submitochondrial fraction of those livers was used to activate the metabolism of AFBX which was assayed for mutagenicity using ' strain TA 1538 of S. tvphimurium. 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 DNA of L-929 cells. - 26 SCM 019678 00036'- STLCOPCB4029200 Ac 1GQ (jg'/tnL. each, of che three materials caused all of the DNA to ccm.e out in fractions at the top of the gradient. The epoxide caused some breakage down to 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, the least potent, 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 mixtures were without effect on the loss of sex chromosomes used to 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. Odashina (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 PC3s 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 STLCOPCB4029201 chat 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 also have been exposed to different PCBs. Nesnow, et al. (1981) studied the cocarcinogenic action of agents which increase microsomal mixed-function oxidase activity in the C3H10T'-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 CGHlOT's 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* STLCOPCB4029202 Wong, et al. (1979) claimed, that 4-chlorobiphenyl induced an increase in unscheduled DNA synthesis in Chinese hamster ovary cell cultures in the presence of hydroxyurea, a chemical agent which suppresses normal replicative DMA 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 DMA 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 positiveresponses 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 00Q365 STLCOPCB4029203 injected with coicemide, incubated an additional 45-60 minutes, and embryos were harvested for examination. There was a high degree Gt' 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. Meddle and Bruce (L977) injected mice with Aroclor 1254 for five consecu tive days and then examined bone marrow preparations for chromosomal breakage and sperm cell 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 - SCM 019682 00Q36G STLCOPCB4029204 and epididymis from Arocior 1254 created rats were comparable to controls although, interstitial cells were increased in Arocior 1254 dosed rats. These cells showed an increase in acid phosphatase activity. The authors concluded chac the;/ "found no evidence to suggest that Arocior 1254 causes significant chromosome damage or histopathological changes in che 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 Arocior 1242. Other rats received five consecutive daily dosages of 300 mg/kg, 150 mg/kg or 75 mg/kg of Arocior 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 Arocior 1242 treated rats were prepared for cytogenetic study. Repeated dosages of 150 mg/kg and 300 mg/kg of Arocior 1254 produced decreases in the number of mitoses in bone marrow cells. Repeated dosages of 75 mg/kg of Arocior 1254 and all dosages of Arocior 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, Arocior 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 Arocior 1242 produced cytogenetic abnormalities in spermatogonial cells. The authors concluded from their studies that Arocior 1242 and Arocior 1254 did not possess mutagenic potential. Garthoff, et al. (1977) fed male rats diets containing 5, 50, or 500 ppm of Arocior 1254 for 5 weeks, after which they " 31 " SCM 019683 STLCOPCB4029205 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 spermatogonia! cells reported earlier for Aroclor 1242 (Green, et al., 1973, 1975a) did not impair the reproductive performance of male animals. - 32 SCM 019684 ore* STLCOPCB4029206 Review of Che various mutagenicity tests and their resuits prompts three comments. Chlorinaced hydrocarbons do noc generally give positive results in Salmonella strains. Therefore, the mutagenic responses with TA153S (Wyndham, ec 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 that 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 - SCH 019685 * 000361* STLCOPCB4029207 purposes, polychlorinated biphenyls should be regarded if they were carcinogenic to humans" (IARC, 1973). The other states "No conclusive evidence has thus far been reported which demonstrates that occupational exposure to PC3s 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 . ~ 000370 STLCOPCB4029208 were found are Ln 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 PC3s. 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 carcinogenicity of PC3s. "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 - SCH 019687 " _ ono;vvi STLCOPCB4029209 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 PC3s, demonstrate a pattern that is not consistent with a carcinogenic effect of PCBs. . - 36 SCM 019688 ------- STLCOPCB4029210 Footnotes L. 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. Crube, 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 PCBs 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. SCH 019689 000373 STLCOPCB4029211 R0f*ar*lrlC8S 1. Anon. (1981). The Toxicology of PC3's. An overview with 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., Steiiman, J. " (1977) PC3? and melanoma. New Engl. J. Med. 296, 108. 3. Bahn, A.K., Rosenwaike, I., Herrmann, N., Grover, P., Steiiman, J. and O'Leary, K. (1976). Melanoma after exposure to PCB's. New 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-promo ting ability of certain environmental chemicals in a two-step mouse skin tumorigenesis assay. Res. Commun. Chem. Pathol. Pharmacol. 20, 101-108. -- 5. Berry, D.L. Slaga, T.J., DiGiovanni, J., and Juchau, M.R. (1979). Studies with chlorinated dibenzo-p-dioxins, polybrominated biphenyls and polychlorinated biphenyls in a two-stage system of mouse skin tumorigenesis: potent anti-carcinogenic effects. Ann. N.Y. Acad. Sci. 320, 405-414. 6. Blazak, W.F. and Marcun, J.B. (1975). Attempts to introduce chromosomal breakage in chicken embryos with Aroclor 1242. Poultrv Sci. 54, 310-312. ` 7. Brinkman, V.A. Th. and deKok, A. (1980). Production, properties and usage. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. R.D. Kimbrough, ed. Eisavier, North-Holland, New York. pp. 1-40. 8. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. 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. 9. Cole, P. and Merletti, F. (1980). Chemical Agents and Occupational Cancer. J. Environ. Pathol. Toxicol. 3, 399-417. 10. Danz, M. and Urban, H. (1980). Elevated mitotic number in the adrenal certex as a reflection of early events in growth induction by carcinogens and promoters - a possible short-term assay. Further studies in the assessment of toxic actions. Arch. Toxicol. Suppl. 4, 19-21. 11. DHHS. Public Health Service (1980). Health Effects of Toxic Pollution: A report from the Surgeon General. Serial No. 96-15. August. U.5. Government Printing Office. SCH 019690 00Q374 STLCOPCB4029212 12. DiGiovanni. J. , Viaje, A., Berry, D.L., Slaga, T.J., and Juchau. M.R. ( 1977). Tumor-inidating- ability of 2,3,7,8-tetrachlorodiber.zop-dioxin (TCDD) and Arocior 1254 in Che two-stage system of mouse skin carcinogenesis. Bull. Environ. Contain. Toxicol., L3, 552-557. 13. Dikshich. T.S.S., Rockwood, W, , Abraham, R., Coulston, F. (1975). Effects of a polychlorinated biphenyl (Arocior 1254) on rac testis. Exp. Mol. Pathol. 22, 376-385. 14. Gaffey, W.R. (1981). The epidemiology of PCBs. Presented at the Annual Meeting of the American Chemical Society. 15. Garthoff, L.H. ,'Friedman, L. , Farber, T.M., Locke, K.K., Sobotka, T.J., Green, S., Hurley, N.E., Peters, E.L., Story, G.E., ' Moreland, F.M., Graham, C.H., Keys, J.E., Taylor, M.J., Scalera. J.V., Rothlein, J.E., Marks, E.M., Cerra, F.E., Rodi, S.B., Sporn. E.M. (1977). Biochemical and cytogenetic effects in rats caused by short-term ingestion of Arocior 1254 or Firemaster BP6. J. Toxicol. Environ. Health 3, 769-796. 16. Goldstein, J.A. (1980). Structure-activity relationships for the biochemical effects and the relationship to toxicity. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related pro ducts. R.D. Kimbrough, ed. Elsevier/North-Holland New York. pD. 151-190. * 17. Green, S., Carr, J.V., Palmer, K.A. and Oswald, E.J. (1975a). Lack of cytogenetic effects in bone marrow and spermatogonia! cells in rats treated with polychlorinated biphenyls (Aroclors 1242 and 1254). Bull. Environ. Contam. Toxicol. 13, 14-22. 18. Green, S., Palmer, K.A. and Oswald, E.J. (1973). Cytogenetic effects of the polychlorinated biphenyls (Arocior 1242) on rat bone marrow and spermatogonia! cells. Toxicol. Appl. Pharmacol. 25, 432. 19. Green, S., Sauro, F.M. and Friedman, L. (1975b). Lack of dominant lethality in rats treated with polychlorinated biphenyls (Aroclors 1242 and 1254). Food Cosmet. Toxicol. 13, 507-510. 20. HanseE, M.M., Ecobichon, D.J., Comeau, A.M. and Cameron, P.H. (1977). The relationship between retention of pure chlorobiphenyl cogeners and hepatic function in the rat. Exp. Mol. Pathol. 26, 75-84. 21. Hargraves, W.A. and AHen, J.R. (1979). The in vitro binding of 2,2',5,5'-etrachlorobiphenyl metaboEtes to rat Ever microsomal pro teins. Res. Comm. Chem. Pathol. Pharmacol. 25, 33-52. 22. Heddle, J.A. and Bruce, W.R. (1977). Comparison of tests for mutagenicity or carcinogenicity using assays for sperm abnormaEties, formation of micronuclei, and mutations in Salmonella. In: Origins of Human Cancer. Cold Spring Harbor Conferences on CeE Proliferation. H.H. Hiatt, J.D. Watson and J.A. Winsten, eds. Cold Spring Harbor Laboratory. Vol. 4 Book C, pp. 1549-1557. - 39 SCM 019691 nnor?^ STLCOPCB4029213 23. Headricks, J.D., Putnam. T.?.. Bills, D.B., and Sinnhuber, R.O. (1977) Inhibitory effect of a polychlorinated biphenyl (Aroclor 1254) on Aflatcxm 31 carcinogenesis in rainbow trout (Salmo .gairdnerl). J. Natl. Cancer Inst. 59, 1545-1551. Hoopingarner, R., Samuel. A., and Krause, D. (1972). Polychlorinated biphenyl interactions with tissue culture cells. Environ. Health Perspect. _1, 155-153. 25. Hsia, M.T.S., Lin, F.S.D. and Allen, J.R. (1978). Comparative mutagenicity and toxic effects of 2,5.2'5;-tetrachlorobiphenyl and its metabolites in bacterial and mammalian test systems. Res. Comm. Chem. Pathol. Pharmacol. 21, 485-496. 26. IARC (1978). Polychlorinated biphenyls and polybrominated biphenyls. [ARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. Volume 18, 43-103. IARC. Lyon, France. 27. Ito, N., Nagasaki, H., and Arai, M. (1973a). Interactions of liver tumorigenesis in mice treated with technical polychlorinated biphenyls (PCBs) and benzene hexachloride (BHC). In: New methods in environmental chemistry and toxicology. F. Coulston, F. Korte and M. Goto, eds. Int. Acad. Print. Co., Ltd. Tokyo, pp. 141-147. 23. Ito, N., Nagasaki, H., Arai, M. , Makiura, S., Sugihara, S. and Hirao, K. (1973b). Histopathologic studies on liver tumorigenesis induced in mice by technical polychlorinated biphenyls and its promoting effect on liver tumors induced by benzene hexachloride. J. Natl. Cancer Inst. 5_L, 1637-1646. 29. Ito. N., Nagasaki, H., Makiura. S. and Arai, M. (1974). Histopathological studies on liver tumorigenesis in rats treated with poly chlorinated biphenyls. Gann 65. 545-549. 30. Ito, N., Tatematsu, M. , Hirsoe, M. Nakanishi, K. and Murasaki, G. (1978). Enchancing effects of chemicals on production of hyperplastic liver nodules induced by N-2-fluorenyiacetamide in hepatectomized rats. Gann 69, 143-144. 31. 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. 32. Kerkvliet, N.I. and Kimeldorf, D.J. (1977a). Inhibition of tumor growth in rats by feeding a polychlorinated biphenyl, Aroclor 1254. Bull. Environ. Contain. Toxicol. 1_8 243-246. 33. Kerkvliet, N.I. and Kimeldorf, D.J. (1977b). Antitumor activity of a polychlorinated biphenyl mixture, Aroclor 1254, in rats inoculated with Walker 256 carcinosarcoma cells. J. Natl. Cancer Inst. 59, 951-955. SCM 019692 00Q.T7G STLCOPCB4029214 34. Kerkviiet, I. and Roller, L.D. (1980). Effect of cadmium, arsenic and PCB's on tumor-directed cell-mediated cytotoxic immune reactions in mice injected with MSB sarcoma ceils. In: Inadvertent Modification of che Immune Response. Proceedings of the Fourth FDA Science Symposium. U.S. Naval Academy, August 28-30, 1978. HH5 Publ. FDA-80-1074, pp. 275-279. 35. Kimbrough', R.D. (1979). The carcinogenic and other chronic effects - of persistent halogenaced organic compounds. Ann. N.Y. Acad. Sci. 320, 415-418. 36. Kimbrough, R.D. (1980). Occupational exposure. In: Halogenaced biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. R.D. Kimbrough, ed. Elsevier/North-Holland, New York pp. 373-397. 37. Kimbrough, R., Buckley, J. , Fishbein, L., Flamm, G., Kasza, L., Marcus, W., Shibko, S. and Teske, R. (1978). Animal Toxicology. Environ. Health Perspect. 24, 173-184. 38. Kimbrough, R.D. and Linder, R.E. (1974). Induction of adenofibrosis and hepatomas of the liver of BALB/cJ mice by polychlorinated bi phenyls (Aroclor 1254). J. Natl. Cancer Inst. 53, 547-552. 39. 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. 40. Kimura, N. and Baba, T. (1973). Neoplastic changes in the rat liver induced by polychlorinated biphenyl. Gann 64, 105-108. 41. Kimura, N.T., Kanematsu, T. , and Baba, T. (1976). Polychlorinated biphenyl(s) as a promoter in experimental hepatocarcinogenesis in rats. Z. Krebsforsch. Klin. Onkol. 87, 257-266. 42. Koiler, L.D. (1977). Enhanced polychlorinated biphenyl lesions in Moloney leukemia virus-infected mice. Clin. Toxicol. 11, 107-116. 43. Levinskas, G.J. (1981). Toxicity of Aroclor products 1242, 1254 and 1260 to the liver of albino rats. 44. Lichti, U., Yuspa, S.H. and Hennings, H. (1978). Ornithine and S-adenosyimethionine decarboxylases in mouse epidermal cell cultures treated with tumor promoters. In: Carcinogenesis, Vol. 2 Mechanisms of tumor promotion and cocarcinogenesis. T.J. Slaga, A. Sivak, and R.K. Boutwell, eds. Raven Press, N.Y. pp. 221-232. 45. Makuira, S., Aoe, H., Sugihara, S., Hirao, K., Aral, M. and Ito, N. (1974). Inhibitory effect of polychlorinated biphenyls on liver tumorigenesis in rats treated with 3'-methyl-4-dimethylaminoazobenzene, N-2-fluorenylacetamide, and diethylnitrosamine. J. Natl. Cancer Inst., 53, 1253-1257. - 41 SCH 019693 000377 STLCOPCB4029215 46. Matthews. H.B. and Kaco, S. (L979). The metabolism and disposition or haloge.naced aromatics. Ann. N.Y. Acad. Sci. 320, 131-137. 47. McMahon. R.E., Cline, J.C. and Thompson, C.Z. (1979). Assay of 355 test chemicals in ten tester strains using a new modification of Che Ames cesc for bacterial mutagens. Cancer Res. 39, 632-693. 43. Morales, N.M. and Matthews. H.B. (1973). In vivo binding of PCBs " to hepacic micromolecules in mice. Toxicol. Appl. Pharmacol. 45, 334. 49. Morales, N.M. and Matthews. H.B. (1979). In vivo binding of 2,3.6,2,',3' ,6'-hexachlorobiphenyl and 2,4.5,2',4',5`-hexachloro- biphenyi to mouse liver macromolecules. Chem-3ioi. Interact. 27, 99-110. ~~ 50. Nagasaki, H., Tomii, S., Mega, T., Marugami, M. and Ito, N. (1972). Hepacocarcinogenicicy of polychlorinated biphenyls in mice. Gann 63, 805. 51. Nagasaki, H., Tomii, S., Mega, T. Sugihara. S., Mivata, Y. and Ito, N. (1974). Analysis of various factors on liver carcinogenesis in mice induced by benzene hexachloride (BHC) and technical polychlorinated biphenyls (PCB's). J. Nara. Med. Assoc. 25 : 635-648. 52. Nagasaki, H. , Tomii, S., and Tsumashika, Y. (1975). On a'main cause for the formation of hepatic tumors of mice by BHC and PCBs. Nippon Eiseigaku Zasshi 30, 134. (Translation by Ralph McElroy Co., Custom Division, Texas). 53. NCI (1978). Bioassay of Aroclor 1254 for possible carcinogenicity. NCI-CG-TR-38, DHEW. Publication No. (NIH) 73-838, Washington, D.C., U.S. Department of Health, Education and Welfare. 54. Nesnow, S., Leavitt, S., Garland, H., Vaughan, T.O., Hyatt, B., Montgomery, L., and Cudak. C. (1981). Identification of cocarcinogens and their potential mechanisms of action using CSHIOT^CLS mouse embryo fibroblasts. Cancer Res. 41, 3071-3076. 55. Nilsson, B. and Ramel, C. (1974). Genetic tests on drosophila melar.ogaster with polychlorinated biphenyls (PC3). Hereditas 77, 319-322. 56. NIOSH (1977). Criteria for a recommended standard. Occupational exposure to polychlorinated biphenyls (PCBs). DHEW (NIOSH) Publication No. 77-255, Washington, D.C., U.S. Government Printing Office. 57. Nishizumi, M. (1976). Enhancement of diethylnitrosamine hepatocarcinogenesis in rats by exposure to polychlorinated biphenyls or phenobarbital. Cancer Lett. , 2, 11-16. 58. Nishizumi, M. (1979a). Effect of DDT and phenobarbital on the accelerating action of PCBs in liver cancer generation due to diethylnitrosamine. Nippon Eiseigaku Zasshi (Jpn. J. Hyg.) 34, 197. (Translation for EPA by Literature Research Company). - 42 SCM 019694 ooor.78 STLCOPCB4029216 59 Nishizumi, M. (1979b)-. Effect of phenobarbical, dichlorodiphenvltrichioroethane, and polychlorinated biphenyls on diethylnitrosamine- induced hepatocarcinogenesis. Gann 70, 335-337. 60 Nishizumi, M. (1930). Reduction of diethylnitrosamine-induced hepatoma in rats exposed to polychlorinated biphenyls through their dams. Gann 7_1, 910-912. 61 Norback, D.H., Reddy, G. and Cihla, H.P. (1980). Oncogenic, cytotoxic and enzymatic effects of PCBs on COHIOT^ cells. ACS, Ann. Mtg. , San Francisco, Abstract no. 153 in section on Environment. 62 Norback, D.H. and Swedo, G.J. (1979). Transformation in vitro induced by Aroclor 1254: Ultrastructure of transformed cells. Toxicol. Appl. Pharmacol. 48, A182. 63 Norback, D.H., Weltman, R.H., and Cihla, H.P. (1981). Malignant transformation of CSHlOT's cells by polychlorinated biphenyls. Fed. Proc. 40, 757. 64 Odashima, S. (1976). The cooperative development in Japan of methods for screening chemicals for carcinogenicity. In: Screening tests in Chemical Carcinogenesis, International Agency for Research on Cancer. R. Motesano, M. Bartsch and L. Tomatis, eds. IARC Scientific Publ. No. 12, Lyons, France, pp. 61-75. 65 Peakall, D.B., Lincer, J.L. and Bloom, S.E. (1972). Embryonic ' mortality and chromosomal alterations caused by Aroclor 1254 in ring doves. Environ. Health Perspect. _1, 103-104. 66 Peraino, C., Fry, R.J.M., and Grube, D.D. (1978). Drug-induced enhancement of hepatic tumorigenesis. In: Carcinogenesis, Vol. 2. Mechanisms of Tumor promotion and Cocarcinogenesis. T.J. Slaga, A. Sivak, and R.K. Boutwell, eds. Raven Press, N.Y. pp. 421-432. 67 Pienta, R.J. (1980). Transformation of Syrian hamster embryo cells by diverse chemicals and correlation with their reported carcinogenic . and mutagenic activities. In: Chemical Mutagens: Principles and Methods for Their Detection Vol. 6. F.J. deSerres and A. Hollaender, eds. Plenum Press, New York. pp. 175-202. 68 Pitot, H.C., Barsness, L. and Kitagawa, T. (1978). Stages in the process of hepatocarcinogenesis in rat liver. In: Carcinogenesis, Vol. 2. Mechanisms of Tumor Promotion and Cocarcinogenesis, T.J. Slaga, A. Sivak, and R.K. Boutwell, eds. Raven Press, N.Y. pp. 433-442. 69 Preston, B.D., Van Miller, J.P., Moore, R.W. and Allen, J.R. (1981). Promoting effects of polychlorinated biphenyls (Aroclor 1254) and polychlorinated dibenzofuran-free Aroclor 1254 on diethylnitrosamine- induced tumorigenesis in the rat. J. Natl. Cancer Inst. 66, 509-515. 70 Probst, G.S., McMahon, R.E., Hill, L.E., Thompson, C.Z., Epp, J.K. and Neal, S.B. (1981). Chemically-induced unscheduled DNA synthesis in primary rat hepatocyte cultures. Environ. Mutag. 3, 11-32. - 43 SCM 019695 " Ono STLCOPCB4029217 71. RibeLin. W.E. (1931). Personal communication. 72. RLnkus. 5.J. and Legator, M.S. (1930). The need for both in vitro and in vivo syscems in mutagenicity screening. In: Chemical Mutagens: Principies and Methods for their detection. Vol. 6. F.J. deSerres and A. HoLIaender, eds. Plenum Press, Mew York. pp. 365-473. 73. Roberts, J.R., Rodgers, D.W., Bailey, J.R. and Rorke, M.A. - (1973). Polychlorinated biphenyls: Biological criteria for assessment of their effects on environmental quality. National Research council of Canada, Publication no. NRCC 16077. 74. Safe, S. (1980). Metabolism, uptake, storage and bioaccumulation. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. R.D. Kimbrough, ed., Elsevier/North-Holland New York. pp. 81-107. 75. Squire, R.A. and Levitt, M.H. (1975). Report of a workshop on classification of specific hepatocellular lesions in rats. Cancer Res. 35, 3214-3223. ' 76. Stadnicki, S., Lin, F.S.D. and Allen, J.R. (1979). DNA single strand breaks caused by 2,2',5',5-tetrachlorobiphenyl and its metabolites. Res. Comm. Chem. Pathol. Pharmacol. 24, 313-327. 77. Stott, W.T. and Sinnhuber, RiO. (1978). Trout hepatic enzyme activation of aflatoxin B], in a mutagen assay system and the - inhibitory effects of PCB. Bull. Environ. Contam. Toxicol. 19, 35-41. 78. Sugimura, T. , Kawachi, T., Matsushima, T., Nagao, M., Sato, S. and Yahagi, T. (1977). A critical review of submammalian systems for mutagen detection. In: Progress in genetic toxicology. D. Scott, A.B. Bridges, and F.H. Sobels, eds. Elsevier/North-Holland Biomedical Press, pp. 125-140. 79. Sugimura, T., Sato, S., Nagao, M., Yahagi, T., Matsushima, T., Seino, Y., Takeuchi, M. , and Kawachi, T. (1976). Overlapping of carcinogens and mutagens. In: Fundamentals in Cancer Prevention. P.N. Magee, S. Takayama, T. Sugimura and T. Matsushima, eds. Univ. of Tokyo Press, Tokyo/Univ. Park Press, Baltimore, pp. 191-215. 80. Tazima.Y. (1980). Chemical mutagenesis in the silkworm. In: Chemical Mutagens: Principles and Methods for their Detection. Vol. 6, F.J. deSerres and A. Hollaender, eds. Plenum Press, N.Y. pp. 203-23S. 81. Turusov, V.S. and Takayama, S. (1979). Tumours of the liver. In: Pathology of Tumours in Laboratory Animals. International Agency for Research on Cancer. World Health Organization. Lyon France. V.S. Turusov, Editor-in-Chief. Vol. II-Tumours of the mouse, pp. 193-233. 82. Uchiyama, M. and Chiba, T. (1974). Co-carcinogenic effect of DDT and PCB feedings on methylcholanthrene-induced chemical carcinogenesis. Bull. Environ. Contam. Toxicol. 12, 687-693. - 44 - SCM 019696 ' 000380 STLCOPCB4029218 83. Wasserman, D., Miller, and Wasserman, M. (1978). . Polychlorinated biphenyl-induced rat liver adenomas. Toxicol. Eur. Res. 1. 159-172. 84. Weisburger, J.H. and Williams, G.M. (1980). Chemical Carcinogens. In: Casarett and Doull's Toxicology, J. Doull, C.D. Klaasen, and M.O. Amdur, eds. , Macmillan Publishing Co., Inc., New York, pp. 84-138. 85. Weltman, R.H. and Norback, D.H. (1979). Hepatic ultrastructural changes of rats exposed to polychlorinated biphenyl isomers: Hexachlorobiphenyl-induced neoplastic nodules. Toxicol. Appl. Pharmacol. 48, A181. 86. Wong, A., Basrur, P.K. and Safe, S. (1979). The metabolically mediated DNA damage and subsequent repair by 4-chlorobiphenyl in Chinese hamster ovary cells. Res. Commun. Chem. Pathol. Pharmacol. 24, 543-550. 87. Wyndham, C. 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 MOci, 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. - 45 SCM 019697 STLCOPCB4029219 TAIII-E 1 PCBsj_Tabu 1.11 i on of Liver Nodules Reported in Mice1 Approx. wt. X Cl2 Product Strain Dosing Duration pen in diet (ppm) No. Anima1s Nodular Hyperplasia Hepatoma Hepato cellular Carcinoma Keference 52-54 Aroclor 1254 DAI.B/cJ 6 mos3 11 mos 0 300 0 300 34 24 24 22 - - " 0 Kimbrough 1 - +. l,iruler (1974) 0- 9< - i Kanechlor 500 dd 32 weeks 0 100 250 500 6 12. 12 12 0 0 0 7 - (-)5 0 Nagasaki, et a 1. 0 (1972) 0 I to, et a 1. ( 19 73a,b) 5s Kanechlor 500 dd 32 weeks 0 100 250 500 20 18 20 17 - 9 - 0 Nagasak i , et a 1 . - 0 (1974, 1975) -0 - 76 Kanechlor 500 dd7 32 weeks 0 100 250 500 12 19 20 17 - - 4 - 0 Nagasaki , et a 1 . - 0 (1974, 1975) -0 - 0a oo STLCOPCB4029220 48 IA O X o 3 o r--* O %0 *3 O' >o CD '0 ! Kanechlor 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 a 1 . 0 (1972) 0 Ito, et al. (1973a.b) 0 0 Nagasaki, et a 1. 0 (1974, 1975) 0 0 0 Nagasaki, cl a 1. 0 0974, 1975) , 0 0 TABLE 1-continued lTdlsj_Taln^hil ion pi' l.iver Nodules Reported in Mice1 Approx, wt. % Cl12 3P4r*o*duct Strain Dosing Duration pen in d i et (ppm) No. Anima1s 40-42 Kanechlor 300 dd 32 weeks 0 100 230 500 6 12 12 12 Kanechlor 300 dd 32 weeks 0 100 250 500 20 19 19 20 Nodular Hyperplasia o' 0 0 0 - - HepaLoma _ - - - " Hepato cellular Carcinoma 0 0 0 0 0 0 0 0 lie 1 e reure Nagasaki, cL a 1. (19/2) llo, cL al . (19/ !!a , h ) * Nagasaki , et a 1 . (1974, 1975) Kanechlor 300 1.17 32 weeks 0 100 250 500 12 19 20 20 - - - 0 Nagasaki , et a 1. - 0 (1974, 1975) -0 -0 1 Hales, except as noted. 2 From Brinkman and deKok (1980). 3 Held an additional 5 months before sacrifice. 4 Ho. of animals. One had 2 hepatomas for a tumor total of 10. 3 Apparently same data reported as hepatoma and hepatocellular carcinoma. See references and text. 6 Apparently same data reported as nodular hyperplasia and hepacellular carcinoma or Lumor. See references and text. 7 Females. t/> n 2 o o o tO-- o oo o *0 t -fl w S s* I 333 i iii lilt 33 3 3 3 3 *>X AI CCi 3-iii -- :i _O3=. "*551i1 o -j' ---vf iii till 033-- 33--: ili tii 5 31 35 34 1 O r*. o i O .""I l I 3 ft 33 3 --3 3 3 SCM 019700 ooo;is4 STLCOPCB4029222 STLCOPCB4029223 -23 3MH X3 ---- 8 - 3 *r ^ 3 3 - 3 "3 5 -j Z3 w. 3 s -j 32 --i -- oV- s- =-< 3:3 -< 3 c3 s3 r5 a - 3 03 33 33 "3 XX 32 U _I * _* 2- W3 -3 vT x wu OX- ---*aU3 uu X. Q a f- XX SCM 019702 OOOaBG STLCOPCB4029224 73 7 - .7. * 37 5 ju *1 1U -ns -s s = CW cw 3: 3 3 -- - v:i --~aw'w --.s3 --jW --jg: w--u jwIct US3W S3J '7w u0 7U 7 3> H VI _* > 37 *74 431 wa- 3 -* ZWl a A'33-a.. --w 7 < (<W --W* S3+m 7 31 < 4 <34 T <2_ -S -- -- SCM 019703 0H0:<67 STLCOPCB4029225 Product Aruclor 1242 Aroclor 1221 Mode Tine TAIJj.E_3-co.il inued sywiAUYOK !il.!^KLl!^Q^NKS!s_STuiits_witil i-riiu Ollier Treatment1 Hode Species Ohscrval ions Dirt Diet It.dure Be lore Moloney 1 cukeiu i d v i r ii Mo 1uncy 1 cukemi a vi rus 1 III rapei 1 1 UIIL'4 1 llalb/c wuosc lot 1 4peiilouea1 Itj 1 h/c mouse Did not .1 led oncogenesis oi Mo 1ouey I eukumi a vi ik. Did nut allecl oncogene*I * o 1 Mo 1uncy leukemia virus, He I . Ko1ler, 1*1/7 Ku1t e r, 1')!) 'Chcoicdls used: 20-McCh 20-mLhylcho Wnihrcne Ci-BIIC * a-benzene hexachloride p-BIIC * fl-bciuene hesschtoride DEN * dielhyInilrossainc AKB| s nUatuMin Bt 2~}`AA N-2-fluorenyldCctaide 3*-Me-DAH s 3*-acthyI -4-diaet hy 1 aminoazobenaciie / UDT = dichlorodipheny IL richinruelbane SPB = sodium pbenubarbitdl 7 , 12'DBA * 7,2-dimct by Ibriu |n |ant bratrue TPA - 12"0'letradecanoyIjdiuibuI - 13-afelale ``Pregnant damn dosed with Kanuchlor 400. UJJsprtng dosed with DEN. Established tissue culture ceil line derived from Moloney sarcoma virus-induced tumors in |i6 mice. SCM Tester Strain Product AKOCI.OK 1268 AHOCI.UK I25A AK0CL0K AK0CI.UK 1251 Kaiicchlor SOO2 Kanechlor SOU C3076 D3052 Neg. Neg. G46 TAOH Neg. Keg. Keg. Neg. IMIlKJi H'Hs _Ti*l**ilat ion oi Hi t'rob) a J_Hul agi*ni l*i ly Tcsls s:. >yi*! ifliur I tux I'AIOO TAI0OII TAI535 1'A 1536 TAI537 TAI53B Neg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. l*o 1 Neg. Neg. Neg. A Neg. Neg. k CO I I WP2 W1`2 uvrA* He 1 . Neg. Nug. Neg. Wyinlli.ini , el .1 l . 1 *1 / O l*i tihsl , el .ij_. |`JHI lleddtc ami liiucc I'H/ Wyiulh jut, el u t . l`J /b Oil j bh l ui j Sug nun r u , el ul. I'J/e 2,2*,5,5-letraclilorobipheny l A Neg. Wyiulli jui , el. al. I'i/I. a^'S.S'-leiraclilorolit|il>enyl Keg* Neg' Neg' Neg' llsia, el ill. i'j;b Kanechlor 300x a Neg. Neg. Nfcg. Neg. 0| jbh i uta I'J )b Kanechlor 300 if AKOCLOH 1221 Neg. Neg. . A AA I'uu' Neg. Sugiumra el j 1 . 1 >l/i. Wymlluiu, el jl. I'1/g 4-chlorobiphtfny1 A* Hu.' Wyndluiu, el j|. I'l/i. SCM 0 1 9 7 0 5 - 4-ehlorobiphenyl He*. Neg. Neg. Neg. Neg. 1. All lest* done with amJ wi(houl icruhunu 1 activation except as notc1 Neg. Neg. Neg. Neg . Neg. hi H.ilmn . el .. 1. fl/'l. 2. Ollier strains tested, tee lent ami reference. s Hoi tibled. Neg. = Negative. NegU - Only I cs 11*<| ui I lum i jilivutiun. Neg* s Only tested with jilivjlnm. ii-" ID l'b' s I'libilivt* only willi .m I iv.ii i>iii. STLCOPCB4029228 ri decision Analysis. Phil 's _eroy, A. A.. Gaylord, rger. T. 1982. Reducing nduced hospiuiizinoa: uoliziboo review. Drug > 986. Pharmaceutical inenenc drug competition "facts of the Drug Price 4 Puent Terra Restora14. Pharm. Med. 1:177- rolley. P. D.. Brown. T. usubstitutioo law cooutioo? A/vt. Intent. Med. 1986. Med. Lett. 28:1-2 Ann Rev Phurmacoi Toxicol. I9Q7 27 87-ltt 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|2Hio-*C1,,, with n = 1--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 (1) 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 dichioro-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 and to any copyright covering this paper. STLCOPCB4029229 STLCOPCB4029230 .o o o 88 KIMBROUGH were produced (Aroclor 1242 and 1016). In 1977 the company ceased produc tion entirely (2). Many PCBs manufactured in the past (3) are still in use in old transformers, but even this use is decreasing. The estimated cumulative production and consumption of PCBs in the United States 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 lipophilicity, they have accumulated in the environment and in organisms. They have been identified in indoor air (4) at concentrations of 0.1 /tig/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 centrations in soil (0.01 mg/kg-100 mg/kg) (10). They do not occur naturally. Thus, their presence in the environment is linked with human activities, and concentrations are higher in urban and heavily industrialized areas than in rural and remote areas. However, trace amounts are also found in remote areas, since the air may transport such chemicals over large distances. Although PCBs are no longer used commercially in the United States because of their persistence, they are still present in our environment. A number of transformers and capacitors that contain PCBs, however, are still in use. Results of laboratory experiments showed that pyrolysis of PCBs at temperatures of 200-600C could result in the formation of significant amounts of the more toxic polychlorinated dibenzofurans (PCDFs) (II). In February 1981, an electrical fire occurred in a New York Stale office building in Binghamton. N.Y. The fire, which originated in a switch gear in the basement, caused the bushings to crack on a nearby transformer. About 180 gallons of PCB dielectric fluid Pyralon (65% Aroclor 1254. 35% chlorin ated benzenes and trace additives) were lost. A fine layer of oily soot covered many of the internal surfaces of the 18 floors of the building. Analysis of a soot sample showed that it contained various isomers of chlorinated di benzofurans. 2.3.7.8-tetrachlorodibenzodioxin, other chlorinated dibenzodioxins, and chlorinated biphenylcnes. Some of these chemicals are much more toxic than the PCBs. Because of these findings the Binghamton state office building was closed, and workers wearing respirators and pro tective clothing began an extensive cleanup of the building. The cleanup operations lasted four years, and the cost has been enormous (12). Since then several other transformer fires have occurred. These fires have not resulted in as much contamination as the one in Binghamton, partly because the transformers in the other fires were usually located in a separate vault (13). The problems with polybrominated biphenyls (PBBs), which are also a mixture of chemicals, have been quite different. In 1970 a chemical company in Michigan manufactured polybrominated biphenyls as flame retardants. The same company also produced magnesium oxide, a chemical commonly mixed HUMAN HEALTH EFFECTS OF PCBs <fc PBBs 89 into feed for livestock. The flame retardant was called Fircmaster and the magnesium oxide, Nulrimastcr. In 1973 some bags of Fircmaster were accidentally sold as Nulrimastcr 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 the exposure little was known about the 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. HUMAN EXPOSURE Because PCBs are ubiquitous and very persistent in the environment, humans have been and will continue to be exposed to them, particularly in in dustrialized countries. PCBs may be inhaled in small amounts through the air or ingested through food. In the United States today, people are primarily exposed to PCBs by consuming fish from contaminated waters (9). In the past, some farm families were exposed to PCBs from dairy products; these PCBs originated from coating material used in the inside of silos (15). In addition, workers who repair transformers and workers who handle toxic wastes may also be exposed (16). The PCB products that were manufactured by Monsanto in the United Stales had the trade name "Aroclor." The particular kind of Aroclor is identified by a four-digit number. The first two digits refer to the 12 carbon atoms, and the second two refer to the percent, by weight, of chlorine in the mixture. Thus, Aroclor 1254 contains about 54% chlorine, and Aroclor 1260, about 60% chlorine. The composition of this mixture of chemicals, with different properties, changes once it gets into the environment and into organisms. Some com ponents of the mixture are more easily degraded in the environment than others. As a result the PCBs identified in the environment resemble Aroclor 1254 but are not identical to it. Similarly, the PCB mixtures found in humans usually resemble Aroclor 1254 if exposure occurred primarily through the environment. A different composition of the PCBs may be found in serum or adipose tissue samples from occupationally exposed workers. For instance, if the workers are primarily exposed to Aroclor 1016 or Aroclor 1242, which contain much less of the more highly chlorinated homologs. then their gas-chromatographic patterns resemble a combination of Aroclor 1016 or 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 humuns can be used to determine whether the exposure occurred primarily 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 9.1% of the US population eats fish, the average annual per capita consumption is small: 15 lbs. per year (6, 7). If PCBs arc to be quantitated in fish, the edible portion rather than the whole fish must be examined. Because of their lipophilicity, PCBs are preferentially stored in the hepalopancreas 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 are 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 2.6-4.5% (21). At 2% fat, I liter (I) of milk would contain 0.04 mg or 40 Mg if the PCBs were present in milk fat at a concentration of I ppm. If an infant weighed 5 kg and imbibed 750 ml of milk per day. it would take in about 6 Mg/kg. * dose that exceeds the 1.5 Mgfkg dose calculated as acceptable by Cordle cl al (6). At 1% milk fat this dose would be reduced to 3 Mg^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 time. However, at 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 cat fish. The con centrations at which such exposure presents a risk are not clear. Recently, Cordle el 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 46.5 mg of PCBs per year; this amount ranged from I4.l7lt 114.31 mg/year/person. The calculated mean daily dose received by the exposed group was 1.7 Mgdtg/day and ranged from 0.09 to 3.94 Mg^kg/ day. Thus, the average sports 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 does 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- STLCOPCB4029232 ob o CO 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 lo PCBs was not clear. 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 Stales, and in other countries to elucidate the toxic effects of PCBs. These data arc summarized in several detailed reviews (16, 25, 26). Animal Studies This article addresses primarily the human health effects of PCBs and PIIBs. 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 lo the human situation la difficult. Some animal species, such as 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 lo 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 Aroclor 1254 affects reproduction in rats at much lower doses than does Aroclor 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 r HUMAN HEALTH EFFECTS OF PCBs & PBBs 93 a very small portion of the mixture, particularly those with less chlorine by weight, such as Aroclor 1242 or Aroclor 1016. Recently, Schaeffer el al (33) found that a German PCB mixture--Clophcn A-30. with an average composi tion of 1% monochlorobiphenyl, 20.7% dichlorobiphenyl, 57.4% trichlorobiphenyl, 17.3% telrachlorobiphenyl, 1.8% pcntachlorobiphenyl, 1.0% hexachlorobiphenyl, 0.6% heptachlorpbiphenyl, and 0.1% octachlorobiphenyl--produced a 3% incidence of hepatocellular carcinoma, whereas Clophcn A-60 produced a 61% incidence of hepatocellular carcinoma in Wislar rats. The incidence of the disease in the controls was 2%. The Clophcn A-60 had an average composition of 0.2% monochlorobiphenyl, 1.1% di chlorobiphenyl, 2.2% trichlorobiphenyl, 3.1% telrachlorobiphenyl. 19.8% pcntachlorobiphenyl, 43.2% hexachlorobiphenyl, 25.3% heptachlorobiphenyl, 4.7% octachlorobiphenyl, and 0.3% nonachlorobiphenyl. Sim ilarly, Norback A. Wellman (34) and Kimbrough et al (35) were able to produce hepatocellular carcinomas in rats with Aroclor 1260, the more highly chlorinated Monsanto product. When Aroclor 1254 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 oclachlorobiphenyls produce hepatocellular carcinomas in rodents should be explored. Particularly in the United States, mixtures such as Aroclor 1242, 1254, and 1016 were used more than Aroclor 1260. Because of these differences in potency, (he PCBs in heavily contaminated areas of our environment should be characterized according to their isomeric composition. For instance, whether the PCBs in Lake Michigan are 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). In some species they may cause atrophy of the thymus, intrahcpalic bile duct hyperplasia, hyperplasia of the epithelial lining of the urinary bladder, atrophy of the sebaceous glands, and hyperkeratosis of the ducts (40). Some isomers arc fctotoxic, 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 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 i 94 KIMBROUGH HUMAN HEALTH EFFECTS OF PCBs & PBBs 95 mechanism by which Ihese types of chemicals affect hepatic retinoids is not clear. Apparently, the duration of the reduction of hepatic retinoids does not correlate with the induced aryl hydrocarbon hydroxylase (Allll) activity (43). levels were lower in females of each age group. Similar findings were made for DDT residues. The scrum cholesterol level was positively associated with the log PCB level, independent of age, sex, fish consumption, body-mass Body Burdens index, and alcohol consumption. Rales of borderline and definite hyperten sion for study participants were 30% higher than those expected on the basis Many investigators have reported PCBs in human tissues (44 , 45). In the of national rates (54). Log PCB serum values contributed significantly to United States, according to data from the Centers for Disease Control (CDC), explaining the variability of log systolic and diastolic blood pressure in mean PCB serum levels are about 5-7 hg/ml (pbb), although some patients multiple regression analysis (55). Median total cholesterol levels of in may have higher serum levels without any documented unusual exposure. dividuals in the United States increase with age from about 150 to 160 mg/dl These data were also summarized by Kreiss (46). Levels in adipose tissue and al age 20 to over 200 mg/dl at age 50. PCBs in blood are influenced by serum in human milk fat are 100-200 times as high, since PCBs are highly lipid lipid content, and populations with inherently lower total serum cholesterol soluble (20). Mes et al (47) reported that PCB levels in adipose tissue of levels appear to have a different PCB scrum to adipose tissue ratio. The accident victims ranged from 0.9-9.4 mg/kg. age-associated increase in blood PCB levels could be related to the long Sahl el al (48) surveyed PCB blood levels in 738 pre-employed and 1,058 half-life of some PCB isomers that are preferentially retained in mammals currently employed workers of a utility company. The median blood level (29); as long as exposure continues, a true steady state between intake and before employment was 4 mg/I and the range, 1-37 mg/I. These levels were excretion is never reached. Other variables affecting body burdens may be quite similar to those in the currently employed group. differences in metabolism with age. In the Triana studies, the blood levels of Patients who died of cancer in Denmark had somewhat higher levels of total DDT residues also increased with age, and others have made similar PCBs in their adipose tissue (49). Since terminal cancer patients have usually observations (56. 57). Lawton et al (58) studied workers who had been lost a great deal of weight, bioconcentration may have occurred. Similarly, in exposed to electrical-grade Aroclor 1016, 1242, and/or 1254; the study patients with highly impaired liver function, tissue concentrations of xenobi- covered the period from before the workers were exposed to two years after otics may be slightly higher than those in healthy persons. However, levels PCB exposure ceased. Serum levels for the lower chlorinated PCBs in 1977 remained higher if parameters such as weight, height, occupation, and resi ranged from 57-2270 ppb and in 1979, from 12-392 ppb; for the higher dence were considered (50), whereas levels of PCBs in breast fat tissue from chlorinated PCBs scrum levels ranged from 6-142 ppb in 1977 and from patients with breast cancer were similar to those of controls (51). 4-108 ppb in 1979. These findings again illustrate the preferential excretion Furthermore. Lawton el al (52) demonstrated that random errors and of lower chlorinated PCB. Lawton el al (58) also found that cholesterol levels i interlaboralory variations in procedure and methods of data reporting can correlated with log serum PCBs. Similar associations with log scrum PCBs influence serum and adipose PCB levels. Unless an interlaboratory quality- were found for log gamma glutamyl transpeplidasc (GGTP) and, in some control system is set up, measured levels between laboratories are not neces cases, for log alanine aminotransferase. When (he PCB concentrations were sarily comparable. For instance, Lawton et al (52) found that the results of expressed as levels in serum lipids, all the associations between scrum lipids repeated analyses on serum samples of known composition showed the 95% or enzymes and log serum PCBs disappeared except for those between log prediction interval for an individual measurement to be about 42%. This GGTP and log PCBs. Similarly, Chase et al (59) found no significant interval depends on the method of extraction, the procedure used, and the correlation between either serum triglycerides or aminotransferases and the means of quantitation. PCB levels in adipose tissue. How age and length of exposure affect these Summary of Human Epidemiology Studies Recently, investigators studied the predominantly black population of Triana, i a small rural town in the southern United States (53). This population was excessively exposed to DDT residues by consuming contaminated fish. The o o o 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 parameters is not adequately explained in the article. Finally, Akagi St Okumura (60) were not able to 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 ex plained by the increased solubility of PCB in serum with higher lipid content. In several cross-sectional studies of exposed workers, only minor abnormalities not necessarily related to PCB exposure have been detected ID STLCOPCB4029233 96 KIMBROUGH (62-66). In cross-seclional 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-glutamyllranspcplidasc--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 Ihe chloracne cyst, which is skin colored and measures from 1-10 mm in diameter, with 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 Ihe 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 & Alden (68) examined 17 of 23 workers engaged in the production of PCBs. The workers had chloracne involving Ihe face, genitalia, trunk, and extremities. Before Ihe outbreak of chloracne in Ihe plant, the electrical property 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 heated (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 el 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/m3. Evidence of slight liver injury was also present. Ouw ct 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 pph (Mg/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 dichlorobiphcnyls. Irichlorobiphenyls, tetrachlorobiphenyls, and penlachlorobiphcnyls--plasma antipyrinc half-life was significantly lower than that in matched controls, suggest ing the induction of mixed-function oxidases in the liver. IJiesc workers had been exposed to Aroclor 1016 for at least two years and had no obvious symptoms of PCB poisoning. Other health effects arc eye and upper respiratory irritation. Warshaw et al (65) 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. Spirometric studies were conducted on 309 workers; 66 of them were dropped from the study because they 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, Ihe 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 are difficult to interpret. Taylor et al (73), in an attempt to determine whether the fetus would be affected in capacitor workers, examined pregnancy outcome and birth weight, and found that 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 association 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 & Gladcn (77), for instance, found that mothers with high levels of DDE |l,r-(2.2-dichloroethenylidene)-bis-4-chlorobenzene| in their milk tended to wean their 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 clear. In animals, DDT 98 KIMBROUGH homologs, but nol specifically DDK, have been shown lo have cslrogenic 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 et al (80) reported results of a preliminary study of a group of 51 research and development employees and 41 refinery plant employees al a New Jersey petrochemical facility. Between 1949 and 1957 these workers had been exposed lo 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 combined. 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 (SMK) 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 reported on the death certificates. However, a review of the medical records raises questions about at least one of these tumors. Bertazzi et 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 significant!y 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 qualerphenyls (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 repeated 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 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 1*08 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 lo be 2.000-3,000 ppm. Kanechlor 400 had been used for heating the rice oil in a metal container at over 200C, al a reduced pressure of 3-44 mm llg, lo 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 reanalysis 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 lo Buser et al (86). the Yusho oil contained more than 40 polychlorinated dibenzofuran isomers, including the highly toxic 2,3,7,8-tctrachlorodibenzofuran (TCDF) and 2.3,4,7.8-pcntachlorodibenzofuran (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 fig/kg body weight/d. PCB, 0.9 ^ig/kg body weight/d. PCDF, and 148 p.g/kg body weighed. 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 conjunctivac, jaun dice. swelling of the upper eyelids, a feeling of weakness, numbness of the limbs, and fever. Over I,(XX) people were affected. Thirty-six babies showed fetal PCB syndrome, which consists primarily of a dark-brown pigmentation 100 KIMUROUGli of the 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 time the babies were between two and five months old. In affected infants, the face wax edematous, and spotty calcifications were noticed in the parietal and occipitul arcus 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 persisted, and the patients have chronically infected airways. In the early 1970s, some changes were noted in the patients' serum immunoglobulin levels, but the levels returned to normal. Over lime the severity and the extent of the skin lesions improved considerably in the exposed population. Fifteen years after the accident, only a very few patients had extensive chloracne (91). About five years after the outbreak of Yusho, tissue and body fluids of Yusho patients were analyzed for various congeners of PCB and PCDF. At this lime, the PCB levels in adipose tissue were 1.9 1.4 ppm (mg/kg). In the liver they were 0.08 0 06 ppm and in blood, 6.7 S.3 ppb (/^g/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 the 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 the other iso mer* was retained in the patients' tissues. In Ihe five patients studied, the concentration of this isomer ranged from 6.9 ppb (/tg/kg) in a specimen obtained in 1969 to 0.1 ppb (Mg/kg) in a specimen collected in 1977. Measurable concentrations of TCDFs were only detected in the earlier years. Although not the most toxic isomer, Ihe 2.3.4,7,8-PCDF caused mixedfunction oxidase induction at a dose of I pg/kg in rats, and atrophy of the thymus, suggesting toxicity at a very low dosage level. Titus, the clinical oi manifestations observed in these patients were primarily caused by the PCDFs, specifically by the more toxic isomers. o In the Yucheng episode, it was never determined with certainty how the o rice oil was contaminated (93). In 1979 a school for blind persons informed a */J local health bureau in Taichung County that a strange disease characterized by an acnclike skin eruption had been occurring frequently among students and HUMAN HEALTH EFFECTS OF PCBs & PBBs 101 staff since the end of March. At the same time. 85 of 150 workers in a nesrby plastic shoe factory had the 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 the outbreak. 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 the disease resembled the Yusho disease in Japan, sample: of C-rice oil and patients' blood were analyzed in Japan and were found It contain either a Kanechlor-400 or a Kanechlor-500 mixture at concentration: as high as 65 and 108 ppm (mg/kg), respectively. Over 2.000 patients wen finally identified as having been poisoned by contaminated rice oil. Oi samples collected from other outbreaks contained PCBs at concentrations o 31-300 ppm (mg/kg). Retrospective studies determined that the period o PCB intake ranged from 3 to 9 months. The average total intake for eacl person varied from 0.77 to 1.8 mg of PCB. Within the first year of th outbreak, the blood levels of PCB in 13 patients ranged from 3 ppb to 1,15' ppb. Most of the patients had blood levels between 11 and 150 ppb (pg/kg) Ihe symptoms observed in these patients were quite similar to those alread described for the patients in Ihe 1968 Yusho outbreak in Japan. The rice oil was not only contaminated with PCBs but also with PCDFs an: polychlorinated quaterphcnyls. It contained the same major components o PCDFs observed in the rice oil in Japan--namely. 2,3.4.6,7-PCDF an 2.3.4.7.8- PCDF. Relatively high concentrations of 2,3.4.5.3',4'-hexachlo robiphenyl were found in Ihe blood and adipose tissue of the Yuchen patients. This particular PCB isomer is biologically quite active, and th concentration of 2.3,4.3'4,-penlachlorobiphenyl was also elevated in thes patients. Furthermore, as in the Yusho patients, the concentration of 2.3,7,8 TCDF was comparatively low (92). Chen et al (94) analyzed addition! samples of the oil, blood, and adipose tissue of the Yucheng patients. Thes investigators identified several TCDF and PCDF isomen. Apparentl) 2.3.7.8- TCDF was only a minor component in Ihe oil; the major compon was 2,3,4,8-TCDF. One of (he major furans in the toxic oil was 2,3,4,7.f PCDF. Ihe concentrations of the PCDFs in different oil samples ranged from 0.2 to 1.68 ppm. Polychlorinated quaterphcnyls were present in concentratior ranging from 25 to 53 ppm (mg/kg). Overall. Ihe concentrations of PCD! and PCQs were lower in these oil samples than in the oil samples that h caused the Yusho outbreak. Whether these oil samples were representative not really known. The 3.4,3'4'-tetrachlorobiphenyl was also identified in tf oil that caused Yucheng disease in Taiwan. This isomer is considered to t i O 102 KIMBROUGH the most toxic PCB isomer present in commercial PCB preparations (95); it was present at a concentration of about 1%. Mosl other commercial PCB preparations, such as the Aroclors. in the United Slates have not been shown to contain (his 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 the 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 dental 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 et at (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 streptodomase subcutaneously into the flexor side of the forearm. The response was read at 24 hours (hr) and again at 48 hr after injection. Eighty percent of the 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 the 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 the 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 the 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 the 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. (It 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 et 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/m1 air. PCDFs were also formed. The soot samples contained letrachlorodibenzofuran up to 90 /xg/g, of which 6.5 /xg/g was 2,3,7,8-tctrachlorodibcnzofuran. In addition, monochloropyrencs and dichloropyrcnes were found. Most of the men had a transient sensory neuropathy in their lower extremities. Chang et al (104) reported increased urinary 6-aminolcvulinic 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 slate of Michigan. Most persons living within the lower peninsula of Michi- ly O t'0 0 0 104 Kimbrough gan have had slight exposure, since the contamination resulted from dairy products and since normal marketing channels for these prtRlucts 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 serum 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 ppb (pg/kg); in serum it was 1.3 ppb (/zg/kg) for adults and 1.8 ppb (/c/gkg) for children. The highest adipose tissue concentration was 37 ppm (mg/kg) (18). In addi tional studies, when cohorts of PBB-exposed residents of Michigan were compared with residents of the slate of Wisconsin, a higher prevalence of a variety of symptoms and complaints was noted in the Michigan residents (106). Similarly, in comparative neurobehavioral 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 scrum PBB levels in this entire group ranged from no detectable levels to 1,900 ppb (ng/l), with a mean of 21.2 ppb tng/l) and a median of 3 ppb (/zg/l). 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 scrum than in those with measurable quantities. In atl groups, including chemical workers and quarantined farm j residents, the highest prevalence rates were in persons with the lowest scrum 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 ^nd 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 scrum, which indicated partial placental passage. Human milk contained PBBs at 107-119 times the quantity found in maternal serum. PBBs were also detected 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 had 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 that PBBs are very slowly excreted, which is consistent with the findings of Tuey & Matthews in rats (110). 7'he 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 tests were conducted, blood pressure was measured, and height and weight were determined. In this study, 83 participants had PBB serum levels of 50 ppb (^ig/l) or more. In the middle group. 83 had PBB levels of 5-49 ppb and 96 had PBB levels of 0-44 ppb (/xg/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 the low PBB group, too few were using them to affect urine porphyrin levels.) In none of a variety of other studies conducted on this population as well as other groups in Michigan did any findings indicate that exposure In PBB had impaired the health of the exposed group. All of these studies have been reviewed by Fries (105). STLCOPCB4029239 Q o o 106 KIMBROUGH Although this population was exposed during a 9-month period in 1973 and 1974, whether it will have chronic health effects is unknown. This particular cohort needs to be followed for 30 to 40 years before the question of chronic health effects can be intelligently addressed. Two problems with assessing chronic health effects are that the cohort, in spite of its size, is still relatively small and that the amount of exposure it has received varied widely. Although some members of the group exposed to 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 that 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 the mean was 170 mg per person. In rats given 200 mg/kg, a dose that 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 to the toxic effects of PBBs and whether their response would be similar lo that of rats is not known. In conclusion, various toxic effects of PBBs and PCBs have been described in laboratory animals. In humans, acute poisoning outbreaks have only occurred following exposure to a combination of PCBs and PCDI-'s. When humans were exposed only to PCBs or PBBs, the only observed acute effects have generally been minor. So far, no significant chronic health effects have been causally associated with exposure to PCBs or PBBs. Use of trade names is for identification only and does not constitute endorsement by the Public Health Service or the US Department of Health and Human Services. Utrralure Cited 1. Jensen. S. 1966 Report of a new chemi cal hazard. New Sci. 32:612 2. 1982. Polychlorinated biphenyl* (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in electrical equipment. Fed. Begin 47.37342-1*) 3. 1982. Polychlorinated biphenyls (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in closed and controlled waste man ufacturing processes. Fed. Regut. 47:46980-96 4 McLeod. K E 1981 Polychlorinated biphenyl* in indoor air. Environ. Sci. Technot. 15.926-28 5. Kut/. F. W . Yang. H S C 1976 A note on polychlorinated biphenyl* in air. In Proc. Nutt. Con/. Polvchlttrinnietl Btphenvls, Chicago, 197$, EPA-560/67>-<XH, p. 182. Washington. |)C: En viron. Prof. Agency 6. Cnrdlc, F.. Locke. R-. Springer. J. 1982- Determination of human risk in regulating polychlorinated htpltenyl* (K'lld-i ca*c Mudy. CRC Toxicol. Rnk /turn, 2.211 25 7. Cnrdlc. F.. Locke. R.. Springer. J. 1982 Risk astessment in a federal reg ulatory agency: an a*&csunent of ri*k associated with the human consumption of some specie* of fish contaminated with polychlorinated biphenyls (PCBs). Environ Ifcntih Prrspecl. 4.V171-H2 HUMAN HEALTH EFFECTS OF PCBs A PBBs 107 8. Sullivan. J. R., Delftno. J. J., Huelow. C. R . Sheffy. T. B. 1983. Polychlori nated biphenyls in the fish and sediment of the Lower Fox River. Wisconsin. Bull. Environ. Contain. Toxicol. 30:38 64 9. Schwartz. P. M.. Jacobson. S. W.. Fein, G. G.. Jacobson. J. L., Price. H. A. 1983. Lake Michigan fish consump tion as a source of polychlorinated biphenyls in human cord serum, mater nal scrum, and milk. Am. J. Public Health 73:293-96 10. Taisukawa. R, 1976, PCB pollution of the Japanese environment. In PCB Poisoning and Pollution. ed. K. Higuchi. pp. 147-79 Tokyo: Kodansha 11. Morita. M., Nakagawa. J.. Rappe. C. I97K. Polychlorinated dibenzofuran (PCDF) formation from PCB mixture by heat and oxygen. Bull. Environ. Contom. Toxicol. J 9:665-70 12. O'Keefe. P. W . Silkworth. I. B.. Gierthy. J. F., Smith. R. M.. DcCaprio. A. P . ct al. 1985. Chemical and biological investigations of a transformer accident at Binghamton, New York. Environ. Health Perspect. 60:201-9 13. Hul/inger. O., Ghulam, G. C.. Brock. <. C.. Johnston. 1.. E. 1985. Formation of polychlorinated dibenzofuran* and di oxins during combustion, electrical equipment fires and PCB incineration. Environ. Health Perspect. 60:3-9 14. Carter. L. 1976. Michigan PBB in cident. Chemical mix-up leads to disas ter. Science 192:240-43 13. Willett. L. B.. Liu. T. T. Y., Durst. H. I.. Cardwell. B. D., Renkie. E. D. 1985. Quantification and distribution of polychlorinated biphenyls in farm silos. Hull. Environ. Contam. Toxicol. 33:31 60 16. Kimbrough. R. D. 1983. Laboratory and human studies on polychlorinated biphenyls (PCBs) and related com pounds. Environ. Health Perspect. 59:99-106 17. Smith. A. B.. Schloemer. J.. Lowry, L. K , Smallwood. A. W., Llgo, R. N.. ct al. 1982. Metabolic and health con sequences of occupational exposure to polychlorinated biphenyls (PCBs). Br. J fnd. Med. 39:361-69 18. Wolff, M. S . Anderson, H. A.. Sellkolf. J. J. 1982. Human tissue burdens of halogenated aromatic chemicals in Michigan. J. Am. Med. Assoc. 247: 2112-16 19. Eysler, J. T.. Humphrey. H. E. B.. Kimbrough, R. D. 1983. Partitioning of polybrominated biphenyls (PBB) in serum, adipose tissue, breast milk, placenta, cord blood, biliary fluid and feces. Arch Environ. Health 38:47-33 20. Jensen. A. A. 1983. Chemical contami nant* in human milk. Residue Rev. 89:1-128 2L Jensen. R. G.. Clark. R. M.. Ferris, A. M. 1980. Composition of the lipids in human milk, a review. Lipids 15:345-55 22. Landrigan. P. J., Wilcox, K. R. Jr., Silva, J. Jr.. Humphrey, H. E. B., Kauffman. C.. Heath. C. W. Jr. 1979. Cohort study of Michigan residents ex posed lo polybrominated biphenyls: epidemiologic and immunologic find ings. Ann. N. Y. Acad. Sci. 320:284 94 23. Kuratsune. M.. Yoshimura. T.. Mattuzaka. J.. Yamaguchi. A. 1972. Epide miologic study on Yusho, a poisoning caused by ingestion of rice oil con taminated with a commercial brand of polychlorinated biphenyls. Environ. Health Perspect. Exp. No. 1:119-28 24. Yoshimura. T.. Hayabuchi, H. 1985. Relationship between amount of rke oil ingested by patients with Yusho and their subjective symptoms. Environ. Health Perspect. 59:47-51 25. Kimbrough. R. D . Buckley. J . Fishbcm. L., Flamm. G.. Kasza. L., et al. 1978. Animal toxicology. Environ. Health Perspect. 24:17.3-85 26. Vos. J. G.. Faith. R. E.. Luster. M. I. 1980. Immune alterations. In Halogen ated Biphenyls, Terphenyls, Naph thalenes. Dlbenxodioxins and Related Compounds. Topics in Environmental Health, ed. R. D. Kimbrough, 4:241-58 Amsterdam: Elsevier Biomedical Bio. 27. Linder. R. E.. Gaines. T. B., Kim brough. R. D. 1974. The effect of polychlorinated biphenyls on rat re production. Food Cosmet, Toxicol. 12: 63-77 28. McNulty. W. P., Becker. O. M.. Cory. H. T. 1980. Chronic toxicity of 3.4.3'4*- and 2,5.2,.5'-telrchloroblphenyls in rhesus macaques. Toxicol. Appl. Pharmacol. 56<2): 182-90 29. Matthews. H. B., Anderson, M. W. 1975. Effect of chtorinslion on the dis tribution and excretion of polychlori nated biphenyls. Drug. Metob. Dispos. 3:371-80 30. Matthews. H. B.. Kalo. S. 1979. The metabolism and disposition of halogen- ated aromatics. Ann. N. Y. Acad. Sci. 320:131-38 31. Parkinson. A., Robertson. L.. Uhlig. L.. Campbell. M A.. Safe. S. 1982. 2.3.4.4,,5-Peniachlorobpbcnyi: differ- XOVOOO 108 KIMBROUGH cnlial effect, on C57DIJ6I <nd IMIAJ2J phenyl. Toxicol. Appl. Pharmacol. 78: inbred mice. Bioxhem. Pharmacol. 180 89 31:2830 33 44. Kills. I:. W.. Slrassman. S C. 1976. 32. Poland. A.. Clover, fi. 197?. Chlori- Residues of polychlonnated biphenyls in naied biphenyl induction of trylhydro- the general population of the United carbon hydroxylase activity: a study of States. Sec Kef. 5. pp. 139-48 structure activity relationship. Mol. 45. Juskiewic*. T., Niewiadowska. A.. Pharmacol. 13:924-38 Radomanski, T. 1977. Polychlorinated 33. Schaeffer. E.. Greim. II., Goessncr. W. biphenyl residues in human adipose tis 1984. Pathology of chronic polychlori sue. Pol. Ty*. l.ek. 32:173-175 nated biphenyl (PCR) feeding in rats. 46. Krciss. K i9H5. Studies on populations Toxicol. Appi Pharmacol. 75:278 288 exposed to polychlonnated biphenyls. 34. Nurback. 1) H , Wellman. R. II 1985. Environ. Health Perspect. 60:193-199 Polychlorinated biphenyl induction of 47. Mes. J . Davies. D.J .Turton.D 1982. hepatocellular carcinoma in the Sprague- Polychlorinated biphenyl and other Dawlcy rat. Environ. Health Perspect. chlorinated hydrocarbon residues in adi 60:97-105 pose tissue of Canadtans. Bull. Environ. 35. Kimbrough. R. D.. Squire, R. A.. Lin Contam. Toxicol! 28:97-104 der. R. E . Strindberg. J. D . Montali, 48. Sahl. I. D., Crocker. T.. Gordon, R. L. R. J . Burse. V. W. 1975. Induction of Facdcr. I:. J. 1985 Polychlorinated fiver tumors in Sherman strain rats by biphenyl* in the blood of personnel ftom polychlonnated biphenyl Aroclor 1260. an electric utility. J. Occup. Med. ), Natl. Cancer Inst. 55:1453-59 27:639-43 36. National Cancer Institute. 1977. Bioas 49. Unger. M., Olsen. ). 19R0. Organochlo- say of Aroclor (frademart) 1254 for line compounds in the adipose tissue of possible carcinogenicity. Springfield. deceased people with and without can VA: Natl. Tech Inf. Scrv. as PB 279 cer. Environ. Res. 23:257-63 624/IGA. NCI-CO TR-38. Chem. 50. Unger. M . Olsen. J . Clausen. 1 1982. Abstr. Serv. No. 27323 18 8. DHEW Organochlorinc compounds in the adi Publ. Nil! 78 838 pose tissue of deceased persons with and 37. Ward. J. M. 1985. Proliferative lesions without cancer: a statistical survey of of the glandular stomach and liver in F some potential confourulers. Environ. 344 rats fed diets containing Amclor Res. 29:371-76 1254. Environ. Health Perspect. 60:89 51. Unger. M . Kiaer. II.. Blichen-Tnfi, 96 M . Olsen. J.. Clausen. J. I9H4. Or- 38. Morgan, R. W . Ward. J. M . Man- ganochtorinc compounds In human mann, P. E. 1981. Aroclor 1254 Induced breast fat from deceased with and with Intestional metaplasia and adenocarcino out breast cancer and in a biopsy materi ma in the glandular stomach of F344 al from newly diagnosed patients un rats. Cancer Res. 41:5052-59 dergoing breast surgery. Environ. Res. 39. Safa. S., Bandkra. 5., Sawyer. T., 34:24-28 Robertson. L., Safe, L., ct at. 1985. 52. Lawton. R. W.f Brown. J. F.. Ross. M. PCB: structure-function relationships R., Feinaold. I 1985- Comparability and mechanism of action, Environ, and precision of scrum PCB measure Health Perspect. 60.47-5h ments. Arch. Environ. Heahh 40:29 40. McConnell. C. E. 1980. Sec Ref. 26. 37 pp. 109-90 53. Kreiss. K., Zack, M.. Kimbrough. R. 41. McNulty. W. P. 1983. Tosicity and I)., Needham, L. L., Smrek. A. L.. feto<os idty of TCDD. TCDF. and PCB loneI. B. T. 1981. Cross-sectional study isomers in rhesus macauuea (Mocaca of a community with exceptional expo mulottal. Environ. Health Perspect. sure to DDT. J. Am. Med. Assoc. 60:77-88 245:1926-30 42. Thunberg. T. 19*4. Effect of TCDD on 54. National Center for Health Statistics. I vitamin A and iu relation to TCDD toaidly. Banbury Rep. IB: Biol. Mrch. 1978. Blood pressure levels of persons 6-74 years. US. 1971-1974. HRS 78 Dioxin Action. New Yost: Cold Spring 1648. scries tl. No. 203. Hyattsvtlle, Harbor Lab Md, Nall. Cent. Health Stat.. DHEW 43. Brouwer. A.. Van den Berg. K. J.. Kuk- 55. Kreiss. K.. Zack. M., Kimbrough. R. kr. A. 1985. Time and dose responses I).. Needham, L. L., Smrek. A L., of the reduction in retinoid concen Jones, B. T. 1981. Association of blood trations in C57BI/Rij and DBA/2 mice pressure and polychlorinated biphenyl induced by 3,4,3'4,-letrachlon>bi levels. J. Am. Med Assoc. 245.2505-9 HUMAN HEALTH EFFECTS OF PCBs A PBBs 109 56. Kut?. F. W.. Yobs. A. R.. Slrassman. S. C.. Viar. I. F. 1977. Effcctsof reduc ing DDT usage on total DDT storage in humans, Pestic. Monit. J 11:61-63 57. Davies. I. K.. Edimmdson. W. F.. Raf* fonclli. A., et at. 1972. The role of so cial class in human pesticide pollution, i Am. J. Epidemiol. 96:334-41 58. Lawton. R. W , Ross. M. R . Feingold. J . Brown. J. F. 1985. Effects of PCB exposure on biochemical and hemato logical findings in capacitor workers Environ. Health Perspect. 60:165-84 59. Chase. K. H., Wong. O.. Thomas, D., Stai. B. W.. Bcmey. B. W.. Simon. R. K 1982. Clinical and metabolic ithnormalilics associated with occupa tional exposure to polychlorinated bi phenyls. J. Occup. Med. 24:109-14 60. Akagi. K.. Okumora. M. 1985. Association of blood pressure and PCB level in Yusho patients. Environ. Health Perspect. 59:37-39 61. Brown. J. F. Jr. I9K4. Polychlorinated btphcnyl /1`CB) partitioning between adipose tissue and strum. Bull. Environ. Crmtam. Toxicol. .3.3:277-80 62. Emmett. I;. A. 1985. Polychlorinated biphenyl exposure and effects in trans former repair workers. Environ. Health Perspect. 60:185-92 63. Humphrey, H. E. B.. Price. H. A., fludd. M, f. 1976. Evaluation of changes of the level of polychlorinated biphenyls (PCBI in human tissue. Hnal Rep. EDA Contract No. 22J-7J-2209. Washington. DC: DHEW, FDA 64. Baker, E. L. Jr.. Landrigan. P. J.. Ghicek. C. J . Zack. M. M Jr . Litldle. J. A., at al 1980 Metabolic con sequences of exposure to polychlori nated biphenyls in sewage sludge. Am. J. Epidemiol. 112:553-63 65. Wurshaw. R., Fiihhcln. A.. Thomlim. J.. Miller. A.. Sclikoff. I. J. 1979. De crease in vital capacity in PCB exposed workers in a capacitor manufacturing facility. Ann. N. Y. Acad. Scl. 320:277 8.3 66. Sak. M., Ahlcrs. I. 1977. Serum lipid changes under conditions of occupation al exposure to chlorinated biphenyl*. OrA. Dermatol. 52:62-65 67. Kimbrough. R. 0. 1980. Occupational exposure. See Ref. 26, pp. .373-97 68. Jones. J. W.. Alden. H. S. 1936. An acncform dermatergosis. Arch. Drrmatol. Sxphdof. 33:1022-34 69. National Institute for Occupational Safety and Health (NIOSII). 1977. Criteria for a recommended standard. Occupational exposure to polychlori nated biphenyls (PCBs). Washington, IX?: Superintendent of Documents. US Govt. Printing Office. USDHEW (NIOSII) Publ. No. 77-225 70 Mc.gs. J. W.. Album. J. I.. Karlin. B I 1954. CTiktracne from an unusual ex posure to Arochlor. J. Am. Med. Assoc. 154:1417-18 71. Ouw, H K., Simpson. G. R., Siyalt. D. S. 1976. Use and health effects of Aro- clor 1242, a polychlonnated biphenyl in an electrical industry. Arch. Environ. Health 31:189-94 72. Alvares. A. P., Fischbcin, A.. An- derNon. K. E.. Kappas. A. 1977. Altera tions in drug metabolism in workers ex posed to polychlorinated biphenyls. Clin. Pharmacol. Ther. 22:140-46 73 Taylor, P. R.. Lawrence. C. L., Hwang. H. I... Patterson, A. S. 1984. Polychlo rinated biphenyls influence on birth- weight and gestation. Am. J. Public Health 74:1153-54 74. Jacobson. J. L.. Jacobson. S. W.. Schwartz, P. M. Fein. G. G., Dowler, J K. 1984. Prenatal exposure to an en vironmental toxin. a test of the multiple effects model. Dev. Psychol. 20:523-32 75. Fein, G. G., Jacobson. J. L., Jacobson. S. W . Schwartz. P. M.. Dowler. J. K. 1984. Prenatal exposure to polychlori nated biphenyls: effects on birth sice and gestational age. J. Pediatr. 102:313-20 76. Jacobson. S. W., Jacobson. J. L., Schwarts. P. M.. Fein. G. G. 1983. Intrauterine exposure of human new borns to PCBs: measures of exposure. In PCBs: Human and Environmental Hazards, cd. F. M. D'ltri, M. Ksmrin, pp 311-43. Boston: Butterworth 77. Rogan. W , Oladen. fi. 1982. Duration of breast-feeding and environmental contaminants in milk. Am. J. Epidemitd. II6 565A 78. Kogan. W. J.. Gladen. B. C.. McKin ney. J. D., Carreras, N.. Hardy. P.. et al. I9K6. Polychkxinated biphenyls (PCBs) and dichlorodipbcnyl di* chlomcthcne (DDE! in human milk: effects of maternal factors and previous lactation. Am. J. Public Health 76:172 77 79. Gcllcrt, R. J., Heinrichs. W. L.. Swerdloff. R. S. 1972. DDT homologs. atroten-like effects on the vagina, uterus, and pituitary of the rat. Endocrinology 91:1095-100 *0. Hahn, A. K.. Rosenwaike, I.. Herr' mann. N., Grover. P., Stellman. J.. O'Leary, K. 1976. Melanoma after ex posure to PCB. N. En/tl. J. Med. 295:450