Document aBR10jvxGeM6YRV9QrDgbEb2y

Sponsor: APME Ad-Hoc APFO Toxicology Working Group FINAL REPORT Study Title: 4-Week Capsule Toxicity Study with Ammonium Perfluorooctanoate (APFO) in Cynomolgus Monkeys Author: Peter J. Thomford, PhD Study Completion Date: December 18, 2001 Performing Laboratory: Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 Laboratory Project Identification: Covance 6329-230 Sponsor Project Identification: 3M Study No. T-6889.2 Page 1 of 159 Covance 6329-230 3M T-6889.2 COMPLIANCE STATEMENT 4-Week Capsule Toxicity Study with Ammonium Perfluorooctanoate (APFO) in Cynomolgus Monkeys All aspects of this study were in accordance with the Environmental Protection Agency Good Laboratory Practice Standards (GLPs), 40 CFR 792, except that bile acid determinations done by the University of Dundee were not done in compliance with GLPs. 2 Covance 6329-230 3M T-6889.2 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Covance Laboratories Inc., in accordance with the Environmental Protection Agency (EPA) Good Laboratory Practice Standards, 40 CFR 792. The following inspections were conducted and findings reported to the study director and study director management. Inspection Dates From To 06/07/98 06/07/98 07/16/98 07/16/98 09/22/98 10/01/98 10/01/98 10/01/98 09/15/98 10/02/98 06/10/99 06/10/99 12/07/99 12/07/99 02/23/01 02/23/01 11/27/01 11/27/01 Phase Protocol Review Clinical Laboratory Inspection Report Review Protocol Amendment Review Data Review Protocol Amendment Review Protocol Amendment Review Report Review Report Review Date Reported to Study Director and Study Director Management 06/08/98 07/16/98 10/01/98 10/01/98 10/02/98 06/10/99 12/07/99 02/23/01 11/28/01 3 Covance 6329-230 3M T-6889.2 STUDY IDENTIFICATION 4-Week Capsule Toxicity Study with Ammonium Perfluorooctanoate (APFO) in Cynomolgus Monkeys Test Material Ammonium Perfluorooctanoate (APFO) Sponsor APME Ad-Hoc APFO Toxicology Working Group Study Monitor Paul Lieder, PhD, DABT 3M Toxicology Services Building 220-2E-02, 3M Center St. Paul, Minnesota 55144-1000 612.737.2678 Alternate Study Monitor John Butenhoff, PhD, DABT 3M Toxicology Services 612.733.1962 Study Location Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 Study Director Peter J. Thomford, PhD Covance Laboratories Inc. P.O. Box 7545 Madison, Wisconsin 53707-7545 608.241.7207 Study Timetable Study Initiation Date In-Life (Experimental) Start Date In-Life Termination Date Experimental Termination Date June 11, 1998 June 17, 1998 July 16, 1998 December 18, 2001 4 KEY PERSONNEL Covance 6329-230 3M T-6889.2 Study Director Study Toxicologist Study Coordinator Supervisor, Large Animal Toxicology Supervisor, Dose Formulation Supervisor, Laboratory Animal Medicine Clinical Pathologist Supervisor, Clinical Pathology Anatomical Pathologist Supervisor, Anatomical Pathology Peter J. Thomford, PhD Dale Aldridge, BS Patricia K. McKee Pesik, BS, LAT Meechelle Bordeaux, LAT Dixie Bushee, BS, LATG Donna J. Clemons, DVM, MS Diplomate, ACLAM Robert L. Hall, DVM, PhD Diplomate, ACVP (Clinical Pathology) Ronald Markevitch, BS, MT (ASCP) James L. Carter, DVM, PhD Diplomate, ACVP Laurie J. Schuller, BA, LAT 5 Covance 6329-230 3M T-6889.2 CONTENTS Page ABSTRACT.................................................................................................................... 9 PURPOSE..................................................................................................................... 11 REGULATORY COMPLIANCE.................................................................................. 11 TEST MATERIAL........................................................................................................ 11 Test Material ............................................................................................................. 11 Reserve (Archive) Samples........................................................................................ 11 Disposition ................................................................................................................ 11 TEST SYSTEM ............................................................................................................ 12 Test Animal............................................................................................................... 12 Identification ............................................................................................................. 12 Acclimation............................................................................................................... 12 Housing and Maintenance ......................................................................................... 12 Justification ............................................................................................................... 13 PROCEDURES............................................................................................................. 13 Group Designations and Dose Levels ........................................................................ 13 Dose Preparation ....................................................................................................... 14 Dose Analyses........................................................................................................... 14 Method of Administration ......................................................................................... 14 Clinical Observations ................................................................................................ 15 Body Weights............................................................................................................ 15 Blood Hormone Determination.................................................................................. 15 Serum APFO Level Determination ............................................................................ 16 Clinical Pathology..................................................................................................... 16 Additional Blood Collection ...................................................................................... 17 Necropsy ................................................................................................................... 17 Palmitoyl CoA Oxidase Determinations .................................................................... 17 Cell Proliferation Evaluation ..................................................................................... 18 Receptor Level Determination ................................................................................... 18 Bile Acid Determination............................................................................................ 18 Liver APFO Determination........................................................................................ 18 Tissue Preservation.................................................................................................... 19 Histopathology .......................................................................................................... 19 Statistical Analyses.................................................................................................... 19 RECORD RETENTION ............................................................................................... 20 6 Covance 6329-230 3M T-6889.2 CONTENTS (Continued) Page RESULTS ..................................................................................................................... 20 Clinical Observations ................................................................................................ 20 Body Weights............................................................................................................ 21 Food Consumption .................................................................................................... 21 Blood Hormone Analyses.......................................................................................... 21 Clinical Pathology ..................................................................................................... 21 Cell Proliferation Evaluation ..................................................................................... 22 Anatomical Pathology ............................................................................................... 22 CONCLUSIONS........................................................................................................... 22 SIGNATURES.............................................................................................................. 23 REFERENCES.............................................................................................................. 24 PATHOLOGY REPORT............................................................................................... 25 COMMENTS ON THE DATA ..................................................................................... 28 CODES, ABBREVIATIONS, AND UNITS.................................................................. 29 General Codes and Abbreviations.............................................................................. 30 Codes for Clinical Pathology..................................................................................... 31 Abbreviations and Units for Clinical Hematology...................................................... 33 Abbreviations and Units for Clinical Chemistry......................................................... 35 Codes for Anatomical Pathology ............................................................................... 37 TABLES 1 Summary of Clinical Observations - A.M./Weekly .............................................. 38 2 Summary of Clinical Observations - 30 Minutes Postdose ................................... 39 3 Summary of Clinical Observations - 60 Minutes Postdose ................................... 40 4 Summary of Clinical Observations - 90 Minutes Postdose ................................... 41 5 Summary of Body Weight Data (kg).................................................................... 42 6 Summary of Clinical Hematology Data - Day -8.................................................. 43 7 Summary of Clinical Hematology Data - Day -5.................................................. 45 8 Summary of Clinical Hematology Data - Day 30 ................................................. 47 9 Summary of Clinical Chemistry Data - Day -8..................................................... 49 10 Summary of Clinical Chemistry Data - Day -5..................................................... 52 11 Summary of Clinical Chemistry Data - Day 2 ...................................................... 55 12 Summary of Clinical Chemistry Data - Day 30 .................................................... 58 13 Incidence of Macroscopic Observations............................................................... 61 14 Incidence of Microscopic Observations................................................................ 62 7 Covance 6329-230 3M T-6889.2 CONTENTS (Continued) Page APPENDIX 1................................................................................................................ 63 Protocol..................................................................................................................... 64 Protocol Amendment No. 1 ....................................................................................... 81 Protocol Amendment No. 2 ....................................................................................... 86 Protocol Amendment No. 3 ....................................................................................... 88 Material Safety Data Sheet ........................................................................................ 91 Certificate of Analysis ............................................................................................... 98 APPENDIX 2.............................................................................................................. 100 Individual Animal Fate Data.................................................................................... 101 Individual Clinical Observations.............................................................................. 102 APPENDIX 3.............................................................................................................. 107 Individual Body Weight Data (kg)........................................................................... 108 APPENDIX 4.............................................................................................................. 109 Individual Clinical Hematology Data....................................................................... 110 Individual Clinical Chemistry Data.......................................................................... 119 APPENDIX 5.............................................................................................................. 131 Individual Animal Pathology Data........................................................................... 132 APPENDIX 6.............................................................................................................. 140 Quality Assurance Statement ................................................................................... 141 Summary and Individual Hormone Analyses Data................................................... 142 APPENDIX 7.............................................................................................................. 144 Cell Proliferation Report ......................................................................................... 145 8 ABSTRACT Covance 6329-230 3M T-6889.2 The purpose of this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study. In addition, the effect on critical enzyme levels, hormones, and other selected biochemical parameters was determined. Male cynomolgus monkeys were assigned to three groups (two animals in Group 1, three animals each in Groups 2 and 3). Animals in Groups 2 and 3 received gelatin capsules containing 2 and 20 mg Ammonium Perfluorooctanoate (APFO)/kg of body weight/day (mg/kg/day), respectively. Animals in Group 1 received empty gelatin capsules. Food was provided once or twice daily. Water was provided ad libitum. The animals were observed twice daily (a.m. and p.m.) for mortality and moribundity. At least once daily, animals were examined for abnormalities and signs of toxicity, and food consumption was assessed qualitatively. In addition, animals were observed approximately 30, 60, and 90 minutes postdose for signs of poor health or abnormal behavior. Body weight data were collected before initiation of treatment, on the first day of treatment, and weekly thereafter. Blood samples for hormone analyses [estradiol, estrone, estriol, thyroid stimulating hormone, total and free triiodothyronine (T3), and total and free thyroxin (T4)] were collected before initiation of treatment and on Day 30. Serum was harvested, and the samples were sent to Ani Lytics Inc., for analyses. Blood samples for cholecystokinin analyses were collected before initiation of treatment and on Day 30. Plasma was harvested, and the samples were sent to the DuPont Haskell Company for possible analyses. Blood samples for serum APFO concentration analyses were collected before initiation of treatment and on Day 30. Serum was harvested, and samples were sent to 3M for analyses. Blood samples were collected for hematology and clinical chemistry tests twice before initiation of treatment and on Day 30. In addition, blood for clinical chemistry tests was collected on Day 2. Additional blood was collected at the time of exsanguination, and samples were shipped to 3M for possible future analysis. On Day 30, the animals were anesthetized, weighed, exsanguinated, and necropsied. At necropsy, macroscopic observations were recorded, and selected tissues were collected and preserved. In addition, a sample of liver was collected from each animal for palmitoyl CoA oxidase activity analyses. Representative samples of liver, testes, and pancreas were collected from each animal and sent to Pathology Associates, A Charles River Company for proliferation cell nuclear antigen evaluation. A sample from 9 Covance 6329-230 3M T-6889.2 the liver and pancreas was collected from each animal and sent to the DuPont Haskell Company for possible receptor level determinations. Bile was collected from each animal and sent to the University of Dundee for bile acid determination. A sample of liver was collected from each animal and sent to 3M for APFO concentration analyses. Microscopic examinations were done on the adrenals, liver, pancreas, spleen, and testes from each animal. All animals survived to the scheduled sacrifice. There were no apparent test material-related effects on estradiol, estriol, thyroid stimulating hormone, total and free triiodothyronine, and total and free thyroxin. Estrone levels were notably lower for males given 2 and 20 mg/kg/day. There were no apparent test material-related effects on clinical observations, body weights, food consumption, clinical pathology parameters, or macroscopic or microscopic findings. There did not appear to be a cell proliferative response to the test material in the liver, testes or pancreas, as determined by proliferating indices which were similar in all animals. Based on the results of this study, daily capsule administration of Ammonium Perfluorooctanoate to cynomolgus monkeys for 29 days at dose levels of 2 and 20 mg/kg/day was well tolerated and produced no apparent evidence of toxicity. 10 PURPOSE Covance 6329-230 3M T-6889.2 The purpose of this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study. In addition, the effect on critical enzyme levels, hormones, and other selected biochemical parameters was determined. REGULATORY COMPLIANCE All aspects of this study were done in accordance with the Environmental Protection Agency Good Laboratory Practice Standards (GLPs), 40 CFR 792, except that bile acid determinations done by the University of Dundee were not done in compliance with GLPs. TEST MATERIAL Test Material The test material, Ammonium Perfluorooctanoate (APFO), Lot No. 332 (expiration date: December 15, 2001), is a white powder and is 95.2% pure. It was received at Covance on June 10, 1998. The test material was stored at room temperature. Information on synthesis methods, composition, or other characteristics that define the test material is on file with the Sponsor. The Certificate of Analysis is in Appendix 1. Reserve (Archive) Samples A reserve sample (1 g) of the test material was taken before initiation of treatment and stored at room temperature. This sample was transferred to the Sponsor on May 2, 2001. Disposition The remaining test material was returned on May 2, 2001. 11 TEST SYSTEM Covance 6329-230 3M T-6889.2 Test Animal Young adult to adult cynomolgus monkeys were obtained from Covance Research Products Inc. (Denver, Pennsylvania and Alice, Texas), on March 10, 1998. The animals weighed 2.1 to 3.6 kg at initiation of treatment. Identification Each animal was assigned a permanent number upon arrival and identified with collar tag before initiation of treatment. All data for an animal are recorded under this number. Acclimation Nine males were received on March 10, 1998, transferred from the Covance in-house stock colony on June 3, 1998. All animals were acclimated in Animal Room 2015; animals were acclimated at least 30 days before initiation of treatment (duration includes days animals were in the stock colony). In general, animals in this shipment appeared healthy. During acclimation, the animals were examined for abnormalities indicative of health problems. In addition, tuberculosis tests were done on each animal. Results of the tuberculosis tests were negative for all animals. One male was eliminated from study consideration because it had the highest body weight. There were eight males in the randomization for assignment to groups (two males in Group 1 and three males each in Groups 2 and 3). The animal not selected for the study was returned to the Covance stock colony. Housing and Maintenance Animal Room 2015 was used for this study. Environmental controls for the animal room were set to maintain 18 to 29oC, a relative humidity of 30 to 70%, and a 12-hour light/12-hour dark cycle. The animals were housed individually in stainless steel cages. Certified primate diet (#8726C, Harlan Teklad) was provided once or twice daily. The lot numbers are recorded in the data. The diet is routinely analyzed by the manufacturer 12 Covance 6329-230 3M T-6889.2 for nutritional components and environmental contaminants. Results of specified nutrient and contaminant analyses are on file with Covance-Madison. Fruit or additional supplements were provided, but did not require analysis. Water was provided ad libitum. Samples of the water are analyzed for specified microorganisms and environmental contaminants. The results are on file with Covance-Madison. There were no known contaminants in the diet or water at levels that would have interfered with this study. Justification APFO is a known hepatic peroxisome proliferator (PP) in the rat. When exposed to PP, nonhuman primates (such as the cynomolgus monkey) respond similarly to humans (i.e., low to no hepatic response) and therefore are an appropriate human surrogate species. PROCEDURES This study was conducted in accordance with the Protocol dated June 11, 1998, and Protocol Amendment No. 1 through 3. The protocol and protocol amendments are in Appendix 1. Group Designations and Dose Levels Selection of animals for the study was based on clinical observations, body weights, and other data as appropriate. Animals were assigned to treatment groups using a computerized blocking procedure designed to achieve body weight balance with respect to treatment group. 13 Covance 6329-230 3M T-6889.2 Dose Level Total Material Dose Level Group 1 (Control) (mg/kg/day) 0a (mg/kg/day) 0a Number of Males 2 2 (Low-dose) 2 2 3 3 (High-dose) 20 20 3 a The control group (Group 1) received the same size and number of empty gelatin capsules as used for Group 3. Dose Preparation Gelatin capsules (Torpac, Inc., Fairfield, New Jersey), Size No. 2, Lot Nos. 122630 (expiration date June 26, 2002) and 122932 (expiration date June 12, 2003), were used for dose administration. The test material was dispensed into capsules weekly. The dose levels were based on the test material as supplied. For Groups 2 and 3, the specified amount of test material was weighed and transferred into the gelatin capsules. The top and bottom halves of each capsule were joined, and the capsules were placed into the appropriate container labeled with study number, group number, animal number, and dose level. The prepared capsules were stored at room temperature. Dose Analyses Because the test material was not being mixed with a vehicle, dose analyses were not required. Method of Administration Gelatin capsules were used for test material administration to compare with previously conducted toxicology studies using the oral route. The dose preparations were administered orally in gelatin capsules once daily (7 days/week) for 29 days. The dosing tubes were flushed with 5 mL of reverse osmosis water. Individual doses were calculated based on the most recently recorded body weights, with the exception of body weight collection days when the previous body weight was used. Animals were dosed at approximately the same time each day. 14 Covance 6329-230 3M T-6889.2 Clinical Observations The animals were observed twice daily (a.m. and p.m.) for mortality and moribundity. Each animal was observed daily, and food consumption was assessed qualitatively; abnormal findings were recorded. Animals were observed approximately 30, 60, and 90 minutes postdose for signs of poor health or abnormal behavior. Effects were recorded as they were observed. Animals were observed once weekly; abnormal findings or an indication of normal was recorded. Body Weights Individual body weight data were recorded before initiation of treatment, on the first day of treatment, and weekly thereafter. An additional body weight was recorded on Day -1 for the Day 1 dose calculations. Blood Hormone Determination Blood was collected from a femoral vein of each animal before initiation of treatment (Day -8) and on Day 30 (after 29 days of treatment). Blood was collected between 07:00 and 09:00. Animals were fasted overnight before collections. Approximately 3 mL (Day -8) or 6 mL (Day 30) of blood for plasma samples were collected into tubes with potassium EDTA at the anticoagulant. Approximately 5 mL (Day -8) or 6 mL (Day 30) of blood for serum samples were collected without anticoagulant and allowed to clot for serum samples. Blood samples for plasma (Day 30) were maintained chilled until plasma was harvested. Blood samples collected on Day -8 for serum and plasma and blood samples collected on Day 30 for serum were maintained at room temperature until serum and plasma were harvested. Samples for serum were centrifuged within 1 hour after collection, and serum was harvested. Serum was divided into two approximately equal aliquots and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to Ani Lytics Inc., for analyses. The serum samples were analyzed for estradiol (E2), estrone (E1), estriol (E3), thyroid stimulating hormone (TSH), total and free triiodothyronine (T3 and FT3, respectively), and total and free thyroxin (T4 and FT4, respectively). Results of these analyses provided by Ani Lytics Inc., are in Appendix 6. Samples for plasma were centrifuged within 1 hour after collection. Plasma was harvested and stored in a freezer set to maintain -60 to -80C until packed on dry ice and 15 Covance 6329-230 3M T-6889.2 shipped to DuPont Haskell Company for possible future analyses. Results of the analyses for cholecystokinin, if any, will be reported separately. Serum APFO Level Determination Approximately 2 mL of whole blood were collected from a femoral vein of each animal before initiation of treatment (Day -8) and on Day 30 (after 29 days of treatment). Animals were fasted overnight before collections. All samples were collected without anticoagulant, maintained at room temperature, and allowed to clot. Samples were centrifuged within 1 hour after collection, serum was harvested and stored in a freezer set to maintain -10 to -30C until packed on dry ice and shipped to 3M for analyses. The samples were analyzed for APFO. Results of analyses will be reported separately by 3M. Clinical Pathology Blood samples were collected from each animal twice before initiation of treatment (Days -8 and -5) and on Day 30 (after 29 days of treatment) for hematology and clinical chemistry tests. In addition, blood for clinical chemistry tests was collected from each animal on Day 2 (approximately 24 hours after the first dose). Animals were fasted overnight; water was provided ad libitum. Blood was collected from a femoral vein. The following were evaluated: Hematology red blood cell (erythrocyte) count hemoglobin hematocrit mean corpuscular volume mean corpuscular hemoglobin mean corpuscular hemoglobin concentration platelet count white blood cell (leukocyte) count differential blood cell count segmented neutrophil count lymphocyte count monocyte count eosinophil count basophil count blood cell morphology reticulocyte count 16 glucose urea nitrogen creatinine total protein albumin globulin total bilirubin cholesterol triglycerides aspartate aminotransferase alanine aminotransferase alkaline phosphatase Covance 6329-230 3M T-6889.2 Clinical Chemistry gamma glutamyl transferase sorbitol dehydrogenase creatine kinase calcium inorganic phosphorus sodium potassium chloride bile acids amylase lipase pancreatic-specific amylase Additional Blood Collection Whole blood (approximately 15 mL) was collected from the vena cava of each animal at the time of exsanguination. Approximately equally sized samples (approximately 5 mL each) of serum (collected without anticoagulant) and whole blood and plasma using potassium EDTA as the anticoagulant were transferred into containers and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to 3M for possible future analysis. Necropsy On Day 30, animals that were fasted overnight were anesthetized with ketamine and xylazine, weighed, bled for the additional blood collection, exsanguinated, and necropsied. Animals were necropsied in random order. The necropsy included a macroscopic examination of the external surface of the body; all orifices; the cranial cavity; the brain and spinal cord; the nasal cavity and paranasal sinuses; cervical tissues and organs; and the thoracic, abdominal, and pelvic cavities and viscera. Palmitoyl CoA Oxidase Determinations Representative samples of the right lateral lobe of liver were collected from each animal at the scheduled sacrifice. The sample was weighed, flash-frozen in liquid nitrogen, and 17 Covance 6329-230 3M T-6889.2 stored in a freezer set to maintain -60 to -80oC until analyzed for palmitoyl CoA oxidase activity. Cell Proliferation Evaluation Representative samples of the left lateral lobe of the liver, left and right testes, and pancreas were collected and preserved in zinc formalin. After fixation, samples for proliferation cell nuclear antigen (PCNA) evaluation were embedded in paraffin and maintained at ambient temperature (with slides stained with hematoxylin and eosin) until shipped to Pathology Associates, A Charles River Company for PCNA analyses. Results of the evaluation provided by Pathology Associates, A Charles River Company are in Appendix 7. Receptor Level Determination Samples (approximately 2 g) of liver and pancreas was collected from each animal at the scheduled sacrifice, flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to the DuPont Haskell Company for possible analysis. Results of the receptor level determination, if any, will be reported separately. Bile Acid Determination Approximately 1.0 to 2.0 mL of bile were collected from each animal at the scheduled sacrifice, flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to the University of Dundee for bile acid determination. Results of the bile acid determination will be reported separately by the University of Dundee. Liver APFO Determination A section of liver (approximately 20 g) was collected from each animal at the scheduled sacrifice, weighed, flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped with plasma samples to 3M for APFO analysis. Results of these analysis will be reported separately by 3M. 18 Covance 6329-230 3M T-6889.2 Tissue Preservation The following tissues (when present) or representative samples were collected and preserved in 10% neutral-buffered formalin, unless otherwise specified, for possible future microscopic examination: adrenal (2) aorta brain cecum colon duodenum epididymis (2) esophagus eyes [preserved in Davidson's fixative(2)] femur with bone marrow (articular surface of the distal end) gallbladder heart ileum jejunum kidney (2) lesions liver lung lymph node (mesenteric) mammary gland muscle (thigh) pancreas pituitary prostate rectum salivary gland [mandibular (2)] sciatic nerve seminal vesicle (2) skin spinal cord (cervical, thoracic, and lumbar) spleen sternum with bone marrow stomach testis (2) thymus thyroid (2) with parathyroid trachea urinary bladder Histopathology The adrenals, liver, pancreas, spleen, and testes were embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically from each animal. Statistical Analyses Only data collected on or after the first day of treatment were analyzed statistically. One-way analysis of variance [ANOVA (Winer, 1971a)] was used to analyze initial body weights; palmitoyl CoA oxidase activities, and continuous clinical pathology values. Levene's test (Levene, 1960) was done to test for variance homogeneity. In the case of heterogeneity of variance at p < 0.05, transformations were used to stabilize the variance. 19 Covance 6329-230 3M T-6889.2 ANOVA was done on the homogeneous or transformed data. If the ANOVA was significant, Dunnett's t-test (Dunnett, 1964) was used for control versus treated group comparisons. One-way analysis of covariance [ANCOVA (Winer, 1971b)] was used to analyze body weights, with initial body weights as the covariate. Although Levene's test for variance homogeneity was done (see above), no transformations were used because covariance adjustment removed extraneous heterogeneity. If the ANCOVA was significant, least squares means t-test was used for control versus treated group comparisons. Groups 2 and 3 were compared with Group 1 (control). Group comparisons were evaluated at the 5.0%, two-tailed probability level. RECORD RETENTION All raw data, documentation, records, protocol, and specimens generated as a result of this study will be archived in the storage facilities of Covance-Madison for a period of 1 year. One year after the submission of the final report, the Sponsor will determine the final disposition of the materials. All raw data stored on magnetic media and an original copy of the final report will be retained by Covance-Madison. Within 1 year after submission of the final report, all the aforementioned materials from the Sponsor's designees (Ani Lytics Inc., DuPont Haskell Company, Pathology Associates International, and the University of Dundee) will be sent to the Sponsor (Paul Lieder, PhD, DABT, 3M). Pathology Associates, A Charles River Company (PAI) is responsible for maintenance of any raw data or specimens produced by PAI. RESULTS Clinical Observations Clinical observations are summarized in Tables 1 through 4; individual data are in Appendix 2. Individual animal fate data are also in Appendix 2. All animals survived to the scheduled sacrifice. There were no test material-related clinical observations. 20 Covance 6329-230 3M T-6889.2 On Day 23, Animal No. I05419 (Group 2) was observed with a 4-cm laceration on the inner left thigh. The animal was anesthetized with ketamine, and the wound was cleaned and sutured. This condition was not considered to be test material-related. Body Weights Body weight data are summarized in Table 5; individual data are in Appendix 3. There were no test material-related effects on body weights. Food Consumption Food consumption data are summarized in Table 1 (Summary of Clinical Observation - A.M./Weekly); individual data are included in the individual clinical observations in Appendix 2. Low or no food consumption was noted for one animal (Animal No. I05235) given 20 mg/kg/day. For this animal, no food consumption was noted on Day 12, and low food consumption was noted on Days 5, 7, 11, 14, 16, 17, and 24. The decrease in food consumption for this animal may be test material-related. Blood Hormone Analyses Summary and individual hormone analyses data are in Appendix 6. There were no apparent test material-related effects on estradiol, estriol, thyroid stimulating hormone, total and free triiodothyronine, and total and free thyroxin. Estrone levels were notably lower for males given 2 and 20 mg/kg/day. Clinical Pathology Hematology and clinical chemistry data are summarized in Tables 6 through 12; individual data are in Appendix 4. The Pathology Report contains a discussion of the data. Administration of APFO had no effects on clinical pathology test results. 21 Covance 6329-230 3M T-6889.2 Cell Proliferation Evaluation Results of cell proliferation evaluation provided by Pathology Associates, A Charles River Company are in Appendix 7. Cell proliferation, as determined by measuring the proliferating index, was not increased in the liver, testes or pancreas of monkeys. Thus, there did not appear to be a cell proliferative response to the test material in the liver, testes or pancreas, as determined by proliferating indices which were similar in all animals. Anatomical Pathology Incidences of macroscopic and microscopic observations are summarized in Tables 13 and 14. Individual data are in Appendix 5. The Pathology Report contains a discussion of the data. Administration of APFO had no effects on anatomical pathology test results. CONCLUSIONS Based on the results of this study, daily capsule administration of Ammonium Perfluorooctanoate to cynomolgus monkeys for 29 days at dose levels of 2 and 20 mg/kg/day was well tolerated and produced no apparent evidence of toxicity. 22 SIGNATURES Covance 6329-230 3M T-6889.2 23 Covance 6329-230 3M T-6889.2 REFERENCES Dunnett, C. W., "New Tables for Multiple Comparisons with a Control," Biometrics, 20:482-491 (1964). Levene, H., "Robust Tests for Equality of Variances," Contributions to Probability and Statistics, (eds.) I. Olkin et al., Ch. 25, pp. 278-292, Stanford University Press: Stanford, California (1960). Winer, B. J., "Design and Analysis of Single-Factor Experiments," Statistical Principles in Experimental Design, Second Ed., Ch. 3, pp. 149-260, McGraw-Hill: New York, New York (1971a). Winer, B. J., "Analysis of Covariance," Statistical Principles in Experimental Design, Second Ed., Ch. 10, pp. 752-812, McGraw-Hill: New York, New York (1971b). 24 PATHOLOGY REPORT Covance 6329-230 3M T-6889.2 SUMMARY The purpose of this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material, Ammonium Perfluorooctanoate (APFO), on critical enzyme levels, hormones, and other selected biochemical parameters. Administration of APFO had no effects on clinical or anatomical pathology test results. METHODS Three groups of male cynomolgus monkeys were administered the test material orally via capsule at a dose level of 0 (control group; received empty gelatin capsules), 2, or 20 mg/kg of body weight/day (mg/kg/day) for 29 days. There were two animals in the control group and three animals in each of the groups receiving APFO. All animals survived to the scheduled sacrifice. Blood was collected for hematology and clinical chemistry tests twice before initiation of treatment (Days -8 and -5) and on Day 30. In addition, blood for clinical chemistry tests was collected on Day 2. The animals were sacrificed and necropsied on Day 30; macroscopic observations were recorded, and tissues were preserved in fixative as specified by the protocol. Microscopic examinations were done on adrenals, liver, pancreas, spleen, and testes from all animals. Samples of liver were collected and frozen for determination of hepatic palmitoyl CoA oxidase activity and for determination of APFO content (done by 3M). Samples of liver, testes, and pancreas were collected and preserved in zinc formalin for evaluation of proliferation cell nuclear antigen (PCNA; done by Pathology Associates, a Charles River Company). Samples of liver and pancreas were collected and frozen for possible receptor level determination (done by the DuPont Haskell Company). Samples of bile were collected and frozen for bile acid determination (done by the University of Dundee). Statistically significant differences cited in the Results and Discussion section are based on comparisons between the control and treated groups. 25 RESULTS AND DISCUSSION Mortality All animals survived to the scheduled sacrifice. Covance 6329-230 3M T-6889.2 Clinical Pathology Individual values for hematology and clinical chemistry tests are in Appendix 4. Mean values and the results of statistical comparisons are in Tables 6 through 12. Days -8 and -5. Clinical pathology test results indicated no obvious group or individual health abnormalities. Day 2. Administration of the test material had no effects on clinical chemistry test results at this collection interval. There were only two statistically significant differences for clinical chemistry results between the control and treated groups; lower glucose for the low-dose group and lower total bilirubin for the high-dose group. These differences were considered incidental because similar differences were present before treatment was initiated. There were no other notable differences for clinical chemistry results between the control and treated groups. Day 30. Administration of the test material had no effects on hematology or clinical chemistry test results at Day 30. The only statistically significant differences between the control and treated groups was lower glucose for the high-dose males. This difference was considered incidental because a similar difference was present before treatment was initiated. There were no other notable differences for clinical pathology results between the control and treated groups. Anatomical Pathology Individual animal pathology data are in Appendix 5. Incidence summaries of the macroscopic and microscopic observations are in Tables 13 and 14, respectively. 26 Covance 6329-230 3M T-6889.2 Macroscopic Findings. There were no test material-related gross findings noted at necropsy. A few commonly occurring macroscopic findings frequently observed as spontaneous lesion in monkeys were present. Microscopic Findings. There were no test material-related microscopic findings. The only microscopic findings were minimal mineralization of the adrenals for one male given 2 mg/kg/day and immature testes for one male given 2 mg/kg/day and one male given 20 mg/kg/day. There were no differences in the appearances of the testes of the other two males given 2 mg/kg/day or the other two males given 20 mg/kg/day. 27 Covance 6329-230 3M T-6889.2 COMMENTS ON THE DATA Various models of calculators, computers, and computer programs were used to analyze data in this study. Because different models round off or truncate numbers differently, values in some tables (e.g., means, standard deviations, or individual values) may differ slightly from those in other tables, from individually calculated data, or from statistical analysis data. Neither the integrity nor the interpretation of the data was affected by these differences. Some tabular data were compiled using Excel Version 7.0 software. The summary tables for clinical observations indicate the number of animals for which a condition was observed without regard to the specific nature, severity, reversibility, number of incidences/animal, or the length of time the condition persisted. Only observations other than normal are indicated on the summary clinical observations tables. The day of initiation of treatment is "Day 1." Animal No. I05419 (Group 2) was observed and treated by the laboratory animal veterinary staff, and the data were recorded on the day the examination was done. These data are referenced in the Request for Veterinary Services and are archived with the raw data but do not appear on the summary or individual clinical observations tables. 28 CODES, ABBREVIATIONS, AND UNITS Covance 6329-230 3M T-6889.2 General Codes and Abbreviations Codes for Clinical Pathology Abbreviations and Units for Clinical Hematology Abbreviations and Units for Clinical Chemistry Codes for Anatomical Pathology Note: The following lists of codes, abbreviations, and units are used by Covance. Some, but not necessarily all, of this information may be needed for this report. 29 Covance 6329-230 3M T-6889.2 General Codes and Abbreviations WK N Mean; MEAN CAM SD; S.D.; STAND DEV; STANDARD DEV; sd * -; NA P UNSCHED DISPATCH MIN OBS Week. Number of measurements in a group. Arithmetic mean. Covariate-adjusted mean. Standard deviation. Group mean is significantly different from the mean of the control group (Group 1) at p < 0.05. No value; not applicable; not present. Present. Unscheduled. Observations transferred from the in-life module of the data collection system to the necropsy module for reference during necropsy. Observations are duplicates of the last in-life observations. Minute. A.M. daily/weekly observation. Animal Death Codes: T Terminal sacrifice. 30 NS QS/QNS NR FS SC SH H SL L SI I NF U DT/DOT DB TJ TE RE EE SE PC PD PI PL PA CO HB PLASMO NO AGG FR UTD NO COAG Covance 6329-230 3M T-6889.2 Codes for Clinical Pathology GENERAL CODES No sample Quantity not sufficient No repeat (sample volume not sufficient for repeat analysis) Fibrin strands Sample clotted Slightly hemolyzed Hemolyzed Slightly lipemic Lipemic Slightly icteric Icteric Animal not fasted Unscheduled/moribund bleed Animal died on test Died during bleeding Technician judgment to repeat test Technical error (instrument or technician error that results in unacceptable data, e.g., unacceptable instrument output, sample spilled, entry of invalid data) Recording error (recorded incorrect data, e.g., wrong number, spelling error, incorrect date) Entry error (incorrect keyboard entry) Sampling error Platelets clumped Platelets decreased Platelets increased Platelets large Platelets appear adequate Color interferes with test Heinz bodies observed Plasmodium No aggregation Fractious Unable to determine No coagulation 31 Covance 6329-230 3M T-6889.2 Codes for Clinical Pathology (Continued) RESULTS NOT INCLUDED IN STATISTICAL ANALYSES Hemolyzed clinical chemistry or coagulation samples Samples from animals at unscheduled intervals Prothrombin times (PT) greater than 50 seconds Activated partial thromboplastin times (PTT) greater than 110 seconds Bleed times (BLETIME) greater than 30 minutes CODES FOR BLOOD CELL MORPHOLOGY The following scale was used to measure the degree of anisocytosis (ANISO), poikilocytosis (POIK), polychromasia (POLY), hypochromasia (HYPO), and toxic neutrophils (TOXNEUT): Scale 1 2 3 4 Degree Normal for the species Slight Moderate Marked Not applicable Presence Not present Rare Few Moderate Many 32 Covance 6329-230 3M T-6889.2 Abbreviations and Units for Clinical Hematology Test Red blood cell count Hemoglobin Hematocrit Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet count Mean platelet volume Reticulocyte count Absolute reticulocyte count Heinz body count Erythrocyte sedimentation rate Prothrombin time Activated partial thromboplastin time Thrombin time Activated coagulation time Fibrinogen Fibrin/fibrinogen degradation products Platelet aggregation Collagen Adenosine diphosphate Alpha 2-antiplasmin Bleeding time Methemoglobin Plasma hemoglobin Myeloid/erythroid ratio Estimated myeloid/erythroid ratio White blood cell count Differential blood cell count Nucleated red blood cell count Corrected white blood cell count Segmented neutrophil count Band neutrophil count Lymphocyte count Monocyte count Eosinophil count Basophil count Anisocytosis Polychromasia Abbreviation (Units) RBC (E6/UL or X106/L) HGB (G/DL) HCT (%) MCV (FL) MCH (PG) MCHC (%) PLT (E3/UL or X103/L) MPV (FL) RETIC (%) RETIC (E3/UL or X103/L) HEINZ (%) ESR (MM/HR) PT (SEC) PTT (SEC) TT (SEC) ACT (SEC) FBR (MG/DL) FDP (UG/ML) PAGG/COL (%) PAGG/ADP (%) ANTIPLAS (%) BLE TIME (SEC) METHGB (%) PLA HGB (MG/DL) M/E RATIO EST M/E RATIO WBC (E3/UL or X103/L) NRBC (/100 WBC) COR WBC (E3/UL or X103/L) N-SEG (E3/UL or X103/L) and % N-BAND (E3/UL or X103/L) and % LYMPH (E3/UL or X103/L) and % MONO (E3/UL or X103/L) and % EOSIN (E3/UL or X103/L) and % BASO (E3/UL or X103/L) and % ANISO (-,1,2,3) POLY (-,1,2,3) 33 Covance 6329-230 3M T-6889.2 Abbreviations and Units for Clinical Hematology (Continued) Test Poikilocytosis Hypochromasia Howell-Jolly bodies Basophilic stippling Toxic neutrophils Atypical lymphocytes Aqueous white blood cell count (right eye) Aqueous white blood cell count (left eye) Abbreviation (Units) POIK (-,1,2,3) HYPO (-,1,2,3) HJBODY (-,1,2,3,4) BASTIP (-,1,2,3) TOXNEUT (-,1,2,3,4) ATYPLYM (-,1,2,3,4) R EYE (WBC/UL) L EYE (WBC/UL) 34 Covance 6329-230 3M T-6889.2 Abbreviations and Units for Clinical Chemistry Test Glucose Urea nitrogen Urea Creatinine Total protein Albumin Globulin Albumin/globulin ratio Total bilirubin Direct bilirubin Indirect bilirubin Cholesterol Triglyceride Urea nitrogen/creatinine ratio Total lipids Phospholipids High-density lipoprotein cholesterol Low-density lipoprotein cholesterol Uric acid Aspartate aminotransferase Alanine aminotransferase Alkaline phosphatase Gamma glutamyl transferase Sorbitol dehydrogenase Lactate dehydrogenase Creatine kinase Amylase Lipase Palmitoyl CoA oxidase Calcium Ionized calcium Inorganic phosphorus Sodium Potassium Chloride Magnesium Zinc Strontium Iron Abbreviation (Units) GLU (MG/DL) UN (MG/DL) UREA (MG/DL) CREAT (MG/DL) T PRO (G/DL) ALB (G/DL) GLOB (G/DL) A/G RATIO T BILI (MG/DL) D BILI (MG/DL) I BILI (MG/DL) CHOL (MG/DL) TRIG (MG/DL) UN/CREAT (RATIO) T LIPIDS (MG/DL) P LIPIDS (MG/DL) HDL (MG/DL) LDL (MG/DL) UA (MG/DL) AST/SGOT (IU/L) ALT/SGPT (IU/L) ALK PHOS (IU/L) GGT (IU/L) SDH (IU/L) LDH (IU/L) CK (IU/L) AMYLASE (IU/L) LIPASE (IU/L) PCOAO (IU/G) CA (MG/DL) ION CA (MG/DL) I PHOS (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) MG (MEQ/L or MG/DL) ZN (MG/L or PPM) SR (MG/L or PPM) FE (UG/DL) 35 Covance 6329-230 3M T-6889.2 Abbreviations and Units for Clinical Chemistry (Continued) Test Excess iron Total iron binding capacity Unbound iron binding capacity Percent iron saturation Plasma cholinesterase Red blood cell cholinesterase Brain cholinesterase Caudate putamen Hippocampus Frontal cortex Cerebellum Bicarbonate Serum hemoglobin Serum bile acids Fecal bile acids Average fecal weight Fecal bile acids (calculation) Osmolality Electrophoresis Albumin Alpha-1-globulin Alpha-2-globulin Beta globulin Gamma globulin High-density lipoprotein Low-density lipoprotein Very-low-density lipoprotein Insulin Adrenocorticotropic hormone Cortisol Glucagon Creatine kinase isoenzymes BB MB MM Pancreatic-specific amylase Abbreviation (Units) EX FE (UG/DL) TIBC (UG/DL) UIBC (UG/DL) FE %SAT (%) CHEP (MU/ML) CHER (MU/ML) CHEB (MU/ML) CAUD PUT (UMOL/G) HIPPOCAM (UMOL/G) F CORTEX (UMOL/G) CEREBELL (UMOL/G) BICARB (MMOL/L) SER HGB (MG/DL) SBA (UMOL/L or MG/DL) FBA (UG/ML) FCC WGT (G) FBA (MG/Day) OSMO (MOSM/KG) E ALB (G/DL) E A-1 (G/DL) E A-2 (G/DL) E BETA (G/DL) E GAMMA (G/DL) E-HDL (%) E-LDL (%) E-VLDL (%) INSULIN (UU/ML) ACTH (PG/ML) CORTISOL (UG/ML) GLUCAGON (PG/ML) CK-BB (U/L) CK-MB (U/L) CK-MM (U/L) P AMYL (U/L) 36 Covance 6329-230 3M T-6889.2 Codes for Anatomical Pathology MICROSCOPIC CODES Distribution of Findings Focal Diffuse Multifocal Grades for Severity or Amount 1 Minimal - the least amount of change that can be observed with the light microscope 2 Slight - less than average amount of change, but readily discernible as abnormal 3 Moderate - the average amount of change that is expected for a lesion 4 Moderately severe (marked) - a marked amount of change with possible loss of function of the affected cells or organs 5 Severe - a great amount of change with probable loss of function of the affected cell or organs and frequently involves large areas of the organ Abbreviation STOMACH, GL TISSUE ABBREVIATIONS Definition Glandular stomach 37 Table 1 Covance 6329-230 3M T-6889.2 Summary of Clinical Observations A.M./Weekly 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ NUMBER OF ANIMALS AFFECTED DAYS 1-30 SEX: -----MALE---- GROUP: 1 2 3 CATEGORY DOSE: 0 2 20 KEYWORD QUALIFIER NUMBER: 2 3 3 ------------------------------------------------------------------------------------------------------------------------------- *** TOP OF LIST *** EXCRETION NO FECES 001 SKIN & PELAGE BROKEN SKIN THIGH-LEFT SCAB(S) LIMB-HIND-LEFT 010 010 QUALITATIVE FOOD CONSUMPTION LOW NONE *** END OF LIST *** 111 001 38 Table 2 Covance 6329-230 3M T-6889.2 Summary of Clinical Observations 30 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ NUMBER OF ANIMALS AFFECTED DAYS 1-29 SEX: -----MALE---- GROUP: 1 2 3 CATEGORY DOSE: 0 2 20 KEYWORD QUALIFIER NUMBER: 2 3 3 ------------------------------------------------------------------------------------------------------------------------------ *** TOP OF LIST *** SKIN & PELAGE BROKEN SKIN DIGIT(S)-HIND-RIGHT 100 THIGH-LEFT 010 SCAB(S) LIMB-HIND-LEFT 010 *** END OF LIST *** 39 Table 3 Covance 6329-230 3M T-6889.2 Summary of Clinical Observations 60 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ NUMBER OF ANIMALS AFFECTED DAYS 1-29 SEX: -----MALE---- GROUP: 1 2 3 CATEGORY DOSE: 0 2 20 KEYWORD QUALIFIER NUMBER: 2 3 3 ------------------------------------------------------------------------------------------------------------------------------ *** TOP OF LIST *** SKIN & PELAGE BROKEN SKIN DIGIT(S)-HIND-RIGHT 100 THIGH-LEFT 010 SCAB(S) LIMB-HIND-LEFT 010 *** END OF LIST *** 40 Table 4 Covance 6329-230 3M T-6889.2 Summary of Clinical Observations 90 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ NUMBER OF ANIMALS AFFECTED DAYS 1-29 SEX: -----MALE---- GROUP: 1 2 3 CATEGORY DOSE: 0 2 20 KEYWORD QUALIFIER NUMBER: 2 3 3 ------------------------------------------------------------------------------------------------------------------------------ *** TOP OF LIST *** APPEARANCE UNDER EFFECTS OF ANESTHESIA 010 SKIN & PELAGE BROKEN SKIN DIGIT(S)-HIND-RIGHT THIGH-LEFT SCAB(S) LIMB-HIND-LEFT *** END OF LIST *** 100 010 010 41 Table 5 Covance 6329-230 3M T-6889.2 Summary of Body Weight Data (kg) 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ SEX: ---------------MALE--------------- GROUP: 1 2 3 DOSE: 0 2 20 WEEK UNITS: MG/KG/DAY MG/KG/DAY MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------ -2 N MEAN S.D. a -1 N MEAN S.D. b -1 N MEAN S.D. 2 3.0 0.71 2 3.0 0.71 2 3.1 0.78 3 2.8 0.70 3 2.9 0.75 3 2.8 0.75 3 2.7 0.50 3 2.8 0.50 3 2.8 0.50 1 N MEAN S.D. 2 3.1 0.71 3 2.9 0.75 3 2.8 0.55 2 N CAM MEAN S.D. 2 2.9 3.1 0.57 3 2.9 2.9 0.70 3 2.9 2.8 0.56 3 N CAM MEAN S.D. 2 2.9 3.2 0.64 3 3.0 2.9 0.70 3 2.9 2.8 0.56 4 N CAM MEAN S.D. 2 2.9 3.1 0.64 3 2.9 2.9 0.70 3 2.9 2.8 0.50 5 N CAM MEAN S.D. 2 2.9 3.2 0.64 3 3.0 3.0 0.75 3 3.0 2.9 0.56 a Day -7. b Day -1. 42 Table 6 Summary of Clinical Hematology Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X106/L G/DL % FL PG % X103/L % X103/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 5.96 12.6 40.5 70.4 22.2 31.3 430 .4 S.D. 2.157 .92 6.65 14.28 6.43 2.83 75.7 .42 N 2 2 2 2 2 2 2 2 19 17.0 2 2 MEAN 6.85 13.6 44.2 65.8 20.3 30.7 422 .6 S.D. 1.196 .57 1.21 11.04 4.63 1.80 41.1 .57 N 3 3 3 3 3 3 3 3 34 30.0 3 20 MEAN 6.39 13.3 42.3 67.2 21.2 31.4 522 .3 S.D. .925 .96 .72 10.84 4.78 2.22 131.7 .23 N 3 3 3 3 3 3 3 3 20 10.7 3 43 Table 6 Summary of Clinical Hematology Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day ------ 0 MEAN S.D. N 2 MEAN S.D. N 20 MEAN S.D. N WBC X103/L -------- 10.4 1.63 2 9.4 1.01 3 9.6 1.10 3 N-SEG X103/L -------- 1.8 1.13 2 2.6 1.51 3 2.8 1.83 3 LYMPH X103/L -------- 7.4 .64 2 5.9 1.40 3 6.0 1.38 3 MONO X103/L -------- .6 .28 2 .9 .42 3 .5 .15 3 EOSIN X103/L -------- .6 .71 2 .1 .06 3 .3 .15 3 BASO X103/L -------- .0 .00 2 .0 .00 3 .0 .06 3 N-SEG% LYMPH% MONO% EOSIN% BASO% -------- -------- -------- -------- -------- 17 72 6 6 0 8.5 17.0 2.1 6.4 .0 2 2 2 2 2 27 63 9 1 0 15.5 18.1 3.2 .6 .0 3 3 3 3 3 28 63 6 2 0 16.6 18.1 1.2 1.5 .6 3 3 3 3 3 44 Table 7 Summary of Clinical Hematology Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X106/L G/DL % FL PG % X103/L % X103/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 5.47 11.5 37.3 70.6 22.0 31.0 404 .6 S.D. 1.711 .35 4.45 14.00 6.29 2.83 61.5 .85 N 2 2 2 2 2 2 2 2 26 36.1 2 2 MEAN 5.91 11.8 38.4 66.3 20.6 30.8 395 .9 S.D. 1.061 .49 1.17 10.81 4.93 2.22 22.1 .61 N 3 3 3 3 3 3 3 3 51 24.5 3 20 MEAN 5.75 11.9 37.9 67.3 21.2 31.4 500 .2 9 S.D. 1.057 .42 1.48 10.68 4.83 2.20 101.4 .12 4.4 N 3 3 3 3 3 3 3 3 3 45 Table 7 Summary of Clinical Hematology Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day ------ 0 MEAN S.D. N 2 MEAN S.D. N 20 MEAN S.D. N WBC X103/L -------- 11.1 1.56 2 10.7 3.18 3 10.5 3.10 3 N-SEG X103/L -------- 2.0 1.13 2 4.3 1.81 3 4.1 1.89 3 LYMPH X103/L -------- 7.8 .49 2 5.6 1.76 3 5.4 .76 3 MONO X103/L -------- .7 .14 2 .7 .12 3 .6 .32 3 EOSIN X103/L -------- .6 .64 2 .2 .10 3 .2 .21 3 BASO X103/L -------- .0 .07 2 .0 .00 3 .0 .00 3 N-SEG% LYMPH% MONO% EOSIN% BASO% -------- -------- -------- -------- -------- 18 72 6 4 0 7.8 14.1 .7 4.9 .7 2 2 2 2 2 40 52 6 2 0 12.1 12.6 .6 .6 .0 3 3 3 3 3 38 54 5 2 0 9.0 12.7 2.1 1.7 .0 3 3 3 3 3 46 Table 8 Summary of Clinical Hematology Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X106/L G/DL % FL PG % X103/L % X103/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 5.96 12.4 40.8 70.1 21.6 30.5 461 .2 S.D. 1.457 .14 2.26 13.44 5.52 2.05 66.5 .21 N 2 2 2 2 2 2 2 2 14 9.2 2 2 MEAN 6.74 13.6 44.3 66.5 20.5 30.6 407 .2 S.D. .824 1.19 1.51 11.02 4.52 1.63 4.2 .21 N 3 3 3 3 3 3 3 3 10 11.8 3 20 MEAN 6.24 13.0 41.9 68.1 21.3 31.1 489 .2 S.D. .814 1.11 1.31 10.91 4.78 2.25 116.0 .21 N 3 3 3 3 3 3 3 3 14 12.1 3 47 Table 8 Summary of Clinical Hematology Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day ------ 0 MEAN S.D. N 2 MEAN S.D. N 20 MEAN S.D. N WBC X103/L -------- 9.8 .07 2 8.0 2.29 3 8.3 .85 3 N-SEG X103/L -------- 1.6 .64 2 2.2 .79 3 3.1 1.11 3 LYMPH X103/L -------- 7.2 .78 2 5.4 1.60 3 4.4 1.31 3 MONO X103/L -------- .6 .21 2 .3 .15 3 .6 .10 3 EOSIN X103/L -------- .4 .35 2 .0 .06 3 .1 .12 3 BASO X103/L -------- .0 .07 2 .0 .00 3 .0 .00 3 N-SEG% LYMPH% MONO% EOSIN% BASO% -------- -------- -------- -------- -------- 16 74 6 4 0 7.1 7.8 2.1 3.5 .7 2 2 2 2 2 27 67 5 1 0 4.6 6.6 2.9 .0 .0 3 3 3 3 3 38 53 7 2 1 12.7 14.6 1.5 .6 .6 3 3 3 3 3 48 Table 9 Summary of Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL TRIG MG/DL ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 74 22 1.0 8.7 5.2 3.5 .4 S.D. 20.5 7.8 .21 2.40 1.13 1.27 .14 N 2 2 2 2 2 2 2 9 146 5.7 9.2 2 2 34 7.8 2 2 MEAN 76 18 1.1 8.4 5.1 3.3 .3 S.D. 14.2 2.0 .26 1.30 .30 1.00 .12 N 3 3 3 3 3 3 3 9 174 2.5 19.9 3 3 36 6.7 3 20 MEAN 64 23 1.2 8.0 4.8 3.3 .2 12 141 31 S.D. 5.1 2.3 .25 .55 .21 .75 .06 .6 12.7 6.7 N 3 3 3 3 3 3 3 3 3 3 49 Table 9 Summary of Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE IU/L LIPASE IU/L P AMYL U/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN S.D. N 41 1.4 2 45 9.9 2 847 420.0 2 122 11.3 2 2 348 492 .0 244.0 70.0 2 2 2 43 1.4 2 252 33.2 2 2 MEAN S.D. N 48 14.7 3 53 22.2 3 1013 106.1 3 159 45.1 3 5 369 522 4.4 159.3 174.5 3 3 3 46 23.4 3 250 41.7 3 20 MEAN S.D. N 52 6.7 3 59 26.7 3 948 65.5 3 136 35.5 3 5 195 496 1.2 40.3 32.0 3 3 3 43 8.5 3 239 37.3 3 50 Table 9 Summary of Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L ------ -------- -------- -------- -------- -------- 0 MEAN 11.4 S.D. 1.70 N 2 7.2 .99 2 158 11.3 2 6.6 .71 2 112 1.4 2 2 MEAN 11.0 S.D. .35 N 3 7.5 1.11 3 158 4.6 3 5.6 .35 3 108 1.2 3 20 MEAN S.D. N 10.9 .53 3 7.0 .29 3 154 1.5 3 6.1 .40 3 108 .6 3 Covance 6329-230 3M T-6889.2 51 Table 10 Summary of Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL TRIG MG/DL ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 84 18 1.2 8.4 5.0 3.4 .6 S.D. 41.0 9.2 .14 1.70 .78 .92 .07 N 2 2 2 2 2 2 2 4 138 2.1 26.2 2 2 28 .7 2 2 MEAN 66 15 1.1 8.0 4.8 3.2 .3 S.D. 6.5 1.0 .21 .85 .10 .90 .17 N 3 3 3 3 3 3 3 5 155 4.0 12.3 3 3 34 3.8 3 20 MEAN 67 20 1.1 8.0 4.7 3.4 .4 S.D. 14.4 5.1 .15 .57 .25 .75 .31 N 3 3 3 3 3 3 3 1 141 1.0 13.5 3 3 39 11.0 3 52 Table 10 Summary of Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE IU/L LIPASE IU/L P AMYL U/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN S.D. N 38 4.9 2 43 12.7 2 788 393.9 2 112 6.4 2 3 494 495 .0 478.7 76.4 2 2 2 40 2.8 2 252 26.2 2 2 MEAN S.D. N 39 13.3 3 49 15.2 3 930 83.5 3 137 39.7 3 3 279 520 1.0 227.6 174.8 3 3 3 56 9.2 3 252 37.2 3 20 MEAN S.D. N 48 9.8 3 51 14.5 3 896 54.7 3 121 38.2 3 3 171 466 1.0 29.3 15.0 3 3 3 57 13.5 3 229 18.4 3 53 Table 10 Summary of Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L ------ -------- -------- -------- -------- -------- 0 MEAN 10.9 S.D. 1.27 N 2 6.8 1.27 2 156 9.2 2 5.4 .92 2 108 2.1 2 2 MEAN 10.4 S.D. .61 N 3 6.4 1.04 3 155 3.5 3 5.2 .75 3 110 2.6 3 20 MEAN S.D. N 10.4 .51 3 6.2 .17 3 152 2.6 3 5.0 .58 3 107 1.7 3 Covance 6329-230 3M T-6889.2 54 Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL TRIG MG/DL ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 76 21 1.0 9.0 5.2 3.9 .4 10 152 28 S.D. 4.2 7.1 .21 2.19 .92 1.27 .21 4.9 7.8 7.8 N 2 2 2 2 2 2 2 2 2 2 2 MEAN 65 * 20 1.0 8.4 4.9 3.5 .3 S.D. 4.2 4.9 .21 .65 .10 .75 .06 N 3 3 3 3 3 3 3 9 169 3.0 22.3 3 3 52 8.3 3 20 MEAN S.D. N 68 2.0 3 23 1.0 3 1.0 .12 3 8.3 .51 3 4.8 .21 3 3.5 .70 3 .1 * .06 3 14 4.4 3 132 10.0 3 45 11.1 3 55 Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE IU/L LIPASE IU/L P AMYL U/L ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN S.D. N 47 2.8 2 53 25.5 2 872 355.7 2 125 5.7 2 1 225 477 .0 62.2 59.4 2 2 2 29 1.4 2 249 29.7 2 2 MEAN S.D. N 53 32.7 3 52 26.8 3 1029 113.3 3 148 44.9 3 2 794 1.5 1173.2 3 3 483 144.3 3 44 7.5 3 244 42.7 3 20 MEAN S.D. N 55 4.9 3 56 22.4 3 921 124.2 3 126 39.1 3 1 165 479 1.0 44.4 40.1 3 3 3 33 12.2 3 234 32.5 3 56 Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L ------ -------- -------- -------- -------- -------- 0 MEAN 11.6 S.D. 1.63 N 2 8.8 .28 2 162 12.0 2 6.2 .99 2 110 4.9 2 2 MEAN 10.5 S.D. .57 N 3 8.0 1.30 3 155 2.5 3 5.3 .44 3 109 3.8 3 20 MEAN S.D. N 10.7 .83 3 7.4 .90 3 156 4.2 3 5.5 .67 3 109 1.5 3 Covance 6329-230 3M T-6889.2 57 Table 12 Summary of Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE mg/kg/day GLU MG/DL UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL TRIG MG/DL ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 84 19 1.0 8.6 5.1 3.4 .5 S.D. 9.2 5.7 .07 1.77 .71 1.06 .28 N 2 2 2 2 2 2 2 4 150 3.5 31.8 2 2 32 .7 2 2 MEAN 76 17 1.1 8.3 5.0 3.3 .1 S.D. 3.2 2.1 .06 .60 .17 .75 .06 N 3 3 3 3 3 3 3 4 147 1.5 9.2 3 3 43 3.2 3 20 MEAN 65 * 20 1.2 7.8 4.5 3.3 .1 S.D. 2.1 3.5 .35 .47 .23 .70 .15 N 3 3 3 3 3 3 3 6 138 4.5 31.5 3 3 52 10.6 3 58 Table 12 Summary of Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE IU/L LIPASE IU/L P AMYL U/L PCOAO IU/G ------ -------- -------- -------- -------- -------- -------- -------- -------- -------- -------- 0 MEAN 52 59 917 110 2 511 471 42 242 2 S.D. .7 24.0 335.2 11.3 1.4 446.9 28.3 16.3 6.4 .7 N 2 2 2 2 2 2 2 2 2 2 2 MEAN 50 75 1111 136 2 324 500 38 244 1 S.D. 19.7 33.3 88.9 35.8 1.0 220.4 155.0 7.8 49.9 1.7 N 3 3 3 3 3 3 3 3 3 3 20 MEAN 55 83 704 101 2 207 441 39 218 6 S.D. 11.5 50.0 87.8 33.2 1.5 9.0 71.8 11.6 52.8 3.1 N 3 3 3 3 3 3 3 3 3 3 59 Table 12 Summary of Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L ------ -------- -------- -------- -------- -------- 0 MEAN 10.7 S.D. .71 N 2 7.2 1.77 2 153 5.7 2 5.4 .49 2 106 2.8 2 2 MEAN 10.8 S.D. .53 N 3 6.7 1.06 3 155 2.1 3 5.2 .90 3 109 2.9 3 20 MEAN S.D. N 10.2 .70 3 5.5 .31 3 151 5.1 3 5.2 .66 3 109 2.1 3 Covance 6329-230 3M T-6889.2 60 TABLE 13 Incidence of Macroscopic Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH=ALL;SUBSET=ALL --- N U M B E R - O F - A N I M A L S - A F F E C T E D SEX: -----MALE---GROUP: -1- -2- -3- ORGAN AND KEYWORD(S) OR PHRASE NUMBER: 2 3 3 ------------------------------------------------------------------------ -=- -=- -=- ** TOP OF LIST ** GENERAL COMMENT (GC) ................................. NUMBER EXAMINED: 2 3 3 EYES - DAVIDSONS NO MACROSCOPIC LESIONS 233 031 KIDNEY (KD) .......................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 1 3 3 LIGHT FOCUS(I)/AREA(S) 100 LUNG (LU) ............................................ NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 3 2 ADHESION(S) 001 STOMACH, GL (ST) ..................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 1 3 1 RED FOCUS(I)/AREA(S) ** END OF LIST ** 102 61 TABLE 14 Incidence of Microscopic Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH=ALL;FIND=ALL;SUBSET=ALL --- N U M B E R - O F - A N I M A L S - A F F E C T E D SEX: -----MALE---GROUP: -1- -2- -3- ORGAN AND FINDING DESCRIPTION NUMBER: 2 3 3 ------------------------------------------------------------------------ -=- -=- -=- ** TOP OF LIST ** LIVER (LI) ........................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 3 3 SPLEEN (SP) .......................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 3 3 PANCREAS (PA) ........................................ NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 3 3 TESTIS (TE) .......................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 2 2 --IMMATURE 011 ADRENAL (AD) ......................................... NUMBER EXAMINED: 2 3 3 NOT REMARKABLE: 2 2 3 --MINERALIZATION 010 ** END OF LIST ** 62 APPENDIX 1 Protocol Protocol Amendment No. 1 Protocol Amendment No. 2 Protocol Amendment No. 3 Material Safety Data Sheet Certificate of Analysis Covance 6329-230 3M T-6889.2 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 Covance 6329-230 3M T-6889.2 91 Covance 6329-230 3M T-6889.2 92 Covance 6329-230 3M T-6889.2 93 Covance 6329-230 3M T-6889.2 94 Covance 6329-230 3M T-6889.2 95 Covance 6329-230 3M T-6889.2 96 Covance 6329-230 3M T-6889.2 97 Covance 6329-230 3M T-6889.2 98 Covance 6329-230 3M T-6889.2 99 APPENDIX 2 Individual Animal Fate Data Individual Clinical Observations Covance 6329-230 3M T-6889.2 100 APPENDIX 2 Individual Animal Fate Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER -----I05353 I05417 I05231 I05352 I05419 I05235 I05351 I05416 DOSE GROUP ----- 1 1 2 2 2 3 3 3 SEX ------ MALE MALE MALE MALE MALE MALE MALE MALE DEATH CODE ----- T T T T T T T T TYPE OF DEATH ----------SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED DESCRIPTION OF DEATH ------------------------TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE DATE OF DEATH -------07/16/98 07/16/98 07/16/98 07/16/98 07/16/98 07/16/98 07/16/98 07/16/98 DAY OF STUDY ------30 30 30 30 30 30 30 30 WEEK OF STUDY ------5 5 5 5 5 5 5 5 101 Appendix 2 Individual Clinical Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: M1 DOSE: 0 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP ANIMAL DEATH WK OF KEYWORD OBS 30 MIN 60 MIN 90 MIN DISPATCH NUMBER CODE DEATH QUALIFIER DAYS 1-30 DAY =============================================================================================================================== I05353 T 5 SKIN & PELAGE BROKEN SKIN DIGIT(S)-HIND-RIGHT 18 - P P P - NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P I05417 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P QUALITATIVE FOOD CONSUMPTION LOW 21 P - - - - 102 Appendix 2 Individual Clinical Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 2 ------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: M2 DOSE: 2 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP ANIMAL DEATH WK OF KEYWORD OBS 30 MIN 60 MIN 90 MIN DISPATCH NUMBER CODE DEATH QUALIFIER DAYS 1-30 DAY ============================================================================================================================== I05231 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P I05352 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P I05419 T 5 APPEARANCE UNDER EFFECTS OF ANESTHESIA 23 - - - P - SKIN & PELAGE BROKEN SKIN THIGH-LEFT SCAB(S) LIMB-HIND-LEFT 24 P P P P - 25 - P P P - 26 P P P P - 27 P P P P - 28 P P P P - 29 P - - - - 30 P - - - P NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 103 Appendix 2 Individual Clinical Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 3 ------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: M2 DOSE: 2 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP ANIMAL DEATH WK OF KEYWORD OBS 30 MIN 60 MIN 90 MIN DISPATCH NUMBER CODE DEATH QUALIFIER DAYS 1-30 DAY =============================================================================================================================== (CONTINUED FROM PREVIOUS PAGE) I05419 T 5 NORMAL NO REMARKABLE OBSERVATIONS 8 P - - - - 15 P - - - - 22 P - - - - QUALITATIVE FOOD CONSUMPTION LOW 24 P - - - - 104 Appendix 2 Individual Clinical Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 4 ------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: M3 DOSE: 20 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP ANIMAL DEATH WK OF KEYWORD OBS 30 MIN 60 MIN 90 MIN DISPATCH NUMBER CODE DEATH QUALIFIER DAYS 1-30 DAY ============================================================================================================================== I05235 T 5 EXCRETION NO FECES 17 P - - - - NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P QUALITATIVE FOOD CONSUMPTION LOW NONE 5 P - - - - 7 P - - - - 11 P - - - - 14 P - - - - 16 P - - - - 17 P - - - - 24 P - - - - 12 P - - - - I05351 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P 105 Appendix 2 Individual Clinical Observations Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 5 ------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: M3 DOSE: 20 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP ANIMAL DEATH WK OF KEYWORD OBS 30 MIN 60 MIN 90 MIN DISPATCH NUMBER CODE DEATH QUALIFIER DAYS 1-30 DAY ============================================================================================================================== I05416 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1 P - - - - 8 P - - - - 15 P - - - - 22 P - - - - 29 P - - - - 30 P - - - P 106 APPENDIX 3 Individual Body Weight Data (kg) Covance 6329-230 3M T-6889.2 107 Appendix 3 Individual Body Weight Data (kg) Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL WEEK a WEEK b WEEK WEEK WEEK WEEK WEEK WEEK NUMBER -2 -1 -1 1 2 3 4 5 ------------------------------------------------------------------------------------------------------------------------------ GROUP: MALE 1 - 0 MG/KG/DAY I05353 I05417 2.5 2.5 2.5 2.6 2.7 2.7 2.6 2.7 3.5 3.5 3.6 3.6 3.5 3.6 3.5 3.6 GROUP: MALE 2 - 2 MG/KG/DAY I05231 I05352 I05419 2.7 2.9 2.8 2.9 2.9 3.0 2.9 3.0 2.1 2.1 2.1 2.1 2.2 2.2 2.2 2.2 3.5 3.6 3.6 3.6 3.6 3.6 3.6 3.7 GROUP: MALE 3 - 20 MG/KG/DAY I05235 I05351 I05416 2.7 2.8 2.7 2.8 2.7 2.7 2.8 2.8 2.2 2.3 2.3 2.2 2.3 2.3 2.3 2.4 3.2 3.3 3.3 3.3 3.4 3.4 3.3 3.5 a Day -7. b Day -1. 108 APPENDIX 4 Individual Clinical Hematology Data Individual Clinical Chemistry Data Covance 6329-230 3M T-6889.2 109 ANIMAL NUMBER -----Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Individual Clinical Hematology Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS RBC X106/L HGB G/DL -------- -------- Dose Level: 0 HCT MCV MCH % FL PG -------- -------- -------- Dosage Unit: mg/kg/day MCHC % -------- PLT X103/L -------- RETIC % -------- RETIC X103/L -------- 4.44 11.9 35.8 80.5 26.8 33.3 376 .7 31 7.49 13.2 45.2 60.3 17.7 29.3 483 .1 7 5.96 12.6 40.5 70.4 22.2 31.3 430 .4 2.157 .92 6.65 14.28 6.43 2.83 75.7 .42 2 2 2 2 2 2 2 2 19 17.0 2 Dose Level: 2 Dosage Unit: mg/kg/day 7.08 13.1 43.5 61.5 18.5 30.0 455 .4 28 5.56 14.2 43.5 78.3 25.6 32.7 376 1.2 67 7.92 13.4 45.6 57.5 16.9 29.3 435 .1 8 6.85 13.6 44.2 65.8 20.3 30.7 422 .6 1.196 .57 1.21 11.04 4.63 1.80 41.1 .57 3 3 3 3 3 3 3 3 34 30.0 3 Dose Level: 20 Dosage Unit: mg/kg/day 7.18 12.4 42.7 59.4 17.2 29.0 530 .2 14 5.37 14.3 42.8 79.6 26.5 33.4 649 .6 32 6.61 13.1 41.5 62.7 19.9 31.7 386 .2 13 6.39 13.3 42.3 67.2 21.2 31.4 522 .3 .925 .96 .72 10.84 4.78 2.22 131.7 .23 3 3 3 3 3 3 3 3 20 10.7 3 Covance 6329-230 3M T-6889.2 110 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Covance 6329-230 3M T-6889.2 Individual Clinical Hematology Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS WBC X103/L -------- N-SEG X103/L -------- LYMPH X103/L -------- MONO X103/L -------- EOSIN X103/L -------- BASO X103/L -------- N-SEG% -------- LYMPH% -------- MONO% -------- EOSIN% -------- BASO% -------- Dose Level: 0 Dosage Unit: mg/kg/day 9.2 1.0 7.8 .4 .1 .0 11 84 11.5 2.6 6.9 .8 1.1 .0 23 60 4 1 0 7 10 0 10.4 1.8 7.4 .6 .6 .0 17 72 6 6 0 1.63 1.13 .64 .28 .71 .00 8.5 17.0 2.1 6.4 .0 2 2 2 2 2 2 2 2 2 2 2 Dose Level: 2 Dosage Unit: mg/kg/day 9.3 4.0 4.3 1.0 .1 .0 43 46 10 8.5 1.0 7.0 .4 .1 .0 12 82 5 10.5 2.8 6.3 1.2 .2 .0 27 60 11 1 0 1 0 2 0 9.4 2.6 5.9 .9 .1 .0 27 63 9 1 0 1.01 1.51 1.40 .42 .06 .00 15.5 18.1 3.2 .6 .0 3 3 3 3 3 3 3 3 3 3 3 Dose Level: 20 Dosage Unit: mg/kg/day 9.0 3.2 5.0 .4 .4 .0 35 56 5 4 0 9.0 .8 7.6 .5 .1 .1 9 84 5 1 1 10.9 4.4 5.5 .7 .3 .0 40 50 7 2 0 9.6 2.8 6.0 .5 .3 .0 28 63 6 2 0 1.10 1.83 1.38 .15 .15 .06 16.6 18.1 1.2 1.5 .6 3 3 3 3 3 3 3 3 3 3 3 111 Appendix 4 Individual Clinical Hematology Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ ANISO -------- POLY -------- POIK -------- HYPO -------- TOXNEUT -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 - - - - - I05417 - - - - - Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 - - - - - I05352 - - - - - I05419 - - - - - Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 - - - - - I05351 - - - - - I05416 - - - - - Covance 6329-230 3M T-6889.2 112 Appendix 4 Individual Clinical Hematology Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ RBC HGB HCT X106/L G/DL % -------- -------- -------- MCV FL -------- MCH PG -------- MCHC % -------- PLT X103/L RETIC % RETIC X103/L -------- -------- -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 I05417 4.26 11.3 34.2 80.5 26.5 33.0 360 1.2 51 6.68 11.8 40.5 60.7 17.6 29.0 447 .0 0 MEAN S.D. N 5.47 11.5 37.3 70.6 22.0 31.0 404 .6 1.711 .35 4.45 14.00 6.29 2.83 61.5 .85 2 2 2 2 2 2 2 2 26 36.1 2 Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 I05352 I05419 6.27 11.5 38.9 62.0 18.3 29.5 418 .8 50 4.72 12.4 37.1 78.6 26.3 33.4 393 1.6 76 6.75 11.6 39.3 58.3 17.3 29.6 374 .4 27 MEAN S.D. N 5.91 11.8 38.4 66.3 20.6 30.8 395 .9 1.061 .49 1.17 10.81 4.93 2.22 22.1 .61 3 3 3 3 3 3 3 3 51 24.5 3 Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 I05351 I05416 6.61 11.4 39.2 59.3 17.3 29.2 516 .1 7 4.57 12.2 36.3 79.4 26.6 33.6 593 .3 14 6.07 12.0 38.3 63.1 19.7 31.3 392 .1 6 MEAN S.D. N 5.75 11.9 37.9 67.3 21.2 31.4 500 .2 9 1.057 .42 1.48 10.68 4.83 2.20 101.4 .12 4.4 3 3 3 3 3 3 3 3 3 Covance 6329-230 3M T-6889.2 113 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Covance 6329-230 3M T-6889.2 Appendix 4 Individual Clinical Hematology Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS WBC X103/L -------- N-SEG X103/L -------- LYMPH X103/L -------- MONO X103/L -------- EOSIN X103/L -------- BASO X103/L -------- N-SEG% -------- LYMPH% -------- MONO% -------- EOSIN% -------- BASO% -------- Dose Level: 0 Dosage Unit: mg/kg/day 10.0 1.2 8.2 .6 .1 .1 12 82 6 1 1 12.2 2.8 7.5 .8 1.0 .0 23 62 7 8 0 11.1 2.0 7.8 .7 .6 .0 18 72 6 4 0 1.56 1.13 .49 .14 .64 .07 7.8 14.1 .7 4.9 .7 2 2 2 2 2 2 2 2 2 2 2 Dose Level: 2 Dosage Unit: mg/kg/day 9.0 4.6 3.7 .6 .2 .0 51 40 7 2 0 8.8 2.4 5.8 .6 .1 .0 27 65 6 1 0 14.4 6.0 7.2 .8 .3 .0 42 50 6 2 0 10.7 4.3 5.6 .7 .2 .0 40 52 6 2 0 3.18 1.81 1.76 .12 .10 .00 12.1 12.6 .6 .6 .0 3 3 3 3 3 3 3 3 3 3 3 Dose Level: 20 Dosage Unit: mg/kg/day 11.1 4.8 5.1 .7 .3 .0 44 46 7 3 0 7.1 2.0 4.9 .2 .0 .0 28 69 3 0 0 13.2 5.6 6.3 .8 .4 .0 43 48 6 3 0 10.5 4.1 5.4 .6 .2 .0 38 54 5 2 0 3.10 1.89 .76 .32 .21 .00 9.0 12.7 2.1 1.7 .0 3 3 3 3 3 3 3 3 3 3 3 114 Appendix 4 Individual Clinical Hematology Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ Group: 1 I05353 I05417 Group: 2 I05231 I05352 I05419 Group: 3 I05235 I05351 I05416 ANISO POLY -------- -------- Dose Level: 0 - - - - Dose Level: 2 - - - - - - Dose Level: 20 - - - - - - POIK HYPO TOXNEUT -------- -------- -------- Dosage Unit: mg/kg/day - - - - - - Dosage Unit: mg/kg/day - - - - - - - - - Dosage Unit: mg/kg/day - - - - - - - - - Covance 6329-230 3M T-6889.2 115 Appendix 4 Individual Clinical Hematology Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N RBC X106/L HGB G/DL -------- -------- HCT % -------- MCV FL -------- MCH PG -------- MCHC % -------- PLT X103/L RETIC % -------- -------- RETIC X103/L -------- Dose Level: 0 Dosage Unit: mg/kg/day 4.93 12.5 39.2 79.6 25.5 32.0 414 .4 20 6.99 12.3 42.4 60.6 17.7 29.1 508 .1 7 5.96 12.4 40.8 70.1 21.6 30.5 461 .2 1.457 .14 2.26 13.44 5.52 2.05 66.5 .21 2 2 2 2 2 2 2 2 14 9.2 2 Dose Level: 2 Dosage Unit: mg/kg/day 7.03 13.2 43.6 62.0 18.7 30.3 412 .1 7 5.81 14.9 46.0 79.1 25.6 32.4 404 .4 23 7.38 12.6 43.2 58.5 17.1 29.2 406 .0 0 6.74 13.6 44.3 66.5 20.5 30.6 407 .2 .824 1.19 1.51 11.02 4.52 1.63 4.2 .21 3 3 3 3 3 3 3 3 10 11.8 3 Dose Level: 20 Dosage Unit: mg/kg/day 6.95 12.0 42.0 60.5 17.3 28.6 489 .3 21 5.35 14.2 43.1 80.6 26.6 33.0 605 .4 21 6.41 12.8 40.5 63.2 20.0 31.6 373 .0 0 6.24 13.0 41.9 68.1 21.3 31.1 489 .2 .814 1.11 1.31 10.91 4.78 2.25 116.0 .21 3 3 3 3 3 3 3 3 14 12.1 3 Covance 6329-230 3M T-6889.2 116 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Covance 6329-230 3M T-6889.2 Appendix 4 Individual Clinical Hematology Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS WBC X103/L -------- N-SEG X103/L -------- LYMPH X103/L -------- MONO X103/L -------- EOSIN X103/L -------- BASO X103/L -------- N-SEG% -------- LYMPH% -------- MONO% -------- EOSIN% -------- BASO% -------- Dose Level: 0 Dosage Unit: mg/kg/day 9.9 1.1 7.8 .7 .2 .1 11 79 7 2 1 9.8 2.0 6.7 .4 .7 .0 21 68 4 7 0 9.8 1.6 7.2 .6 .4 .0 16 74 6 4 0 .07 .64 .78 .21 .35 .07 7.1 7.8 2.1 3.5 .7 2 2 2 2 2 2 2 2 2 2 2 Dose Level: 2 Dosage Unit: mg/kg/day 6.2 1.9 3.8 .5 .0 .0 30 61 8 1 0 7.3 1.6 5.4 .2 .0 .0 22 74 3 1 0 10.6 3.1 7.0 .3 .1 .0 30 66 3 1 0 8.0 2.2 5.4 .3 .0 .0 27 67 5 1 0 2.29 .79 1.60 .15 .06 .00 4.6 6.6 2.9 .0 .0 3 3 3 3 3 3 3 3 3 3 3 Dose Level: 20 Dosage Unit: mg/kg/day 9.2 8.2 7.5 4.1 1.9 3.3 4.3 5.8 3.2 .6 .5 .7 .2 .0 .2 .0 .0 .0 45 23 45 47 70 43 7 6 9 2 1 2 0 1 1 8.3 3.1 4.4 .6 .1 .0 38 53 7 2 1 .85 1.11 1.31 .10 .12 .00 12.7 14.6 1.5 .6 .6 3 3 3 3 3 3 3 3 3 3 3 117 Appendix 4 Individual Clinical Hematology Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ ANISO -------- POLY -------- POIK -------- HYPO -------- TOXNEUT -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 - - - - - I05417 - - - - - Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 - - - - - I05352 - - - - - I05419 - - - - - Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 - - - - - I05351 - - - - - I05416 a - - - - - a Plasmodium was observed. Covance 6329-230 3M T-6889.2 118 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Covance 6329-230 3M T-6889.2 Individual Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS GLU MG/DL -------- UN MG/DL -------- CREAT MG/DL -------- T PRO G/DL -------- ALB G/DL -------- GLOB G/DL -------- T BILI MG/DL -------- SBA UMOL/L -------- CHOL MG/DL -------- TRIG MG/DL -------- Dose Level: 0 Dosage Unit: mg/kg/day 59 28 .9 7.0 4.4 2.6 .3 5 152 29 88 17 1.2 10.4 6.0 4.4 .5 13 139 40 74 22 1.0 8.7 5.2 3.5 .4 20.5 7.8 .21 2.40 1.13 1.27 .14 2 2 2 2 2 2 2 9 146 5.7 9.2 2 2 34 7.8 2 Dose Level: 2 Dosage Unit: mg/kg/day 60 16 .9 8.3 5.1 3.2 .2 11 157 40 85 18 1.0 7.1 4.8 2.3 .4 6 170 28 84 20 1.4 9.7 5.4 4.3 .2 9 196 39 76 18 1.1 8.4 5.1 3.3 .3 14.2 2.0 .26 1.30 .30 1.00 .12 3 3 3 3 3 3 3 9 174 2.5 19.9 3 3 36 6.7 3 Dose Level: 20 Dosage Unit: mg/kg/day 63 24 1.4 8.3 4.6 3.7 .2 12 126 39 70 20 .9 7.4 5.0 2.4 .1 11 149 27 60 24 1.2 8.4 4.7 3.7 .2 12 147 28 64 23 1.2 8.0 4.8 3.3 .2 12 141 31 5.1 2.3 .25 .55 .21 .75 .06 .6 12.7 6.7 3 3 3 3 3 3 3 3 3 3 119 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Individual Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS GGT IU/L IU/L IU/L IU/L -------- -------- -------- -------- SDH IU/L -------- CK IU/L -------- AMYLASE LIPASE IU/L IU/L -------- -------- P AMYL U/L -------- Dose Level: 0 Dosage Unit: mg/kg/day 40 38 1144 130 42 52 550 114 2 520 442 2 175 541 42 228 44 275 41 1.4 2 45 9.9 2 847 420.0 2 122 11.3 2 2 348 492 .0 244.0 70.0 2 2 2 43 1.4 2 252 33.2 2 Dose Level: 2 Dosage Unit: mg/kg/day 43 33 1122 209 10 307 415 51 223 65 77 910 122 2 550 427 21 229 37 50 1007 145 3 250 723 67 298 48 14.7 3 53 22.2 3 1013 106.1 3 159 45.1 3 5 369 522 4.4 159.3 174.5 3 3 3 46 23.4 3 250 41.7 3 Dose Level: 20 Dosage Unit: mg/kg/day 45 89 985 116 58 38 986 115 54 50 872 177 4 210 461 4 225 524 6 149 502 44 207 51 280 34 230 52 6.7 3 59 26.7 3 948 65.5 3 136 35.5 3 5 195 496 1.2 40.3 32.0 3 3 3 43 8.5 3 239 37.3 3 Covance 6329-230 3M T-6889.2 120 Appendix 4 Individual Clinical Chemistry Data Males Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ CA MG/DL -------- I PHOS MG/DL -------- NA MMOL/L -------- K MMOL/L -------- CL MMOL/L -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 I05417 10.2 7.9 150 6.1 111 12.6 6.5 166 7.1 113 MEAN S.D. N 11.4 1.70 2 7.2 .99 2 158 11.3 2 6.6 .71 2 112 1.4 2 Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 I05352 I05419 11.3 8.7 159 5.6 109 10.6 7.4 153 6.0 107 11.0 6.5 162 5.3 109 MEAN S.D. N 11.0 .35 3 7.5 1.11 3 158 4.6 3 5.6 .35 3 108 1.2 3 Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 I05351 I05416 11.3 7.2 156 6.1 108 11.1 7.2 154 6.5 109 10.3 6.7 153 5.7 108 MEAN S.D. N 10.9 .53 3 7.0 .29 3 154 1.5 3 6.1 .40 3 108 .6 3 Covance 6329-230 3M T-6889.2 121 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Covance 6329-230 3M T-6889.2 Appendix 4 Individual Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS GLU MG/DL -------- UN MG/DL -------- CREAT MG/DL -------- T PRO G/DL -------- ALB G/DL -------- GLOB G/DL -------- T BILI MG/DL -------- SBA UMOL/L -------- CHOL MG/DL -------- TRIG MG/DL -------- Dose Level: 0 Dosage Unit: mg/kg/day 55 24 1.1 7.2 4.4 2.8 .6 113 11 1.3 9.6 5.5 4.1 .7 6 156 28 3 119 29 84 18 1.2 8.4 5.0 3.4 .6 41.0 9.2 .14 1.70 .78 .92 .07 2 2 2 2 2 2 2 4 138 2.1 26.2 2 2 28 .7 2 Dose Level: 2 Dosage Unit: mg/kg/day 66 14 1.0 8.1 4.9 3.2 .1 60 15 .9 7.1 4.8 2.3 .4 73 16 1.3 8.8 4.7 4.1 .4 1 141 31 5 160 32 9 164 38 66 15 1.1 8.0 4.8 3.2 .3 6.5 1.0 .21 .85 .10 .90 .17 3 3 3 3 3 3 3 5 155 4.0 12.3 3 3 34 3.8 3 Dose Level: 20 Dosage Unit: mg/kg/day 78 14 1.1 8.2 4.4 3.8 .1 51 24 .9 7.4 4.9 2.5 .7 73 21 1.2 8.5 4.7 3.8 .3 1 128 30 2 155 51 0 140 35 67 20 1.1 8.0 4.7 3.4 .4 14.4 5.1 .15 .57 .25 .75 .31 3 3 3 3 3 3 3 1 141 1.0 13.5 3 3 39 11.0 3 122 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Individual Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS AST/SGOT IU/L -------- ALT/SGPT IU/L -------- ALK PHOS IU/L -------- GGT IU/L -------- SDH IU/L -------- CK IU/L -------- AMYLASE IU/L -------- LIPASE IU/L -------- P AMYL U/L -------- Dose Level: 0 Dosage Unit: mg/kg/day 42 34 1066 117 35 52 509 108 3 833 441 3 156 549 42 233 38 270 38 4.9 2 43 12.7 2 788 393.9 2 112 6.4 2 3 494 495 .0 478.7 76.4 2 2 2 40 2.8 2 252 26.2 2 Dose Level: 2 Dosage Unit: mg/kg/day 31 35 1026 182 54 65 872 108 31 46 893 120 4 141 395 3 542 446 2 155 720 64 220 46 244 58 293 39 13.3 3 49 15.2 3 930 83.5 3 137 39.7 3 3 279 520 1.0 227.6 174.8 3 3 3 56 9.2 3 252 37.2 3 Dose Level: 20 Dosage Unit: mg/kg/day 37 66 945 106 54 37 906 92 54 51 837 164 2 181 451 3 194 467 4 138 481 57 213 71 249 44 224 48 9.8 3 51 14.5 3 896 54.7 3 121 38.2 3 3 171 466 1.0 29.3 15.0 3 3 3 57 13.5 3 229 18.4 3 Covance 6329-230 3M T-6889.2 123 Appendix 4 Individual Clinical Chemistry Data Males Day -5 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ CA MG/DL -------- I PHOS MG/DL -------- NA MMOL/L -------- K MMOL/L -------- CL MMOL/L -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 I05417 10.0 7.7 149 4.8 107 11.8 5.9 162 6.1 110 MEAN S.D. N 10.9 1.27 2 6.8 1.27 2 156 9.2 2 5.4 .92 2 108 2.1 2 Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 I05352 I05419 11.1 6.9 159 5.6 113 10.3 7.1 153 5.6 109 9.9 5.2 153 4.3 108 MEAN S.D. N 10.4 .61 3 6.4 1.04 3 155 3.5 3 5.2 .75 3 110 2.6 3 Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 I05351 I05416 10.8 6.1 154 5.3 108 10.5 6.4 153 5.3 108 9.8 6.1 149 4.3 105 MEAN S.D. N 10.4 .51 3 6.2 .17 3 152 2.6 3 5.0 .58 3 107 1.7 3 Covance 6329-230 3M T-6889.2 124 Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N GLU MG/DL -------- UN MG/DL -------- CREAT MG/DL -------- T PRO G/DL -------- ALB G/DL -------- GLOB G/DL -------- T BILI MG/DL -------- SBA UMOL/L -------- CHOL MG/DL -------- TRIG MG/DL -------- Dose Level: 0 Dosage Unit: mg/kg/day 73 26 .8 7.5 4.5 3.0 .3 13 157 23 79 16 1.1 10.6 5.8 4.8 .6 6 146 34 76 21 1.0 9.0 5.2 3.9 .4 10 152 28 4.2 7.1 .21 2.19 .92 1.27 .21 4.9 7.8 7.8 2 2 2 2 2 2 2 2 2 2 Dose Level: 2 Dosage Unit: mg/kg/day 64 17 .8 8.4 4.9 3.5 .4 6 143 49 70 26 .9 7.7 5.0 2.7 .3 9 183 45 62 18 1.2 9.0 4.8 4.2 .3 12 180 61 65 20 1.0 8.4 4.9 3.5 .3 4.2 4.9 .21 .65 .10 .75 .06 3 3 3 3 3 3 3 9 169 3.0 22.3 3 3 52 8.3 3 Dose Level: 20 Dosage Unit: mg/kg/day 68 24 1.1 8.4 4.6 3.8 .1 11 121 35 70 22 .9 7.7 5.0 2.7 .2 12 136 57 66 23 .9 8.7 4.7 4.0 .1 19 140 43 68 23 1.0 8.3 4.8 3.5 .1 14 132 45 2.0 1.0 .12 .51 .21 .70 .06 4.4 10.0 11.1 3 3 3 3 3 3 3 3 3 3 125 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS AST/SGOT IU/L -------- ALT/SGPT IU/L -------- ALK PHOS IU/L -------- GGT IU/L -------- SDH IU/L -------- CK IU/L -------- AMYLASE IU/L -------- LIPASE IU/L -------- P AMYL U/L -------- Dose Level: 0 Dosage Unit: mg/kg/day 45 35 1123 129 49 71 620 121 1 181 435 1 269 519 28 228 30 270 47 2.8 2 53 25.5 2 872 355.7 2 125 5.7 2 1 225 477 .0 62.2 59.4 2 2 2 29 1.4 2 249 29.7 2 Dose Level: 2 Dosage Unit: mg/kg/day 33 31 1151 200 91 82 927 119 36 42 1009 126 1 113 366 4 2149 438 2 121 644 45 204 36 239 51 289 53 32.7 3 52 26.8 3 1029 113.3 3 148 44.9 3 2 794 1.5 1173.2 3 3 483 144.3 3 44 7.5 3 244 42.7 3 Dose Level: 20 Dosage Unit: mg/kg/day 49 80 1062 110 58 36 827 98 57 51 875 171 2 175 433 0 203 503 1 116 502 41 205 39 269 19 227 55 4.9 3 56 22.4 3 921 124.2 3 126 39.1 3 1 165 479 1.0 44.4 40.1 3 3 3 33 12.2 3 234 32.5 3 Covance 6329-230 3M T-6889.2 126 Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ CA MG/DL -------- I PHOS MG/DL -------- NA MMOL/L -------- K MMOL/L -------- CL MMOL/L -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 I05417 10.4 9.0 153 5.5 106 12.7 8.6 170 6.9 113 MEAN S.D. N 11.6 1.63 2 8.8 .28 2 162 12.0 2 6.2 .99 2 110 4.9 2 Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 I05352 I05419 11.0 8.1 158 5.5 113 10.7 9.3 155 5.6 106 9.9 6.7 153 4.8 107 MEAN S.D. N 10.5 .57 3 8.0 1.30 3 155 2.5 3 5.3 .44 3 109 3.8 3 Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 I05351 I05416 11.0 7.5 159 5.7 110 11.4 8.3 157 6.1 107 9.8 6.5 151 4.8 109 MEAN S.D. N 10.7 .83 3 7.4 .90 3 156 4.2 3 5.5 .67 3 109 1.5 3 Covance 6329-230 3M T-6889.2 127 Appendix 4 Individual Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N GLU MG/DL -------- UN MG/DL -------- CREAT MG/DL -------- T PRO G/DL -------- ALB G/DL -------- GLOB G/DL -------- T BILI MG/DL -------- SBA UMOL/L -------- CHOL MG/DL -------- TRIG MG/DL -------- Dose Level: 0 Dosage Unit: mg/kg/day 77 23 .9 7.3 4.6 2.7 .7 90 15 1.0 9.8 5.6 4.2 .3 6 172 33 1 127 32 84 19 1.0 8.6 5.1 3.4 .5 9.2 5.7 .07 1.77 .71 1.06 .28 2 2 2 2 2 2 2 4 150 3.5 31.8 2 2 32 .7 2 Dose Level: 2 Dosage Unit: mg/kg/day 75 15 1.1 8.4 5.1 3.3 .2 80 16 1.2 7.7 5.1 2.6 .1 74 19 1.1 8.9 4.8 4.1 .1 2 137 45 4 149 39 5 155 44 76 17 1.1 8.3 5.0 3.3 .1 3.2 2.1 .06 .60 .17 .75 .06 3 3 3 3 3 3 3 4 147 1.5 9.2 3 3 43 3.2 3 Dose Level: 20 Dosage Unit: mg/kg/day 64 16 1.5 8.0 4.4 3.6 .1 10 107 50 63 22 1.2 7.3 4.8 2.5 .0 1 138 63 67 22 .8 8.2 4.4 3.8 .3 6 170 42 65 20 1.2 7.8 4.5 3.3 .1 2.1 3.5 .35 .47 .23 .70 .15 3 3 3 3 3 3 3 6 138 4.5 31.5 3 3 52 10.6 3 128 Appendix 4 Individual Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N AST/SGOT ALT/SGPT ALK PHOS GGT IU/L IU/L IU/L IU/L -------- -------- -------- -------- SDH IU/L -------- CK IU/L -------- AMYLASE LIPASE IU/L IU/L -------- -------- P AMYL U/L -------- PCOAO IU/G -------- Dose Level: 0 Dosage Unit: mg/kg/day 52 42 1154 118 1 827 451 53 238 2 51 76 680 102 3 195 491 30 247 1 52 59 917 110 2 511 471 42 242 2 .7 24.0 335.2 11.3 1.4 446.9 28.3 16.3 6.4 .7 2 2 2 2 2 2 2 2 2 2 Dose Level: 2 Dosage Unit: mg/kg/day 35 53 1155 177 1 145 344 36 186 0 72 113 1009 115 2 570 501 32 273 0 42 58 1170 115 3 256 654 47 272 3 50 75 1111 136 2 324 500 38 244 1 19.7 33.3 88.9 35.8 1.0 220.4 155.0 7.8 49.9 1.7 3 3 3 3 3 3 3 3 3 3 Dose Level: 20 Dosage Unit: mg/kg/day 43 140 799 80 3 206 382 47 175 5 66 60 626 83 2 216 521 45 277 3 55 48 686 139 0 198 420 26 202 9 55 83 704 101 2 207 441 39 218 6 11.5 50.0 87.8 33.2 1.5 9.0 71.8 11.6 52.8 3.1 3 3 3 3 3 3 3 3 3 3 129 Appendix 4 Individual Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ------ CA MG/DL -------- I PHOS MG/DL -------- NA MMOL/L -------- K MMOL/L -------- CL MMOL/L -------- Group: 1 Dose Level: 0 Dosage Unit: mg/kg/day I05353 I05417 10.2 8.4 149 5.0 104 11.2 5.9 157 5.7 108 MEAN S.D. N 10.7 .71 2 7.2 1.77 2 153 5.7 2 5.4 .49 2 106 2.8 2 Group: 2 Dose Level: 2 Dosage Unit: mg/kg/day I05231 I05352 I05419 11.2 7.1 157 5.2 112 11.0 7.5 156 6.1 107 10.2 5.5 153 4.3 107 MEAN S.D. N 10.8 .53 3 6.7 1.06 3 155 2.1 3 5.2 .90 3 109 2.9 3 Group: 3 Dose Level: 20 Dosage Unit: mg/kg/day I05235 I05351 I05416 10.3 5.8 155 5.1 111 10.9 5.6 152 5.9 107 9.5 5.2 145 4.6 108 MEAN S.D. N 10.2 .70 3 5.5 .31 3 151 5.1 3 5.2 .66 3 109 2.1 3 Covance 6329-230 3M T-6889.2 130 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 131 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 1 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05353 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2500.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 11:10 PROSECTOR: KEVIN BILLINGS RECORDER: JILL PAUS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- KIDNEY (KD) : KIDNEY (KD) : -LIGHT FOCUS(I)/AREA(S); RIGHT ON CORTICAL SURFACE, NEAR >NOT REQUIRED TO BE EXAMINED FOR ANIMAL HILUS: SINGLE TAN AREA, 0.2 CM IN DIAMETER; COLLECTED ON ROUTINE SECTION GENERAL COMMENT (GC) : -EYES - DAVIDSONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), STOMACH, GL (ST), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 132 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 2 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05417 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 3325.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 11:15 PROSECTOR: JERALD W. JOHNSON RECORDER: JAMES T. CALLEN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- STOMACH, GL (ST) : STOMACH, GL (ST) : -RED FOCUS(I)/AREA(S); MUCOSA: MULTIPLE RED AREAS, UP TO 0.8 >NOT REQUIRED TO BE EXAMINED FOR ANIMAL X 0.5 CM; COLLECTED ON ROUTINE SECTION GENERAL COMMENT (GC) : -EYES - DAVIDSONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 133 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 3 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05231 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2795.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 12:43 PROSECTOR: JERALD W. JOHNSON RECORDER: JAMES T. CALLEN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- ADRENAL (AD) : -MINERALIZATION,-MINIMAL, MULTI-FOCAL GENERAL COMMENT (GC) : -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), STOMACH, GL (ST), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE) ------------------------------------------------------------------------------------------------------------------------------ 134 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 4 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05352 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 1980.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 12:40 PROSECTOR: KEVIN BILLINGS RECORDER: JILL PAUS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- TESTIS (TE) : -IMMATURE,-MODERATE, DIFFUSE GENERAL COMMENT (GC) : -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), STOMACH, GL (ST), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 135 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 5 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05419 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 3435.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 11:57 PROSECTOR: JERALD W. JOHNSON RECORDER: JAMES T. CALLEN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- GENERAL COMMENT (GC) : -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS GENERAL INFORMATION (XX) : >NOTE:>LEFT HIND LIMB, NEAR STIFLE JOINT: ANIMAL HAS SIX SUTURES; POSSIBLY CONSISTENT WITH CLINICAL OBSERVATIONS OF SCABS (HIND LEFT) ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), STOMACH, GL (ST), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 136 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 6 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05235 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2635.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 13:09 PROSECTOR: JERALD W. JOHNSON RECORDER: JAMES T. CALLEN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- STOMACH, GL (ST) : STOMACH, GL (ST) : -RED FOCUS(I)/AREA(S); MUCOSA: MULTIPLE RED AREAS, UP TO 0.6 >NOT REQUIRED TO BE EXAMINED FOR ANIMAL X 0.4 CM; COLLECTED ON ROUTINE SECTION GENERAL COMMENT (GC) : -EYES - DAVIDSONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 137 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 7 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05351 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2185.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 11:56 PROSECTOR: KEVIN BILLINGS RECORDER: JILL PAUS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- TESTIS (TE) : -IMMATURE,-MODERATE, DIFFUSE GENERAL COMMENT (GC) : -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), LUNG (LU), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), STOMACH, GL (ST), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 138 APPENDIX 5 Individual Animal Pathology Data Covance 6329-230 3M T-6889.2 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS PAGE: 8 ------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: I05416 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 07/16/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 3120.0 GRAMS DATE AND TIME OF NECROPSY: 07/16/98 13:05 PROSECTOR: KEVIN BILLINGS RECORDER: JILL PAUS POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. JAMES L. CARTER WEIGHER: NOT AVAILABLE ------------------------------------------------------------------------------------------------------------------------------ OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- --------------------------------------------------------- LUNG (LU) : LUNG (LU) : -ADHESION(S); ALL LOBES TO EACH OTHER, PLEURA AND CAUDAL >NOT REQUIRED TO BE EXAMINED FOR ANIMAL LOBES TO DIAPHRAGM: MULTIPLE TAN GRAY FIBROUS ADHESIONS; COLLECTED INTACT STOMACH, GL (ST) : STOMACH, GL (ST) : -RED FOCUS(I)/AREA(S); MUCOSA: SINGLE AREA, 0.5 X 0.2 CM >NOT REQUIRED TO BE EXAMINED FOR ANIMAL GENERAL COMMENT (GC) : -EYES - DAVIDSONS ------------------------------------------ THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), BONE, FEMUR (FE), MARROW, FEMUR (FM), MARROW, STERNUM (SE), BONE, STERNUM (SB), EYE (EY), BRAIN (BR), KIDNEY (KD), HEART (HT), TRACHEA (TR), ESOPHAGUS (ES), THYROID (TY), PARATHYROID (PT), THYMUS (TH), LN, MESENTERIC (MS), AORTA (AO), PITUITARY (PI), SALIV GL, MANDIB (SG), MUSCLE, SKELETAL (SM), SPINAL CORD (SC), NERVE, SCIATIC (SN), DUODENUM (DU), GALLBLADDER (GB), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), SKIN (SK), MAMMARY, MALE (MM), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: LIVER (LI), SPLEEN (SP), PANCREAS (PA), TESTIS (TE), ADRENAL (AD) ------------------------------------------------------------------------------------------------------------------------------ 139 APPENDIX 6 Covance 6329-230 3M T-6889.2 Quality Assurance Statement Summary and Individual Hormone Analyses Data Note: The data included in this appendix were supplied by Ani Lytics Inc. and have been reviewed by the Quality Assurance Unit of Ani Lytics Inc. 140 Covance 6329-230 3M T-6889.2 141 Appendix 6 Summary and Individual Hormone Analyses Data Day -8 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N ESTRONE pg/mL ------- ESTRADIOL pg/mL --------- Dose Level: 0 163.53 236.08 12.39 39.04 199.81 51.301 2 25.72 18.844 2 Dose Level: 2 195.55 187.55 191.95 23.04 30.44 30.63 191.68 4.007 3 28.04 4.328 3 Dose Level: 20 183.99 186.13 213.49 9.10 14.61 12.69 194.54 16.449 3 12.13 2.797 3 ESTRIOL ng/mL ------- THYROID STIMULATING IU/mL ----------- TOTAL TRIIODOTHYRONINE ng/dL ---------------- Dosage Unit: mg/kg/day 0.00 0.00 1.40 1.61 205.95 159.89 0.00 0.000 2 1.51 0.148 2 182.92 32.569 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 1.17 1.18 1.64 187.06 174.59 166.70 0.00 0.000 3 1.33 0.269 3 176.12 10.265 3 Dosage Unit: mg/kg/day 0.00 0.00 0.00 1.50 0.74 1.10 145.43 176.53 172.10 0.00 0.000 3 1.11 0.380 3 164.69 16.823 3 TOTAL THYROXIN g/dL -------- 5.11 4.28 4.70 0.587 2 3.41 5.71 3.12 4.08 1.419 3 3.73 5.99 3.69 4.47 1.317 3 FREE TRIIODOTHYRONINE pg/mL ---------------- 5.08 4.77 4.93 0.219 2 7.61 2.96 5.58 5.38 2.331 3 5.92 3.75 3.44 4.37 1.351 3 FREE THYROXIN ng/dL -------- 1.08 1.14 1.11 0.042 2 1.21 1.17 0.97 1.12 0.129 3 1.41 2.06 0.67 1.38 0.695 3 142 Appendix 6 Summary and Individual Hormone Analyses Data Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH AMMONIUM PERFLUOROOCTANOATE (APFO) IN CYNOMOLGUS MONKEYS Covance 6329-230 3M T-6889.2 ANIMAL NUMBER ------ Group: 1 I05353 I05417 MEAN S.D. N Group: 2 I05231 I05352 I05419 MEAN S.D. N Group: 3 I05235 I05351 I05416 MEAN S.D. N ESTRONE pg/mL ------- ESTRADIOL pg/mL --------- Dose Level: 0 182.68 48.21 13.13 34.20 115.45 95.085 2 23.67 14.899 2 Dose Level: 2 29.31 30.04 21.10 25.88 16.03 17.89 26.82 4.964 3 19.93 5.233 3 Dose Level: 20 14.55 28.80 26.67 10.34 11.50 11.45 23.34 7.687 3 11.10 0.656 3 ESTRIOL ng/mL ------- THYROID STIMULATING IU/mL ----------- TOTAL TRIIODOTHYRONINE ng/dL ---------------- Dosage Unit: mg/kg/day 0.00 0.00 1.68 1.55 188.20 137.22 0.00 0.000 2 1.62 0.092 2 162.71 36.048 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 1.28 1.17 0.98 173.12 150.56 193.34 0.00 0.000 3 1.14 0.152 3 172.34 21.401 3 Dosage Unit: mg/kg/day 0.00 0.00 0.00 1.71 1.06 1.06 138.52 143.31 129.89 0.00 0.000 3 1.28 0.375 3 137.24 6.801 3 TOTAL THYROXIN g/dL -------- 5.50 4.31 4.91 0.841 2 3.84 3.60 5.44 4.29 1.000 3 4.71 5.00 2.62 4.11 1.298 3 FREE TRIIODOTHYRONINE pg/mL ---------------- 4.93 4.72 4.83 0.148 2 5.91 4.98 6.82 5.90 0.920 3 4.55 4.67 3.78 4.33 0.483 3 FREE THYROXIN ng/dL -------- 1.66 1.43 1.55 0.163 2 0.99 1.14 1.69 1.27 0.369 3 1.12 1.19 0.68 1.00 0.276 3 143 APPENDIX 7 Cell Proliferation Report Covance 6329-230 3M T-6889.2 Note: This appendix of the report contains information supplied by Pathology Associates, A Charles River Company and has been reviewed by the Quality Assurance Unit of Pathology Associates, A Charles River Company. 144 Covance 6329-230 3M T-6889.2 145 Covance 6329-230 3M T-6889.2 146 Covance 6329-230 3M T-6889.2 147 Covance 6329-230 3M T-6889.2 148 Covance 6329-230 3M T-6889.2 149 Covance 6329-230 3M T-6889.2 150 Covance 6329-230 3M T-6889.2 151 Covance 6329-230 3M T-6889.2 152 Covance 6329-230 3M T-6889.2 153 Covance 6329-230 3M T-6889.2 154 Covance 6329-230 3M T-6889.2 155 Covance 6329-230 3M T-6889.2 156 Covance 6329-230 3M T-6889.2 157 Covance 6329-230 3M T-6889.2 158 Covance 6329-230 3M T-6889.2 159