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CXR Biosciences Ltd. James Lindsay Place DUNDEE DD1 5JJ REPORT 4-WEEK STUDY TO INVESTIGATE PERFLUOROOCTANE SULFONATE (PFOS)-INDUCED HEPATOMEGALY IN MALE SPRAGUE DAWLEY RATS. CXR STUDY NUMBER: Sponsor: Sponsors Representative: Study Director: CXR0481s 3M Dr John Butenhoff 3M Medical Department 3M Center Building 220-06-W-08, St. Paul, Minnesota 55144-1000 B M Elcombc, CXR Biosciences Test facility "In Life" Start Date: "In Life" Finish Date: CIRCULATION CXR Biosciences, James Lindsay Place, Dundee Technopole, Dundee, DD1 5JJ. and Biological Sciences Resource Unit (BSRU), Dundee University, Dundee, DD1 5EH. 7lhNovember 2006 5lhDecember 2006 Signed Original: Copies to: Study File/Archives Sponsor Study Director Sponsor Author_ Barbara Date CXR0481s Final Report Page 1 of 153 CONTENTS CONTENTS................................................................................................................................ 2 SUMMARY................................................................................................................................ 4 1. INTRODUCTION..................................................................................................................6 2. ANIMALS (Scientific Procedures) ACT 1986 COMPLIANCE..................................... 7 2.1 Licensee Responsibilities.................................................................................................... 7 3. MATERIALS AND M ETHODS.........................................................................................7 3.1 Test Substances..................................................................................................................... 7 3.2 Safety Precautions.................................................................................................................7 3.3 Diet......................................................................................................................................... 7 3.4 A nim als..................................................................................................................................8 3.5 Animal Accommodation and Husbandry.......................................................................... 8 4. EXPERIMENTAL DESIGN.................................................................................................8 5. EXPERIMENTAL PROCEDURES.................................................................................... 9 5.1 Bodyweight...........................................................................................................................9 5.2 Clinical Observations........................................................................................................... 9 5.3 Food Consumption..............................................................................................................10 5.4 Intercurrent D eaths............................................................................................................ 10 5.5 Terminal Procedures.......................................................................................................... 10 6. BIOCHEMICAL MEASUREMENTS..............................................................................11 6.1 Clinical Chemistry..............................................................................................................11 6.2 Peroxisome Proliferation................................................................................................... 11 6.3 DNA Content of L iver.......................................................................................................11 6.4 Total Cytochrome P 4 5 0 .....................................................................................................11 6.5 Lauric Acid 12-Hydroxylation (LAH).............................................................................11 6.6 Pentoxyresorufin-O-depentylation (PROD)....................................................................11 6.7 Testosterone 6P-Hydroxylation........................................................................................ 12 6.8 Western Blotting.................................................................................................................12 6.9 Protein Determination........................................................................................................ 12 6.10 Statistical Analysis........................................................................................................... 12 6.11 Additional Analyses......................................................................................................... 12 7. HISTOPATHOLOGY......................................................................................................... 12 8. SPECIMEN SHIPMENT.....................................................................................................13 9. PROTOCOL AMENDMENTS.......................................................................................... 13 10. RESULTS............................................................................................................................ 13 10.1 Bodyweights, Food Consumption and Liver Weights................................................. 13 10.1.1 Bodyweights...................................................................................................................13 10.1.2 Food consumption......................................................................................................... 14 10.1.3 Liver weights..................................................................................................................16 10.2 Clinical Chemistry........................................................................................................... 18 10.3 Liver Biochemistry..........................................................................................................23 10.3.1 DNA content of liver.................................................................................................... 23 10.3.2 Peroxisome proliferation..............................................................................................25 10.3.3 Total cytochrome P450 content of liver.....................................................................26 10.3.4 Lauric acid hydroxylation........................................................................................... 27 10.3.5 Pentoxyresorufin-O-depentylation..............................................................................28 10.3.6 Testosterone hydroxylation......................................................................................... 29 10.4 Western Blotting.............................................................................................................. 30 10.5 Histopathology..................................................................................................................35 CXR0481s Final Report Page 2 of 153 10.5.1 Liver H & E..................................................................................................................... 35 10.5.2 Hepatic Apoptosis.........................................................................................................35 10.5.3 Cell proliferation in liver..............................................................................................36 10.5.4 Thyroid H & E.................................................................................................................37 10.5.5 Thyroid in situ cell death..............................................................................................37 10.5.6 Cell proliferation in thyroid......................................................................................... 37 11. DISCUSSION..................................................................................................................... 39 12. DATA STORAGE AND ARCHIVING......................................................................... 39 13. REPORT.............................................................................................................................. 40 14. QUALITY ASSURANCE................................................................................................40 15. STUDY DIRECTORS STATEM ENT........................................................................... 40 APPENDIX 1. PATHOLOGY REPORT..............................................................................41 CXR0481s Final Report Page 3 of 153 SUMMARY The mean ingested doses of Test Items calculated for the duration of the study were: 1.66 0.03 (PFOS 20ppm), 7.90 0.17 (PFOS 100ppm), 41.79 0.66 (PB) and 4.65 0.07 (Wy14,643) mg Test Item / kg bwt / day. No adverse clinical observations were made throughout the study. Administration of dietary PFOS (100ppm) and Wy14,643 (50ppm) to rats led to decreased body weights after 28 days of exposure. This effect was not observed in the animals receiving dietary PFOS (20ppm) or PB (500ppm). Absolute liver weights were increased with time following exposure to both dose levels of PFOS, PB and Wy14,643, reaching 116%, 114%, 126% and 161% of control values at Day 29 respectively. Relative liver weights were markedly increased by all compounds studied. Increases to approximately 116%, 145%, 121% and 180% of control values were observed following 28 days of exposure. A lack of overt hepatotoxicity was indicated for all compounds by the absence of toxicologically significant increases in plasma ALT and AST. Plasma cholesterol and triglycerides were markedly decreased by PFOS and Wy14,643. PFOS was especially potent in this respect. CN- - insensitive palmitoyl CoA oxidation was induced by Wy14,643 and both dose levels of PFOS in a time- and dose-dependent manner. PB was without effect. All compounds studied had marked and characteristic effects on cytochromes P450 and related monooxygenase activities. At Day 29, 20ppm PFOS, 100ppm PFOS and PB increased total hepatic microsomal cytochrome P450 to 186%, 333% and 218% of control values. Wy14,643 had no effect on total cytochrome P450 content. However, all compounds altered the P450 isoform profiles substantially. SDS-PAGE and western blotting of hepatic microsomes, using specific antibodies to cytochrome P450 isoforms, revealed the induction of CYP2B, CYP2E1 and CYP3A1 by phenobarbital and PFOS. Wy14,643 had no effect on the expression of these isoforms. Wy14,643 was a rapid and potent inducer of the CYP4A1 isoform with marked effects seen 24hr after the first dose of compound. PFOS also induced CYP4A1, however the kinetics of induction were much slower. PB had no effect on CYP4A1 expression. These compound-induced alterations in cytochrome P450 isoform profiles were CXR0481s Final Report Page 4 of 153 reflected by changes in pentoxyresorufin-O-depentylation (CYP2B), testosterone-6Phydroxylation (CYP3A1) and lauric acid-12-hydroxylation (CYP4A1). In the liver, histopathology revealed decreased glycogen vacuolation by all three compounds. However this was more marked and seen earlier following the administration of Wy14,643. A minimal to slight, mainly centrilobular hepatocellular hypertrophy was noted, dose related in severity, in the animals of the 20ppm PFOS, 100ppm PFOS and 500ppm PB groups after 7 and 28 days of treatment. In the animals treated with 50ppm Wy14,683 a similar but mainly diffuse hepatocellular hypertrophy was seen in all rats at all time points. The severity of this finding was also increased with the duration of treatment 100ppm PFOS, PB and Wy14,643 administration decreased the hepatic apoptotic index at all time points studied. This effect was seen after 1 day and was maintained to Day 29. Apoptotic indices of 40%, 40% and 50% of control values for PFOS, PB and Wy14,643, respectively, were recorded after 28 days of treatment. 20ppm PFOS, 100ppm PFOS, PB and Wy14,643 administration all increased the hepatocellular labelling index (S-phase) at all time points studied. These increases were 2-, 3-, 6- and 10-fold, respectively, 1 day after commencement of treatment. After 7 days of treatment, the levels of S-phase activity reached a maximum of 4-, 5-, 7- and 25-fold, respectively. By 28 days of treatment, the labelling indices were apparently decreasing toward control values and were 2-, 5-, 2- and 11-fold respectively of concurrent control values. In the thyroid gland, no microscopic findings considered to be related to the treatment with the test items were recorded. The number of apoptotic cells found in all treatment groups, at all time points, was minimal. Regardless of treatment, it was usual to find either no cells or just one cell per thyroid staining in the TUNEL assay as apoptotic. Wy14,643 administration increased the thyroid follicular cell labelling index (Sphase) after 1 and 7 days of treatment, whilst PB administration increased the level of S-phase activity only after 7 days of treatment. After 28 days of treatment the labelling indices had returned to control values in both Wy14,643- and PB- treated animals. 20ppm PFOS and 100ppm PFOS treatment had no effect on S-phase at any time point studied. CXR0481s Final Report Page 5 of 153 1. INTRODUCTION A variety of non-genotoxic chemicals elicit hepatocellular adenomas and carcinomas when administered to rats in long-term studies. The appearance of tumours is preceded by the rapid and early stimulation of hepatomegaly. The liver enlargement seen following the administration of such "liver growth carcinogens" to rats is a result of both hypertrophy and hyperplasia. Depending on the particular chemical, the hypertrophy is characterised by proliferation of peroxisomes and/or smooth endoplasmic reticulum, induction of peroxisomal enzymes and induction of cytochrome P450 isoforms. The hyperplasia is characterised by stimulation of hepatocellular proliferation (S-phase, labelling index) and inhibition of apoptosis. PFOS has previously been shown to elicit liver growth in rats, and this study aimed to characterise the hepatomegaly in these terms. Thyroid follicular cell adenoma effects have also been reported in male rats after the administration of 20ppm PFOS in the diet for one year followed by up to one year of control rat diet. However, male rats given PFOS in the diet at the same dose continuously for up to two years did not experience an increase in the tumour. This study used liver, serum, thyroid and duodenal (for assessing BrdU labelling) samples generated over a time course including exposure for 1, 7 and 28 days. Phenobarbital (PB) and Wy14,643 were used as positive controls. The following parameters were studied: Bodyweights Liver weights (relative and absolute) Clinical chemistry (5 parameters). Haematoxylin and eosin (H&E) histopathology (liver and thyroid) Apoptotic indices analysed using the TUNEL assay (liver and thyroid) BrdU incorporation analysed as a measure of cell proliferation (liver and thyroid) The DNA content of liver was measured using the diphenylamine reaction Acyl CoA oxidase activity was determined in liver heavy pellets as CN--insensitive palmitoyl CoA oxidation Total P450 content was determined in liver microsomes Lauric acid hydroxylation (LAH, CYP4A) was determined in liver microsomes Pentoxyresorufin-O-depentylation (PROD, CYP2B) was determined in liver microsomes Testosterone hydroxylation (CYP 3A) was determined in liver microsomes Expression of CYPs 2B1/2, 2E1, 3A1 and 4A1 by SDS-PAGE and Western blotting in liver microsomes. These blots were only performed on controls, high dose PFOS, Wy 14,643 and PB livers CXR0481s Final Report Page 6 of 153 2. ANIMALS (Scientific Procedures) ACT 1986 COMPLIANCE The in-life procedures undertaken during the course of this Study were subject to the provisions of the United Kingdom Animals (Scientific Procedures) Act 1986. The Act, administered by the UK Home Office, regulates all scientific procedures in living animals which may cause pain, suffering, distress or lasting harm and provides for the designation of establishments where procedures may be undertaken, the licensing of trained individuals who perform the practical techniques, and the issue of project licences for specified programmes of work. This Study complied with all applicable sections of the Act and the associated Codes of Practice for the Housing and Care of Animals used in Scientific Procedures and the Humane Killing of Animals under Schedule 1 to the Act, issued under section 21 of the Act. 2.1 Licensee Responsibilities The procedure performed was PD1 as detailed in the PD project licence (60/3005). The severity limit for this procedure is MODERATE. Mr F Simeons returned the animals at the start of the study under the PD project licence. 3. MATERIALS AND METHODS 3.1 Test Substances Perfluorooctane sulfonate potassium salt (PFOS K+, Lot no. 217, purity 87.6%) supplied by 3M Company, St. Paul Minnesota, USA, was a white powder. Wy14,643 was purchased from Alexis Biochemicals (ALX-270-198). Phenobarbital, sodium salt, was purchased from Sigma (P5178). They were handled, and their identity verified, according to CXR Biosciences SOP 0010. This SOP describes routine procedures for the receipt, identification, labelling, handling, sampling and storage of Test and Reference Items. These procedures are implemented to ensure that: Test and Reference Items used are those Items specified in Study Protocols. Handling and storage of Test and Reference Items preserves their identity and integrity during use. 3.2 Safety Precautions The normal safety precautions as detailed in the relevant Sop's and COSHH assessments were applied, no additional precautions were considered necessary. 3.3 Diet RM1 powdered diet (Special Diet Services Ltd., Stepfield, Witham, Essex, UK) was used; Biological Sciences Resource Unit (BSRU) hold the specification of the diet. The Sponsor requested the dietary route of administration. CXR0481s Final Report Page 7 of 153 The test diets, containing 20ppm PFOS, 100ppm PFOS, 500ppm PB or 50ppm Wy14,643 were prepared by CXR Biosciences (Laboratory Method Sheet [LMS] TSC-002) and samples were retained for analysis by the Sponsor. Rats were fed RM1 powdered diet or test diets ad libitum throughout the study. 3.4 Animals A sufficient number of male 6-7 week old Charles River Sprague Dawley (CD) rats were obtained from Charles River, UK. On arrival in the BSRU the rats were housed 2 per cage on sawdust in solid -bottom, polypropylene cages. The rats were acclimatised for a period of at least 5 days before use. No animals were excluded from the study. Environmental enhancing additions were not used in the cages prior to the start of the Study. The Sprague Dawley (CD) rat was the test system of choice for this study because previous work on PFOS has been undertaken using this strain. In addition, CXR Biosciences has extensive experience in the use of this strain of rat, and the strain is well accepted by the regulatory authorities. 3.5 Animal Accommodation and Husbandry The environment was controlled in the animal room to provide conditions suitable for the Sprague Dawley (CD) strain of rat. The temperature was maintained within a range of 1923oC and relative humidity within a range of 40-70%. There were a nominal 14-15 air changes per hour. Twelve-hour periods of light were cycled with twelve-hour periods of darkness. Drinking water was taken from the local supply and provided in bottles. Drinking water was provided ad libitum prior to and throughout the study. Prior to the start of the study the rats were allowed powdered RM1 diet ad libitum. 4. EXPERIMENTAL DESIGN The animals were uniquely numbered by ear-punch and randomly allocated to groups up to one week after arrival. An experimental card was placed on each cage and showed the project licence code, treatment group, study number, sex and individual numbers of the rats within, and identified the Home Office Licensee. In addition, these cards were colour-coded to correlate with the coding for the treatment group on the diet boxes and jars. The study consisted of one control and four test groups, each containing 30 male animals. Control animals received powdered RM1 diet ad libitum for the duration of the study. The test groups of animals were administered either 20ppm PFOS, 100ppm PFOS, 500ppm PB or 50ppm Wy14,643 in the diet ad libitum for either 1, 7 or 28 days. The concentration of PFOS in powdered RM1 diet was prepared with purity correction. The dietary concentrations of PB and Wy14,643 were made up without any correction for purity. The corresponding PFOS, CXR0481s Final Report Page 8 of 153 PB, and Wy14,643 concentrations in the powdered RM1 diet will be analyzed by the Sponsor. Ten animals from each group were sacrificed on Days 2, 8 and 29. Animals were implanted with osmotic pumps (Alzet 2ML1) containing bromodeoxyuridine (BrdU, 15mg/mL in phosphate buffered saline [PBS], pH7.4) 5 days before termination, whilst still receiving the treatment regimen (Table 1). Animals sacrificed after exposure to diet for 1 day were administered BrdU (15mg/mL in PBS, 5mL/kg bodyweight) by subcutaneous injection 2 hours prior to sacrifice. TABLE 1 EXPERIMENTAL DESIGN Group Colour code Animal No 1 Blue 1 - 10 11 - 20 21 - 30 2 Green 31 - 40 41 - 50 51 - 60 3 Yellow 61 - 70 71 - 80 81 - 90 4 Red 91 - 100 101 - 110 111 - 120 5 Black 121 - 130 131 - 140 141 - 150 * = su ^cutaneous injection Treatment Control Arrival 31/10/06 PFOS 20ppm 31/10/06 PFOS 100ppm 31/10/06 PB 500ppm 31/10/06 Wy-14,643 31/10/06 50ppm Dates Start Implant 07/11/06 07/11/06 07/11/06 07/11/06 07/11/06 08/11/06* 09/11/06 30/11/06 08/11/06* 09/11/06 30/11/06 08/11/06* 09/11/06 30/11/06 08/11/06* 09/11/06 30/11/06 08/11/06* 09/11/06 30/11/06 Kill 08/11/06 14/11/06 05/12/06 08/11/06 14/11/06 05/12/06 08/11/06 14/11/06 05/12/06 08/11/06 14/11/06 05/12/06 08/11/06 14/11/06 05/12/06 5. EXPERIMENTAL PROCEDURES 5.1 Bodyweight The bodyweight of each rat was recorded at the start of the study. The animals were weighed weekly on the same day of the week. All animals were weighed prior to termination. Bodyweights were recorded electronically and records of the weights were stored in the Study File. 5.2 Clinical Observations Prior to the start of the study, all rats were observed to ensure that they were physically normal and that they exhibited normal activity. Each rat was observed at least once daily CXR0481s Final Report Page 9 of 153 during the study. Clinical abnormalities of individual animals were recorded in the Study Diary. This was kept in the BSRU until completion of the in life phase of the study and then transferred to the Study File for retention and archiving. 5.3 Food Consumption Food consumption was measured every time the diet jars were changed / refilled, and when the animals were sacrificed. Animals on diet for 7 days or greater had their diet jars weighed on the same day of the week, regardless of refilling, so that weekly food consumption could be recorded. Evidence of excessive diet spillage was noted if it occurred. 5.4 Intercurrent Deaths There were no rats requiring euthanasia during the study, nor were there any intercurrent deaths. 5.5 Terminal Procedures On the day of termination the rats were weighed and transferred to the post mortem room. The rats were killed by exposure to a rising concentration of CO2. Approximately 6 to 10 mL of venous blood was taken by cardiac puncture and dispensed at equal volume into uncoated tubes (to obtain serum) and lithium/heparin-coated tubes (to obtain plasma). The tubes were mixed on a roller for 10 min then cooled on ice. Serum and plasma were prepared by centrifugation (2,000 rpm for 10 min at 8 - 10C) then stored at approximately -70oC pending analysis. The pellet was discarded. The liver was removed from each animal and weighed. 2 samples of liver, approximately 2mm strips, were taken, one from the Left lobe and one from the Median lobe. These were placed in 10% neutral buffered formaldehyde (NBF), for approximately 48hours, for BrdU immunohistochemistry and analysis of the apoptotic index. 2 samples of liver, approximately 2mm strips, were taken, one from the Left lobe and one from the Median lobe. These were placed in NBF, for at least 1 week, for H&E histopathology. 1 sample of liver, of approximately 0.25g, was weighed and then flash frozen in liquid nitrogen then stored at approximately -70oC for DNA determination. 2 samples of liver, of approximately 1g each, were flash frozen in liquid nitrogen then stored at approximately -70oC prior to dispatch, on dry ice, to the Sponsor for further analytical determination under a separate protocol The remainder of the liver was weighed and scissor-minced in ice-cold 1.15% (w/v) KCl prior to subcellular fractionation according to Laboratory Method Sheet (LMS) Cent-001. Samples of homogenate, heavy pellet and microsomes were stored at approximately -70oC prior to analysis. Samples of cytosol were stored at approximately -70oC prior to CXR0481s Final Report Page 10 of 153 dispatch, on dry ice, to the Sponsor for further analytical determination under a separate protocol. 1 sample of duodenal small intestine, of approximately 1cm, was taken and placed in NBF, for approximately 48 hours, as a positive control for BrdU immunohistochemistry. The thyroid was removed from each animal, including the parathyroid gland together with trachea and oesophagus. This tissue was placed in NBF, for approximately 48 hours, for BrdU immunohistochemistry, analysis of the apoptotic index and H&E histology. 6. BIOCHEMICAL MEASUREMENTS 6.1 Clinical Chemistry Plasma samples, prepared as described above, were assayed for ALT, AST, cholesterol, glucose and triglycerides using a Roche Integra 400 automated analyser according to the manufacturer's instructions. System control samples and calibration standards were those supplied by the manufacturer. Results were maintained in the Study File. 6.2 Peroxisome Proliferation CN- -insensitive acyl CoA oxidation was determined spectrophotometrically in liver heavy pellet, using palmitoyl CoA as a substrate, according to LMS Spec003. Results were expressed as nmol NAD+ reduced/min/mg protein. Results were maintained in the Study File. 6.3 DNA Content of Liver The DNA content of liver was measured spectrophotometrically in whole tissue using the diphenylamine reaction, according to LMS Spec005. Results were expressed as pg DNA/g liver and mg DNA/whole liver. Results were maintained in the Study File. 6.4 Total Cytochrome P450 The total cytochrome P450 content of liver microsomes was determined by measuring the carbon monoxide difference spectrum of ferrocytochrome P450 according to LMS Spec002. Results were expressed as nmol total P450/mg protein. Results were maintained in the Study File. 6.5 Lauric Acid 12-Hydroxylation (LAH) The activity of CYP4A in liver microsomes was determined using lauric acid and LC-MSMS. Results were expressed as nmols 12-OH lauric acid formed/10minutes/mg protein. Results were maintained in the Study File. 6.6 Pentoxyresorufin-O-depentylation (PROD) The activity of CYP2B in liver microsomes was determined spectrofluorometrically by the CXR0481s Final Report Page 11 of 153 formation of resorufin from pentoxyresorufin, according to LMS Fluor-002. Results were expressed as pmols resorufin formed/minute/mg protein. Results were maintained in the Study File. 6.7 Testosterone 60-Hydroxylation The activity of CYP3A in liver microsomes was determined by HPLC followed by UV detection, according to LMS HPLC-006. Results were expressed as nmols hydroxytestosterone formed/minute/mg protein. Results were maintained in the Study File. 6.8 Western Blotting Expression of CYPs 2B1/2, 2E1, 3A1 and 4A1 was carried out by SDS-PAGE and Western blotting, using liver microsomes, according to LMS Elec006, Elec005 and Elec003. Results were maintained in the Study File. 6.9 Protein Determination The protein concentration of tissue heavy pellets and microsomes was determined in aqueous solutions using a modification of the method of Lowry et al., (1951) and bovine serum albumin standards, according to LMS Spec001. Results were maintained in the Study File. 6.10 Statistical Analysis Statistical comparisons between treated and control groups have been undertaken for all numerical data sets. The statistical analyses performed are documented with the results. 6.11 Additional Analyses The Sponsor has requested no additional analyses. 7. HISTOPATHOLOGY Following fixation, all fixed liver and thyroid samples were processed, embedded and 5 p,M sections cut according to LMS Pat-005, Pat-006 and Pat-007. Sections were: Stained using H&E, according to LMS Pat-010, Pat-011 and Pat-012. Analysed for in situ cell death. The method used was an indirect TUNEL labelling assay, according to LMS Pat-014. Analysed for BrdU incorporation as a measure of cell proliferation. The method used was an indirect BrdU labelling assay, according to LMS Pat-008. H&E sections were evaluated by Dr. med. vet. Ortwin Vogel, GoethestraBe 26, 24116 Kiel, Schleswig-Holstein, Deutschland. The histopathology report was maintained in the Study File and as an appendix to the Study Report. CXR0481s Final Report Page 12 of 153 8. SPECIMEN SHIPMENT The powdered RM1 diets, cytosol, serum, and the liver samples designated for further analytical determination by the Sponsor were shipped on dry ice and sent to: Dave Ehresman 3M Company 3M Center Building 236-C148 St. Paul, MN 55144 9. PROTOCOL AMENDMENTS There were no changes to the protocol, and no Protocol Amendments have been signed by the Study Director. 10. RESULTS 10.1 Bodyweights, Food Consumption and Liver Weights. 10.1.1 Bodyweights. Administration of dietary PFOS (100ppm) and Wy14,643 (50ppm) to rats lead to decreased body weights (bwt) after 28 days of exposure (Table 2). This effect was not observed in the animals receiving dietary PFOS (20ppm) or PB (500ppm). No adverse clinical observations accompanied the decreased body weights. Table 2. Terminal Bodyweights Days Bodyweight (g) on 0ppm PFOS PFOS PB WY14,643 diet 20ppm 100ppm 500ppm 50ppm 1 261.6 17.3a 249.8 15.0 255.6 20.3 259.7 21.7 257.9 16.0 (100 6.6) (95.5 5.7) (97.7 7.8) (99.3 8.3) (98.6 6.1) 7 300.9 15.6 299.1 15.0 292.3 17.0 307.7 26.4 299.8 10.6 (100 5.2) (99.4 5.0) (97.1 5.7) (102.3 8.8) (99.6 3.5) 28 391.9 26.1 392.7 32.7 309.1 15.1*** 409.8 37.0 350.8 29.4** (100 6.7) (100.2 8.3) (78.9 3.9) (104.6 9.5) (89.5 7.5) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; **p<0.01; *** p<0.001. CXR0481s Final Report Page 13 of 153 500 450 400 350 300 250 200 150 100 50 0 1 Day Terminal Bodyweights Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm 7 Days Days on diet 28 Days 10.1.2 Food consumption. The food consumption of the PFOS 20ppm, PB and Wy14,643-treated rats was essentially the same as control values throughout the experimental period. Administration of PFOS 100ppm decreased food consumption throughout the experimental period (Table 3.). When diet consumption was calculated per unit body weight, that of PFOS 100ppmtreated animals was less than control values for the second and third weeks. The Wy14,643-treated rats consumed more diet per unit body weight during week four of the experiment. CXR0481s Final Report Page 14 of 153 Table 3. Food Consumption Days on diet 7 14 21 28 0ppm 190.0 15.8a (100 8.31) 204.7 21.5 (100 10.5) 198.6 18.6 (100 9.4) 176.9 21.9 (100 12.4) Diet Consumed (g) / Rat / Week PFOS PFOS PB 20ppm 100ppm 500ppm 193.1 5.5 179.9 12.3 196.0 13.7 (101.6 2.9) (94.7 6.5) (103.2 7.2) 193.1 12.8 161.9 7.1*** 205.0 18.0 (94.3 6.2) (79.1 3.5) (100.1 8.8) 188.4 13.3 140.9 9.2*** 195.9 17.0 (94.9 6.7) (70.9 4.6) (98.6 8.6) 174.6 9.5 (98.7 5.4) 126.8 19.1** 187.9 14.4 (71.7 10.8) (106.2 8.1) Wy14,643 50ppm 205.8 5.8* (108.3 3.1) 195.6 23.2 (95.6 11.4) 198.0 24.1 (99.7 12.1) 190.2 21.0 (107.5 11.9) Days on diet 1 7 14 21 28 0ppm 99.9 20.6 (100 20.7) 90.2 5.5 (100 6.1) 86.4 8.3 (100 9.6) 77.9 8.8 (100 11.2) 64.5 7.8 (100 12.0) Diet Consumed (g) / Kg Bwt / Day PFOS 20ppm PFOS 100ppm PB 500ppm 106.1 7.8 110.5 9.9 106.2 4.1 (106.3 7.8) (110.6 9.9) (106.3 4.1) 92.3 3.5 87.9 5.0 91.1 3.0 (102.4 3.9) (97.5 5.6) (101.0 3.4) 80.7 4.1 75.6 3.0* 82.3 3.8 (93.4 4.7) (87.5 3.5) (95.3 4.4) 73.5 5.8 64.8 4.3** 72.8 1.6 (94.3 7.5) (83.1 5.6) (93.5 2.1) 63.5 2.8 58.1 7.1 65.6 2.3 (98.5 4.4) (90.2 11.0) (101.7 3.5) Wy14,643 50ppm 118.4 8.8 (118.6 8.8) 98.1 1.4** (108.8 1.6) 86.6 7.1 (100.3 8.2) 84.7 7.0 (108.8 9.0) 77.4 4.4** (120.0 6.9) Days Test Item Consumption: mg Test Item / Kg Bwt / Day on 0ppm diet PFOS 20ppm PFOS 100ppm PB 500ppm Wy14,643 50ppm 1 00 2.12 0.16 11.05 0.99 53.09 2.05 5.92 0.44 7 00 1.85 0.07 8.79 0.50 45.54 1.52 4.90 0.07 14 0 0 1.61 0.08 7.56 0.30 41.15 1.88 4.33 0.35 21 0 0 1.47 0.12 6.47 0.43 36.39 0.80 4.24 0.35 28 0 0 1.27 0.06 5.62 0.76 32.79 1.13 3.87 0.22 aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. CXR0481s Final Report Page 15 of 153 The mean ingested doses of PFOS 20ppm, PFOS 100ppm, PB and Wy14,643, calculated for the duration of the study, were 1.66 0.03, 7.90 0.17, 41.79 0.66 and 4.65 0.07 mg Test Item / kg bwt / day respectively. 10.1.3 Liver weights. Absolute liver weights were increased with time following exposure to both dose levels of PFOS, PB and Wy14,643, reaching 116%, 114%, 126% and 161% of control values at Day 29, respectively (Table 4). Table 4. Absolute Liver Weights Days Liver weight (g) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 11.81 1.13a 11.86 1.19 11.99 1.13 12.02 1.71 12.43 0.82 (100 9.6) (100.4 10.1) (101.5 9.5) (101.8 14.5) (105.2 6.9) 7 14.66 1.07 14.66 1.47 16.07 1.64* 17.30 2.08** 21.91 1.54*** (100 7.3) (100.0 10.1) (109.6 11.2) (118.0 14.2) (149.5 10.5) 28 16.03 1.61 18.58 1.85** 18.33 1.21** 20.26 2.90*** 25.79 3.85*** (100 10.0) (116.0 11.53) (114.4 7.5) (126.4 18.1) (160.9 24.0) a Values are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Absolute Liver Weights 35 -|-------------------------------------------------------------------------------------------------------------------------------------------------------- 30 25 Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm 1 Day CXR0481s Final Report 7 Days Days on diet Page 16 of 153 28 Days After normalisation with respect to bodyweight, relative liver weights were markedly increased by all compounds studied within 28 days of treatment. Increases to approximately 116%, 145%, 121% and 180% of control values were observed following 28 days of exposure (Table 5). Table 5. Liver/bodyweight ratio (%) Days Liver/bodyweight ratio (%) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 4.51 0.24a 4.74 0.26 4.69 0.22 4.61 0.31 4.82 0.20** (100 5.24) (105.02 5.80) (103.94 4.80) (102.19 6.82) (106.87 4.44) 7 4.88 0.37 4.90 0.38 5.49 0.41** 5.62 0.36*** 7.31 0.44*** (100 7.56) (100.44 7.82) (112.64 8.50) (115.14 7.28) (149.83 8.95) 28 4.09 0.26 4.74 0.37*** 5.93 0.33*** 4.93 0.35*** 7.32 0.55*** (100 6.47) (115.95 9.08) (145.20 8.15) (120.57 8.56) (179.20 13.38) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Relative Liver Weights 9 Control PFOS 20ppm - PFOS 100ppm PB 500ppm W Y-14,643 50ppm 6 5 4 2 0 1 Day 7 Days Days on Diet 28 Days CXR0481s Final Report Page 17 of 153 10.2 Clinical Chemistry The lack of overt hepatotoxicity was indicated for all compounds by the absence of toxicologically significant increases in plasma ALT and AST (Tables 6 and 7). Table 6. Plasma Alanine Aminotransferase Activity Days ALT (U/L) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 142.4 30.1a 119.6 17.0 133.5 39.1 108.8 28.2* 96.2 12.5*** (100 21.1) (84.0 12.0) (93.7 27.4) (76.4 19.8) (67.5 8.8) 7 102.3 16.5 99.1 16.8 87.2 12.6* 87.3 12.5* 88.9 23.6 (100 16.2) (96.8 16.4) (85.2 12.3) (85.3 12.2) (86.9 23.1) 28 80.7 9.9 78.6 15.9 88.9 14.0 79.3 6.0 60.7 17.4** (100 12.2) (97.4 19.7) (110.2 17.4) (98.3 7.5) (75.2 21.6) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Plasma ALT Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm 140 ~ 100 - D I- < 80 60 20 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 18 of 153 Table 7. Plasma Aspartate Transaminase Activity Days AST (U/L) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 160.8 44.7a 141.2 20.4 160.4 50.3 121.0 30.9* 122.6 25.7* (100 27.8) (87.8 12.7) (99.8 31.3) (75.2 19.2) (76.2 16.0) 7 149.9 27.8 158.8 33.4 169.9 28.8 180.3 45.3 146.0 35.1 (100 18.6) (105.9 22.3) (113.4 19.2) (120.3 30.2) (97.4 23.4) 28 100.6 11.3 98.7 16.4 117.1 13.5* 104.5 15.2 97.9 16.0 (100 11.2) (98.2 16.3) (116.5 13.5) (103.9 15.1) (97.3 16.0) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Plasma AST Control PFOS 20ppm - PFOS 100ppm PB 500ppm W Y-14,643 50ppm 50 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 19 of 153 In general, mean plasma cholesterol concentrations were decreased by both Wy14,643 and PFOS at all time points (Table 8), while mean plasma glucose concentrations were initially decreased after 1 day by Wy14,643 but were significantly decreased after 28 days by PFOS at 100ppm (Table 9). Table 8. Plasma Total Cholesterol Concentration Days Total Cholesterol (mmol/L) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 2.39 0.43a 2.12 0.39 1.98 0.24* 2.19 0.46 1.26 0.22*** (100 17.86) (89.01 16.26) (83.14 10.05) (91.95 19.42) (52.75 9.09) 7 2.10 0.30 1.61 0.25** 1.29 0.38*** 2.47 0.77 1.44 0.30*** (100 14.33) (77.00 11.95) (61.74 18.07) (117.65 36.72) (68.70 14.15) 28 2.11 0.46 1.21 0.28*** 0.34 0.14*** 2.13 0.49 1.74 0.46 (100 21.89) (57.60 13.41) (16.09 6.84) (101.00 23.20) ( 82.62 )21.83 aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Plasma Cholesterol 3.5 Control 3.0 PFOS 20ppm PFOS 100ppm PB 500ppm 2.5 W Y -14,643 50ppm 2.0 E 0 1 1.5 o .Oe 1.0 0.5 0.0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 20 of 153 Table 9. Plasma Glucose Concentration Days Glucose (mmol/L) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 19.28 3.51a 17.09 2.52 20.19 1.71 19.63 2.64 16.68 1.64* (100 18.20) (88.61 13.09) (104.72 8.89) (101.79 13.67) (86.50 8.51) 7 16.58 2.14 16.27 2.17 17.98 2.10 17.06 6.28 15.17 1.30 (100 12.93) (98.12 13.12) (108.42 12.64) (102.86 37.89) (91.51 7.82) 28 21.27 3.95 20.63 2.78 15.77 2.59** 19.17 1.79 18.27 2.80 (100 18.56) (97.00 13.08) (74.17 12.19) (90.13 8.43) ( 85.92 )13.17 aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Plasma Glucose 30 Control PFOS 20ppm PFOS 100ppm 25 PB 500ppm W Y-14,643 50ppm 20 15 10 5 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 21 of 153 After 1 day on diet, mean plasma triglyceride concentrations were slightly elevated by PB and reduced by Wy14,643. After 7 days there was a marked reduction produced by the administration of 100ppm PPFOS and Wy14,643. The plasma triglyceride concentrations continued to decrease in the animals administered 100ppm PFOS, but recovered to near control levels in the animals administered Wy14,643 (Table 10). Table 10. Plasma Triglyceride Concentration Days Triglycerides (mmol/L) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 0.95 0.27a 1.15 0.27 1.02 0.24 1.56 0.70* 0.63 0.20** (100 28.4) (121.5 28.5) (108.1 24.9) (164.7 74.2) (66.0 21.5) 7 1.75 0.67 1.32 0.51 0.79 0.29*** 1.73 1.02 0.84 0.24*** (100 38.5) (75.3 29.2) (45.4 16.3) (99.2 58.1) (48.1 14.0) 28 1.82 0.76 1.36 0.58 0.20 0.06*** 1.13 0.71 1.52 0.63 (100 41.7) (75.1 32.1) (10.9 3.4) (62.4 39.0) (83.6 34.7) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. 3.0 2.5 2.0 1.5 1.0 0.5 0.0 1 Day Plasma Triglycerides Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm 7 Days Days on diet 28 Days CXR0481s Final Report Page 22 of 153 10.3 Liver Biochemistry 10.3.1 DNA content of liver The mean hepatic concentration of DNA was initially increased by the 100ppm PFOS, PB and Wy14,643 treatments after 1 day on diet. Continued treatment resulted in decreased within-group mean liver DNA concentrations at all further time-points examined (Table 11). Table 11. Liver DNA Concentration (mg DNA/g liver) Days Liver DNA (mg/g liver) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 1.82 0.12a 1.86 0.22 2.14 0.11*** 2.14 0.11*** 2.24 0.21*** (100 6.7) (102.5 11.9) (117.5 6.2) (117.5 6.3) (123.4 11.8) 7 1.93 0.19 1.71 0.16** 1.93 0.13 1.86 0.32 1.67 0.14** (100 9.6) (88.5 8.3) (99.7 6.8) (96.5 16.6) (86.7 7.1) 28 1.93 0.27 1.74 0.15 1.71 0.14* 1.78 0.15 1.82 0.13 (100 13.8) (90.2 7.6) (88.8 7.5) (92.4 7.8) (94.2 6.9) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. 3.0 2.5 2.0 z 1.5 Q 1.0 0.5 0.0 Liver DNA concentration (mg DNA/g liver) Control PFOS 20ppm PFOS 100ppm PB 500ppm WY-14,643 50ppm 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 23 of 153 PFOS administration at 20ppm decreased the total liver content of DNA after 7 days on diet, whilst administration at 100ppm increased the total liver content of DNA after 1 and 7 days on diet, with a return to control levels after 28 days on diet for both dose levels. The administration of PB led to a sustained increase of approximately 20% at all time points examined, however administration of Wy14,643 led to marked increases of approximately 30% after 1 and 7 days on diet, and 60% after 28 days on diet (Table 12). Table 12. Total Liver DNA Content (mg DNA/whole liver) Days Liver DNA (mg/whole liver) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 21.42 2.06a 22.05 3.03 25.54 2.14*** 25.85 3.09** 27.78 2.39*** (100 9.6) (102.9 14.1) (119.2 10.0) (120.7 14.4) (129.7 11.2) 7 28.19 2.27 24.90 2.11** 30.81 2.51* 31.88 4.15* 36.60 3.28*** (100 8.1) (88.4 7.5) (109.3 8.9) (113.1 14.7) (129.9 11.6) 28 30.76 4.10 32.33 4.01 31.34 2.45 35.96 4.62* 47.89 9.40*** (100 13.3) (105.1 13.0) (101.9 8.0) (116.9 15.0) (155.7 30.6) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. 70 60 50 o o <5 30 O) E 20 Total liver DNA content (mg DNA/whole liver) Control PFOS 20ppm ________________________________________________ PFOS 100ppm PB 500ppm WY-14,643 50ppm 10 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 24 of 153 10.3.2 Peroxisome proliferation Administration of PFOS and Wy14,643 to rats resulted in marked increases in hepatic peroxisomal P-oxidation as determined by measurement of CN-insensitive palmitoyl CoA (PCO) oxidation. Both compounds increased PCO in a time dependent manner. Administration of PB, however, had no effect or a slight decrease in PCO (Table 13). Table 13. Hepatic Cyanide-insensitive Palmitoyl CoA Oxidation (PCO) Days on diet 0ppm PCO (nmol NAD+ reduced/minute/mg protein) PFOS PFOS PB 20ppm 100ppm 500ppm Wy14,643 50ppm 1 12.9 2.0a 14.0 1.5 15.1 1.7* 15.0 3.0 60.3 8.4*** (100 15.8) (108.0 11.6) (117.0 12.9) (116.3 23.1) (466.6 64.9) 7 15.9 2.9 15.4 2.2 20.9 2.7** 10.1 1.0*** 67.4 10.8*** (100 18.4) (96.6 14.0) (131.2 17.0) (63.3 6.1) (423.0 68.0) 28 14.7 2.2 20.2 3.2*** 55.3 6.2*** 12.8 2.2 115.4 11.9*** (100 15.2) (136.9 21.9) (376.0 41.9) (86.7 14.8) (783.9 80.7) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. PCoA Oxidation Control - PFOS 20ppm PFOS 100ppm PB 500ppm - W Y-14,643 50ppm 'o o Q. OE) E E <QZ o E 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 25 of 153 10.3.3 Total cytochrome P450 content of liver Administration of PFOS and PB to rats resulted in marked increases in hepatic total P450. Administration of PFOS increased total P450 in both a time, and dose dependent manner. Administration of Wy14,643, however, had no effect on hepatic total P450 content (Table 14). Table 14. Hepatic Total P450 Content Days Total P450 (nmol/mg protein) on 0ppm PFOS PFOS PB Wy-14,643 diet 20ppm 100ppm 500ppm 50ppm 1 0.71 0.15a 0.67 0.19 0.70 0.14 0.72 0.10 0.71 0.14 (100 20.9) (94.3 27.0) (98.2 19.1) (101.9 14.3) (100.1 19.2) 7 0.48 0.13 0.90 0.09*** 1.14 0.12*** 1.06 0.16*** 0.60 0.13 (100 26.7) (187.1 17.7) (238.4 24.3) (220.7 33.9) (124.0 26.0) 28 0.63 0.11 1.17 0.14*** 2.09 0.40*** 1.37 0.19*** 0.63 0.19 (100 17.7) (186.3 22.8) (333.4 63.8) (218.9 29.5) (100.6 29.4) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. 3.0 2.5 2.0 1.5 Q. 1.0 0.0 1 Day Microsomal Total P450 ----------------------------------------------------------------------------------------------------------------------------------- Control PFOS 20ppm PFOS 100ppm * * * PB 500ppm W Y -14,643 50ppm 7 Days Days on diet IE 28 Days CXR0481s Final Report Page 26 of 153 10.3.4 Laurie acid hydroxylation Lauric acid 12-hydroxylation (12-OH LAH) is used as a marker for cytochrome P450 4A activity. PFOS administration at 20ppm increased the 12-OH LAH by approximately 1.75fold after 7 days on diet, and 2-fold after 28 days on diet, whilst administration at 100ppm increased the 12-OH LAH by approximately 3-fold after 7 days on diet, and 4-fold after 28 days on diet. The administration of Wy14,643 led to very marked increases of approximately 6.5- fold, 20-fold and 25-fold after 1, 7 and 28 days on diet respectively. Administration of PB for up to 28 days, however, had essentially no effect on 12-OH LAH, with the exception of a 1.5- fold increase after 7 days on diet (Table 15). Table 15. Laurie Acid 12-Hydroxylation Days Lauric Acid 12 Hydroxylation (nmol/10minutes/mg protein) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 1.52 0.41a 1.59 0.53 1.45 0.39 1.19 0.42 9.96 4.49*** (100 26.8) (105.0 34.7) (95.3 25.4) (78.7 27.3) (656.0 295.8) 7 1.39 0.49 2.47 0.86** 4.21 1.27*** 2.15 0.50** 26.62 5.69*** (100 35.1) (178.1 62.1) (303.2 97.8) (155.1 36.3) (1918 409.8) 28 0.73 0.49 1.50 0.83* 3.05 1.56*** 0.61 0.42 18.71 8.54*** (100 67.7) (205.6 113.9) (418.7 214.6) (84.13 57.3) (2569 1173) a Values are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Microsomal Lauric Acid 12 Hydroxylation 35 Control PFOS 20ppm PFOS 100ppm 30 PB 500ppm W Y-14,643 50ppm 25 20 15 10 5 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 27 of 153 10.3.5 Pentoxyresorufin-O-depentylation Pentoxyresorufin-O-depentylation (PROD) is used as a marker for cytochrome P450 2B activity. PFOS administration at 20ppm initially decreased the PROD activity by approximately half after 1 and 7 days on diet, followed by a 2.5-fold increase after 28 days on diet. PFOS administration at 100ppm also initially decreased the PROD activity by approximately half after 1 day on diet, followed by a 5-fold, then 22-fold increase after 7 and 28 days on diet respectively. The administration of Wy14,643 showed no significant change in PROD activity after 1 day on diet, followed by a 2.5-fold, then 1.5-fold increase after 7 and 28 days on diet respectively. Administration of PB elicited very marked increases in PROD activity of approximately 11.5-fold, 162-fold and 118-fold after 1, 7 and 28 days on diet respectively (Table 16). Table 16. Pentoxyresorufin-O-depentylation Days on diet 0ppm PROD (pmols resorufin formed/minute/mg protein) PFOS PFOS PB 20ppm 100ppm 500ppm Wy14,643 50ppm 1 5.43 1.68a 2.97 0.93*** 2.89 0.66*** 62.2319.68*** 4.72 1.67 (100 31.0) (54.7 17.2) (53.2 12.1) (1147 363) (87.0 30.9) 7 3.39 1.23 1.86 0.64** 16.54 6.99*** 551 219*** 8.58 6.47* (100 36.4) (54.9 19.0) (488 206) (16272 6490) (253 191) 28 4.01 1.13 10.17 5.25** 88.96 27.55*** 473 131*** 6.64 3.56* (100 28.1) (253 131) (2218 687) (11784 3256) (165 88.8) a Values are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. CXR0481s Final Report Page 28 of 153 10.3.6 Testosterone hydroxylation Testosterone 6P-hydroxylation (Test-6P-OH) is used as a marker for cytochrome P450 3A activity. PFOS administration at 20ppm initially decreased the Test-6P-OH activity by approximately half after 1 day on diet, followed by a return to control levels after 7 days on diet, then a 1.5-fold increase after 28 days on diet. PFOS administration at 100ppm also initially decreased the Test-6P-OH activity by approximately half after 1 day on diet, followed by a 2-fold, then 2.5-fold increase after 7 and 28 days on diet respectively. Administration of PB led to a 3-fold then 4-fold increase in Test-6P-OH activity after 7 and 28 days on diet respectively. Administration of Wy14,643 for up to 28 days, however, had no effect on Test-6P-OH activity (Table 17). Table 17. Testosterone 6p hydroxylation Days Test-6P-OH (nmols 6P-OH formed/minute/mg protein on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 2.40 0.64a 1.54 0.46** 1.40 0.52*** 2.88 0.80 2.01 0.48 (100 25.6) (64.2 19.0) (58.2 21.5) (120.1 33.4) (83.6 19.8) 7 3.05 1.03 2.96 0.78 6.24 1.24*** 9.09 1.82*** 2.65 0.63 (100 33.9) (97.1 25.5) (204.6 40.6) (297.9 59.6) (86.8 20.6) 28 2.85 0.59 4.72 0.99*** 6.90 4.26** 10.99 2.26*** 3.24 0.64 (100 20.6) (165.8 34.9) (242.6 149.6) (386.3 79.4) (113.8 22.4) a Values are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. 14 10 s o Q. 05 o <> V> o E 0 Microsomal Testosterone 6beta Hydroxylation *** *** ** Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm -- it 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 29 of 153 10.4 Western Blotting The induction of several microsomal cytochromes P450 was demonstrated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting (Figure 1). SDS-PAGE and Western blotting was performed on hepatic microsomes obtained from 0ppm, PFOS 100ppm, PB 500ppm and Wy14,643 50ppm treated animals only. In each figure, the lane numbers refer to individual rat numbers as shown in Table 1 PFOS clearly induced CYP2B1/2, CYP2E1, CYP3A1 and CYP4A1. PB induced CYP2B1/2, CYP2E1 and CYP3A1, whilst Wy14,643 only induced CYP4A1. CXR0481s Final Report Page 30 of 153 Figure 1. SDS-PAGE and Western Blotting of Hepatic Microsomes Fig. 1A CYP2B1/2 CONTRO PFOS PR Wy-14 643 1 2 3 61 62 63 91 92 93 121 122 123 CONTROL PFOS PR Wy-14,643 4 5 6 64 65 66 94 95 96 124 125 126 CONTROI PFOS PR Wy-14 643 1 Day on diet CONTROL PFOS PR Wy-14,643 11 12 13 71 72 73 101 102 103 131 132 133 CONTROL PFOS PR Wy-14,643 14 15 16 74 75 76 104 105 106 134 135 136 CONTROL PFOS PR Wy-14,643 7 Days on diet 17 18 19 77 78 79 107 108 109 137 138 139 CONTROL PFOS PR Wy-14,643 21 22 23 81 82 83 111 112 113 141 142 143 CONTROL PFOS PR Wy-14,643 24 25 26 84 85 86 114 115 116 144 145 146 CONTROL PFOS PR Wy-14,643 28 Days on diet 27 28 29 87 88 89 117 118 119 147 148 149 0ppm, PFOS 100ppm, PB 500ppm and Wy14,643 50ppm treated animals only. CXR0481s Final Report Page 31 of 153 Fig. 1B CYP2E1 CONTROL PFOS PB Wy-14,643 " 1 2 3 61 62 63 91 92 93 121 122 123 CONTROL PFOS PB Wy-14.643 4 5 6 64 65 66 94 95 96 124 125 126 CONTROL PFOS PB Wy-14,643 1 Day on diet 7 8 9 67 68 69 97 98 99 127 128 129 CONTROI PFOS PB Wy-14.643 11 12 13 71 72 73 101 102 103 131 132 133 CONTROL PFOS PB Wy-14,643 14 15 16 74 75 76 104 105 106 134 135 136 CONTROL PFOS PB Wy-14,643 7 Days on diet 17 18 19 77 78 79 107 108 109 137 138 139 CONTROL PFOS PB Wy-14,643 21 22 23 81 82 83 111 112 113 141 142 143 CONTROL PFOS PB Wy-14,643 24 25 26 84 85 86 114 115 116 144 145 146 CONTROL PFOS PB Wy-14,643 28 Days on diet 27 28 29 87 88 89 117 118 119 147 148 149 0ppm, PFOS 100ppm, PB 500ppm and Wy14,643 50ppm treated animals only. CXR0481s Final Report Page 32 of 153 Fig. 1C CYP3A1 CONTROL PFOS PB Wv-14,643 1 2 3 61 62 63 91 92 93 121 122 123 CONTROL PFOS PB Wv-14,643 4 5 6 64 65 66 94 95 96 124 125 126 CONTROL PFOS PB Wy-14,643 1 Day on diet 7 8 9 67 68 69 97 98 99 127 128 129 CONTROL PFOS PB Wy-14,643 11 12 13 71 72 73 101 102 103 131 132 133 CONTROL PFOS PB Wy-14,643 14 15 16 74 75 76 104 105 106 134 135 136 CONTROL PFOS PB Wy-14,643 7 Days on diet 17 18 19 77 78 79 107 108 109 137 138 139 CONTROL PFOS PB Wy-14,643 21 22 23 81 82 83 111 112 113 141 142 143 CONTROL PFOS PB Wy-14,643 24 25 26 84 85 86 114 115 116 144 145 146 CONTROL PFOS PB Wy-14,643 28 Days on diet 27 28 29 87 88 89 117 118 119 147 148 149 0ppm, PFOS 100ppm, PB 500ppm and Wy14,643 50ppm treated animals only. CXR0481s Final Report Page 33 of 153 Fig. 1D CYP4A1 CONTROL PFOS PB Wv-14,643 1 2 3 61 62 63 91 92 93 121 122 123 CONTROL PFOS PB Wy-14,643 4 5 6 64 65 66 94 95 96 124 125 126 CONTROI PFOR PR W v -14.643 7 8 9 67 68 69 97 98 99 127 128 129 CONTROL PFOS PB Wv-14,643 11 12 13 71 72 73 101 102 103 131 132 133 CONTROL PFOS PB Wy-14,643 14 15 16 74 75 76 104 105 106 134 135 136 CONTROL PFOS PB Wv-14,643 1 Day on diet 7 Days on diet 17 18 19 77 78 79 107 108 109 137 138 139 CONTROL PFOS PB Wy-14,643 21 22 23 81 82 83 111 112 113 141 142 143 CONTROL PFOS PB Wy-14,643 24 25 26 84 85 86 114 115 116 144 145 146 CONTROL PFOS PB Wy-14,643 28 Days on diet 27 28 29 87 88 89 117 118 119 147 148 149 0ppm, PFOS 100ppm, PB 500ppm and Wy14,643 50ppm treated animals only. CXR0481s Final Report Page 34 of 153 10.5 Histopathology 10.5.1 Liver H&E Microscopically, treatment related findings were recorded in the liver. When compared with the control animals, a decrease in the usual glycogen induced hepatocellular vacuolation was noted in the animals of the 20ppm and 100ppm PFOS, 500ppm PB and 50ppm Wy14,643 groups, sacrificed after 28 days of treatment. In the 50ppm Wy14,643 group the decrease was already present after 1 and 7 days of treatment. Additionally, a minimal to slight, mainly centrilobular hepatocellular hypertrophy was noted, dose related in severity, in the animals of the 20ppm PFOS, 100ppm PFOS and 500ppm PB groups after 7 and 28 days of treatment. In the animals treated with 50ppm Wy14,643 a similar but mainly diffuse hepatocellular hypertrophy was recorded in all rats sacrificed after 1, 7 and 28 days of treatment. The severity of this finding was also increased with the duration of treatment. 10.5.2 Hepatic Apoptosis PFOS 100ppm, PB and Wy14,643 administration decreased the hepatic apoptotic index at all time points studied, reaching a minimum of 40%, 40% and 50% of control values respectively after 28 days of treatment (Table 18). Administration of PFOS 20ppm, however, resulted in a near 2-fold increase in apoptosis after 7 days of treatment, with a return to control values after 28 days of treatment. Table 18. Hepatic Apoptotic Indices Days Apoptotic Index (%) on 0ppm PFOS PFOS PB Wy14,643 diet 20ppm 100ppm 500ppm 50ppm 1 0.29 0.08a 0.28 0.16 0.23 0.08 0.14 0.04*** 0.10 0.05*** (100 28) (97 56) (78 29) (48 14) (33 16) 7 0.27 0.06 0.45 0.09*** 0.14 0.05*** 0.10 0.05*** 0.13 0.06*** (100 22) (164 32) (53 17) (35 18) (47 20) 28 0.25 0.06 0.24 0.07 0.10 0.03*** 0.10 0.05*** 0.13 0.05*** (100 23) (95 27) (40 12) (41 20) (51 22) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. CXR0481s Final Report Page 35 of 153 Hepatic Apoptotic Indices 0.6 Control PFOS 20ppm PFOS 100ppm 0.5 PB 500ppm W Y-14,643 50ppm 1 Day 7 Days Days on diet 28 Days 10.5.3 Cell proliferation in liver 20ppm PFOS, 100ppm PFOS, PB and Wy14,643 administration all increased the hepatocellular labelling index (S-phase) at all time points studied (Table 19). These increases were 2-, 3-, 6- and 10-fold, respectively, 1 day after commencement of treatment. After 7 days of treatment the levels of S-phase activity reached a maximum of 4-, 5-, 7- and 25-fold, respectively. By 28 days of treatment, the levels of S-phase activity reduced to 2-, 5-, 2- and 11-fold, respectively. Table 19. Hepatic S-phase Labelling Indices Days Labelling Index (%) on 0ppm diet PFOS 20ppm PFOS 100ppm PB 500ppm Wy14,643 50ppm 1 0.40 0.24a 0.82 0.32** 1.27 0.56*** 2.31 1.23*** 3.85 0.66*** (100 61) (204 81) (318 139) (577 306) (961 165) 7 0.64 0.20 2.62 1.70** 3.37 1.26*** 4.66 1.50*** 15.83 5.69*** (100 31) (411 265) (528 197) (730 236) (2479 891) 28 0.61 0.33 1.43 0.78** 3.20 1.25*** 1.48 0.55*** 6.49 2.83*** (100 55) (233 127) (523 204) (241 91) (1059 461) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group. A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. CXR0481s Final Report Page 36 of 153 Hepatic S-Phase 25 Control PFOS 20ppm PFOS 100ppm PB 500ppm W Y-14,643 50ppm 15 8 TCJ 05 o .TaO10 5 0 1 Day 7 Days Days on diet 28 Days 10.5.4 Thyroid H&E In the thyroid gland, no microscopic findings considered to be related to the treatment with the test items were recorded. 10.5.5 Thyroid in situ cell death The level of apoptosis found in all treatment groups, at all time points, was minimal. It was usual to find either no cells, or just one cell per thyroid, staining in the TUNEL assay as apoptotic, regardless of treatment. 10.5.6 Cell proliferation in thyroid Wy14,643 administration increased the thyroid follicular cell labelling index (S-phase) after 1 and 7 days of treatment, whilst PB administration increased the level of S-phase activity only after 7 days of treatment (Table 20). After 28 days of treatment the levels of S-phase activity returned to control values in both Wy14,643 and PB treated animals. PFOS 20ppm and PFOS 100ppm treatment had no effect on S-phase at any time point studied. CXR0481s Final Report Page 37 of 153 Table 20. Thyroid S-phase Labelling Indices Days Labelling Index (%) on 0ppm PFOS1 PFOS1 P B Wy14,6431 diet 20ppm 100ppm 500ppm 50ppm 1 0.92 0.27a 0.85 0.58 0.78 0.60 0.99 0.37 1.42 0.44* (100 29) (92 64) (84 65) (108 40) (154 48) 7 2.32 1.09 2.87 1.15 2.89 0.59 7.24 3.43** 5.61 2.06** (100 47) (124 49) (124 25) (311 148) (241 89) 28 1.81 0.74 1.92 0.28 1.35 0.33 2.03 0.84 1.55 0.47 (100 41) (106 15) (74 18) (112 46) (86 26) aValues are Mean SD. Values in parenthesis are mean % control SD. n = 10 per group; 1n = 5 per group A Student's t-test (2-sided) was performed on the results; *statistically different from control p<0.05; ** p<0.01; *** p<0.001. Thyroid S-Phase 12 Control PFOS 20ppm PFOS 100ppm PB 500ppm 10 W Y-14,643 50ppm 8 6 4 2 0 1 Day 7 Days Days on diet 28 Days CXR0481s Final Report Page 38 of 153 11. DISCUSSION In this study both Wy14,643 and phenobarbital exhibited their characteristic hepatic effects. These agents also had minimal effects in the thyroid follicular cells increasing S-phase by 2 3-fold after 7days administration. Wy14,643 induced hepatomegaly that was characterised by peroxisome proliferation, selective induction of CYP4A1, inhibition of apoptosis and stimulation of cell proliferation as determined by increased DNA synthesis. Marked decreases in plasma cholesterol and triglycerides were also seen; again characteristic of a peroxisome proliferator. Phenobarbital also induced hepatomegaly but was not a peroxisome proliferator, however it did exhibit the prototypical effects of a (not surprisingly) "phenobarbital-like" enzyme inducer. That is, the liver enlargement was characterised by induction of CYP2B and CYP3A, inhibition of apoptosis and stimulation of cell proliferation demonstrated by an increased labelling index. The peroxisome proliferation, the enzyme induction and the effects of these compounds on apoptosis and S-phase in the liver are transcriptionally regulated via members of the nuclear hormone receptor super family (e.g. PPARa, CAR and PXR). These receptors play vital roles in the regulation of both intermediary metabolism, xenobiotic metabolism and in the regulation of liver growth. Evidence exists to suggest that the long-term activation of these receptors by non-genotoxic carcinogens is the initial step in such hepatocarcinogenesis. In this study, PFOS appeared to exhibit the combined effects of PB and Wy14,643, thus behaving as a combined peroxisome proliferator and "phenobarbital-like" enzyme inducer. Thus, these data in totality suggest that PFOS may activate at least three nuclear receptors: PPARa, CAR and PXR. 12. DATA STORAGE AND ARCHIVING The data has been held in a Study File according to CXR Biosciences SOPs 0003 and 0005. On completion of the study, the following items will be retained in the CXR Archive: Study protocol and possible amendments Study Report and possible amendments Study File (including Study Diary) Relevant correspondence All items will be kept for two years from the date of signing the Study Report unless, in the opinion of CXR Biosciences, the quality of the specimens no longer allows evaluation. At the end of the two-year period, the Study Sponsor will be contacted and the items either sent to the Study Sponsor or retained by CXR Biosciences under a separate agreement. CXR0481s Final Report Page 39 of 153 13. REPORT The Study Report has been prepared as described in CXR Biosciences SOP 0004. 14. QUALITY ASSURANCE Study activities at the Test Facility (CXR Biosciences Ltd and BSRU) have been conducted according to relevant SOPs. There were no deviations from this protocol. No claim of GLP compliance has been made for this study. 15. STUDY DIRECTORS STATEMENT Final Report signature by the Study Director indicates acceptance of responsibility for the validity and integrity of the Study data. CXR0481s Final Report Page 40 of 153 APPENDIX 1. PATHOLOGY REPORT CXR0481s Final Report Page 41 of 153 PATHOLOGY REPORT TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. CXR : 0481/s PATHOL. NO. 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 PREPARED BY: Dr. med. vet. Ortwin Vogel Veterinary Pathologist CXR0481s Final Report Page 42 of 153 PATHOLOGY REPORT TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TABLE OF CONTENTS AUTHENTICATION PAGE CXR I 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 PAGE : 1 PRINCIPAL SECTION SUMMARY METHODS RESULTS CONCLUSIONS REFERENCES 2- 3 4- 5 6- 7 8- 9 10 EXPLANATION OF CODES AND SYMBOLS SUMMARY TABLES SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX NECROPSY STATUS: INTERIM SACRIFICE GROUP (K1) SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX NECROPSY STATUS: INTERIM SACRIFICE GROUP (K2) SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX NECROPSY STATUS: INTERIM SACRIFICE GROUP (K3) INDIVIDUAL ANIMAL DATA TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) ANIMAL HEADING DATA DOSE GROUP 01 TEXT OF MICROSCOPIC FINDINGS DOSEGROUP 01 ANIMAL HEADING DATA DOSE GROUP 02 TEXT OF MICROSCOPIC FINDINGS DOSEGROUP 02 11 12 13 14 15 - 29 30 31 - 45 46 47 - 61 CXR0481s Final Report Page 43 of 153 PATHOLOGY REPORT TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TABLE OF CONTENTS ANIMAL HEADING DATA DOSE GROUP 03 TEXT OF MICROSCOPIC FINDINGS DOSE GROUP 03 ANIMAL HEADING DATA DOSE GROUP 04 TEXT OF MICROSCOPIC FINDINGS DOSE GROUP 04 ANIMAL HEADING DATA DOSE GROUP 05 TEXT OF MICROSCOPIC FINDINGS DOSE GROUP 05 PAGE CXR II 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 PAGE : 62 63 - 77 78 79 - 93 94 95 - 109 CXR0481s Final Report Page 44 of 153 PATHOLOGY REPORT PAGE CXR 1/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 AUTHENTICATION The undersigned hereby declares that the histopathology data in this re port were compiled by him, and that they reflect accurately the raw data records. Dr. med. vet. Ortwin Vogel Veterinary Pathologist TPC Toxicologic Pathology Consultancy Goethestr. 26 D-24116 Kiel CXR0481s Final Report Page 45 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. SUMMARY PAGE CXR 2/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 To evaluate possible toxic effects of the test item "PFOS", pathomorphological examinations were performed on 150 male Sprague Dawley rats. The rats were allocated to 5 dose groups each consisting of 30 males and received the test items via the feed at dose levels of 20 ppm "PFOS", 100 ppm "PFOS", 500 ppm "PB" or 50 ppm "Wyeth 14,643" (groups 02, 03, 04 and 05, respectively) for a treatment period of 1 (sacrifice K1), 7 (sacri fice K2) or 28 days (sacrifice K3). The rats of group 01 received the same diet without any test item and served as control animals. At the end of the treatment periods, the rats were sacrificed and detailed necropsies performed. Histopathological examination was performed on the liver and thyroid from all animals of all groups. All animals survived the duration of the study. Microscopically, treatment related findings were recorded in the liver. When compared with the control animals, a decrease in the usual glycogen induced hepatocellular vacuolation was noted in the animals of groups 02 (20 ppm PFOS), 03 (100 ppm PFOS), 04 (500 ppm PB) and 05 (50 ppm Wyeth 14.643), sacrificed after 28 days of treatment. In group 05 (50 ppm Wyeth 14.643) the decrease was already present after 1 and 7 days of treatment. In addition, a minimal to slight, mainly centrilobular hepatocellular hypertrophy was noted, dose related in severity, in the animals of groups 02 - 04 after 7 and 28 days of treatment. In the animals treated with 50 ppm Wyeth 14,643 (group 05), a similar but mainly diffuse hepatocellular hypertrophy was recorded in all rats sacrificed after 1, 7 and 28 days of treatment. The severity of this finding was also increased with the dura tion of treatment. In the thyroid gland, no microscopic finding was recorded, that is con sidered to be related to the treatment with the test items. All other microscopic findings recorded did not distinguish significantly treated rats from vehicle control rats or the differences noted were regarded as random effects. All these findings are considered to be spon taneous in nature and within the normal background pathology commonly seen in rats of these strains and age. CXR0481s Final Report Page 46 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. SUMMARY PAGE CXR 3/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 Under the conditions of this study, the repeated administration of 20 or 100 ppm "PFOS", 500 ppm "PB" or 50 ppm "Wyeth 14,643" via the feed to rats of the Sprague Dawley strain for a treatment period of up to 28 days produced morphological evidence of a liver toxicity. CXR0481s Final Report Page 47 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding CXR Biosciences Ltd. METHODS PAGE CXR 4/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 Group Design for Pathological Evaluation Dose Group 01 02 03 04 05 Pathdata Test item* Group Number of Rats * Males 01 0 (Control) 10** 10*** 10**** 02 20 ppm PFOS 10** 10*** 10**** 03 100 ppm PFOS 10** 10*** 10**** 04 500 ppm PB 10** 10*** 10**** 05 50 ppm Wyeth 14,643 10** 10*** 10**** Animal numbers Males 1 - 10 11 - 20 21 - 30 31 - 40 42 - 50 51 - 60 61 - 70 71 - 80 81 - 90 91 - 100 101 - 110 111 - 120 121 - 130 131 - 140 141 - 150 * strain: Sprague Dawley (CD),(Breeder: Charles River, UK); ** These animals are listed in the tables under necropsy status K1. *** These animals are listed in the tables under necropsy status K2. **** These animals are listed in the tables under necropsy status K3. Administration of the Test Item The in-life part of the study was performed at CXR Biosciences Ltd., James Lindsay Place, Dundee DD1 5JJ, UK. The animals were administered either 20 ppm PFOS (group 02), 100 ppm PFOS (group 03), 500 ppm PB (group 04 or 50 ppm Wyeth 14,643 (group 05) in the diet ad libitum for either 1 (sacrifice K1), 7 (sacrifice K2) or 28 days (sacrifice K3). The animals of group 01 received the same diet without the test items and served as control animals. CXR0481s Final Report Page 48 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. METHODS PAGE CXR 5/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 Necropsy and Histopathology At the end of the assigned study periods (1 day, (K1), 7 days (K2) or 28 days (K3) after administration), the rats were killed by exposure to a rising concentration of CO2. Two samples of the liver (one from the left lobe, one from the median lobe) and the thyroid including parathyroid gland together with trachea and oesophagus were taken and preserved in 10% neutral buffered formaldehyde (NBF). Histotechnique was performed at CXR Biosciences Ltd., UK. The preserved liver and thyroid samples of all rats of all groups were trimmed, processed, embedded in paraffin wax and sectioned at a nominal thickness of about 5 pm. All sections were stained with haematoxylin and eosin. One section of each organ sample was examined by the undersigned pathologist by light microscope. Data compilation The animal heading data and necropsy findings recorded by CXR at post mortem examination were taken from the post-mortem records and entered manually into the PathData Computer system, current version, under Patho logy No. 07041 (PathData is a registered trademark of Pathology Data Sys tems, Inc.). Microscopic findings are recorded by the undersigned patho logist during histopathological examination using on-line data input into the pathology computer system. All microscopic observations are presented for each rat in the "Table of Individual Microscopic Findings" as well as in full descriptive terms in the "Individual Animal Data - Text of Gross and Microscopic Findings". The incidence of microscopic findings is also presented in tabular form. Incidence tables were created by the Pathdata software. Histologic alter ations were described, wherever possible, according to their distribu tion, severity and morphologic character. Severity scores were assigned as given under "Explanation of Codes and Symbols". The slides were evaluated during the period July 16 - 28, 2007. Archive For archiving purposes, the raw data including a CD-ROM containing the electronic study data and the microscopic slides will be returned to CXR Biosciences Ltd., James Lindsay Place, Dundee DD1 5JJ, UK. CXR0481s Final Report Page 49 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS) RAT, 28 days, feeding CXR Biosciences Ltd. RESULTS PAGE CXR 6/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 Mortality All animals survived the duration of the study. Microscopic findings At histopathologic examination, few microscopic findings were recorded. These findings are described in detail in the "Individual Animal Data Text of Gross and Microscopic Findings". Liver: Sacrifice after 1 day of treatment: A slight to moderate, periportal hepatocellular vacuolation that is con sidered to be due to an intracytoplasmic glycogen accumulation was promi nent in the control animals of group 01. Hepatocellular vacuolation was also present, similar in incidence and severity, in the animals of groups 02 (20 ppm PFOS), 03 (100 ppm PFOS) and 04 (500 ppm PB). In group 05 (50 ppm Wyeth 14,643), this kind of vacuolation occurred, minimal in degree, in 1 rat only. In contrast, a slight, mainly diffuse hepatocellular hypertrophy, that was not observed in the animals of groups 01 - 04, was recorded in all animals of this group instead. Sacrifice after 7 days of treatment: A similar hepatocellular vacuolation as recorded in the liver of the ani mals sacrificed after 1 day of treatment was also noted, minimal to mark ed in degree, in the control and treated animals of groups 01 - 04. A minimal hepatacellular vacuolation occurred also in 3 animals of group 05 (50 ppm Wyeth 14,643). In addition, hepatocellular hypertrophy that did not occur in control animals was noted in all treated groups. Dose dependent in severity, this mainly centrilobular finding was observed in 7 animals of group 02 (20 ppm PFOS, mean severity 1.0) and all animals of groups 03 (100 ppm PFOS, mean severity 1.4) and 04 (500 ppm PB, mean severity 2.0). In group 05 (50 ppm Wyeth 14,643) a mainly diffuse hepato cellular hypertrophy was found in all animals with a mean severity of 2.7). Sacrifice after 28 days of treatment: Hepatocellular vacuolation similar to the finding noted in the animals after 1 and 7 days of treatment was recorded also in all control animals. A decrease in incidence and severity of this finding was noted in groups 02 - 05. In contrast, hepatocellular hypertrophy was noted the animals of these groups. This alteration occurred, minimal to moderate in degree, in CXR0481s Final Report Page 50 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. RESULTS PAGE CXR 7/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 all animals of group 02 (20 ppm PFOS, mean severity 2.1), 03 (100 ppm, PFOS, mean severity 2.0) and 04 (500 ppm PB, mean severity 2.7). In group 05 (50 ppm Wyeth 14,643) a mainly diffuse hepatocellular hypertrophy was found in all animals with a mean severity of 2.9. Thyroid: There were no microscopic findings recorded that are considered to be related to the treatment with any test items in either sacrifice group. The type, incidence and severity of all other microscopic alterations noted did not indicate a relationship to the treatment with the test item or the differences noted were regarded as random effects. These findings were regarded to be spontaneous in nature and within the normal back ground pathology commonly seen in rats of these strains and age. CXR0481s Final Report Page 51 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS) RAT, 28 days, feeding CXR Biosciences Ltd. CONCLUSIONS PAGE CXR 8/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 In this 28-day toxicity study of the test item "PFOS" administered with the feed to Sprague Dawley rats at dose levels of 0 ppm (control), 20 ppm PFOS, 100 ppm PFOS, 500 ppm Pb or 50 ppm Wyeth 14,643, no premature death occurred. A similar incidence and severity of hepatocellular vacuolation, occurred in rats of groups 01 - 04 sacrificed after 1 or 7 days of treatment. In the animals of group 05 (50 ppm Wyeth 14,643), the degree of vacuolation was markedly decreased. After 28 days of treatment, a similar vacuolation (mean severity 2.6) was also recorded in the control animals. In the treated animals of groups 02 - 05, a decrease in the degree of this vacu olation was recorded (mean severity 1.8, 1.0, 1.4, 1.0). This kind of vacuolation is considered to be due to the intracytoplasmic presence of glycogen and is quite commonly observed in well-fed, untreated rodents. The difference between control and treated animals noted in this study after 1 day (group 05), 7 (group 05) and 28 days of treatment (groups 02 - 05) is considered to be related to the treatment with the test items. A slight, mainly centrilobular hepatocellular hypertrophy was noted, dose related in severity, in the animals of groups 02 - 04 after 7 days of treatment and was increased in severity after 28 days of treatment. In the animals treated with 50 ppm Wy-14,643, a similar but mainly diffuse hepatocellular hypertrophy was recorded in all rats sacrificed after 1, 7 and 28 days of treatment. The severity of this finding was also increased with the duration of treatment. In the absence of any hepatocellular hypertrophy in the control animals, this alteration is considered to be related to the treatment with the various test items. A large number of medicinal agents with different chemical structures and therapeutic activities produce liver enlargement when given in high doses to species used in toxicity studies (Greaves, 1990). This enlargement that may be characterized morphologically by hepatocyte hypertrophy, hepatocyte hyperplasia or both may be accompanied by an increase in activity of the hepatic microsomal drug metabolizing enzymes in the absence of any morphogical evidence of hepatocellular damage. The changes are typically reversible on cessation of treatment. There were no microscopic findings recorded in the thyroid gland that are considered to be related to the treatment with any test items in either sacrifice group. CXR0481s Final Report Page 52 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. CONCLUSIONS PAGE CXR 9/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 All other microscopic findings recorded did not distinguish significantly treated rats from vehicle control rats or the differences noted were regarded as random effects. All these findings are considered to be spon taneous in nature and within the normal background pathology commonly seen in rats of these strains and age. Under the conditions of this study, the repeated administration of 20 or 100 ppm "PFOS", 500 ppm "PB" or 50 ppm "Wyeth 14,643 via the feed to rats of the Sprague Dawley strain for a treatment period of up to 28 days pro duced morphological evidence of a liver toxicity. CXR0481s Final Report Page 53 of 153 PATHOLOGY REPORT PRINCIPAL SECTION TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. REFERENCES PAGE CXR 10/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 Greaves P. (1990): Histopathology of Preclinical Toxicity Studies: Interpretation and Relevance in Drug Safety Evaluation. Digestive System I, 278-392. Elsevier, Amsterdam New York Oxford CXR0481s Final Report Page 54 of 153 PATHOLOGY REPORT TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. EXPLANATION OF CODES AND SYMBOLS PAGE CXR 11/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 CODES AND SYMBOLS USED AT ANIMAL LEVEL: M = Male animal K1...K9 = Interim sacrifice groups 1...9 CODES AND SYMBOLS USED AT ORGAN LEVEL: ' = Histologic examination not required + = Organ examined, findings present - = Organ examined, no pathologic findings noted (AOFT only) ( = Only one of paired organs examined/present CODES AND SYMBOLS USED AT FINDING LEVEL: GRADE 1 GRADE 2 GRADE 3 GRADE 4 P ( Minimal / very few / very small Slight / few / small Moderate / moderate number / moderate size Marked / many / large Finding present, severity not scored Finding unilateral in paired organs CXR0481s Final Report Page 55 of 153 PATHOLOGY REPORT SUMMARY TABLES TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS) RAT, 28 days, feeding CXR Biosciences Ltd. PAGE CXR 12/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX Necropsy Status: INTERIM SACRIFICE GROUP (K1) Sex Males Dose Group No. Animals per Dose Group 01 02 03 04 10 10 10 10 LIVER No Examined 10 10 10 10 - mononuclear cell GRADE 1 8 4 7 8 infiltrate(s) TOTAL AFFECTED 8 4 7 8 MEAN SEVERITY 1.0 1.0 1.0 1.0 - (mixed) inflammatory GRADE 1 8 7 5 4 cell infiltration TOTAL AFFECTED 8 7 5 4 MEAN SEVERITY 1.0 1.0 1.0 1.0 - vacuolation GRADE 1 _ _ _ _ GRADE 2 6 3 5 4 GRADE 3 4 7 5 5 TOTAL AFFECTED 10 10 10 9 MEAN SEVERITY 2.4 2.7 2.5 2.6 - hypertrophy GRADE 2 _ _ _ _ TOTAL AFFECTED _ _ _ _ MEAN SEVERITY _ _ _ _ THYROID GLAND No. Examined 10 10 10 10 - branchiogenic cyst GRADE 1 _ _ _ 1 TOTAL AFFECTED _ _ _ 1 MEAN SEVERITY _ _ _ 1.0 - cyst GRADE 1 _ _ _ 1 TOTAL AFFECTED _ _ _ 1 MEAN SEVERITY _ _ _ 1.0 - mononuclear cell GRADE 1 _ 1 _ _ infiltrate(s) TOTAL AFFECTED _ 1 _ _ MEAN SEVERITY _ 1.0 _ _ 05 10 10 7 7 1.0 8 8 1.0 1 _ _ 1 1.0 10 10 2.0 10 _ _ _ _ _ _ 1 1 1.0 CXR0481s Final Report Page 56 of 153 PATHOLOGY REPORT SUMMARY TABLES TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS) RAT, 28 days, feeding CXR Biosciences Ltd. PAGE CXR 13/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX Necropsy Status: INTERIM SACRIFICE GROUP (K2) Sex Males Dose Group No. Animals per Dose Group 01 02 03 04 10 10 10 10 LIVER No Examined 10 10 10 10 - mononuclear cell GRADE 1 3 4 2 1 infiltrate(s) TOTAL AFFECTED 3 4 2 1 MEAN SEVERITY 1.0 1.0 1.0 1.0 - (mixed) inflammatory GRADE 1 7 3 8 9 cell infiltration TOTAL AFFECTED 7 3 8 9 MEAN SEVERITY 1.0 1.0 1.0 1.0 - vacuolation GRADE 1 _ _ _ 1 GRADE 2 4 2 5 7 GRADE 3 5 8 5 2 GRADE 4 1 _ _ _ TOTAL AFFECTED 10 10 10 10 MEAN SEVERITY 2.7 2.8 2.5 2.1 - hypertrophy GRADE 1 GRADE 2 GRADE 3 TOTAL AFFECTED MEAN SEVERITY _76_ _ _ 4 10 ____ _ 7 10 10 _ 1.0 1.4 2.0 THYROID GLAND No. Examined 10 10 10 10 - branchiogenic cyst GRADE 1 3 1 1 2 TOTAL AFFECTED 3 1 1 2 MEAN SEVERITY 1.0 1.0 1.0 1.0 - mononuclear cell GRADE 1 _ 2 _ _ infiltrate(s) TOTAL AFFECTED _ 2 _ _ MEAN SEVERITY _ 1.0 _ _ 05 10 10 1 1 1.0 8 8 1.0 3 _ _ _ 3 1.0 _ 3 7 10 2.7 10 1 1 1.0 _ _ _ CXR0481s Final Report Page 57 of 153 PATHOLOGY REPORT SUMMARY TABLES TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS) RAT, 28 days, feeding CXR Biosciences Ltd. PAGE CXR 14/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 SUMMARY INCIDENCE OF GRADINGS BY ORGAN/GROUP/SEX Necropsy Status: INTERIM SACRIFICE GROUP (K3) Sex Males Dose Group No. Animals per Dose Group 01 02 03 04 10 10 10 10 LIVER No Examined 10 10 10 - mononuclear cell GRADE 1 3 2 5 infiltrate(s) TOTAL AFFECTED 3 2 5 MEAN SEVERITY 1.0 1.0 1.0 10 - - - (mixed) inflammatory GRADE 1 10 6 4 9 cell infiltration TOTAL AFFECTED 10 6 4 9 MEAN SEVERITY 1.0 1.0 1.0 1.0 - fatty change (vacuolation) - vacuolation GRADE 2 TOTAL AFFECTED MEAN SEVERITY GRADE 1 GRADE 2 GRADE 3 - _ _ 4 6 _3 _3 _ 2.0 33 41_ - - 5 4 - TOTAL AFFECTED MEAN SEVERITY 10 2.6 83 9 1.0 1.4 CO <--1 - hypertrophy GRADE 1 GRADE 2 GRADE 3 TOTAL AFFECTED MEAN SEVERITY _1-- 7 10 3 _2-7 _ 10 10 10 _ 2.1 2.0 2.7 THYROID GLAND No. Examined 10 10 10 10 - branchiogenic cyst GRADE 1 2 1 1 1 TOTAL AFFECTED 2 1 1 1 MEAN SEVERITY 1.0 1.0 1.0 1.0 05 10 10 1 1 1.0 5 5 1.0 - - 8 - 8 1.0 1 9 10 2.9 10 1 1 1.0 CXR0481s Final Report Page 58 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 15/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days , feeding : CXR Biosciences Ltd. PATHOL NO.: 07041 VOO DATE 12-OCT -07 PathDataSystem V6. 2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 01, Control ANIMAL NUMBER : 1 2 3 4 5 6 7 8 9 10 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . ++++++++++ 1. 1. . 1 . 1. . 1. 1. 1. 1. . 1. 1. . 1. 1. 1. 1. 1. 1. 3. 3. 2. 2 . 3. 2. 2. 3. 2. 2. THYROID GLAND - ectopic thymus --+ . . P. - (- - - - + . . . . . ( P. . CXR0481s Final Report Page 59 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 16/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT -07 PathDataSystem V6. 2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 01, Control ANIMAL NUMBER : 11 12 13 14 15 16 17 18 19 20 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . ++ . 1. 1. 1. 4. 3. ++++ . . . 1. 1. 1 . . 1. 3. 3 . 3. 2. ++++ . . . 1. . . 1. 1. 3. 2. 2. 2. THYROID GLAND - ectopic thymus - branchiogenic cyst (+ - - - - + . . . . . ( P. ( 1. . . . . ( 1. -+-- .... . ( 1. .. CXR0481s Final Report Page 60 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 17/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS RAT, 28 days, feeding CXR Biosciences Ltd. PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP 01, Control ANIMAL NUMBER 21 22 23 24 25 26 27 28 29 30 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. ++++ . . . 1. 1. 1. 1. 1. 3. 3. 2. 3. ++ . 1. 1. 1. 3. 2. ++ . 1. 1. 1. 3. 2. ++ .. 1. 1. 2. 3. THYROID GLAND - ectopic thymus - branchiogenic cyst ---++--+-- . . . ( P. . . . ( P. .. . . . 1. ( 1. ..... CXR0481s Final Report Page 61 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 18/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days , feeding : CXR Biosciences Ltd. PATHOL NO.: 07041 VOO DATE 12-OCT -07 PathDataSystem V6. 2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 02, PFOS 20 ppm ANIMAL NUMBER : 31 32 33 34 35 36 37 38 39 40 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . ++++++++++ 1. . . 1 . . . . 1. . 1. . 1. 1. 1 . 1. 1. . . 1. 1. 3. 3. 3. 3 . 2. 2. 3. 3. 2. 3. THYROID GLAND - mononuclear infilt . --+------- . . ( 1. ....... CXR0481s Final Report Page 62 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 19/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS RAT, 28 days, feeding CXR Biosciences Ltd. PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP 02, PFOS 20 ppm ANIMAL NUMBER 41 42 43 44 45 46 47 48 49 50 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. - hypertrophy, hepat. ++++++++++ 1. 1. 1. 1. 1. 1. 1. 3. 3. 3. 3. 3. 2. 3. 3. 2. 3. 1. 1. 1. 1. 1. 1. 1. THYROID GLAND - branchiogenic cyst - mononuclear infilt. - + - - - - + + (- ( 1. ( 1. ( 1. CXR0481s Final Report Page 63 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 20/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR Perfluorooctane Sulfonate (PFOS RAT, 28 days, feeding CXR Biosciences Ltd. PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP 02, PFOS 20 ppm ANIMAL NUMBER 51 52 53 54 55 56 57 58 59 60 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. - hypertrophy, hepat. ++++++++++ 1. 1. 1. 1. 1. 1. 1. 1. 2. 1. 1. 2. 2. 1. 2. 3. 2. 2. 2. 3. 2. 2. 2. 2. 3. 1. THYROID GLAND +----+---- - ectopic thymus ( P. - branchiogenic cyst ( 1. CXR0481s Final Report Page 64 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 21/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days , feeding : CXR Biosciences Ltd. PATHOL NO.: 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 03, PFOS 100 ppm ANIMAL NUMBER : 61 62 63 64 65 66 67 68 69 70 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . ++++++++++ 1. 1. 1. 1 . 1. 1. 1. ... . 1. 1. 1 . . . . 1. 1. . 3. 2. 2. 3 . 2. 3. 3. 3. 2. 2. THYROID GLAND ---------- CXR0481s Final Report Page 65 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 22/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT -07 PathDataSystem V6. 2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 03, PFOS 100 ppm ANIMAL NUMBER : 71 72 73 74 75 76 77 78 79 80 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . - hypertrophy, hepat . ++++++++++ 1. . . 1. . . . . . . . 1. 1. . 1. 1. 1. 1. 1. 1. 3. 3. 3. 3. 2. 3. 2. 2. 2. 2. 1. 1. 1. 1. 1. 1. 2. 2. 2. 2. THYROID GLAND - branchiogenic cyst +--------- ( 1. ......... CXR0481s Final Report Page 66 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR : 23/ 109 : 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 03, PFOS 100 ppm ANIMAL NUMBER : 81 82 83 84 85 86 87 88 89 90 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 LIVER - mononuclear infilt. - inflammatory cell - fatty change - vacuolation, hepat. - hypertrophy, hepat. ++++++++++ 1. 1. 1. 1. 1. 1. 1. 1. 1. 2. 2. 2. 1. 1. 1. 2. 2. 2. 2. 2. 2. 2. 2. 2. 2. THYROID GLAND - branchiogenic cyst ---------+ ( 1. CXR0481s Final Report Page 67 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 24/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 04, PB 500 ppm ANIMAL NUMBER : 91 92 93 94 95 96 97 98 99 100 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. +++++++++- 1. 1. 1. 1. 1. 1. 1. 1. . 1. 1. 1. 1. . 3. 3. 3. 3. 3. 2. 2. 2. 2. . THYROID GLAND + - - (- - - - + - - - ectopic thymus ( P. - branchiogenic cyst ( 1. - cyst ( 1. CXR0481s Final Report Page 68 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 25/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL NO.: 07041 VOO DATE 12-OCT-07 PathDataSystem V6 .2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 04, PB 500 ppm ANIMAL NUMBER : 101 102 103 104 105 106 107 108 109 110 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 LIVER - mononuclear infilt . - inflammatory cell - vacuolation, hepat . - hypertrophy, hepat . ++++++++++ . 1. ........ 1. . 1. 1 . 1. 1. 1. 1. 1. 1. 2. 3. 1. 2 . 2. 2. 2. 3. 2. 2. 2. 2. 2. 2 . 2. 2. 2. 2. 2. 2. THYROID GLAND - branchiogenic cyst - - - (- - - - + (+ - . . . . . . . ( 1. ( 1. . CXR0481s Final Report Page 69 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 26/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6 .2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 04, PB 500 ppm ANIMAL NUMBER : 111 112 113 114 115 116 117 118 119 120 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 LIVER - inflammatory cell - vacuolation, hepat . - hypertrophy, hepat . ++++++++++ 1. 1. 1. 1 . . 1. 1. 1. 1. 1. 2. 1. 1. . 2. 1. 2. 1. 2. 1. 3. 2. 3. 3 . 2. 3. 2. 3. 3. 3. THYROID GLAND - branchiogenic cyst ---------+ . . . . . . . . . ( 1. CXR0481s Final Report Page 70 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 27/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 05, Wy-14,634 50 ppm ANIMAL NUMBER : 121 122 123 124 125 126 127 128 129 130 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 MK1 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. - hypertrophy, hepat. ++++++++++ 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 1. 2. 2. 2. 2. 2. 2. 2. 2. 2. 2. THYROID GLAND - ectopic thymus - mononuclear infilt. ------+-+( P. ( 1. CXR0481s Final Report Page 71 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 28/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 05, Wy-14,634 50 ppm ANIMAL NUMBER : 131 132 133 134 135 136 137 138 139 140 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 MK2 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. - hypertrophy, hepat. THYROID GLAND - branchiogenic cyst ++++++++++ . . . . 1. ..... 1. 1. . 1. 1. 1. 1. . 1. 1. 1. 1. 1. . . . . . . . 2. 2. 2. 3. 3. 3. 3. 3. 3. 3. ------+--......................... ( 1. CXR0481s Final Report Page 72 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA PAGE CXR 29/ 109 0481/s TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) DOSE GROUP : 05, Wy-14,634 50 ppm ANIMAL NUMBER : 141 142 143 144 145 146 147 148 149 150 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 MK3 LIVER - mononuclear infilt. - inflammatory cell - vacuolation, hepat. - hypertrophy, hepat. THYROID GLAND - branchiogenic cyst ++++++++++ . . . . . 1. .... . 1. 1. . . . 1. 1. . 1. 1. 1. 1. 1. 1. 1. . . 1. 1. 3. 3. 3. 3. 3. 3. 3. 3. 3. 2. --+------. . ( 1. CXR0481s Final Report Page 73 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. ANIMAL HEADING DATA DOSE GROUP : 01, Control PAGE CXR 30/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 ANIMAL SEX NUMBER M/F 1M 2M 3M 4M 5M 6M 7M 8M 9M 10 M 11 M 12 M 13 M 14 M 15 M 16 M 17 M 18 M 19 M 20 M 21 M 22 M 23 M 24 M 25 M 26 M 27 M 28 M 29 M 30 M DEFINED AND FINAL STATE OF NECROPSY K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 TEST DAYS FIRST AND LAST DAY UNDER TEST DATE OF NECROPSY 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 CXR0481s Final Report Page 74 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 31/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 1 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 2 CXR0481s Final Report Page 75 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 32/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 3 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, bilateral * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 4 CXR0481s Final Report Page 76 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 33/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 5 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 6 CXR0481s Final Report Page 77 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 34/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 7 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 8 CXR0481s Final Report Page 78 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 35/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 9 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 10 CXR0481s Final Report Page 79 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 36/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 11 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 4 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present -branchiogenic cyst, unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 12 CXR0481s Final Report Page 80 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 37/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 13 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 14 CXR0481s Final Report Page 81 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 38/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 15 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral -branchiogenic cyst, unilateral, grade 1 16 CXR0481s Final Report Page 82 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 39/ 109 0481/s PATHOL. NO. 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 17 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 **ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 18 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 19 CXR0481s Final Report Page 83 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 40/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 20 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 21 CXR0481s Final Report Page 84 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 41/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 22 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 23 CXR0481s Final Report Page 85 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 42/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 24 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral -branchiogenic cyst, bilateral, grade 1 * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 25 CXR0481s Final Report Page 86 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 43/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 26 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 27 CXR0481s Final Report Page 87 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 44/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 28 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 29 CXR0481s Final Report Page 88 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 01, Control PAGE CXR 45/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 30 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 89 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. ANIMAL HEADING DATA DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 46/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 ANIMAL SEX NUMBER M/F 31 M 32 M 33 M 34 M 35 M 36 M 37 M 38 M 39 M 40 M 41 M 42 M 43 M 44 M 45 M 46 M 47 M 48 M 49 M 50 M 51 M 52 M 53 M 54 M 55 M 56 M 57 M 58 M 59 M 60 M DEFINED AND FINAL STATE OF NECROPSY K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 TEST DAYS FIRST AND LAST DAY UNDER TEST DATE OF NECROPSY 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 CXR0481s Final Report Page 90 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 47/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 31 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 32 CXR0481s Final Report Page 91 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 48/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 33 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): -mononuclear cell infiltrate(s), unilateral, grade 1 * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 34 CXR0481s Final Report Page 92 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 49/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 35 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 36 CXR0481s Final Report Page 93 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 50/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 37 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 38 * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 39 CXR0481s Final Report Page 94 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 51/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE CONT./FF. ANIMAL NO. 39 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 40 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 41 CXR0481s Final Report Page 95 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 52/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 42 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 43 CXR0481s Final Report Page 96 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 53/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 44 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 45 CXR0481s Final Report Page 97 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 54/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 46 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): -mononuclear cell infiltrate(s), unilateral, grade 1 47 CXR0481s Final Report Page 98 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 55/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 48 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): -mononuclear cell infiltrate(s), unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present Organ examined, no pathologic findings noted 49 CXR0481s Final Report Page 99 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 56/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 50 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 51 CXR0481s Final Report Page 100 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 57/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 52 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 53 CXR0481s Final Report Page 101 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 58/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 54 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 55 CXR0481s Final Report Page 102 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 59/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 56 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral* * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 57 CXR0481s Final Report Page 103 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 60/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 58 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 **ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 59 * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 60 CXR0481s Final Report Page 104 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 02, PFOS 20 ppm PAGE CXR 61/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE CONT./FF. ANIMAL NO. 60 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 105 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. ANIMAL HEADING DATA DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 62/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT -07 PathDataSystem V6. 2c2 ANIMAL SEX NUMBER M/F 61 M 62 M 63 M 64 M 65 M 66 M 67 M 68 M 69 M 70 M 71 M 72 M 73 M 74 M 75 M 76 M 77 M 78 M 79 M 80 M 81 M 82 M 83 M 84 M 85 M 86 M 87 M 88 M 89 M 90 M DEFINED AND FINAL STATE OF NECROPSY K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 TEST DAYS FIRST AND LAST DAY UNDER TEST DATE OF NECROPSY 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 CXR0481s Final Report Page 106 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 63/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 61 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 62 CXR0481s Final Report Page 107 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 64/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 63 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 64 CXR0481s Final Report Page 108 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 65/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 65 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 66 CXR0481s Final Report Page 109 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 66/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 67 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 68 CXR0481s Final Report Page 110 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 67/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 69 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 **ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 70 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 71 CXR0481s Final Report Page 111 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 68/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE CONT./FF. ANIMAL NO. : 71 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* 72 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 73 CXR0481s Final Report Page 112 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 69/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 74 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 75 CXR0481s Final Report Page 113 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 70/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 76 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 1 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 77 CXR0481s Final Report Page 114 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 71/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 78 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 79 CXR0481s Final Report Page 115 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 72/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 80 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -fatty change (vacuolation), midzonal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 81 CXR0481s Final Report Page 116 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 73/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 82 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -fatty change (vacuolation), midzonal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 83 CXR0481s Final Report Page 117 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 74/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 84 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 85 CXR0481s Final Report Page 118 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 75/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 86 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 **ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 87 CXR0481s Final Report Page 119 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 76/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 88 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 89 CXR0481s Final Report Page 120 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 03, PFOS 100 ppm PAGE CXR 77/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 90 * MICROSCOPIC FINDINGS LIVER: -fatty change (vacuolation), midzonal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 CXR0481s Final Report Page 121 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. ANIMAL HEADING DATA DOSE GROUP : 04, PB 500 ppm PAGE CXR 78/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 ANIMAL SEX NUMBER M/F 91 M 92 M 93 M 94 M 95 M 96 M 97 M 98 M 99 M 100 M 101 M 102 M 103 M 104 M 105 M 106 M 107 M 108 M 109 M 110 M 111 M 112 M 113 M 114 M 115 M 116 M 117 M 118 M 119 M 120 M DEFINED AND FINAL STATE OF NECROPSY K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 TEST DAYS FIRST AND LAST DAY UNDER TEST DATE OF NECROPSY 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 CXR0481s Final Report Page 122 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 79/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 91 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 92 CXR0481s Final Report Page 123 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 80/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 93 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present Organ examined, no pathologic findings noted 94 CXR0481s Final Report Page 124 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 81/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 95 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 96 CXR0481s Final Report Page 125 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 82/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 97 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 -cyst, unilateral, grade 1 98 CXR0481s Final Report Page 126 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 83/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 99 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * MICROSCOPIC FINDINGS No microscopic findings noted. * ANIMAL NO. : 100 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 101 CXR0481s Final Report Page 127 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 84/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 102 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 103 CXR0481s Final Report Page 128 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 85/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 104 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 105 CXR0481s Final Report Page 129 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 86/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 106 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 107 CXR0481s Final Report Page 130 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 87/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 108 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 3 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Only one of paired organs examined/present -branchiogenic cyst, unilateral, grade 1 109 CXR0481s Final Report Page 131 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 88/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 110 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, centrilobular, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 111 CXR0481s Final Report Page 132 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 89/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 112 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 113 CXR0481s Final Report Page 133 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 90/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 114 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 115 CXR0481s Final Report Page 134 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 91/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 116 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 117 CXR0481s Final Report Page 135 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 92/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 118 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 2 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 119 CXR0481s Final Report Page 136 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 04, PB 500 ppm PAGE CXR 93/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 120 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 3 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 CXR0481s Final Report Page 137 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. ANIMAL HEADING DATA DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 94/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 ANIMAL SEX NUMBER M/F 121 M 122 M 123 M 124 M 125 M 126 M 127 M 128 M 129 M 130 M 131 M 132 M 133 M 134 M 135 M 136 M 137 M 138 M 139 M 140 M 141 M 142 M 143 M 144 M 145 M 146 M 147 M 148 M 149 M 150 M DEFINED AND FINAL STATE OF NECROPSY K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K1 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K2 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 K3 TEST DAYS FIRST AND LAST DAY UNDER TEST DATE OF NECROPSY 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 1 07-NOV-06 07-NOV-06 08-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 7 07-NOV-06 13-NOV-06 14-NOV-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 28 07-NOV-06 04-DEC-06 05-DEC-06 CXR0481s Final Report Page 138 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 95/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 121 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 122 CXR0481s Final Report Page 139 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 96/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 123 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 124 CXR0481s Final Report Page 140 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 97/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 125 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 126 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 141 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 98/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 127 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): -mononuclear cell infiltrate(s), unilateral, grade 1 * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 128 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -hypertrophy, hepatocellular, centrilobular, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 142 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 99/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 129 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): -ectopic thymus, unilateral * STATE AT NECROPSY: K1 DAYS ON TEST :1 * ANIMAL NO. : 130 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 143 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 100/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 131 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 132 CXR0481s Final Report Page 144 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 101/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 133 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 134 CXR0481s Final Report Page 145 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 102/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 135 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 136 CXR0481s Final Report Page 146 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 103/ 109 0481/s PATHOL. NO. 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : 137 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 * STATE AT NECROPSY: K2 DAYS ON TEST :7 **ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 138 * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 139 CXR0481s Final Report Page 147 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 104/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE CONT./FF. ANIMAL NO. : 139 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K2 DAYS ON TEST :7 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted* 140 * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 141 CXR0481s Final Report Page 148 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 105/ 109 0481/s PATHOL. NO. 07041 VOO DATE 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 142 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): -branchiogenic cyst, unilateral, grade 1 143 CXR0481s Final Report Page 149 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 106/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 144 * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 145 CXR0481s Final Report Page 150 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 107/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 146 * MICROSCOPIC FINDINGS LIVER: -mononuclear cell infiltrate(s), periportal/peribiliar, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 147 CXR0481s Final Report Page 151 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 108/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 148 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : * MICROSCOPIC FINDINGS LIVER: -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 3 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted 149 CXR0481s Final Report Page 152 of 153 PATHOLOGY REPORT INDIVIDUAL ANIMAL DATA TEST ITEM TEST SYSTEM SPONSOR : Perfluorooctane Sulfonate (PFOS) : RAT, 28 days, feeding : CXR Biosciences Ltd. TEXT OF MICROSCOPIC FINDINGS DOSE GROUP : 05, Wy-14,634 50 ppm PAGE CXR 109/ 109 0481/s PATHOL. NO.: 07041 VOO DATE : 12-OCT-07 PathDataSystem V6.2c2 MALE * STATE AT NECROPSY: K3 DAYS ON TEST : 28 * ANIMAL NO. : 150 * MICROSCOPIC FINDINGS LIVER: -(mixed) inflammatory cell infiltration, focal, grade 1 -vacuolation, hepatocellular, periportal, grade 1 -hypertrophy, hepatocellular, diffuse, grade 2 THYROID GLAND (BOTH LOBES): Organ examined, no pathologic findings noted CXR0481s Final Report Page 153 of 153