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REPORT Twenty-eight-day Toxicity Study Repeated of Sample Dose Oral D-1 in Rats (BMR 14 3 C) Bio-Medical Research Laboratories Co. L t d. i 91997 ,''Itil REPORT Twenty-eight-daY Toxicity Study Repeated of Sample Dose Oral D-1 in Rats (BMR 14 3 C) -TRANSLATION- Pur* free rife February 16, 1993 Bio-Medical Research Laboratories C o. , L t d. PREFACE BY TRANSLATOR This is a total translation of the original report in Japanese language. On this work. I have paid close attention to transfer the real facts and the correct meaning In the text, confirming scientific matters to the original auther, the study director Mr. Michlo Otsuka. April 23, 1997 Translator Makoto Tennichi General Manager Blo-Medical Research Laboratories Co., Ltd. CONTENTS STATEMENT ON GLP CERTIFICATION SIGNATURE GENERAL MATTERS Title / Study Number Purpose GLP Applicalton Testing Guidelines Sponsor / Testing Facilities / Responsible Person Dates In the Study Retention of Records and Samples SUMMARY_ MATERIALS AND METHODS 1. Test Substance 2. Preparation of the Dosing Solution 3. Laboratory Animals and Reason for Choice 4. Animal Housing 5. Identification of Animals 6. Identification of Cages 7. Test Procedures 7-1 Dose levels and reason for selection 7-2 Number of animals 7-3 Route, frequency, and duration of administration 7-4 Method of administration 7-5 Grouping (I) 8. Parameters assessed 8-1 General condition Clinical signs / Body weight / Food consumption 8-2 Laboratory tests Hematology / Biochemistry 8-3 Pathological examinaiton Necropsy / Organ weight 8-4 Histological examination 9. Statistical Analysis Urinalysis RESULTS1. Mortality 2. Clinical Signs 3. Body Weight 4. Food Consumption 5. Hematological Findings 6. Biochemical Findings 7. Urinalysis 8. Pathological Findings 8-1 Organ weight 8-2 Macroscopic findings at necropsy B-3 Histological findings DISCUSSION AND CONCLUSION FIGURE AND TABLES (Group mean data) Figure 1. Changes In mean body weight Table 1. Group Incidence of clinical signs Table 2. Body weight Table 3. Food consumption Table 4. Hematology (in connection with RBC) Table 5. Hematology (in connection with WBC) Table 6. Clinical chemistry Table 7. Urinalysis Table B. Organ weight (Absolute) Table 9. Organ weight (Relative) Table 10. Group incidence of macroscopic findings Table 11. Group Incidence of microscopic findings APPENDICES (Individualdata) Appendix 1. Clinical signs Appendix 2. Body weight Appendix Appendix Appendix 3. Food consumption 4. Hematology (in connection with RBC) 5. Hematology (in connection with WBC) Appendix 6. Clinical chemistry Appendix Appendix 7. Urinalysis 8. Organ weight (Absolute) Appendix 9. Organ weight (Relative) Appendix 10. Macroscopic findings Appendix 11. Microscopic findings Appendix 12. Microscopic findings (for the organs showed gross lesions) PHOTOGRAPHS (Histological findings) Photos 1-4 STATEMENT ON GLP Title Twenty-eight-day Repeated Dose Oral Toxicity Study of Sample D-1 in Rats Study Number BMR143C The said study has been conducted In compliance with the "GLP Standard for Industrial Chemicals" In Japan (1984, amended 1988). (Signature) (Date) Signed February 16, 1993 (Makoto Tennichl) General Manager Bio-Medical Research Laboratories Co., Ltd. (iv) CERTIFICATION Title Twenty-eight-day Repeated Dose Oral Toxicity Study of Sample D-1 In Rats Study Number BMR143C Having audited the final report, I, the undersigned, assure that the study has been conducted in compliance with Japanese "GLP Standard for Industrial Chemicals". and accomplished with the protocol and appropriate standard operating procedures of the testing facility, and the report has been prepared exactly based on the experimental results. (Signature) (Date) Signed February 16, 1993 (Yutaka Kambara) Quality Assurance Unit Bio-Medical Research Laboratories Co., Ltd. Dates of Inspection Object Dates of Inspection Dates of Report to Study Director to Management Protocol 1992. 6.23 1992. 6.23 1992. 6.23 1992. 7. 3 1992. 7. 3 1992. 7. 3 Testing Procedures 1992. 7. 7 1992. 8. 4 1992. 7. 7 1992. 8. 4 1992. 7. 7 1992. 8. 4 1992. 8.18 1992. 8.20 1992. 8.20 Records, Raw Data 1992.12.17 1992.12.17 1992.12.17 Final Report 1993. 2.16 1993. 2.16 1993. 2.16 (v) SIGNATURE Title: Twenty-eight-day Repeated Dose Oral Toxicity Study of Sample D-1 In Rats Study No.: BMR143C The said study has been conducted under the responsibility of the undersigned. and this report has been prepared so as to reflect whole study correctly. (Signature) (Date) Signed February 16, 1993 (Michlo Otsuka) Study Director Bio-Medical Research Laboratories Co., Ltd. (vi) GENERAL MATTERS 1. Title Twenty-eight-day repeated dose oral toxicity study of Sample D-1 in rats 2. Study Number BMR143C 3. Purpose To investigate toxicological potential of the test substance, Sample D-1. through serial oral administration to rats for 28 days. It was Intended to meet Japanese legal regulation for industrial new chemical substances. 4. GLP Application The study was conducted In compliance with the Japanese "GLP Standard for Industrial Chemicals" (1984, ammended 1988). 5. Testing Guidelines The testing methods conformed to the Japanese "Guidelines for Screening Toxicity Testings of Chemicals" (1988). 6. Sponsor Sumitomo 3M Co., Ltd. 3-8-8. Minami-hashimoto, Sagamihara, Kanagawa, Japan (Responcible Person) Nobushige Murakami Manager, Technical Department, Chemicals Division - 1- 7. Testing Facilities Key Facility (DBlo-Medical Research Laboratories Co., Ltd. 3079 Susugaya, Kiyokawa-mura, Aikoh-gun, Kanagawa, Japan Partial Contributor (Blood Biochemical Examination) Kashima Laboratory, Mitsubishi-Kasel Institute of Toxicological and Environmental Sciences 14, Sunayama, Hasaki-machl, Kashima-gun, lbaraki, Japan *)The company name has altered to Mitsubishi Chemical Safety Institute. (Histopathological Specimens Preparation) (3) Nara Pathological Research Kinomine. Hayama, Tuge-mura, Yamabe-gun, Nara, Japan (Histopathological Examination) Hatano Research Institute, Food and Drug Safety Center 729-5 Ochlai. Hadano-city, Kanagawa, Japan B..Responslble Personnel (Circled figure after the name indicates each belonging facility cited above.,) General Manager Makoto Tennichi, Study Director General Participants Michlo Otsuka, (D Hiroyuki Ishii, Naoto Ichikawa. Osamu Tanaka, Mikiko Kawanabe, Quality Assurance Yutaka Kambara, 2 Partial Participants (Responsible to biochemical determination) Naoto Toyota, (Responsible to histopathological specimens reparation) Yoshimi Tatsuni, (Director for histopathological examination) Shinsuke Yoshimura. (Participant to histopathological examination) Fumie Kawashima, 9. Dates in the Study (Initiationof the study) June 23, 1992 (Animals reception) July 1, 1992 (Administrationperiod) -Male- July 6 - Aug. 2, 1992 .-Female- July 7 - Aug. 3, 1992 (Termination of observation, blood sampling and necropsy) -Male, main groups- Aug. 3, 1992 -Female, main groups- Aug. 4, 1992 -Male, recovery groups- Aug. 17, 1992 -Female, recovery groups- Aug. 18, 1992 (Completion of the study) Feb. 16, 1993- 10. Retention of Records and Samples All documentation records and samples (including specimens) of the study are retained in the archives of Bio-Medical Research Laboratories Co.. Ltd. for 10 years after the completion of the study. As to further retention, it will be decided on consultation with the sponsor and the contractor. 3 SUMMARY 1 Sample D-1 was administered to both sexes of rats with doses of 0.1, 1, 10, and 30 mg/kg for 28 days. A group of rats given the vehicle (olive oil) alone was included as the control. Besides the main groups for nonrecovery study, a satellite group for 14-day recovery study was added to each of the highest dose and the control groups. 2. No death occurred throughout the administration period. One male of the 30 mg/kg group, however, died during the recovery period; the death was considered to be caused by the test substance. 3. No abnormal clinical signs were found In all rats of the control (including those for the recovery study) and the 10 mg/kg or lower dose groups throughout the whole of the observation period. In the 30 mg/kg group, however. such signs as the followings were observed during the administration and the recovery periods: reduced spontaneous movement, tonicclonic convulsion, lying or crouching, bradypnea or cyanosis, salivation, reddish salivation, brown urine, loss of hair, and coat staining. These were considered to be caused by the test substance. 4. Body weight gain was suppressed during the administration period in both males and females of the 30 mg/kg group from the second week and after; the suppression continued throughout the recovery period. 5. Food consumption was suppressed during the administration period in both males and females of the 30 mg/kg group from the second week and after; It tended to return during the recovery period, however. 6. Hematological examination at the end of the administration period revealed - 4- decreased hemoglobin concentration in males of the 10 and 30 mg/kg groups, and reduced prothrombin time in females given 30 mg/kg. Moreover. at the end of the recovery period, the followings were still detected in the 30 mg/kg group: decrease of erythrocyte count and hematocrit, shortening of prothrombin time, and increase of leukocyte count, In males; decrease of hemoglobin concentration and mean corpuscular hemoglobin concentration, shortening of activated partial thromboplastin time, and increase of mean corpuscular volume, In females. 7. Biochemical examination revealed the the followings at the end of the administration period: decrease of GOT in females of the I mg/kg and higher dose groups; increase of albumin In females of the 10 mg/kg and higher dose groups; increase of GPT and chloride, and decrease of total cholesterol In both sexes, decrease of total protein and increase of alkaline phosphatase and of A/G ratio In males, increase of urea nitrogen in females, all these in the 30 mg/kg group. At the end of the recovery period, the followings were observed In the 30 mg/kg group: decrease of triglyceride and increase of A/G ratio, In males; increase of urea nitrogen, total protein. albumin, and calcium, and decrease of glucose and triglycerides, in females. Besides, reddish change of serum was recognized in both sexes with the 10 mg/kg or higher dose main groups, and In the 30 mg/kg recovery group. 8. -By urinalysis. acidic shift in both sexes and decrease of protein in females were noted in the 30 mg/kg group during the administration period. Acidic shift remained In males during the recovery period. 9. As for absolute organ weight, the administration resulted In increase of the liver weight in both sexes of the 10 mg/kg or higher dose groups and decrease in the kidneys weight of males given 30 mg/kg. After the recovery 5 period, Increase in the liver of both sexes and decrease in the kidneys of females were still noted In the 30 mg/kg group. Relative organ weight increased In the liver of both sexes of the 10 mg/kg or higher dose and increased in the kidneys in females of the 10 mg/kg or higher dose. After the recovery period, increase in the liver of both sexes and increase In the kidneys of females remained In the 30 mg/kg group. 10. Autopsy for the male died during the recovery period revealed atrophy and dark reddish patch In the thymus, hemorrhage and tarry contents in the stomach, tarry contents in the small intestine, and grayish patch in the liver. In the scheduled necropsy at the end of the administration period, the following findings were detected by macroscopic examination: dark reddish change in the liver of females receiving 30 mg/kg; grayish dot or patch and hypertrophy In the liver and blackish change In the kidneys in males given 30 mg/kg; -dark reddish change, yellowish change, grayish patch, and hypertrophy In the liver of females receiving 30 mg/kg. The examination at the, end of the recovery period revealed grayish patch, dark reddish change, and hypertrophy In the liver of both sexes of the 30 mg/kg dose. 11. Histopathological examination at the end of the administration period revealed dose-dependent changes as eosinophilic degeneration in centrilobular hepatocytes and swelling in the liver In males of the 1 mg/kg or higher doses and In females of the 10 mg/kg or higher dose. Other findings: focal necrosis in hepatocytes in both sexes given 30 mg/kg and in males given 10 mg/kg; somewhat remarkable fatty degeneration In peripheral lobule in males receiving 30 mg/kg. No abnormal change was detected in the heart, kidneys, spleen, adrenals, brain, and testes or ovaries. 6 At the end of the recovery period, eosinophilic degeneration In centri:lobular hepatocytes turned slighter; swelling in the liver, however, did not recover in both sexes given 30 mg/kg, and focal necrosis also remained. In case of one male given 30 mg/kg and died during the recovery period, In addition to the same changes as found liithe liver of the main groups, the followings were detected: hemorrhagic foci in the brain, ulcer and hemorrhage In the glandular stomach, and congestion and hemorrhage in the thymus. 12. Judging from the above, the toxicological no-observed-effect level (NOEL) of the test substance, Sample D-1, was considered to be 0.1 mg/kg for both sale and female rats. MTERIALS AND METHODS 1. Test Substance 1-1 Name (Chemical Name) 2-[N-Ethyl-N-perfluoroalkyl(C-1-8)sulfonylamino] ethyl acrylate (Abbreviation) Sample D-1 1-2 Structural formula C2 H5 0 Cn F2n+lSO2 N-CH2 CH2 O-C.-CH=CH 2 (n ranges from 1 to 8) (Composition) about 78% for n= 8 component, about 21% for n=1-7 components. 1-3 Molecular weight 625 1-4 Lot number 101 (suppliedby the sponsor) 1-5 Appearance Amber-colored waxy solid., 1-6 Purity 99% or above 1-7 Strage conditions Stored at room temperature .2. Preparationof Dosing Solution 2-1 Vehicle Olive oil 2-2 Procedure Required amount of the test substance was weighed and suspended homogeneously in the vehicle mentioned above. The dosing solutionwas divided into daily batches and stored in a refrigerator.at 4 OC until just before use. 2-3 Frequency of preparation The dosing solution was prepared once a week, because the stability of the test substance in the mixture for eight days was confirmed In Japan Food Research Laboratories (Tokyo) by 8 our request. 3. Laboratory Animals and Reason for Choice The animals used were SPF Crj:CD(SD) rats of both sexes. They were purchased from Charles River Japan, Inc. (795 Shimo-furusawa, Atsugi, Kanagawa, Japan) at four weeks of age on July 1. 1992. After five (male) or six (female) days of acclimatization, animals that apparently healthy and showed normal weight gain were selected for the study at five weeks of age. They were grouped to be uniform for the mean body weight based on the last weighing during the acclimatization. The weight ranged from 132 to 151 g for males and 121 to 141 g for females at the Initiation of administration. Reason for choice: Having appropriate nature for laboratory animals and being bred under good genetic and microbial control. this species and strain are widely used in toxicological studies; our testing facility. besides, has accumulated back ground data on this animal. 4. Animal Housing Five or two (during or after the acclimatization,respectively) animals of the same sex were housed together in a stainless steel wire mesh cage of the hanging type (26X38Xl8 cm). The animal care room was a barriered facility tha t was automatically controlled to keep the following conditions: Temperature 22 - 26 -C Humidity 30 - 70% RH Ventilation 13 times/hour (all fresh air) Lighting Light /dark cycle of 12 hours (lights on 6:00 to 18:00) A little deviation of temperature and humidity occurred during the animal keeping; it was, however, judged not to affect to the reliability of the study. The diet was MF pelleted food for laboratory animals (Oriental Yeast Co., 9 Ltd.) and drinking water was tap water subjected to filtration and UV irradiation. Animals were allowed access to food and,water ad libitum except for fasting from the evening before necropsy. Analysis of the ingredients and microbial contamination of the diet was performed by the manufacturer, and toxic contaminants were examined by Japan Food Research Laboratories (Tokyo) at the manufacturer's responsibility.The quality of the tap water was periodically examined by Atsugi Public Health Center by our request. These data were checked and stored by us. Cages were replaced with freshly outoclaved ones once a week. 5. Identification of Animals Each animal was Identified by marking a number on the tail with a saturated alcoholic solution of picric acid. To identify the dose group to which each animal belonged, a colored mark was painted on the tail with an oil-based felt-tip pen. 6. Identification of Cages Each cage was identified with an attached card on which the study number, abbreviated name of the test substance, dosage, and Individual animal number were listed. 7. Test Procedures The testing methods conformed to the Japanese "Guidelines for Screening Toxicity Testing of Chemicals * (amended on Decenber 5, 1986). 7-1 Dose levels and reason for selection Prior to the principal study. preliminary dose-finding studies ranged between 4 and 14 days in administration period was performed with such doses as 20. 40, 50, 100, 200, 500, and 1000 mg/kg, each In a few (three to five) male and female rats. It resulted death of more than half number of rats in 10 1000 mg/kg for 14 days repeated administration; suppression of body weight gain was observed In 40 mg/kg dose, though none of changes found in 20 mg/kg dose. Hence, the highest dose was decided at 30 mg/kg in the principal study and the lower doses were set at 10 mg/kg, 1 mg/kg. and 0.1 mg/kg for both sexes. A group of rats given the vehicle alone was included as the control. 7-2 Number of animals Forty-two rats of each sex were used. 7-3 Route, frequency, and duration of administration Administration was performed via the oral route, once a day for 28 days. 7-4 Method of administration The dosing mixture was intubated directly Into the stomach via a plastic gavage tube. The volume administered was I ml per 100 g of body weight based on the most recent weight data. 7-5 Grouping Four dose groups mentioned above and the control group were set. Besides, to monitor the reversibility of any toxic effects, satellite groups were added to the control and the highest dose groups for a 14-day recovery study. Grouping by dose levels Dose (mg/kg) Main Groups(*i)- T Recovery Groups(*2) (Sex and Humber of animals) Control 0 male & female, 6 rats each male & female, 6 rats each Lowest dose 0.1 male & female. 6 rats each Intermediate-1 1 male & female, 6 rats each Intermediate-2 10 male & female, 6 rats each Highest dose 30 male & female, 6 rats each male & female, 6 rats each Total male & female,30 rats each male & female.12 rats each - 11 - (*I) Animals were necropsied on the next day of the last administratrion. (*2) Animals were necropsied Immediately after the recovery period was over. Note. Individual animal number. Each animal was numbered with four-digit figure. First digit, Dose groups; 0. the control group 11 the lowest dose group 2, the intemedlate-1 group 3. the lntemediate-2 group .4. the highest dose group Second digit, Sexes; 11 males 2. females Third and fourth digits, individual animal of main or recovery groups; 01-06, animals of the main groups 07-12, animals of the recovery groups 8. Parameters Assessed 8-1 General condition 8-1-1 Clinical signs The appearance, behavior, and excrete of every animal were observed daily throughout the administration and the recovery periods. 8-1-2 Body weight Every animal was weighed with an electric balance at the commencement of the administration, once a week thereafter. 8-1-3 Food consumption Leftover food from each cage was weighed on day 0 and once a week thereafter, and the daily average consumption per animal was calculated for each group. 8-2 Laboratory tests 8-2-1 Hematology Blood samples were collected from the inferior vena cava under ether anesthesia just before the scheduled time of sacrifice. Blood sampling was 12 done under non-fasting condition. The hematological parameters determined are listed below. The anticoagulant used was ethylenediaminetetraacetic acid dipotassium salt (EDTA-2K), except that 3.13% sodium citrate was used to determine the prothrombin time and the activated partial thromboplastine time. Parameters (1) Leukocyte count (2)Erythrocyte count (3)Platelet count (4)Hemoglobin concentration (5)Hematocrit (6)Mean corpuscular volume (7)Mean corpuscular hemoglobin (8)Mean corpuscular hemoglobin concentration (9)Differential leukocyte count (10)Prothrombin time (11)Activated partial thrombo- plastin time (WBC) (RBC) (PLT) (Hgb) (Hct) (MCV) (MCH) (MCHC) (PT) (APTT) Method Electrical resistance method Electrical resistance method Electrical resistance method Photo-electric colorimetry Pulse detection method Calculated from (2) and (5) Calculatedfrom (2) and (4) Calculatedfrom (4) and (5) Giemsa-stained smear Quick's one-stage test Activated cefaroplastin method Insturuments (I)-(B) Automatic cell counter, Sysmex model CC-180A, TOA Medical Electronics Co.,Ltd. (9) Optical microscopy (10)-(11) Blood coagulater, Model KC-1. Amelung Go.,Ltd. Among these data, PT and APTT of one male in the recovery group (animal no. 0111) were missing owing to coagulation at blood sampling. On the other hand, reticulocyte counting was added in the main groups after sequential staining with new methylene blue process and May-GruenwaldGlemsa process, because anemia was suspected in animals of this group. 8-2-2 Biochemistry The residual blood sample was let stand at room temperature for 30 min and - 13 - then centrifuged at 3000 rpm for 10 min to separate serum. The serum, being frozen to -20 OC, was sent to Kashima Laboratory of Mitsubishi-Kasel Institute of Toxicological and Environmental Sciences for biochemical examination. Parameters examined were as follows: Parameters Method (1) Total protein Bluret's method (2)Albumin Bromocresol green method (3)A/G ratio Calculated from (1) and (2) (4)Glucose Enzymatic UV-spectrophotometry (Hexokinase-G6-PDH) (5)Triglycerides Enzymatic method (LPL-GK-G3PO-POD) (6)Total cholesterol Enzymatic method (CES-CO-POD) (7)Urea nitrogen Enzymatic UV-spectrophotometry (Urease-GLDH) (8)Creatinine Jaffe method (9)Calcium o-Cresolphthalein complexone method (10) Inorganic phosphorous Phosphomolybdate-UV spectrophotometry (11)GOT (AST) UV spectrophotometry-rate method (Modified SSCC) (12)GPT (ALT) UV spectrophotometry-rate method'(Modified SSCC) (13) 7,-GPT 7,-Glutamyl-p-nitroanilide substrate method (Modified SSCC-) (14)Alkaline phosphatase (ALP) p-Nitrophenyl phosphate substrate method (15)Sodium Ion selective electrodes method (16)Potassium Ion selective electrodes method (17) Chloride Ion selective electrodes method Instrument: Automatic analyser (Hitachi 736-10 type) 8-2-3 Urinalysis Fresh urine was collected from all animals at six or seven days before the completion of administration, and four or five days before the end of the recovery period. (In each of two successive days, the former and the latter days were for male and for female respectively.) These were tested for the following parameters. 14 Parameter pH, Occult blood, Protein, Glucose, Ketone bodies, Urobilinogen, Bilirubin B-3 Pathological examination Method Paper test, MULTISTIX (9) (Miles-Sankyo Co.,Ltd.) 8-3-1 Becropsy and macroscopic examination On the next day of the last administration, all survived animals except those assigned for the recovery test were sacrificed for necropsy after blood sampling; whereas for animals of the recovery groups, the necropsy was held on the scheduled day just after the recovery period was over. After careful Investigation for any macroscopic abnormalities, the following organs were removed and fixed in 10% neutral buffered formalin: brain, pituitary, eye ball, (with Harderian glands), thyloid (with parathyloid), heart, lungs, liver, kidneys, spleen, adrenals, stomach, testes or ovaries, urinary bladder, and right femur (with bone marrow). 8-3-2 Organ weight The following organs were weighed before fixation, then the relative organ weight (ratio of organ weight to body weight on the day of necropsy) was calculated: brain, liver, kidneys, adrenals, and testes or ovaries. 8-4 Histological examination From the following fixed organs of all rats in the control and the highest dose (30 mg/kg) groups, histological sections of paraffin-embeded specimens were cut and subjected to hematoxylin-eosin staining, then histological changes were surveyed: heart, liver, spleen. kidneys, adrenal$, brain, and testes or ovaries. (typical seven organs) The survey of the liver was expanded to animals of the other dose groups since some chages were found in case of 30 mg/kg dose group. 15 Additional histological survey was conducted to the following cases in which any macroscopic changes were observed at the necropsy: lung in one case (animal no. 0105), kidneys in two cases (animal nos. 1103, 2102). and testes in one case (animal no. 0111). Concerning the dead animal which occurred during the recovery period in one sale (animal no. 4109), histological survey was performed In the typical seven organs and in thymus and stomach, the latter two were additional because of the macroscopic changes found at the necropsy. In this animal, tarry intestinal contents were found between duodenum and ileum at autopsy; however, preparation of the specimen and histological examination were not performed because of post-mortem autolysis All through the histopathology, histological specimens were prepared in Nara Pathological Research, whereas the microscopic examination was carried out in Hatano Research Institute of Food and Drug Research Center, both at our quest. 9. Statistical Analysis For metrical data, homogeneity of variance among all groups was first tested by Bartlett's method. Then the homogeneity of all group means was tested by one-way layout analysis or Kruskal-Wallis H-test, depending on whether the group variance was homogeneous or not, respectively. When group means were significantly heterogeneous and each group had the same number of data items, differences in mean values between any group and the control were tested by Dunnett's multiple range test or Dunnett's rank sum test, according to whether the group variance was homogeneous or not, respectively. When group means were heterogeneous and the number of data items In each group was unequal, the above-mentioned procedures were replaced by Scheffe's multiple range test or Scheffe's rank sum test, respectively. Armitage's chi-squared test was used to assess discrete data such as urinalysis findings. 16 RESULTS 1. Mortality All rats survived to the end of administration period; during the recovery period, however, one male given 30 mg/kg died on day 32 (viz. 'day 4 of recovery *). Each day throughout the study is indicated with such way as the day of commencement of the administration is named "day 0 and the next day.of the last administration (viz. day 28), that is the first day of recovery period, is expressed as "day 0 of recovery 2. Clinical Signs (Table 1. and Appendix 1) 2-1 During the administration period No abnormal signs were noted in all fats of the control and the 10 mg/kg or lower dose groups. With rats receiving 30 mg/@g (inclusive of the recovery group). however, the following findings were detected: Salivation---a considerable number of animalt of both sexes, In the latter half of administration period; and In one male on day 0, besides; Reddish salivation---in a few males and females. occasionally; Tonic-clonic convulsion, lying, bradypnea---in one male on day 26: Reduced spontaneous movement, clonic. or tonic-clonic convulsion, lying or crouching, bradypnea or cyanosis---in one and two females on day 20 and from day 25 to 27, respectively; Loss of hair and coat staining---In three and one females respectively, on day 17 and after. 2-2 During the recovery period No abnormal signs appeared in both males and females of the control group as well as in the administration period. In case of the male given 30 17 mg/kg and died during the recovery period, reduced spontaneous movement_ lying, bradypnea, reddish salivation and brownish urine were found on day 31 (viz. day 3 of recovery). then died on the next day. Loss of hair found in'females of the 30 mg/kg group during the administration period lasted until the termination of the recovery period, whereas the coat staining in one female disappeared after day 34 (viz. day 6 of recovery). 3. Body Weight (Fig.1, Table 2, Appendix 2) 3-1 During the administration period Body weight gain In the 10 mg/kg or lower dose groups was similar to that of the control throughout the administration period. Significant suppression of weight gain, however, appeared in both males and females receiving 30 mg/kg (rats of the recovery group inclusive) from week 2 and after. 3-2 During the recovery period The above-mentioned suppression of weight gain during the administration period lasted until the end of recovery period. 4. Food Consumption (Table 3. Apendix 3) 4-1 During the administration period Food consumption in the 10 mg/kg or lower dose groups was similar to that of the control throughout the administration period. Significant suppression of the consumption, however, was observed in both sexes receiving 30 mg/kg (rats of the recovery group inclusive) from week 2 and after. 4-2 During the recovery period The suppression in females of the 30 mg/kg group lasted one more week after the finish of the administration. 18 5. Hematological Findings (Table 4 and 5, Appendix 4 and 5) 5-1 At the end of the administration period Hematological examination with animals of the main groups performed on the scheduled necropsy-day revealed the following changes: decreased hemoglobin concentration in males of the 10 mg/kg and higher dose groups; reduced prothrombin time in females receiving 30 mg/kg. 5-2 At the end of the recovery period After-recovery examination with animals of the 30 mg/kg recovery group revealed the following changes: in males, decrease of erythrocyte count, hemoglobin concentration, and hematocrit, shortening of prothrombin time. and increase of leukocyte count; in females, decrease of hemoglobin concentration and mean corpuscular hemoglobin concentration, shortening of activated partial thromboplastin time-, and increase of mean corpuscular volume. 6. Biochemical Findings (Table 6. Appendix 6) 6-1 At the end of the administration period The examination with animals of the main grotfps resulted in decrease of GOT in females of the I mg/kg and higher dose group, and increase of albumin in also females of the 10 mg/kg and higher dose groups. Moreover, in the 30 mg/kg group, the following changes were significant: increase of GPT and of chloride, and decrease of total cholesterol, all these In both sexes; decrease of total protein and Increase of alkaline phosphatase and of A/G ratio, all these in males; and- increase of urea nitrogen in females. In addition, reddish change of serum was visually recognized in two males (animal nos. 3102, 3103) and In one female (animal no. 3205) given 10 mg/kg. and in five males (animal nos. 4101, 4102, 4103, 4105, 4106) and In all females given 30 mg/kg. 19 6-2 At the end of the recovery period Significant changes detected in the 30 mg/kg recovery group were as follows: lowering of triglycerides and increase of A/G ratio In males; increase of urea nitrogen, total protein. albumin, calcium, inorganic phosphorous and sodium, and decrease of glucose and triglycerides, all these in females. And besides, reddish change of serum was recognized in four males (animal nos. 4108, 4110, 4111, 4112) and in three females (animal nos. 4207, 4210, 4211). 7. Urinalysis (Table 7. Appendix 7) 7-1 During the administration period The examination performed with rats (inclusive of the recovery group) one week before the end of administration period resulted in the following changes: pH shift to alkaline side in male'sof the 1 mg/kg and 10 mg/kg groups and In females receiving 0.1 mg/kg; pH shift to acidic side in both sexes receiving 30 mg/kg; decrease of occult blood in males receiving 30 mg/kg; decrease of protein in females receiving 30 mg/kg. 7-2 During the recovery period Shift of pH to acidic side was noted in males of the 30 mg/kg recovery group In the examination conducted one week before the end of the period. 8. Pathological Findings 8-1 Organ weight (Table 8 and 9; Appendix 8 and 9) B-1-1 Absolute organ weight 1) At the end of the administration period With the animals of the main groups, liver weight increased In both males 20 and females of the 10 mg/kg and higher dose groups; whereas weight decreased in kidneys of males and ovaries of females, both in the 30 mg/kg group. 2) At the end of the recovery period Changes noted in the 30 mg/kg recovery group were as follows: increase of the liver weight in both sexes; Increase in the testes of males; decrease In the kidneys, In the adrenals and in the ovaries, all these of females. 8-1-2 Relative organ weight 1) At the end of the administration period Increase of liver weight found in the absolute weight with the 10 mg/kg and higher dose groups was also detected in the relative weight in both males and females. In addition, the followings were observed: increase in the brain of both sexes receiving 30 -mg/kg; increase in the adrenals and the testes of mal.esreceiving 30 mg/kg: increase in the kidneys of females given 10 mg/kg and higher dose. 2) At the end of the recovery period Changes detected in the 30 mg/kg recovery group were as follows: increase in the liver and the brain of both sexes; increase in the testes of males; increase in the kidneys of females. 8-2 Macroscopic findings at necropsy (Table 10; Appendix 10 and 12) 8-2-1 Macroscopic findings in the dead animal Autopsy on the male died during the recovery period (animal no. 4109) revealed the following changes: atrophy and dark reddish patch In the thymus; hemorrhage and tarry contents in the stomach; tarry contents in the small intestine; grayish patch in the liver. 8-2-2 Macroscopic findings at scheduled necropsy - 21 - Gross pathological findings observed were listed below. 1) At the end of the administration period Males of the 30 mg/kg group---greyish dot or patch in the liver and hypertrophy of the liver. in all six rats; blackish change In the kidneys and dark reddish change in the adronals. in one rat. Females of the 30 mg/kg group---dark reddish change, yellowish change and grayish patch in the liver, in four, one and one rats respectively; hypertrophy of the liver, in all six rats; pyelectasis In the kidneys of one rat. Females of the 10 mg/kg group---dark reddish change In the liver in one rat. Others---graysh patch In the kidneys of one male of the 1 mg/kg group; pyelectasis in the kidneys of one male of the 0.1 mg/kg group; cyst in the kidneys and dark reddish patch in the- lung, each in one male of the control group. 2) At the end of the recovery period Males of the 30 mg/kg group---grayish patch and dark reddish change, and hypertrophy in the liver, all these In two rats. Females of the 30 mg/kg group---dark reddish change and grayish patch, and hypertrophy in the liver, in five, one, and one rat respectively. Others---atrophy of the testes In one mala of the control. 8-3 Histological findings (Table 11; Appendix 11 and 12) 8-3-1 Histological findings in the dead animal Histopathological examination with the male died during the recovery period (animal no.4109) revealed the following findings: eosinophilic degeneration in centrilobular hepatocytes, focal necrosis and peripheral fatty change in the liver; dilated tubules in restricted part of the 22 - kidneys; hemorrhagic foci in the brain; ulcer and hemorrhage in theglandular stomach; congestion and hemorrhage in the thymus. No abnormality was obvserved In the heart, spleen. adrenals and testes. 8-3-2 Histological findings at scheduled necropsy Histological findings observed were listed below. 1) At the end of the administration period Liver In centrilot5ular hepatocytes, cytoplasm appeared to be granular and eosinophilic, and swelling was recognized in all males and females of the 30 mg/kg group; In the 10 mg/kg group, the similar changes, though slightly, were found In all males; and in females, either the swelling or eosinophilic change was observed in four rats even slighter In incidence and intensity.than in male; in the 1 mg /kg group, the similar change was slighter in intensity and incidence than in 10 mg/kg grcup. (Photo I and 2). Focal necrosis in hepatocytes was found in five males and two females of the 30 mg/kg group, and in two males of the 10 mg/kg group. The necrosis was found also in one male of the control. Fatty change in peripheral lobule was remarkable in males of the 30 mg/kg group (Photo 4); In females, however, the incidence and intensity was similar to that In the control. Localized fatty change was also observed in one male of the control. Microglanuloma was found in a few males and females of all groups, though clear difference in Incidence and intensity was not noticed between each dose group and the control group. (2)Heart No abnormalities were found except extremely localized myocardinal degeneration in two male of the control group. 23 (1)Spleen No abnormalities were found except extramedullary hematopolesis In three male of the control group. (3) Kidneys Regenerated tubules were found in both males and females of the 30 mg/kg and the control groups though no difference in incidence and intensity between two groups was noted. Other findings: dilated tubules in one Male of the control; Infiltration of lymphocytes in the interstitial tissue in one male of the 30 mg/kg group and three male and female of the control; dilation of pelvis In two males and in one female of the 30 mg/kg group. Adrenals, brain and testes or ovaries No abnormalities were observed. 2) At the end of the recovery period Liver Centrilcxbular hepatocytes were eosinophilic and showed swelling in all males and females given 30 mg/kg. As compared to the main group animals, eosinophillc intensity turned slighter In both sexes; however, the intensity of swelling was similar to that in the main group. Focal necrosis In hepatocytes was found in three males and two females of the 30 mg/kg group, though none In the control. Other findings: fatty change in peripheral lobule and microglanuloma were observed, the former was somewhat definite in males given 30 mg/kg, the latter, however, no difference was found with the control. Heart Very slight myocardial fibrosis was found in one male of each of the 30 mg/kg and the control group, and myocardial degeneration was found in one female of the cotrol. No other abnormal findings were noted. (3) Spleen No abnormal findings were found except extramedullary hematopolesis in one 24 male of each of the 30 mg/kg and control groups. (a)Kidneys Regenerated tubules were found In both sexes of every group; infiltration of lymphocytes in Interstitial tissue was observerd in one male and in two female of the 30 mg/kg and the control group, respectively; very slight chronic nephropathy was found in one male given 30 mg/kg. No other abnornmalities was found. Testes No abnormal findings were noticed except atrophy in one male of the control group. Adrenals, brain and ovaries No abnormalities were observed. 3) Histological findings for gross pathological abnormalities Swelling of cytoplasm of centrilobular hepatocytes and focal necrosis were found corresponding to the macroscopic findings of hypertrophy and grayish patch, respectively. 25 DISCUSSION AND CONCLUSION Major findings obtained through repeated oral administration of the test substance to rats for 28 days were summarized below. Those changes were considered to be affected by the test substance. m: male, f: female Period Administration Recovery Examination Findings Dose \ (mg/kg) 0.1 1 10 30 30 Clinical signs (*l)---reduced spontaneous movement, bradypnea, anemia, salivation, convulsion; sup- m pression of body weight gain and food consump- f f tion ............................................................................................................................... death occurred (number of the animal) m (1) Hematological findings---Inclination to anemia, mm m abnormal change of blood coagulation f f Biochemical findings---Abnormal changes in lipids, m m protein, and in enzymes or in electolytes f ff f Urinalysis (*2)---acidicurine m m f Organ weight ---change in absolute weight of the mm m liver; ff f .................................................................................................................................................... change in relative weight of the liver and kid- m m ...m....... neys ff f Macroscopic findings---greyish dot or patch and m m hypertrophy In the liver f f Histological findings---such changes as eosinophil- mmm m ic swelling and focal necrosis in the liver, ff f fatty *c**h.*.a.n..g..e..i..n. peripher.a..l..l..o..b.ule m (*l)Observation during each of the administration and the recovery period. (*2)Examination one week before each end of the administration and the recovery period. The other- examination was practised at each end of the administration and the recovery period. 26 [Changes suggesting relation with the liver] Among the changes detected, decrease of serum cholesterol was remarkable. Though the mechanism of the reaction is not clear, It is generally accepted that cholesterol is produced through synthesis in the liver and absorption of lipid in the intestine and is controlled through accumulation in tissues, conversion into energy and excretion from the body. As decrease of total protein and increase of A/G ratio in males or increase of albumin In females were found, considering the connection among protein and lipoprotein and cholesterol, influence of the test substance on globulin or effect to decrease lipoprotein was suggested; hence, for the reduction of serum cholesterol, concern of the liver was suspected. Lowering of triglyceride found in both males and females of the 30 mg/kg recovery group was also considered to- be affected by the test substance; because the same tendency was observed In the main group though statistically insignificant and the facts that the test substance was suspected to affect to lipids as mentioned above. Moreover, changes In enzymes relating to the liver, increase of the liver weight such macroscopic findings in the necropsy as grayish dot/patch and hypertropby, and histopathological pictures such as eosinophilic swelling and*focal necrosis in centrilobular hepatocytes which were considered to be connected with the macroscopic findings, all these changes. suggested that the test substance affected to the liver parenchyma. Additional changes suggesting relation with the liver were shortened prothrombin time in females of the 30 mg/kg main group and in males of the same dose of the recovery group, shortened activated partial thromboplastine time In females of the 30 mg/kg recovery group. Changes of calcium and glucose observed in females of the 30 mg/kg recovery group, though not found in the main group, were a little remarkable comparing with level of physiological variation, and connection with the administ- 27 ration of the test substance was presumed. [Changes suggesting relation with other organs] As' to the Increase of leukocyte count observed In the 30 mg/kg recovery group, the mechanism was unclear; the same tendency, however, was -noticed in the main group though statistically insignificant. Therefore the connection with the test substance was suspected. For the increase of urea nitrogen found In females given 30 mg/kg in both the main and the recovery groups, it was not clear.whether the change was caused by degradation of. protein or by excretion disturbance in the kidneys; however, considering such facts as the increase of chloride in both sexes of the 30 mg/kg main group and the increase of the kidneys weight in females of the 10 mg/kg or higher dose main groups and of the 30 mg/kg recovery group, effect of the test substance to the kidneys was suspected, though no histological changes were found. As to reddish change of serum recognized visually in both sexes of the 10 and 30 mg/kg groups, sometimes -Y-GTP activity does not change apparently when reddish serum was caused by hemolysis. In this case of the study, however. the relation among the reddish serum and hemolysis and 7,-GTP was unclear. [Changes considered-not to be related to the test substance] The following changes were considered not to be related to the effect of the test substance. Changes of Inorganic phosphorous and sodium observed in females of the 30 mg/kg recovery group were unconnected to the test substance, because the magnitude of the changes was within limit of physiological variation and the changes were not found in the main group. In urinalysis, significant decrease of occult blood was recorded in males of the 30 mg/kg main group; it was, however, considered to be apparent, 28 because one case of occult blood occurred in the control accidentally, hence the change was detected statistically. Urine pH shifted to alkaline side with the administration in males receiving 1 and 10 mg/kg and in females receiving 0.1 mg/kg; the change, however, was not dose-dependent, and the change was considered not to be related to the test substance, therefore. Weight increase of the brain and the testes and decrease of the ovaries were observed in males or females given 30 mg/kg in both the main and the recovery groups; no histological findings, however, were detected. Therefore, the changes were considered to be apparent caused by decrease of the body weight and to have no toxicological significance. As for the macroscopic changes in the adrenals found in males or females given 30 mg/kg, no histological findings were detected; they were Judged to have little toxicological significance,therefore. Concerning any other changes such as gross pathological findings having no dose dependency and no histologocal findings except above-mentioned incidents, they were Judged to be unconnected with the test substance. [Conclusion] As 'mentioned above, significant response was found in the liver by administering the test substance to both sexes of rats with 1 mg/kg or higher dose. Whereas none of response was observed in the 0.1 mg/kg group with both male and female. Therefore the toxicological no-observed-effect level (NOEL) of the test substance, Sample D-1. was concluded to be 0.1 mg/kg for both male and female of rats. 29 .v F I G.URES AND TA8LES (Group Mean Data) 9 400- B 250- 0 d v w 250- 20me t 150 f Study No. BMR143C Mal$ x Administration 1'4 i Recovery 2@ 4@O DaA after oommorcownt 6f treated 9 B 250- 0 d y 3m- w 2cA - h 200- t 150 Fem 19 ------N-'--' Administration 7' 1'4 2'1 i Recovery 1 1 28 35 Days after wmmencoment of treated Fig. 1 Changes in mean body weight of male and female rats during 28 days of oral administration of Sample D-1 and 14 days recovery period. 13 0 mvkg + 0.1 mg/kg I uwkg & 10 mg/kg X 30 mg/kg Table 1-1 Group incidenceof clinicalsigns Testarticle Sample D-1 Sex lale Dose level Sex Clinicalsigns (mg/kg) Days after commencement/ cessationof treatment 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 /0 29 30 31 32 3 0 No abnormalityobserved 12 ......................................................................................................................>..1..2......6.................................... 0.1 No abnormalityobserved 16 ........................................................................................>.6................. 1 No abwraelity observed 6 .......................-.........................................................................................................>..6................................... 10 No abnormalityobserved 6 ............................................................................................>..6............................................... 30 Reduced spontaneous movement Tonic-clonic/Clonicconvulsion Male Lying Crouching Salivation 1 Reddishsalivation 2 22 2 55 5 3 2 3 4 3 Reddish snivel Bradypnea Cyanosis Brownishurine Loss of hair Coat staining No abnormalityobserved 11 12 ..........................................................>...1.2..1.0 10 9 10 7 7 7 9 10 9 7 8 12 6 6 6 5 5 Death 1 Sex Fenile Table I-F Group incidenceof clinicalsigns Test article Sample D-1 Sex Fewle st Dose level Clinicalsigns (mg/kg) Days aftercomencesent / cessationof treatment 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27.i /i,,29 30 3132 33 3 0 lo abnormalityobserved -12 ............................................:...................................................................................... 0.1 No abnormalityohserved 6 .............................-....................................................................................................@..1.2..-.....:..6.................................... 1 0a lit obs-TrvW-6 ............................................................................................................... 10 30 meduced spontaneous sovement . .........................................................................................................................>..6................ Tonic-clonic/Clonicconvulsion 21 L2 Lying 2 121 Crouching Salivation Reddishsalivation Reddish snivel 2 2 1 2 23 4 5 22 1 Bradypnea cyawsis Brownishurine Lossof hair Coat staining No abnormalityobserved Des th 3 I ............................................................................. 12 .........................................>.1.1..1.1..1.1...8....6...6...5...5...4....7...5...7...7...4 8 2 2 2 2 2 2 Dose level 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean S.D. N Mean S.D. N Mean S.D. N Mean S.D. N Mean S.D. N Table 2m Body weight - Group mean values Sex Male Test article Sample D-1 Study No Unit g Weeks after commencement/ cessation of treatment 0 1 2 3 4 5/1 143 204 260 5.6 8.8 12.2 12 12 12 143 207 269 4.6 6.6 16.7 6 6 6 144 206 265 4.2 7.1 9.9 6 6 6 143 201 257 5.8 8.8 14.2 6 6 6 143 193 239 3.7 7.0 10.2 12' 12 12 icaiitldyifferent from control value 4 307 15.4 12 320 24.7 6 314 13.2 6 305 19.6 6 272 14.5 12 P<0.05 342 21.8 12 361 26.8 6 359 31.8 6 349 17.7 6. 343 24.5 6 290 16.2 12 315 t 13.4 5 P<o.ol. Table 2 -F Body weight - Group mean values Sex Female Test article Sample D-1 Study No Unit : g Bose level Weeks after commencement/ cessation of treatment 0 1 2 3 4 5/1 0 mg/kg 0.1 mg/kg I mg/kg 10 mg/kg 30 mg/kg tlean 130 S.D. 5.4 N 12 166 193 217 237 8.1 13.1 16.5 18.6 12 12 12 12 Mean 129 164 189 212 227 S.D. 3.4 2.3 5.0 9.6 10.5 N 6 6 6 6 6 Mean S.D. N 131 167 192 217 235 6.6 13.7 14.2 15.2 15.9 6 6 6 6 6 Mean 130 161 185 203 214 S.D. 5.9 7.9 9.8 13.4 14.4 N 6 6 6 6 6 Mean 130 158 171 176 185 S.D. 3.9 7.0 7.2 7.8 10.9 N 12 12 12 12 12 Significantlydifferent from contro-l---va-Fu-e-, P<0.01. 5 264. 20.7 6 195 9.9 6 Table 3 -M Food consumption - Group mean values Sex Male Test article Sample D-1 Study Unit Dose level Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean S.D. N 16.8, 0.85 6 19.1 0.92 6 18.9 0.77 6 Mean S.D. N 18.0 1.26 3 20.6 1.88 3 20.2 2.96 3 @leaii S.D. N 17.0 0.75 3 19.3 0.57 3 18.1 1.00 3 Mean 16.6 18.3 19.5 S.D. 1.25 1.61 2.77 N 3 3 .3 Mean 15.4 15.8 tt 15.3 t S.D. 0.72 0.68 0.36 N 6 6 6 Significantlydifferent from control value 6 16.8 1.27 6 18.8 2.5. 2 3 21.8 1.26 3 23.0 0.75 3 17.3 0.65 3 17.8 2.63 3 12.7 0.64 6 P<F.-05 20.4 2.46 3 21.2 2.12 3 P<U.ul. Table 3F Food consumption Sex : Test article : Group mean values Female Sample D-1 Study No. Unit Dose level Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean S.D. N 13.8 1.15 6 13.1 1.36 6 13.9 1.50 6, Mean S.D. N 13.7 0.40 3 12.6 0.80 3 13.4 1.42 3 Mean 14.0 12.3 12.9 S.D. 2.35 1.55 1.07 N 3 3 3 Mean S.D. N 12.7 1.10 3 11.5 0.81 3 11.2 0.75 3 Mean S.D. N 12.9 0.78 6 10.3 0.72 .6 8.1 0.82 6 Significaiitlv-'@ifferefnrtom control value 7 12.7 1.42 6 12.4 1.12 3 11.9 0.79 3 9.7 0.36 3 8.0 1.09 6 P<0.05 19.4 0.20 3 20.0 0.35 3 14.3 t 1.51 3 15.4 1.97 3 P<0.01. Table 4 -M Hematology - Group mean values Sex Male Testarticle Sample D-1 Animalskilledon scliedul(e4 weeks) Dose level RBC HB Ht count conc. (XI04/MMI) (g/di) % mcv (,am3' MCH (ps) 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean 735 14.2 47.3 S.D. 46.0 0.39 2.66 .N 6 6 6 Mean 698 14.0 46.7 S.D. 33.7 0.18 2.74 N 6 6 6 Mean 14.0 47.2 S.D. 38.8 0.43 3.21 N 6 6 6 Mean 728 13.4 t 46.2 S.D. 31.4 0.33 2.15 .N 6 6 6 Mean 700 13.3 t' 46.1 S.D. 53.3 0.85 3.30 N 6 6 6 Significantldyifferentfrom controlvalue 65 19.3 2.9 1.02 6 6 67 20'.1 1.0 0.91 6 6 64 19.0 1.9 0.69 6 6 64 18.4 2.4 0.80 6 6 66 19.1 1.5 1.01 6 6 p<0.05. MCHC Plate'let PT count % (xiol-lme) (sec) 30.0 96.6 14.3 1.66 12.88 0.59 6 6 6 30.1 93.8 14.0 1.54. 10-51 0.44 6 6 6 20.7 103.7 14.0 1.34 18-92 0.31 6 6 6 28.9 103.7 13.9 1.20 11-79 0.30 6 6 6 29.0 89.9 14.4 1.63 6.76 0.29 6 6 6 Table 4M 2 Hematology - Group mean values Sex Male Test article Sample D-1 Animalskilledon schedule(Recovery) Dose levei RBC HB Ht count conc (XI04/MMI) (P,/jd) % mcv (,U M3 MCH (pg) MCIIC % Platelet count (xlo4/MMI) PT (sec) 0 mg/kg 30 mg/kg Mean 798 14.1 45.5 S.D. 54.4 0.39 2.82 N 6 6 6 Mean 717 12.7 41-.6 S.D. 22.6 0.50 1.68 N 5 5 5 Significantldyifferentfrom controlvalue 57 17.7 31.0 2.2 .0-88 1.44 6 6 6 58 17.7 30.4 2.2 0.67 1.15 5 5 5 p<0.05 p<0.01. 88.4 16-90 6. 95.5 11.28 5 13.8 0.11 5 13.4 0.47 5 9 Table 4 -F I Hematology - Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(4 weeks) Dose level RBC HB Ht mcv MCH MCHC count coric. (xIC@/miP (g/dl) % (AM3) (pg) % 0 mg/kg Mean 705 13.3 42.8 S.D. 51.2 0.43 3.36 N 6 6 6 61 18.9 31.3 2.5 1.00 1.70 6 6 6 0.1 mg/kg Mean 691 13.6 42.0 S.D. 25.2 0.18 1.40 N 6 6 6 61 19.7 32.4 3.4 0.66 1.24 6 6 6 1 mg/kg Mean 701 13.3 41.3 S.D. 52.0 0.29 1.79 N 6 6 6 60 19.4 32.2 2.3 1.09 1.19 6 6 6 10 mg/kg Mean 716 13.7 43.8 S.D. 35.6 0.30 2.06 N 6 .6 6 61 19.2 31.4 2.7 0.96 1.00 6 6 6 30 mg/kg Mean 722 13.0 43.1 S.D. 54.6 0.49 2.95 N 6 6 6 60 18.0 30.1 1.5 1.16 1.92 6 6 6 Significantldyifferentfromcontrolvalue 10 Platelet count (XI04/MM3) PT (sec) 115-7. 14.3 15-64 0.20 6 6 103.1 14.3 12-27 0.46 6 6 107.9 14.2 6.81 0.57 6 6 105.9 14.0 13.81 0.42 6 6 103.2 13.4 11.39 0.21 6 6 Dose level 0 mg/kg 30 mg/kg Table 4 F-2 Hematology - Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(Recovery) RBC HB nt mcv MCH MCHC count conc. (xl(@/mm3) (s/di) % (IIM3) (pg) % Mean 715 13.7 43.1 S.D. 24.6 0.28 1.68 N 6 6 6 Mean 678 12.6 42 7 S.D. 33.3 0.29 1.;2 N 6 6' 6 Si&iiificantdlfyffereiiftromcontrolvalue 60 19.2 31.8 2.3 0.38 1.12 6 6 6 63 18.7 29.6 1.3 0.61 0.80 6 6 6 p<5.0-5 p<0.01. Platelet count (XIVI=3) PT (sec) 93.8 13.5 13-44 0.37 6 6 110.3 13.1 17.87 0.46 6 6 -- Table 5 -m -I Hematology - Group mean values Sex Male Test article. Sample D-1 Animals killed on schedule (4 weeks) Study No Dose level WBC Differentialcount of leukocytes % of total counte count Lympho- Neutrophils Eosino- Baso- (Xio'/mm') cytes Segmented Band phils phils 0 mg/kg Mean 88 86 8' 1 0 0 S.D. 22.6 5.7 3.2 0.5 0.5 0.0 N 6 6 @6 6 6 6 0.1 mg/kg Mean 106 87 8 0 1 0 S.D. 24.8 5.7 3.9 0.5 1.2 0.0 N 6 6 6 6 6 6 1 mg/kg Mean 118, 88 8 0 1 0 S.D. 23.3 4.3 3.1 0.4 0.5 0.0 N 6 6 6 6 6 6 10 mg/kg @lean 97 85 10 0 1 0 S.D. 17.4 5.6 3.5 0.0 1.0 0.0 N 6 6 6 6 6 6 30 mg/kg Mean 137 86 9 1 0 0 S.D. 65.5 2.8 2.4 0.8 0.0 0-o N 6 6 6 6 6 6 $ : Statisticalanalysis impossible. 12 Dose level 0 mg/kg 30 mg/kg Table 5 -M -2 llematolo&y- Group mean values Sex Male Test article Sample D-1 Animals killed on schedule (Recovery) Study No WBC Differentialcount of leukocytes of total counte count Lympho- Neutrophils Eosino- Baso- (xlO'/mm') cytes Segmented Band phils phils Mean 97- 85 it 0 0 0 S.D. 13.9 6.5 6.1 0.5 0.0 0.0 N 6 6 6 6 6 6 Mean 126 87 8 1 0 0 S.D. 11.6 4.6 1.8 0.5 0.0 0.0 N 5 5 5 5 5 5 : Significantli-di-ffe-rferioimt control value $ : Statisticalanalysis impossible. p<0.01. 13 Table 5 F-I Hematology Group mean values Sex Female Test article Sample D-1 Animals killed on schedule (4 weeks) Study No Dose level WBC count (x 102/mm' Differentialcount of leukocytes % of total counte Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 0 mg/kg Mean 75 86 10 1 0 0 S.D. 10.8 3.0 2.1 0.8 0.0 0.0 N 6 6 6 6 6 6 0.1 mg/kg Mean 101 88 7 0 0 0 S.D. 30.7 4.7 4.0 0.5 0.4 0.0 N 6 6 6 6 6 6 I mg/kg Mean 86 88 to 0 0 0 S.D. 21.9 4.4 3.7 0.5 0.5 0.0 N 6 6 6 6 6 6 10 mg/kg Mean 85 89 5 0 0 0 S.D. 22.3 3.1 2.1 0.4 0.5 0.0 N 6 6 6 6 6 6 30-mg/kg Mean 77 87 8 0 0 0 S.D. 15.6 4.4 4.8 0.4 0.4 0.0 N 6 6 6 6 6 6 $ Statisticalanalysis impossible. 14 Table 5 -F -2 Hematology Group mean values Sex Female Test article Sample D-1 Animals killed on schedule (Recovery) Study No. Dose level WBC Differentialcount of leukocytes % of total counted count Lympho- Neutrophils Eosino- Baso- (xlO'/mm') cytes Segmented Band phils phils 0 mg/kg Mean 85 86 9 0 1 0 S.D. 23.9 6.3 5.3 0.4 1.3 0.0 N 6 6 6 6 6 6 .30 mg/kg Mean 86 91 5 0 0 0 S.D. 18.1 2.3 2.2 0.4 0.4 0.0 N 6 6 6 6 6 6 $ Statisticalanalysis impossible. 15 Table 6-M -I Stu Clinicalchemistry- Group mean values Sex Male Test article Sample D-1 Animalskilledon schedule(4 weeks) Bose level GOT GPT R-GTP ALP Urea Creatinin Glucose Tot nitrogen cho (IU/I) (IU/I) (IU/I) (IU/I) (mg/di) (mg/dl) (mg/di) (mg 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean 68 30 0 711 S.D. 7.0 4.6 0.0 129.3 N 6 6 6 6 Mean .65 30 0 677 S.D. 9.0 5.1 0.0 84.9 N 6 6 6 6 Mean 71 31 0 734 S.D. 8.1 -4.5 0.0 127.7 N 6 6 6 6 Mean 65 34 0 780 S.D. 3.9 3.2 0.0 130.2 N 6 6 6 6 Mean 73 53 0 1191 S.D. 9.5 13.2 0.0 243.7 N 6 6, 6 6 4 Significantldyifferentfrom controlvalue , ** 7 $ : Statisticalanalysisimpossible. 16 12.4 1.62 6 10.6 2.18 6 13.3 2.34 6 12.4 2.56 6 14.1 1.93 6 0.5 0.00 6 0.5 0.05 6 0.5 0.04 6 0.5 0.05 6 0.6 0.12 6 140 9.4- 6 6 150 10.5 3 6 140 6.9 4 6 142 9.0 11 6 142 11.5 2 6 Table 6 M -2 Clinicalchemistry- Group mean values Sex Male Test article Sample D-1 Animalskilledon schedule(4 weeks) Study No. BMRI Dose .level Total protein (g/dl) Albumin (8/di) A/G ratio Calcium Inorganic Na K c phos. (mg/dl) (mg/dl) (mEq/1) (mEq/1) (mE 0 mg/kg Mean 6.51 3.98 1.58 9.8 10.2 142 4.1 1 S.D. 0.212 0.125 0.110 0.46 0.70 0.5 0.08 2 N 6 6 6 6 6 6 6 0.1 mg/kg Mean 6.38 3.96 1.63 9.8 9.5 142 4.0 1 S.D. 0.166 0.103 0.076 0.28 0.98 0.4 0.08 0 N 6 6 6 6 6 6 6 1 mg/kg Mean 6.22 3.84 1.63 9.8 9.5 142 4.2 1 S.D. 0.190 0.079 0.149 0.31 0.72 0.8 0.20 1 N 6 6 6 6 6 6 6 10 mg/kg Mean 6.20 3.93 1.74 9.8 9.3 142 4.3 1 S.D. 0.099 0.146 0.141 0.30 0.82 0.9 0.33 1 N 6 6 6 6 6. 6 6 30 mg/kg Mean 6.01 4.01 2.04 9.4 10.0 144 4.2 1 S.D. 0.368 0.166 0.264 0.54 3.06 1.9 0.35 1 N 6 61 6 6 6 6 6 Significantldyifferentfrom controlvalue ; P<0.01. 17 Table 6M 3 st Clinicalchemistry Group mean values Sex Male - Test article Sample D-1 Animilskilledon schedule(Recovery) Bose level GOT GPT r -GTP ALP Urea Creatinin Glucose To nitrogen ch (IU/I) (IU/I) (IU/I) (IU/I) (mg/di) (mg/di) (mg/dl) (m 0 mg/kg tlean 85 34 0 613 18.7 0.5 148 S.D. 12.7 4.5 0.0 65.2 2.42 0.05 5.9 1 N 6 6 6 6 6 6 6 30 mg/kg Mean S.D. N 76 44 6.5 10.1 5 5 0 716 21.8 0.5 152 0.0 105i4 3.61 0.08 12.8 1 5 5 5 5 5 Sigiiiifcaiiy from controlvalue $ Statisticalanalysisimpossible. P<0.01. Table 6 - M -4 Clinicalchemistry- Group mean values Sex Male Test article Sample D-1 Animal-skilledon schedule(Recovery) Study No. BMR Bose level Total protein (g/di) Albumin (&/dl) A/G ratio Calcium (mg/dl) Inorganic Na phos. (m&/di) (mEq/1) K (mEq/1). (mf 0 mg/kg Mean 6.42 3.85 1.50 9.6 8.7 141 4.2 1 S.D. 0.167 0.056 0.072 0.28 0.18 1.5 0.28 1 N 6 6 6 6 6 6 6 30 mg/kg Mean 6.57 4.14 1.70 9.7 8.9 142 4.2 1 S.D. 0.288 0;195 0.109 0.08 0.63 1.1 0.20 N 5 5 5 5 5 5 5 t Significantldyifferentfromcontrolvalue p<0.05. 19 Table 6 -F-I Stu Clinicalchemistry- Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(4 weeks) Dose ,level GOT GPT r -GTP ALP Urea Creatinin Glucose Tot nitrogen cho (IU/I) (IU/I) (IU/I) (IU/I) (ms/di) (mg/dl) (m&/di) (mg 0 mg/kg Mean 80 23 0 472 11.9 0.5 133 S.D. 8.0 3.5 0.4 65.2 1.33 0.04 7.6 10 N 6 6 6 6 6 6 6 0.1 mg/kg Mean 70 25 0 419 13.2 0.6 140 S.D. 5.0 4.8 0.0 84.6 3.41 0.08 22.0 6 N 6 6 6 6 6 6 6 I mg/kg Mean 60 20 0 437 13.1 0.5- 143 S.D. 7.4 2.9 0.4 82.1 2.89 0.06 8.3 5 N 6 6 6 6 6 6 6 10 mg/kg Mean 60 24 0 451 13.5 0.5 135 S.D. 12.5 3.2 0.0 81.8 2.59 0.04 11.7 8 N 6 6 6 6 6 6 0 30 mg/kg Mean 56 36 0 590 18.0 0.5 126 S.D. 4.1 7.2 0.0 170.9 3.43 0.05 15.0 5 N 6 6 6 6 6 6 6 Significantldyifferentfrom controlvalue -*-'*.-p<0.61. 20 Table 6 - F -2 Clinicalchemistry- Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(4 weeks) Study No. BMR Dose ,level Total protein (8/di) Albumin (g/di) A/G ratio Calcium Inorganic Na K phos. (mg/di) (mg/di) (mEq/1) (mEq/1) (m 0 mg/kg Mean 6.63 4.18 1.72 9.4 8.4 S.D. 0.173 0.060 0.099 0.15 1.09 N 6 6 6 6 6 141 4.1 0.8 0.27 6 6 0.1 mg/kg Mean 6.61 4.19 1.74 9.8 9.7 141 4.5 1 S.D. 0.320 0.154 0.095 0.59 1.01 0.9 0.58 N 6 6 6 6 6 6 6 1 mg/kg Mean 6.45 4.15 1.82 9.6 8.7 S.D. 0.335 0.125 0.191 0.18 0.31 N 6 6 6 6 6 141 4.4 1.2 0.48 6 6 10 mg/kg Mean 6.91 4.42 1.77 9.7 7.9 S.D. 0.255 0.143 0.1072 0.35 0.73 N 6 6 6 6 6 142 4.1 1.5 0.45 6 6 30 mg/kg Mean 7.01 4.59 1.90 9.9 8.6 S.D. 0.327 0.140 0.143 0.49 0.93 N 6 6 6 6 6 142 4.9 1.1 0.12 6 6 Significantldyifferentfrom controlvalue P<0.05 21 Table 6F3 Stu Clinicalchemistry Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(Recovery) Dose level GOT (IU/I) GPT (IU/I) r -GTP (IU/I) ALP (IU/I) Urea Creatinin Glucose TO nitrogen ch( (mg/di) (mg/di) (mg/di) (mi 0 mg/kg 30 mVI(g Mean 71 30 0 323 17.3 S.D. 8.8 7.7 0.0 63.4 2-55 N 6 6 6 6 6 Mean 61 30 0 387 24.1 S.D. 10.6 5.9 0.0 52.4 2.99 N 6 6 6 6 6 : Significantldyifferentfromcontrol value $ : Statisticalanalysisimpossible. P<0.01. 0.6 0.05 6 0.6 0.04 6 150 6.9 1 6 126 9.6 6 22 Table 6-F -4 Clinicalchemistry- Group mean values Sex Female Test article Sample D-1 Animalskilledon schedule(Recovery) Study No. BMR14 Dose level Total protein (s/dl) Albumin (&/di) A/G ratio Calcium (ms/di) Inorganic Na phos. (mg/di) .(mEq/1) K (mEq/1) ci (mEq/ 0 mg/kg Mean 7.02 4.34 1.62 9.6 7.3 140 4.1 105 S.D. 0.286 0.142 0.064 0.16 -0.54 0.6 0.12 1.2 N 6 6 6 6 6 6 6 6 30 mglkg Mean 7.50 4.72 1.70 10.2 8.1 4 141 4.3 105 S.D. 0.379 0.204 0.123 0.19 0.31 0.4 0.39 1.0 N 6 6 6 6 6 6 6 6 t Significantldyifferentfromcontrolvalue P<0.05 P<0.01 23 Table 7 - M - 1 Urinalysis- Group mean values Sex Male Test article Sample D-1 3 weeks after commencementof treatment Study Dose level N Urobilinogen (EU/di) Occult Blood Bilirubin Ketone bodies (mg/dl)' Glucose (g/di) Protein (mg/di) (mg/ks) 0.1 1 2 4 ?=8 TR 1+ 2+ 3+ - 1+ 2+ 3+ 5 15 40 ?=80 0. 1 0.25 0.5 1 TR 30 100 9300 $ $ 0 12 12" 0 0 0 0 9 2 0 0 1 12 0 0 0 1 10 1 0 0 12 0 0 0 0 0 2 10 0 0 0.1 6 600 00 5 1 00 0 60 0 0 04 1 1 0 6 0 0 0 0 0 0 5 1 0 1 6 6 0 0 0 0 5 1 0 0 0 6 0 0 0 0 5 1 0 0 6 0 0 0 0 0 .0 4 2 0 10 66 0 000 60000 6 0 00 06 00 0 6 0 0 0 0 0 0 4 2 0 30 12 12 0 0 0 0 12 0 0 0 0 12 0 0 0 0 8 2 2 0 12 0 0 0 0 0 1 9 2 0 Negative, TR 4 Trace, 1+ : STi-gTit2,+ Moderate, 3+ Severe $ statisticaalnalysis impossible. Significantldyifferentfromcontrolvalue p<0.05 p<0.01. 24 Table .7 - M 2 Urinalysis- Group mean values Sex Male Test article Sample D-1 5 weeks after commencementof treatment(Recovery) Study Doselevel N Urobilinogen (EU/di) Occult Blood Billrubin Ketone bodies (mg/dl) Glucose Protein (ms/dl) (m&/ks) 0.1 1 2 4 ?=8 - TR 1+ 2+ 3+ - 1+ 2+ 3+ - 5 15 40 ?=80 - 0.1 0.250.5 91 - TR 30 100 ?=300 0 6 6 0 0 0 0 2 4 0 0 0 6 0 0 0 1 5 0 0 0 6. 0 0 0 0 0 2 4 0 0 30 6 5 0 0 0 0 400 10 50 0 0 05000 5 0 0 0 0 0 0 5 0 0 : Negative,TR Trace,1+ : Sliglit2,+ : Moderate,3+ : Severe $ : Statisticaalnalysis impossible. : Significantldyifferentfromcontrolvalue P<0.01. 25 Table 7 - F - I Urinalysis- Group mean values Sex Female Test article Sample D-1 3 weeks after commencement of treatment Study N Dose level N Urobilinosen (EU/di) Occult Blood - Bilirubin Ketone bodies (msldl) Glucose (8/di) Protein (mg/di) (mg/k&) 0.1 1 2 4 2@8 - TR 1+ 2+ 3+ - 1+ 2+ 3+ - 5 15 40 ?-:80- 0.1*0.250.5 @l - TR 30 100 ?='300 0 12 12' 0 0 0 0 10 2 0 0 0 12 0 0 0 9 3 0 0 0 12 0 0 0 0 0 2 10 0 0 0.1 660000 42 000 6 0 0 0 5 1 000 6 0 0 0 00 2 4 0 0 1 6600 0 0 2 22 00 6 0 0 0 24 0 0 0 6 0 0 0 00 1 5 0 0 10 660000 5 1 000 6000 33000 6 0 0 0 00 2 4 0 0 30 12 12 0 0 0 0 7 3 2 0 0 12 0 0 0 10 2 0 0 0 12 0 0 0 0 3 5 4 0 0 - : Negative,TR : Trace, 1+ : S I!glit,2+ : Moderate, 3+ Severe $ : Statisticalanalysis impossible. : Significantldyifferentfromcontrolvalue p<0.05 p<0.01. 26 Table 7 - F - 2 Urinalysis- Group mean values Sex Female Test article Sample D-1 5 weeks after commencement of treatment(Recovery) Study Dose level N Urobilinogen (EU/di) Occult Blood Bilirubin Ketone bodies (m&/di) Glucose (8/di) Protein (mg/di) (mg/kg) 0.1 1 2 4 9;8 - TR 1+ 2+ 3+ - 1+ 2+ 3+ - 5 15 40 80 - 0.1 0.250..5 91 - TR -30 1W 9 3W 0 6 6 0 0 0 0 3 2 0 10 6 0 0 0 6 0 0 0 0 6 0 0 0 0 1 3 2 0 0 30 6 6 0 0 0 0 2 4 0 0 0- 6 0 0 0 6 0 0 0 0 6 0 0 0 0 0 4 2 0 0 - : NegativeT,R Trace..1+ : Sliglit@2.+ Moderate, 3+ Severe $ Statisticaalnalysis impossible. 27 Table 8 m-I Organ weight (Absolute) - Group mean values Sex Male Test article Sample D-1 Animals killed on schedule (4 weeks) Study No. Dose level F.B.W (g) Brain (g) Liver (8) Kidneys (g) Adrenals (mg) 0 mg/kg 0.1 mg/kg I mg/kg 10 mg/kg 30 mg/kg Mean S.D. N 351 22.0 6 1.96 0.117 6 13.60 1.180 6 2.43 0.156 6 50.5 3.19 6 Mean S.D. N 359 31.8 6 1.98 0.094 6 14-50 1.194 6 2.55 0.257 6 54.3 4.90 6 Mean S.B. N 349 17.7 6 1.94 0.045 6 13-88 1.459 6 2.26 0.170 6 52.9 3.71 6 Mean S.D. N 343 24.5 6 1.94 0.078 6 16.93 1.179 6 2.39 0.101 6 49.5 5.14 6 Mean S.D. N 292 17.6 6 1.97 0..072 6 20.36 2.228 6 2.11 0.084 6 50.4 4.15 6 F.B.W Final body weight Significantlydifferentfrom control value 28 p<0.01. Testes (g) 2.99 0.265 6 3.14 0.196 6 3.14 0.194 6 3.01 0.,189 6 3.08 0.250 6 Table 8M 2 Organ weight (Absolute) Group mean values Sex Mai*e Test article Sample D-1 Animals killed on schedule (Recovery) Study No. Dose level F.B.W Brain Liver Kidneys Adrenals (mg) Testes 0 mg/kg 30 mg/kg Mean S.D. N 389 34.4 6 2 '.05 0.051 6 13.57 1.768 6 2.47 0.151 6 53.1 6.85 6 2.74 0.942 6 Mean S.D. N 341 18.5 5 2.02 0.058 5 20-66-** 1.693 5 2.40 0.157 5 51.5 7.31 5 3.38 0.117 5 F.B.W Final body weight :.Significantldyifferentfrom control value p<0.05 p<0.01. 29 Table 8 -F Organ weight (Absolute) - Group mean values Sex Female .Test article Sample D-1 Animals killed on schedule (4 weeks) Dose level F.B.W (&) Brain Liver Kidneys (g) Adrenals (mg) 0 mg/kg 0.1 mg/kg 1 mg/kg 10 mg/kg 30 mg/kg Mean S.D. N 230 17.4 6 1.85 0.067 6 8.76 0.471 6 Mean S.D. N 227 10.5 6 1.79 0.054 6 8.65 0.679 6 Mean S.D. .N 235 15.9 6 1.91 0.069 6 9.07 0.852 6 Mean S.D. N 214 14.4 6 1 .85 0.063 6 10-58 0.973 6 Mean S.D. N 190 1.83 13.78 9.5 0.064 0.816 6 6 6 F.B.W F-ii-ia-T-Yowdeyight Significantlydifferentfrom control value 30 1.62 0.177 6 1.61 0.162 6 1.67 0.081 6 1.70 0.109 6 1.50 0.126 6 60.1 5.46 6 59.8 6.93 6 62.6 7.79 6 60.8 8.01 6 55.6 6.28 6 p<0.01. Study No. Ovaries (mg) 83.7 10.13 6 83.7 7.86 6 92.0 15-11 6 75.8 8.53 62.6 7.57 6 Table 8 F -2 Organ weight (Absolute) Group mean values Sex Female Test article Sample D-1 Animals killed on schedule (Recovery) Study No. Dose level F.B.W Brain (g) Liver* (g) Kidneys Adrenals (mg) Ovaries (mg) 0 mg/kg 30 mg/kg Mean S.D. N 277 24.8 6 1.93 0.074 6 9. 09 1.122 6 Mean S.D. N 208 14.5 6 1.86 0.131 6 12-46 0.574 6 F.B.W Final body weight Significantlydifferentfrom control value 1.78 0.100 6 1.59 0.146 6 70.8 12-55 6 54.0 7.39 6 p<0.05 89.7 9.96 6 68.5 6.22 6 p<0.01. 31 Dose level F.B.W Table 9 m-1 Study No. Organ weight (Relative percentage of body weight) - Group mean values Sex Male Test article Sample.D-1 Animals killed on schedule (4 weeks) Brain Liver-- Kidneys Adrenals 'Testes (Xlo-,) 0 mg/kg 0.1 mg/kg 1 ms/kg. 10 mg/kg 30 mg/kg Mean S.D. N 351 22.0 6 0.56 0.037 6 3.88 0.197 6 Mean S.-D. N 359 31.8 6 0.55 0.066 6 4.04 0.210 6 Mean S.D. N 349 17.7 6 0.56 0.037 6 3.97 0.272 6 Mean S.D. N 343 24.5 6 0.57 0.043 6 4.98 0.611 6 Mean S.D. N 292.** 17.6 6 0.68 0.048 6 6.97 0.475 6 F.B.W Final body weight Significantlydifferentfrom control value 32 0.69 0.026 6 0.71 0.018 6 0.65 0.060 6 0.70 0.060 6 0.72 0.035 6 P<0.05 14.5 1.55 6 15.3 2.70 6 15.2 1.03 6 14.5 1.10 6 17.3 1.54 6 0.86 0.094 6 0.88 0.071 6 0.90 0.057 6 0.88 0.075 6' 1.06 0.112 6 P<0.01. Dose level F.B.W Table 9 -M -2 Study No. Organ weight (Relative percentage of body wei&ht) - Group mean values Sex Male Test article Sample 0-1 Animals killed on schedule (Recovery) Brain Liver- Kidneys Adrenals Testes (xlo--,) 0 mg/kg 30 mg/kg Mean S.D.. N 389 34.4 6 0.53 0.044 6 3.48 0.160 6 0.64 0.056 6 Mean S.D. N 341 18.5 5 0.59 0.030 5 6.06 0.482 5 0.70 0.029 5 F.B.W Final body weight Significantlydifferentfrom control value p<0.05 13.7 1.82 6 15.2 2.67 5 0.70 0.231 6 0.99 0.069 5 p<0.01. 33 Dose level F.B.W (8) Table 9FI Study No Organ weight (Relative percentage of body weight) - Group mean values Sex Female Test article Sample D-1 Animals killed on schedule (4 weeks) Brain Liver Kidneys Adrenals. Ovaries (XIO-3 (XIO-3) 0 mg/kg 0.1 mg/kg I mg/kg 10 m&/kg 30 mg/kg Mean S.D. N 230 0.80 3.81 0.70 17.4 0.037 0.198 0.043 6 6 6 6 26.2 2.76 6 Mean S.D. N 227 0.79 3.81 0.71 10.5 0.053 0.148 0.042 6 6 6 6 26.4 2.89 6 Mean S.D. N 235 15.9 6 0.81 0.047 6 3.85 0.237 6 0.71 0.043 6 26.6 2.73 6 Mean S.D. N 214 0.87 4.95 0.79 14.4 0.055 0.255 0.015 6 6 6 6 28.4 2.54 6 Mean S.D. N 190 0.96 7.24 0.79 9.5 0.034 0.321 0.053 6 6 6 6 29.2 2.50 6 F.B.W Final body weight Significantlydifferentfrom control value 34 p<0.01. 36.4 4.28 6 36.9 2.90 6 39.3 6.90 6 35.4 3.23 6 33.0 4.37 6 Dose level F.B.W (g) Table 9 -F -2 Study No. Organ weight (Relative percentage of body weight) - Group mean values Sex Female Test article Sample D-1 Animals killed on schedule (Recovery) Brain Liver Kidneys Adrenals (XIO-3) Ovaries (XIO-3) 0 iig/kg 30 mg/kg Mean S.D. N. 277 24.8 6 0.70 0.059 6 3.28 0.157 6 0.64 0.035 6 25.6 4.10 6 Mean S.D. N 208 tt 14.5 6 0.90 0.047 6 6.00 0.358 6 .0-76 -0.033 6 25.9 2.43 6 F.B.W Final body weight Significantlydifferentfrom control value *t p<0.01. 32.5 2.25 6 33.0 3.18 6 35 Organ Pindings Period Sex Dose level(mg/kg) Number of animals Thymus Dark reddish patch Atrophy Lungs Dark reddish patch Liver Scatterring of grayish dOL/patch Ilypertrophy Greyish patch Dark reddish change Yellowish change kidneys Cyst Pyelectasis Greyish patch Blackish change Adrenals Dark reddish change Stomach Tarry contents/Ilemorrhage Testes Atrophy Table TD-N.P Group Incidence of macroscopic findings Test article Sample D-1 29 days kale Female 0 o.i 1 10 so 0.1 1 to so 6 6 6 6 6 6 6 6 6 6 0 0 0 0 0 0 0 0 0 0 0 0 0 -0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 0 0 6 00 0 0 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 00 0 1 0 00 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0. 0 0 0 6 0 0 0 0 1 0' 0 0 0 4 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Study No Recover De Male Female 0 30 z----su 6 5 6 6 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 2' 0 2 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 1 0 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 36 Table ti-M Group Incidenceof microscopicfindings (28days) Sex Male Test article Sample D-L Dose level 0 mg/kg (n-B) 0.1 mg/kg (n-G) 1 mg/kg (n-G) Findings Grade :t + * 141 -*+ *m Liver microgranuloma focal necrosis focal fatty change peripheral fatty change swellingof centrilobular hepatocytes cosinophilic change In centrilobular hepatocy-tes lleart myocardial degeneration S:Yocardial fibrosis leen extramedullary hematopolesis Kidneys regenerated tubules dilated tubules Infiltrationof lymphocytes dilationof pelvis -chronic nephropathy Adronals abnormality Brain abnormality Testes atrophy 042 00 5 0 100 5 0 100 6 0 000 6 0 000 6 000 0 4 200 0 6 000 0 3 2 10 0 3 120 0 5 0 10 0 3 2 1- 0 0 6 00 00 6 00 00 6 00 00 6 00 00 6 00 00 0 60 00 6000 0 6 0 00 0 5 100 0 60 0 00 6 00 00 Negative. t Very slight.+ Slight. No4orate. lff Severe - + 4+ 04 2 00 6 00 00 6 00 00 4 20 00 330 0 0 24 00 0 10 mg/kg + (n-6) 4+ N 03 30 0 4 0 2 .0 0 60 0 0 0 22 2 0 0 0 15 0 0 01230 Study NO.BN 30 mg/k 03 10 60 12 00 00 60 60 60 31 60 51 40 60 60 60 60 tt7 Bose level Findings Grade Liver microgranulosa focal necrosis focal fatty change peripheral fatty change swellingof centrilobular hepatocytes eosinophille change In centrilobularhepatocytes lleart myocardial degeneration myocardial fibrosis Spleen extramedullary hematopolests Kidneys regenerated tubules dilated tubules infiltrationof lymphocytes dilationof pelvis chronic nephropalhy Adrenals abnormality Brain abnormality Ovaries abnormality Negative. Very sIIght.+ Table lt-F Group incidenceof microscopicfindings (2$days) . Sex Female Test article Sample B-1 0 mg/kg (a-G) + 4+ m 0.1 mg/kg (n-S) - + *m 1 mg/kg (a-g) - + 1 50 00 6 00 00 6 00 00 2 2200 0 5 1 00 6 00 00 60 00 0 024 0 0 6 0 0 00 6 00 00 6 000 0 6 0 00 6 0 00 0 6 0 00 0 60 0 00 3 2 1 0 .0 6 00 00 3 2 100 6 00 00 6 00 00 6 0 0 00 60 0 00 6 00 00 Si Lght. Moderate. M Severe 24 0 00 6 00 00 6 0 00 0 32 10 0 60 0 00 6 00 0 0 10mg/kg (n-6) + Study NO.BK 30 mg/k 05 1 00 6 0 0 @O 0 60 0 00 2 22 0 0 24 00 0 24 0 00 13 40 60 31 00 00 60 60 60 33 60 60 50 60 60 60 60 38 Sol Doselevel Findings Grade Liver microgranulosa focal necrosis focal fatty change peripheral ratty change swelling of contrilobular hePALOCYLes eosinophille change In centrilobular hepatocytes Ileart myocardialdegeneration myocardial fibrosis spleen extramedullary hematopolesls Kidneys regenerated tubules dilated tubules Infiltrationof lymphocytes dilation of pelvis chronic nephropathy Adronals abnormality Brain abnormality Testes atrophy Ovaries abnormality Negative. Very slight.+ Tableti-R-1 GroupIncidencoef microscopifcindings(Recovery) Testarticle SampleD-1 StudyN Male Female 0 mg/kg (n-B-F 30 @gTk-g--In---ST- --U-mg/kg (n-B) 30 mg/k + 4+ m + *m + + 132 00 600 00 6 0 0 0 .0 60 00 0 60 00 0 6000 0 60 00 0 5 100 0 5 1 00 0 3 300 0 60 00 0 6000 0 6 000 0 6 000 0 6 000 0 6 000 0 5 0010 0 1 4 00 203 00 500 00 2 1200 00 050 00 320 50 000 4 1000 4 1000 4 1 000 50 000 40 100 50 000 4 1000 50 000 50 000 50 000 Slight. 4+ :Moderate. M Severe 06 000 60 000 60 000 4 1 10 0 6 000 0 6000 0 5 100 0 6000 0 6 000 0 2 4 -0 0 0 6000 0 4 020 0 6000 0 6 000 0 6 000 0 6 000 0 60 00 0 1 32 402 6 00 501 0 02 0 06 6 00 6 00 6 00 510 6 00 6 00 6 00 6 00 6 00 6 00 6 00 Findings Sex Dose level Grade Liver Eosinophillechange In centrilobularbepatocytes Svelling or centrilobular hepatocytes Focal necrosis Peripheral fatty change Kidneys Dilated tubules Brain liamorrhagic foe[ Stomach Ulcer and hemorrhage In glandular stomach Thymus Congestion liesorrhage Ileart No abnormallty Spleen No abnormality Adrenals No abnormality Testes No abnormality Negative. Very slight.+ Table 11-R-2 Incidenceof microscopicfindingsIn died sale rat (32 day) Testarticle: SampleD-1 Kale 30 mg/kg (n-1) + 000 1 0 0 00 10 0 00 10 0 0 100 00 100 00 100 00 100 00 100 00 100 10 000 10 000 10 000 10 0 0 0 Slight. i+ Noderato. M Severe Study - - -W A P P E N D I C E S (Individual Data) Sex Dose level Animal (mg/kg) number 0101 0 4- 0112 1101 0.1 4 1106 2101 t Male 2106 3101 10 3106 4101 4102 4103 30 4104 4105 4106 4107 4108 4109 Appendix 1-1-1 Clinicalsigns- Individual Test article Sample D-1 Sex Vale Study Days after commencement 0 1*2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 -- - -- - -- - - -- - -- - -------- - -- - - -- -- - - -- - -- -------- - - - - - - - - - - - - --- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -- - - - - - - - - - - - - - - - - - - - - - - - SA SA SA SA -- - -- - -- - -- - - -- --------- - - - - - - - - -- - - - - - - SA SA SA SA SA SA SA SA - - - -- - -- - -- - - -- --------SA - - - - - - - - - - - - - - - - - - - - - - -- - -- - - -- - -- - -- - --------- - - - - - - - - - - - - - - - - - - SA - - - - - - - - - - - - - - - - - - - SA SA RS - SA SA SA SA - - - - - - - -- - - - - - - - - - - SA - SA SA SA - 4110 - - - - - - - - - - - - - - - - - - --- SA SA SA - 4111 - -- - - -- - -- - - -- -- -------- 4112 - - - - - --- - - - - - - -- - - - - - - - - - - TC Tonic-clonic/Cloniconvulsion,LY Lying, SA Salivation,RS : Reddishsalivation,BP :'Bradypnea, No abnormality,observed. Sex Dose level Animal (mg/kg) number 0107 0 0112 4107 4108 4109 Male 30 4110 4111 4112 Appendix 1-M-2 Clinicalsigns- Individual Test article Sample D-1 Sex Male ' Study Days after comencesent/cessationof treatment 28/0 29/1 30/2 31/3 32/4 33/5 34/6 35/7 36/8 37/9 38/10 39/11 4( RI DD Tt LY BP RD BU Rg Reduced spontaneous movement, TC Tonic-clonic/Clonicconvulsion.LY Lying, RD Reddish snivel,BP B BU Brownish urine, DD : Death. No abnormalityobserved. 2 Sex Dose level (mg/kg) 0 0.1 Female io Animal number 0201 4 0212 1201 4 1206 2201 4 2206 3201 4 3206 No abnormalityobserved. Appendix 1-F-1 Clinicalsigns- Individual Testarticle Sample D-1 Sex Female Study N Days after commencement 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 -- - -- - - -- - -- - -- - -- - - -- - - - -- - -- - - - -- -- -- - -- - - - -- - - - -- - -- - - -- - -- - -- - - -- - - - -- - -- - - -- - -- -- - - -- - -- - - - 3 Sex Dose level Animal (mg/kg) number 4201 4202 4203 4204 Female 30 4205 4206 4207 4208 4209 4210 Appendix 1-F-2 Clinicalsigns- Individual Test article Sample D-1 Sex Female Study Days after mencement 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 - - - - - - - - - - - - - - - - SA - - SA SA - SA - - - - - - - --- - - 7 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - SA - - SA SA - - - - - - - --- - - - - - SA SA - - SA SA SA SA SA - Rk TC CR CY --- - -- - - -- - - - -- --------- - - - - - - - - -- - - - - - - - - - - - - - SA SA --------- LU LU LU LO LilLH LU SA SA SA SA - - - - - - - - - - - - - - - - - Lu LB LU Lu LU Lu LU SA - - - - - - - - - ----- - - - - - LH LU LilLilLU LU LU - - - - - - - - - - - - - - - - - cs cs cs cs cs cs a 4211 -- - -- -- - - -- - - - -- -------- 4212 - - - - - - - - - - - - - - - - - - - - RI RX - - TC CR Rk Reduced spontaneous movement, TC To'nic-clonic/Clonciocnvulsion,LY : Lying, CR : Crouching,SA Salivati RS Reddishsalivation,CY : Cyanosis,LU : Loss of hair, CS : Coat staining, No abnormalityobserved. 4 Sex Dose level (mg/kg) 0 Female 30 Animal number 0207 0212 4207 4208 4209 4210 4211 4212 28/0 29/1 Appendix 1-F-3 Clinicalsigns- Individual Testarticle Sample D-1 Sex Female Study Days after commencement/cessationof treatment 30/2 31/3 32/4 33/5 34/6 35/7 36/8 37/9 38/1039/114 LH LU Lli LD LU Lli Lli LU Lil Lfl LH LH LU LU LH Lil LU Lil LU LU LH LH LU LH Lll LH Lll Lll LU LH Lli LU LH Lff LH LU cs cs cs cs cs cs cs LH Loss of hair,CS Coat staining. No abnormalityobserved. 5 Appendix 2-M-1 Body weight - Individual values Sex Male Dose level Sample D-1 0 mg/kg Study No. Animal Weeks after commencement/ cessation of treatment Number 0 1 2 3 4 6/1, 0101 0102 0103 0104 0105 0106 0107 0108 0109 0110 0111 0112 139 213 268 313 354 KS 142 196 249 291 318 KS 136 196 252 301 334 KS 140 207 265 3,18 352 KS 145 208 274 329 377 KS 148 215 275 327 370 KS 146 210 263 300 329 345 147 207 257 299 331 363 151 206 263 319 361 401 149 211 268 306 327 352 132 185 231 275 304 324 143 198 259 308 346 378 Mean 143 204 S.D. 5.6 8.8 N 12 12 KS : Killed on schedule 260 12.2 12 307 15.4 12 6 342 21.8 12 361 26.8 6 Appendix 2M 2 Body weight - Individual values Sex Male Dose level Sampl.e D-1 Study No 0.1 mg/kg Animal Weeks after commencement/ cessation of treatment Number 0 1 2 3 4 5/1 1101 137 206 277 333 375 KS 1102 140 200 244 288 321 KS 1103 141 210 268 318 362 KS 1104 146 199 254 293 321 KS 1105 148 214 284 340 376 KS 1106 148 214 285 347 399 K Mean 143 207 S.D. 4.6 6.6 N 6 6 ES : Killed on schedule 269 16.7 6 320 24.7 6 359 31.8 6 Appendix 2-M-3 Body weight - IndividuAl values Sex Male Dose level Sample D-1 1 mg/kg Study No. Animal Weeks after commencement/ cessation of treatment Number 0 1 2 3 4 5/1 2101 2102 2103 2104 2105 2106 136 197 255 308 346 KS 143 206 265 315 346 KS 142 202 264 296 354 KS 146 203 254 311 321 KS 147 213 278 336 376 KS 147 216 275 318 350 KS Mean 144 206 S.D. 4.2 7.1 N 6 6 KS : Killed on schedule 265 314 349 9.9 13.2 17.7 6 6 6 .Appendix 2 M -4 Body weight - Individual values Sex Male Dose level Sample D-1 10 mg/kg Study No. Weeks after commencement/ cessation of treatment Animal Number 0 1 2 3 4 5/1 3101 135 196 245 293 326 KS 3102 137 193 252 298 341 KS 3103. 143 195 245 289 320 KS 3104 145 1198 253 301 336 KS 3105 150 214 282 343 389 KS 3106 147 210 264 308 345 KS Mean S.D. N 143 5.8 6 KS Killed on schedule 201 8.8 6 257 14.2 6 305 19.6 6 343 24.5 6 Appendix 2M 5 Body weight - Individual values, Sex Male Dose level Sample D-1 30 mg/kg Study No. Animal Weeks after commencement/ cessation of treatment Number 0 1 2 3 4 5/1 4101 140 193 237 273 300 KS 4102 139 185 229 265 286 KS 4103 144 200 254 300 321 KS 4104 147 198 241 268 283 KS 4105 143 190 231 256 269 KS 4106 146 196 247 285 293 KS 4107 147 193 235 265 284 304 4108 136 189 233 268 291 311 4109 145 193 231 256 261 D 4110 138 180 228 263 284 308 .4111 142 194 238 271 292 315 4112 146 207 260 297 310 338 mean 143 193 S.D. 3.7 7.0 N 12 12 KS Killed on scheduleD Dead. 239 10.2 12 272 14.5 12 10 290 16.2 12 315 13.4 5 Appendix 2F1 Body weight - Individual values Sex Female Dose level. Sample D-1 0 mg/kg Study No. Weeks after commencement/ cessation of treatment Animal Number 0 1 2 3 4 5/1 0201 128 162 191 211 228 KS 0202 123 151 177 193 214 KS 0203. 131 160 181 207 221 KS 0204 140 178 208 235 264 KS 0205 128 164 186 207 227 KS 0206 0207 0208 0209 0210 131 165 138 178 131 168 129 168 128 171 187 214 227 KS 222 255 279 304 201 222 237 254 188 214 237 256 200 228 243 261 0211 133 171 196 220 244 262 0212 121 156 179 201 225 245 Mean -130 166 S.D. 5.4 8.1 N 12 12 KS : Killed on schedule 193 13.1 12 217 16.5 12 11 237 18.6 12 264 20.7 6 Appendix 2F2 Body weight - Individual values Sex Female Dose level Sample D-1 Study No. 0.1 mg/kg Animal Number 0 Weeks after commencement/ 1 2 cessation of treatment 3 4 5/1 1201 124 159 188 210 224 KS 1202 125 165 180 196 212 KS 1203 132 165 192 219 236 KS 1204 130 164 192 222 242 KS 1205 128 165 194 217 227 KS 1206 132 163 189 207 222 KS Mean 129 164 189 212 227 S.D. 3.4 2.3 S.o 9.6 10.5 N 6 6 6 6 6 KS : Killed on schedule 12 Appendix 2F3 Body weight - Individual values Sex Female Dose level Sample D-1 I mg/kg Study No. B u Animal Number 2201 2202 2203. 2204 2205 2206 Mean S.D. N KS Kiiieu 0 121 128 130 132 134 141 -131 6.6 6 on scnectuie Weeks after commencement/ 1 2 cessation of treatment 3 4 5/1 152 177 204 225 KS 167 184 205 226 KS 157 180 201 216 KS 171 202 228 240 KS 185 213 237 259 KS 180 198 225 246 KS 167 13.7 6 192 14.2 6 217 15.2 6 235 15.9 6 13 Appendix 2F4 Body weight - Individual values Sex Female Dose level Sample D-1 10 mg/kg Study No. Weeks after commenc6ment/ cessation of treatment Animal Number 0 1 2 3 4 5/1 3201 125 156 185 203 216 KS 3202 125 164 193 218 225 KS 3203 125 149 166 178 186 KS 3204 130 159 189 208 221 KS 3205 136 169 187 203 212 KS 3206 138 169 191 208 223 KS Mean 130 161 S.D. 5.9 7.9 N 6 6 KS Killed on schedule 185 203 214 9.8 13.4 14.4 6 6 6 14 Appendix 2F5 Body weight - Individual values Sex Female Dose level Sample D-1 30 mg/kg Study No. Animal Number 0 Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 4201 131 160 174 179 178 KS 4202 130 156 169 180 194 KS 4203 127 151 172 186 194 KS 4204 136 168 184 179 204 KS 4205 133 16*2 173 177 181 KS, 4206 135 159 172 182 191 KS- 4207 129 157 167 162 175 194 4208 128 144 158 161 169 188 4209 131 160 177 181 190 196 4210 129 159 171 169 168 183 4211 122 149 159 175 186 197 4212 134 167 175 179 187 212 Mean 130 158 171 176 185 195 S.D. 3.9 7.0 7.2 7.8 10.9 9.9 N 12 12 12 12 12 6 KS : Killed on schedule 15 Appendix- 3 m- 1 Food consumption Sex : Dose level : Individual values Mate Sample D-1 0 mg/kg Study No. Unit gl Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 6/1 6/2 1 17.8 20.4 18.6 16.3 NA 2 17.0 19.2 19.6 18.8 NA 3 16.9 19.6 19.6 17.4 NA 4 17.4 19.1 17.6 15..0 20.6 22.3 5 16.1 18.5 19.2 17.1 23.1 23.8 6 15.5 17.7 18.5 16.4 21.6 23.0 Mean S.D. N 16.8 0.85 6 NA No animal existed. 19.1 0.92 6 18.9 0.77 6 16.8 1.27 6 21.8 1.26 3 23.0 0.76 3 16 .Appendix 3M .Food consumption Sex : Dose level : 2 Individual values Male Sample D-1 .0.1 mg/kg Study No. Unit : g/A Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 17.8 20.2 19.5 18.1 NA 2 16.8 18.9 17.6 16.7 NA 3.. 19.3 22.6 23.4 21.6 NA Mean S.D. N 18.0 1.26 3 NA No animal existed. 20.6 1.88 3 20.2 2.96 18.8 2.52 3 17 Appendix 3M Food consumption Sex : Dose level : 3 Individual values Male Sample D-1 I mg/kg Study No. Unit gl Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 16.3. 18.8 18.1 17.3 NA 2 16.9 19.1 17.1 16.6 NA 3 17.8 19.9 19.1 17.9 NA Mean S.D. N 17.0 0.75 3 NA No animal existed. 19.3 0.57 3 18.1 1.00 3 17.3 0.65 3 18 Cage Number 1 1 15.7 2 16.0 3 18.0 Mean s.b. N 16.6 1'.25 3 NA No animal existed. Appendix 3M 4 Food consumption Sex : Dose level : Individual values Male Sample D-1 10 mg/kg Study No. Unit gll Weeks'after commencement/ cessation of treatment 2 3 4 5/1 6/2 17.1 16.9 15.9 NA 17.6 19.1 16.7 NA 20.1 22.4 20.8 NA 18.3 1.61 3 19.5 2.77 3 17.8 2.63 3 19 Appendix 3-M Food consumption Sex : Dose level : 5 Individual values Male Sample D-1 30 mg/kg Study No. Unit gl Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 14.8 14.9 14.7 12.8 NA 2 15.8 16.3 15.6 12.8 NA 3 15.8 16.4 16.3 11.7 NA 4 15.1 15.2 15.6 13'.7 17.8 20.2 5 14.5 15.4 15.0 12.5 22.7 19.7 6 16.4 16.4 15.4 12.8 20.6 23.6 Mean S.D. N 15.4 0.72 6 NA No animal existed. 15.8 0.68 6 15.3 0.36 6 12.7 0.64 6 20.4 2.46 3 21.2 2.12 3 20 Appendix 3-- F Food consumption Sex : Dose level : 1 Individual values Female Sample D-1 0 mg/kg Study No. Unit g/A Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 12.4 2 14.7 3. 12.6 4 14.8 5 14.9 6 13.4 12.1 13.6 11.4 14.9 14.3 12.4 12.3 15.1 12.4 15.6 15.1 13.1 11.2 13.8 10.8 14.-3 13.3 12.9 NA NA NA 19,6 19.2 19.4 20.4 20.0 19.7 Mean S.D. N 13.8 1.15 6 NA : No animal existed. 13.1 1.36 6 13.9 1.50 6 12.7 1.42 6 19.4 0.20 3 20.0 0.35 3 21 Appendix 3F2 Food consumption Sex : Dose level : Individual values Female Sample D-1 0.1 mg/kg Study No. Unit g/ Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 13.3 11.8 12.1 11.4 NA 2 14.1 13.4 14.9 13.6 NA 3 13.6 12.7 13.1 12.1 NA. Mean S.D. N 13.7 0.40 3 NA No animal existed. 12.6 0.80 3 13.4 1.42 3 12.4 1.12 3 22 Appendix 3F3 Food consumption Sex : Dose level : Individual values Female S ample D-1 1 mg/kg Study No. Unit gl Cage Number Weeks after commencement/ cessation o.f treatment 1 2 3 .4 5/1 6/2 1 11.6 10.5 11.7 11.0 NA 2 14.1 12.9 13.1 12.2 NA 3 16.3 13.4 13.8 12.5 NA Mean S.D. N 14.0 2.35 3 -12.3 1.55 3 NA No animal existed. 12.9 1.07 3 11.9 0.79 3 23 Appendix 3F Food consumption Sex : Dose level : 4 Individual values Female Sample D-1 10 mg/kg Study No. Unit g/ Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 12.6 11.9 11.2 9.6 NA 2 11.6 .10.6 10.4 9.4 NA 3 13.8 12.1 11.9 10.1 NA Mean S.D. N 12.7 1.10 3 11.5 0.81 3 NA No animal existed.. 11.2 0.75 3 9.7 0.36 3 24 Appendix 3 F 5 Food consumption Sex : Dose level : Individual values Female Sample D-1 30 mg/kg Study No. Unit g/A Cage Number Weeks after commencement/ cessation of treatment 1 2 3 4 5/1 6/2 1 12.0 9.6 7.9 6.9 NA 2 13.4 11.3 8.5 9.6 NA 3 13.4 10.6 9.0 8.6 NA 4 11.9 9.8 6.6 8.5 14.1 14.6 5 13.7 10.7 7.9 6.8 12.9 13.9 6 13.2 9.5 8.4 7.6 15.9 17.6 Mean S.D. N 12.9 0.78 6 NA : No animal existed. 1,0.3 0.72 6 8.1 0.82 6 8.0 1.09 6 14.3 1.51 3 16.4 1.97 3 25 Al)l)eixid 4 - tl- I Stud Hematology Individualvalues Sex tlale Dose level Sample D-1 0 mg/kg Animalskilledon seliedul(e4 weeks) Animal Number 0101 0102 0103 0104 0105 0106 tlean S.D. N RBC HB Ht couti conc. (XIOI/MM3) (g/di) % 774 14.6 50.5 780 13.8 50.6 681 13.7 44.4 775 14.6 45.6 695 14.2 47.3 705 14.0 45.6 735 14.2 47.3 46.0 0.39 2.66 6 6 6 mcv (gNe) 65 65 65 59 68 65 65 2.9 6 MCH (pg) 18.9 17.7 20.1 18.8 20.4 19.9 19.3 1.02 6 tichc Platelet PT AP count % (Xlol/M3) (sec) (s 28.9 84.8 13.8 14 27.3 104.6 14.1 16 30.9 102.6 14.1 17 32.0 115.4 13.8 16 30.0 88.2 14.9 15 30.7 83.7 15.2 18 30.0 96.6 14.3 16 1.66 12.88 0.59 1. 6 6 6 26 Append ix 4 M - 2 Stu llematology Individualvalues Sex Male Dose level Sample 0-1 0.1 mg/kg Animalskilledon selledul(e4 weeks) Animal Number 1101 1102 1103 1104 1105 1106 Mean S.D. N RBC HB Ht count conc. (XI04/MM3) (g/di) % 765 14.1 52.2 683 13.9 46.0 689 13.9 45.4 675 13.8 44.8 679 14.3 46.1 696 14.1 45.6 698 14.0 46.7 33.7 0.18 2.74 6 6 6 mcv Am') 68 67 66 66 68 66 67 1.0 6 MCH (pg) 18.4 20.4 20.2 20.4 21.1 20.3 20.1 0.91 6 MCHC Platelet PT Af count % (xlo'@/mm, (see) 27.0 106.1 14.9 1 30.2 78,;2 13.9 1 30.6 8B.'3 13.9 1 30.8 91.0 14.0 1 31.0 94.7 13.8 1 30.9 104.7 13.7 1 30.1 93.8 14.0 1 1.54 10.51 0.44 1 6 6 6 27 Appendix 4 M 3 s Hematology- Individualvalues Sex tlale Dose level Sample U-1 I mg/kg Animalskilledon scliedul(e4 weeks) An iinaI Number 2101 2102 2103 2104 2105 2106 Mean S.D. N RBC HB Ht count conc. (XIO@/nIM3) (g/dl) % 681 13.2 42.3 733 14.0 46.3 794 14.1 51.9 764. 14.3 47.7 719 13.8 46.2 728 14.4 49.0 737 14.0 47.2 38.8 0.43 3.21 6 6 6 mcv m, 62 63 65 62 64 67 64 1.9 6 mcii (pg) 19.4 19.1 17.8 18.7 19.2 19.8 19.0 0.69 6 tichc % Platelet count (XI04/MM3) PT (sec) 31.2 97.4 13.9 30.2 92.1 14.2 27.2 134.7 14.1 30.0 112.7 14.1 29.9 105.4 14.4 29.4 79.8 13.5 29.7 103.7 14.0 1.34 18.02 0.31 6 6 Aljl)eii.i\d 4 tl.-4 Stud Hematology- Individualvalues Sex Male Dose level Sample D-1 10 mg/kg Animalskilledon schedule(4 weeks) RBC HB lit Animal count Colic. Nunibei, (XI04/mm') (&/di) % 3101 681 12.8 43.4 3102 714 13.6 45.1 3103 737 13.5 49.6 3104 756 13.3 45.4 3105 713 13.7 46.2 3106 766 13.2 47.6 Mean 728 13.4 46.2 S.D. 31.4 0.33 2.15 N 6 6 6 mcv /IrA3) 64 63 67 60 65 62 64 2.4 6 MCH (pg) 18.8 19.0 18.3 17-6. 19.2 17.2 18.4 0.80 6 ticuc % Platelet count (x104/M3) PT (sec) AP (S( 29.5 97.5 14.4 14 30.2 111.8 13.9 14 27.2 84.3 13.6 14 29.3 115.5 13.8 17 29.7 100.9 14.0 17 27.7 lll-9 13.6 15 28.9 103.7 13.9 15 1.20 11-79 0.30 1. 6 6 6 29 Appendix 4 M - 5 su Hematology Individualvalues Sex. Male Dose level SamlileD-1 30 mg/kg Animalskilledon schedule(4 weeks) Animal Number 4101 4102 4103 4104 4105 4106 Mean S.D. N RBC HR [it count COIIC. (XI04r/MM3) (g/di) % 647 13.1 44.1 665 12.5 43.8 702 13.0 45.9 725 12.9 48.4 667 13.6 43.0 792 14.9 51.6 700 13.3 46.1 53.3 0.85 3.30 6 6 6 mcv jIM3) 68 66 65 67 64 65 66 1.5 6 MCH (pg) 20.2 18.8 18.5 17.8 20.4 18.8 19.1 1.01 6 fichc Platelet PT A count % (XI04/MM3) (sec) 29.7 92.1 14.4 1 28.5 .88.1 14.8 2 28.3 96.0 14.6 1 26.7 96.3 14.2 1 31.6 78.0 14.0 2 28.9 89.1 14.2 1 29.0 89.9 14.4 1 1.63 6.76 0.29 2 6 6 6 30 Apl)eiixd 4 M 6 Stu liematology Individualvalues Sex Male Dose level Sample D-1 0 mg/kg Animalskilledon schedule(Recovery) ROC flu lit Animal count conc. Numbei- (XIO@/MM3)(g/di) % 0107 790 14.5 45.0 0108 728 13.4 40.4 0109 798 14.2 46.6 0110 896 14.3 48.2 0111 784 13.8 45.2 0112 791 14.1 47.7 Mean 798 14.1 45.5 S.D. 54.4 0.39 2.82 N 6 6 6 CL : Not measured becauseclottiiig. mcv (Am,) 57 55 58 54 58 60 57 2.2 6 m@ii (pg) 18.4 18.4 17.8 16.0 17.6 17.8 17.7 0.88 6 ticiic Platelet PT AP count % (XIOI/=3) (sec) (s 32.2 71.4 33.2 97.0 30.5 71.0 29.7 97.6 30.5 80.2 29.6 113.1 13.8 17 14.0 16 13.8 15 13.8 15 CL 13.7 15 31.0 88.4 13.8 16 1.44 16.90 0.11 0. 6 6 5 31 Appendix 4 M 7 Stu tlematolog-y Individualvalues Sex Male Bose level Sample D-1 30 mg/kg Animalskilledon seliedul(eRecovery) Animal Number RBC llu Ht count Colic. (xIO4/mm') (g/dl) % 4107 743 12.7 43.8 4108 688 11.9 39.6 4109 D 4110 718 12.5 41.5 4111 703 13.2 42.7 4112 735 13.0 40.5 Mean 717 12.7 41.6 S.D. 22.6 0.50 1.68 N 5 5 5 D : Dead. @lcv /1M3) 59 58 58 61 55 58 .2.2 5 tit'n (pg) 17.1 17.3 17.4 18.8 17.7 17.7 0.67 5 ticuc Platelet PT A count % (Xlolt/=3) (sec) 29.0 82.8 13.3 1 30.1 112.7 13.7 1 30.1 96.0 13.7 1 30.9 88.6 12.6 1 32.1 97.4 13.6 1 30.4 95.5 13.4 1 1.15 11.28 0.47 1 5 5 5 32 Appendix 4 F I Stud Hematolo&y Individualvalues Sex Female [)oselevel Saini)lDe-1 0 mg/kg Animals killed on scliedule(4 weeks) Animal Number 0201 0202 0203 0204 0205 0206 Mean S.D. N RBC HB Ht count colic. (XI04/MM3) (g/di) % 720 13.2 44.5 777 13.7 46.3 658 13.3 40.6 635 12.6 37.6 727 13.4 41.8 714 13.8 45.7 705 51.2 6 13.3 0.43 6 42.8 3.36 6 mcv itM3) 62 60 62 59 57 64 61 2.5 6 mcii (pg) 18.3 17.6 20.2 19.8 18.4 19.3 18-l') 1.00 6 tichc Platelet PT AP count % (XI04/=3) (se'c) (se 29.7 29.6 32.8 33.5 32.1 30.2 117.7 142.0 95.2 107.5 120.4 111.4 14.3 14.4 14.6 14.2 14.0 14.3 15. 13. li. 12. 15. 14. 31.3 115.7 14.3 14. 1.70 15.64 0.20 1.( 6 6 6 33 Appendix 4 F 2 Stu Hematology- Individualvalues Sex Female Dose level Sample D-1 0.1 mg/kg Animalskilledon schedule(4 weeks) Animal Number 1201 1202 1203 1204 1205 1206 Mean S.D. N RBC HB Ht count conc. (XI04/MM3) (g/dl) % 720 13.5 40.7 693 13.5 40.8 644 13.4 42.6 699 13.9 41.3 698 13.7 42.3 693 13.5 44.4 691 13.6 42.0 25.2 0.18 1.40 6 6 6 mcv (AM3) 57 59 66 59 61 64 61 3.4 6 MCH (pg) 18.8 19.5 20.8 19.9 19.6 19.5 19.7 0.66 6 MCHC Platelet PT AP count % (XI04/=3) (sec) (s 33.2 101.4 14.7 11 33.1 97.6 14.5 14 31.5 124.0 14.5 If 33.7 87.7 13.4 l@ 32.4 99.1 14.2 1 30.4 108.6 14.2 1 32.4 103.1 14.3 1 1.24 12.27 0.46 6 6 6 34 Alipeiiidx 4 F 3 Stud Hematology Iiidividiivaallues Sex Female Dose level Sample D-1 I mg/kg Animalskilledon seliedule(4 weeks) Animal Number 2201 2202 2203 2204 2205 2206 Mean S.D. N RBC HB Ht count CORC. (XI04/MM3) (g/di) % 726 13.7 42.6 708 12.9 40.6 724 13.5 44.0 634 13.2 40.7 768 13.1 41.3 643 13.3 38.8 701 13.3 41.3 52.0 0.29 1.79 6 6 6 M('V M3) 59 57 61 64 61 60 60 2.3 6 MCH (pg) 18.9 18.2 18.6 20.8 19.3 20.7 19.4 1.09 6 MCHC Platelet PT AP count % (XI04/MM3) (sec) (s 32.2 106.3 13.9 13 31.8 115.6 14.2 16 30.7 113.6 14.8 15 32.4 lti-I 13.2 14 31.7 102.7 14.4 17 34.3 97.9 14.6 17 32.2 107.9 14.2 15 1.19 6.81 0.57 1. 6 6 6 35 Appentlix 4 - F - 4 Stud Ilematolog-y Individualvalues Sex Female Dose level SamiileD-1 10 mg/kg Animalskilledon scliedul(e4 weeks) RBC HB Ht Animal count conc. Number (XI04,/MM3) (g/di) % ticv AM3) tich (118) MCHC tllatelet PT AP count % (XI04/BMI) (sec) (se 3201 685 14.0 44.0 64 20.4 31.8 88.0 14.3 15. 3202 759 13.5 43.4 57 17.8 31.1 111.0 14.2 14. 3203 764 14.1 47.4 62 18.5 29.7 123.3 14.1 15. 3204 700 13.4 41.0 59 19.1 32.7 93.7 13.5 14. 3205 693 13.5 43.4 63 19.5 31.1 117.7 13.5 15. 3206 695 13.9 i43.8 63 20.0 31.7 101.9 14.5 16. Mean 716 13.7 43.8 S.D. 35.6 0.30 2.06 N 6 6 6 61 19.2 31.4 105.9 14.0 15. 2.7 0'.96 1.00 13.81 0.42 0.9 6 6 6 6 6 36 Appeiv]ix 4 F 5 Hematology Individualvalues Sex Female Dose level Sample D-1 30 mg/kg Animalskilledon schedule(4 weeks) Study Animal Number 4201 4202 4203 4204 4205 4206 tleati S.D. N RBC HB Ht. count conc. (xIO4/mm3) (g/di) % 778 13.9 44.7 649 12.9 39.6 763 12.8 46.6 713 12.7 -'42.5 666 12.5 39.9 761 12.9 45.5 722 13.0 43.1 54.6 0.49 2.95 6 6 6 mcv. (,um, 57 61 61 60 60 60 60 1.5 6 tich (pg) 17.9 19.9 16.8 17.8 18.8 17.0 18.0 1.16 6 MCHC Platelet PT count yo (XI04/11,@3) (sec) 31.1 95.6 13.7 32.6 97.3 13.5 27.5 117.9 13.3 29.9 1. 01.7 13.3 31.3 90.4 13.5 28.4 116.4 13.1 30.1 103.2 13.4 1.92 11-39 0.21 6 6 6 APTT (sec) 16.6 15.2 11.4 16.9 14.6 13.0 14.6 2.12 6 37 Appendix 4 F - 6 Hematology Individualvalues Sex Female Dose level Sample D-1 0 ms/kg Animals killedon schedule(Recovery) Study Animal Number 0207 0208 0209 0210 0211 0212 Mean S.D. N RBC HR "t count CORC. (xIO4/wO) (g/dl) % 713 13.9 42.1 727 13.6 42.0 683 13.4 41.1 755 14.1 44.4 699 13.4 .43.2 713 13.7 45.6 715 13.7 43.1 24.6 0.28 1.68 6 6 6 ticv (ILMI) 59 58 60 59 62 64 60 2.3 6 @ICH (pg) 19.5 18.7 19.6 tB.7 19.2 19.2 19.2 0.38 6 mcilc % Platelet count (XI04/MM3) PT (sec) 33.0 112.4 13.3 32.4 90.8 14.0 32.6 88.3 13.3 31.8 99.8 13.2 31.0 72.5 13.9 30.0 99.1 13.2 31.8 93.8 13.5 1.12 13-44 0.37 6 6 6 APTI (se( 15.1 17.E 13.( 14.5 16.1 15.9 15.5 1.62 6 38 Appendix 4 - P - 7 Stud Hematology Individualvalues Sex Female Dose level Sample D-1 30 mg/kg Animalskilledon schedule(Recovery) Animal Number 4207 4208 4209 4210 4211 4212 tlean S.D. N RBC IIB Ht count conc. (XI04/MM3) (g/di) % 666 12.5 41.4 700 12.6 44.6 641 12.2 41.2 653 12.6 41.2 732 13.0 44.4 678 12.9 43.3 678 12.6 42.7 33.3 1.62 6 6 6 mcv I.LM3) 62 64 64 63 61 64 63 1.3 6 MCH (pg) 18.8 18.0 19.0 19.3 17.8 19.0 18.7 0.61 6 tichc Platelet PT API' count % (XI04/MM3) (sec) (se 30.2 114.4 13.4 13. 28.3 138.9 13.8 12. 29.6 85.4 13.2 12. 30.6 .105.3 12.8 13. 29.3 101.4 13.0 10. 29.8 116.5 12.5 29.6 110.3 13.1 12. 0.80 17.87 0.46 1.3 6 6 6 39 Animal Number 0101 0102 0103 0104 0105 0106 Mean S.D. N Appendix 5m I Hematology Individual values Sex Male Dose.level Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) Study No. WBC count (xlO?/mm3) Differential count of leukocytes of total counted c Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 69 83 9 1 1 0 83 91 7 1 0 0 119 90 4 1 0 0 63 90 6 0 0 0 80 77 13 0 1 0 ill 82 10 0 0 0 88 86 8 1 0 0 22.6 5.7 3.2 0.5 0.5 0.0 6 6 6 6 6 6 .40 A.nimal Number 1101. 1102 1103 1104 1105 1106 Mean S.D. N Appendix 5 M-2 Hematology - Individual values Sex Niate Dose level Sample D-1 O.i Animals kil,led on sctiedule (4 weeks) mg/kg Study No. WBC count (Xlop-/MM3) Differential count of leukocytes of total counted c Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 120 82 15 0 0 0 86 88 6 1 0 0 94 80 9 1 3 .0 126 93 6 0 0 0 135 93 4 0 0 0 73 83 10 0 1 0 106 87 8 0 1 0 24.8 5.7 3.9 0.5 1.2 0.0 6 6 6 6 6 6 41 Animal Number 2101 2102 2103 2104 2105 2106 Mean S.D. N Appendix 5m 3 Hematology Individual values Sex Male Dose level Sample D-1 1 mg/kg Animals killed on schedule (4 weeks-) Study No. WBC count (xlO2/mm") Differential count of leukocytes of total counted c Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phi'ls phils 104 95 3 0 1 0 109 89 7 0 0 162 89 8 0 0 0 iii 87 9 0 1 6 116 82 12 1 0 0 96 86 10 0 1 0 118 88 8 0 1 0 231,3 @4.3 3.1 0.4 0.5 0.0 6 6 6 6 6 112 Animal Number 3101 3102 3103 3104 3105 3106 Mean -S.D. N 'Appendix 5M 4 Hematology Individual values I Sex Male Dose level Sample D-1 10 Animals killed on schedule (4 weeks') mg/kg Study No. WBC count (Xlo?-/MM3) Differential count of leukocytes of total counted LymphoCyteS Neutrophils Segmented Band Eosinophils Basophils 105 78 16 0 1 0 107 82 10 .0 0 0 123 82 9@ 0 2 0 86 92 6 0 0 0 81 82 12 0 0 0 80 91 6 0 2 0 97 85 10 0 1 0 17.4 5.6 3.5 0.0 1.0 0.0 6 6 6 6 6 6 43 Animal Number '4101 4102 4103 4104 4105 4106 Mean S.D. N Appendix 5m 5 Hematolog.y Individual values Sex Mate Dose level Sample D-1 30 Animals killed on schedule (4 weeks) mg/kg Study No. WBC count (XJ02/MM3) 245 108 104 112 66 184 137 05.5 6 Differential count of leukocytes of total counted Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils. phils 91 6 0 0 0 86 7 0 0 0 88 9 2. 0 0 84 8 1 0 0 85 13. 0 0 0 84 9 0 0 0 86 9 1 0 0 2.8 2.4 0.8 0.0 0.0 6 6 .6 6 6 44 Animal Number 0107 0108 0109 0110 0111 0112, Mean S.D. N Appendix 5M 6 Hematology Individual values Sex Male Dose level Sample D-1 0 mg/kg Animals killed on schedule (Recovery) Study No. WBC count (xlO@/mm3) Differential count of leukocytes. of total counted Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 96 90 4 1 0 0 96 75 20 .0 0 0 117 90 5 1 0 0 78 79 14 0 0 106 90 8 0 0 0 86 83 12 0 0 0 97 85 11 0 0 0 13.9 6.5 6.1 0.5 0.0 0.0 6 6 6 6 6 6 45 Appendix 5M7 Hematology - Individual values Sex Male Dose level Sample-D-1 30 Animals killed on schedule (Recovery) mg/kg Study No., Animal Numbei-. WBC count (XIOP/MM3) 4107 4106 4109 4110 4111 4112 Mean S.D. N D Dead.. 122 134 D 107 130 135 126 11.6 5 Differential count of leukocytes % of total counted Lympho- Neutraphils Eosino- Baso- cytes Segmented Band phils phils 87 8 1 0 0 86 8 1 0 0 83 7 1 0 0 85 10 0 0 0 95 5 0 0 0 87 8 1 0 0 4.6 1.8 0.5 0.0 0.0 5 5 5 5 5 46 Animal Number 0201 0202 0203 0204 0205 0206 Mean S.D. N Appendix 5F 1 Hematology Individual values Sex Female Dose level Sample D-1 0 Animals killed on schedule (4 weeks) mg/kg Study No. WBC count (Xlo2/MM3) 54 80 77 80 72 84 75 10.8 6 Differential count of leukocytes of total counted c Lympho-. Neutrophils Eosino- Baso- cytes Segmented Band phils phils 86 10 1 0 92 8 -.0 0 0 85 13 0 0 0 87 9 1 0 0 84 12 0 0 0 84 8 2 0 0 86 10 1 0 0 3.0 2.1 0.8 0.0 0.0 6 6 6 6 6 47 Animal Number 1201 1202 1203 1204 1205 1206 Mean S.D. N Appendix 5F2 Heinatol(igy Individual values Sex Female Dose level Sample D-1 0.1 Animals killed on schedule (4 weeks) mg/kg Study No. WBC count (Xlo2./MM3) 143 58 129 .96 86 92 ioi 30.7 Differential count of leukocytes % of total counted c Lympho- Neutrophils" Eosino- Baso- cytes Segmented Band phils phils 86 9 .00 0 0 94 3 1 0 82 12 1 0 0 93 5 1 0 0 89 3 0 0 0 85 11 0 0 0 88 7 0 0 0 4.7 4.0 0.5 0.4 0.0 6 6 6 6 6 48 Animal Numbei- 2201. 2202 2203 2204 2205 2206 Mean S.D. N Appendix 5F3 Flematology Individual values Sex. Female Dose level Sample D-1 1 mg/kg Animals killed on schedule (4 weeks) Study No. WBC count (Xlo2/MM3) 57 94 108 112 77 70 86 21.9 6 Differential count of leukocytes of total counted Lymptio- Neutrophils Eosino- Baso- cytes Segmented Band ptiils phils 86. 9 1 0 0 92 7 0 0 87 10 0 1 0 81 15 0 1 0 86 13 0 0 0 93 5 1 0 0 88 10 0 0 0 4.@l 3.7 0.5 0.5 0.0 6 6 6 6 6 49 Animal Number 3201 3202 3203 3204 3205 3206 Mean S.D. N Appendix 5-F-4 liematology - Individual values Sex Female Dose level Sample D-1 10 .Animals killed on scliedule (4 weeks) mg/kg Study No. WBC count (Xlo2/MM3) 117 103 61 67 89 70 85 22.3 6 Differential count of leukocytes % of total counted Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 92 4 1 0 0 87 6 0 0 0 90 6 0 0 0 89 6 0 1 0 92 2 0 0 0 84 8 0 1 0 89 5 0 0 0 3.1 2.1 0.4 0.5 0.0 6 6 .6 6 6 50 Animal Number 4201 4202 4203 4204 4205 4206 Mean S.D. N Appendix 5 F 5 Hematology Individual values Sex Female Dose level Sample D-1 30 Animals killed on schedule (4 weeks) mg/kg Study No. WBC count (Xlo?/MM3) Differential count of leukocytes Lympho- Neutrophils cytes Segmented Band Y. of Eosinophils total counted Basophils 78 89 6. 0 0 0 66 83 7 .1 0 0 83 89 7 0 0 0 104 87 7 0 1 0 60 92 5 0 0 0 71 80 18 0 0 0 77 87 8 0 0 0 15.6 4.4 4.8 0.4 0.4 0.0 6 6 6 6 6 6 51 Animal Number @0207. 0208 0209 0210 0211 0212 Mean S.D. N Appendix 5 F-6 Hematology - Individual value7s Sex Female Dose level Sample D-1 0 mg/kg Animals killed on scliedule (Recovery) Study No. WBC count (XlU2/MM3) 69 92 54 114 72 109 85 23.9 6 Differential count of leukocytes % of total counted Lympho- Neutrophils Eosino- Baso- cytes Segmented Band phils phils 82 4 1 2 0 04 2 0 0 0 87 9 0 0 0 82 13 0 0 0 91 7 0 0 0 77 16 0 3 0 86 9 1 0 6.3 5.3 0.4 .1.3 0.0 6 6 6 6 6 52 Animal Number 4207 4208 4209 4210 4211. 4212 Mean S.D. N Appendix 5F7 Hematology Individual values Sex Female . Dose level Sample D-1 30 Animals killed on schedule (Recovery) mg/kg Study No. WBC count (xlO?/mm3) Differential count of leukocytes % of total counted Lympho- Neuti*ophils Eosino- Baso- cytes Segmented Band phils phils 105 9'2 3 1 0 0 104 88 6 0 0 0 84 88 5 0 1 0 76 93 4 0 0 0 88 91 8 0 0 0 57 93 2 0 0 0 86 91 5 0 0 0 18.1 2.3 2.2 0.4 0.4 0.0 6 6 6 6 6 6 53 Ai)i)eiiixd 6 - @l I s C Iiiiica I clieniisti,,v Iiltiiv idiiaI va Ities Sex tiaIe I)oselevel Samiiie U-1 0 mg/kg Animals killed on scliedule(4 weeks) Animal Ntiinber 0101 0102 0103 0104 0105 0106 tleaii S.D. N G(IT ILI-/I) GPT (IU/I) 80 37 70 26 60 33 64 26 65 - 31 71 26 68 30 7.0 4.6 6 6 r -GTI' (IU/I) 0 0 0 0 0 0 0 0.0 6 ALP ([U/I) 739 667 859 642 841 518 711 129.3 6 Lli-ea Ci-eatiiiiiiGlucose T iiit rogi@ii c (mg/di) (mg/dl) (mg/di) 11.2 0.5 134 13.4 o.5 129 14.8 0.5 153 12.2 0.5 138 12.6 0.5 i5o 10.2 0.5 137 12.4 0.5 140 1.62 0.00 9.4 6 6 6 54 Api)eiiidx 6 M 2 iiica I cliemisti,,\, I11(iiv i(itiIa va Ities S e.\ Male I)oselevel Saml)le[)-1 0 mg/kg Animals killed on schedule (4 weeks) Study No. B .Anima I Nunibei, Total protein (8/(Il) Albumin (g/di) 0101 6.36 4.07 0102 6.34 3.92 0103 6.63 4.00 0104 6.68 4.04 0105 6.27 3.75 0106 6.78 4.08 @leaii S.D. N 6.51 0.212 6 3.98 0.125 A/G ratio 1.78 1.62 1.52 1.53 1.49 1.51 1.58 0.110 6 Calcium (mg/di) Inorganic I)IIOS. (mg/dl) Na (mEq/1) 9.5 9.3 10.6 9.6 9.9 9.9 9.4 142 9.8 143 10.8 143 10.0 142 ll-.2 142 9.7 142 9.8 10.2 142 0.46 0.70 0.5 6 6 6 K (mEq/1) 4.0 4.1 4.1 4.2 4.0 4.1 4.1 0.08 6 55 Al)l)eii.d\ 6 ti 3 si C Iiiiica I cliemIstry Iiidiv i(itiIa va Ities Sex @laIe Dose level Sample 0-1 0.1 mg/kg Aiiimalskille(ioilscliedule(4 weeks) Allima I Ni.ifnbe 1, 1101 1102 1103 1104 1105 1106 tleaii S.D. N (;(IT (ILI1/) (15 77 69 61 66 50 65 U-0 (3i'T (1Li/I) 29 31 38 26 30 23 30 5.1 6 r -(;TP (IU/I) 0 0 0 0 0 0 0 0.0 6 ALP (IU/I) 822 666 713 577 618 666 677 84.9 6 L)i,ea Ci-eatiiiiiiGlucose T iiIt rogeii c (mg/di) (mg/dl) (mg/di) 11.8 0.4 136 13.2 0.5 138 10.1 0.4 157 11.5 0.4 149 10.4 0.5 159 6.8 0.5 159 10.6 2.18 6 0.5 0.05 6 150 10.5 6 56 Animal Number 1101 1102 1103 1104 1105 1106 tlean S.D. N Aijl)eiiixd 6 M 4 Study No. BM (,Iiiiicalcliemistry Ilidivilitiavlalues Sex Male Dose level Sample U-1 0.1 mg/lig Aliima Is k iIIe(ion sclietiIeti(4 weeks) To t,,It protein (g/di) 6.29 6.67 6.21 6.31 6.48 6.33 6.38 0.166 6 A Ilitiimii (-O/dl) 3.95 4.16 3.93 3.90 3.90 3.89 3.96 0.103 6 A/(; ratio 1.69 1.66 1.72 1.62 1.51 1.59 1.63 0.076 6 Calcium Inorganic Na K plios. (mg/dl) (mg/dl) (mEq/1) (mEq/1) (m 9.6 9.3 142 3.9. u-9 9.0 142 4.1 10.1 10.4 142 4.1 9.8 8.8 142 4.0 9.9 10.9 142 4.0 9.3 8.4 141 4.0 9.8 0.28 6 9.5 0.98 6 142 4.0 0.4 0.08 6 6 57 Aljl)ei.i\(I 6 @l 5 s C Iiiiica I cliemisti,y Iiitilv idiiaI va Iii.es Sex tiaIe I)oselevel Sample [)-I I mg/kg AiiiniaIs k iIIeilt)tsi(-IiedIeu (4 weeks) Aiiima I Ntinibei- 2101 2102 2103 2104 2105 2106 tleaii S.D. N (;flT (IV/I) (16 87 69 67 66 70 71 8.1 6 t.1;1T (IU/I) 26 36 34 33 30 25 31 4.5 6 t.-(iTP (iti/1) ALI' (IU/I) 0 705 0 923 0 537 0 806 0 727 0 703 0 734 0.0 127.7 6 6 Ui-ea Ci,eatiiiiii Glucose T iitrogeti c (mg/dl) (mg/di) (mg/dl) 11.3 111.1 12.3 12.6 16.8 15.5 0.5 145 0.5 129 0.5 143 0.5 136 0.5 139 0.4 -148 13.3 0.5 140 2.34 0.04 6.9 .6 0 6 C iiica I clieniisti-y Iiitiiv ititiIa va Iues Sex tiale I)oselevel Sami)le D-1 I lng/kg Animals killed on scliedule(4 weeks) Study No. Btl Ailima I Number 2101 2102 2103 2104 2105 2106 @leaii S.D. N Total A Ibtimiii A/G protein i-atio (g/(Il) (8/(Il) 6.10 6.36 6.35 6.07 6.43 5.95 3.88 3.92 3.82 3.93 3.77 3.74 1.70 .1.61 1.51 1.84 1.42 1.69 6.22 0.190 6 3.84 0.079 6 1.63 0.149 6 Calcium Iilol,gailcl Na phos. (mgldl) (mg/dl) (mE(i/1) (mE(i/1) (m 9.4 8.7 142 4.2 9.8 9..0 141 4.4 9.9 10.1 141 4.0 9.8 10.3 142 4.0 10.2 10.1 143 4.5 9.4 8.9 141 4.2 9.8 0.31 6 9.5 0.72 6 142 4.2 0.8 0.20 6 6 59 A i)iieiiixd 6 tl 7 s Cliiiicalcliemisti-y liidividuavlalues Sex Mal,e Dose level Sample D-1 10 mg/kg Aiiimalskilled oiiscliedule(4 weeks) Animal Nunil)er 3101 3102 3103 :3104 3105 3106 tleaii S.D. N (i')T G[IT r -GTP ALI, Ui-ea Creatiiiiii Glucose T nitrogeii c (it)/1) (IU/I) (IU/I) (IU/I) (mg/dl) (mg/di) (mg/dl) 65 40 66 33 69 32 58 34 67 33 62 31 0 988 12.8 0.5 131 0 755 14.8 0.5 142 0 682 14.9 0.5 145 0 797 11.0 0.4 145 0 615 12.8 0.5 155 0 841 8.1 0.4 132 65 34 0 780 12.4 0.5 142 3.9 3.2 0.0 130.2 2.56 0.05 9.0 6 6 6 6 6 6 6 60 Apl)eiii(xi 6 M - 8 C Iiiiica I clieniisti,@v- Iii(iiv i(luaI va,lties Sex tlale Dose level Saml)le D-1 10 mg/kg Aiiimalskilled on selie(lul(e4 weeks) Study No. BTIR An ima I Ntinibei, 3101 3102 3103 3104 3105 3tO6 tieaii S.D. N Total protein 6.29 6.05 6.28 6-Iii 6.15 6.29 6.20 0.099 6 A Ibtimiii (8/di) A/G ratio 4.15 3.84 4.05 3.94 3.75 3.87 1.94 1.74 1.82 1.77 1.56 1.60 3.93 0.146 6 1.74 0.141 6 Ca Ic itini Iiiorgaiic Na K c plios. (mg/dl) (mg/di) (mEq/1) (mEq/1) (mE 10.2 9.2 143 4.1 1 9.9 10.8 143 4.8 1 9.9 8.8 141 3.9 1 9.7 8.9 142 4.2 1 9.8 9.6 142 4.6 1 9.3 8.5 141 4.3 1 9.8 9.3 142 4.3 1 0.30 0.82 0.9 0.33 1 6 6 6 6 61 Aiiimal Nunil)ei- 4101. 4102 4103 4104 4105 4106 tieaii S.D. N G(IT IL!/I 81 77 76 61 62 83 73 @9.5 6 Ai)i)eiiixd 6 ti 9 si C I iiiica. I cliemis ti,*@8, Iii(iiv i(itiIa va Ities . Sex tlaIe Dose level Sami)le D-1 30 mg/kg Atiiiiiaklislled otiscliedule(4 weeks) (.',IIT G,ri, ALP (Ii-ea Creatiiiin Glucose T iiitrogen cl (IU/I) (IU/I) (IU/I) (mg/dl) (mg/(Il) (mg/di) 46 0 1639 15.1 0.6 139 57 0 1243 15.8 0.5 144 46 0 1036 13.7 0.5 127 47 0 1179 10.4 0.5 140 43 0 940 14.9 0.5 137 78 0 lilt 14.5 0.8 162 53 0 1191 14.1 0.6 142' 13.2 0.0 243.7 1.93 0.12 11.5 6 6 6 6 6 6 62 Animal Number 4101 4102 4103 4104 4105 4106 Mean S.D. N AI)pendix 6 M 10 Clinical cliemistry Individual values Sex Male Dose level Sample D-1 30 mg/kg Animals killed on scliedule(4 weeks) Study No. BM Total protein (&/dl) 6.06 5.87 6.28 5.45 5.87 6.51 6.01 0.368 6 Albumin (8/dl) A/G ratio 3.94 3.97 4.06 3.91 3.86 4.32 1.86 2.09 1.83 2.54 1.92 1.97 4.01 0.166 6 2.04 0.264 6 Calcium (mg/dl) Inorganic phos. (mg/dl) Na (mEq/1) 9.3 8.8 142 9.1 8.3 142 9.9 8.9 143 8.8 9.0 144 9.1 8.7 143 10.2 16.2 147 9.4 10.0 144 0.54 3.06 1.9 6 6 6 K (mEq/1) 4.1 4.1 4.7 3.7 4.3 4.5 4.2 0.35' 6 63 Aptietiidx 6 @l II st Cliiiicalcliemistrv Iii(iivitlualvalues Sex Male Dose level Sample 0-1 0 mg/kg Animals killed on scliedule(Recovery) Animal Number 0107 0108 0109 Otto 0111 0112 tleaii S.D. N GOT (IU/I) 85 77 80 110 76 81 85 12.7 6 GPT (IU/I) 28 30 37 36 40 33 34 4.5 6 r -GTP (IU/I) 0 0 0 0 0 0 0 0.0 6 ALP Urea Creatinin Glucose To nitrogen ell (iti/1) (mg/di) (mg/di) (mg/di) (m 718 22.6 0.5 145 578 1919 0.5 152 621 16.0 0.5 158 542 16.4 0.4 144 652 18.6 0.5 145 564 18.6 0.4 143 613 65.2 6 18.7 2.42 6 0.5 0.05 6 148 5.9 1 6 64 Ai)i)eiiixd 6 @l 12 Cliiiicalcliemistry Iiidividualvalues Sex Male Dose level Sample D-1 0 mg/kg Aiiimalskilled oiiscliedule(Recovery) Study No. BM Aiiimal Number 0107 0108 0109 0110 0111 0112 fleaii S.D. N Total prote@in (8/(Il) Aibumiii (8/dl) 6.43 6.23 6.59 6.65 6.34 6.30 3.82 3.77 3.89 3.110 3.91 3.82 6.42 0.167 6 3.85 0.056 6 A/G ratio 1.46 1.53 1.44 1.42 1.61 1.54 1.50 0.072 6 Calcium liiorgaiiic Na K phos. (mg/di) (mg/di) (mEq/1) (mEq/1) (m 10.0 8.7 143 4.0 97 8;8 142. 4.3 9:8 8.6 140 4.5 9.4 9.0 142 4.2 9.3 8.6 142 3.8 9.4 8.5 139 4.5 9.6 0.28 6 8.7 0.18 6 141 4.2 1.5 0.28 6 6 65 Ai.ii)eixid 6 tl- 13 s Cl iiiicalcliemisti-%, Iiidivi(lualvalues Sex @laIe Dose level Sample U-1 30 mg/kg Animals killed on schedule (Recovery) Animal Number GOT GPT (it)/1) (IU/I) 4107 78 36 4108 86 42 4109 D 4110 74 37 4111 71 44 4112 70 61 Mean 76 44 S.D. 6.5 10.1 N 5 5 D Dead. -R-GTP (IU/I) 0 0 0 0 0 0 0.0 5 ALP (IU/I) 792 852 681 671 586 716 105.4 5 Urea Creatinin Glucose T nitrogen c (mg/dl) (m&/di) (mg/dl) 24.4 0.4 .141 21..7 0.5 171 26.1 0.6 159 19.6 0.5 141 17.1 0.4 150 21.8 3.61 5 0.5 0.08 5 152 12.8 5 66 Aiii)eiiixd 6 M - 14 Clinical cliemistry Individualvalues Sex @iale Dose level Sample D-1 30 mg/kg Animals killed on seliedule(Recovery)' Study No. BM Aiiimal Number Total protein (g/(Il) Albumin (g/dl) 4107 6.08 3.80 4108 6.81 4.13 4109 D 4110 6.70 4.24 4111 6.70 4.25 4112 6.57 4.26 Mean S.D. N D - ead. 6.57 0.288 5 4.14 0.195 5 A/G ratio 1.67 1.54 1.72 1.73 1.84 1.70 0.109 5 Calcium Iiioi,gaiqi Na K phos. (mg/dl) (mg/di) (mEq/1) (mEq/1) (m 9.7 8.3 143 4.1 9.8 9.4 141 9.8 9.4 140 4.1 9.7 9.4 142 4.3 9.6 8.2 142 4.0 9.7 0.08 5 8.9 0.63 5 142 4.2 1.1 0.20 5 5 67 Aiipeiix(i 6 P I s Clinical chemistry Individualvalues Sex Female I)oselevel Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) An ima I Numl)ei- 0201 0202 0203 0204 0205 0206 tleaii S.D. N G(IT (IU/I) 75 67 87 79 88 83 80 8.0 6 GPT (IU/I) 20 26 25 25 23 17 23 3.5 6 r -GTP (IU/I) 0 0 0 1 0 0 0 0.4 6 ALP -Urea Ci-eatinin Glucose T nitrogen c (ILI/1) (mg/dl) (mg/dl) (mg/dl) 364 11.3 0.5 137 431 10-.7 0.6 130 534 14.2 0.5 143 473 12.1 0.5 131 525 12.4 0.5 137 507 10.7 0.5 121 472 11.9 0.5 133 65.2 1.33 0.04 7.6 6 6 6 6 68 Api)etiidx 6 F 2 C Iiiiica I cliemistr,,,, Iii(iividua I va Iues Sex Female [)oselevel Saml)le D-1 0 mg/kg Animals killed on scliedule(4 weeks) Study No. BM Animal Numbei, 0201 0202 0203 0204 0205 0206 Mean S.D. N Total protein (&/dl) 6.52 6.876.63 6.38 6.76 6.62 6.63 0.173 6 A Ibtimiii (8/di) 4.23 @4-24 4.20 4.08 4.15 4.20 4.18 0.060 6 A/G ratio 1.85 .1-61 1.73 1.77 1.59 1.74 1.72 0.099 6 Calcium Itioi,gaici Na K pilos. (mg/di) (mg/dl) (mEq/1) (mEq/1) (m 9.1 7.4 141 4.1 9.5 7.0 143 3.8 9.4 8.0 141 3.7 9.5 9.4 141 4.1 9.4 9.2 141 4.3 9.5 9.5 141 4.4 9.4 0.15 6 8.4 1.09 6 141 4.1 0.8 0.27 6 6 69 Append ix 6 F 3 Stu Cliiiicalcilemisti-y Individualvalues Sex Female Dose level Sample D-1 0.1 mg/kg Animalskilled on schedule(4 weeks) Animal Numbei, 1201 1202 1203 1204 1205 1206 tleail S.D. N G(IT CPT r -GTP ALP Ui-ea Ci-eatiiiinGlucose To nitrogen ch( (ILI/1) (IU/I) (IU/I) (ILI/]) (m&/dl) (mg/di) (m&/di) (m 74 32 64 23 64 25 76 29 71 19 68 22 0 409 .17.0 0.7 126 0 504 10..6 0.5 140 0 362 16.7 0.5 136 0 540 14.9 0.5 182 0 366 9.9 0.5 120 0 331 9.9 0.6 134 70 25 0 419 13.2 0.6 140 5.0 4.8 0.0 84.6 3.41 0.08 22.0 6 6 6 6 6 6 6 70 An ima I Numbei, 1201 1202 1203 1204 1205 1206 Mean S.D. N Append ix - 6 F 4 Clinical chemistry individualvalues Sex Female Dose level Sample D-1 0.1 mg/kg Animals killed on schedule (4 weeks) Study No. Total p I-ote i[I (g/di) Albumin (-O/di) A/G ratio 6.85 6.08 6.61 6.46 6.63 7.00 4.39 3.94 4.19 4.16 4.14 4.30 1.78 1.84 1.73 1.81 1.66 1.59 6.61 0.320 6 4.19 0.154 6 1.74 0.095 6 Calcium (mg/di) Inorganic pilos. (m&/di) Na (mEq/1) 9.9 9.9 141 9.1 9.9 142 10.1 9.9 140 10.8 11.4 142 9.6 9.5 140 9.5 8.5 141 9.8 9.7 141 0.59 1.01 0.9 6 6 6 K (mEq/1) 3.9 4.3 4.4 5.5 4.7 4.0 4.5 0.58 6 71 Animal Number 2201 2202 2203 2204 2205 2206 Mean S.D. N Appendix 6 P 5 Stu Cl iilicalcilemisi.i-yIii(iividtialvalues Sex Female Dose level Sample U-1 I mg/@g Animals killed on scliedule(4 weeks) GOT (;[IT r -GTP ALI' tji-ea Ci-eatiiiiiiGlucose To iiiti-ogen cli (iti/1) (it)/1) (IU/I) (IU/I) (m&/di) (mg/di) (mg/di) (m 62 20 66 16 65 20 67 22 51 23 51 16 0 491 17.7 0.5 156 1 .504 14;1 0.5 148 0 534 13.0 0.4 133 0 377 13.6 0.6 136 0 342 10.6 0.5 142 0 372 9.5 0.5 142 6@O 20 0 437 13.1 0.5 143 7.4 2.9 0.4 82.1 2.89 0.06 8.3 6 6 6 6 6 6 6 72 AI)I)eiiidx 6 F 6 Clinical cliemistry Individualvalues Sex Female Dose level Sample D-1 I mg/kg Animals killed on scliedule-(4weeks) Study No. BMR Animal Number 2201 2202 2203 2204 2205 2206 tleaii S.D. N Total protein (8/dl) 6.48 6.08 6.02 6.86 6.68 6.58 6.45 0.335 6 Albumin (g/di) 4.17 4.11 4.01 4.38 4.11 4.11 4.15 0.125 6 A/G ratio .1.81 2.09 2. 00 1.77 1.60 1.66 1.82 0.191 6 Calcium Inorganic Na K c plios. (mg/dl) (mg/di) (mEq/1) (mEq/1) (mE 9.6 8.5 140 4.4 1 9.9 8.,8 143 5.2 1 9.5 9.3 141 4.4 1 9.8 8.7 140 4.0 1 9.5 8.4 142 3.8 1 9.5 8.7 142 4.4 1 9.6 8.7 141 4.4 1 0.18 0.31 1.2 0.48 1 6 6 6 6 73 Animal Number 3201 3202 3203 3204 3205 3206 Mean S.D. N Ai-)Peiixd 6 F 7 s C Iiiiica I cliemisti-y Iiidiv i(luaI va Ities Sex Female I)oselevel Saml)le U-1 1.0mg/kg Animals killed on schedule (4 weeks) GOT (IU/I) 65 52 66 79 50 46 60 12.5 6 GPT ([U/I) 24 21 27 28 24 20 24 3.2 .6 -r-GTP (IU/I) 0 0 0 0 0 0 0 0.0 6 ALI' (IU/I) 567 361 531 429 382 436 451 81.8 6 Ui,ea Cr@atinin Glucose I nitrogen (mg/di) (mg/di) (mg/dl) 17.4 0.6 122 It.-I 0.5 149 13.7 0.5 '148 14.0 0.5 136 14.7 0.5 126 10.2 0.5 127 13.5 0.5 135 2.59 0.04 11.7 6 6 6 74 Animal Numbei, 3201 3202 3203 3204 3205 3206 Mean S.D. N AI)peiidix 6 F 8 Clinical cliemisti-y Individual values Sex Female Dose level Sample D-1 10 mg/kg Animals killed on scliedule(4 weeks) Study No. B Total protein (S/di) 6.68 6.84 7.14 6.58 7.21 7.03 6.91. 0.255 6 Albumin (8/di) 4.35 4.34 4.52 4.24 4.64 4.40 4.42 0.143 6 A/G ratio 1.87 1.74 1.73 1.81 1.81 1.67 1.77 0.072 6 Calcium (m&/dl) 9.6 9.2 10.2 9.5 9.9 9.5 9.7 0.35 6 Inorganic phos. (m&/dl) Na (mEq/1) 7.0 141 7.6 140 7.9 143 8.0 141 9.2 144 7.6 142 7.9 142 0.73 1.5 6 6 K (mEq/1) 3.7 3.9 4.8 4.5 3.7 4.2 4.1 0.45 6 75 Animal Number 4201 4202 4203 4204 4205 4206 Mean S.D. N Ai)ijeiiix(i 6 p 9 st iiiicaI cliemisti,y Iiidiv itiuaI va Ities Sex Female Dose level Sample D-1 30 mg/kg Animals killed on selie(lul(e4 weeks) GOT (IU/I) 62 55 57 55 50 59 56 4.1 6 G[IT (IU/I) 40 31 31 36 29 48 36 7.2 6 r -GTP (IU/I) 0 0 0 0 0 0 0 0.0 6 ALI' ([U/I) 826 609 593 717 404 392 590 170.9 6 Urea Creatiniii Glucose To nitrogen ch (mg./di) (mg/di) (mg/dl) (m 14.3 0.6 152 20.9 0.6 125 19.8 0.5 111 13.7 0.5 119 21.8 0.5 116 17.3 0.5 135 18.0 3.43 6 0.5 0.05 6 126 15.0 6 76 41)1)eiid F 10 Cl iiiicalcliemisti,,vlildivi(itviaallues Sex Female Dose level saml)le D-1 30 mg/kg Animals killed otiscliedule(4 weeks) Study No. BM Aiiima I Ntiml.ier Total proteiii (8/di) A Ibtimi11 (8/di) 4201 4202 4203 4204 4205 4206 7.17 7.32 6.88 6.64 6.67 7.39 4.61 4.68 4.53 4.36 4.56 4-.77 tleaii S.D. N 7.01 0.327 6 4.59 0.140 6 4/(-. I*atio 1.80 1.77 1.93 1.91 2.16 1.82 1.1.10 0.143 Calcium Iiiol.gailcl Na K 1)[10S. (nig/di) (mg/di) (PiEq/1) (mEq/1) (m 9.5 7.0 143 4.4 10.0 8;.9 142 4.7 9.9 .9.1 142 4.2 9.7 9.3 140 5.5 (1).5 7.9 143 4.4 10.8 9.3 142 6.0 9.9 0.49 6 8.6 0.93 6 142 .1.1 6 4.9 0.72 6 77 Al)l)eiiid 6 F - 11 .st Cl iiiicalclieniistr-,\Ipiidivi(ILvialues Sex Female I)ose level Sample D-1 0 mg/kg Aiiimals killed oiisclie(lule(Recovet-Y) Aiiimal Nii n1e-.ii, 0207 0208 0209 0210 0211 0212 tleaii S.D. N (II)T (IU/I) -)7 7.5 71 66 78 81 71 8.8 6 (;PT (IU/I) 24 30 22 28 29 44 30 7.7 6 (IU/I) 0 0 0 0 0 0 0 0.0 6 ALI' I(I/ I 266 273 411 283 393 313 323 63.4 6 Llt*ea Ci-eatiiiiiiGlucose T iiitrogen c (nig/dI (mg/di) (mg/dl) 18.0 0.6 152 19.9 0.6 154 15.0 0.6 149 20.6 0.6 141 14.9 0.5 144 15.5 0.5 160 17.3 0.6 150 2.55 0.05 6.9 6 6 6 78 Aljl)eiixi(i 6 p 12 C I iiiica I clienisti,@\, Iii(iiv idua I va Iues Se.\ Female [)oselevel Sami)le D-1 0 mg/kg Animals killed on scliedule(Recovery) Study No. BM Animal Ntimliei, Total i)roteiii (8/(Il) Ail)tjmin A/G i*atio 0207 0208 0209 0210 0211 0212 tleail S.D. N 7.00 7.26 7.01 7.18 6.47 7.17 7.02 0.286 6 4.35 4.41 4.42 4.38 4.05 4.40 4.34 0.142 6 1.64 1.55 1.71 1.56 1.67 1.59 1.(i2 0.064 6 Calcium liioi-ganic Na pilos. (mg/dl) (ms/di) (mEq/1) (mEq/1) (m 9.8 7.,l 140 4.1 9.7 6;7 140 4.0 9.5 7.9 141 4.2 9.6 6.8 140 4.2 9.4 7.9 140 4.0 9.8 7.6 139 3.9 9.6 0.16 6 7.3 0.54 6 140 4.1 0.6 0.12 6 6 79 Al)lieiiixd 6 P 13 st C Iiiiica I cliepiisti-,v Iiidiv idua I va Ities Sex Fema Ie [)oselevel Saml)le D-1 30 mg/kg Animals killed on scliedule(Recovery) knimal Nunil)Ie- 4207 4208 4209 4210 4211 4212 Meal) S.D. N G(I'T (;PT ([(!/I) (ILI/1) (i2 41 53 29 47 25 78 25 64 28 62 31 61 30 10.6 5.9 6 -GTP (IU/I) 0 0 0 0 0 0 0 0.0 6 ALI' (IL)/I) Ui-ea nitrogen (mg/di) 404 29.2 347 22..5 363 22.6 486 20.2 353 22.2 369 21.7 387 52.4 24.1 2.99 6 CreatiiiiiiGlucose T c (mg/di) (mg/di) 0.6 132 0.6 141 0.5 123 0.6 117 0.6 116 0.6 .129 0.6 126 0.04 9.6 6 6 80 AI)peiiidx 6 F 14 Cl iiiicalcliemisti,,vIndividual values Sex Female I)oselevel sample 0-1 30 mg/kg Aiiimalskilled on scliedule(Recovery) Study No. BM Animal -Numbei, 4207 4208 4209 4210 4211 4212 Mean S.D. N Total 1)rote iii (8/(Il) A I1)(Iinii (8/di) 7.63 7.51 7.24 8.18 7.12 7.34 4.72 4.85 4.55 4..99 4.43 4.79 7.50 0.379 6 4.72 0.204 A/(; i-at io 1.62 1.82 1.69 1.56 1.65 1.88 1.70 0.123 6 Calcium Iiioi*gaici Na I)IIOS. (mg/di) (mg/di) (mEq/1) (mEq/1) (m 10.2 .8.2 141 4.7 10.4 7@.9 141 3.9 10.4 7.7 141 4.1 10.3 8.1 141 3.9 9.9 8.6 141 4.2 10.1, 8.2 142 4.8 10.2 0.19 6 8.1 0.31 6 141 4.3 0.4 0.39 6 6 Animal Number 0101 0102 0103 0104 0105 0106 0107 0108 0109 0110 0111 0112 Appendix 7M 1 Ui-inalysis Individual values Sex Male Dose level Sample D-1 0 mg/kg 3 weeks after commencement of treatment Study No. Ui,obilinogen (EU/dl) Occult Blood 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.11 0.1 0.1 0.1 - ... Trace Ti-ace Bilirubin Ketone bodies (mg/dl) - 5 - 5 - 15 - 5 - 5 - 5 - 5 - 5 - - - 5 - 5 - 5 Negative, + Slight, Moderate, Glucose (g/di) - Severe Protein (mg/dl) 30 30 30 30 30 Trace 30 30 30 30 Trace 30 82 Animal Number 1101 1102 1103 1104 1105 1106 Appendix 7M2 Urinalysis Individual values Sex Male Dose level Sample D-1 0.1 3 weeks after commencement of treatment mg/kg Study No. Urobilinogen (EU/dl) Occult Blood Bilirubin Ketone bodies (mg/dl) 0.1 - 5 0.1 - 5 0.1 - 15 0.1 Trace - 5 0.1 - 40 0.1 - 5 Negative, + Slight, Moderate, Glucose (g/di) - Severe Protein (mg/dl) 30 30 30 30 30 100 83 Animal Number 2101 2102 2103 2104 2105 .2106 Appendix 7M3 Urinalysis Individual values Sex Male Dose level Sample D-1 I mg/kg 3 weeks after commencement of treatment Study No. Urobilinogen (EU/dl) Occult Blood Bilirubin Ketone bodies (mg/dl) Glucose (g/dl) 0.1 - - 15 0.1 - - 5 0.1 - - 5 0.1 - - 5 0.1 Ti'ace - 5 0.1 - 5 Negative, + Slight, Moderate, +++ Severe Protein (mg/dl) 30 30 30 100 100 30 84 Animal Number 3101 3102 3103 3104 3105 3106 Appendix 7M 4 Urinalysis Individual values Sex Male Dose level Sample D-1 10 mg/kg 3 weeks after commengement of treatment Study No. Urobilinogen (EU/dl) 0.1 0.1 0.1 0.1 0.1 0.1 Occult Blood - Bilirubin Ketone bodies (mg/dl) 5 - 5 - 5 - 5 - 5 - 5 Glucose (g/dl) - Negative, Slight, Moderate, +++ Severe Protein (mg/dl) 30 30 100 30 100 30 85 Appendix 7M 5 Urinalysis Individual values Sex Male Dose level Sample D-1 30 mg/kg 3 weeks after commencement of treatment Study No. Animal Number 4101 4102 4103 4104 - 4105 4106 4107 4108 4109 4110 4111 4112 Urobillnogen (EU/dl) 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 6.1 0.1 0.1 Occult Blood - Bilirubin Ketone bodies (mg/dl) - 15 - 5 - 40 - 5 - 5 - 5 - 5 - 5 - 15 5 40 5 Negative, Slight, ++ Moderate, Glucose (g/dl) Severe Protein (mg/di) 30 100 30 30 30 Trace 30 30 30 30 100 30 86 Animal Number 0107 0108 0109 0110 0111 0112 Appendix 7 M.- 6 Study No. B Urinalysis Individual values Sex Male Dose level Sample D-1 0 mg/kg 6 weeks after commencement of treatment (Recovery) Urobillnogen (EU/dl) Occult Blood Billrubin Ketone bodies (mg/dl) Glucose (g/dl) Oll 0.1 Trace 0.1 Trace 0.1 0.1 Trace 0.1 Trace 6 - 5 - - 5 - 5 - 5 - Negative, + Slight, Moderate, +++ Severe Protein p (mg/dl) -Trace i 30 A 30 f Trace I 30 30 87 Animal Number 4107 4108 4109 4110 4111 4112 Appendix 7M7 'Study No. Urinalysis Individual values Sex Male Dose level Sample D-1 30 mg/kg 5 weeks after commencement of treatment (Recovery) Urobilinogen (EU/dl) Occult Blood Bilirubin Ketone bodies (mg/dl) Glucose (g/dl) 0.1 5 0.1 ++ - 5 D 0.1 7 - 5 0.11 - - 5 0.1 - 5 Negative, D Dead. Slight,, ++ Moderate, +++ Severe Protein (mg/dl) 30 30 30 30 30 88 Animal Number 0201 0202 0203 0204 0205 .0206 0207 0208 0209 0210 0211 0212 Appendix F Urinalysis Individual values Sex Female Dose level Sample D-1 0 mg/kg 3 weeks after commencement of treatment Study No. Urobillnogen (EU/dl) Occult Blood 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Trace 0.1 0.1 0.1 Trace 13ilii,ubin Ketone bodies (mg/dl) 5 - 5 5 - Glucose .(g/dl) - Negative, + Slight, Moderate, +++ Severe Protein (mg/dl) 30 30 30 30 30 30 30 30 Trace 30 30 Trace 89 Animal Number 1201 1202 1203 1204 1205 1206 Appendix 7F2 Urinalysis Individual value.s Sex Female Dose level Sample D-1 .0.1 mg/kg 3 weeks after commencement of treatment Study No. Urobilinogen (EU/dl) Occult Blood o@. I Ti-ace 0.1 0.1 0.1 Trace 0.1 0.1 Billrubin Ketone bodies (mg/dl) 5 Glucose (g/dl) - Negative, + Slight, ++ Moderate, +++ Severe Protein (mg/dl) .30 Ti*ace 30 30 Trace 30 90 Animal Number 2201 2202 2203 2204 2205 2206 Appendix 7 F -.3 Urinalysis -Individual values Sex Female Dose level Sample D-1 I mg/kg 3-weeks after commencement of treatment Study No. Urobilinogen (EU/dl) Occult Blood B ilirubin Ketone bodies (mg/dl) 0.1 0.1 Trace 0.1 Trace 0.1 0.1 + 0'.1 + 5 Negative, + Slight, Moderate, Glucose (g/dI.) Severe Protein (mg/dl) 30 30 Trace 30 30 30 Animal Number 3201 3202 3203 3204 3206 3206 Appendix 7F 4 Urinalysis Individual values . Sex Female Dose level Sample D-1 10 mg/kg 3 weeks after commencement of treatment Study No. Urobillnogen (EU/di) Occult Blood 0.1 0.1 0.1 0.1 O..l 0.1 Trace Billrubin Ketone bodies (mg/dl) 5 - 5 Glucose (g/dl) - Negative, + Slight, Moderate, +++ Severe Protein (mg/dl) 30 Trace Trace 30 30 30 92 Animal Number 4201 4202 4203 -4204 4205 4206 4@-07 4208 4209 4210 4211 4212 Appendix 7 F 5 Urinalysis Individual values Sex Female Dose level Sample D-1 3 weeks after commencement of 30 treatment mg/kg Study No. Ui,obilinogen (EU/dl) Occult Blood 0.1 + 0.1 - 0.1 - 0.1 - 0.1 - 0.1 Trace 0.1 Trace 0.1 + Oll 0.1 0.1 0.1 Trace Billrubin Ketone bodies (mg/dl) Glucose (g/di) - 5 - Negati ve, + Slight, Moderate, +++ Severe Protein (mg/dl) Trace Trace 30 - 30 30 Trace Trace 30 Trace 93 Animal Number 0207 0208 0209 0210 0211 0212 Appendix 7 F -6 Urinalysis Individual values Study No. Sex Female Dose level Sample D-1 0 mg/kg .5 weeks after commencement of treatment (Recovery) Urobillnogen (EU/dl) Occult Blood Bilirubin Ketone bodies (mg/dl) 0.1 Trace 0.1 0.1 0.1 0.1 0.1 Trace Negative, + Slight, Moderate, Glucose (g/dl) Severe Protein (mg/dl) Trace 30 Trace Trace 30 94 Animal Number 4207 4208 4209 4210 4211 4212 Appendix 7F7 Urinalys is . Individual valu es Study No. B@ . Sex Female Dose level Sample D-1 30 mg/kg 6 weeks after commencement of treatment (Recovery) Urobillnogen (EU/dl) Occult Blood Bllirubin Ketone bodies (mg/dl) 0.1 0.1 0.1 0.1 0.1 O..l, Ti,ace Trace Trace Trace Negative, + SI.Ight, Modei-ate, Glucose (g/di) - Severe Protein pt (mg/dl) 30 .6. Trace 7. Trace 7. 30 7. Trace 8. Trace 7. 95 Appendix am 1 Organ weight (Absolute) Individual values Sex Male Dose level Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) Study No. B Animal Number F.B.W (g) Brain (g) 0101 354 0102. 318 0103 334 0104 362 0105 377 0106 370 1.90 2.00 1.77 1.95 2.03 2.11 Mean S.D. N F.B.W 351 22.0 1.96 0.117 6 Final body weight Liver (g) 13.07 12.17 13.50 12.92 14.62 15.42 13.60 1.180 6 Kidneys (g) 2.38 2.16 2.45 2.46 2.50 2.63 2.43 0.156 6 Adrenals (mg) 51.2 53.1 47.6 64.4 46.1 50.4 50.5 3.19 6. Testes (g) 3.01 3.23 2.54 3.07 3.24 2.84 2.99 0.265 6 96. Appendix 8 M'- 2 Organ weight (Absolute) Individua:l values Sex Male Dose level Sample D-1 0.1 mg/kg Animals killed on schedule (4 weeks) Study No. Animal Number F.B.W (g) Brain (g) 1101 1102 1103 1104 1105 1106 375 2.06 321 2.13 362 1.90 321 1.94 376 1.92 399 1.92 Mean S.D. N F.B.W 359 31.8 6 1.98 0.094 6 Final body weight Liver (g) 16.05 13.25 13.55 13.52 16.24 15.37 14.50 1.194 6 Kidneys (g) 2.76 2.19 2.60 2.27 2.71 2.78 2.55 0.257 6 Adrenals (mg) 52.2 55.6 60.4 63.4 53.9 50.3 64.3 4.90 6 Testes (g) 2.92 2.92 3.20 3.18 3.17 3.44 3.14 0.196 6 97 Appendix 8m 3 Organ weight (Absolute) Individual values Sex Male Dose level Sample D-1 1 mg/kg Animals killed on schedule (4 weeks) Study No. Animal Number F.B.W (g) Bi"ain (g) 2101 2102 2103 2104 2105 2106 .346 345 354 321 376 350 1.99 1.89 1.95 2.00 1.91 1.92 Mean S.D. N .F.B.W 349 17.7 6 1.94 0.045 6 Final body weight - Liver (g) 14.63 12.98 14.84 12.15 15.90 12.75 13.88 1.459 6 Kidneys (g) 2.42 2.24 2.46 2.26 2.18 1.99 2.26 0.170 6 Adrenals (mg) 60.6 47.1 57.4 52.4 55.3 64.8 52.9 3.71 6 Testes (g) 2.85 3.10 3.04 3.15 3.40 3.30 3.14 0.194 6 98 Appendix 8M4 Organ weight (Absolute) Sex Mate Dose level Sample Animals killed on schedule Individual values D-1 10 mg/kg (4 weeks) Study No. Animal Number F.B.W (g) Brain (g) 3101 3102 3103 3104 3105 3106 326 1.83 341 1.92 320 1.96 335 2.07 389 1.95 345 1.92 Mean s.D. N F.B.W 343 24.5 6 1.94 0.078 6 Final body weight Liver (g) 17.23 18.33 17-38 17.50 15.01 16.10 16.93 1.179 6 Kidneys (g) 2.46 2.28 2.35 2.56 2.33 2.37 2.39 0.101 6 Adrenals (mg) 46.3 46.6 52.2 47.6 68.8 45.5 49.5 5.14 6 Testes (g) 2.78 2.84 3.23 3.11 3.18 2.91 .03..01 189 6 99 Appendix 8 m B. Organ weight (Absolute) Individual values Sex Male Dose level Sample D-1 30 mg/kg Animals killed on schedule (4 weeks) Study No. Animal Number F.B.W (g) Brain (g) 4101 4102 4103 4104 4105 4106 300 1.92 286 2.11 321 1.97 283 1.97 269 1.92 293 1.93 Mean S.D. N F.B.W 292 17.6 6 1.97 0.072 6 Final body weigh.t Liver (9) 19.22 20.73 24.20 18.08 18.72 21.22 20.36 2.228 6 Kidneys (g) 2.02 2.16 2.'21 2.04 2.03 2.-17 2.11 0.084 6 Adrenals. (mg@) 44.3 48.'5 54 0 49:4 50.3 56.0 50.4 4.16 6 Testes (g) 2.62 3.30 3.26 j.-03 3.21 3.06 3.08 0.250 6 100 Appendix 8M6 Organ weight (Absolute) Sex Ma.1 e Dose l@vel Sample Animals killed on schedule Individual.values-. D-1 0 mg/kg (Recovery) Study No. Animal Number F.B.W (g) Brain (g) 0107 0108. 0109 0110 0111 0112 365 2.07 405 1.98 438 2.10 374 2.01 344 2.04 410 2.11 Mean S.D. N F.B.W 389 34.4 6 2.05 0.051 6 Final body weight Liver (g) 12.43 13.98 15.91 12.82 11.18 15.12 13.57 1.768 6 Kidneys (g) 2.29 2.35 2.50 2.62 2.41 2.67 2.47 0.151 6 Adrenals (mg) 48.5 64.0 46.9 59.0 46,.8 63.1 53.11 6.85 .6 Testes (g) 3.00 3.36 2.89 3.24 0.86 3.12 2.74 0.942 6 1.01 Appendix 8 M-7 Organ weight (Absolute) Individual* values Sex Male Dose level Sample D-1 30 mg/kg Animals killed on schedule (Recovery) Study No. Animal Number F.B.W (g) Brain (g) 4107 4108 4109 4110 4111 4112 326 336 D 331 340 373 1.96 2.09 2.05 1.96 2.04 Mean S.D. N 341 18.5 2.02 0.058 F.D.W Final body weight D.: Dead. Liver (g) 17.69 21-36 21.91 20.99 21.35 20.66 1.693 Kidneys (g) 2 .23 2.30 2.33 2.56 2.57 2.40 0.157 Adrenals (mg) 53.4 61.5 53.9 .45.0 43.6 51.5 7.31 Testes (g) 3.26 3.40 3.54 3.43 3.27 3.38 0.117 102 Appendix 8FI Organ weight (Absolute) Individual values Sex Female Dose level Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) Study No. E Animal Number F.B.W (g) Brain (g) 0201 228 1.87 0202- 214 1.78 0203 221 1.86 0204 264 1.96 0205 227 1.84 0206 227 1.78 Mean S.D. N F.B.W 230 17.4 6 1.85 0.067 6 Final body weight Liver (g) 8.53 8.75 8.93 9.60 8.4-0 8.32 8.76 0.471 6 Kidneys. (g) 1.59 1.36 1.62 1.88 1.72 1.52 1.62 0.177 6 Adrenals (mg) 50.8 58.4 66.6 63.4 59.1 62.5 60.1 5.46 6 Ovaries (mg) 95.2 89.2 76.7 93.7 72.7 75.6 83.7 10.13 6 103 Appendix 8F 2 Organ weight (Absolute) Individual values Sex Female' Dose level Sample D-1 0.1 mg/kg Animals killed on schedule (4 weeks) Study No. Animal Number F.B.W (g) Brain (g) 1201 1202 1203 1204 1205 1206 224 1.83 212 1.85 235 1.81 242 1.72 227 1.81 ?22 -1.73 Mean S.D. N F.B.W 227 10.5 6 1.79 0.054 6 Final body weight Liver (g) 8.57 7.83 9.62 9.41 8.38 8.18 8.65 0.679 6 Kidneys (g) 1.49 1.42 1.63 1.89 1.65 1.60 1.61 0.162 6 Adrenals (mg) 60.5 50.3 55.8 60.9 71.3 60.1 59.8 6.93 6 Ovaries (mg) 78.0 79.7 93.0 85.5 92.1 73.7 83.7 7.86 6 104 Appendix 8-F-3 Organ weight (Absolute) Individual values Sex Female Dose level Sample D-1 I mg/kg Animals killed on schedule (4 weeksl Animal Number F.B.W (g) Brain (g) 2201 225 2202 226 2203 216 2204 240 2205 259 2206 246 1.87 1.92 1.81 2.02 1.90 1.91 Mean S.D. N F.B.W 235 151.9 6 1.91 0.069 6 Final body weight Liver (g) 9.55 8.39 7.95 9.33 10.32 8.85 9.07 0.852 6 Kidneys (g) 1.72 1.71 1.57 1.62 1.79 1.63 1.67 0.081 6 Adrenals (mg) 48.1 66.5 59.1 66.8 68.3 66.5 -62.6 7.79 6 Study No.. Bb Ovaries (mg) 7B.9 93.7 93.8 116.1 75.6 .96.8 92.0 15.11 6 105 Appendix 8F4 Organ weight (Absolute) Individua,l values Sex Female Dose level Sample D-1 10 mg/kg ,Animals killed on schedule (4 weeks) Study No. Animal Number F.B.W (g) Brain (g) 3201 3202 3203 3204 3205 3206 216 1.96 225 1.83 186 1.77 221 1.86 212 1.84 223 1.82 Mean s.D. N F.B.W 214 14.4 6 1.85 0.063 6 Final body weight Liver (g) 10.04 11.96 9.06 10.75 11.01 10.68 10.58 0.973 6 Kidneys (g) 1.69 1.81 1.51 1.76 1.64 1.76 1.70 0.109 6 Adrenals (mg) 62.6 71.4 .48.4 55.0 63.9 63.7 60.8 8.01 6 Ovaries (mg) 90.1 72.6 63.7 76.8 74.7 76.7 75.8 8.53 6 106 Appendix 8F 5 organ weight (Absolute) Sex Dose level Female Sample Animals killed on schedule Individual D-1 (4 weeks) values 30 mg/kg Study No Animal Number F.B.W (g) Brain (g) 4201 4202. 4203 4204 4205 4206 178 1.79 194 1.80 194 1.80 204 1.96 181 1.81 3.91 1.82 Mean S.D. N F.B.W 190 9.5 6 1.@83 0.064 6 Final body weight Liver (g) 12.25 14.65 14.18 14.04 13.81 13.73 13.78 0.816 6 Kidneys (g) 1.28 1.46 1.51 1.62 1.53 1.62 1.50 0.126 6 Adrenals (mg) 55.9 60.0 55.5 64.6 47.0 50.8 65.6 6.28 6 Ovaries (mg) 66.0 61.6 76.2 56.4 66.7 58.6 62.6 7.57 6 107 Appendix 8F 6 Organ weight (Absolute) Individual values Sex Female Dose level Sample D-1 0 mg/kg Animals killed on schedule (Recovery) Study No. Animal Number F.B.W (g) Brain (g) 0207 0208 0209 0210 0211 0212 325 266 264 270. 278 257 1.91 1.85 1.93 2.04 1.99 1.86 Mean S.D. N F.B.W 277 24.8 6 1.9 3 0.074 6 Final body weight Liver (g) 11.30 8.12 8.55 8.97 8.81 8.80 9.09 1.122 6 Kidneys (g) 1.92 1.63 1.72 1.84 1.78 1.76 1.78 0.100 6 Adrenals (mg) 80.7 61.1 60.3 82.7 77.7 72.0 70.8 12.56 6 Ovaries (mg) 106.4 86.3 78.3 96.7 83.6 88.-l 89.7 9.96 6 108 Appendix 8-F 7 Organ weight (Absolute).- Individual values Sex Female , Dose level Sample D-1 30 mg/kg Animals killed on schedule (Recovery) Study No. Animal Number F.B.W (g) Brain (g) 4207 4208 4209 4210 4211 4212 .201 199 208 193 213 234 1.71 1.72 1.90 1.90 1.89 2.06 Mean S.D. N F.B.W 208 14.6 6 1.86 0.131 6 Final body weight Liver (g) 11.92 13.04 12.90 11.64 12.40 12.85 12.46 0.674 6 Kidneys (g) 1.44 1.62 1.57 1.47 1.57 1.85 1.59 0.146 6 Adrenals (mg) 50.1 53.7 46.7 53.0 53.6 67.7 54.0 7.39 6 Ovaries (mg) 61.8 59.3 72.7 73.6 71.2 72.1 68.5 6.22 6 109 Appendix 9m I Study No. Organ weight (Relative - Individual values percentage of body Sex Male Dose level Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) weight) Animal Number F.B.W (g) -Brain Liver Kidneys Adrenals (Xlo-3) Testes 0101 0102 oioi 0104 0105 0106 354 0.54 318 0.63 334 0.53 352 0.55 377 0.54 370 0.57, Mean S.D. N F.B.W 351 22.0 6 0.56 0.037 6 Final body weight 3.69 3.83 4.04 3.67 3.85 4.17 3.88 0.197 6 0.67 0. 68 0..73 0.70 0.66 0.71 0.69 0.026 6 14.5 16.7 14.2 15.5 12.2 13.6 14.5 1.65 6 0.85 1.02 0.76 0.87 0.86 0.77 0.86 0.094 6 110 Appendix 9M 2 Study No. Organ weight (Relative Individual values percentage of body weight) Sex Male Dose level Sample D-1 0.1 mg/kg Animals killed on schedule (4 weeks) Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (XIO-3) Testes 1101 1102 1103 1104 1106 1106 375 0.55 321 0.66 362 0.52 321 0.60 376 0.51 399 0.48 Mean S.D. N F.B.W 359 31.8 6 0.55 0.066 6 Final body weight 4.28 4.13 3.74 4.21 4.05 3.85 4.04 0.210 6 0.73 0.68 0.72 0.71 0.72 0.70 0.71 0.018 6 13.9 17.3 13.9 19.8 14.3 12.6 15.3 2.70 6 0.78 0.91 0.88 0.99 0.84 0.86 0.88 0.071 6 Appendix 9M3 Study Organ weight (Relative Individual values percentage of body Sex Male Dose level Sample D-1 I mg/kg Animals killed on schedule (4 weeks) weight) No.. Animal Number F.B.W (g) Brain 2101 2)02. 2103 2104 2105 2106 346 345354 321 376 350 0.58 0.55 0.55 0..62 0.51 0.55 Mean S.D. N F.B.W 349 17.7 6 0.56 0.037 6 Final body weight Liver 4.23 3.76 4.19 3.79 4.23 3.64 3.97 0.272 6 Kidneys 0.70 0.65 0.69 0.70 0.58 0.57 0.65 0.060 6 Adrenals (XIO-3) 14.6 13.7 16.2 16.3 14.7 15.7 15.2 1.03 6 Testes 0.82 0.90 0.86 0.98 0.90 0.94 0.90 0.057 6 112 Appendix 9M4 Study No. Organ weight (Relative.: percentage of Individual*values Sex Male Dose level Sample D-1 10 Animals killed on schedule (4 weeks) body weight) mg/kg Animal Number F.B.W (g) Brain 3101 3102 3103. 3104 3105 3106 326 0.56 341 0.56 3.20 0.61 335 0.62 389 0.50 345 0.56 Mean S.D. N F.B.W 343 24.5 6 0.57 0.043 6 Final body weight Liver 5.29 6.38 5.43 5.22 3.86 4.67 4.98 0.611 6 Kidneys 0.75 0.67 0.73 0.76 0.60 0.69 0.70 0.060 .6 Adrenals (XIO-3) 14.2 13.7 16.3 14.2 15.1 13.2 14.5 1.10 6 Testes 0.85 0. 83 1.01 0.93 0.82 0.84 0.88 0.075 6 '113 Appendix 9M 5 Study No. Organ weight (Relative Individual values percentage of Sex Dose level Male Sample D-1 30 Animals killed on'schedule (4 weeks) body weight) mg/kg Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (XIO-3 Testes 4101 4102 4103 4104 -4105 4106 300 0.64 286 0.74 321 0.61 283 0.70 269 '0.71 293 0.66 .Mean S.D. N F.B.W 292 17.6 6 0.68 0.048 6 Final body weight 6.41 7.25 7.54 6.@9 6.96 7.24 6.97 0.475 6 0.67 0.76 0.69 0.72 0.76 0.74 0.72 0.035 6 14.8 17.0 16.8 17.5 18.7 19.1 17.3 1.54 6 0.87 1.15 1.02 1.07 1.19 1.04 1.06 0.112 6 114 Appendix 9 M-6 Study No. Organ weight (Relative percentage of body Individual values Sex Male Dose level Sample D-1 0 mg/kg Animals killed on schedule (Recovery) weight) Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (XIO-3 Testes 0107 0108 0109 0110 0111 0112 365 405 438* 374 344 410 0.57 0.49 0.48 0.54 0.59 0.51 3.41 3.45 3.63 3.43 3.25 .3-69 0.63 0.58 0.5-7 0.70 0.70 0.65 13.3 13.3 10.7 15.8 13.6 16.4 0.82 0.83 0 66 0:87 0.25 0.76 Mean. s.D. N F.B.W 389 34.4 6 0.53 0.044 6 Final body weight 3.48 0.160 6 0.64 0.056 6 13.7 1.82 6 0.70 0.231 6 115 Appendix 9M 7 Study No. Organ weight (Relative Individual values percentage of Sex Dose level Male Sample D-1 30 Animals killed on schedule (Recovery) body weight) mg/kg Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (XIO-3 Testes 4107 4108 4109 4110 4111 4112 326 336. D 331 340 373 0.60 0.62 0.62 0.58 0.56 Mean S.D. N 341 18.5 5 F.B.W !Final body D : Dead. 0.59 0.030 5 -weight @5.43 6.36 6.62 6.17 5.72- 6.06 0.482 5 0.68 0.68 0.70 0.75 0.69 0.70 0.029 5 16.4 18.3 16.3 13.2 11.7 15.2 2.67 5 1.00 1.01 1.07 1.01 0.88 0.99 0.069 5 116 Appendix 9F1 Study No. Organ weight (Relative Individual values percentage of body Sex Female Dose level Sample D-1 0 mg/kg Animals killed on schedule (4 weeks) weight) Animal Number F.B.W (g) Brain Liver Kidneys Adi*e.nals (XIO-3) Ovaries (Xlo-3) 0201 0202@ 0203 0204 0205 0206 228 214 221 @264 227 227 0.82 0.83 0.84 0.74 0.81 0.78 3.74 4.09 4.04 3.64 3.70 3.67 0.70 0.64 0.73 0.71 0.76 0.67 22.3 .27.3 30.1 24.0 26.0 27.5 41.8 41.7 34.3 35.5 32.0 33.3 Mean S.D. N F.B.W 230 17.4 6 0.80 0.037 6 Final body weight 3.81 0.198 6 0.70 0.043 6 26.2 2.76 6 36.4 4.26 6 117 Appendix 9F2 Study No. Organ weight (Relative Individual values percentage of body weight) Sex Female Dose level Sample D-1 0.1 mg/kg Animals killed on schedule (4 weeks.) Animal Number F.B.W (g) Brain 1201 1202 1203 1204 1205 1206 224 0.82 212 0.87 235 0.77 242 0.71 227 0.80 222 0.78 Mean S.D. N F.B.W 227 10.5 6 0.79 0.053 6 Final body weight Liver 3.83 3.69 4.05 3.89 3.69 3.68 3.81 0.148 6 Kidneys 0.67 0.67 0.'69 0.78 0.73 0.72 0.71 0.042 6 Adrenals (XIO-3) 27.0 23.7 23.7 25.2 31.4 27.1 26.4 2.89 6 Ovaries (Xlo-3) 34.8 37.6 39.6 35.3 40.6 33.2 36.8 2.88 6 Appendix 9F 3 Study No. Organ weight (Relative Individual values percentage of body Sex Female Dose level Sample D-1 1 mg/kg Animals killed on schedule (4 weeks) weight) Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (Xlo-3 Ovaries (XJO-3 2201 2202 2203 2204 2205 2206 225 0.83 226 0.85 216 0.84 240 0.84 259 0.73 246 0.78 Mean S.D. N F.B.W 236 15.9 6 0.81 0.047 6 Final body weight 4.24 3.71 3.68 3.89 3.98 3.60 3.85 0.237 6 0.76 0.76 0.7@ 0.68 0.69 0.66 0.71 0.043 6 21.4 29.4 27.4 27.8 26.,4 27.0 26.6 .2.73 6 33.7 41.5 43.4 48.4 29.2 39.3 39.3 6.90 6 119 Appendix 9F4 Study No. B Organ weight (Relative percentage of - Individual values Sex Female Dose level Sample D-1 10 .Animals killed on schedule,(4 weeks) body weight) mg/kg Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (Xlo-3 Ovaries (Xlo-3 3201 3202, 3203 3204 3205 3206 216 0.91 225 0.81 186 0.96 221 0.84 212 0.87 223 0.82 Mean S.D. N F.B.W 214 14.4 6 0.87 0.055 6 Final body weight 4.65 5.32 4.87 4.86 5.19 4.79 4.95 0.255 6 0.78 0.80 0.81 0.80 0.77 0.79 0.79 0.015 6 29.0 31.7 26.0 24.9 30.1 28.6 28.4 2.54 6 41.7 32.3 34.2 34.8 35.2 34.4 35.4 3.23 6 120 Animal Number F.B.W (g) Appendix 9F5 Study No. Organ weight (Relative percentage of - Individual values Sex Female Dose level Sample D-1 30 Animals killed on schedule (4 weeks) body weight) mg/kg Brain Liver Kidneys Adrenals (Xlo-3) Ovaries (Xlo-3) 4201 4202. 4203 4204 4205 4206 178 1.01 194 0.93 194 0.93 204 0.96 181 1.00 191 0.95 Mean S.D. N F.B.W 190 -9.5 6 0.96 0.034 6 Final body weight 6.88 7.55 7.31 6.88 7.63 7.19 7.24 0.321 6 0.72 0.75 0.78 0.79 0.85 0.86 0.79 0.053 6 31.4 30.9 28.6 31.7 26.0 26.6 29.2 2.50 6 37.1 31.8 39.3 27.6 31.3 30.7 33.0 4.37 6 Appendix 9F6 Study No. Organ weight (Relative percentage of body Individual values Sex Female Dose level Sample D-1 0 mg/kg Animals killed on schedule (Recovery) weight) Animal Number F.B.W (g) Brain Liver Kidneys Adrenals (XIO-3) Ovaries (Xlo-3 0207 0208. 0209 0210 0211 0212 325 0.59 3.48 0.59 24.8 32.7 265 0.70 3.06 0.62 19.3 32.6 264 0.73 3.24 0.65 22.8 29.7 270 0.76 3.32 0.68 30.6 35.4 278 0.72 3.17 0.64 27.9 30.1 267 0.72 3.42 0.68 28.0 34.3 Mean S.D. N F.B.W 277 24.8 6 0.70 0.059 6 Final body weight 3.28 0.157 6 0.6.1 0.035 6 25.6 4.10 6 32.5 2.27 6 122 Appendix 9'- F 7 Study No. BMf Organ weight (Relative percentage of - individual values Sex Female t)ose level Sample D-1 30 Animals killed on schedule (Recovery) body weight) mg/kg Animal Number F.B.W (g) Bi*ain 4207 4208 4 209 4210 4211 4212 201 0.85 199 0.86 208 0.91 193 0.98 2-13 0.89 234 0.88 Mean S.D. N F.B.W 208 14.5 6 0.90 0.047 6 Final body weight Liver 5.93 6.55 6.20 6.03 5.82 5.49 6.00 0.358 6 Kidneys 0.72 0.81 0.75 0.76 0.74 0.79 0.76 0.033 6 Adrenals (XIO-3 24.9 27.0 22.0 27.5 25.2 28.9 25.9 2.43 6 Ovaries (XJO-3 30.7 29.8 35.0 38.1 33.4 30.8 33.0 3.18 6 123 Dose level Findings Animal number Lungs Dark reddish patch Liver -Scatterrtngof greyish dot/patch Ilypertrophy Greyish patch Dark reddish change Yellowish change kidneys Cyst Pyetectasis Greyish patch Blackish change Adrenals Dark reddish change Testes Atrophy No Iesion. P Present Appendix 10-H Individualmacroscopicfindings (29days) Set Kale Test article Sample D-L Study No.BKR143C 0 mg/kg (ol-) 0.1 mg/kg (It-) I mg/kg (21-) 10 mg/kg (31-) 30 aglkg 01 02 03 04 05 06 01 02 03 04 05 06 01 02 03 04 05 08 01 02 03 04 05 00 01 02 03 0 - -- - p --- - ---- -- --- - ---- -- --- -- -- - ---- -- --- --- -- - --- -p p p I - --- - --- --- --- -- -- -- --- - p p p I - - - - - - - - - - --- - - - - - - - - - - - - - - - - - - --- - - - - - - - - - - - - - - - - - - - - - - -- -- --- - ---- --- -- -- --- - --- p - - - - - - - - - - - --- - - - - - - - - - - - - - -- -- --p --- --- --- - -- --- -- - - -- -- --- --- -p - -- -- - ---- - --- -- - ---- --- -- -- --- - ---- - -- - --- - ---- -- --- --- -- - ---- - -- - - - - - - - --- --- - - - - - - - - - - - - - - - - Dose level Findings Animal number Lungs Dark reddish patch Liver -Scatterringof greyish dot/patch llypertrophy Greyish patch Dark reddish change Yellowish change kidneys cyst Pycleetasis Greyish patch Blackish change Adrenals Dark reddish change No lesion.P Present Appendix tO-F Individualmacroscopic findings (2&days) Sex Female Test article Sample D-1 Study No.BMR14 0 mg/k& (02-) 0.1 mg/kg (12-) 1 mg/kg (22-) 10 mg/kg (32-) 30 mg/ 01 02 03 *04 05 us 01 02 03 04 05 Do 01 02 03 04 05 .08 01 02 03 04 05 08 01 02 03 - --- -- --- -- -- --7 --- -- -- -- -- ---- -- --- - --- -- --- --- -- - -- --- --- -- -- --- -- --- -- -- -- p p p --- --- --- - --- -- --- -- -- -- -- --- --- -- -- --- -- -- --- --- p p - p - - - - - - - --- - - - - - - - - - - - - - - - - p - - - - - - - - - - - - - - --- - - - - - - --- - - - --- --- --- - --- -- -- --- -- --- - - - - - - - - - - - - 1-7 - - - - - - - - - - - - - - - - - - - - - - - - - - - -- - - - - - - - - - - - - - ---- -- -- -- --- -- ---- - --- - -- - Se% Dose level Findings Animal number Lung$ Dark reddish patch Liver Scatterringof greyish dot/patch flypertrophy Greyish patch Dark reddish change Yellowish change kidneys cyst Pyetectasts Greyish patch Blackish change Adrenals Dark reddish change Testes Atrophy No lesion.P Present Appendix 10-R-1 Individualmacroscopicfindings (Recovery) Test article : Sample U-1 Kate kg (ol-) 30 mg/kg -Tll--Y ol 08 es 10 11 12 07 08 10 it 12 01 Female 0 ag@kg--W2--) 08 09 10 11 12 07 Study No.BNR143C 30'silkg (42-) 09 09 le 11 12 - - - - - - - - - - -- -- -- -- -- -- -- --- --- -- 7- -- -- -- -- --- --- - --- -p -pp p -- -p p --p- ---- --- - - - - -- ----- ----- ---- --- -- -- ---- - --- - -- -- p -- --- -- -- ---- - ---- -- -- ---- - --- - - - --- -- --- -- - - - --- - --- -- p -- --- -- - p pp pp p--- -- - --- -- --- -- --- -- - - - - - - - - - - - --- Dose level(mg/kg) 30 Appendix 10-9-2 IndividualMacroscopicfindingsIn died sale rat (32days) Sex male Testarticle Sample D-1 Ailmal number Organ Macroscopic findings Thymus Dark reddish patch Stomach Atrophy Tarry contents Liver Ilemorrhage Scatterringof greyishdot./patch Study No Dose level Findings Animal number Liver sterogranuiona focal necrosis focal fatty change peripheral fatty change swellingof contrilobular hepatocytes eosinophillechange In centrilobularhepatocytes Ileart myocardial degeneration eyocardlal.flbrosts Spleen extromedullary hematopolesis Kidneys regenerated tubules dilated tubules infiltrationof lymphocytes dilatlonof pelvis chronic nephropathy Adrenals abnormality Brain abnormality Testes atrophy Appendix Lt-K Individualmicroscopicfindings (28days) Sex Kale Test article Sample D-1 Study No.BKR14 0 mg/kg (Ol-) 0.1 at/kg (li-) 1 mg/kg (21-) 10 ag/kg (31-) 30 mg/ 01 02 03 04 05 08 01 02 03 04 05 as 01 02 03 04 05 ol 01 02 03 04 05 06 01 02 03 + + t*t + + + + + --- --- -- -- --- -- -- --- --- - - -- - ---- --- -- --- -- --- -- 7 - -- - -- - ---- --- - -- ++ + - - - - - - - - - -- - - + 4+ + - ---- --- -- ---- --- - 4+ 4+ 4+ + + 4+ 4+ - -- - + - + + --- --- - + -- --- -- - - + - -- -+ -- - --- -- --- -- --- -- ----- Negative. :Very slight.+ Slight. 4+: Noderate. Appendix li-P Individualmicroscopic findlags (29days) Sex Female Testarticle Sample D-1 Study No.BNRI43C Dose level 0 mg/kg (02-) 0.1 mg/kg (12-). I mg/kg (22-) 10 eg/kg (32-) 30 mg/kg Findings Animal number 01 02 03 04 05 06 Of 02 03 04 05 as Of 02 03 04 05 06 01 02 03 04 05 08 01 02 03 0 Liver microtranutona focal necrosis + + + - --- -- --- --- -- -- --- -- --- -- - focal fattychange - --- - ---- --- -- -- --- - ---- - -- peripheralfattychange -- + + + + + + ++ + + swellingof contrilobular hepatocytes eosinophlitechange In - -- -- ---- --- -- -- --- 4+ if 4+ centrilobularhepatocytes Ileart myocardialdegeneration myocardialf.lbrosis - -- -- ---- --- -- -- --- - - -- -- - -- --- 4+ - -- Spleen extrasedullaryhematopolesis - - - - - - Kidney regen3erated tubules - + dilatedtubules -- -- -- inriitratioonf lymphocytes - - + dilationof pelvis -- -- -- chronicnephropalhy -- -- -- Adrenals abnormality -- -- -- Brain abnormality -- --- - - --- - -- - Ovaries abnormality -- -- -- - -- Negative. Very slight.+ Slight.4+: Moderate. Appendix 11-9-1 Individualmicroscopicfindings (Recovery) Test article Sample D-1 Study No.BKRi43 Sex Dose level Male 0 mg/kg (DI-) 39 mg/kg-Tit-7 FelAle 0 ng/kg (02-) 30 mg/kg (42-) Findings Animal number Liver microgranuiona focal necrosis focal fatty change -peripheral fatty change sw:llingof contrllobular epatocytes eosinophlitcchange In centrilobular hepatocytes fleart myocardial degeneration myocardial fibrosis Spleen extramedullary hematopolests Kidneys regenerated tubules dilated tubules Infiltrationof lymphocytes dilationof pelvis chronic nephropithy Adrenals abnormality Brain abnormality Testes atrophy Ovaries abnormality 07 us 09 to It 12 OT ol to [1 12 +-+ - - -- --- -- --- -- -- -- - --- -- -- - ++ ++ ++ -+ --- - - ++ 4+ 4+ . .. 4+ - -- -- - -- -- - -- -- -- - -- - - -- -- - -- -- - - - - - 4+ --- ---+ ---- ---- - --- -- --- -- 07 08 09 10 It 12 OT 08 09 10 11 t --- -- --- -- - + -- - + + +- - -- --- -- - - - - - - - - - - - * + + * 4+ -- -- -- . ... . - -- - -- -- - --- -- - -- -- - -- -- -- -- --- -- -- -- -- - - --- - - -- - -- - -- --- -- -- -- -- --- -- - -- -- -- -- - - - -- -- Negat Iva. :Very slight.+ :Slight. :Moderate. Appendix It-R-2 IndividualMicroscopicfiRdlOgsto died sale rat (32 days) Sal Kate Test.article Sample D-1 Dose level (mg/ig) 30 Animal number 4tQ9 Organ Liver Kidneys Brain Stomach Thymus lleart Spleen Adrenals Testes Microscopic findings Eos inophillechanie In centrilobut4rhePatocYtes + + swellingof centrilobularbepatocytes ++ Focal necrosis ++ Peripheral fatty change + Dilated tubules + Ilemorrhagicfoci + Ulcer and hemorrhage In glandular stomach + congestion + flemorrhage + No abnormality No abnormality No abnormality No abnormality + Slight,+ + Moderate Study No. Appendix 12-H Microscopicfindingsof the organs showed gross lesion (29 days) Sex Male Test article Sample D-1 Study N Dose level (mg/kg) Animal number Organ Macroscopic findings Microscopic findings 0 oto2 Kidneys cyst 0105 Lungs Dark reddish patch 0.1 1103 Kidneys PYCIeCt;kSIS Diluted tbbuies(+) llemorrhage(+) Dilationof pelvis(+) 1 2102 Kidneys Grayish patch Podal degenerationand calcification 1 regeneratedtubules(+) and lnflitra lymphocytes(+) 30 Alot Liver Scattering or grayish dot Focal necrosis(+) llypertropl)y Swelling or centrilobularbepatocytes( 4102 Liver Scatteringor grayish dot/patch No abnormalityexcept for oferogranulo peripheral rattychange (+).eosinoph centrtlobularhepatoCYLeS(++) 4103 Liver Ilypertrophy Scattering of grayisi)dot Sweiiingof centrilobularhepatocytes( I-ocalnecrosis(+) 4104 Liver 4105 Liver 410C Liver llypertropliy Scattering of grayish dot/patch flypertrophy Grayish patch IlYperLropl)y Scattering of grayish dot Sveitingof centrilobularhepatocytes( Focal necrosis(+) Swellingof centrllobularhepatocytes( Focal necrosis(+) Swellingor contrilobularhepatocytes( Fecai necrosis(++) Kidneys flypertrophy Dark reddish/Blackishdot Swellingof centrilobularliepatocytes( No abnormality Adrenals Dark reddish/Blackishdot No abnormality ;Very Slight :Slight. ++:Moderate Dose level (mg/kg) 10 Animal number 3208 Organ Liver so 4201 Liver 4202 Liver 4203 Liver 4204 Liver 4205 Liver ;t Very silght Kidneys 4206 Liver Silght, ++ Moderate Appendix 12-F Microscopic findingsof the organs showed gross leglon (28 days) . Sex 1'eivele Test article Sample 0-1 Study No Macroscopic findings Microscopic findings Dark reddish change Dark reddish change Ilypertrophy Yellowish change llypertrophy Dark reddish change llypertrophy Dark reddish change llypertrophy Dark reddish change Grayish patch flypertrophy Pyelectasts llypertrophy No abnormality except for microgranutoon Swelling of centrilobularhepatocytes( -nophille change In centrllobularheps No abnormality except for oferograoulosa nophille change In centrilobularhopat and focal necrosis(+) Swelling of ceotrllobularhepatocytes(+@ Peripheralfattychange (* ) Swelling of cestrilobularhepatocytes(++ No abnormality except for oferogranuiona eosinophillechange In centrilobularh nd..Peripheraflatty change(+) Sw:lllng of centrilobularbepatocytes(++ No.abnormality except for nicrogranuloss eostoophillechange to centrilobulr h Swelling of centrllobularhepatocyte:(+) No abnormalityexcept for eosinophillec in contrilobularhepittocytes(++) No abnormalityexcept for eosinophlitcc In centrilobularhepatocytes(++) Swelling of centrilobularhep4tocytes(++ Dilationof pelvis(+) Swelling of cestrilobularhapatocytes(++ Dose level (mg/kg) Male 0 30 Animal number -.0111 Organ Testes Liver Female 30 4109 4109 4110 4111 4112 4207 4208 Liver Thymus Stomach Liver Liver Liver Liver Liver Liver :t: YerY sIlght : Dead anleal 4209 Liver 4210 Livei 4211 Liver 4212 Liver Silght, ++ : Moderate Appendix 12-R Microscopic findingsof the organs showed gross lesion (Recovery) Test article : Sample D-1 Macroscopic findings Microscopic findings Study N Atrophy Dark reddish change Grayish patch llypertrophy Dark reddish patch Atrophy Tarry contents/liesorrhage Scatteringof grayishdot/patch Ilypertrophy Dark reddish change Grayish patch .'Atrophy(++) No abnormalityexcept for swellingof c hepatocytes(+-+).anodosinophille chan centrilobularhopatocytes(+)and afer Focal necrosis(+) sweiiingor centrllobularbepatocytes(+ Congestion(+)and hemorrhage(+) No abnormality Ulcer and hemorrhage In glandular stoma Focal necrosis(++) Svellingof centrilobularbepatocytes(+ No abnormality except for eferograouloo swelling of contrilobularhepatocytes eminophille change In centrilobular Focal necrosis(+) Dark reddish change Dark reddish change flypertrophy Dark reddish change Dark reddish change Dark reddish change Grayish patch No abnormality except for oferogranuloo of contrilobularbepatoertes(++)and change In contrilobularhepatocytes(+ No abnormality except for microgranuion focal necrosis(+)and oosloophilicch -rilobularhepatocytes(+) Swelling of centrilobularhepatocytes(+ No abnormality except for oferograouloo of centrilobularhepatocytes(+) and c4 change In centrilobularbepatocytes(+ No abnormality except ror aterogranuloo of centrilobularhepatocytes(++)and change In centrilobularhepatocytes(+ No abnormality except ror microgranuion or centrilobularhepatocytes(++)and change In centrilobularhepatocytes(+ Focal necrosis(+) - '0@@. PHOTOGRAPHS (HisoPathologicaPlictures) let 4i6 P4- e 4e 0- Is lw IKil IV" t 0 i. Photo-1 Liver of a male rat of control. (Anim 1 centrilobular region. Showing normal No. 0103) - H.E.stain X 180 gaqr; 40 Photo.2 male rat given 30 mg/kg. Liver of a (Animal No. 4105) of cent rijobular hepatocytes are Eosinophilic change and swelling noted. H.E.stain X 1BO Photo.3 Liver of a male rat given 30 mg/kg. (Animal No. 4105) Focal necrosis Is noted. H.E.stain X 180 LTiw Photo.4 Liver of a male rat given 30 mg/kg. (Animal No. 4106) Peripheral fatty change is found. H.E.stain X 180 2