Document QgQw2OVa6V40dNvmXdEnR1yQ5

Attachments to Letter to C. Auer dated May 4, 2000 Perfluoroctane Sulfonate Studies Teratology Pilot Teratology Study in Rats, T-3551 Final Report, May 13, 1983, 3M Reference T3551.12 004207 HAZLETON LABORATORIES AMERICA, INC. 9 2 0 0 L E E S B U R G T U R N P IK E . V IEN N A . V IR G IN IA 221BO, U.S.A. May 13, 1983 William C. McCormick, III Toxicologist Toxicology Services Medical Department/3M 220-2E 3M Center St. Paul, Minnesota 55101 Dear Bill: Enclosed are four copies of the final report for Project No. 154-158, Pilot Rat Teratology Study with T-3351. With copy of this letter, I am forwarding one copy of the report to Marshall Johnson. If you have any comments or questions, please contact me. Best regards. Sincerely, LTW:sm Enclosure cc: E. Marshall Johnson LAWRENCE T. WETZEL, Ph.D. Staff Scientist Department of Toxicology PHONE C703] 893-5400. TELEX 89 9 4 3 6 [HAZLABS VINA}, CABLE HAZLABS WASH OC C04208 @ H AZLETO N ,I LABORATORIES AMERICA. INC. aaoo l c e s e u r s t u r n p ik e . Vie n n a , v ir g in ia aaiso. u s a . -- ---- PILOT TERATOLOGY STUDY IN RATS T-3551 FINAL REPORT Submitted to 3M Company S t. Paul. Minnesota May 13, 1983 004209 @ H A ZLETO N LA BO RA TO RIES AM ERICA, INC. 9 8 0 0 I f C M u R Q T U AP N P I K I . vVilSSNNINs A . VIM Q IN IA 8 8 1 0 0 . U.8u. SUBJECT: Pilot Teratology Study in Rats Project No. 154-158 We, the undersigned, hereby declare that the work was performed under our supervision, according to the procedures herein described. Study Director: i t i u M tt________ O W R E H E E T T WETZEL, Ph.D. Staff Scientist Department of Toxicology Laboratory Supervision: Teratology: a tU F \ a t v n . uursuvsuiv, r i i . w Rodent Toxicology I Department of Toxicology RUTH S. Du RL'OO, B.S. Research Associate Department of Toxicology Report Preparation: S%LUACL A/. W w tu ,iJ x L LAURA H. MISTRETTA, B.S. Group Leader Scientific Resources Department tlh 004210 (glH A ZLETO N LABO RATO RIES AM ERICA, INC. a aa no no aLiEtEMs aiu>naonvtaulpanupaiikxae . nV iaenn in a , v i p o i n i a a a ia a . u j a TABLE OF CONTENTS SUMMARY INTRODUCTION CONTROL AND TEST MATERIALS TEST ANIMALS METHODS Mating Period Groups and Dosage Levels Compound Preparation and Administration Maternal Observations and Records Cesarean Sacrifice and Tissue Preservation Statistical Analyses Specimen, Raw Data, and Final Report Storage RESULTS - MATERNAL DATA Clinical Signs Mortality Body Weights Food Consumption Gross Pathology Uterine Weights RESULTS - CESAREAN DATA Pregnancy Rates, Corpora Lutea, Uterine Implantations, and Implantation Efficiency Fetal Viability, Weight, and Sex Fetal Development TABLES Table 1 - Clinical Observations During Gestation Table 2 - Mean Maternal Body Weights and Body Weight Change Table 3 - Mean Food Consumption Values Table 4 - Summary of Gross Pathology Findings Table 5 - Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Key to Table 6 Table 6 - Summary of Ovarian, Uterine, and Litter Data Table 7 - Mean Incidence Values for Visceral and Skeletal Findings Key to Table 8 Table 8 - Mean Incidence of Visceral and Skeletal Findings per Litter 1 3 3 4 4 4 5 5 6 7 8 9 10 10 10 10 11 11 11 12 12 12 12 14 15 16 17 18 19 20 21 23 24 004211 @ HAZLSTOfM LABO RATO RIES AM ERICA, INC. aa aanono L tiks aa aa iuina on tt iuaainl.iaAiok 'b . vv/iibs inu kn a , v i r g i n i a a a is o . u .S ja. TABLE OF CONTENTS - CONTINUED APPENDICES Key to Appendix 1 Appendix 1 -Individual and Mean Maternal Body Weights Appendix 2 -Individual and Mean Food Consumption Values Appendix 3 -Individual Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Appendix 4 -Individual andMean Ovarian, Uterine, and Litter Data Appendix 5 -Individual and Mean Live Fetal Data Appendix 6 -Individual Visceral Findings for Each Litter Appendix 7 -Individual Skeletal Findings for Each Litter Appendix 8 -References Page 25 26 28 31 34 36 38 39 42 004212 HAZLETON LABORATORIES AMERICA, INC. 9 2 0 0 L E E S B U R G T U R N PIK E. V IEN N A. V IR G IN IA 221SO. U.S.A. SPONSOR: 3M Company DATE: May 13, 1983 MATERIAL: SUBJECT: FINAL REPORT Pilot Teratology Study in Rats Project No. 154-158 SUMMARY T-3351, suspended in corn oil at concentrations intended to deliver 1, 5, 10, or 20 mg/kg/day (Groups 2, 3, 4, and 5, respectively), was administered by oral intubation to four groups of mated female Sprague-Dawley SD rats on gestation Days 6 through 15. A fifth group (Group 1) of mated females served as the control group and received the vehicle only. No Group 5 females survived to cesarean section on gestation Day 20. Treatment-related clinical observations in Groups 4 and 5 included hunched and thin appearance of the animals. In addition, tremors and convulsions as well as prostration and anorexia were noted in Group 5 animals. Body weight gain of the dams during gestation was less in Groups 3, 4, and 5 compared to control, with a significant reduction in weight gain occurring in Groups 4 and 5. A significant decrease in food consumption occurred in Groups 4 and 5. Apparent treatment-related ulcerated areas on the glandular and/or nonglandular stomach mucosa were found during necropsy of Group 5 females. 004213 ) w u i n LA BO RA TO R IES AM ERICA, INC. a a o o iLiiKKMi suu nR a t u p n pp ik bs . v iii n nn a . v i p q i n i a a a io a . u .s .a . 2- - 154-158 Analysis of individual and mean uterine data demonstrated no effect of T-3351 on fetal mortality or fetal viability. Mean fetal weight was comparable for either sex in all groups. External, visceral, and skeletal abnormalities were generally similar for all groups. One Group 3 fetus was found to have a small eye upon external examination. Another Group 3 fetus had fused hind limbs and anal atresia. Skeletal examination confirmed the external limb anomaly. 004214 b W I W LABORATORIES AMERICA. INC. a a o o L s c s s u n o t u iwp<k *, v i c m n a . v i r o i n i a a a i a a . m i * -3- 154-158 INTRODUCTION This study was designed to determine the maternal and embryo/ fetal toxicity of T-3551 when administered by gavage to pregnant rats during the period of fetal organogenesis for the purpose of setting dosage levels for an expanded teratology study. The rats were placed in breeding on January 11, 1983, and cesarean sections were completed on February 3, 1983. This report presents the methods and results from this study. CONTROL AND TEST MATERIALS The vehicle and control material, Duke's Corn Oil (C. F. Sauer Co., Richmond, Virginia), Lot 80235, was received on May 28, 1982 and was stored at room temperature. The test material, T-3351, a white powder, was received from the sponsor on January 5, 1983, and was stored at room temperature. The test material was assumed to be 100% active compound. Information on methods of synthesis and stability, as well as data on composition or other characteristics which define the test material, are on file with the sponsor. 004215 )n U A i U C U U J I LA B O R A T O R IES A M ERIC A, INC. a a o o L S f ia a u f lG t u r n p ik k . v ic n m a . v ia q in ia a a i o . u .a .A . -4- 154-158 TEST ANIMALS Sexually mature, cesarean-derived Sprague-Dawley Crl:CD(SD)BR rats were chosen for this study because they are sensitive to a number of agents which are known to be embryotoxic and/or teratogenic. Rats have historically been used in safety evaluation studies of this type and are required by the regulatory agencies. Thirty-six male and thirty-six female thirteen-week old rats were received from Charles River Breeding Laboratories, Kingston, New York, on December 22, 1982. The rats were held in quarantine for two weeks, during which time a health status examination was performed by a staff veterinarian. The rats were housed one male and one female per cage during breeding. Following confirmation of mating, the females were housed individually in elevated wire-mesh cages with food (Purina Rodent Laboratory Chow 5001) and tap water available ad libitum. The females were uniquely identified by ear tag after mating was confirmed. Tempera tures in the study room ranged from 71 to 75F with a relative humidity between 37 and 65%. METHODS Mating Period During the mating period, one female was paired with one male until mating was confirmed or until two weeks had elapsed. Daily vaginal examinations of each female were performed to detect the presence and 004.216 ')n / - \ ^ U C L L J I X LA B O R A T O R IES A M ERICA , INC. a O O L E E S B U R G T U R N P IK E . V IE N N A . V IR G IN IA S 8 1 B O . U .S A . -5- 154-158 viability of sperm or the presence of a copulatory plug. The day of observation of sperm or copulatory plug was designated as "Day 0" of gestation. Groups and Dosage Levels Upon confirmation of mating, each female was assigned to one of the following groups. Group 1 2 3 4 5 Dose3 mg/kg/day 0 1 5 10 20 Number of Females 6 6 7 7 7 3 Based on individual animal body weights at each weighing interval during the dosing period. The females were placed in the dose groups one at a time beginning with the high-dose group and continuing sequentially through the control group until all mated females were assigned to a group. All males were sacrificed via carbon dioxide asphyxiation and discarded without necropsy once a sufficient number of matings were confirmed. Compound Preparation and Administration Suspensions of the test material in the corn oil vehicle were prepared on a weight per volume basis. The required amount of compound CQ4217 LA B O R A T O R IES A M ERIC A. INC. S 8 0 0 iL (B sE 9uS Ui RtoCr3tT iUiRBKN iPeIiKva* . VwiIsSiuNiNkA . V I R G I N I A 8 8 1 8 0 . U .8 .A . 6- - 154-158 for each level was weighed on an electronic Arbor 126 balance and trans ferred to a 7 ml homogenizer. Approximately 5 ml of corn oil was added and the test material was ground into a fine suspension with a teflon pestle. This suspension was transferred to a beaker and any remaining compound in the homogenizer was rinsed into the beaker. Additional corn oil was added to the suspension to produce the desired concentration and the suspension was mixed on a stirrer for approximately ten minutes. Prepared suspensions were mixed with a magnetic stirrer during dosing. Females were given the appropriate dosing suspension or vehicle by oral intubation on a daily basis beginning on Day 6 and continuing through Day 15 of gestation. The test material was administered orally because of the relative ease and accuracy of dosing. Fresh dosing suspensions were prepared weekly and stored under refrigeration (approximately 41F). Samples of each test mixture as well as the vehicle were sent to the sponsor for analysis. Maternal Observations and Records All animals were examined twice daily for mortality and mori- bundity (from day of receipt through study termination) and once daily for clinical signs of toxicity and pharmacologic effects (throughout gestation). Body weights and food consumption were recorded on Days 0, 6, 8, 12, 16, and 20 of gestation. 004218 LA B O R A T O R IES AM ERICA, INC. a9 2a0n0n>LMe eas auu RioOntriuironmpbiikvei. wV iiennjn>a , v i r g i n i a 2 2 1 9 0 . u.a.A . -7- 154-158 Cesarean Sacrifice and Tissue Preservation A gross necropsy was performed on all animals (found deads, moribund sacrifices, and terminal sacrifices). The uterus and ovaries from found dead animals and animals sacrificed because of moribundity were removed and examined for the number of implantation sites and corpora lutea, respectively. Prior to gestation Day 20, surviving females were assigned random numbers and all personnel performing cesarean sections and/or external, visceral, or skeletal examinations of the fetuses were unaware of the dose level from which the animals were derived. On Day 20 of gestation, all surviving females were weighed and sacrificed by carbon dioxide asphyxiation. The uterus from each female was weighed and examined for the number and placement of uterine implantation sites, number of live and dead fetuses, early and late resorbing fetuses, and any abnormalities. The uterus of each animal was reweighed after the contents were removed. The ovaries were examined for the number of corpora lutea. Each live fetus was sexed, weighed, and examined for external abnormalities. Findings were recorded. Beginning at the ovarian end of the right uterine horn, the first six fetuses, regardless of sex, were selected for further evaluation. After the external examination was completed, the first fetus was identified with a tag and fixed in Bouin's solution for soft tissue evaluation (Wilson, et ^ . , 1965). The next two fetuses were eviscerated, tagged, and processed for skeletal examination using a 004219 H A ZLETO N LABORATORIES AMERICA, INC. 9 8 0 0 L E E S B U R G T U R N P IK E . V IE N N A . V IR G IN IA 8 8 1 8 0 . U .8 .A . 8- - 154-158 technique modified from that reported by Staples (Staples and Schnell, 1964). This sequence was repeated until a maximum of six fetuses had been selected. Statistical Analyses Mean maternal body weight gains (Days 6-16 and 0-20), total food consumption, gravid uterine weight, terminal body weight minus gravid uterine weight, resorption incidence, percent males, and fetal viability were analyzed in the following order. Levene's test for homogeneity of variances (Levene, 1960; Draper and Hunter, 1969) was performed and if the variances proved to be homogeneous, the data were analyzed by one-way classification analysis of variance (ANOVA) (Winer, 1971). If the variances proved to be heterogeneous, a series of transformations was performed until variance homogeneity was achieved. These transformations were: log1Q, square (X ), square root (X2), reciprocal (1/X), angular (arcsine X*5), and rank, in that order. If rank transformation was ineffective in removing variance heterogeneity, ANOVA of ranked data was completed. If ANOVA of untransformed or transformed data was signifi cant, Dunnett's t-test (Dunnett, 1955 and 1964) was used for control vs. compound-treated group mean comparisons. If ANOVA was not significant, the analysis was complete. Mean number of fetuses with visceral variants per litter and skeletal variants per litter were analyzed by Fisher's exact test for proportions (Fisher, 1950). 004220 LA B O R A T O R IES A M ERIC A, INC. a9 3an0 n0 L>bE 3iSh9 iU iRoGnTriUiOP NmPBIiKi/Ec. VuiIbEiNuiNlA . V l A Q I N I A 3 3 1 9 0 . U .S .A . -9- 154-158 In addition to the above data, analysis of covariance (ANCOVA) (Winer, 1971) was used to analyze mean fetal weights. The litter was used as the experimental unit. Levene's tests and ANOVA were evaluated at the 5.0% one-tailed probability level. Control vs. compound-treated group mean comparisons were evaluated at the 5.0% two-tailed probability level. Statistical references are appended to this report, and statisti cally significant differences, as indicated by the aforementioned tests, are designated throughout this report by the term "significant" and/or as follows: S+ * Significantly higher than the control value. S- = Significantly lower than the control value. Specimen, Raw Data, and Final Report Storage All specimens, raw data, and the final report are stored in the archives of Hazleton Laboratories America, Inc. 004221 LA B O R A T O R IES A M ERIC A , INC. a so on on LL S B MS Bu fu ln So tt iujrsnm pb ikk ei , vvikI nN Nn a , v i a o i n i a a a i a o . u .b a . - 10 - 154-158 RESULTS - MATERNAL DATA Clinical Signs Summary clinical signs are presented in Table 1. Treatment-related clinical observations were noted in the Groups 4 and 5 animals during the treatment and posttreatment phases. These included hunched and thin appearance and rough haircoat in both groups, and tremors, convulsions, prostration, and anorexia in the Group 5 females. Mortality All Group 5 females were found dead or sacrificed in a moribund condition prior to cesarean section on Day 20 of gestation. All other animals survived to Day 20 of gestation. Body Weights Individual and mean body weights and body weight changes are presented in Appendix 1. Mean weight values and body weight changes are presented in Table 2. Lower mean body weight values were noted in the treated groups. Decreased body weight gain during gestation was noted in Groups 3, 4, and 5, with significant decreases noted in Groups 4 and 5. (Due to the number of deaths in Group 5, body weight change values could not be determined for Days 0-20.) 004222 'n u A u c u -- ii\ LA BO RA TO R IES AM ERICA, INC. sA oB tDvOi LE E S a U iReOntTiUiRaiNuPaIiKi/EB. wV iIsEiNukN A . V I R O I N I A 8 8 1 * 0 . U .S .A . 11- - 154-158 Food Consumption Individual and mean food consumption values are presented in Appendix 2. Mean food consumption values are presented in Table 3. Decreased food consumption was noted in all treated groups in a dose-related fashion. Significantly lower total food consumption values (Days 0-20) were noted in Groups 3 and 4. (Due to the number of deaths in Group 5, total food consumption values could not be determined.) Gross Pathology Summary gross pathology findings are presented in Table 4. Treatment-related findings were noted in the Group 5 rats and included ulcerated-like areas on the glandular or nonglandular stomach mucosa. Uterine Weights Individual and mean uterine weights are presented in Appendix 3. Summary uterine weights are presented in Table 5. The terminal body weights minus the gravid uterine weights were lower in the treated groups when compared to the control group; however, no statistical differences were noted. The gravid uterine weights were comparable between the control and treated groups. 004.223 LA BO R A T O R IES A M ERIC A, INC. a a o o L ce sa u R O t u r n r i k *. v i b n n a , v i r g in ia aaiao. u ..a . 12- - 154-158 I RESULTS - CESAREAN DATA Individual and mean ovarian, uterine, and litter data are presented in Appendix 4. Individual and mean live fetal data are presented in Appendix 5. Summary ovarian, uterine, and litter data are presented in Table 6. Pregnancy Rates, Corpora Ltea, Uterine Implantations, and Implantation Efficiency Pregnancy rates were 100 percent for Groups 2, 3, and 5; 83 percent for Group 1, and 86 percent for Group 4. The mean number of corpora ltea and uterine implantations were comparable for all groups. The implantation efficiency for the control group was low; however, this may be attributed to two animals which had implantation efficiencies of 38.4 and 35.7 percent. Fetal Viability, Weight, and Sex The mean incidence of resorptions was similar for Groups 1-4. The mean incidence of fetal viability was comparable between the control group and Groups 2, 3, and 4, as were mean fetal weights and the percent males per litter. Fetal Development Individual visceral findings are presented in Appendix 6; indivi dual skeletal findings are presented in Appendix 7. Mean incidence values 004224 LA B O R A T O R IES A M ERIC A, INC. e o a L K K ia u n a t u h n p i k e . v i i n n a v i r g i n i a a a i a . u.a^t. - 13 - 154-158 for visceral and skeletal findings are presented in Table 7 (based on number of fetuses) and Table 8 (based on number of litters). External anomalies were noted in two Group 3 fetuses at cesarean section. These included a small right eye in one fetus, and short and thickened tail, no external genital/anal opening and hind legs which were joined from the pelvis to the feet in the other fetus. All remaining fetuses appeared normal. There was no apparent treatment-related increase in the incidence of visceral or skeletal variants or anomalies. Incompletely descended testes were noted in two Group 4 fetuses. Skeletal anomalies noted in one Group 3 fetus included a small pubis, fused ischia, and cojoined hind limbs. This fetus was the one for which the tail and hind limb external abnormalities were noted. Observation Number which appeared normal Alopecia Hunched Rough halrcoat Bloody crust Thin Urine stains Tremors Convulsions Prostrate Anorexia Soft feces Bloody discharge from vagina PCillointicTaelraOtoblosegryvTaStaitboulndeys 1DofurTin-3g55G1esItnatiRoants 0-5 G6B-r1oa5uyps 1 TBTZff 65 5 11 0-5 G6D-r1oa5uyps 2 16-20 65 6 0-5 G6b-r1oa5yups 3 1630 77 7 1 0-5 G6b-r1oa5yups 4 16-2 72 3 11 21 11 21 11 154-158 G6D-r1oa5uyps 5 630 6 11 5 6 6 7 4 1 3 3 6 1 5 4 6 5 4 3 3 5 1 -- 042 Ci Table 2 Mean Maternal Body Weights and Body Weight Change Pilot Teratology Study of T-3551 1n Rats 154-158 Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. n_____________________ Body Weight (g)_____________________ Day^of Gestation 0 -----5----- s-- -- -- ----IB----- 20 231.67 256.38 260.96 280.92 303.22 355.36 13.831 9.187 12.574 13.102 20.915 33.795 216.53 243.78 246.60 266.08 289.38 348.43 8.640 12.094 11.064 18.994 16.705 23.213 231.16 253.44 253.79 262.24 291.21 350.90 10.085 10.692 8.578 9.340 9.111 16.306 228.75 254.33 254.85 262.15 266.08 333.75 19.401 18.145 20.020 21.046 24.184 26.277 228.29 245.06 238.70 202.73 175.05 14.039 13.923 9.242 6.388 5.720 - Body Weight Change (g) pays of Gestation "T P 6 0-20 46.84 16.308 123.60 26.087 45.60 9.297 131.90 15.259 37.77 9.800 119.74 15.426 4.37S' 22.593 105.00 13.301 -64.50S" 17.593 - | 1t--n* 0042 vF Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. Table 3 Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 23.73 2.501 20.88 4.330 21.47 1.923 21.61 2.006 19.31 2.741 Food Consumption (qm/day) Day of Gestation 8 12 lb 20.13 4.132 21.55 3.597 21.10 4.145 18.50 4.159 19.85 2.874 19.27 2.430 16.47 2.735 16.60 1.880 17.66 1.269 16.90 1.615 17.46 5.336 10.91 3.879 14.13 2.743 4.74 4.028 0.32 0.240 20 26.05 4.284 27.00 2.211 24.81 1.585 22.11 3.021 - 154-158 Total Food Consumption Days of Gestation 0-20 457.12 58.499 410.80 29.452 397.83$23.480 365.09$41.840 - Table 4 Summary Gross Pathology Findings Pilot Teratology Study of T-3551 in Rats 154-158 ORGAN AND DESCRIPTION 7Group: 1 2 3 4 Dose Level (mg/kg): T T ~T> Tff 5 Number of animals examined Number of animals with no gross lesions 66 7 77 65 772 STOMACH Ulcerated-like areas/dark foci onglandular mucosa Smooth glandular mucosa Ulcerated-like areas onnonglandular mucosa 3 1 1 VAGINA Red discharge 1 HEART Enlarged atria 1 LIVER Mass on median lobe adhered to diaphragm 1 DIAPHRAGM Small hole 1 0042 10 <4 004230 Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Table 5 Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats 154-158 Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. Terminal Body Weight 355.36 33.795 348.43 23.213 350.90 16.306 333.75 26.277 - Gravid Uterine Weight 62.82 28.982 72.45 8.097 74.71 12.846 74.78 12.312 - Terminal Body Weight Minus Gravid Uterine Weight 292.54 9.634 281.93 30.878 276.19 10.211 258.97 19.333 - - 19 Key to Table 6 154-158 Pregnancy Rate (percent) (number of pregnant females/number of females which showed evidence of copulation) x 100. Survival Rate (percent) = (number of females surviving to Day 20/number of females which snowed evidence of copulation) x 100. Mean Implantation Efficiency (percent) = Group mean of ([resorptions per litter/corpora lutea per litter] x 100). Mean Incidence of Resorptions (percent) = Group mean of ([resorptions per litter/implantations per litter] x 100). Mean Inci dence of Fetal Vi abi 1ity (percent ) Group mean of ( [live fetuses per litter/implantations per litter] x 100). 004231 C04232 Table 6 Summary of Ovarian, Uterine, and Utter Data Pilot Teratology Study of T-3551 in Rats Group: 1 Dose Level (mg/kg): -- 0 Number of females with mating confirmed (%)Number of females pregnant Pregnancy rate Number of females surviving to Day 20 Survival rate 6 5 83 6 100 Mean number of: Corpora lutea Implantations Resorptions Fetuses - dead - live Indices calculated on a per litter basis: (%)Mean implantation efficiency (%)b Mean resorption incidence Mean incidence of fetal mortality (%) (%)Mean incidence of fetal viability Live fetuses Mean body weight (g) Males - unadjusted - covariance adjusted Females - unadjusted - covariance adjusted Percent males 15.2 11.0 0.6 0.0 10.4 70.0 4.1 0.0 95.9 3.70 3.65 3.48 3.48 42.3 2 -- r~ 6 6 100 6 100 15.5 13.7 0.5 0.0 13.2 88.2 3.6 0.0 96.4 3.35 3.36 3.25 3.26 50.7 a The ovaries of two animals were too autolyzed to count corpora lutea. k Data analyzed following rank transformation. 3 ------- 5 " 7 7 100 7 100 15.7 13.7 1.0 0.0 13.0 89.1 7.0 0.0 93.0 3.51 3.53 3.34 3.34 54.6 154-158 4 " 10" 7 6 86 7 100 16.3 13.5 0.3 0.0 13.2 83.1 2.6 0.0 97.4 3.43 3.47 3.30 3.29 59.2 5 2 7 7 100 0 0 14.1 10. - - - 004233 Table 7 Mean Incidence Values for Visceral and Skeletal Findings Pilot Teratology Study of T-3551 in Rats Group: Dose Level (mg/kg): 1 0 Visceral Examinationi Number of fetuses examined Number and percent of fetuses having one or more visceral variants 10 1/10.0 Number and percent of fetuses with: Dilated ureter(s) Undulated ureter Incompletely descended testis 1/10.0 0/ 0.0 0/ 0.0 Number and percent of fetuses having one or more visceral anomalies 0/ 0.0 Skeletal Examination Number of fetuses examined Number and percent of fetuses having one or more skeletal variants 18 13/72.2 Number and percent of fetuses with: Skull - incomplete closure (less than 75%) - interparietal incomplete ossification - supraoccipital - incomplete ossification - nonfused - nonossified - hyoid - incomplete - nonossified - pari tais irregularly ossified - zygoma incomplete 0/ 0.0 6/33.3 7/38.9 0/ 0.0 1/ 5.6 0/ 0.0 0/ 0.0 3/16.7 0/ 0.0 2 --- 1 12 2/16.7 2/16.7 0/ 0.0 0/ 0.0 0/ 0.0 24 15/62.5 3/12.5 13/54.2 13/54.2 2/ 8.3 0/ 0.0 0/ 0.0 8/33.3 9/37.5 2/ 8.3 3 --- 5 14 2/14.3 1/ 7.1 1/ 7.1 0/ 0.0 0/ 0.0 28 22/78.6 0/ 0.0 13/46.4 14/50.0 2/ 7.1 0/ 0.0 4/14.3 0/ 0.0 0/ 0.0 0/ 0.0 154-158 4 T 12 2/16.7 0/ 0.0 0/ 0.0 2/16.7 0/ 0.0 24 14/58.3 2/ 8.3 6/25.0 7/29.2 0/ 0.0 1/ 4.2 1/ 4.2 1/ 4.2 2/ 8.3 0/ 0.0 Table 7 - Continued Mean Incidence Values for Visceral and Skeletal Findings Pilot Teratology Study of T -3551 in Rats Group: Dose Level (mg/kg): 1 0 Rib cage - sternebrae bipartite - ribs irregularly ossified Vertebral column - thoracic centra - incomplete - nonfused - lumbar arch irregularly ossified - sacral arch - incomplete - nonossified - caudals 3 Pelvic girdle - ischium incomplete - ischium and pubis irregularly ossified - pubis nonossified 1/ 5.6 0/ 0.0 1/ 5.6 4/22.2 0/ 0.0 0/ 0.0 1/ 5.6 1/ 5.6 1/ 5.6 0/ 0.0 1/ 5.6 Bone alignment Sternebrae checkered Fused ribs, malaligned thoracic centra and arches Angulated rubs Extra rib(s) Femur short 1/ 5.6 0/ 0.0 1/ 5.6 2/11.1 0/ 0.0 Number and percent of fetuses having one or more skeletal anomalies 0/ 0.0 Number and percent of fetuses with: Bone alignment Fused ischia and cojoined hind limbs; pubis small 0/ 0.0 2 --- 1 0/ 0.0 0/ 0.0 0/ 0.0 0/0.0 0/ 0.0 1/ 4.2 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 3/12.5 1/ 4.2 0/ 0.0 0/ 0.0 0/ 0.0 3 5 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 2/ 7.1 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 2/ 7.1 0/ 0.0 0/ 0.0 1/ 3.6 17/60.7 1/ 3.6 1/ 3.6 1/ 3.6 1/ 3.6 154-158 4 10 3/12.5 1/ 4.2 2/ 8.3 0/ 0.0 0/ 0.0 1/ 4.2 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0.0 1/ 4.2 0/ 0.0 0/ 0.0 3/12.5 3/12.5 0/ 0.0 0/ 0.0 - 23 Key to Table 8 154-158 Mean Incidence of Visceral Anomalies (percent) * Group mean of ([number of fetuses with anomalies per litter/number of fetuses examined visceral 1y per litter] x 100). Mean Incidence of Viscera1 Variants (percent) * Group mean of ([number of fetuses with variants per litter/number of fetuses examined viscerally per litter] x 100). Mean Incidence of Skeletal Anomalies (percent) = Group mean of ([number of fetuses with anomalies per litter/number of fetuses examined skeletally per litter] x 100). Mean Incidence of Skeletal Variants (percent) = Group mean of ([number of fetuses with variants per litter/number of fetuses examined skeletally per litter] x 100). 004235 Table 8 Mean Incidence of Visceral and Skeletal Findings per Litter Pilot Teratology Study of T-3551 in Rats Group: 1 Dose Level (mg/kg): " Visceral Number of litters examined Mean values calculated on a per litter basis Number of fetuses with variants Incidence of variants Niunber of fetuses with anomalies Incidence of anomalies 5 0.2 10.0 0.0 0.0 Skeletal Number of litters examined Mean values calculated on a per litter basis Number of fetuses with variants Incidence of variants Number of fetuses with anomalies Incidence of anomalies 5 2.6 71.6 0.0 0.0 ~ T2~ 6 0.3 17.5 0.0 0.0 6 2.5 62.5 0.0 0.0 T3 7 0.3 14.6 0.0 0.0 7 3.1 78.6 0.1 3.6 4 ~nr 6 0.3 16.7 0.0 0.0 6 2.5 58.3 0.0 0.0 - 25 - Key to Appendix 1 P = Pregnant NP = Not Pregnant C * Cesarean Delivery, Preceded by Day of Gestation FD = Found Dead, Preceded by Day of Gestation MS * Moribund Sacrifice, Preceded by Day of Gestation 154-158 004237 MNautmerbnearl 222222000000777777433433176089 MS.eDan. 222222000000777777444444745368 MS.eDan 22222220000000777777755555531532460 MS.eDan. NDaattlenqof 111111//////111111112312//////888888333333 111111//////111111223111//////888888333333 1111111///////11111111123412///////88888883333333 Disposition NPPPPPP,,,,,. 222222000000------CCCCCC PPPPPP,,,,,, 222222000000------CCCCCC PPPPPPP,,,,,., 22222220000000-------CCCCCCC InPdiliovitdTuaelraatonldogMyAeapnSpteuMnddayitxeor1nfaTl-3B5o5d1y HIneiRghattssa -- o--------- Bo6dy Height (gra8ms) - Day ofIFGestation 16 Group 1 - 0 mg/kg (222222053212127486......233409) (222222345556036948......153650) (222222356467004657......839088) (222222368869478908......545089) (222333388021960098......196789) 23131..87361 2596..13887 21620..59764 28130..19022 32030..29215 Group 2 - 1 mg/kg 222222031211600226......148955 222222433546002550......444870 222222464633700009......463797 222222485965152269......111642 223223780197015107......113396 2186..65430 21423..07984 24161..60604 21686..90984 21869..73085 Group 3 - 5 mg/kg 222222224221333586688.......3485678 222222244755540804289.......1326577 222222254565643617250.......3379300 222222266546767929105.......1424286 222222399888801528469.......1763693 21301..01865 21503..64942 2583..57798 2692..32440 2991..12111 ' 20 (233333488621858281......384088) 33535..73965 333333363283469839......200860 32438..24133 333333344453482493555.......5365347 31560..39006 154-158 H6e-i1g6httI Change 0-20 46264(911443......128333) 111(1492530732965......986160) 4166..83408 12263..06807 344355950448......379665 111111521224806861......112569 495..26907 11351..29509 42434235996353.......0849968 11111194021229975042.......1157701 397..87070 11159..47246 * Only pregnant animals Mere Included In group mean Numbers within parentheses were excluded from mean calculations. b Data analyzed follow ing rank transform ation. C0423 05 lNautemrbnear1 MDaatteingof 22222220000000777777765656651728039 1111111///////11111111123312///////88888883333333 MS.eDan. 22222220000000777777766666675467908 1111111///////11111111111223///////88888883333333 MS.eDan. Disposition NPPPPPPP,,,,.,, 22222220000000-------CCCCCCC PPPPPPP,,,.,,, 21111117049788-------MFFFFFFDDDDDDS IPndiliovitduTaelrAaatpnopdleongMdyiexanS1tuMd-aytCeoronfnatTiln-u3Be5od5d1y WIneiRgahttss9 0 (222222220314159674643.......4624190) 22198..47061 222222212222531416463.......1242597 22184..20399 Bo6dy Weight (gra8ms) - Day of1G2 estation 16 Group 4 - 10 mg/kg (222222232446679826862.......2359998) (222222235376268727939.......1239654) (222222227685275403356.......1235073) (122222296956279699757.......1937669) 25148..31435 22504..08250 22612..01456 22646..10884 Group 5 - 20 mg/kg 222222245351466570952.......1024205 222222253345330233100.......2323090 222211100009999396991.......4434772 111111877677058518......678606 24135..90263 2398..27402 2062..37838 1755..70250 154-158 20 W6e-i1g6h"t -C--h--a-nge (g6r-a2m0 s) (223333329344169059285.......1883556) 32363..27757 (-311-31905870.......3912744) 224..53973S' 111189(20110464112.......0111296) 9410..50002 ------874485384108......72465600 -6147..5509S3- r1o a a Only pregnant animals were Included In group mean. Numbers within parentheses were excluded from mean calculations. b Oata analyzed following rank transformation. O O u cs Maternal Number 20736 20737 20738 20739 20740 20741 Mean S.O. 20743 20744 20745 20746 20747 20748 Mean S.D. Appendix 2 Individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 22.6 25.4 21.5 21.2 27.7 24.0 23.73 2.501 19.3 21.9 19.3 24.3 21.5 19.0 20.88 4.330 Food Consumption (gm/day)__________ Day of feestation 8 12 16 20 Group 1 - 0 mg/kg 18.8 22.6 15.1 19.2 27.0 18.1 16.9 22.2 19.5 20.2 27.4 23.1 14.2 24.6 19.8 19.4 23.6 25.0 17.4 27.6 27.8 28.5 26.7 28.3 20.13 4.132 21.55 3.597 21.10 4.145 26.05 4.284 Group 2 - 1 mg/kg 18.1 20.2 15.4 25.9 14.4 17.0 19.4 20.4 17.9 22.5 23.3 15.6 20.1 19.3 16.5 23.2 19.6 16.9 27.3 28.5 24.7 29.4 28.2 23.9 18.50 4.159 19.85 2.874 19.27 2.430 27.00 2.211 154-158 Total Food Consumption (g) Days 0-20 366.8 494.8 427.4 437.5 531.2 485.0 457.12 58.499 418.4 444.0 382.9 404.9 441.5 373.1 410.80 29.452 C04241 Maternal Number 20750 20751 20752 20753 20754 20755 20756 Mean S.D. 20757 20758 20759 20760 20761 20762 20763 Mean S.D. Appendix 2 - Continued Individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats Food Consumption (qm/day) Day of Gestation " 6----- 8 16 Group 3 - 5 mg/kg 21.5 19.4 22.2 22.6 25.6 22.7 18.3 21.3 15.4 15.4 14.2 13.7 19.0 16.3 19.2 17.5 14.7 14.5 17.0 18.3 15.0 19.0 18.2 17.0 15.8 17.0 19.4 17.2 21.76 2.385 16.47 2.735 16.60 1.880 17.66 1.269 Group 4 10 mg/kg 18.8 19.4 21.7 24.1 21.0 23.3 23.0 17.0 15.4 19.9 17.9 15.3 16.8 16.0 16.8 28.2 18.0 17.6 10.6 15.6 15.4 12.6 10.9 15.9 8.9 3.6 11.4 13.1 21.61 2.006 16.90 1.615 17.46 5.336 10.91 3.879 20 24.8 23.7 26.5 26.2 23.8 26.3 22.4 24.81 1.585 22.8 23.4 23.3 20.7 15.9 23.8 24.9 22.11 3.021 154-158 Total Food Consumption (g) Days 0-20 423.5 384.9 396.3 389.4 400.4 429.7 360.6 397.83S" 23.480 355.2 396.7 398.6 368.8 276.7 376.3 383.3 365.09s" 41.840 Maternal Number 20764 20765 20766 20767 20768 20769 20770 Mean S.D. Appendix 2 - Continued Individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 15.3 20.5 21.2 15.8 22.0 19.0 21.4 19.31 2.741 Food Consumption (gm/day) Day of Gestation 8 2 16" Group 5 - 2 0 mg/kg 1181..17 11..23 14.5 3.8 16.8 12.4 1122..14 65..88 13.3 1.9 00..62 0.2 000...063 14.13 2.743 4.74 4.028 0.32 0.240 ___ 20 154-158 Total Food Consumption (g) Days -2 i 0(1x> O o 0 N 20736 20737 20738 20739 20740 20741 Mean S.D. 20743 20744 20745 20746 20747 20748 Mean S.D. Appendix 3 Individual Terminal Body Weights, Gravid Uterine Weights , and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats 154-158 Terminal Body Weight (g) Not Pregnant 322.4 318.0 361.8 385.8 388.8 355.36 33.795 Gravid Uterine Weight (g) Group 1 - 0 mg/kg 32.9 32.8 71.3 97.7 79.4 62.82 28.982 Terminal Body Weight Minus Gravid Uterine Weight 289.5 285.2 290.5 288.1 309.4 292.54 9.634 339.0 364.8 336.6 329.0 388.2 333.0 348.43 23.213 Group 2 - 1 mg/kg 62.7 80.9 72.0 80.9 63.3 74.9 72.45 8.097 312.0 283.9 264.6 248.1 324.9 258.1 281.93 30.878 20750 20751 20752 20753 20754 20755 20756 Mean S.D. 20757 20758 20759 20760 20761 20762 20763 Mean S.D. Appendix 3 - Continued Individual Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats 154-158 Terminal Body Weight (g) 345.4 342.5 344.5 349.3 353.3 385.7 335.6 350.90 16.306 335.8 290.5 349.8 342.3 Not Pregnant 365.6 318.5 333.75 26.277 Gravid Uterine Weight (g) Group 3 - 5 mg/kg 73.0 77.6 56.1 68.8 85.0 95.5 67.0 74.71 12.846 Group 4 - 1 0 mg/kg * 72.6 61.2 65.3 86.4 92.8 70.4 74.78 12.312 Terminal Body Weight Minus Gravid Uterine Weight 272.4 264.9 288.4 280.5 268.3 290.2 268.6 276.19 10.211 263.2 229.3 284.5 255.9 272.8 248.1 258.97 19.333 004245 Maternal Number 20764 20765 20766 20767 20768 20769 20770 Appendix 3 - Continued Individual Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats 154-158 Terminal Body Weight (g) Gravid Uterine Weight (q) Group 5 - 2 0 mg/kg Terminal Body Weight Minus Gravid Uterine Weight MNautmerbneral 222222000000777777433343167908 MS.eDm. 222222000000777777444444753468 MS.eDan. 22222220000000777777755555551354620 MS.eDan. CorOpovraariLanutea ImpUlatenrtianteions N1111164673ot Pregnant 115..624 11154655 151..502 111111585564 111111146443 115..358 113..673 11111113557695 11111115232547 115..879 133..375 Appendix 4 Individual and Mean Ovarian, Uterine, and Litter Data Pilot Teratology Study of T-3551 In Rats EImffpiciaienntactyi on(Xe) ResEoraprltyions InRciedseonrcpetsion(X) Group 1 - 0 mg/kg NDeuandber of FeAtulisvees 18833087850.....25470 00021 100073.....01003 00000 11133655 7300..407 00..869 64..100 00..000 150..408 Group 2 - 1 mg/kg 889997837332......393359 001020 1600005......300004 000000 111111141454 887..721 00..854 36..360 00..000 113..722 Group 3 - 5 mg/kg 88989898290363.......3250873 0002113 120006560.......7007090 0000000 11111112432460 849..812 11..105 87..208 00..000 113..901 MorFtaeltiatyl (X) 00000.....00000 00..000 000000......000000 00..000 0000000.......0000000 00..000 154-158 ViabFieltitayl (X) 1118900062000.....70800 965..091 1111980000340000......680000 e-P'u 966..249 ' 11189890004303000.......1303000 983..208 * Data analyzed following rank transformation. 004246 MNautmerbneral 22222220000000777777766655651208937 MS.eDan. 22222220000000777777766667664578906 MS.eDan. CorOpovraariaLnutea ImpUltaenrtiantelons N111111646589ot Pregnant 116..836 11113053 1173 123..355 11111b4b3526 1111111134033 114..508 130..678 Appendix 4 - Continued Individual and Mean Ovarian, Uterine, and Litter Data Pilot Teratology Study of T-35S1 In Rats EImfpfilcainetnactyion(X) 89696223....7598 6984..44 8134..108 ResEoraprtlyions InRciedseonrcpetsion(X) Group 4 - 10 mg/kg 0001 0007....0007 01 07..07 00..532 23..968 Group 5 - 20 mg/kg Nbeuamdber of FeAtulsivees 0000 11113205 00 1127 00..000 123..428 - 154-158 MortFaeltiatyl (X) ViabFielittayl (X) 0000....0000 11190002000....3000 00..00 19020..30 00..000 937..948 ' 1 cCnJ i a Data analyzed following rank transformation. ** Ovaries were too autolyzed to count corpora lutea. 004247 004248 MNautmerbneral oNf uMmbaelers Sex ofNFuenmbearles 222222000000777777434333180679 Not Pregnant 2855 113583 MS.eDan. 42..541 63..407 222222000000777777444444753846 448997 MS.eDan. 62..382 665777 06..832 22222220000000777777755555551236405a MS.eDan. 12864757 27..508 2468885 25..394 a Sex of one pup could not be determined Appendix 5 Individual and Mean Live Fetal Data Pilot Teratology Study of T-355I In Rats PMeracleesnt Unadjusted Males Group 1 - 0 mg/kg 4463301801.....03650 44333.....13083 4112..339 03..37801 Group 2 - 1 mg/kg 536365766040......244300 333333......145245 1520..073 03..13654 Group 3 - 5 mg/kg 43837558238053.......5440099 3333333.......3844863 1574..960 03..25119 154-158 Mean Adjusted Height (grams) Unadjusted Females Adjusted 33333.....14757 03..16257 03..24489 03..04889 333333......543300 03..13066 03..22057 03..02862 3333333.......1532643 03..05939 03..13742 03..03745 * MNautmerbneral 22222220000000777777765656651279308 MS.eDan. 22222220000000777777766666678467590 oNf uMmbaelers Sex ofNFuenmbearles N1626878ot Pregnant 27..283 4577 45 51..334 MFFFFFFooooooouuuuuurnnnnnniddddddbinDDDDDDdeeeeeeaaaaaaddddddSa------crDDDDDDiaaaaaafyyyyyyice211111708849- Day 17 Appendix 5 - Continued Individual and Mean Live Fetal Data Pilot Teratology Study of T-3551 in Rats PMeracleens t Unadjusted Males Group 4 - 10 mg/kg 45656803....2404 3333....2748 6760..76 33..23 589..823 03..24538 Group 5 - 2 0 mg/kg 154-158 Mean Adjusted Height (gr ams) Unadjusted Females Adjusted 3333....2263 33..32 03..14171 03..13505 03..02892 co i o o tb iO C0 Appendix 6 Individual Visceral Findings for Each Litter Pilot Teratology Study of T-3551 In Rats 154-158 NAunmimbaelr: NNNNPuuuuevmmDmmracbbbbierleeeeanrrrriatteoonoodofffftsfffffueeeef(rtetttXeuuuut)tusssseeeeesrsesss(sswhwe)xhaiiattvamhhivn:iiinnnggedonnoornbemyeoaWrol rmillimsomoroenrite'svsvistiecsccehernraailqluveariants NPueaamrnncbeooenrmmtaaololfiieefsfsefetutusesess having one or more visceral having one or more visceral 07237 202 0.0 0 0.0 8G2073roup 1190-37 0 m^g/2k0g407 022 022 2t1 0.0 0.0 50.0 1 000 0.0 0.0 0.0 10427 022 0.0 0 0.0 32047 40247 Grou5p0427 2 - 1 mg/kg 6047 70427 28047 202 202 121 121 202 022 0.0 0.0 50.0 50.0 0.0 0.0 11 000000 0.0 0.0 0.0 0.0 0.0 0.0 7 0 ANnimimbaelr: __057 05127 G02527roup 3 053-27 5 mg/kg 0457 05527 06527 "05772-----------80527 Group905274 - 10 m00627 g/kg 02627 c0o0 1 30627 NNPNNuuuetUmvDmvImtmrenciinbbbbicssecleedeehccanorrrrueentmtlerorooioaqdapaoftfffuelllfedfffefuteeeefvvreettttuaealuuuutytrrruesssseiiedreeeaastseessssennrssttcesswwhexhaniiatatvdmhh{vien%:iiidnnng)gedotneononrsbemteyioasrWolrmillimsomoroerniet'ss 22 22 02 0.0 100.0 11 22 0 0.0 22 0 0.0 22 22 22 000 0.0 0.0 0.0 22 21 22 22 21 22 0 10010 0.0 50.0 0.0 0.0 50.0 0.0 11 o o NPuevvmrciibsseeccnreetrroaoaf llffefaaetnnutoousmmeseasaslliiheesshaav(vining) goonneeoorrmmoroere 0 000000 0.0 0.0 0.0 0.0 0.0 0.0 0.0 00 00 0 0 0.0 0.0 0.0 0.0 0.0 0.0 fO cn o Appendix 7 Individual Skeletal Findings for Each Litter Pilot Teratology Study of T-3551 In Rats 154-158 ANunmimbaelr: 70237 8G2073roup 1 920-73 0 mg/k0g2074 12047 3Z074 Z4074 Group20547 2 - 1 mg/kg 6047 Z0747 -----20847 NNPNNuuueuBvLvmmmraacoababbbPAERVSneSrergeeenretrrrxikineigeeaabttilrugrArnononovotulnaogecflffifttlleicssabafgbrffOf-fg-----trneeeeerigfea(mbdaetstttIstpihziluuuunsrs)etuny-eyauidgcssssrcnptofrooeeoecesrisitlirblmchleesmstsdsaueaeassepamtorpc-w-whte-aanckilxnhaieloriicenaepsInttvatlnlbrmhhsc--eu-eivopecnor:iItbiinhdlnnmangcascrttiiohaagseealrlupodsouncoelmslsnnorobdgIrelunran--e-oatfuaimerIlpelcncllenoseonnIaaiaodrnccomaolrorsorcrn<(lnstpmmcyocllfiloehimluem3tfomepselsnosrtoepsles--eetseridldlrtetfsesaesnlotIitehkfnseosdiak--cnesenooifeldfuemnIslients7socpae5tcinadafXlolfeitlum)ctiesoapentdleioten 23 03 04 41 24 13432 33 100 100 75 50 24 34 04 41 42 41 2 1432 3 50 25 100 75 50 75 1 2 22 31 11 1 3 111 2 1 1111 . 22 1 4123 33 2111 23 11 1 442 1 11 12 1 12 1 21 1 PNeuaarmncnbeooenmrmtaaololfiieefsfsefet0ut0usesess having one or more skeletal having one or more skeletal 00000 0.0 0.0 0.0 0.0 0.0 000000 0.0 0.0 0.0 0.0 0.0 0.0 004251 Appendix 7 - Continued Individual Skeletal Findings for Each Litter Pilot Teratology Study of T-3551 In Rats 154-158 T NAunnimbaerl: _0057 NNNNPuuuuessLmmmrnkkcabbbbPRVgeSeeeeeenrgrrrelilelcbeeiltrunvooootttlaaecgiofffflcballffOf---gfr-f-geeeefaevvestttohtsilpoItIaauuurushtu-nn-yssrrauidasscsscpsositifroneaeeaessroeliiiirlelctdfesfsssninmrueaeibiaspt7tmtecro-ps-sstww-eh5aandacinxXlhaloeriiecotaIsniII)tat(vtiinbsriXmn--hh-rseevioopcrrnsrc):iItihninlenanosoniagtleIIcrItiufohagnernmgueagrllsimosdcrucoocenmleuspssoroenbbgoroIlldliulamafgnmr-na--aaeufielmalprlrrcuecrtltpepneooennoalIilaalddlydlcyanmaraoorlesoerrrrcrtnlts(ntpeyoccpemcollilifhohylieftemuumseomeisnsssbtrocpseesietiriaifdslretfeitaslteieeodd-n 24 2 50 2 12075 2GZ057roup 3 -320755 mg/ka 43075 14 04 04 04 34 4 4 75 100 100 100 2142 2 21 4 21 22 52057 62075 34 04 14 25 100 4 4 72057 82057 G",rou*p24 9075 10 m02067g/kg____22067___ 20637 42 14 43 24 41 41 2 312 3 3 50 75 25 50 75 75 2 1 1 31 1 222 1 11 0t 1 11 121 11 1 ro C/I NAunmimbaelr: BoFAEFSnecuetxnmeestgrnreAuunadtrllreiagabrsrtnheiribmbdaaosnesred;rtnictbmhasreacclhkaeelsrlgended thoracic PNeussrnkkcbeeeenrlleet ottaaof llf f efaaetnnutoousmmseeaaslsliiehesshaavvininggononeeoorrmmoroere HumBobnere oAfligfnemtuesnets with: Fused Is c h ia and cojoined hind limbs; pubis small 00527 2 0 0.0 Appendix 7 - Continued Individual Skeletal Findings for Each Litter Pilot Teratology Study of T-3551 In Rats 51207 - 2G0527 roup 3 35207- 5 mg/kg 4057 50257 60527 1 3 4 2 2111 4 0001 0.0 0.0 0.0 25 00 0.0 0.0 1 154-158 72075 80527 Group905274" - 10 0m6027q/kg 62207 62037 2 1 11 1 000000 0.0 0.0> 0.0 0.0 0.0 0.0 1 -c o fO CC/OI - 42 - Appendix 8 References Pilot Teratology Study of T-3551 in Rats STATISTICAL METHODS/LABORATORY PROCEDURES 154-158 Bartlett, M. S., "Some Examples of Statistical Methods of Research in Agriculture and Applied Bioloqy." J. Royal Statist. Soc. Suppl., IV:137-170, 1937. Draper, N. R., and Hunter, W. G., "Transformations: Some Examples Revisited." Technometrics, 11:23-V4-.0, 1969. Dunnett, C. W., "A Multiple Comparison Procedure for Comparing Several Treatments with a Control." J. Am. Stat. Assoc., 50:1096-1121, 1955. Dunnett, C. W., "New Tables for Multiple Comparisons with a Control." Biometrics, 20:482-491, 1964. Games, P. A., and Howell, J. F., "Pairwise Multiple Comparison Procedures with Unequal N's and/or Variances: A Monte Carlo Study." J. Ed. Statist., 1:113-125,. 1976. ------- Fisher, R. A., Statistical Methods for Research Workers, Oliver & Boyd, Edinburgh, 1950. Levene, H., "Robust Tests for Equality of Variances," in I. Olkin edited, CPoanltoriAbTuttoi,on1s960t.o----P-r--obability and Statistics, Stanford University Press, Staples, R. E. and Schnell, V. L., "Refinements in Rapid Clearing Technic in the KOH-Alizarin Red S Method for Fetal Bone." Stain Technology, 39:61-63, 1964. --------------- Wilson, J. G. and Warkany, J., editors. Teratology: Principles and Techniques, The Univ. of Chicago Press, Chicago, 1965, pip. 251-277. Winer, B. J., Statistical Principles in Experimental Design, 2nd Ed., McGraw-Hill, N.Y., Chs. 3-10, 1971. r 004254