Document rn58z99BJEQQd3gG5KKO7jaE

1# BIQAYAE,ABJJIY_QJEXBAD,ERQM SOILS: PHASE I. PARTS I & II; DOSING METHODS AND ANIMAL MODEL SpJvla^mt4^e_.Rftg^ar.gn..flontgac^,tot\feen Louisi.m^-Sfea^^J.QJtyegsifcY. ,,jSchOQJL Q Medlclpe in Hew Orleans (LSPSMNol s E>IDu Pont de Nemours & Company, Inc, fDU PONT> I.C. OBftL. iX>XXCmJ,-,,aftrJaj^^ imrX^JGS^WTTH LEAD SURGICAL RESEARCH PROJECT NO. 101 RBQiSCQIi Louisiana Stats University School of Medicine in Hew Orleans IACUC Approval # 90S N40673 SUBCHRON1C_.QBAL TOXICITY: 30-DAV FEEDING STUDY IN RATS & MINI* lSg_Jil32L^ES SURGICAL RESEARCH PROJECT WO. 101 TABLE OF CONTENTS INTRODUCTION. . OBJECTIVE ................ SPONSOR AND TEST FACILITY .................................... STUDY DESIGN. .............. MATERIALS AND METHODS .......... A. Test Material . . B. Test Species. . . C. Animal Husbandry. D. Pretest Period. . E. Assignment To Groups. ...... F. Diet Preparation and Analyses . . G. Body Heights. . * * * . ..................... H< Food consumption and Intake of Test Material I. Clinical Observations and Mortality J. Feces Collection and Analyses . . K. Lead Analysis........................ L. Pathological Examination. ... STATISTICAL ANALYSES. .......... SAFETY AND HOUSEKEEPING ......... RECORDS AND SAMPLE RETENTION. ...... REFERENCES. APPENDIX I - Animal Identification Marking System (AIMS) APPENDIX II - study Personnel and study Dates a p p e n d ix III - Technical Data on Mini-pigs APPENDIX IV - Vitae of Investigators 3 3 4 4 5 5 5 5 6 7 7 8 8 8 8 9 9 10 10 11 11 DUP050055253 SI rxc ORAL TOXICITY: 30-DAY FEEDING STUDY INRATS & MINIjmcrPXG.s-.wm .ram S^BgICAL..,.RESEAPGH PROJECT. NO* W ABSTBACT: Multimedia exposure of children to lend is recognized as a health problem of international proportions. Ingestion of soil lead and dust incidental to hand to south activity presents one of the principal direct pathways for exposure to non-dietary lead in areas with significant soil contamination Environmental lead contamination is derived from a variety of sources includings leadbased housepaint, auto emissions, smelter emissions, wind-blotm tailings or mining wastes which have been used for fill in residential developments and/or land-fills. This phase of study seeks to determine which animal model and dose level provide the best measure of soil-lead bioavailability, transport from the intestinal lumen to the blood stream. o w t bc t xvss The overall objectives of this study aret 1) to gain a more thorough understanding of the chemical and physical characteristics of soil that influence lead bioavailability and 2) to develop scientifically and legally defensible determinations of soil lead bioavailability in site soils to support the development of sitespecific cleanup goals. The proposed research uses soils with a wide range of lead concentrations, test feeds animals a diet mixed with the soil, and assesses lead bioavailability using biomarkers. As currently conceived this study will proceed in three related phases. The objectives of this first phase are the determination of critical aspects of study design (e.g. animal model selection and concentration effects) that will be used to tailor the definitive protocol for the second and third phases involving site soils from various locations throughout the DU PONT corporation . This first phase of study will evaluate the bioavailability of lead In soil when incorporated into nutritionally adequate purified AIN 76 basal diet and fed to male rats (Harlan Sprague Pawley and male mini-pigs (Charles River Yucatan Hinipig YU) for 30 days. The oral route of administration was selected because it is the potential route of human exposure. DUP050055254 SPONSOR ftlID TEST FACILITY This study is sponsored by E.I. du Pont de Nemours & Co.* Xnc., Wilmington* Delaware. The study will be conducted in the Department of Surgery* Louisiana State University, School of Medicine in New Orleans, in accordance with all applicable Good Laboratory Practice (GLP) standards (40 CFR, Part 792). STUDY DESIGN The experimental design is as follows: Task l (Concentration Effects and Lead Acetate vs. Lead Nitrate for Control Lead Normalisation Study: holds soil constant, varies Pb Concentration and Pb salt form) C -O IL-t x. o 1 S o 1 1 + + Pb-Soll Group PMcQ Group Group B>r.g.o,U # Rata/-Gxoup # Rats/ Group # Ra.t.s/Group. & Soil Pb Cone . 15 4 10* 75 10 5 13 5 25 55 85 11 5 14 5 3 5 <30 ppm (Conti 65 -375 ppm 95 -750 ppm 12 5 -1500 ppm IS 5 -3000 ppm Task 2 (Animal Model Comparison Study; holds pb constant S -1000 ppm, varies animal model) Group Group Mini-* 0 BatB,/G,roup._ # pjqs/SEPUB Pietary i2sgp.fintr-ati.oh. Ip p bX 16 10 IS 5 17 5 <30 ppm soil + PbACO at 1000 ppm 19 5 1000 ppm * 5 rats in groups 4 & 16 and the mini-pigs in group 17 will be bled at 3, 7, 14, 21, 28 and 30 days; Feeding and testing may continue, if blood lead levels do not plateau; this will require contract amendment to provide for additional animal care & testing per diem costs. -s file schedule for study functions is as follows: Function Soil Lead Concentration Diet preparation Diet Sampling Clinical Sign Observation Body weights Food Consumption Feces Collection Feces Analysis Blood lead Levels Approximate Study Dav Pretest 15 30 Analyzed - Analyzed <- Once----------------------------------------------------> ab <--------------------- Daily - Daily ---------------------> - Daily-----------------------------> <-----:------- weekly ------------------------> <- c -- c ----- c ---- C ----c-C-> a Sampled for homogeneity and concentration at start of study b sampled from feeders for concentration analysis during study c Orbital bleeds on Croups 4 & 16 (rats) and venipuncture bleeds on Group 17 (mini-pigs) at 3, 7, 14, 21, 28 & 30 days; cardiac blood from all other animals at sacrifice, 30 days. MATERIALS AMD METHODS A. Test Material The test material will be supplied by the sponsor, DU PONT* The test material will be assigned a LSU-Surgery fast Material Number (LSU-Surg TM #) and LSOSMNO will provide a chain of custody for the test material. The concentration of lead in the diet/soil mixtures will be confirmed by analyses at the beginning and end of the study, during diet mixing and from feeders during the study. B* feat Species One hundred fifteen (115) male Sprague Dawley rats, approximately 21 days of age, with body weights ranging from 40 to 65 grams will be acquired from Harlan Sprague Dawley, Indianapolis, IN. Rodents will be delivered in Harlan climate-controlled trucks to the LSU-Surgery laboratory once a week. The Hsd: SD rat has been selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. Eleven (11) male Yucatan mini-pigs, approximately 30 days of age, with body Weights ranging from 3 to 6 kilograms will be acquired from Charles River Laboratories, Wilmington, MA. Mini pigs will be crated and shipped to New Orleans by air freight. The weanling Charles River MINIPIG YU has been selected on the basis of its low body weight and its suitability as a model of soil-lead ingestion by children and with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. DUP050055256 -6- C. Animal Husbandry Each rat will be housed, singly in filter-top polycarbonate plastic cages to prevent test animal exposure to environmental lead and with cage cleaning and feeding conducted in a laminar-air-flow hood, to protect laboratory personnel from lead exposure. Raisedwire mesh floors will be used to prevent coprophagy. HEPA- filtered inflow-outflow filter-top caging systems (jCfcffiE&n, Allentown or Lab Products). which can be adjusted to maintain positive or negative cage air pressures, will be utilized to contain lead spiked diets and to protect the test animals from environmental exposure to lead. Individual cage racks will be relocated within the room weekly and cages on the racks will be repositioned every 2 weeks* Each minipig will be housed singly in air-tight isolation cages (Plas Labs) to prevent test animals exposure to environmental lead and with daily cage cleaning, feeding, diet & animal weighing conducted in a laminar-air-flow station (Standard Safety Equipment Co.. Palatine, IL hood, to protect laboratory personnel and other test animals from lead exposure. Raised-floors will be used to ?revent coprophagy and aid in feces collection. HEPA-fiitered nflow-outflow filter-top caging systems, which can be adjusted to maintain positive or negative cage air pressures, will be utilized to contain lead spiked diets and to protect the test animals from environmental exposure to lead. Individual cage racks will be relocated within the room daily, Animal rooms will be targeted at a temperature of 23 2C (rats) and 25 5C {mini-pigs. See Appendix III) and relative humidity of 50 25%. Animal rooms will be artificially illuminated (fluorescent light) on a 12-hour light/dark cycle. All rats and mini-pigs will be provided test diets and deionized water ad libitum. During the pretest period (see section D), all rats will be fed Teklad 5% basal diet and all mini-pigs the Ziegler 5% basal diet. During the test period, all rats and minipigs will be fed. the diet of their respective treatment group (see Section P) hSU-Surgery Laboratory has an animal health monitoring program which consists of periodic food and water analyses for contaminants and sampling freshly washed cages and cage racks for bacteria. This program is monitored and administered by the LSUSMNO laboratory animal veterinarian. This program will be applied to this study. D` Pretest Period Upon arrival at LSU-Surgery Laboratory, all rats and mini-pigs will be removed from shipping cartons and housed three per cage (rats) one per cage (mini-pigs), in a quarantine room. The rats and mini-pigs will be: DUP050055257 -7 o quarantined for a minimum of six days, o identified permanently with a subcutaneous microchip? . American Veterinary Identification Devices, Inc. (AVfp), and cage identification; o weighed daily; and, o observed with respect to weight gain and any gross signs of disease or injury. The rats and mini-pigs will be released from quarantine by the laboratory animal veterinarian or her designee on the bases of body weights and clinical signs of all rats and mini-pigs at the end of the pretest period. Rats and mini-pigs that die or are euthanized in extremis during the pretest period will be necropsied to check for the presence of disease. Dependent upon these findings, further diagnostic procedures will be employed at the discretion of the study director or the laboratory animal veterinarian. S* Assicmmontjto^roupa Animals will be assigned during the pre-test period separately for each task. Animals will be assigned to the groups specified in the Study Design. Each rat and minipig will be housed individually, assigned its own LSU-surgery animal number/ and permanently identified with a microchip placed under the skin (see Appendix 1}. Rats and mini-pigs that have not been assigned to a test group may, at the discretion of the study director, be released for use in other studies, or be sacrificed by sodium pentobarbital overdose (loo mg/kg body weight) and discarded without pathological evaluation. Diet, Preparation and Analyses During the test period, rats and mini-pigs in each group (as identified in the study Design) will be fed the following purified diets: a) RATS - IBKLAD 5% Basal Diet (T5%BD) and b) MINI-MINI-PIGS - ZIEGLER 5% Basal Diet (Z5%BD). These diets will be custom formulated by their respective diet manufacturers so that each diet contains complete normal nutrition for each species when the diets will be diluted (Weight/weight) with test soil containing <30, -375, -750, -1500, or -3000 ppm lead. LEAD/soil will be added to T5%BD & ZS%BD and thoroughly mixed for a period of time that is adequate to assure homogeneous distribution in the diet. Control diets will be mixed for the same period of time. All diets will be prepared once and refrigerated until used. DUP050055258 - Before the beginning of the study, samples will be collected from each concentration of diet prepared with l e a d /soil. These samples will be analyzed to verify concentration and homogeneity in the test diets. Homogeneity samples will be collected from the top, middle, and bottom of the diet mixer. samples will be frozen until analyzed. Homogeneity diet samples may also be collected if a new method of diet preparation is used. Also, if a new method of diet preparation is used, it will be documented in the study records. Stability testing of the soils will not be performed. USEPA publication SW-846, Methods for the Analysis of solid Waste, specifies a holding time for solid material samples of six months with proper refrigeration for samples being analyzed for lead, since lead in the samples to be tested in this project is hot expected to degrade during the study period, stability testing is not deemed to be necessary. Extra diet samples may be taken at the discretion of the study director. Whenever extra diet samples are taken, a sample of control diet will be collected and frozen the same day as diet preparation. Once during the study, samples will be collected from randomly selected feed jars to verify concentration of lead in the feeders. Samples will be collected from every group. Backup samples will be taken on the same day from other feeders in the same group and analyzed if needed. Feeder samples and backup samples will be frozen on the day of collection. 0. jLoj3y--y.elght.s All rats and mini-pigs will be weighed once a day unless experimental findings warrant a more infrequent weighing schedule. H. FO-04_Consumption and Intake of Teat Material The amount of food consumed by each individual test animal each day will be determined daily throughout the study. From these determinations and body weight data, average test animal daily food consumption and intake of test material will be calculated. I* gasAL-Ool-le.otion and Analyses Feces Will be collected daily as follows: rats & mini-pigs entire fecal output for each test animal will collected and weighed (wet weight), it is expected that the entire rat fecal output (3 to 10 grams) will be collected daily, weighed, and pooled for 7 days. From this pooled sample, total weekly lead excretion will be determined. Mini-pigs will have their fecal output (0.5 to 1.5 kilograms) collected daily, weighed and a representative 25 gram DUP050055259 sample collected for lead analysis* J. clinical observations and Mortality Cage-site examinations to detect moribund or dead rats and mini-pigs and abnormal behavior and appearance among rats and minipigs will be conducted once daily throughout the study. Moribund rats and mini-pigs will be euthanized. Moribund and dead rats and mini-pigs will be given a pathological examination. At every weighing, each rat and mini-pig will be individually handled and examined for abnormal behavior and appearance, jc. Eeafl analyse? samples of soil, diets, and tissues will be analyzed for total lead content according to the schedule specified in the Study Design. All rats and mini-pigs that are found dead, accidentally killed, or sacrificed in extremis before the end of the study will be necropsied. After approximately 30 days of continuous feeding, surviving rats and mini-pigs will be sacrificed by mathoxyflurane and exsanguination, and necropsied. The order of sacrifice will be random among all treatment groups. The following tissues will be collected and preserved in formalin from rats and mini-pigs that are found dead, accidentally killed (tissue integrity permitting), sacrificed in extremis, and from those rats and mini-pigs sacrificed by design after 30 days on tests Stomach Liver Pancreas Small Intestine (duodenum, jejunum, and ileum) Large intestine (cecum, colon) Kidney Brain Peripheral nerve (sciatic and/or vagus) *Bone (femur) Muscle (thigh) All gross lesions Portion collected for lead analyses Microscopic evaluation of these tissues will be performed if warranted by experimental findings. Blood will be collected as indicated in the Study Design and during exsanguination at the final sacrifice. Whole blood will be DUP050055260 analysed for lead content. 10 - STATISTICAL ANALYSES Body weights, body weight gains, food consumption, will be analysed by a one-way analysis of variance, When the test for differences among test group means (the value of the F test statistic) is significant pairwise comparisons between test and control groups will be made using the Dunnett's. test. Significance will be judged at alpha 0.05, Other methods will be used- if appropriate, at the time of analyses. Relative bioavailability will be determined by comparing tissue lead levels from the soil test groups with tissue lead levels from their respective lead acetate and nitrate control groups. SAFETY AND HOUSEKEEPING Lead in soil has a low acute oral toxicity. Good housekeeping procedures will be practiced to avoid contamination of diet preparation facilities and potential health hazards. To avoid skin contact, gloves will be worn when handling either the test material or diets. In addition, the test material will be handled in a chemical hood. Diets will be prepared in properly ventilated areas. RECORDS AND SAMPLE RETENTION All original records will be retained at LSU-Surgery Laboratory or at the E I. du Pont de Nemours and company, Wilmington, Delaware. Preserved wet tissues, paraffin blocks, histological slides, and be retained at LSU-Surgery Laboratory. A portion of test sample will be collected for archive purposes prior to the outset of the study and retained at LSU-Surgery Laboratory. Any change, addition or deletion to the protocol will be documented as a Protocol Amendment and a copy sent to the sponsor. REFERENCES Allcroft, R. 1950. Lead as a nutritional hazard to farm live-stock. IV. Distribution of lead in the tissues of bovines after ingestion of various lead compounds, j. comp. Path, 60:190-208. Ball, D.J., G. Argentieri, R. Krause, M. Lipinski, R.L. Robison, R.E. Stoll, and G.E. Visscher. 1991. Evaluation of a microchip implant system used for animal identification in rats. Lab. Animal DUP050055261 Sci. 41(2):185-186. --11 Barltrop, 0. and Meek, F. 1975. Absorption of different lead compounds. Postgrad. Ned. J. 51:805-809. Barltrop, D. and Meek, F. 1979. Effect of particle size on lead absorption from the gut. Arch. Environ. Health 34:280-285. Carlson, A.J. and A. Woefel. 1913. The solubility of white lead in human gastric juice, and its bearing on the hygiene of the lead industries. Am. J. Public Health 3:755-769. Chaney, R.L., H.W. Mielke, and S.B. Sterrett. 1989. Speciation, mobility, and bioavailability of soil lead. [Proc. Intern. Conf. Lead in Soils: issues and Guidelines. B.E. Davies & B.G Hixson (eds.)] . Environ. Geochem. Health 11 (Supplement):105-129. Chidambaram, M.V., M.B. Reddy, J.L. Thompson, and G.W. Bates. 1989. In vitro studies of iron bioavailability: Probing concentration and oxidation-reduction reactivity of pinto bean iron with ferrous chromogens. Biol. Trace Element Res. 19:25-40. Deere, J.c., and TerHaar, G.L. 1977. Lead levels in tissues from rats fed soils containing lead. Arch. Environ. Contarn. Toxicol. 6:111-119. Day, J.P., Fergusson, J.E. and Chee, T.M. 1979. solubility and potential toxicity of lead in urban street dust. Bull. Environ. Contam. Toxicol. 23:497-502. Dodds, w, j. 1982. The pig model for biomedical research. FASEB 41(2):247-256 Elfving, D.C., Hasqhek, W.M., stehn, R.A., Bache, C.A. and bisk, D.J. 1978. Heavy metal residues in plants cultivated on and in small mammals indigenous to old orchard soils. Arch. Environ. Health 33:95-99. Freeman, G.B., J.D. Johnson, S.C. Liao, P. I. Fader, J.M. Killinger, R.L. Chaney, and P.D. Bergstrom. Effect of soil dose on bioavailability of lead from mining waste soil in rats, Chem. Spec, and Bioavailability 3(3/4):121-128. Harter, R.D. 1983. Effect of soil pH on adsorption of lead, copper, zinc, and nickel. Soil Sci. Soc. Am, J. 47:47-51, Hassett, J.J. 1974. Capacity of selected Illinois soils to remove lead from aqueous solutions, commun. Sol. sci, plant Anal. 5:499-505. Healy, M.A., P.G, Harrison, M. As lam, S.S, Davis and C.C. Wilson. 1982. Lead sulphide and traditional preparations. Routes for DUP050055262 -a?,- ingestion, and solubility and reactions in gastric fluid. J. Clin. Hosp. Pharm. 7:169-173. Hemphill, C.P., M.V. Ruby, B.D. Beck, A. Davis and P.D. Bergstrom. 1991. The bioavailability of lead in mining wastes: Physica1/chemical considerations, Cham. Spec, and Bioavailability 3(3/4): 135-148. Klaassen, C.D. and D.W. Shoeman. 1975. Biliary excretion of lead in rats, rabbits and dogs. Tox. Appl. Pharm. 29:434-446. baVelle, J.M., R.H, Poppenga, B.J. Thacker, J.P. Giesy, C. Weis, R Othoudt, and C Vandervoort. 1991. Bioavailability of lead in mining wastes: An oral intubation study in young swine. Chem. Spec, and Bioavailability 3 (3/4) :i05 -111. Mathur, S.P., and Rayment, A.F. 1977. The influence of trace element fertilization on the decomposition rate and phosphatase activity of a Hesic Fibrisol. Can. J. Soil Sci. 57:397-408. Mielke, H.W. and J.B. Heneghan. 1991. Selected chemical and physical properties of soils and gut physiological processes that influence lead bioavailability. Chem. Spec, and Bioavailability 3(3/4):129-134. Hiller, D.D., B.R. Schricker, R.R. Rasmussen, and p. Van campen. 1981. An in vitro method for estimation of iron availability from meals. Am. J. Clin. Nutr. 34:2248-2256. Killer, E.R. and D.E. Dllrey. 1987. The pig as a model for human nutrition. Ann. Rev. Nutr. 7:361-382. Mushak, P. 1991. Gastro-intestina1 absorption of lead in children and adults: Overview of biological and biophysico-ehemical aspects. Chem. Spec, and Bioavailability 3(3/4);87-104. Kylroie, A.A., Koore, L., Olyai, B. and Anderson, H. 1978. Increased susceptibility to lead toxicity in rats fed semi-purified diets* Environ. Res. 15:57-64. Reddy, M.B., E.J. Browder, and G.W. Bates. 1988. Cannulated swine and in vitro approaches to iron bioavailability, pp 173-185. In A.S. Prasad (ed.). Essential and Toxic Trace Elements in Human Health and Disease. A.R. Lies, Inc., New York. Schricker, B.R., D.D. Miller, R.R. Rasmussen, and D. Van Campen 1981. A comparison of in vivo and in vitro methods for determining availability of iron from meals. Am. J. Clin. 34:2257-2263. Stara, J., Moore, W., Richards, M., Barkely, N., Neiheisel, S. and Bridbord, K. 1973. Environmentally bound lead. III. Effects of source on blood and tissue levels in rats. Pp 28-29 In EPA DUP050055263 Environmental Health Effects Research Series A-670/1-73-036, Steele, J.J., B.D. Heck, B.L. Murphy and H.S. Strauss, 1990, Assessing the contribution front lead in mining wastes to blood lead. Teg. Toxicol. Pharmacol. 11:158-190. Weis, C.P. and J.M. LaVelle. 1991, characteristics to consider when choosing an animal model for the study of lead bioavailability. Cheat. Spec, and Bioavailability 3 (3/4): 113-119. Wise, A. 1981. Protective action of calcium phytate against acute lead toxicity in mice. Bull. Environ, contam. Toxicol. 27:630-633. Woefel, A., and A.J, Carlson. 1914* The solubility of lead sulphide ores and of lead sulphide in human gastric juice. J. Pharmacol. Exper. Ther. 5:549-552. Zimdahl, R.L. and R.K. Skogerboe. 1977. Behavior of lead in soil. Environ* SCi. Technol. ll:1202-1207. DUP050055264 -..14.- EBfiS&Cflfr ftPMPyx X SURGICAL RESEARCH, -PROJECT NO. 101 ANIMAL IDENTIFICATION MARKING SYSTEM Rate will be identified by a subcutaneously implanted microchip MC#. The numbering system will be according to the following scheme: Task 1 P o .n t g-o, 1__.Kj s l AJ, + + Pb-Soil Kale PbAcO Male Pb (NOj) 2 Male Pb-Soil Rats MC# . Rats MC# Rats . MC# i 101-S 2 201-5 4 111-10 5 211-5 7 121-5 8 221-5 to 131-5 11 231-5 13 141-5 14 241-5 3 301-5 6 311-5 9 321-5 12 331-5 15 341-5 Acetate, Nitrate & Soil Pb Cone. <30 ppm (Control) -375 ppm -750 ppm -1500 ppm -3000 ppm Task 2 (Animal Model Comparison Study) Male Rats 16 1$ MC# 501-10 6 01-5 Male Minisim MC# 17 560-65 19 660-65 --Dietary, Cpnceptra.tasn,. (BPJO. <30 ppm Soil + PbAcO at 1000 ppm 1000 ppm DUP050055265 =..15..= EBOTSgot APPPNPIS U SURGICAL RESEARCH PROJECT NO. 101 Study Personnel and Study Dates gtufly.Funotion Study Personnel Study Director LSUSMNO James B. Heneghan, Ph.D. Professor, Surgery & Physiology Consultant Xavier Howard Mielke, Ph.D. Associate Professor, Pharmacy Clinical Pathology LSUSMNO Charmaine Foltz, D.V,M. Lab Animal Veterinarian Analytical Chemistry LSUSMNO cecelia Breaux, B.s., (M.s.) Laboratory Supervisor Study_Dates Proposed Project Start August 1, 1992 Phase l: Task 1, Concentration & Control Lead Normalization Study in Rats Pretest October 26, 1992 30-Day November 25, 1992 Scheduled Euthanasia November 25, 1992 Phase It Task 2, Animal Models (Rats vs. Mini-pigs) High and Dow Doses Pretest February 15, 1993 30-Day March 17, 1993 Scheduled Euthanasia March 17, 1993 Proposed Report Dates for DU PONT/LSU-Surgery Studies: Task 1 Task 2 February 1, 1993 April 5, 1993 DUP050055266 1&. SUBCHRONIC .ORAL TOXICITY; 30-PAY FEEDING STUDY IN RATS & MINI-* MINI-PIGS KITH LEAD SURGICAL RESEARCH PROJECT NO. 101 mtggg.il Date Janes B. Heneghan, Ph.D, (LStlSMNO) Study Director <> LSUSMNO Institutional Representative O.U.-FQMC cc. T. S. Bingman R* C. Shea M. S. Bogdanffy P. D. falling DU p q n t Institutional Representative LSUSMNO J. Patrick O'Leary Marilyn L. Zimny Ronald Smith Larry McDermott DUP050055267