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coming HazletonInc. P.O.Box 7545 Madison,VI 53707-7545 Df,liverie3s3:01 Kinsman Blvd.,Madison,WI 608.241.4471 608-241.7227Fax 53704 OC)RNING Hazleton Sponsor: 3M St.Paul,Minnesota FINAL REPORT StudyTitle: PrimaryDermal Irritation/Coffosion StudyofT-6684 inRabbits (OECD Guidelines) Author: StevenM. Glaza Study CompletionDate: January10,1997 PerformingLaboratory: Coming HazletonInc. 3301 Kinsman Boulevard Madison,Wisconsin 53704 LaboratoryProjectIdentification: CHW61101150 Page Iof27 j-- JAN 13 1997 ........................... COM[PLL4,NCE STATEMENT PrimaryDermal Irritation/Coffosion Study.of T-6684 inRabbits (OECD Guidelines) CHW 61101150 Thisstudywas conductedinaccordancewiththeOrganisatiofnorEconon:iiCcooperation and DevelopmentPrincipleosfGood LaboratoryPracticeC,(81)30(Final). StevenM. Glaza Date Study DirecWr Acute Studies Coming HazletonInc. 2 QUALITY ASSURANCE STATEMENT CHW 61101150 Thisreporthasbeen reviewedby theQualityAssuranceUnitof CorningHazletonInc.,in accordancewiththeOrganisatiofnorEcomonic Cooperationand Development (OECD) PrincipleosfGood LaboratoryPracticeC,(81)30(Final)T.he followinginspectionwsere conductedand findingsreportedtotheStudy Directorand management. InspectionDates From To Phase Date Reportedto StidyDirector Date Reported to Manacement 11/25/96 01/08/97 11/25/96 01/08/97 Dose Preparation Data/ReportReview 11/25/96 01/08/97 11/25/96 01/08/97 RepresentativeQ,ualityAssuranceUnit Date 3 STUDY EDENTIFICATION CHW 61101150 PrimaryDermal Irritation/Corrosion Study of T-6684 inRabbits (OECD Guidelines) TestMaterial T-6684 Sponsor 3M ToxicologyService MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 St.Paul,MN 55133-3220 Sponsor'sRepresentative Roger G. Perkins 3M ToxicologyService MedicalDepartment 3M Center,Bldg.220-2E-02 P.O.Box 33220 St.Paul,MN 55133-3220 (612)736-7428 Study Director Study Location Study Timetable Study InitiatiDoante Experimental(In-lifSet)artDate In-lifEend Date ExperimentalTerminationDate Study Completion Date StevenM. Glaza Coming HazletonInc. P.O. Box 7545 Madison, WI 53707-7545 (608)241-7292 Coming HazletonInc. 3301 Kinsman Boulevard Madison,WI 53704 November 22, 1996 November 25,1996 December 9,1996 December 9,1996 January 10,1997 4 Acute Studies StevenM. Glaza Study Director Manager Steven R. Sorenson Study Coordinator JeffreyB. Hicks In-lifSeupervisor Rose M. Bridge AdministrativSeupervisor KEY PERSONNEL QualityAssurance SherryR. W. Pctsel Manager CHW 61101150 LaboratoryAnimal Medicine Cindy J.Cary,DVM Diplomate,ACLAM Supervisor ToxicologySupport Kathy Myers Manager CalvinL. Horton Supervisor 5 CONTENTS CHW 61101150 COMPLIANCE STATEMENT ................................................2............ QUALITY ASSURANCE STATEMENT .......................................3............ STUDY IDENTIFICATION ..................................................4............ KEY PERSONNEL .........................................................5............ OBJECTIVE ...............................................................8............ TEST MATERIAL ..........................................................8............ Identificati..o.n..........................................................8............ Purityand Stabilit.y.......................................................8........... Storageand Retention.....................................................8............ SafetyPrecaution.s........................................................8............ TEST SYSTEM ............................................................9............ TestAnimal ..............................................................9............ Housing.................................................................9............ Animal Diet..............................................................9............ Animal Selectionand Preparatioonf ExposureArea .............................9............ JustificatifoonrSpeciesSelectio.n..........................................1.0............ PROCEDURES ...........................................................1.0............ Preparatioonf TestMaterial...............................................1.0............ Treatment...............................................................10............ Reason forRoute of Administratio.n........................................1.0............ Observations............................................................1.0............ Termination.............................................................1.1........... StatisticAanlalyses.......................................................11............ Locationof Raw Data,Records,and FftW Report..............................I.I........... RESULTS/DISCUSSION ...................................................1.1............ SIGNATURE ..............................................................11............ REFERENCES ............................................................1.2........... 6 CHW 61101150 TABLE 1 IndividuaDlermal IrritatiSocnores.......................................1.3............ 2 AveragePrimaryDerrnalIrritatiSocnores.................................1.4............ 3 IndividuaBlody Weights(g)............................................1.4............ APPENDIX ...............................................................1.5............ ProtocolTT2071 .........................................................1.6........... ProtocolAmendment No. I ................................................2.7........... 7 OBJECTIVE CHW 61101150 The objectivoefthisstudywas to assesstherelativleevelof primaryskin irritation/corrosoifoantestmaterialon rabbitsunder sen*u'occludceodnditions.' AR proceduresused inthisstudywere incompliancewiththeAnimal WelfareAct Regulations.IntheopinionoftheSponsor and studydirectort,hestudydidnot unnecessarildyuplicataeny previouswork. Allproceduraltimespresentedinthisreport fallwithintheacceptablreangesas specifieidntheWisconsinfacilitoyf Coming HazletonInc.(CHW) StandardOperatingProcedure(SOP). TEST MATERL4L Idendficaflon The testmateriawlas identifieadsT-6684 and describedasanoff-whitleiquid. Purityand Stabft The Sponsorassumesresponsibiliftoyrpurityand stabilidteytermination(sincluding undertestconditions). Storageand Retention The testmaterialwas storedatroom temperature.Any unused testmaterialwillbe returnedtotheSponsor afterissuanceofthefinalreportaccordingto CHW SOP. SafetyPrecautions The testmaterialhandlingprocedureswere accordingto CHW SOPs and policies. 8 TEST SYSTEM CHW 61101150 Test Animal AdultalbinorabbitsoftheHra.-(NZW)SPF straiwnere procuredfrom HRP, Inc., Kalamazoo, Michigan on October30,1996. Housing Afterreceiptt,heanimalswere acclimatedfora periodofatleast7 days. During acclimationand throughoutthestudy,theanimalswere individuallhyoused instainless steelcages.Envirorumentaclontrolsfortheqnimalroom were settomaintaina temperatureof 19-to 23-C,a relativheumidityof 50% 20%, and a 12-hourlighv 12-hourdarklightincgycle.In caseswhere variationfsrom theseconditionsex;lstetdh,ey were documented and consideredto have had no adverseeffecton thestudyoutcome. Animal Diet The animalswere providedaccesstowaterad libitumand a measured amount of LaboratoryRabbitDietIHF#5326,PNE Feeds,Inc.The feedisroutinelaynalyzedby the manufacturerfornutritionaclomponents and environmentalcontaminants.Samples of thewater areperiodicallaynalyzed.There were no known contaminantsinthefeedor wateratlevelsthatcouldbe expectedtointerferweithor affectheresultosfthestudy. Animal Selectionand PreparationofExposure Area Two male and one femalehealthy,acclimatedrabbitsw,eighingfrom 2,662to 2,937 g and approximately14 to 18 weeks of age,were selectedatrandom and identifiebdy animalnumber and correspondingeartag. On theday beforetreatmentt,heback and/or fimiksof eachanimalwere clippedfreeofhairto obtainone unblemishedskinsite.The animalswere clippedas needed throughoutthestudy. 9 CHW61101150 JustificatifoonrSpeciesSelection Historicalltyh,eNew ZealandWhite albinorabbithasbeentheanimalofchoicefor evaluatingtheeffectof chemicalson theskin. PROCEDURES Preparationof TestMaterial The testmaterialwas administeredasreceived.The pH ofthetestmaterialwas notable to be determined. Treatment The undilutedtestmaterialwas appliedto theintactskinsiteon each animal'sback (approximateexposureareaof 6.25em') intheamount of 0.5mL. The areaof applicatiownas coveredwithan 8-ply2.5-cm x 2.5-cmgauze patchsecuredwithpaper tape,looselyoverwrappedwithSaran WrapP, and securedwithElastoplastt'oprovidea semiocclusivedressing. At theend of the4-hourexposureperiod,thepatcheswere removed and thetestsites were washed usingtapwaterand disposablepapertowels.The testmaterialwas removed from thetestsitesasthoroughlyaspossiblewithoutirritatitnhgeskin. Reason forRoute ofAdministration Historicalltyh,edeniud routehasbeen therouteof choicebased on themethod of Draize.2 Observations Thirtyminutesafteremoval ofthetestmaterialt,hedegreeof eraema and edema at each testsitewas read accordingto theDraizetechnique(recordedasthe4-hourscore). Subsequentexaminationswere made at24,48,72,96 hours and Days 7 and 14. The untreatedsldnof each animalwas used forcomparison. 10 CHW 61101150 Animals were weighed beforetestmaterialadministratioand atweekly intervals throughoutthestudy. Termination At terminationofthein-lifpehase,allanimalswere euthanizedand discarded. StatisticAanlalyses No statisticaanlalyseswere requiredby theprotocol. Location ofRaw Data, Records, and FinalReport The raw data,records,and an originalsignedcopy ofthefinalreportwillbe retainedin thearchivesof CHW inaccordancewithCHW SOP. RESULTS/DISCUSSION IndividualdermalirritatisocnoresarepresentedinTable 1. Average primarydermal irritatisocnoresand individuablody weightsareinTables2 and 3,respectively. Applicationof T-6684 totheskinofrabbitsunder4-hoursemioccludedconditions resultedinwell-defineedrythemaand slightomoderateedema reactions.Desquamation and fissuringwere alsoobserved.Allirritaticolnearedby theDay 14 observation. SIGNATURE StevenM. Glaza Study Director Acute Studies 0 Date CHW 61101150 REFERENCES 1. "Acute Dermal Irritation/Corrosion,O"rganisationfor Economic Cooperation and Development Guidelinesfor Testing of Chemicals, Section 4, Health Effects, November 404,ParisCedex (July17,1992). 2. Draize,J.H., "Primary Irritatioonf the Skin,"In: Appraisal of theSafetyof Chemicals inFoods, Drugs, and Cosmetics -Dermal Toxicity,Associationof Food and Drug OfficialosftheU.S.,pp.46-47(1959). 12 Table I IndividuaDlermal IrritatioSncores CHW 61101150 Animal Number Hour 4 24 48 72 96 Erythema F61197 2 1 1 1 2 F61198 2 2 2 2 2 F61199 2 2 2 2 2 Edema F61197 3 1 1 1 2 F61198 2 1 1 1 1 F61199 3 2 2 2 2 d Desquamation. f Fissuring. Day 7 14 1d 0 2f 0 1d 0 1 0 1 0 2 0 13 Table 2 Average Primary Dermal IrritatioSncores Observation Period Average Score* 4 Hour 4.7 24 Hour 3.0 48 Hour 3.0 72 Hour 3.0 96 Hour 3.7 Day 7 2.7 Day 14 0.0 The average primary dermal irritatisocnore isthetotaldermal irritatisocnoreforallthe animals(eryffiemand edema) dividedby thenumber oftestsites-(3a)teach observationperiod. CHW 61101150 Table 3 IndividualBody Weights (g) Animal Number Sex F61197 m F61198 F F61199 m Predose 2,937 2,662 2,755 Day 7 14 3,036 2,886 2,906 3,137 2,919 2,943 14 APPENDIX ProtocolT?2071 ProtocolAmendment No. I CHW 61101150 15 CHW 61101150 Sample Submiftal Form Thitsormistobeusedwhensu&n&hgswyo" formu*w acutteestinSgp.eciatlestinnegedcsanbeaas#aYrrangbeYd cmbctinfgt AcutSetudi"Depadmwtat(6082)41-7292. CHW Study No. & bM Enobsewahswnplaasndsendto.* ComingHazkdoInnc. =i KwonanBoLdward MackonW,boonsin53704 Submittbedy,--R-or,-,c^)aJ'c._ companyFullGLP Compliance:-J(- yge _ ND _ FDA (21CFR 68) EPA(rsCA--4C0FR792) DateSwnplSeent:- - Nwnber of ReportsRequired: EPA (FIFR"O CFR ISO) - OECD - MAFF mof" swmle Name: 7- (,(oQ@V -r- (,(,.P c-1 PhysicalDescription: ' SpecialHanclingPrecwAiA: Testrnaterwpuft and r,t&bnyinformatio(nkcW&V m We rtestcond 6 9 c-(, 07-7597-Y on filewithSpo toor _I- Yes - No Ted mixtuarnealysfiosrawcw*dw&anog"kYmawkY tobecwA'cto4* Y"* - bySPOnw - by CM Sample Disposal: Return toSponsor at followingaddress: wo 3rv, C-t) - ;s 5f- Dispose ofaccordingto CM SOPs SaffpW Storage Requireirmts: Rom temperature R*Otawd odw At additionalcostto Sponsor (CM willcontact Sponsor es b these addRWW durges@ Acute Oral Toxicity In Rots -TP8084 -TP3206 -TP3013 -Y.TP20-69 Llpand down LOSO procedure F14SA saom, SM-SF at5.0 gacg Corduct definedstudy I death occursat 5.0 9&g EPA screen; SM-SF at5.0 Vq Conduct &Mad study I death occurs at5.0 gft OECD memn; SM-SF at5.0 9&g Conduct definedstudy I death occurs atrLO gacg SpocW fnsUWbm: Acute Dwmal Toxicity In Robbib -TP32D7 -TP3016 - -TP2CY70 - FHSA sa"n; SM-SF at2.0g/kg EPA screem SM-SF at7-0 9/kg Conduct dermed study I death occurs at2.0 gacg OECD acmon; SM-SF at2.0 gacg Conduct defwiedstudyI death occurs at2.0 grq lalInstructions: For CH Protocol Issue Date: Study Director Only Primary skin kTroMon -TP3208 TP3014 :YTP2a7i -TP42DS -TP7145 FHSA; 6 mbbb-I abmded, I hact &Wmbbk EPA: 6 rabbks-IIntactswrabbk OECD; 3 rabbb-I ktact sit@trabbk DOT corfowvkr.6 tabbks-llrtactsitafmbbk Rwftax"r. 6 mbl*s-2 hftctsk"ftbbk (one aft*with LNA mcposure) Special Instruction: Primary Eye Irritation -TP6360 -TP3209 -TP2012 -TP3015 -X-TP2072 -- Low-volunw pmewwre; 6 rabbb unwashed FHSA: 6 fabbb unwashed 1978 EPA; 6 rabbb urwashed, 3 washed 1982 EPA*.6 mbbks unwashed OECD. 3 mbbks unwashed 3 rabbitswashed at4 second& 3 rabbb washed at30 sawrds Specialkwrurtbm- Guln*a Pig Sensitization -TP2017 -TP6164.EC -TP2008 -TP62BO EPA MagnLmwwlGigman maximization OECMC Magnussun-Kigman maxknizawn Buohlw sensitization PhotoaDergenic contact dormatkis(ArmsUong) Speciallnstnx:tbu: White copy-CHW Yellow copy-SubryfMor 16 '"HAZLC-ICC:N W 15 C 0 N S I N P0 ST MADISO...0IS1CO1..1.C. E a 0S31?1.71-I1.4..1 CHW 61101150 C@@ UbwatwV S@ Col.WW SRonsor: 3M St. Paul,Minnesota PROTOCOL TP2071 Study Title: Primary Dermal Irritation/CorrosionStudy in Rabbits (OECD Guidelines) June 1, 1993 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProlect Identification: HWI (.(Io 17 CHW 61101150 TP2071 Page 2 STUDY IDENTIFICATION Primary Dermal Irritation/Co"osionStudy in Rabbits (OECD Guidelines) HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Final Report Date C,, (See sample submittalform) 314 Toxicology Services 220-2E-02 3H Center St. Paul, MN 55144 John L. Butenhaff, PhD 3M Toxicology Services 220-2E-02 3H Center St. Paul, MN 95144 (612)-733-1962 Steven M. Glaza Hazleton Wisconsin',Inc. P.O. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. BuildingNo. 3 3802 Packers Avenue Madison, Wl 53704 Week of (1-2!C-CtcWeek of Week of I- a7 -"'#7 18 CHW 61101150 TP2071 Page 3 1. Uuft PrimaryDemal Irritation/CorrosionStudy in Rabbits (OECD Guidelines) 2. Puripose To assess the relative level primaryskin irritation/corrosioonf a test material on rabbits unodfer semioccludedconditions 3. Regulatory Compliance This study will be conducted in accordancewith the followingGood Laboratory Practice Regulations/Standards/Guidelines: Conduct as a Nonregulated Study 21 CFR 58 (FDA) 40 CFR 160 (EPA-FIFRA) 40 CFR 792 (EPA-TSCA) C(81)30(Final)(OECD) NotificationNo. 3850, August 10, 1984 (JapaneseMAFF) Notification No. 313, Narch 31, 1982, and as amended by NotificationNo. 870, October 5, 1988 (JapaneseMOHW) All procedures in this protocol are iin compliancewith the Animal WelfareAct Regulations. In the opin on of the Sponsor and study director,the study does not unnecessarilyduplicate any previous work. 4. OualityAssura= For regulatedstudies,the protocol,studyconduct,and the final reportwill be audited by the Quality AssuranceUnit in accordance with Hazleton Wisconsin (HWI) Standard Operating Procedures (SOPS) and policies. 5. Test Material A. Identification (See sample submittalform) B. Physical Description (See sample sutxnittalform) C. Purityand Stability The Sponsor assumes responsibilityfor purity and stability determinations(includingunder test conditions). Samples of testmaterial/vehiclmeixture(s)(if applicable)for concentration,solubility,homogeneity,a,nd stabilityanalyses will be taken before administrationif requestedby the Sponsor. These samples (if taken)will be sent to the Sponsor afterexperimentalterminationfor possibleanalysis. 19 CHW 61101150 TP2071 Page 4 D. Storage (See sample submittal form) E. Reserve Samples Studies of less than 4 weeks in experimentalduration will not have reserve samples retained. Reservesample(s)of each batch/lotof test materialwill be taken if this study is more than 4 weeks in experimental duration. The test material reserve sample will be stored at HWI in a freezer set to maintain a temperatureof below O*C for 10 years per HWI SOP. The Sponsor will be contactedafter 10 years for disposition in accordancewith the appropriate.regulatoryGood LaboratoryPractices. F. Retention Any unused test material will be discardedafter issuance of the final report,unless directedotherwiseby the Sponsor. G. Safety Precautions As required by HWI SOPs and policies 6. ExRerimental Desicin A. Animals' (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/Hazleton Research Products, Inc. (3)Age at Initiation Adult (4) Weight at Initiation 2.0 to 3.5 kg (5) Number and Sex 3 of any sex (6) Identification Individualnumbered ear tag. 20 CHW 61101150 TP2071 Page 5 (7) Husbandry (a) Housing individually,in screen-bottomstainlesssteelcages (heavygauge). (b) LQgd A measured amount of High Fiber Rabbit Chowe 15326 (PurinaMills,Inc.). The food is routinelyanalyzed by the manufacturer for nutritionalcomponents and environmental contaminants. (c)Water Ad libitum from an automatic system. Samples of the water are analyzed by HWI for total dissolved solids, hardness,and specified microbiologicalcontent and for selected elements, heavy metals, organophosphates, and chlorinated hydrocarbons. (d) Contaminants There are no known contaminants in the food or water that would interferewith this study. (e) Environment Environmentalcontrols for the animal room will be set to maintain a temperatureof 19 to 23*C, a relative humidity of 50% t20%, and a 12-hour light/12-hourdark cycle. (f)Acclimation At least 7 days (8) Selection of Test Animals Based on health and body weight according to HWI SOPS. An adequate number of extra animals will be purchased so that no animal in obviouslypoor health is placed on test. (9) Justificationfor Species Selection Historically,the New Zealand White albino rabbit has been the animal of choice for evaluatingthe effectof chemicals on the skin. 21 CHW 61101150 TP2071 Page 6 B. Dose Administration (1) Prepa ation of ExRosure Area On the day before test material administration,the back and/or flanks of each animalwill be clippedfree of hair. The treatmentsiteswill be inspectedfor interfering lesions,irritation,or defectsthat would,@reclude the use of any of the animals. The animals wi be clipped as needed throughout the study. (2) Dose Administration Before administrationof liquid test materialsthe pH of the materialwill be determined(if possible). The test materialwill be appliedto the intacttest area (approximately6.25 cm2) on each rabbit, in the amount of 0.5 mL in the case of liquidsand 0.5 g in the case of solids. If a liquid,the testmaterialwill be applied undiluted. If a solid,the testmaterialwill be moistenedwith 0.9% saline. Aerosol materialswill be discharged into a beaker and administeredas a liquid. The area of applicationwill be coveredwith a 2.5-cm x 2.5-cm gauze :atch secured with j)apertape and overwrapped with Saran Wr p* and Elastoplast tape to provide a semiocclusivedressing. The rabbitswill not be collared during the 4-hour applicationperiod. (3) Reason for Route of Administration Historically,the dermal route has been the route of choice based on the method of Draize. (4) Removal of Test Material After the 4 hours of exposure, the patches and test material will be removed as thoroughlyas possible using water and/or an appropriatesolventwithout irritatingthe skin. C. Observation of Animals (1) Readingof Dermal Irritation Approximately30 minutes after removing patches, the degree of erythema and edema will be evaluated according to the Draize technique (Attachme!nt1) and recorded as th6 4-hour score. The intact skin of each animalwill serve as its own control. Subsequentreadingswill be taken at approximately24, 48, and 72 hours after patch removal. If irritationis present at the 72-hour examination, additionalobservationswill be made at approximately96 hours and at Days 7, 14, and 21, or until all irritation 22 CHW 61101150 TP2071 Page 7 has cleared. Basedon the levelof irritationobservedat any of these time points,the study may be terminatedat the directionof the studydirector. (2)Body Weights Just beforetest materialadministrationand weekly thereafter(whenapplicable). D. Pathology Any animalsdying during the study will be subjectedto an abbreviatedgross necropsyexaminationand all abnormalities will be.recorded.After necropsy,the animalswill be discardedand no tissueswill be saved. At temination.ofthe experimentalphase,survivinganimalswill be designatedto be sacrificedand discarded. E. StatisticaAlnalyses No statisticalanalysesare required. 7. Report A finalreportincludingthose items1-istebdelowwill be submitted: Descriptionof the testmaterial Descriptionof the test system Procedures Dates of experimentalinitiationand termination Tabulationof irritatiodnata Descriptionof any toxic effectsotherthan dermalirritation 8. Locationof Raw Data. Records,and Final Report Originaldata,or copiesthereof,will be availableat HWI to facilitateauditingthe studyduringits progressand before acceptanceof the finalreport. Whenthe finalreportis completed, all original.papedrata, includingthose itemslistedbelowwill be retainedin the archivesof NWI accordingto HWI SOP. Protocol and protocol amendments Dose preparationrecords In-life-records Body weights Dose administration Observations Anatomicalpathologyrecords(if applicable) Study correspondence Finalreport(originalsignedcopy) 23 CHW 61101150 TP2071 Page 8 The followingsupportingrecordswill be retainedat HWI but will not be archivedwith the study data. Animal,receipt/acclimationrecords Water analysis records Animal room temperature and humidity records Refrigeratorand freezer temperature records 24 CHW 61101150 PROTOCOL APPROVAL TP2071 Page 9 gee,., John L. Butenhoff, PhD Sponsor'sRepresentative 3H Steven M. Glaza Study Director Acute Toxicology HazletonWisconsin, Inc. Representative Quality AssuranceUnit HazletonWisconsin,Inc. (TP2071.3M) Date/ Date 61- /9 Date 25 CHW 61101150 TP2071 Page 10 Attachment 1 PrimaryDermal IrritationScoringScale (DraizeTechnique) (1)Erythema and Eschar Fomation No erythema 0 Very slighterythema(barelyperceptible) I Well-definederythema 2 W erate to severe erythema 3 Severeerythema(beetredness)to slighteschar formation(injuriesin depth) 4 Highest possible erythema scare 4 (2)Edema Fomation No edema 0 Very slightedema (barelyperceptible) I Slight edema (edgesare well definedby definiteraising) 2 Moderateedema (raisedapproximatelyI mn) 3 Severe edema (raisedapproximatelyI m and extendingbeyond area of exposure) 4 Highest possible edema score 4 26 CHW 61101150 CHW No. PROTOCOL AMENDMENTS Coist@ flccnl Amendment No. I Effective Portionof ProtocolBeing Modified: Applicablesectionsof the protocol. Reason for Modification: To identify the location where the study will be conducted and to reflect a company name change from Hazleton Wisconsin, Inc. (HWI) to Corning Hazleton Inc.-(CHW). replace wherever ai)plicablethe following changes Modification: Corning Hazleton Inc. (CHWI 3301 Kinsman Boulevard. Madison. Wl 53704 (G21/01-07-91) Study Director Approval: 27