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SECt1iITY CLASSIFICATION OF THIS PAGE ("aen Date F.nrared)REPORT DOCUMENTATION PAGEGOVT ACCESSION2.REPORT NUMBERI READ INSTRUCTIONSBEFORE COMPLETING FORMNO.TITLE (and Subtitle)4,3Fate and Distribution ofMycotoxin in Guinea Pigs7.3.RECIPIENT'S CATALOG NUMMERS.TYPE OF REPORTPERIOD COVERED5InterimH-Labeled T-2AUTHOR(a)6.PERFORMING ORG. REPORT NUMBERB.CONTRACT OR GRANT NUMBER(-)M. R. Watts, E. P. Burrows, R. E.Matson, E. C. Hauer, and R. W.Wannemacher, Jr.G. E10.ADDRESSUS Army Medical Research Institute of InfectiousDiseases, SGRD-UISDetrick,Fort871-AA12.REPORT DATE3 AugustUS14.Army MedicalMONITORING& DevelopmentResearchCommand13.21Office)from Controllingry'V NAME & AODRESS(If dlforet-"15.AD-A 144 759DISTRIBUTION16.Distribution unlimited - Approved1I.SUPPLEMENTARYNUMBER OF PAGES 8 FiguresSECURITY CLASS.(.fDECLASSIFICATIONSCHEDULEthia report)DOWNGRADINGfor public releaseBlock 20, if different from Report)InSTATEMENT (of the abetract enteredDISTRIBUTION15.1984Report)STATEMENT (of thli17.TASK21701AND ADDRESSCONTROLLING OFFICE NAMElt.MDFrederick,PROGRAM ELEMENT. PROJECT.AREA & WORK UNIT NUMBERSCD,NOTESTo be published in19.-Toxicology and AppliedKEY WORDS (Continue on ,eveeae"7-2 mycotoxin,9ST-PAc'r rccitri'-2toxinis aIf necessary-Idedistribution,ue. are!si avt,&land Identify by block number)metabolism,f rnl.eopotent cytotoxiýPharmacologyilaPy nml@tdfrlfguinea pigs,TLCby block number)metaboliteproducedby theYusariIm1)peCiesThe fate and distribution of Hl-labeled T-2 toxin were examined in maleRadioactivity was detected in all body tissues within 30 minlguinea pigs.dose (1.04mg/kg) of T-? toxin.The plasmaafteran Im injection of an LDinit ialwith intration cirve of radioachVvtty versgus time was mull /2,E)half-IltFabsorptionBile contained a large amount of radioactivitveliminatt,-n (TI/2,A) was t.8 hr.Do1.73COJT1-OOconcen-1O4V615 ISov OII OLETESECURITYCLASSIrICATIOM OF THIS PAGE eW hnatr.FM-t,.f.d
SECURITY CLASSIPICATION OF THIS PAGE(3rh., Data Zngtr*M7which was identified as HT-2,4-deacetylneosolaniol, 3'hydroxy HT-2, 3'hydro yThese T-2 metabolites are exT-2 triol, and several more-polar unknowns.creted from liver via bile into the intestine. Within 5 days, 75% of the totalradioactivity was excreted in urine and feces at a ratio of 4 to 1. Theappearance of radioactivity in the excreta was biphasic (T1/2,A 2.2 hr, 1.5days and 8.2 hr, 1.7 days, for urine and feces, respectively).fletabolic derivatives of T-2 excreted in urine were T-2 tetraol, 4-deacetylneosolaniol, 3'hydroxy HT-2, and several unknowns.These studies showed a rapid appearance inand subsequent logs of radioactivity from tissues and body fluids.However,The disdetectable in tissues at 28'days.radioactivity (10 dpm) was stilltribution patterns and excretion rates suggest that liver and kidney are theprincipal organs of detoxication and excretion of T-2 toxin and its metabolites.SECURITY CLASSIFICATION OF THIS PAGEO'hwn Date Enrered)
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Best Available CopyF'atv!5.5 .-t tCý ace3 , .w.atts J ,:. it, aed'unningTtl4e:zn 0iF08te3H -L a-te 'ed.-E. F .T- ! CO QtuQ'-, r:w--4 . eurer:a ns,,,H-T-2R;e-e.,;.E.1 .1 n.D:ntl.ra -,e. a,cner .,'ne aI .San .i:,-2a. t sor .e*, 2n4t07inA l.rbtl 840824077
3zate anc Distribution ofWatts,J.G.C.and Wannemacher.T-2ton:inisM. R.,H-LabeledDinterman.R.R. E.,soe:.ies.:ate and distributc,e',-aained:n male ouinea pigs.C.,E.inwas detectedlarge amountaBile containedunknowns.into the intestine.excretedhr,metabolitesWithin 5 days,1.7 days,inand severalsubsequentfor urine and feces.These studiesloss of radioactivityradioactivitydistributionthe principalmetabolites.(1,43dom)was stillorgans ofdeto::icationrates.ia bilei-verwasThe aooearanceo41.5 days iol.showed a rapijdetectablepatterns and excretionfrom(TS,2 .A 2.2 hr.frcm tissuesaooearanceand body fluids.intissuessuggestand excretionas HT-2,and severalradioactivityrespectively).urine were T-2 tetraol,unknowns.1.1.8 hr.was identifiedare excretedthatme'e less2.,(T,/ 2 .9) was75% of the totalthe excreta was biphasicinof T-2 excretedHT-2.r-:mal/l.c)versus titiT-2 triol,'hvdroxv:n urine and fe-:es at a ratio of 4 toradioacti.,ityi.'TheseTI-whichof radioactivity:'hydroxv HT-2,A-deacetylneosolaniol.more-ooiarabsorot or halbodyal!(.,o4radioactcurve ofhaei-l:ze of eliminationThe initial6 min.thanE.by the Fusar.umorcducedRaalcactxvitywIt h an -nitialwas muItahascHauer.[ 3 HI-laleled T-2 tc.n ., weren ofThe olasma concentrat-on. n.7-2P.,Pharmacol.ti sues within :.0 min after an im injection of an LDsO dosec;Face.Guinea Pigs.B-rrcws.Ao l.Toticol.metabolitec tant cvtotoxicaMatson.(1984)Jr.W.,inT-2 MycotoxinandHowever.at 28 days.liverinTheand kidnev are:: T-: toxinand its
Best Availlable Copyto',in.T-'IP ,5-dia cetom vc eefE:1zl.aa'-I:it or zi-e*e:a. :TI-Omcsonb-".s.4haveta-:in to T-2 tet-raci,a!.9 7e:19@.*7!;K8I.that-2:.T-Z is m.etaboi:.:edtoýUeno ettoan". am j sever aI% etabali :e theand several ur rc trs-.necisclanicl.hýIndrox:cor)ren er si .e st*.J.v st,'e,Chi eron:w4-HT-:. These studies snowecualor lesser tox-icitya.thesanoles are ofter. obtained ats uch cr tniis reacson we :-cnduztec a.etabc!i -n3z 7-.Ies:ned t"? itscct::in . I, str , Lti cr.rdia# ca :t i.,itv af#t erin T-. to:.:intlhe 4:rst 49--Z hr Pe-'.cd aiter "me aomin:strat cn c4i t2e .:re o::I'ts 11,r oc ia eknýetatciitez,cellsis raoidi: , etabo i :ed. :-e.,icus studies vere7n cases oi hmann toxicoses.the quiinea oig.lceus5ineoc a1an i'2b-n emetabolized into products Cf:zincerned %itha n!n the lactatring cow'h'varo% v T-2. andiecause T-: tc:ýi C i.metaticlis m o:r. t!me4-deacetylnecso! anic!I qSO):animal' . whi le t-o: ler chi ic-ensYosh:: aa et a!.,deacet ilnecsol an, ci.to I m.et z. sin t:Sui s ietF pc--,el. ecortecan!1 rats -eta cc! 1::e T---un, -icns .Matsumoto et a:.a 7.,!4;92anz, act,:ea latioratzr es a,naUe n17eco.i-4'l-trichotnecenear-joizr remacher.r ce e7is a ocitentI3C.tnte I3nost o- te 1a.reIThese :2oaournds are c.ýtctc.:-c to cuit,.red:cnocuS.aeo-tc e :n SiLhvldro:, .-:sa tc.,:ic metabolite oroduced b-i the F'4;arzuqeo3.th te*:-ene)oe- es7- Aeth Ibut vrylo;: v --SO-over :a da-;s.Eoeci-icallv. wemetabc!: sip. and e!cr-et: an oatternsa s ing 1e i .t i i)ec t ion o4f14 1]1abe! ed T-: to::in 1in17m so doing we hoped to determine which oi the najordetec-ted in bocy 4iu:ds. cou d be ise- as ciiaindicators
intoxication.of trichotheceneMETHODSStandards for TLCspectrometrv(GC/MS).chromatoqraphy-massand T-2 tetraol)and by qas(TLC)was determined to be 99% by thin laver chromatographytrici,FurityT-1 toxin was purchased from Calbiochem-Behrings.ea.snts.from Calbiochem.were purchasedT-2 toxin waslateled with tritium in the C-Z position by New Enoland Nuclear'purity,specific activity 8.9 Ci/mmolusino(97% radiochemicalThe purit'1977 ).the method :2 Wallace et al.,'HT-2. T-(T-2,was determined by HPLC asof 9T-2)well as TLC. .im injection of T-2 mycotoxininjected with 1.0,2.5,1.75,in the guinea pig, si:: ;A-! ,i.nea1.5,to::in in ethanol:glycerol:watermg/kg(mean *I- SEM)To establish the toxicity of aninnguinea pigs.of T-2 sycotox1.25,or 0.5 mg/kg of T-20.75,1.0,The LD9 0 was 1.04 1-(2:3:5).with a mean time to death ofcias were0.2516.6 hr (Pace et al,,1993)'.Treatment of Animals and Sample Collection.male guinea pigsweighing between 450 and 500'(LDso)g,were injected[3H]-T-2 toxin per ml in ethanol:glycerol:water(Pace et al.,1993).im7 ma of T-2 toxin and 200 uCi ofwith 0.5 ml/kg of a solution containingnon-exchangableOne hundred and twenty fivestudy,In the first(2::5)twelve toxin treatedguinea pigs were housed in stainless steel metaboi ccages,water intake were measured over the 28 day post-exoisureani food andtime oeriod.Urineand feces were collected daily from those guinea pigs that survived theIn the second study,toxin challenge.guinea pigs were housed in groups of1 2 each and allowed food and water ad libitum.killed at 0.5,3,6,12 hr: 1, 2,Z, 7, 14,4Six guinea pigs wereand 28 days.Guinea pigs were
selected at random for necropsy for histopathologicalpi0s 4ere anesthetized with 0.5 mL-(;g of aRo-'oung,:c2from the ga2lictedTissue saamples,includingstomach,coliectec,weiched and7e"e tnat:n ri a.tab.,radIo ct .it.of plasma.ivliver,urine,irine ccllecteakidnev,heart,k i,v. frozen between iiuitz -.aci tD asma.iatI:randBile wasfrC.the tladoe,.I½na. soieen,large and small intestinesa 1iouiuomuscle,d nitradrenals.and fat pad were-co-ed clamps.;rine and bi7e were ana 1'zed forcounter'nnaiuotand bile was analyzed oy TLC to separate and identify theto.in n-tab;!ites.nbladder andThe guineaYixture of Ketaminelood was remo',ed b., cardiac cuncture.and.testes, brain,I:analyses.Fr-coated s lcathk ? were deve:ooed us I.ngel TLC plats(2e21cn.2.Ft!ssolvent svstems:two secuLi nta.o. r r.orm: ethyl acetate: ethanol.50: 25: t al.198).Mycotcxnstandards were visualized based on a chromogenic reaction between 4-opnttrobenz ,l1 pvri'dine and the 12-!3-epoxy group(Takitanieta!.,1970)and samples were scanned for radioactivity with a Bioscan BIDl00radicisotopic scanner".The radioactive zones then were scraped,extracted with boiling ethyl acetate, and filtered throuch a class woolplIug.The filtrates were evaporate under a stream of nitrogen.Theresidues were derivatized with I drop each of methylene chloride andtriluoroaceti.anhydride,sealed,and alIOwed to stand at room temperature!.!-: hr before nitrogen removal oi the :iouidsThe residues we-edss oi.:ed in a m nimum of acetone and an alyed ,mmediateiv0Hewlett Packard 5 0812equipped wth a 2'm X ,.2 mm ID fused silicacapillary column (cross-linked OV-!.the source.by GUiMS usino a!I um thick)Sour:e temperature was 20 0 C,interfaced directl., toand GC conditions were st Available Cor)N,
tq e ,,1600 C,,. V:1: ' 2.0OV / iiM to:5,6-eElectron-elti.nc.a more rua.I i nqý/l i70O.,Distribustion.to determine the tissue distribution ofý radioactivity,awe:ghed sample was oxidi: Ad3 and the radioactv tv determined in aliquid scintillation counter.Metatalites have not yet been determined intissues.-kinetic Analysis.The absorption and elimination of T-2 toxin weredescribed by means of kinetic parameters.K and; Ka,in the followingequat. on:Cr A(-e-4AT e-IT)where C? concentration in plasma at time T, A constant, K.absorption constant,Ks elimination constant.ccn:entration against time is(Karlcg et a&.,1978). The semilog plot of thea straight line with Kr as the slopeThe concentration curve i-s divided into absorptionand elimination phases with half-liveselimination constants as follows:related to the absorption andTI,12.A !n2/K.and T1 2 .a!n2/Kc.Statistical analysis.The coefficient of variation, C s/mean,to describe the amount of variationin the data.was usedThe standard deviation(s)expressed as a percentage of the mean was less than or equal to 15%.RESULTSGeneral Observations.In guinea pigs receiving an LD a dose of T-2 toxin,mnarkedly reduced but slowly returned to normal by day 7.food intake wasThis reduction infood intake combined with the intoxication resulted in an average loss ofbody weight of 73 g within 3 days.iBy 3 hr post-exposure,&the guinea pigs.r.
describead pE-t Y de N-.c.:la et ai.sabsort.ha -lecretahr t. g5;.:nset).1,from blood2 . 2 hr v. ITh7 :umulative excretioninaxis).of JletabaThe bile,act ,.-ty(5x !0domrnites in Siie! and cturalwas still7'.5 days.4:1(Fig 2,injection ofwhile in feces,theAt 29 daysdetectable in the excreta.and Urine.obtained from the gall bladder,(Tatleof the totalpeaked in urine 24 hr afterover the firstradioactivity slowly increasedradioactivity71%the urine and feces at a ratio ofand rapidly decreased over the next 4 days;the tc;n4n a1Ysithe arine and feces of theIn a five day period,Radioactivitywitn itshr)of radoacti witv inshown in Figure 2.radioac-ivity was excretedleft veo-ical., tt-.nurine-correlated-S.2 hr,ndin fe-es. Te appearance of labelcuinea cig is--d,.,oacti,The acoearanze c;with initial absoroption hal;-lives of :.2 hr in urineas biphasicdiisaoorrance-2.1.anationmnin and an initlless thanon na f-lie .witn anwasinset.ultiophasi:,i,T e plasma :oncentrat.ci curve.nsiailar toand sorphclogical changes wereHze-ntolcica!:20C V. 12 h-.th-osetheir temperatures reached a nadir ofto develop hypothermia;starte:12 hr ithad the highest specificcontained17% of the total administeredThe biliary metabolites were identified bvof a TLC plate",illustratedin Figure 4,assignments were made on the basis of electronspectra of their GC-separatedtrifluoroacetate derivatives.andimpact massStandards(T-.,
HT-2and T-2 tetraol)unknown samples.zpectralOther structuralibrary prov:ded4-t'eacet vInecsolanlal42were run.underug/m!,aI.,(9 ugiml).1991)occurinassignments were based on the massby Mirochaand Pawlosky(Yoshizawa et al.,gmii'a.orng with HT-:19921(X.5the same conditicrs: as. werethe'i41980)5:3),Thus.Dehydration ofthe 3'-hydrotylfrom 7'-hydroxy T-2 triol83 isisdue to the conjugated2/z 200 are characteristicepoxytrichothec-97eneaddition,a substantialrelated polarinvestigation.The base peak at si:and the fragments above(Paolosky et al.,ug/ml)was found,Figure 6 isurinary metabolites of T-2.(1.46, 12.trichothecene-under structuralwere T-2 tetraolto 7ug/ml),radioactive peak located at(Z.6 to 6 ug/ml),3 hydroxy HT-2 (3 to 4 ug/ml)( 4.7 to 7 Mg/ml).4-andmetabolitesAfter treatmentthat remained at the origin with p-glucuronidase,twoless-polar radiolabeled peaks were smnarated by HPLC.suggests that atof theThethe origin mas composed of severaland comprised 5-20% of the total measured radioactivity.additionalIn24, 71 hr the major urinarymore-polar trichothecene metabolitesthe material1984).of unidentifiedand is12,13-a representative radiochromatogramAt 3,identified by GC/MS,deacetylneosolaniolseveralresultingBy 24 hr the concentrations of all biliary metabolites haddecreased 4-fold.neWabolites,that of the 2'-isomerof r moiety at C-S.quantitymetabolite(s)(Pawloskv et a!.,and S*-unsaturateddepicted in Figure S.trifluoroacetatestometabclites was deducedmass spectra of mixtures of the respective 2'A typical mass spectrum,Cole ethas been shownconditionsthe oresence of the 3 -hydro:yseparated by GC.T-Z (Yosn::awa et aI.and 3'hydroxv T-2 trial areadily under the trifluoroacetvlationSc94:.was the major metabolitegmi;, 7'hydroaxva ratio of(personal communicatonI.Thisleast one of the components of this peak was conjugated asS8Bof
mmOmwM4lucuronide.At a!:E'ý 26 dars T-2 tetraoltime ooints thne urinarvScetectcn.!in :tsTRecover,and boneiof ied T-2 metabolite.of unmetaboiizedT-2 was bel2w.'o::ie at 12 nr,t-ewas the onlyadministered radioactivtvcourts. ranged-om 74-l:r)Y.:argeand in the3 hrs,7irstntest'ne :-4 hrstwc phases:ad-iinisteredradioactivitvof tne totalr-aoactivity remained associated witn the G!and 3 show the distributionthe next 24 hr.of labelradioactivity in most tissues appeared at 70 min,Whilethe totalkidney (TableI).the specificThe specificactivity,of the totalless than 1%.i.res 7tract.The Peakthen raoidly declined.counts associated with the kidneythose with the 1iver,a racidAfter day 7,into major tissues.(dpmrmgFig.r7)were less thanof tissue)activities of the heart,was higher inbrain and testeswere amcng the lowest measured.Muscleand fatThe values fordeposit51Fig.S)containedthe percentage of radiolabelthe highest total counts.:n muscle and fat areaor: ximates based on muscle being 40% of the total body weiohtbeingI!% of the total body weight.comparable tthatfound in heartand tatThe speriflc activity in muscle was!Table!,.DISCUSSIONRadiolabeledT-. toxin was distributed in,nFfollowed by a slow increase to: erýskinPaj:oact.ivtv cea'edintestine atappeared in the largein thee.,cludino carcass.all tissues within 30 min-
an'af.ter,The rapi&a injection" of, thr toxin.reationff labelsuggests that toxic effects could begin shortly after, exposure to the to::in.Z;.cept in the large ,ntestine and bile,rap:dly declined,the radiolabel peaked b, -0 min andwith no measurable !0.og-ter2 ac:umlatIon.the ear v 4-ane (12-24 hr)in genera!.uIstribution patterns paralleled the districuticns"-ecorted using chicken 'Chi et a!.,1173; Yoshi:awa et a!.,qS0swine (Robison et al,,1979 ) models,1 andwith a slightly higher concentrationattributable to bile and the G! tract.2ur study represents a cumprehensiveinvestigation cf tha Jiitributionand metabolism of T-2 toxin past the 72 hr time point.The plasmaconcentration curve shows that radioactivity can be detected in plasma asearlI as 6 min and as late as 28 days aiter exoosure to the toxin. Even at:;ýe earliesttime pcint (10 min),(detection limit,metabolite o-lug/m!),we were unable tc detect unmetatolized T-suggesting that the long-term toxic element isT-2, possibly HT-2.aThe eliminat:3n phase of the plasmaconcentration curve appeared to be multiphasic due to the presence of toxinisetabolites.The distribution and excretion patterns suggest that thz liver ismajor organ for metabolism and detoxication of the toxin.liver rapidly metabolized T-2 to HT-2 which,,netabolites,theBy 30 min, thealong with several more-polarwas eliminated from the liver via the bile.This issupportedby the fact that the specific activity of the bile was highest at every tinepo.nt.The bile, therefore,plays animportant role in toxin elimination.Sinca bile was collected at death and not continuously, we could notdetermiae how much radioactivity actually passed through the liver and bile.However,liver perfusion studiesa single pass through the liver,is ext-acted,(Pace and Watts,1983) suggest that during70% of the total administered radioactivityand 50% appears in the bile.10
ýeperendent.a ,e.-e.:3e,-:t'.e*'ers:24-:I ez't::ircIen.tericthatesuzhi:nradzoacti.vitv' etraol,be agoodtheunknowntotairapidlym:nto HI-2etofand polarin this:es,-satuniesbenetit23 day stjd.,,c.;.T. 2 tetraciet:n;nat ewasal.urine maythe toxin.the urine and bile revealedaccountedmetabolicwhichisprofilethenthe oresencefor apprcx-matelvsuggestsconverted40-56"that T-2 to::into one ofofofisthreeas follows:presented hereregardingitesa!sTe -bethan T-2 t-:;the quantitatlanr-rThese7n Urine.The majoressmetabolites AnichThe4 or(Matsumoto: i -ma,o'!er thedfnd.ol'2:.as charcoa.,12.t metatbohist:zathci C.iNeue.i9the ;eces.toofandto: inhithT-2 tetraolvia 4-deacetylneosolaniol tYoshi:awa et3'hydroxv HT-2 (Yoshizawa et a!., 1992)Phydroxv T-2 triolv.a T-2 triolor 7'hvdro:.v HT-2The datatetrao!T-2metabolinedHT-2 HT-1HT-2insu.--estSrec -tede uldIof exposureTLC and ,Cz,"Sseveralaet?::oaedc ty afsuggestindexiadministeredt.The dataI?81).pexcretedwasthe::davs!or thek.rennec etrad oactivit., trnm tileThe slnw elmination,. -e'4 T-:orally(sbsorbant" dinacti--azv.t.'sinj;a-ecesaz:ount:te3i Pea'.a*in.c4 -odentstrictt r.stu jvt.o*itest;rne tght 'L:tnL.ýei2t-rcqress onsuggestmetabolitesthe metabolismofthat furtherinurine willcharacterizationyield valuableofa!.,19SQ)the T-2informationT-2 tc:xin.study we investigatedthe absorption,11distribution,metabolism,
'771-ý21:I;e4 i I Z-.tf'.t I.iietatolites.t. I it I an34T-:te-incon - ert@ý4Tte Jetectiot:Jaminist-Atar.ýe z§jsed !2% ýtsindits71-: tociwaWitesvititct slag fpolar tetatclitessuggestspraiucts.I n 4u I ne a, 213%.týSt'OngInt .3'.4r%.neastor* eizects!znq:4T-27110 quinea*a to Dc! aras :5 3.&,si-!2a*
'lee.1W?C t :ýacc -. tn.e:.arezi m t t 9etneu'zarnc Lzthle -are:'e';e:t t-e C: -an! ,e:*.azcra!tr.-ed;tatio an c4ý f.4fnlA:&cmcrr-epo ndernce snould be addressed.,rma.med, cal icer.;,neer ing Researcntcran.d Devel opienta.hlaratorv.En; Iand Nuc 1ear . Boston , Mass.'21e2. a,.er Lokh art,Q MarkOhioT San Diego, Cal i.!aleStrain:-teuci berq1:'!.tieratzr ,An, &a: Care.Ca ,:cr'em-Be".rin.tb , the aevr, can rosszc: attnt, es are 4 .: ted11,eaI17.: ttltte C4 LaCor 3t 3rv Ana a I resources, 4atiznal ýesear:h Counc;1.*ic:4i33ýawinee.'All Bci.txiaticnPre-ccated silica gel1521Lab A'nimals"arsasCounter.thinCSearlenaviInc. Des Plaii'es3.laver plates. Ell Science, Cincinnati ,'I!
.4 -- Piescn.!!C.a h gto4 c '. ie: :* rI"loop * C.
-EFERENCE ,*M0;Zr.*E\:ECE. .-.i,/cotc i:nsNImOLA, AFL 012.D,]NER.W.R.H.ZOX,ýram0.B.tuar : uMATSEU3TO.41.REBAR,A.14.,ANC CARLTON,NIEL3EN.'4. .TO.P,T.FvoijN.AND FASMULSEN.UENO,Y.(107B).ae t,Abstractý2445,7?1-40-2.CUTLER.B.M.,Vat . Prý.-j.J.:srad oactivity from tritusPharmac,,l.CUNFER,e eter Ls corum.c cr-i s in the o uir ea o ia .0.Pr44.,::4-170.W. W. (197. T-'zTox. c11l9S7.STUART.H. G.trich.thecene. The isolation and identification oi several-.itvte J.totin in chicks. 7T:ýo!. Appl. .LE1J.r4c: zr%i. :atm1cc: e4,ects and.Excretion and tissue distribution ofiabeIed T--leM.z-PCeracministraticn.routes c4!!,t-aoeri tz ree3.1973).ND NEU E:D. ::* - 4r.-.To;:To-icoalogcaltoxinCQc :nnet:cs.approachjs. ;c ;,L4r:-to the
oetabo!-tes :t Cisiar.a.tadr.:,es.W. i.:an* .A. Mehles., eas.,Cesseti."AAH4TTERJ72E. .v.ar'ae in Sautheastnizlo;i:3lof T-2 taxim in Rice.\f!I. Fate and distribut:iatAsia.J.t.As:.4.F.tt-?WJSTSN.Oft. Atal.-Y ES,S.W.Ches. 66,1485-:4 .3. SCE. :.,rat livers.PACE,3. 6.,*TTS,WAM.10 83).Metabo!ism of T-2 in isolated periusedFed.Proc. 42,1809 (Abstract Z05).WATTS.M. R.,AND MATSON,C. F.oc T 2 toxin in guinea pigs. Fed. Proc.PAWLOSKY. R. W.(soeers)M!ROCHA.C. J.,(19Se).Fate and distribution42, 6Zb (Abstract 1957)AND YOSHIZAVA,T. (1984jof two metabolic derivatives of T-Z toxin (TC-!Reaction productsand TC-O)when"-ea:ted with triflue-oacetic aco anhvdride. submitted to the 3. 4;r:c.RG?!SnCN.ZHI.7.1.S.S.,'1971). ; F, dvChen.S,;TO,:n!M!R0C!CA. C.J.,H. J. EEifRE4S,I.C. WEAVER,--G.A.,Distritution af tritius-labeled T-: toxin in swine.27,. AND OHTSUSO,MvcotoxinsKURTZ,(I.F.H.j.1411-141".K. (1974).Trichothecone toxins of Fusarius hivale.Purchase,ed.),p.I I279--91,Elsevier Scientific
rrEc-:4. .ND WANNEý!CHEF,'ýiMPz4 ar:teipns,.rt esis'. bst -actUENO,Ana 1eaJ 9oY.,G0,TSUBO,1,'7.OSHIZAWA.ketaoa!xc.2)Studies,ra : r172.i-i T-: i1i:tion':, 1tzre :e;is.Road r icks,FarýS Ltr.44e 1' .9' Y.,AMAKA,Fusaria. .pn.tc:::, c aIcanimal,fateZ.T-2 toxinJ.,in3 ndoed s .anftmycoto.x ns:e4.inaENOMOTO,.ectofM. ANDsc rieneInhi tcna oroteinrS5-296.74,T-2BEHRENS,SAITO, M.,SATO,N., AND SHIMADA. N. (1 ?7-GC)A , C. j.of radi olabeledMIROCHA,of3 .V. . 4!ýen man,u, an40. :3-3a.1SHI!.i.UoeaM.,skin-necrct ed.,trchotheceneceils.FATHIPE,Snthes .sT.,ofE.xp.k.an d Mf. ,mycIcqgy, chesistr',NA.AJIMA ,study onJ.:Ato nS-:4.k.,Comparativen !!I!.Scut.,3. 7;e:r.ss. . W. Hes se It ,n e,FcrestNAýA.J!MA, M. , SAKAI,ina 4dr-107-0.Kverea4 1WALLACE , E. !.W.R. W., 2R-.Tri cathecene m'ycator.ns,iSHTAWA,Comparat:tesynthesisa t ra;a.era.!ic :.r:ý.-metabolites ofUENOi.V.Pathoto: .: :ý:ý.-a av .UENO,tissueTr'c. Potreckense.st.S:-27.Y.int :.-,Y.t.Iz77UENO ,ae.:.ae.RUto::n.JJ.a lactatingC.,cow.I SON.T. S. . ANZ FENTON . s.4or:.FioAND SWANSON,CheI.25.S. P.(1981).Foyd Corset. Tovi:ui.19,
S7-indý17Zj ,4ND M!POCHA.C. j.-4T-: toy. ns : New meta!:oli tez- Zý T-: t-nrViSH41.AWA. T.T.;fNO.?*,LO!.SWA NSON.S. P.,AND MIOCAC. 2.198;1 1:.-iT-:eioiein the e-,c-eta c brciler ch::kens administered 3u-labeied T-Z totin.39.s
LEGENDScl:.te eniicinset li.trates-t L e z cse.:c-eton in ur ne V0,,teasti-ein tke zoacaerits 3-:ur.,e :ortcta: cun.41at, ,e eý:cr-et,r. All ana Cal m.and fecesand small- itestine.n; ext adýmnistrat on ofdata obtained from 6vayof lateled T-2afterOistribjtion of radioactivitv in the 31 tract (stomach,.t ead,1nnstrat onc; railoactivit,i ,n oi:lasma co'cent-atona ,!ult1-Znai:C.E,creti,--igureof the concentratM)and ble 40) of guinea jigs after aEach point is the aoeraqe ofaioeleo 7-2 to::zr.guinea pics:A), largeThe coe'f :ient of variation was lessthan 15.F igure 4.Representative radiochromatogram of bile collected from the gallbladder of guinea pigs administered labeled T-2 toxin.WT-2,T-2 trial and T-2 tetrad!)re . :2Lre .radiochromatoqramlabeled T-2 toxin.T-2 tetraol)are indicated above the scan.T-2 trio!.of urine from guinea pigsTLC standardsadmitistered7.(T-2,are indicated above the scan.Mass zpectruea of the clefin from 7 'h-dro:. Representat-veTLC standards( T-2.HT-2.T-2 triol,andDistribution of radioactivity in liver (). kidney (0) and heart. 4fguinea oigs after a single im administration of labeled T-2 to,.in.Each coint is the average of data obtained from 6 guinea pigs.Figure S.Distribution of radioactivity in muscle 4)and fat.0'pigs after a singie im administration of labeled T-2 toxin.the average of data obtained from 6 guinea nigs.19of,uineaEach point is
'able 1.Specifi: radioactivity in tissues of guinea pi gs. after a single imadministration of labeed T-2 toxinSeecifipe radioactiviiTimeTissues0.5 ev Plasma"81tec",*ean "'7124Z,.TC,.1S:7.' 77.31.642.514.031.36.Z7.323.7538values ior sý,x guinea pigs.than 15%.met weighat!(hr)3 141.3134.0t1U1l12, .5I,!. a74.95 0.30.30.40.20.73.6The coefficient of variation was lessT-2 was administered at a dose ofof plasma is.S.2.36 x 100 dp2/kg t dy weight.OSpecific activity168expressed in units of dpm/ml.activity of bile is expressed inunits of dpm/w1.I mg/kg or0.4".1).o2.40.1.14.30.61.0
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AD-A 144 759 15. DECLASSIFICATION DOWNGRADING SCHEDULE 16. DISTRIBUTION STATEMENT (of thli Report) Distribution unlimited - Approved for public release 17. DISTRIBUTION STATEMENT (of the abetract entered In Block 20, if different from Report) CD, 1I. SUPPLEMENTARY NOTES -T
