Document 7Rpq4JRpD1kKzdJkvVG2waZBj

Role of Polychlorinated Dibenzofuran in Yusho (PCB Poisoning) " TAKASHI KASHIMOTO, Ph.O. HIDEAKI MIYATA, Ph.D. SHIN) I KUNITA, PtkD. Osaka Prefecture! Institute of Public Health 1-3-69, Nakamlcht, Hlpshinari-ku Oala, Japan TA-CHENG TUNC, M.D., FtuD. SHU.TAO HSU, M.D,, Ph.D. KING-1 EN CHANG, MJ>. SHU-YING TANG, M.D. Department of Biochemistry National Taiwan Unfvmfty College of Medicine 1 )ervAl Road Taipei, Taiwan GEN OHI, M.D., Ph,D. J UNICHI NAKAGAWA SHUN-ICHI YAMAMOTO, M.O, Ph.D. Department of Hygiene and Preventive Medicine Faculty of Medicine Unhranity of Tokyo 7-3-1, Homo, Bunkyo ku Tokyo 113, Japan ABSTRACT. In tha Mood of IS patients with Ytohe or "polychlorinated Mphanyl poisoning" that occurred In 1979 In Taiwan, wit found polychiorinaiad dfoenzofurwo (14 of IS) and polychlorinated quaiarphawyli (IS of IS), at wall at polychlorinated biphenyl* (IS of IS). Tha moan concentration ratio of thaaa substance* woa apprexImatNy 1: 160 :30(L Based on the following eridenee, we propoae that polychlorinated quattrphonyii wore major pathogenic / substances In tha devMopmtnt of Yuaho: (1) Qlnical manh f fnuiioru and course of Yusho patients an tBiproportion- v at tty severe and panbtent for tha oheerved Mood Iavail of polychlorinated Mphanyli, whDq padanta who wart occtn pationally exposed to pure potychtorinaiad Mphonylt aka characteristically mHd and benign clinical court* despite polychlorinated Mphenyl levels often much higher than there noted in Yusho patients; (2) Polychlorinated debontefurans show a marked tendency to accumulau In the liver,- which might explain frequent presence of laundice and other abdominal symptoms In Yusho, which are, apin, not observed In those with occupational polychlorinated biphenyl poisoning; (3) Toxicity of polychlorinated diban. xofurens Is a hundred to ten thousand daws great* dtan But of polychlorinated biphenyls and polychlorinated quatorphanyls In animal experiments. "YUSHO" it a poisoning characterized clinically by acneIlka dermal Ittiortt and variety of dbnninitional symptoms. In 1961, an outbreak occurred in western pins of Japanese Archipelago and as a result, mere than a thousand persons suffered. The outbreak was Initially thought to result from polychlorinated biphenyl (PCB) poisoning1-* caused by the consumption of rice oil contaminated with KanocMor 400, a heat trentfar medium.' Subsequently, the "toxic" rice oil weseiso found to contain eievared levels of polychlorodibeniofurans (PCDFsJ*; highly toxic comaminants of PCBs*; and polyehlorinaud qutterphtnyH (PCQs)* -lexicologic ally ID-defined, but substances perhaps equally toxic as PCBs.1, * loth PCOFs and PCQs can bo financed when PCBs are hoattd.* The ratio of PCDFt or PCQs w PCBs can become high during the deodorlxaiion process of rice oil under high temperature and reduced pressure,1* Toxicolog- Ne*itnkr/Owtinkr Iftl lVet It (Ne. ()J HONS 2164*6 IcaJ contribution of PCDFs In Yusho wu further Implicated by discovering that the liver of Yusho patients who died from other ouiet after the outbreak of the poisoning con- pined PCDFs one Aounnd times higher than that found In normal populationi.**-11 A recant analytic of the blood of Yusho patients conducted 11 yrafter the Yuiho outbreak, detected PCQs but not PCD Ft, suggesting the role of PCQs' u a t uiable marker for the pact episode of exposure to toxic ofls.1* . Ourinp the iprin| and cummer of 1979, earn of peculiar ikln dilute chanctartaed mainly by acne; chsete-Mce die* charp from the Meibomian (land; pipnenation of the skin, plnfin and the nail bade; and abdominal paint wen report, td In two prefecture! in the middle part of Taiwan, Repub- Hc of China.11 Samples of tutpaetad "toxic" rice oil deli* end to ut In the autumn were found to contain PCBs. Gu chremato|raphic penarm of time tamploe were similar to those found In Japanese "toxic" rice oils. All Mood cam- plec from the patientc llkewice contilnod PCBs ranging from 54 to 1 36 ppb (unpubilched data). Tho number of patlantt exceeded 1,100 at the end of 1990; a detailed epidemiolog ical and clinical report will be made eltowhen. Of the 13 patientc In the present study, PCDFs wore found In the blood of aK except for on# write had mild clin ical manifestations. In Alt ctudy strong evidence au|foidn| a <ns)or role of PCDFs In developing clinical iranHimtiom and propodc which charectorite Yusho (not mery "pure PCB" poisoning) Is prownurf METHODS AND MATERIALS Throe samples of "toxic" oil, and lO-mi Mood camptas own coilecud from 1S patients wlthvsrlou dlnkai coven Itiai, who wore pupils Bring In the domsitorits on the school _ campus. Every mul was provided by tho school. The pupils were estimated to have consumed the contaminated oN In the school chow served during a 4-month period, up to 9 . months prior to the time of Mood sampling. Analytical methods for PCBs, PCQs, end PCDFs. Ten mllMiten of Mood were saponified wl A 20 ml of 2 N KOH ethanol solution for 1 hr under reflualng After sddldort of 30 ml wtter, the saponified sample wo* extracted with 30 ml of n-hexane. Then-hexane layer was than washed with 50 ml of water, dried over anhydrous lodhwa suifata and concantnted to ipproxlmateiy J ml In a (CD. concentrator. The n-hexane extract was put on s Florisll column (301 of 60-100 mesh Floruit, tctfvatlon at 1JO*C OvenWjht; 1.1 cm ID) snd eluted successhmly wltt'130 ml of n-hexane, 50 ml of 29 diethyl ether bt -hexane, 100 ml of 59 dkAyi ether In n-hexane, 330 mi of 509 dhthyl ether In n-hexane, and 250 ml of acetone at a rate-of appreximataly 2 ml/mln. The first and second eluates wore combined, concentrated, and analyted for FCBs by ps chromatography (GC| on a Varlan Aerograph 2100 machine equipped with SNI-ECD, as described previously.1 Polychlorinated quaterphsnyli eluted In the Aird frac tion were chlorinated exhaustively with antimony-penuchloride.xluned up on an alumina column to remora impurhirs, and determined by a SMmadsu GC-6A gu chromatograph fitted with "NI-ECO. Polychlorinated dlbcnipfurim were eluted In the fourth and fifth fractions from a Florist! column, and afur carafe! evaporation to drynoea were subjected to a final clean-up on an alumina column (301 alumina, Merck Co, Ltd, Art. He. 1077; 1J cm 10). To achieve more efficient removal of Impurities, 300 ml of 39 methylene chloride in n-hexane was used as the first afuaie. Polychlorinated dlboraoferans were recovered with 300 ml of 309 methylene chloride In n-hexane. The ektue was eveporated to dryneu and the residue was dissolved In n-hexane and analysed by GC using a Varian Aerograph 3100 gu chromatograph rnwhlne wlrh **NI-ECD. Gu chromatogrepb conditions wore u follows: column, 3 m X 3 mm glass column paused with 39 OV-17 on Gas Ovom Q (100/130 mash); infection. detaction, and column temperaturas, 253,330, and 250*C, respectively; carrier pa, Na (30 ml/min). hoiycMaehtsted dlbenaofurans were estimated by compering the peak height! of the gas chromatogram widt thoee of fiCDF standard, assuming that PCDF isomers wlA the tarns number of chlorine atoms all had the tame sensitivity, ragardtass of the sites of chlorine substitution (Fig 1). Polychlorinated Abontoferam In the samples went iden tified (Fig 3) by gaschromotomasspectrognphic (GC-MS) methods, using a JEOL-20KP gas chrematograph-)M$-300 man spectrometer wriA a computer system. The eonditiant wen as follows: column 3 m X 3 mm glas column pocked whh 39 OV-17 on Gas Chrom Q (MV100 mesh); Mot and column temperatures, 300 and 300*0, respectivoly; lontting energy, 70V; acceleration, 3.0 KV, carrier ps. Me (24) kg /cm*). GC-MS Oiromatopamsat m/a (M*)and (M+-C3) of PCDF in "toxic" oft A Taiwan (Fig 2). The PCDF frac tion of the oM wu Injected Into a GC-MS using a JIOL- 20KF gas chromuopaph-) MS-300 mass spectrometer with a computer system. The GC-MS conditions were u follows column, 2 m X 3 mm Men column pecked wlA 39 OV.l 7 on Gas Otrom Q (BO/100 mesh); Inlet and column temper, aturec, 300 and 300*C, respectivoly; lonlxing energy, 70 eV; acceloration, 3.0 KV; ourier gw, He (24) kg/cm*). ' RESULTS/ The gu chromatopaphk patterns of PCDFt (Fig 1) extracted and purified from toxic oil samples A both coun tries show fair resemblance In peeks 10 through 16, MflcatAg similarity of constituent homers between the two. A GC-MS confirmation ww also done oiTeach PCDF poak for toxic oil and Taiwamm blood samples. A similar relation ship was likewise present between the blood of a Yusho patient (No, 11) In Tehran and the Hvor of a Japanete patient. A GC-MS chromatographic analysis of the toxic ell from Taiwan indicates that peaks 3 through 14 reprtient 4-6 chlorinated dlbenzoferant with molecular Ion (M*) and fragmental Ion (M* - 63) resulting from the deletion of COO (Fig 2).* Tho Mood concentrations of PCBs, PCQs, and PCDFt found in pstienu from Taiwan and 7pan. Japanete worktrs occupationally exposed to PCBs, anil control Japanese sub jects are shewn in Table 1. PolycMorJntted dibenaoferans were detected in all tho Taiwanese patients (6 months after termination of toxic oil Inerstioni. except for one who hid 331 Motives ifEniiinrenlll Htiiih HONS 216**7 I I Fit. ]. Gm ittMUMpin af fCDf fntflaa: (A) Naak af Talwaaaaa *Ynto" aallaM (Na. 11); () B*ar af ) M-- "Y>>" aaiitar. (O "iaat" ail la Talvtn; (D) "laNt ail la Jiaan. CminwM af tack paak ikava la ia tkaaaNitpnit via caafbma* u fttiam ay OCMt analpiN.' paak Na. 2, lavarMaraMpiwftyl: Mk Nat. J-7, nmcMaratfkaMafank; paak Nat. I, f, 11, iM 11, paatatkiafaOaatafarta; aaak Nat. II, 14, tM 11, MataMaratfkamafaraa. Naitiakar/Datamaar 1*11 [Vat 1< (Ha. <)t MONS 216448 n) m/i(M*-0) m/i {241) m/t (275) m/a (309) ic m/i{M+) 40-CDF m/I {204) SO-CDP m/i (MS) 6O-C0F m/I (172) t (min) n*i(Ipw mw in Jw puoef Figaro 1. at a/l (II*) end (M*-n)errcDPIa<*Mea*aOlnTil*en. WdM|W the mildest cflnkal manifestation* and companding law concantratlona obaarvod among Tafwanare potfana In the kvois of PCOi and PCQt. cunom study (Tbbk 1). Whan occupational exposure to PCBt was terminated, their darmal laslont quickly improved'* In contrast to persistant efinkai signs retd symp DISCUSSION Although attampu to Identify PCDFs in die bleed of Jtpanna Yusho paliana 11 yr afur tha outbreak Mod, PCDF concantradoni exceeding there found In tha preaant toms among Yusho pedants that haw fingered nwre than 10 yr.M it should be again emphasised that most Japanese I Yusho patients now haw Wood PCS levoh that barely exceed there of the general popufeden.11 | caret, in relation to PCS Iavail, would haw baan found shortly after tha outbreak. Japanaaa teak oHa haw a graatar PCDP; PCS ratio, and roughly 10 drew Wgire eontrenlnitlon with both tubsancet than Talwanne toxk aiit (Tcbk 1). However, a direct aompaibon beteeen tha two out- bracks with retard to tha btttka of took wbnancat and kverity of poitoning is litiprealbll bacauaa only tome "toxic"' lot tamplas from tha Tafwanare Inddant are *v*S- (2) PCDFs show saong hepatotropUm," at do their analefua1' -potychlesinared dlbentodfexinet, White the PCDF to PCI ratio In tilt Wood ofTefwanew patients is approximately 1 ; 500, which Is similar to that found in Taiwanese toxic oils {Le, 1 : 300), the redo found In the liver of JcpanoM pallano, who succumbed 1 to 9 yr aftar the Incident, Is 1 : 5--a value far abow tha retire notad In the Japanese toxic oils (l: 100 In avenge (TaWe l)j. din- abia for analysis. > . What do the PCDF laval* noted In our study sljiifyf W believe that tha following evidence nrongty tuggaata that I PCOFiwve mainly retponaibk In tha paihoganaelt of I Yvtho. ' (1) There Individuals occupationally arpotad to pure Jcally, no abdominal pains nor Jaundice haw baan described among tha three tarlei of workers occupationally exposed to FCH**"1* while <5 to 73% of Japanaaa Yusho pationta experienced abdominal pains* and 11 % had jiundka accompanied b> edre abdominal tymptomt." Presenca of hepatorooic a**; htpatotoxic* PCDPi In Yusho can PCBt are usually tymptom-fraa; somaUmat mild dermal man* ifciuuont such at chJoracn* accompanied by mild contdtutienil tympiomt occur.**"1* Thb occurs despite that blood concentrations of PCgi In Individuals 7 yr aftar Hav ing thair workplace are *4uai to or yaattr by one digit than account for this ivtnptomatk dbcnpancy. (3) Although toxicity of PCDFs rarki dtpanrfing on tha biological panmatan (e.g, chkk embryo,* liver netrotii,a thymic atrophy,***** animal spociat,:i ate.) and on tha structural difference of isomers,*'***** most of there 174 ArxMoei ef CnWenimnUl Hunt MONS 216449  T ** TMp*MbTrtM 1* M T* M --*"***- 1 id m M u 0 M 0 M M M 4 M M Mi u n f? t M 11 UH4 WiW MMllKJ uu 1 M nu au MIIM lUtiia 4UN(H4i *CiPMri lMt MJ Ml X* 1M u tti ha 0 M IM U HJ IU Mi MJllUl Hill * M*4 ttTi fU IM MfMI ium^um fWlMi imi un ' W* ' MM Ml MM M fc *. MM UN MT MSI UM UM IMtWt Ml -- m -- U u IUiIM ut*n MillliMi Wi |1M Mi I M l Ml MiiTtrMI Mlllltill Miiti iid MiiM Hit WlMKlII Mil It i M4 MilMlMI MilMllM MitMiiM Mil it till MiiMtiM MilMlMM MilMHUI Mlflii* Mil<li MitMlM MilMll Mill! Mil Mfi -*Mt mii^iui 1 \ Itll (VvL. ]( (K#, )| HONS 21*450 3; loxiaoloptel comparisons were made uilng tht FCDF and which ranted "Kencml Yinhe". / Hrt Sec /race 1, )(u PC* umplrt which directly arrivoti "fresh" and "pure" from manufactures, UtI(Uln| PCOFt and PCIt that were either reproduced under the original confflOom (I.A, temper ature and coexisting air) at the rice oil production plant, or 7. Metf, la Ohanc, H.: KaaMmete, T.$ and FukuCk, Y. 1P7V. Mn tributlen and klelaglcal effect ef PCI, PCT end *CQ ee ne free /p Cenprrg Feed (In Japanmel Netcmbtr, If7*i If. a. Hart, L 1*10. atelepcel effete ef FClt end retted petycMer- purified from the "toxic oif-the causative agent of JapaneseYuiho-weobitrved that the toxicity of KDFtwif a hundredi to ten thousand times pester than PCS toxicity In monkey,* mouse* and ret.7 In addition to hepatooopinn of PCOFt, the blood PCDF: PCS ratio In the currant bated eewpeendi en awe monkeys. free ft* Jrmn. AAtrm im /cp H fpenrael. f J ** f. bHytU, H.I Msrrekawl, Yaend KeeMmese, T, 1*7*. OearrmUa- Uee pf pefychlertnatad euwpneyl (*CO) In Kassarai rtra ed that taraad "Ypera- and Inveetlguipn en PCQ ferasaden. / Feed HjrfSm /npae 111 417-34. ' study aieni one to the toxic contribution of PCOFt in 10. Mlytta, K, and ttaehlniese, T. 1P7>. Inraptiuslen en ereane- Yusho (4) Although the toxlcolo|lc role of PCQi has been ehSerimirC cewpeende farmed In ranaral Hat all the* raurad 'Ymhe". / Arad ffrp See To* 20i 1-#. 11. Mtyaa, H.; KarMmeae, Ta and Kanta, K. 1 *77. Oetoctiea and barely defined In Yusho, the present study thews that the derarmlnauenerpelyraueradieii--fenra tnenrmal human da- toxicity of PCQi It almost equal to that of PCIt.*'* Thorn- nrra and Kaneral Yiehe. / Feed Mn Sec yap II: 3CO-4S. fort, the blood levels of PCQi observed amon| Taiwanese Yusho patients tunest that these substances were relatively IX rachlmeie, Ta Hlysra, Ka and Kwdm,X. The prasanet ef palyMarinated eratarpnenyia In the tiewiwef Yaehe vldetae. Feed fifin/ {ifk pfw^* insignificant In ioxlcoio|icai contribution. Perhaps PCOs IX Chan, F.Ha Caw. PAa Wnn*. CKa and Own, C.|. IttC. Lerato A are more important In tniblinf one to differentiate whether and garehramttepipMc pettene ef peJvtMerlnated Wphenyla the patient hu had a put exposure to "Yusho-type" chlorHinaud hydrocarbons (I.a, PCSt, PCQi, and PCOFt) or to Monty PCSs.1' I in the Wned ef penane ef pwyehierinatnd Mphenm In ifw Mned e# petienw after fC* pelaenlfli In Taiwan fed (mtmt Facarraw Tuili/ 2J| J9|e3i. 1A Htrayawe. K. 1P7X AMedy ef PCI annssminetina ament InCuw trill werkere. Kummt Med Awec (w Jetenara) 41 s 1 -1A Submitted ter pukltratien Auiuet d, 1C1( accepted for pttMI- tie* Sepumecr ll,in. a lecurau rer rtprfnu metric be sent ter 6a OM, tCO,, FXD., Department ef Hypw* and Piimiln Madkiaa, toed ef Madklnt, Unfraiehy ef Tekye, 7-Xl, Hen|e. luntyo ka, Tokve 111, Japan. IX Here, la Hlrew. Ma Wteenebi. la YiktnMP, Ta Tebeiraihl, M.; and HldsHtnl, K, 1*7*. Heatth twee ef dm werban eraeprilen aly expend ta FC( at a pnlnt mensdacowini fanery. Amt Arp Orahc atrewnal frar Fw Weedh fin Japan--i) 17i 17-31, IX Osrw.KICa ikneeen, P.IL*and ItyeM, OX l7X Uuand baaM rffeeeef AreMer 13>l,a eih'ralihnewd blphaeyl. In mi elratrial rndiuy. Arab Xwdmw Hmm Mi 1**-OA 17. Hbtevenw. la WeradU. Yaend Kurenne, M. 1*77. Deterratne. den ef pehraMerfnated dtberwefxreiw In abmei ef peiiene with "Yuehn**. Fend Ceeraet TtiMkef 1 St IPX^X tmaiNcu. IX Fbraanne, Da f*, DJa drat. )a and m^nenhans, XIU 1*71. MeribMinn ef cbkb adene fbctdn kt Mak tbawA / AOA C S4t 12**-*X 1. Tsvktmete. MtkWemi, I.; Wrest, Ka KoPme,Ta Pekumaie, H-i Fvkeewie, Ka Kirntrara, **a wmirami, MatMbea, Ma Kt|sl, |a rae, Ya tewade, Ka Yeshiwere, Ha Ttonml, Ka Otvrl, KaShlwewe.Ha Dane, Ka KetoyetM, KiYene.T,; lie, . aaOkeU. Ta <a Ke|, TaTewlo. YaXefe.Ta Yew . Iti, Va Mly>e*d*l, M.}Seeae, M.; Htrf, Ka Tshtchlat. KjMew- eke. Kahskimwi, Y,;and SMtemeri,N. IMXTha ehtmiml luclet ee eatacUea ef teMt camaeuadl ks the rlw hn* elU usei by Uw pttlanu ef Yrabe.Fr*eA4cl McMra (la Jepen- *m) iOi m-fll X Tanabt, H. 1170. Ad has ftirdy an preraadea, dbfaneO and L-titmeat ef Yyihe, Study depart frara )epeawt A|eery ef Tkchnetefy O70|, I* (ijiMH, ]. KtHktivn*. M.; Yemirntw, Tj Meieeteta, la eedYeweitahl, A. ter:. taHrmteietitai u--y ee Yrahn.e pahonkte eetwed *y Inftnien l r l* eU cemewtetsed wtm a tewWeretU bned of eeiytniermetid eiphanyte. Xndran fbdhX Atmttt 1; 11*-21. 4. Kipvimt, Mtwde, Ya Md Kpreietme. M. 1*71. Chlprleeted iernteftitans In KaeeeMpr end flee ed seed by ppUemt whh "yv*he". /vave*4cie MedhP Mi SI>-*, i. Vet, ).Ca Kerman, H.H.; Viedef Meat, H.La sen Natter da raw. M.Ca and da Set, X. H. 1*70. idemlflcMlee end le-iae- l*|lckJ rnieaiien rf chiefinased dibaniefersn and cnleitneted naatiihtun* In saw aemiwalal pelyrhlertnaMd eiphenyh. Fend Cesmrr Tether : S2I-IX A suytll.M.; itaiMmeie.Ta and Kwnlta. N. 1P7I. DtieciSeeef unkne-n erlinetMeriniied aempeunda I* Kantwl llee ells 1*. AaM, ta Ked*. H.; Urabn, Ha rnd Trahent, X 1*7*. Darmesra leplraf lymptene ef Ysebe tHaraUenr In ebb derade. Faneehc Ace* Medic* (W Heeneei) 70t 173-OX 3X YecMmure, Ta Marl, Ha and Kentrant, IX1P7X hrai rwpcnra rctatlencWt between earde no latafec end aympterac in YraM patients: Shert rapen./ap y Hyp (In Jtptnrae) IX 111. ` 31. Kereten. Ma YecMnwn, Ta Mewnaefei. |.i end YcmapyoM, A. 1 *71, YMhn, a pntsnMnq cacned by Hee ell enmaralnetad wfth pelycMeHlwted MpbeeyX HSMMA Hta! At* ait 10M-01. 3X Vac, ).C- end rciwee. XK 1*70. Cerapemhe reMceleifcal ttedy with pMyrtlertensed Mphairyb IndMckera with ipecW lefenim te perphyrte.edeme fcrmetlen. Peer nirredi and Ua- rae rnldera. Tncfref Anpf Fkcrmcral 17t Ut-U. 3X OMaM, Sa Merita. Ma and fMirada, H. 1V7X Cerapantlm tenlclry ef pelyrhlerlnetad Mphcnyb end dtbaneefecara In rata. Tnclcar ApM Fherrancaf 4): 11-72. 2A Meart, |aa.; Cepu, XN.; end Vet, |.X 1tM. TtxMlty ef 3, S, 7,1 settschliredlbaapefraio-ptyllatiaarT rrraiu. free dctlenal Card FCI ffFA/ 77-OX 21. Meere. |.A; MeCenell, LX; DalprX O.W.; end Herrtc. tXW. 1 *7*. Cempemlm mxidry ef theb halepentlrd dibintefwme In tnlnta etft, etlca end rhraut wenkeyA AnneO HY Acad StL W. ). Mlclralten and A A. Meets, adu, pp- lil-IU. 3X Saakl, XiOuwe. Na end YraMmen. H. 1*77. Synthmle ef 3chlere and 1,4, karlcMrawdlbaniefiiraa and tnwr afreet en m< arewth ef mice and en the Hrar mlcrraemel drat metabeilalni ruymr tyucxe el raw. FadireW Asia afcdlec (In lapamM) ta: *-! OX iit Archlx; ef Cnvirenmental Haallti 216451