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Distribution of Abnormal Hemoglobin around the Coast of the Seto Inland Sea in Comparison with Those of Surrounding Countries

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、 KawasakilkaishiArts&Sci(20):59-63(1994) DistributionofAbnormalHemoglobinaroundtheCoastoftheSeto InlandSeainComparisonwithThoseofSurroundingCountries ToshikoYAMASAKI,KazuoHIDAKAandlwaoIUCHI D"α”"29"ォqfBiOC〃e"ZfSォ7恥KtzwaSα〃ノ'ん碗azISじんo01 K2"rzsh洗j7DZ-OZ,ノZ功α泥 (ReceizノGao〃Sセカ彪刀2627・3QZ994ノ 5 9 Abstract About760sortsofabnormalhemoglobins(abnHbs)havebeenreportedinthe world(1994).Wecontinuedtoscreenthebloodspecimensofinhabitantsaroundthe SetolnlandSeaforthepastl5years.ThedetectionrateofabnHbofourlaboratory wasdifferentfromareatoareawithanaverageofoneperl,800whichareslightly higherthanoneper3,000to4,000inJapan.Morethan30differentabnHbsasthe summationofthereportsoffivelnstitutehavebeenregisteredintheseareas.However, neitherRacialabnHbslikeHbSnorspecificvariantscommontoalloverJapanwere detected. Introduction SincethefirstabnHb(HbM-Iwate)wasreported'),2)inl960,thescreeningofabn Hbhasbeenconducted3)'!')'5)onaboutamillionpeoplebythelaboratoriesorhospitals locatedatFukuoka,Ube,Hiroshima,Kurashiki,Matsuyama,Osaka,GifuandTokyo(in orderfromwesttoeast).DetectionofabnHbbyelectrophoreticscreeninghasbeen carriedoutactivelyespeciallyintheareaonthecoastoftheSetolnlandSea.Recently someabnHbshappenedtobedetectedbyHPLCforquantitationofHbA,c,however, theelectrophoreticscreeningprovidedamostefficientmethodofstudyingepidemiologic charactersofabnHbs.Sincel974,wehavecontinuedtoscreenthebloodspecimensof inhabitantsaroundtheSetoInlandSea.Thepresentpaperaimstodescribeabrief reviewofourresultofthescreeningsurveyonthebasesofthoseoftheaforedescribed institutes. MaterialsalldMethods Icecoldbloodspecimensusedfortheexaminationwereshippedtouseveryonce aweekfromKagawaPrefecturalCentralHospitalinKagawaprefecture,Fukuyama NationalHospitalinHiroshimaprefecture,KobeCentralHospitalinHyogoprefecture andKawasakiHospitalinOkayamaprefectureduringtheperiodfroml974tol989.All ofthemwerelocatedincloseproximityfromourlaboratory.Eachspecimenswere registeredintocomputerwithitemsoflivingplace,name,ageandsextoavoiddouble testingofthesameindividuals.VenousbloodwascollectedinHeparinsolution.The anticoagulatedbloodwasmixedwith0.3%KCNsolutionwhichwaspreviouslyputin

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Qbl 6り KawasakilkaishiArts&Sci(20)1994 ahollowofal50wellplateandthenthemixturewasplacedtofreezeat-70℃ overnight.Whenmeltingthemusingthesocalled"freezeandthawmethod'',150 hemolysateswerepreparedatonce.One"lofhemolysatewasappliedonacrylamide IEFplate(5%T,2.6%C,pH5-9,10×17cm)andlEFwerecarriedoutat4℃,500Vfor 2-3hr.About80hemolysateonanlEFplatewerepossibleapplication. Thepreparationofglobinandits"andnchain,enzymaticdigestionofthechain andaminoacidsequencingforprimarystructuralanalysiswereperformedbythemost conventionalwayofmethodologyatontime. Result Theresultofthesurveywhichwascarriedoutinourlaboratoryforthepast20 yearsisshownintheTable.InTakamatsuarea,73,536peoplehadbeenscreenedand51 TableTheIncidenceofAbnormalHbsFoundaroundtheSetolnlandSea

Takamatsu Fukuyama Kobe

1974.1‐1988.12 73,536 19854-1988.6 31,419 19817‐1983 47,458 15 1/3164 10 Ube-2 Nunobiki Albany-Suma Handa JHabana Umi N.1. 5 Syracuse GCoushatta Ankara Riyadh N.I No.ofExamined* No.ofabn.Hb Incidence 33 1/952 51 1/1442 "chain 16 Ube-2 Mizushi N.I. 17 Ube-2 St.Lukes N.I. 114 1 917

3111112

の芦画掴両崖 "chain 27 Takamatsu Yusa JBangkok GSzuhu N.I、 1 Fukuyama

93113

1 1

11111

ychain 0 5 FFukuyama F-ForestPark N.I. 0 1 1 3 N.I 8 10 0 N.I * notidentified Thenumberisrefertocomputerregisteredindividualswithitemsoflivingplace,name, ageandsextoavoidoverlappedexamination.

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= ToshikoYAMASAKIetal:ScreeningofAbnormalHemoglobin 6Z abnHbsweredetected・Detectionratewasthereforelover1,442whichissomewhat higherthananaverageinJapan(1/3,000-1/4,000).ItisnoteworthythatHbUbe-2("68 Asn→Asp)andHbTakamatsu(Pl20Lys→GIn)wassporadicallydistributedinslightly highfrequency,whichare"andPchainvariant,respectively.InFukuyamadistrict, 31,419peoplehadbeenscreenedand33abnHbsweredetected.Detectionratewasl/ 952whichissignificantlyhigherthantheaverageinJapan.HbUbe-2wasarelatively highfrequencyindicatingthereforethatismorewidelydistributivethanHbTakamat-su.InKobearea,47,458peoplehadbeenscreenedandtherewerel5abnHbs. Detectionratewasl/3,164.TherewasnotanycommonlyabundantabnHbsinview offrequency,butmanykindsofvariantsweredetected.InOkayamaarea,12,678 peoplehadbeenscreenedand4abnHbsweredetected(dataisnotshown).Detection ratewasl/3,170.ThethreeabnHbswereHbUbe-2,HbHoshida(P43Glu→Gln)and HbFukutomi(Q,126Asp→Val)OneremaimngabnHbwasnotidentifiedyetforthe sakeofaverylimitedamountofsample. Discussion AccordingtothelistofthelnternationalHemoglobinl㎡ormationCenter(IHIC)in 1994'),about760kindsofabnHbshavebeenregistered.Sincel948,thenumberof papersforthenewabnHbsandthescreeningsurveyhavebeenincreasedyeartoyear intheworldreachingaplateau. InChina,afairlylargenumberofpeople(About800,000peopleintotal)inthe provinceswerereportedtobescreened7)'8)'9)'lately.DetectionrateofabnHbineach provincerangedfromO.05tol.09%,whichwerehigherthanthatofJapan(0.03%).The kindsofabnHbwerereportedtoberatherfew,24intotal,inspiteofsuchhigh frequencyandlargescalescreenedpeople・HbEandHbNewYorkweredistributed

inprettyhighfrequency(HbEwas66%andHbNewYorkwaslO%oftotalabnHb)71

HbEisthemajorabnHbespeciallyinthesouthpartofChina. Korea,thenearestneighboringcountrytoJapan,hasrarescreemngreportsofabn Hb. InSoutheastAsiancountries,thefreguencywasexpressedonlyintermofHbE, andineachcountry,exceptforSingapore,thescreemngwasconductedonasmall numberofpeople'0),thereforeitisnotpropertocomparewiththedataofJapan.But itissurethatHbEexistsinveryhighfrequencyinthesecountries(0.2-34.9%).This racialabnHbisnotdistributeduniformlyineachcountry,thatis,thehighest occurrenceareseeninLaosandCambodia(34.9%inbothcountries). InJapan,aboutl60kindsofabnHbswerefound.Seventeenofthemwere seriouslyunstableHbvariantswhichcausehemolyticorcyanoticdisorderevenin

heterozygouscondition.About50ofthemdemonstratedabnormaloxygenaffinity.

Morethan60%ofthemweredetectedinpeoplearoundtheSetolnlandSea.Aboutl4

millionpeoplehavebeenestimatedtolivearoundtheSetolnlandSeaand460thousand

peopleintotalhadbeenscreenedbysixlaboratorieslocatedthisarea.Theyperformed quitealargescaleandexactsurveywhichwasneverreported,sinceabout3%ofthe totalpopulationwassUbjectedtothescreemngandprovidedamoreefficientand ー

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" 髭 KawasakilkaishiArts&Sci(20)1994 reliableepidemiologicalstudy.Twentyyearsago,westartedthescreeningtestwith useofthebloodspecimenofmhabitantsaroundtheSetolnlandSea(Table).Detection ratesofHbvariantsintheTakamatusandtheFukuyamaarea(1/1,442andl/952, respectively)werehigherthantheaverageofJapan(1/3,000-4,000).Therewerelocally specificandcommonabnHbsintheseareas,thatwasHbUbe-2andHbTakamatsu. Ontheotherhand,therewasnotsporadicabnHbintheKobeareainviewof frequencybutvarietiesofabnHbwereabundant.Thesethreeareasarelocatedonthe coastofthelnlandSea,thereforethepresenceofcommonvariantsinonelocationmay denotethattherehasbeenaveryminutegeneflowuptothelateerainTakamatsu andFukuyama,therefore,assumptionofthelocationoriginofabnHbmaypermitus

tospeculatethatitisthesamelocationaswhenweverifieditsendemicityevenifthey

areportto,〃ns.

InthecaseofHbTakamatsu,wedidfamilystudybycarefulinquiry.Theyhad

nobloodrelationshipinatleast4to5generations.Thepresumptionthatallcarriers

havedescendedfromasinglecommonancestryseemsaprobablespeculation.Itwas

alsonoteworthythatthisrarelyoccurringHbvariantspossessonehomozygouscarrier

withoutclinicalmanifestations.

Inconclusion,manykindsofrareabnHbswerereportedbut#(RacialabnHbs"like

HbSinNegroesandHbEinSoutheastAsianshavenotbeendetectedinJapan.

However,somevariants,forexampleHbK61n''),HbM-lwate,HbUbe,HbM-S

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tothinksomeothersareimmigratedanddwelledfromthemainlandofAsia.For

instance,inthecaseofHbHofu,itisarareHbvariantinJapan,butitisnotrarein

India,thatis,theabnHbmighthavecometoJapanbywayofbothseaandroadfrom

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existinanothercountryinAsia,too.Infact,HbEisthemainHbvariantinSoutheast

Asia'0)anditisoftenseenwithadeclivityinthesouthpartofChina,thenminimum

toJapan.ItmightbesuggestionthatabnHbcanbetransportedbyroadthroughthe

politicalboundary.

Infuture,itwillbeinterestingtoreinvestigatethedistributionofabnHbaround

thecoastoftheSetolnlandSeaafterdeterminationofthehaplotypeofcommonabn

Hbgene'2),'3)orothergeneticmarkerssuchasHLAandbloodgrouptoknowtheexact

originofourrace.

ThisstudywassupportedinPartbyaResearchProjectGranttol.I.(No.5-102and

6-103)fromKawasakiMedicalSchool. Reference 1)CarverMFHandKutlarA:InternationalHemoglobinlnformationCenterVariantList. Hemoglobinl8(2),1994 2)MiyajiT,Iuchil,ShibataS,TakedalandTamuraA:PossibleAminoAcidSubstitution intheafChain(a,87.ryr)ofHbMIwate.ActaHaematolJap26:538-543,1963

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5 3 ToshikoYAMASAKIetal:ScreeningofAbnonnalHemoglobin SaitoS:StudyofAbnormalHemoglobininEhime,Japan5TwoStructuralVariantsof HemoglobinA(HblandHbJlran)andStructuralandSyntheticVariantsofDeltaChain. JpnJHumanGenet29:335-351,1984 Sumidal:StudiesofAbnormalHemoglobinsinWesternJapan.FrequencyofVisible HemoglobinVariants,andChemicalCharacterizationofHemoglobinSawara(Q#"26A'aA)and HemoglobinMugino(HbLFerrara;"47G'yA).JapJHumanGenetl9(4):343-363,1975 OhbaYandMiyajiT:AbnormalHemoglobinsandThalassemiaS]mldromeinJapan. Byohtaiseiril(4):382-388,1982 HidakaK,IuchilandShimasakiS:StudiesontheabnormalHemoglobinⅨ二An ImprovedMethodofHighPerformanceLiquidChromatographyforSeparationand IdentificationofTrypticPeptidesofAbnormalHemoglobin.KawasakilkaishiArts&Sci lO:49-54,1984 TaoZY:HemoglobinopathiesinChinaMainland.Hemoglobin5(5):517-524,1981 JenPC:AbnormalHemoglobinsFoundinGuangdong,P.R.China.Hemoglobinll(6): 573-579,1987 LiHJ,LiuDXetal:HemoglobinopathiesinXinjiang.HemoglobinlO(4):435-449,1986 FucharoenSandWinichagoonP:HemoglobinopathiesinSoutheastAsia.Hemoglobinll (1):65-88,1987 1uchil,HidakaK,etal;IdentificationofabnormalHemoglobinK61n("98(FG-5)Val-> Met):TheFourthHbK61nVariantFoundinJapan.KawasakiMedJ5(1):61-69,1979 ShimizuK:DNAPolymorphismPatternsinthePGlobinGeneClusterintheJapanese Population・Hemoglobinll(5):487-496,1987 ShimizuK,ParkKSandOmotoK:TheDNAPolymorphismsoftheB-GlobinGeneCluster andtheArrangementsoftheQf-andthey-GlobinGenesinKoreans.Hemoglobinl3(2): 3 ) 4 ) 5 ) 6 ) 11 78 9 ) 10) 11) 12) 13) 137-146,1989 ﹄の 』

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