Iimvavanyo ezadibanisa i-methylene blue nokuphefumla kweeseli

Igazi lomntu elisuswe ifibrin, eligcinwe lifudumele ku-37 °C kangangeeyure ezintathu, lisebenzisa malunga neepesenti ezimbini zeoksijini eliyiquletheyo. Xa kongezwa isixa esincinane se-methylene blue ngaphambi kokuba ligcinwe kwezo meko, kwaloo gazi lisebenzisa malunga neepesenti ezingamashumi amathandathu anesihlanu. Akukho nto itshintshileyo kwiiseli. Yinto ekwisinyibilikisi kuphela etshintshileyo.

Elo nani livela kwiphepha likaGeorge Harrop no-E. S. Guzman Barron laseJohns Hopkins lango-1928, yaye lilapho yavela khona intetho ephindaphindwa ngokubanzi yokuba i-methylene blue iphucula ukuphefumla kweeseli. Le ntetho ayibobuxoki, kodwa intsingiselo yayo imxinwa kunokuba ivakala.

Oko inani elikwitheyibhile likulinganisayo ngokwenene

Into eyalinganiswa ekugqibeleni yayikukufunxwa kweoksijini, okwafunyaniswa ngokuhlalutya iigesi. UHarrop noBarron bahluza igazi elalisandul’ ukutsalwa ngoboya bomqhaphu ku-10 °C ukuze basuse iiseli ezimhlophe, bazalisa isampulu ngeoksijini, bayahlula, bongeza idayi kwinxalenye enye, baza bahlalutya zombini emva kokuzigcina kwiimeko zovavanyo. Amanani eeseli ezimhlophe ayengafane adlule malunga newaka kwi-cubic millimetre nganye. Ukusetyenziswa kweoksijini kwachazwa njengenxalenye yeoksijini yokuqala yesampulu eyayinyamalele.

Balinganisa nekharbon dayoksayidi, yaye kuloo nto indlela yokulinganisa yabazisela ubunzima. Igazi elisuswe ifibrin lalikhupha ukuya kwiimillimole ezimbini zekharbon dayoksayidi nge-100 mL nganye kwiyure yokuqala yokugcinwa kwiimeko zovavanyo, ngenxa yezizathu ezazinganxulumani nokusabela kwemichiza ababekufunda. Ngoko ke imilinganiselo yomlinganiselo wekharbon dayoksayidi ekhutshiweyo xa ithelekiswa neoksijini esetyenzisiweyo kwigazi elipheleleyo yayingasebenziseki. Kuphela ziiseli ezihlanjiweyo ezikwisinyibilikisi sikaLocke ezanika amanani anokutolikwa, yaye loo manani ayesondelelene phakathi ko-0.73 no-0.80, kukho idayi okanye ingekho.

Le nkcukacha ayinto engenamsebenzi. Kula maphepha, “ukuphefumla” kuthetha ukutshintshiselana ngeegesi ngaphakathi kwesitya esivaliweyo. Umsantsa phakathi kokusetyenziswa kweoksijini nokuphuma kwamandla asebenzisekayo uhlolwa ngokwahlukileyo kwinqaku elithi oko ukusetyenziswa kweoksijini kukuxelelayo noko kungakuxeleliyo nge-ATP.

Idayi yongeza kakhulu apho iiseli zaziphefumla kancinane kakhulu

Iiseli ezibomvu zegazi ezivuthiweyo zezilwanyana ezanyisayo phantse azisebenzisi oksijini inokulinganiswa. Iiseli ezibomvu zegazi zeentaka ezinenucleus ziyayisebenzisa. UHarrop noBarron bavavanya zombini, yaye lo mahluko usisiseko sengxoxo yabo.

Kwiisampulu zezilwanyana ezanyisayo idayi yandisa ukusetyenziswa ngokuphindwe phakathi kwamashumi amabini namashumi amahlanu. Kwigazi lerhanisi, elalineeseli ezazisele ziphefumla, ayizange iwandise umlinganiselo ngaphezu kokuphindwe kabini. Okukhona ukuphefumla kwasekuqaleni kwakuphezulu, kokukhona idayi yayinokongeza kancinane.

Kwalo mkhwa ubonakala nakuhlobo olunye lwesilwanyana. Isampulu yomntu onesifo se-polycythemia vera, eyayineeseli ezingekavuthwa, yasebenzisa i-11.3 yeepesenti zeoksijini yayo kungekho dayi, xa ithelekiswa ne-2.8 yeepesenti kwiiseli ezihlanjiweyo ezivela kumnikeli osempilweni. Idayi yayinyusa kuphela yaya kwi-63.4 yeepesenti, ukukhawuleza okuphindwe kathandathu apho iiseli eziqhelekileyo zakhawuleza phantse ngokuphindwe ngamashumi amathathu. Idayi yenza iiseli ezivuthiweyo zaziphatha ngokusondela kwiiseli ezingekavuthwa, nto leyo ababhali abathi yayiyenza.

Icyanide yayifanele ukukuthintela oko

Icyanide yayisaziwa ngokuthintela ioksideshini kwiiseli. UHarrop noBarron bayongeza nangona kunjalo, kunye nedayi.

Iziphumo zegazi lenkomo zingumzekelo wale meko. Isiqulatho seoksijini sehla ukusuka kwi-22.61 ukuya kwi-22.03 ngemithamo ngepesenti kwiiyure ezintathu kungongezwanga nto. Xa kukho i-methylene blue sehla saya kwi-12.84. Xa kukho i-methylene blue nepotassium cyanide ekuxinaneni kwemolar okuyinxalenye enye kumawaka amahlanu, sehla saya kwi-12.74. Umphumo wedayi phantse awuzange utshintshwe.

Umfanekiso: irekhodi eqhubekayo yokuphefumla kweeseli ezibomvu zegazi lomntu, elinganiswe kwinqanaba elincinane. Utolo luphawula ukongezwa kwe-methylene blue. Ngaphambi kwalo, umgca uphantse ube tyaba; emva kwalo unyuka ngokuqhubekayo. Iphinde yaveliswa kuHarrop noBarron, Journal of Experimental Medicine 48:207–223 (1928), PMID 19869478.

Isigqibo sabo yayikukuba idayi yayisebenza njengomthwali weoksijini, “ibonelela ngento eye yanyamalala kwiiseli ezibomvu zegazi ezingenanucleus zezilwanyana ezanyisayo ezindala.” Kwiminyaka emibini kamva, uMichaelis noSalomon baseRockefeller Institute bayichaza ngendlela eyahlala ihleli le mpikiswano. Umphumo wedayi wawunokuba “luphawu lwedayi oluzenzekelayo,” okanye idayi “yayingumzekelo wento ethile engaziwayo kwizinto eziphilayo.” Ezo ndlela zimbini zokuyitolika azikasonjululwa. Into etshintshileyo kukuba ngoku iiseli zinokuchazwa ngokuchane ngakumbi.

Isicatshulwa sesibindi senza kwaloo nto

UMichaelis noSalomon babonisa ukuba izinto ezitsalwe kumalungu omzimba ngamanzi abandayo zivelisa kwaloo mkhuthazo. Isibindi sempuku sasinamandla kakhulu, silandelwa zizintso, udakada, netyhalarha. Ubuchopho, izihlunu, nenxalenye yolwelo lwegazi zazingenampembelelo okanye zinempembelelo encinane.

Emva koko bavavanya eso sicatshulwa kukho ikharbon monoksayidi. Ukuphefumla okubangelwa sisicatshulwa okanye yidayi akuzange kutshintshe kumxube oneepesenti ezingama-98 zekharbon monoksayidi neepesenti ezi-2 zeoksijini. Kwaloo mxube weegesi wathintela ukuphefumla kwegwele nokweeseli ezibomvu zegazi zeenkukhu ezinenucleus malunga neepesenti ezingama-60 ukuya kuma-80, ngokuhambelana nenkcazelo kaWarburg ye-enzyme yokuphefumla ebuthathaka kwikharbon monoksayidi. Ngoko ke into engachongwanga ekwizicubu yayingeyiyo i-enzyme kaWarburg.

Izinto ezininzi ezazikrokrelwa zakhutshelwa ngaphandle njengoko uphando lwaluqhubeka. Iglutathione ekwimo encitshisiweyo necysteine zavelisa kuphela ukusetyenziswa kweoksijini okuyimfuneko kwioksideshini yazo. Icoenzyme kaHarden noYoung yakhutshelwa ngaphandle kuba izihlunu ziqulethe eyona ninzi yayo kodwa zabonisa phantse owona mphumo mncinane. Icytochrome kaKeilin, eyayithiywe kwiminyaka embalwa ngaphambili, yahlangabezana nokuphikisana okufanayo. Olwabo uluvo, lokuba into esebenzayo yayivela kwinucleus yeseli, balubiza ngokuba yingcebiso nje.

Umphumo wawuxhomekeke kwiimeko ezithile

Iiseli kwakufuneka zihlale ziphelele. Ukugqabhuka kweeseli ezibomvu zegazi ngamanzi adistiliweyo okanye ngokuzikhenkceza ngokuphindaphindiweyo kwaphelisa ukusabela kwazo kwidayi nakwisicatshulwa, yaye ukugqabhuka kwenxalenye yeeseli kwakunciphisa ngokomlinganiselo.

Neswekile yayiyimfuneko. Iglucose, ifructose, nesucrose zazixhasa ukuphefumla okubangelwa yidayi; imannitol yayingakwenzi oko. Xa kungekho swekile kwakungekho kuphefumla konke konke, yaye idayi yayibonakala izonakalisa iiseli ngoxa loo nto isenzeka, ngoko iglucose eyongezwa emva kweyure enye okanye ezimbini ayizange ikubuyise. Ababhali bacebisa ukuba idayi yayisenza ioksideshini yecandelo lesakhiwo elifunwa yiseli.

Ukukhanya kwawandisa kakhulu umphumo wedayi, nangona umphumo wawucacile nasebumnyameni. Ezinye iidayi ezinokuncitshiswa ngokwamkhemikhali zaziziphatha ngokwahlukileyo: itoluylene blue yalingana ne-methylene blue okanye yayidlula, ngoxa i-indigo disulfonate yenza phantse kungabikho nto. Akuzange kuvele unxulumano olucacileyo phakathi kwamandla edayi okwenza ioksideshini nokuncitshiswa nobukhulu bomphumo wayo; ababhali bathi oku kwakubangelwa kukukwazi kwayo ukudlula kwinwebu nobutyhefu bayo.

Umphumo walandela ukubila, hayi ukuphefumla

Uphando lukaBarron lwango-1930 lwasuka kwiiseli ezixhonywe kulwelo lwaya kwizilayi zezicubu zamalungu aqhelekileyo empuku nakumathumba afakelweyo.

Kwizicubu ezingenayo iglycolysis eyenzeka kukho ioksijini, i-methylene blue ayenzanga nto okanye yacinezela kancinane ukusetyenziswa kweoksijini: kwizintso malunga neepesenti ezi-14, kudakada lwesisu malunga neepesenti ezi-14, esibindini malunga neepesenti ezi-5. Kwizicubu ezibilayo kukho ioksijini, yandisa ukusetyenziswa: kudakada malunga neepesenti ezi-8, ebuchotsheni malunga neepesenti ezili-13. Ukwanda kumathumba kwasuka malunga neepesenti ezingama-27 kwi-carcinoma yebele lomntu ukuya ngaphezu kweepesenti ezingama-80 kwi-Rous sarcoma yenkukhu, ngokusondele kumlinganiselo weglycolysis eyenzeka kukho ioksijini kwizicubu nganye.

Umfanekiso: ichaphaza ngalinye luhlobo olunye lwezicubu. I-eksisi ethe tyaba yiglycolysis eyenzeka kukho ioksijini; i-eksisi ethe nkqo lutshintsho ekusetyenzisweni kweoksijini emva kokongezwa kwe-methylene blue. Idatha evela kuBarron, Journal of Experimental Medicine 52:447–456 (1930), Itheyibhile 6, PMID 19869777.

Uvavanyo lwecyanide lwavelisa umfanekiso ochaseneyo. Xa ukuphefumla kwezo zicubu zazingasabeli ngaphambili kwathintelwa ngecyanide kwaza ukubila kwabonakala, idayi yandisa kwakhona ukusetyenziswa kweoksijini: kwizintso malunga neepesenti ezingama-59, esibindini malunga neepesenti ezili-19, kudakada malunga neepesenti ezingama-286, kwityhalarha malunga neepesenti ezingama-200.

UBarron wacebisa ukuba idayi inokusetyenziswa njengovavanyo lwamandla okubila eseli okanye ezicubu, waza waqukumbela ngengqikelelo: izizukulwana zeeseli zethumba ezikhuliswa kwisondlo esinedayi zisenokuthi ngenye imini zenze imetabolism yethumba ifane ngokusisigxina naleyo yezicubu eziqhelekileyo. Lowo yayingumbuzo wophando owabhalwa ngo-1930. Asilobango lonyango.

Oko iimvavanyo ezazingenakukulinganisa

Ngo-1928 nango-1930, “ukuphefumla” kwakuthetha ukusetyenziswa kweoksijini. I-adenosine triphosphate yachazwa okokuqala ngo-1929, yaye ukuboniswa kokuba iimitochondria ezahluliweyo zenza iibhondi ze-ester ngephosphate ngoxa zidlulisela iielektroni kwenzeka ngo-1949. Akukho nalinye kumaphepha okuqala nge-methylene blue elalinokulinganisa ukuveliswa kwamandla, yaye akukho nalinye elathi liyakulinganisa.

Iiseli yingxaki yesibini. Iiseli ezibomvu zegazi ezivuthiweyo zezilwanyana ezanyisayo azinazo iimitochondria. Ngexesha lesigaba sereticulocyte zisusa iimitochondria neeribosome ngokwaphula nokuphinda zisebenzise ngokukhethayo izinto ezingaphakathi kuzo, nto leyo eyenza iseli ebomvu evuthiweyo isebenze ngeglycolysis ize ixhomekeke kwindlela yayo yepentose phosphate ikakhulu kwi-NADPH kunakwi-ATP. Kwiimvavanyo zango-1928 kwakungekho tyathanga lokuthutha iielektroni elalinokulungiswa. Nantoni na eyayisebenzisa ioksijini yayisenza enye into.

Ngoku loo nto ichazwa ngokucacileyo ngokwaneleyo. I-methylene blue yamkela iielektroni kwizinto zeseli ezibonelela ngamandla okunciphisa ngokwamkhemikhali ize ibe yi-leucomethylene blue, ethi iphinde yenziwe ioksideshini yioksijini yemolekyuli, ngaloo ndlela igqibe umjikelo. Kwiiseli ezibomvu zegazi lomntu, iflavin reductase exhomekeke kwi-NADPH needisulfide reductase ezinxulumene nayo zenza inyathelo lokunciphisa. Uphando lwango-1976 olusebenzisa iglucose ephawuliweyo lwafumanisa ukuba idayi ikhuthaza indlela yehexose monophosphate, ukuba umphumo kwinqanaba eliphantsi lokuxinana uxhomekeke kwi-oxyhemoglobin, nokuba ukuxinana okuphezulu, 0.1 mM, kuvelisa ihydrogen peroxide. Idayi encitshisiweyo edibana neoksijini ivelisa iperoxide, hayi iibhondi zephosphate.

Kwiimitochondria imeko iphikiswa ngakumbi kunokuba iingcaciso ezishwankathelweyo ezithandwayo zivuma. Olunye uthotho lophando luthi idayi ithatha iielektroni kummandla wecomplex I ize izinike icytochrome c. Olunye lwafumanisa ukuba kwiimitochondria zobuchopho bempuku ayinako ukubuyisela ukuphefumla emva kokuthintelwa yi-antimycin, lwaza endaweni yoko lwabeka umamkeli wee-elektroni kwindawo yokubophelela i-ubiquinol kwi-complex III. Olwesithathu lwafumanisa ukuba ukuxinana okuphantsi kwandisa ukusetyenziswa kweoksijini xa ziphumle kodwa akuzange kutshintshe ukuphefumla okukhuthazwa yi-ADP. Eyona nkcazelo ithe ngqo yale nto yeyokuba ukusetyenziswa kweoksijini okubangelwa yi-methylene blue akuxhomekeki konke konke kwifosforileshini edityaniswe neoksideshini: kwaqhubeka xa ityathanga lokuphefumla lithintelwe, nakwiisampulu ezingenalo.

Kukho nemilinganiselo ethe ngqo yomsantsa phakathi kwezi zixa zimbini. Kwiimitochondria zesibindi sempuku ezahluliweyo, ukwanda kokuxinana kwedayi kwehlisa imilinganiselo yolawulo lokuphefumla kwaza kwabangela ukudumba, nto leyo ababhali abayichaza njengokuqhawulwa konxibelelwano kwifosforileshini edityaniswe neoksideshini. Kwisibindi sempuku esasidluliselwa ulwelo, i-methylene blue yandisa ukusetyenziswa kweoksijini ngoxa umlinganiselo we-ATP xa ithelekiswa ne-ADP wawusihla.

Iingxelo zekliniki zinomyalezo ofanayo. Imethemoglobinemia inyangwa malunga nemiligram enye ukuya kwezimbini ze-methylene blue ngekilogram nganye, efakwa emthanjeni, ngokusebenzisa indlela yokunciphisa ngokwamkhemikhali exhomekeke kwi-NADPH, yaye iidosi ezinkulu zibangela kwaloo meko inyangwayo. Kwaloo ngxelo yohlalutyo ithi umphumo wayo njengento echasana nobutyhefu mkhulu kakhulu kwiiseli ezibomvu zegazi ezipheleleyo, uze unciphe xa kukho ukugqabhuka kweeseli zegazi. Eso sisiphumo sango-1930 esiphathelele ukugqabhuka kweeseli esiphinda sibonakale emva kweminyaka engamashumi asixhenxe anesithathu kwimeko yasesibhedlele. Ikhompawundi enemiphumo eluncedo neyingozi kumacala ahlukeneyo omda wedosi ayiyokhompawundi ekufanele kuqikelelwe idosi yayo kumlinganiselo wokusetyenziswa kweoksijini.

Isizathu sokuba ibhetshi ibandakanywe kwingxoxo

UHarrop noBarron baphawula into efanele ukufundwa okwesibini. I-methylene blue ngokwayo ivelisa ibhendi kwispectroscope phakathi kweebhendi ezimbini eziphawula imethemoglobin, “nto leyo enokubangela ukudideka ngokulula xa kuhlolwa nje ngokukhawuleza.” Idayi inokulinganisa into ekukhangelwa yona.

Kwale ngxaki inendlela eyenzeka ngayo ngokwamkhemikhali. Iphenothiazine enxulumene nayo, ekhoyo njengesithako esincinane, ineendlela zayo zeoksideshini nokunciphisa, ukukwazi kwayo ukudlula kwinwebu, nobutyhefu bayo. Omnye umzekelo obhaliweyo ubonisa ngokucacileyo ukubaluleka koku: kuphando lokufakwa kwe-iodine kwi-methylene blue, iisampulu ezazineepesenti ezimalunga ne-7 ukuya kwezi-8 ze-Azure B zavelisa ikakhulu i-Azure B efakwe i-iodine, ngoko yayibukungcola, hayi i-methylene blue, obabulawula ukusabela kwemichiza.

Nabani na osebenzisa i-methylene blue njengesixhobo sokusabela kwemichiza, elebhu, kwibhentshi yokudaya iisampulu, okanye kwitanki yeentlanzi, usebenza ngesithako esingachazwanga kakuhle ngaphandle kokuba ibhetshi iye yahlolwa yachazwa. Izinto ezinxulumene nayo, ukuqinisekiswa kokuba yintoni, ukungcola okuziielementi, izinyibilikisi ezishiyekileyo, nentsalela emva kokutshiswa ziyinxalenye yovavanyo kunokuba ngamaxwebhu nje alujikelezileyo. I-Blupreme ifumana i-methylene blue yayo kumenzi wamayeza ize ipapashe isiqinisekiso esipheleleyo sohlalutyo, ivavanya yonke imigangatho ye-USP kunesinyithi esinzima kuphela; isikhokelo sokufunda isiqinisekiso sohlalutyo kunye noko ukuchazwa njengomgangatho we-USP kukuqinisekisayo noko kungakuqinisekisiyo zichaza indlela yokuthelekisa uxwebhu nebango.

Inani lalichanile

Iimvavanyo zango-1928 nezango-1930 zenziwa ngononophelo, yaye eyona nto yaphawulwayo kuzo isaqinisekisiwe: idayi enokuncitshiswa ngokwamkhemikhali inokwenza iseli ephantse ingaphefumli isebenzise ioksijini ngokuphindwe kaninzi kumlinganiselo wayo xa iphumle, ngaphandle kokudlula kwinyathelo elithintelwa yicyanide.

Impazamo ivela kamva, xa “ukuphefumla” kufundwa ngentsingiselo yako yanamhlanje. Umlinganiselo wotshintshiselwano lweegesi ukuxelela ukuba iielektroni zifike kwioksijini. Awukuxeleli ukuba nayiphi na kuzo isetyenziselwe ukwenza i-ATP, yaye kwiseli engenazo iimitochondria umsantsa phakathi kwezo ntetho zimbini mkhulu kangangoko unokuba njalo. Imephu ebanzi yoko i-methylene blue ikwenzayo kwiiseli ibeka kwa-lo mahluko kumxholo obanzi, yaye imbali yale khompawundi ukusuka kwidayi ukuya kwiyeza ichaza indlela umbala wamalaphu owafikelela ngayo kwezi mvavanyo kwasekuqaleni.