Yadda methylene blue ke shiga ƙwayoyin halitta: karɓar electron, sake rasa electron, da rarrabuwa a membrane

Idan ka kalli ƙwayoyin halittar da ke shimfiɗa cikin jijiyoyin jini suna shan methylene blue a kwanon gwaji, fassarar da ta fi fitowa fili ita ce kuma ba daidai ba. Ruwan renon yana rasa shuɗin launinsa, ƙwayoyin kuma suna samun ƙananan dunƙulen shuɗi; don haka za a ɗauka cewa rinin ya ketare membrane a matsayin shuɗin kwayar sinadarin. Amma gwaje-gwajen sun nuna akasin haka. Sigar shuɗin da ƙyar take ketarewa. Abin da ke ketarewa shi ne sigar marar launi, kuma shuɗin da ake gani a ciki yana samuwa ne a ciki, bayan isowa.

Wannan akasin abin da ake tsammani yana da muhimmanci domin yana ƙayyade abin da rini zai iya bayyana da abin da ba zai iya bayyana ba. Ƙananan dunƙulen shuɗi suna nuna rinin da ya sake rasa electron kuma aka tsare a ƙananan gabobin ƙwayar halitta. Ba sa nuna jimillar rinin da ke cikin ƙwayar, ba sa nuna adadin sigar da ta karɓi electron, kuma ba sa auna tasirin ilimin halittu. Don fahimtar dalili, bi matakai uku: karɓar electron a saman ƙwayar, yaɗuwar sigar da ta karɓi electron, sannan sake rasa electron da tsarewa a ciki.

Sigar biyu suna nuna halaye kamar sinadarai daban-daban

Methylene blue da ya rasa electron yana da tsarin zobba mai lebur da cajin lantarki tabbatacce. Yana narkewa cikin ruwa, yana shan haske sosai a kusan 600 zuwa 700 nanometers, kuma yana bayyana da launin shuɗi. Cajin shi ne muhimmin abu. Kwayar sinadari mai caji da kusanci da ruwa ba ta ketare membrane na lipid cikin sauƙi, don haka kalmar da ake amfani da ita cikin taka-tsantsan a wallafe-wallafen kimiyya ita ce ba ya ketare membrane sosai, ba wai ba ya ketarewa kwata-kwata ba.

Methylene blue da ya karɓi electron, wanda ake kira leucomethylene blue, ba shi da launi ko caji, kuma yana narkewa cikin kitse. Idan an cire cajin, membrane ya daina zama kamar katanga. Kwayar sinadarin da ta karɓi electron tana yaɗuwa ta ketare cikin sauƙi, kuma da zarar ta shiga wani yanki da ya fi sa sinadarai rasa electron, za ta iya sake rasa electron ta koma sigar shuɗi mai caji da ake tsarewa. Saboda haka, sinadaran redox na rinin su ne kuma tsarin safararsa. Canza yawan electron, kuma ka canza sauƙin ketarewar membrane.

Electron na matakin farko suna fitowa ne daga ƙwayar halittar kanta. Ƙwayoyin halittar dabbobi masu shayarwa suna safarar electron ta membrane na ƙwayar halitta, suna tura ƙarfin bayar da electron daga masu bayarwa a ciki zuwa masu karɓa a saman ƙwayar. Wannan aiki na safara ta membrane aiki ne na al'ada, wanda ake samu a yawancin nau'ikan ƙwayoyin halitta. Enzyme reductase da ke saman ƙwayoyin da ke shimfiɗa cikin jijiyoyin jini, wanda ke sarrafa rinin thiazine, yana da hali iri ɗaya da makamantansa a wani muhimmin fanni: yana ci gaba da aiki a gaban cyanide da azide a adadin da ke dakatar da matakan da ke biyo baya. Wannan bambancin ne ginshiƙin bayanin gaba ɗaya.

Rarrabuwa ta zo da farko, kuma ta kusan bayyana shigar rinin

A 1929, Marian Irwin a Rockefeller Institute ta yi wata tambaya mafi sauƙi. Wataƙila shigar rini rarrabuwa ce kawai tsakanin ruwa da shimfiɗun kitsen ƙwayar halitta, kuma ana iya amfani da shimfiɗar chloroform a madadin waɗannan shimfiɗun kitse a gwajin ɗakin bincike. A bincikenta na shigar rini ta hanyar auna shan haske, ta girgiza rinin da aka narkar cikin ruwa tare da chloroform, sannan ta kwatanta abin da ya koma cikin ɓangaren sinadarin organic da abin da ya shiga ƙwayoyin algae masu rai.

Sakamako biyu daga wannan aikin har yanzu suna da muhimmanci. Na farko, ruwan methylene blue na kasuwa yana ɗauke da trimethyl thionine, wanda yanzu ake kira azure B, kuma wannan gurɓataccen sinadari yana shiga da sauri fiye da methylene blue kansa, musamman a pH na alkali inda sigar free base mai narkewa cikin lipid ta fi yawa. Wasu abubuwan da suka yi kama da shigar methylene blue, a zahiri, shigar azure B ce ta zaɓi. Na biyu, pH yana ƙayyade sakamakon ta hanyar ƙayyade yanayin ionization, kuma ƙwayoyin da suka ji rauni suna barin rini ya shiga da yawa fiye da ƙwayoyin da ba su lalace ba. Ma'aunan rarrabuwa da yanayin caji ne suka sarrafa abin da tsarin gwajin wucin gadi da ƙwayar halitta mai rai suka sha. Kamanceceniyar da ke tsakaninsu ta goyi bayan ra'ayin cewa akwai shimfiɗu marasa ruwa a iyakar ƙwayar halitta.

Wannan samfurin ya yi hangen nesa game da lipid da ionization, amma bai iya bambance rinin da ke rarrabuwa a sigarsa ta asali da rinin da ke sauya ta hanyar sinadari a saman ƙwayar, ya ketare a wata siga, sannan ya koma sigarsa ta asali a ciki ba. Wannan bambanci ya buƙaci wani gwaji daban, wanda ya zo bayan shekaru saba'in a ƙwayoyin da ke shimfiɗa cikin jijiyoyin huhun.

Karɓar electron a waje, tsarewa a ciki

Merker da abokan aikinsa sun yi aiki da ƙwayoyin da ke shimfiɗa cikin jijiyoyin da ke kai jini zuwa huhun shanu, waɗanda aka rena a ɗakin gwaje-gwaje. Sun gabatar da jerin matakai uku: rinin da ya rasa electron yana karɓar electron a saman ƙwayar halitta ya koma sigarsa marar launi mai kusanci da kitse; sigar da ta karɓi electron tana yaɗuwa zuwa ciki; sannan sake rasa electron a cikin ƙwayar yana dawo da sigar da ba ta ketare membrane cikin sauƙi, wadda take taruwa. A bincikensu na shigar thiazine cikin ƙwayoyin shimfiɗar jijiyoyi, sun gwada wannan jerin matakai ta hanyar ƙoƙarin katse shi a takamaiman wurare.

Kayan gwaji na farko shi ne ferricyanide, mai karɓar electron wanda ba zai iya shiga ƙwayar halitta ba. Lokacin da enzymes na saman ƙwayar suka sa shuɗin rini ya karɓi electron ya koma marar launi a wajen ƙwayar, ferricyanide zai iya karɓe waɗannan electron daga gare shi, ya dawo da shuɗin rini a ruwan renon tun kafin ya ketare. Muddin ferricyanide yana nan fiye da rinin (10 micromolar na rini idan aka kwatanta da 50 zuwa 500 micromolar na ferricyanide), shigar rinin ta kusan tsayawa. Sai bayan ferricyanide ya ƙare ne shigar ta koma saurinta na al'ada. Saurin sauye-sauyen ya ba da wannan bayani ta wata hanya ta biyu. Ferricyanide yana raguwa da kusan tsayayyen sauri, wanda ya dace da ci gaba da karɓar electron a saman ƙwayar, tare da wani matsakaici da ake sake amfani da shi. A lokaci guda, rinin da ke wajen ƙwayar yana raguwa a tsarin exponential yayin da adadinsa yake ƙarewa. Tsayayyen gudana tare da adadin da ke ƙarewa yana nufin cewa alamar ferricyanide tana nuna ci gaba da karɓar electron, ba kwayoyin rini da ke fita daga ruwan ba.

Kayan gwaji na biyu ya kawar da matakin ketarewa gaba ɗaya. Ƙungiyar ta haɗa toluidine blue O, wanda yake kusa da methylene blue a rukunin thiazine, da doguwar sarƙar polyacrylamide mai girman da ba zai bar ta shiga ƙwayar halitta a kowace siga ta karɓar ko rasa electron ba. Duk da haka, ƙwayoyin sun sa wannan rinin da aka ɗaure ya karɓi electron. Saboda haka, karɓar electron ba ya buƙatar shiga cikin ƙwayar halitta. Na'urorin da ke gudanar da aikin suna a saman ƙwayar ko kuma suna da haɗin lantarki da shi.

Sai aka zo ga bambancin da ya ba takardar binciken sunanta. Cyanide da azide a 2 millimolar sun rage taruwar rini sosai, kuma a ƙarƙashin madubin ƙara girma, dunƙulen shuɗi mai duhu da na shunayya da suka saba kewaye cibiyar ƙwayar halitta sun kusan ɓacewa. Amma babu ɗaya daga cikin masu hana aikin biyun da ya rage saurin karɓar electron a wajen ƙwayar: zagayowar ferricyanide ta ci gaba ba tare da raguwa ba, kuma polymer da ba ya ketarewa ya ci gaba da karɓar electron. Toshewar tana a matakan da ke biyo baya. Cyanide da azide suna dakatar da sake rasa electron da tsarewar rini a cikin ƙwayar, ba karɓar electron a samanta ba. Gwaje-gwajen tantance ci gaba da rayuwar ƙwayoyin sun goyi bayan wannan fassara, domin ƙwayoyin sun riƙe enzyme lactate dehydrogenase, alamar da ke cikin cytosol, yayin waɗannan hanyoyin gwaji. Don haka, raguwar shigar rinin ba guba kawai ko fashewar membrane ce ta haifar da ita ba.

Wani gargaɗi daga takardar binciken har yanzu yana da amfani. Dunƙulen shuɗin suna kewaye cibiyar ƙwayar halitta amma ba a cikinta ba, wanda ya dace da rasa electron a cikin ƙananan gabobin ƙwayar. Amma binciken bai gano waɗanne gabobi ne ba. An kira irin waɗannan siffofi a wasu nau'ikan ƙwayoyin halitta na lysosome ne bisa hujja marar ƙarfi. A ɗauki tsarewar a matsayin abin da aka tabbatar, amma takamaiman wurinta a matsayin abin da bai tabbata ba tukuna. Masu karatu da ke bibiyar inda electron ke tafiya daga nan za su iya ci gaba zuwa safarar electron da sarrafa redox a mitochondria da kuma faɗaɗɗen bayani kan sha da rarrabuwar sinadari bisa hanyar shigarsa.

Rini ba ma'aunin adadin sinadarin da ya isa ba ne

Bincike na uku ya nuna abin da ke faruwa lokacin da aka haɗa wannan bayanin shigar rini da ma'aunin aikin ƙwayoyin halitta. Scott da abokan aikinsa sun gano cewa methyl beta cyclodextrin, sinadarin sukari mai siffar zobe, yana haɗuwa da methylene blue kai tsaye. Tsayayyen ma'aunin haɗuwar rukunin da ɗaya ya shiga cikin ɗayan yana kusan 310 per molar, idan aka kwatanta da kusan 5846 per molar na haɗuwar kwayoyin rini biyu. A ƙwayoyin fibroblast na L929, wannan haɗuwar ta hana ƙwayoyin ɗaukar rini da kuma ƙarfafa shan glucose: barin ƙwayoyin tare da cyclodextrin kafin ƙara rini ya hana shuɗin bayyana, kuma ƙara shi daga baya ya hanzarta gushewar rinin. Yayin da alamar shuɗin da ake gani da haske ke fita daga ƙwayoyin, safarar glucose da aka ƙarfafa tana raguwa a kusan lokaci da tsarin daidai.

Marubutan sun fassara wannan alaƙar a matsayin goyon baya ga hulɗa kai tsaye tsakanin rinin da wani furotin a hanyar kunna mai safarar glucose, maimakon wani tsarin damuwar oksideshon mai ɗorewa. Wannan fahimtar tana da ma'ana amma tana da iyaka. Gwajin bai bayyana sunan furotin ɗin ba, bai nuna haɗuwa da mai safarar kansa ba, kuma alaƙar da ke tsakanin gushewar rini da ɓacewar tasiri ba za ta iya kawar da duk wata hanyar da redox ke shiga tsakani ba.

Darasin da ya fi ɗorewa ya shafi aunawa. Rinin da ake gani ta haske yana nuna rini mai ganuwa wanda ya rasa electron kuma yake da alaƙa da ƙwayar halitta. Ba ya gano adadin sigar marar launi da ta karɓi electron ko kaɗan. Saboda haka, ƙwayar da launinta bai bayyana sosai ba har yanzu tana iya ɗauke da adadin rini, kuma ƙwayar da ta ɗauki rini sosai tana nuna rinin da ya rasa electron aka tsare, maimakon jimillar adadin da ya isa nama ko girman tasirinsa. Wannan taka-tsantsan tana aiki a mataki mafi girma ma: kwatanta rini tsakanin nau'ikan ƙwayoyin halitta, ko ɗaukar duhun yanki na nama a matsayin tabbacin cewa adadi mafi yawa ya isa wurin, yana maimaita kuskuren farko na kwanon gwajin. Shigar rini zagayowar karɓar electron, yaɗuwa, da sake rasa electron ce, kuma duk hoton sigar shuɗin a lokaci guda yana nuna ɓangare ɗaya kawai. Abubuwan da wannan ke nufi a aikace ga yin rini da sarrafa shi a ɗakin gwaje-gwaje da kuma fassara ikirarin adadin da aka bayar da adadin da ya isa nama suna fitowa kai tsaye daga wannan.