Kuyedza kwakabatanidza methylene blue nekufema kwemasero

Ropa remunhu rakabviswa fibrin, rikadziyiswa kusvika pa37 °C kwemaawa matatu, rinoshandisa zvikamu zviviri kubva muzana zveokisijeni yarinaro. Kana methylene blue shoma yawedzerwa risati rachengetwa pakudziya ikoko, ropa rimwe chete rinoshandisa zvikamu zvinenge makumi matanhatu nezvishanu kubva muzana. Hapana chakachinja pamasero. Musanganiswa wemvura chete ndiwo wakachinja.

Nhamba iyoyo inobva mubepa ra1928 rakanyorwa naGeorge Harrop naE. S. Guzman Barron kuJohns Hopkins, uye ndipo pakabva chirevo chinodzokororwa kakawanda chekuti methylene blue inovandudza kufema kwemasero. Chirevo chacho hachisi chenhema, asi zvachinoreva zvine muganhu kupfuura zvachinoratidzika kureva.

Zvinonyatsoyerwa nenhamba iri mutafura

Chaipimwa pakupedzisira kwaiva kushandiswa kweokisijeni, kuchiongororwa kuburikidza nekuongorora magasi. Harrop naBarron vakasefa ropa rakanga richangotorwa vachirishandisa donje pa10 °C kuti vabvise masero machena, vakazadza sampuli neokisijeni, vakaipatsanura, vakaisa dhayi mune chimwe chikamu, ndokuzoongorora zvikamu zvose zviri zviviri mushure mekuzvichengeta pakudziya kwakatarwa. Huwandu hwemasero machena hwaisawanzopfuura churu chimwe pa cubic millimetre. Kushandiswa kweokisijeni kwakataurwa sechikamu cheokisijeni yaiva musampuli pakutanga chakanga chanyangarika.

Vakayerawo carbon dioxide, asi ipapo nzira yekuyera yakavapa dambudziko. Ropa rakabviswa fibrin raiburitsa carbon dioxide inosvika mamirimole maviri pa100 mL muawa yekutanga yekuchengetwa pakudziya kwakatarwa, nezvikonzero zvakanga zvisinei nekuita kwemakemikari kwavaiongorora. Saka reshiyo dzekufema dzakabva muropa rose dzaisashandisika. Masero akagezwa chete, ari mumusanganiswa waLocke, ndiwo akapa nhamba dzaigona kududzirwa, uye idzodzo dzaiva pakati pe0.73 ne0.80, paine dhayi kana pasina.

Chinhu ichi hachisi chekungofuratirwa. Mumapepa aya, “kufema” kunoreva kuchinjana kwemagasi mukati memudziyo wakavharwa. Musiyano uripo pakati pekushandiswa kweokisijeni nekuburitswa kwesimba rinobatsira unoongororwa zvakasiyana muchii chinotaurwa nekushandiswa kweokisijeni pamusoro peATP uye chisingataurwi nazvo.

Dhayi yakawedzera zvakanyanya apo masero aifema zvishoma

Masero matsvuku eropa akakura emhuka dzinoyamwisa anenge asingashandisi okisijeni nenzira inogona kuyerwa. Masero matsvuku eropa eshiri ane nyukiriyasi anoishandisa. Harrop naBarron vakaedza marudzi ose ari maviri, uye musiyano wacho ndiwo musimboti wetsananguro yavo.

Musampuli dzemhuka dzinoyamwisa, dhayi yakaita kuti kushandiswa kweokisijeni kuwedzere kakapetwa makumi maviri kusvika makumi mashanu. Muropa rehanzi, rine masero akanga achitofema, haina kumbopfuura kuwedzera mwero wacho zvakapetwa kaviri. Kana kufema kwekutanga kwaiva kwakanyanya, dhayi yaigona kuwedzera zvishoma.

Musiyano iwoyo unoonekwawo mukati merudzi rumwe chete. Sampuli yemunhu aiva nepolycythemia vera, yaiva nemasero asati akura, yakashandisa 11.3 kubva muzana yeokisijeni yayo pasina dhayi, zvichienzaniswa ne2.8 kubva muzana pamasero akagezwa akatorwa kumupi weropa ane utano. Dhayi yakangoikwidza kusvika pa63.4 kubva muzana, kuwedzera kwakapetwa katanhatu apo masero akajairika akawedzera kanoda kusvika makumi matatu. Dhayi yakaita kuti masero akakura aite zvakafanana neasati akura, sezvakataurwa nevanyori kuti ndiyo nzira yayaishanda nayo.

Cyanide yaifanira kunge yazvimisa

Cyanide yaizivikanwa kuti inomisa oxidation mumasero. Kunyange zvakadaro, Harrop naBarron vakaiwedzera pamwe chete nedhayi.

Zvakawanikwa muropa remombe zvinomiririra zvimwe zvakawanikwa. Huwandu hweokisijeni hwakaderera kubva pa22.61 kusvika pa22.03 volumes per cent mumaawa matatu pasina chakawedzerwa. Pakaiswa methylene blue, hwakaderera kusvika pa12.84. Pakaiswa methylene blue nepotassium cyanide ine concentration yechikamu chimwe pazviuru zvishanu zvemolar, hwakaderera kusvika pa12.74. Kushanda kwedhayi kwakanga kusina kunyanya kukanganiswa.

Mufananidzo: chinyorwa chinotevedza kufema kudiki kwemasero matsvuku eropa remunhu nguva yose. Museve unoratidza pakawedzerwa methylene blue. Izvozvo zvisati zvaitika, mutsetse unenge wakati sandara; pashure pazvo unokwira zvishoma nezvishoma. Wakatorwa kubva kuna Harrop naBarron, Journal of Experimental Medicine 48:207–223 (1928), PMID 19869478.

Vakagumisa kuti dhayi yaishanda semutakuri weokisijeni, “ichipa chinhu chakanyangarika mumasero matsvuku eropa asina nyukiriyasi emhuka dzinoyamwisa dzakura.” Makore maviri gare gare, Michaelis naSalomon paRockefeller Institute vakatsanangura kusawirirana kwacho nenzira yakaramba iripo. Zvimwe kushanda kwedhayi kwaiva “chinhu chainoitwa nedhayi netsaona,” kana kuti dhayi yaiva “muenzaniso wechimwe chinhu chisingazivikanwi chiri muzvipenyu.” Sarudzo pakati petsananguro idzodzo haisati yagadziriswa. Chachinja ndechekuti masero acho ava kukwanisa kutsanangurwa zvakanyatsorurama.

Zvakatorwa muchiropa zvakaita zvimwe chetezvo

Michaelis naSalomon vakaratidza kuti zvinhu zvakatorwa munhengo dzemuviri pachishandiswa mvura inotonhora zvinokonzera kuwedzera kumwe cheteko. Zvakatorwa muchiropa chegonzo ndizvo zvakashanda zvakanyanya, zvichiteverwa neitsvo, spleen, nechende. Zvakatorwa muuropi, mutsandanyama, neserum yeropa zvakaita zvishoma kana kusaita chinhu.

Vakazoyedza zvakatorwa izvi vachishandisa carbon monoxide. Kufema kwakakonzerwa nezvakatorwa izvi kana nedhayi hakuna kuchinja mumusanganiswa une carbon monoxide inosvika zvikamu 98 kubva muzana uye okisijeni inosvika zvikamu 2 kubva muzana. Musanganiswa mumwe chete wemagasi wakadzivisa kufema kwembiriso nekufema kwemasero matsvuku eropa ehuku ane nyukiriyasi nezvikamu zvinenge 60 kusvika 80 kubva muzana, zvinoenderana netsananguro yaWarburg yechinhu chinokonzera kufema chinokanganiswa necarbon monoxide. Saka chinhu chemumatishu chisina kuzivikanwa ichocho chakanga chisiri enzyme yaWarburg.

Zvinhu zvakati wandei zvaifungidzirwa kuti ndizvo zvacho zvakazorambwa. Glutathione yaiva muchimiro chakareduced uye cysteine zvakangokonzera kushandiswa kweokisijeni yaidiwa neoxidation yazvo pachazvo. Coenzyme yaHarden naYoung yakarambwa nokuti tsandanyama ndiyo yaiva nayo yakawanda asi ndiyo yakaratidza kushanda kwaiva pakati pezvidiki zvikuru. Cytochrome yaKeilin, yakanga yapiwa zita makore mashoma akanga apfuura, yakasanganawo nekupokana kumwe cheteko. Pfungwa yavo pachavo, yekuti chinhu chinoshanda chaibva munyukiriyasi yesero, vakaiti yaingova pfungwa chete.

Kushanda kwacho kwaida mamwe mamiriro

Masero aifanira kunge akakwana, asina kuputika. Kuputswa kwemasero matsvuku eropa nemvura yakadistillwa kana nechando chaidzokororwa kwakamisa kupindura kwawo kudhayi nezvakatorwa munhengo dzemuviri. Kana mamwe chete emasero akaputswa, kupindura kwacho kwaiderera zvichienderana nechikamu chakaputswa.

Shuga yaidiwawo. Glucose, fructose, nesucrose zvakatsigira kufema kwakakonzerwa nedhayi; mannitol haina. Pasina shuga pakanga pasina kufema zvachose, uye dhayi yairatidzika kukuvadza masero panguva iyoyo, saka glucose yakawedzerwa awa imwe kana maviri gare gare haina kudzoreredza kufema. Vanyori vakafungidzira kuti dhayi yaikonzera oxidation yechimwe chikamu chechimiro chesero chaidiwa nesero.

Chiedza chakawedzera kushanda kwedhayi zvakanyanya, kunyange zvazvo kushanda kwayo kwaionekwa zvakajeka murima. Mamwe madhayi aigona kuitwa reduction akaita zvakasiyana: toluylene blue yakashanda zvakaenzana kana kupfuura methylene blue, asi indigo disulfonate yakaita zvishoma zvikuru. Hapana hukama hwakajeka hwakaonekwa pakati pesimba reoxidation-reduction redhayi nekukura kwekushanda kwayo. Vanyori vakati izvi zvaikonzerwa nekukwanisa kwayo kupinda nemumembrane uye nehuturu hwayo.

Kushanda kwacho kwaitevedza kuvirisa, kwete kufema

Ongororo yaBarron ya1930 yakabva pakushandisa masero ari mumvura ichienda pakushandisa zvidimbu zvematishu enhengo dzakajairika dzemakonzo nemamota akanga aisirwa mune mumwe muviri.

Mumatishu akanga asina aerobic glycolysis, methylene blue haina kuita chinhu kana kuti yakaderedza kushandiswa kweokisijeni zvishoma: muitsvo nezvikamu zvinenge 14 kubva muzana, murwatata nezvikamu zvinenge 14 kubva muzana, muchiropa nezvikamu zvinenge 5 kubva muzana. Mumatishu anovirisa paine okisijeni, yakawedzera kushandiswa kwayo: muspleen nezvikamu zvinenge 8 kubva muzana, muuropi nezvikamu zvinenge 13 kubva muzana. Kuwedzera mumamota kwaibva pazvikamu zvinenge 27 kubva muzana mucarcinoma yezamu remunhu kusvika pazvinopfuura zvikamu 80 kubva muzana muRous sarcoma yehuku, zvichida kuenzana neaerobic glycolysis yetishu rimwe nerimwe.

Mufananidzo: poinzi imwe neimwe inomiririra rudzi rumwe rwetishu. Akisi yakatambarara inoratidza aerobic glycolysis; akisi yakamira inoratidza shanduko yekushandiswa kweokisijeni mushure mekuiswa methylene blue. Data rinobva kuna Barron, Journal of Experimental Medicine 52:447–456 (1930), Tafura 6, PMID 19869777.

Kuyedza necyanide kwakazoburitsa zvakanga zvakapesana nezvakanga zvamboonekwa. Apo kufema mumatishu aya akanga asingapinduri kwakavharwa necyanide uye kuvirisa kwakatanga kuoneka, dhayi yakawedzera zvakare kushandiswa kweokisijeni: muitsvo nezvikamu zvinenge 59 kubva muzana, muchiropa nezvikamu zvinenge 19 kubva muzana, muspleen nezvikamu zvinenge 286 kubva muzana, muchende nezvikamu zvinenge 200 kubva muzana.

Barron akapa pfungwa yekuti dhayi yaigona kushandiswa kuyedza simba rekuvirisa resero kana retishu, ndokupedzisa nefungidziro: kukurisa mamota kwezvizvarwa zvakawanda mumusanganiswa une dhayi kungazopedzisira kwaita kuti metabolism yebundu igare yakafanana neyematishu akajairika. Uyu waiva mubvunzo wekutsvakurudza wakanyorwa muna 1930. Hachisi chirevo chekuti inzira yekurapa.

Zvinhu izvo kuyedza uku kwakanga kusingagoni kunge kuri kuyera

Muna 1928 na1930, “kufema” kwaireva kushandiswa kweokisijeni. Adenosine triphosphate yakatanga kutaurwa muna 1929, uye kuratidzwa kwekuti mitochondria dzakaparadzaniswa dzinobatanidza phosphate dzichitakura maelectrons kwakaitika muna 1949. Hapana rimwe remapepa ekutanga emethylene blue raigona kunge rakayera kugadzirwa kwesimba, uye hapana rakati rakanga razviita.

Masero ndiwo dambudziko rechipiri. Masero matsvuku eropa akakura emhuka dzinoyamwisa haana mitochondria. Panguva yaanenge ari mareticulocyte, anobvisa mitochondria nemaribosome kuburikidza neautophagy inosarudza zvinobviswa. Ndokusaka sero dzvuku reropa rakakura richishandisa glycolysis uye richivimba zvakanyanya nepentose phosphate pathway yaro kuti riwane NADPH, kwete ATP. Mukuyedza kwa1928, makanga musina cheni yekutakura maelectrons yaigona kugadziriswa. Chero chinhu chaishandisa okisijeni chakanga chichiita chimwe chinhu.

Chimwe chinhu ichocho chava kutsanangurwa zvakati tsvikiti. Methylene blue inogamuchira maelectrons kubva kuzvinhu zvesero zvinopa simba rereduction, yoita leucomethylene blue, iyo inozoitwa oxidation zvakare neokisijeni yemamorekuru, ichipedzisa kutenderera kwacho. Mumasero matsvuku eropa remunhu, flavin reductase inotsamira paNADPH nemadisulfide reductase ane hukama nayo ndiwo anoita danho rereduction. Ongororo ya1976 yakashandisa glucose yakaiswa chiratidzo yakawana kuti dhayi inomutsa hexose monophosphate shunt, kuti kushanda kwayo paconcentration yakaderera kwaitsamira paoxyhemoglobin, uye kuti concentration yakakwirira, 0.1 mM, yaigadzira hydrogen peroxide. Dhayi yaitwa reduction painosangana neokisijeni inogadzira peroxide, kwete zvisungo zvephosphate.

Mumitochondria, tsananguro yacho ine kupokana kwakawanda kupfuura zvinobvumwa netsananguro dzinonyanya kupararira. Rimwe boka retsvakurudzo rinoti dhayi inotora maelectrons munharaunda yecomplex I yoaendesa kucytochrome c. Rimwe rakawana kuti mumitochondria yeuropi hwemakonzo madiki haigoni kudzoreredza kufema mushure mekuvharwa neantimycin, rikaisa chinogamuchira maelectrons panzvimbo inosungirirwa ubiquinol pacomplex III. Rechitatu rakawana kuti concentration yakaderera yakawedzera kushandiswa kweokisijeni pakuzorora asi ichisiya kufema kunomutswa neADP kusina kuchinja. Chirevo chinonyatsobata pfungwa yacho ndechekuti kushandiswa kweokisijeni kunokonzerwa nemethylene blue hakutombotsamiri paoxidative phosphorylation: kwakaramba kuchiitika apo cheni yekufema yakadziviswa, uyewo mumasampuli akanga asina cheni yacho.

Kuyera kwakananga kwemusiyano pakati pezviyero zviviri izvi kuripowo. Mumitochondria dzakaparadzaniswa dzechiropa chegonzo, kuwedzera concentration yedhayi kwakaderedza respiratory control ratios ndokukonzera kuzvimba, izvo vanyori vakatsanangura sekupatsanurwa kwemaitiro akabatana muoxidative phosphorylation. Muchiropa chegonzo chaipfuudzwa mvura nemumidziyo yeropa, methylene blue yakawedzera kushandiswa kweokisijeni asi reshiyo yeATP kuADP ichiderera.

Zvinyorwa zvekurapa zvinopa pfungwa imwe cheteyo. Methemoglobinemia inorapwa nemethylene blue inokwana mirigiramu imwe kusvika mbiri pakirogiramu, ichiiswa mutsinga, kuburikidza nenzira yereduction inotsamira paNADPH, uye huwandu hwakakura hunokonzera chirwere chacho chiri kurapwa. Ongororo imwe cheteyo inocherechedza kuti kushanda kwayo semushonga unorwisa chepfu kwakanyanya mumasero matsvuku eropa asina kuputika, uye kunoderera kana kuputswa kwemasero kuripo. Izvi ndizvo zvakawanikwa pamusoro pekuputswa kwemasero muna 1930 zvichionekazve makore makumi manomwe nematatu gare gare muchipatara. Chinhu chine kushanda kunobatsira nekushanda kunokuvadza kumativi akasiyana emuganhu wehuwandu hachisi chinhu chine huwandu hunofanira kufungidzirwa kubva pakuyera kushandiswa kweokisijeni.

Nei batch ichifanira kubatanidzwa muhurukuro

Harrop naBarron vakacherechedza chimwe chinhu chinokodzera kuverengwa zvakare. Methylene blue pachayo inogadzira bhandi muspectroscope pakati pemabhendi maviri anozivikanwa nemethemoglobin, “izvo zvinogona nyore kukonzera kuvhiringidzika pakungoongorora usina kunyatsotarisa.” Dhayi inogona kutevedzera chinhu chiri kutsvakwa.

Dambudziko rimwe chete iri rine chimiro chemakemikari. Phenothiazine ine hukama nayo inowanikwa sechikamu chidiki ine maitiro ayo eredox, kukwanisa kwayo kupinda nemumembrane, uye huturu hwayo. Mumwe muenzaniso wakanyorwa unoratidza zvakajeka kukosha kwazvo: muongororo yekuisa iodine mumethylene blue, sampuli dzaiva nezvikamu zvinenge 7 kusvika 8 kubva muzana zveAzure B dzakagadzira zvikuru Azure B yakaiswa iodine. Saka tsvina yacho, kwete methylene blue, ndiyo yakatonga kuita kwemakemikari.

Chero munhu anoshandisa methylene blue sereagent, murabhoritari, pabhenji rekudhaya masampuli, kana mutangi rehove, ari kushanda nechinhu chisina kutsanangurwa kunze kwekunge batch yacho yaongororwa kuti zvairi zvizivikanwe. Zvinhu zvine hukama nayo, kuzivikanwa kwechinhu, tsvina yezvinhu zvemakemikari, zvisaririra zvezvinonyungudutsa, uye zvisaririra mushure mekupiswa chikamu chekuyedza pachako, kwete kungori mapepa anoperekedza kuyedza. Blupreme inowana methylene blue yayo kubva kumugadziri wemishonga uye inoburitsa chitupa chakazara chekuongorora, ichiyedza zvose zvinodiwa neUSP pane kungoyedza simbi dzinorema chete; gwaro rekuverenga chitupa chekuongorora uye zvinosimbiswa nekudanwa kuti USP-grade nezvisingasimbiswi nazvo zvinotsanangura maitiro ekuenzanisa gwaro nechirevo chiri kuitwa.

Nhamba yacho yaiva yakarurama

Kuyedza kwa1928 na1930 kwakaitwa nokungwarira, uye zvavakanyanya kuona zvakaramba zvichitsigirwa: dhayi inogona kuitwa reduction inogona kuita kuti sero risinganyanyi kufema rishandise okisijeni pamwero wakapetwa kakawanda pane waro wekuzorora, risingapfuuri nedanho rinovharwa necyanide.

Kukanganisa kunozooneka gare gare, apo “kufema” kunodudzirwa nezvakunoreva mazuva ano. Kuyera kuchinjana kwemagasi kunokuudza kuti maelectrons akasvika kuokisijeni. Hakukuudzi kuti mamwe acho akashandiswa kugadzira ATP, uye musero risina mitochondria musiyano pakati pezvirevo zviviri izvi wakakura zvikuru. Mufananidzo wakafara wezvinoitwa nemethylene blue mumasero unoisa musiyano mumwe chete uyu mune mamiriro akafara, uye nhoroondo yechinhu ichi kubva pakuva dhayi kusvika pakuva mushonga inotsanangura kuti chinhu chekupa micheka ruvara chakasvika sei pakushandiswa mukuyedza uku pakutanga.