Nkhani
Methylene blue mu mayeso a G6PD a labotale: zimene utotowu ukuchita kwenikweni
Wophunzira akuyang'ana machubu awiri atawasunga pa kutentha koyenera kwa nthawi inayake. Limodzi lasintha n'kukhala ndi mtundu woyembekezeredwa. Lina silinasinthe. Wophunzirayo akuganiza kuti wodwala wachiwiriyo ali ndi kuchepa kwa glucose-6-phosphate dehydrogenase. Maganizo amenewo angakhale olondola, koma njira yake yoganizira ikudumpha mfundo imene imapangitsa mayesowa kugwira ntchito. Palibe chimene chili m'chubumo chimayeza enzayimuyo mwachindunji. Chubucho chimayeza zimene enzayimuyo imatulutsa, pogwiritsa ntchito methylene blue ngati mkhalapakati.
Njira imeneyi yoyezera mosalunjika ikamveka bwino, ndondomeko, zoyerekezerera, ndi mndandanda wautali wa zotsatira zolakwika sizionekanso ngati zinthu zopanda chifukwa.
Yankho lachidule
Methylene blue pa mayeso a G6PD ndi chizindikiro cha redox. Enzayimu ya G6PD imapanga NADPH, ndipo machitidwe amene amadalira NADPH amachepetsa methylene blue ya buluu mwa kuipatsa maelekitironi kuti isanduke mawonekedwe ake a leuco opanda mtundu (kapena amaigwiritsa ntchito kunyamula maelekitironi). Ntchito yabwinobwino ya enzayimu imapangitsa kuchepetsedwa kumeneku kuchitika bwino. Ntchito yochepa imapangitsa kuchepetsedwako kukhala kochedwa kapena kosakwanira. Chimenecho ndicho chizindikiro chonse cha mayesowo.
Njira ziwiri zakale zimagwiritsa ntchito utoto womwewo koma zimasiyana pa chimene chimayang'aniridwa pamapeto. Kuzisiyanitsa kumachotsa chisokonezo chochuluka.
Mayeso awiri, utoto umodzi, zoyang'aniridwa zosiyana pamapeto
Yoyamba ndi mayeso ochepetsa methylene blue mwachindunji, amene anafotokozedwa ndi Sass ndi anzake ndipo kenako anawunikidwa ndi Gibbs. Apa chimene chimayang'aniridwa pamapeto ndi utoto womwewo: methylene blue ya buluu yomwe ili mu mawonekedwe a oxidation imachepetsedwa n'kuyandikira leucomethylene blue yopanda mtundu, ndipo liwiro la kusinthako limasonyeza mphamvu yopanga NADPH. Pepala loyambirira lofotokoza kuunikaku lalembedwa pa PubMed pansi pa Sass ndi anzake (1966).
Yachiwiri ndi mayeso a Brewer ochepetsa methemoglobin. Poyamba sodium nitrite imapangitsa hemoglobin kukhala methemoglobin yabulauni kudzera mu oxidation. Kenako methylene blue imagwira ntchito ngati mkhalapakati posamutsa maelekitironi: methylene blue yochepetsedwa imapereka mphamvu yochepetsa ku methemoglobin, n'kuisandutsa hemoglobin yofiira. Choncho chimene chimaoneka pamapeto ndi bulauni poyerekeza ndi wofiira, osati buluu poyerekeza ndi kusowa kwa mtundu. Njira yoyambirira inasindikizidwa mu Bulletin ya Bungwe la Zaumoyo Padziko Lonse (Brewer ndi anzake, 1960).
Kunena kuti mayeso a Brewer ndi mayeso ochotsa mtundu wa utoto si kulondola kwenikweni pa nkhani ya kemistri. Utoto umayenda mobwerezabwereza pakati pa mawonekedwe ake; hemoglobin ndiyo imasintha mtundu. Chithunzi chili m'munsimu chikutsatira njira zonse ziwiri.
Kusintha kwa mankhwala kumene kuli maziko ake
G6PD imafulumizitsa sitepe yoyamba ya oxidation mu njira ya pentose phosphate:
Glucose-6-phosphate + NADP+ becomes 6-phosphogluconolactone + NADPH + H+
Maselo ofiira okhwima amadalira kwambiri njirayi kuti apeze NADPH, monga tafotokozera mu Blood ndi Luzzatto ndi anzake (2020). NADPH imasunga glutathione mu mawonekedwe ochepetsedwa ndipo imateteza selo ku kuwonongeka kochitika chifukwa cha oxidation. Methylene blue imalowa m'dongosolo limeneli ngati cholandira maelekitironi. G6PD yokwanira imapereka NADPH yokwanira, yomwe imapangitsa kuchepetsedwa kuchitika bwino. G6PD yochepa imadula unyolowu pa cholumikizira chake choyamba.
Ganizirani utotowu ngati chipangizo chosonyeza kuchuluka kwa mafuta, osati injini. Umanena za kupezeka kwa NADPH. Chilichonse china chimene chimasintha kupezeka kwake, kapena mmene zotsatira zimawerengedwera, chingasinthe chizindikiro popanda kusintha kulikonse kwa jini.
Mmene ndondomeko ya mtundu wa Brewer imayendera
Njira zosakanizira zinthu za mayeso zakhala zikusiyana kwa zaka zambiri, choncho kufotokoza kophunzitsira sikuyenera kuganiziridwa ngati ndondomeko yoyezera odwala. Labotale iliyonse imatsatira ndondomeko yake yokhazikika yogwirira ntchito yomwe yatsimikiziridwa. Poganizira chenjezo limeneli, njira yakale yogwiritsa ntchito machubu imagwira ntchito motere, monga zalembedwera pogwiritsa ntchito machubu atatu ndi kuwasunga pa kutentha koyenera kwa maola 3 mu njira iyi yosindikizidwa:
- Chubu cha wodwala: magazi athunthu pamodzi ndi glucose, sodium nitrite, ndi methylene blue. Nitrite imapanga methemoglobin, glucose imathandiza kagayidwe ka zinthu m'maselo ofiira, ndipo utoto umapereka njira yonyamulira maelekitironi yodalira NADPH.
- Choyerekezerera cha oxidation: magazi pamodzi ndi nitrite ndi glucose koma opanda methylene blue. Chimasonyeza mtundu wabulauni wa methemoglobin pamapeto, womwe ndi chizindikiro cha kuchepa.
- Choyerekezerera cha hemoglobin: magazi pamodzi ndi madzi. Chimasonyeza mtundu wofiira wa hemoglobin pamapeto, womwe ndi chizindikiro cha mkhalidwe wabwinobwino.
- Sungani pa kutentha koyenera machubu onse pafupi ndi 37 C, nthawi zambiri kwa maola 3, mukumasakaniza pang'onopang'ono.
- Chepetsani kuchuluka kwake poika gawo laling'ono lochokera m'chubu chilichonse m'madzi oyeretsedwa ndi distillation kuti maselo aphwanyike komanso kuyerekeza mitundu kukhale kosavuta.
- Yerekezerani chubu cha wodwala ndi zoyerekezerera ziwirizo. Kubwerera ku mtundu wofiira kumasonyeza mphamvu yaikulu yochepetsa. Kupitiriza kukhala pafupi ndi bulauni kumasonyeza kuti pangakhale kuchepa kwakukulu.
Kusakaniza kumodzi kwakale kunkagwiritsa ntchito magazi pafupifupi 1 mL ndi zinthu za mayeso zokhala pafupifupi 180 mM sodium nitrite, 280 mM dextrose, ndi 0.4 mM methylene blue. Onani manambala amenewo ngati chitsanzo chochokera m'mabuku a sayansi, osati muyezo wogwiritsidwa ntchito kulikonse.
Kuwerenga zotsatira
| Mkhalidwe | Maonekedwe pa mayeso a Brewer | Tanthauzo |
|---|---|---|
| Wabwinobwino | Chubu cha wodwala chimayandikira mtundu wofiira wa choyerekezerera cha hemoglobin | Mphamvu yochepetsa ili pamwamba pa malire a kuunika |
| Wochepa | Chubu cha wodwala chimakhalabe chabulauni, pafupi ndi choyerekezerera cha methemoglobin | Kuunika kumasonyeza kuti pangakhale kuchepa kwakukulu kwa G6PD |
| Wapakatikati | Mtundu uli pakati pa mitundu ya zoyerekezerera | Musatchule mkhalidwe wa phenotype; yezani kuchuluka kwa ntchito ya enzayimu |
Kuunika kosapereka kuchuluka kumasiyanitsa bwino kwambiri pafupi ndi 30 mpaka 40 peresenti ya ntchito yabwinobwino, monga tafotokozera mu ndemanga iyi ya Malaria Journal yokhudza mipata pozindikira G6PD. Choncho zotsatira zabwinobwino za kuunika zimatanthauza kuti mayesowo sanazindikire kuchepa kwakukulu. Sizitsimikizira kuti ntchito ya enzayimu ndi yabwinobwino kwathunthu.
Chifukwa chake kuunika kungasocheretse
Kuchuluka kwa ma reticulocyte pambuyo pa hemolysis ndi chifukwa chodziwika bwino cha zotsatira zosonyeza zabwinobwino molakwika. Maselo ofiira atsopano ndi ma reticulocyte amakhala ndi G6PD yochuluka kuposa maselo akale. Pa nthawi ya hemolysis yadzidzidzi, maselo akale kwambiri komanso amene ali ndi kuchepa kwakukulu angakhale atawonongeka kale, n'kusiya maselo atsopano amene ntchito yawo imayesedwa n'kuoneka yabwinobwino. Zimenezi zimasocheretsanso mayeso oyeza kuchuluka, osati kuunika pogwiritsa ntchito utoto kokha.
Kuikidwa magazi posachedwapa kumabisa kuchepa chifukwa maselo ofiira a munthu wopereka magazi amawonjezera enzayimu yawoyawo ku chitsanzocho. Mayo Clinic Laboratories imatchula kuikidwa magazi pakati pa zifukwa za zotsatira za enzayimu zosadalirika, pamodzi ndi kuchuluka kwa ma reticulocyte ndi kusokoneza kochokera ku maselo oyera a magazi.
Akazi okhala ndi ma aleli osiyana a jini ya G6PD ndiwo ovuta kwambiri kuwunika. Kuzimitsidwa kwa kromozomu ya X mwachisawawa kumapanga kusakanikirana kwa maselo ofiira abwinobwino ndi amene ali ndi kuchepa, ndipo ntchito yonse ya enzayimu ingakhale paliponse kuyambira yochepa kwambiri mpaka yooneka yabwinobwino. Kuunika kosapereka kuchuluka nthawi zambiri sikuzindikira mikhalidwe ya phenotype yapakatikati. Zolemba za WHO zokhudza mayeso a G6PD zimafotokoza mwatsatanetsatane cholepheretsa chimenechi.
Nthawi imene chitsanzo chakhala chilipo, kutentha posunga, kusiyana kwa mmene anthu amawerengera mtundu womaliza ndi maso, hematocrit, ndi ubwino wa zinthu za mayeso zimawonjezera kusokonezeka kwa kusanthula pa zotsatira za zinthu zachilengedwe zimenezi.
Kuunika si kutsimikizira mkhalidwe
Zotsatira za kuunika zosonyeza kuchepa, mkhalidwe wapakatikati, kapena zosagwirizana ndi zimene zikuoneka kwa wodwala zimafuna mayeso oyeza kuchuluka kwa ntchito ya enzayimu ya G6PD. WHO imaona kuyeza ndi spectrophotometry pa kutentha kolamulidwa ngati njira yoyerekezerera. Pa mayesowo, chitsanzo cha maselo ofiira ophwanyidwa chimapatsidwa glucose-6-phosphate ndi NADP+, ndipo NADPH yomwe imapangidwa imatsatiridwa ndi kuwonjezeka kwa kuyamwa kwa kuwala pa 340 nm. Liwiro la kusintha limayenderana ndi ntchito ya enzayimu, ndipo nthawi zambiri limawerengedwa mogwirizana ndi hemoglobin ngati mayunitsi pa gramu. Ntchito zamakono za labotale yoyezera odwala zafotokozedwa mwachidule mu zolemba za m'ndandanda wa mayeso a Mayo zokhudza ntchito ya G6PD.
Kutanthauzira kumagwiritsa ntchito malire oyerekezerera otsimikiziridwa ndi labotale, amene nthawi zambiri amaikidwa mogwirizana ndi ntchito yabwinobwino ya enzayimu m'gulu la anthu, ndipo kuchepa nthawi zambiri kumafotokozedwa pafupi ndi malire a 30 peresenti, pomwe ntchito yapakatikati imakhala pamwamba pake. Zofunikira za WHO pa zinthu zopangira mayeso a G6PD (2022) zimafotokoza dongosolo limenelo lotsimikizira mkhalidwe. Ngati hemolysis kapena kuikidwa magazi posachedwapa kumapangitsa kudziwa phenotype kukhala kosadalirika, mayeso abwerezedwe maselo ofiira akabwereranso pa mkhalidwe wolinganizika, ndipo mayeso a mamolekyulu angawonjezere mfundo popanda kulowa m'malo mwa kuyeza enzayimu.
Kusiyanitsa koyenera kusunga
Methylene blue pa mayesowa ndi chinthu cha mayeso a labotale chogwiritsidwa ntchito kunja kwa thupi. Mfundo imeneyi ndi yosiyana ndi nkhani ya chitetezo popatsa wodwala methylene blue, yomwe yafotokozedwa mu malangizo okhudza nthawi zimene methylene blue siyenera kugwiritsidwa ntchito. Kemistri ya m'thupi imalumikiza nkhani ziwirizi, chifukwa mankhwalawo amadaliranso kuchepetsedwa kodalira NADPH, koma utoto wa m'chubu cha mayeso si mlingo wa mankhwala m'magazi. Kuti mumvetse mmene utotowu umagwirira ntchito popaka maselo mtundu pa ntchito ina, onani mfundo zopakira mtundu ndi methylene blue, komanso za gulu la mankhwala limene uli mmenemo, utoto wa redox mu labotale.
Kuunika kumafunsa ngati mphamvu yochepetsa ikuoneka yosawonongeka. Kutsimikizira kumafunsa kuchuluka kwa enzayimu yomwe ilipo kwenikweni. Utotowu umayankha bwino funso loyamba, bola ngati palibe amene akuufuna kuti uyankhe lachiwiri.