Te hanga ānau whakatikatika mō te kikorangi metilīni: ngā tauira pātea, te awhe, me te rārangitanga

Ka whakamahi tahi ngā kaitātari tokorua i te pounamu kotahi o te wairewa pūtake kikorangi metilīni, ā, ka hanga ia tangata i tāna ake ānau whakatikatika. Ka whiwhi te tuatahi i tētahi rārangi tōtika mā te pūtake kore, ā, ka ine māia i ngā tauira kāore anō kia mōhiotia te kukū. Ka whiwhi te tuarua i tētahi rārangi e kōpiko ana i tōna pito runga, tētahi haukoti kāore e tata ki te kore, me ngā hua tauira e neke ana i te waimehatanga. He ōrite te waikano, te taputapu, me te roangaru. Ehara te pūkenga i te take o te rerekētanga. Ko te hoahoa kē: he aha ngā matū i roto i te tauira pātea, ko ēhea kukū i kōwhiria, ā, i noho rānei te awhe i tirohia ki roto i te wāhanga e tika tonu ai tētahi tauira pāngarau māmā.

He kirimana te ānau whakatikatika i waenga i te urupare a te taputapu me te kukū, ā, ka whai mana anake i raro i ngā āhuatanga i hangaia ai. Mō te kikorangi metilīni, he wāhi ngoikore kua mōhiotia tō taua kirimana. Ka hono te waikano ki a ia anō i roto i te wai, nō reira ka rerekē te momo matū ngongo i te rerekētanga o te kukū. E kore tētahi ānau i hangaia puta noa i taua rerekētanga e tōtika, ā, ahakoa pēhea te nui o te tātari tauanga, e kore e whakatikaia. E whakaatu ana tēnei tuhinga me pēhea te hoahoa i te ānau kia whai tikanga ai te rārangi: kōwhiria te tauira pātea tika, puritia te awhe ki roto i te rohe monomere, mahia ngā tāruarua pono, ā, whakaarohia te kōpikopiko hei mōhiohio, kaua hei haruru.

Tīmata ki te tauira pātea, kaua ki ngā tauira paerewa

Mā te tauira pātea e tautuhi te kore, ā, mō te kikorangi metilīni me whai i ngā mea katoa hāunga te waikano. Kia ōrite te kōeke wai, te pūreirei me te pH, ngā tote, te hautanga whakarewa āpiti, me te pāmahana. Whakaritea ngā tauira whakatikatika i roto i taua taiao matū ōrite. He take matū tēnei, ehara i te take whakahaere noa. Ka neke te tote me te whakarewa i te taurite i waenga i te monomere, te dimere, me ngā huinga nui ake, ā, ka huri te tūāwhiorangi i te roangaru ine. Kāore e taea e tētahi tauira pātea wai-anake te whakatika i te rerekētanga o ngā momo matū nā te taiao matū i hua ai. I roto i te tātari whānui o ngā tūāwhiorangi kikorangi metilīni i te wai, e wātea ana mā tōna pūkete uru tuwhera, i whakarerekētia te konutai pūhaumāota mai i te 0 ki te 0.15 M nā te mea ka neke te kaha katote i te taurite, ā, ka piki te whakahuihui i te tāpiritanga tote.

Kotahi te putanga ā-mahi: kaua rawa e whakatikatika ki te wai kua tangohia ngā katote, kātahi ka ine i ngā tauira kei roto i te pūreirei pākawa tūtae, i te wai para kua tātarihia rānei. Ki te kore he taiao matū e wātea ana kāore he matū tātari i roto, he pai ake te tāpiri paerewa ki te tauira tonu i tētahi ānau ā-waho kāore e ōrite ana te taiao matū. Kei te kōrero mō te taiao matū me ngā whakararuraru ētahi atu aratohu hei pupuri i te pono o ngā whakamātautau wai, ā, kei te tirohanga whānui mō te hoahoa whakamātautau ngā mātāpono whānui o ngā whakamātautau ine-rahinga waikano.

Kōwhiria he awhe e noho matua ai tētahi momo matū kotahi

Ko te ine-rahinga i te pae roangaru roa e tata ana ki te 664 nm e whakapae ana he monomere te nuinga o te waikano. I roto i te wai noa, ka tika noa taua whakapae i te wā e noho waimeha ana ngā wairewa, arā, kei te rohe micromolar iti. E whakaatu ana ngā rōnaki i whakaputaina mai i ngā inenga i te paenga: ko ngā whakatikatika i puritia ki raro iho i te āhua 7 micromolar i roto i ngā pūtau 1 cm ka whiwhi rōnaki e tata ana ki te 82,000 M-1 cm-1, engari ko tētahi whakatikatika i whakawhānuitia ki te āhua 37 micromolar ka heke ki te āhua 67,100, nā te mea kua nui kē te dimere i roto i ngā tauira paerewa o runga. Kua tapaina ngā āhuatanga ine o ia whakarea e whakahuatia ana i konei ki te tohutoro tūāwhiorangi mō te kikorangi metilīni, koia te wāhi tika hei whakataurite i ngā tau; e whakamahia ana aua tau i tēnei tuhinga hei tautuhi noa i te awhe.

Ka ahu tika mai ngā ture hoahoa:

1. Kia waimeha tonu te tauira paerewa teitei rawa, kia pūmau ai te hanga o te tūāwhiorangi. I te wā whakawhanake, matawaitia te awhe katoa o te rama kitea (tata ki te 550 ki te 750 nm) mō ia taumata, kaua e pānui i te roangaru kotahi anake. Ki te rerekē te hanga o ngā tūāwhiorangi kua whakaritea ki tētahi tauine kotahi i te rerekētanga o te kukū, kei te huri te matū, ā, he whānui rawa te awhe.
2. Aroturukitia te ōwehenga o te ngongotanga e tata ana ki te 610 nm ki te ngongotanga e tata ana ki te 665 nm puta noa i ngā taumata. Ki te piki taua ōwehenga i te pikinga o te kukū, e tipu ana te pakihiwi dimere, ā, kei te kore e tika te tauira pāngarau monomere. Ka whakamāramatia anō tēnei tohu i roto i te aratohu mō te whakahuihui me ngā pānga o te kukū.
3. Whakamahia he pūtau 1 cm mō ngā mahi micromolar iti. Ki te tata te rōnaki ki te 82,000, ka puta te ngongotanga tata ki te 0.41 i te 5 micromolar, ā, ka puta te āhua 0.82 i te 10 micromolar; e pai ana ēnei mō te nuinga o ngā taputapu.
4. Mō ngā wairewa pūtake kukū, whakapotoa te ara, kaua e whakaae ki te ngongotanga kei tawhiti ake i te 1. Engari kia mahara, mā te ara poto e whakatika te ine rama, kaua te matū. Ka whakawhirinaki te whakahuihui ki te kukū, kaua ki te roa ara, nō reira me whai tonu te wairewa pūtake kukū i tōna ake tukanga e whai whakaaro ana ki ngā momo matū, kaua i tētahi ānau monomere waimeha.

I roto i te whakahāngaitanga whānui e tautoko ana i te nuinga o tēnei whakamārama, i whakamahia ngā tūāwhiorangi 224 mai i te 1.1 micromolar ki te 3.4 millimolar, ā, i kitea ka taea kē te kite i te tohu tetramere i raro iho i te 34 micromolar. Ko te akoranga o taua kitenga: he whāiti ake te awhe rārangitanga i tā te whakaaro noa e tohu ai, nō reira manatokoa, kaua e whakapae noa.

Ngā taumata, ngā tāruarua, me te raupapa

Kāore tētahi ānau e toru noa ōna ira e whakamana i tētahi mea. E tono ana ngā aratohu whakamana o ICH Q2(R2) kia kaua e iti iho i te rima ngā kukū hei aromatawai i te rārangitanga, ā, mā te ono ki te waru ngā taumata ehara i te kore e pono ake ai te ānau mahi, otirā ki te pito runga e tīmata ai te whakahuihui. Whakaritea ngā taumata kia kapi ai te awhe tauira e hiahiatia ana, me te whakapiki i te maha o ngā taumata ki ngā wāhi e tino tūpono ai te kōpikopiko; kaua e whakamahi i tētahi raupapa āhuahanga i kōwhiria mō te ngāwari noa.

Inea ia taumata kia rua, kia toru rānei, mā ngā whakaritenga motuhake, kaua mā te pānui kia toru ngā wā i te ipu kotahi. Mā ngā tāruarua motuhake e whakaatu ngā hapa whakarite; mā ngā pānuitanga tāruarua o te ipu kotahi e whakaatu noa te haruru o te taputapu. I te rangahau whānui o 2020, i inea ia wairewa kia toru ngā wā, ā, he tikanga tīmatanga pai tonu tēnā. Me matapōkere te raupapa pānui, kia kore ai te paheke haere, te piri ki te mata karāhe, te panoni pāmahana rānei e āhua rite ki te kōpikopiko. I kitea e tētahi rangahau ipu ine kiripaka he tata ki te 3 ōrau o te waikano i piri ki te mata mai i tētahi wairewa 34 micromolar i roto i te 10 meneti, nō reira kia ōrite tonu te wā pā, te ipu ine, te horoi, me te raupapa puta noa i te raupapa ine.

Tauira tātai hei whakamārama (ehara i te raraunga ine)

He tau tito ngā tau kei raro nei hei whakaatu i te tātai. Ehara i te inenga taiwhanga, ā, kaua e whakahua hei raraunga kikorangi metilīni. Ko te mea nui ko te tauira o ngā arowhai e karapoti ana i aua tau.

Me whakaaro e ono ngā tauira paerewa kei roto i te pūreirei ōrite, he ara 1 cm, ā, ka pānuihia i te 665 nm mā te whakataurite ki te tauira pātea ōrite:

Mā te whakahāngai rārangi tōtika i te ngongotanga toharite ki te kukū ka whiwhi rōnaki e tata ana ki te 0.0805 waeine ngongotanga mō ia micromolar, me tētahi haukoti kei roto i ētahi waeine ngongotanga haumano ruarua o te kore. Mā te whakawehe i te rōnaki ki te roa ara ka huri hei āheinga ngongo ā-kitea: ko te 0.0805 mō ia micromolar e ōrite ana ki te āhua 80,500 M-1 cm-1, e hāngai ana ki tētahi awhe e noho matua ai te monomere i roto i te wai. Ehara ko te R pūrua te arowhai matua, ka tino pai hoki tōna āhua ahakoa pēhea. Ko ngā mea matua ko te haukoti tata-kore, te ōwehenga A610/A665 pūmau puta noa i S2 ki S5, me ngā toenga e marara ana kāore he ia. Ko te pikinga iti o te ōwehenga i S6 te tohu pono kia whakarerea taua taumata, kia tohua rānei te 7 micromolar hei paenga runga rangitahi mō tēnei huinga taiao matū me te taputapu.

Ka taea te huri tōtika i tētahi tauira kāore anō kia mōhiotia te kukū, i pānuihia hei 0.245 i raro i ngā āhuatanga ōrite: ko te 0.245 ka whakawehea ki te 0.0805 mō ia micromolar ka puta te āhua 3.0 micromolar, i mua i te whakamahi i tētahi tauwehe waimehatanga tauira. Pūrongoa te hua me ōna āhuatanga: te roangaru, te ara, te taiao matū o te tauira pātea, te awhe whakatikatika, me te rā.

Pepa mahi whakatikatika hei whakakī

Tāruatia tēnei tauira ki te pūkete taiwhanga mō ia taiao matū hou. Kotahi te taiao matū, kotahi te pepa, kotahi te ānau.

He take nui tō te rārangi mō te āhua o te tauhohe. Tērā pea he tote kore-wai te kikorangi metilīni hokohoko, he āhua whai-wai rānei pēnei i te āhua wai-toru, ā, mā te papatipu mōra hē e whakaharatau puku ngā kukū whakaingoa katoa. Kei te aratohu kounga me te kōeke ngā arowhai mō te parakore me ngā whakaritenga o te matū tīmatanga.

Manatoko mā tētahi tauira paerewa arowhai motuhake

Kaua rawa te ānau whakatikatika e māka i āna ake mahi. Whai muri i te whakahāngaitanga, whakaritea tētahi tauira paerewa arowhai mai i tētahi wairewa pūtake i paunatia motuhaketia, ki tētahi kukū e tata ana ki te pokapū o te awhe i whakaaetia, ā, pānuihia hei tauira kāore anō kia mōhiotia te kukū. Ki te ōrite i roto i ētahi ōrau ruarua, ka whakamanatia te raupapa whakarite; ki te kore e ōrite, ka tohu tēnā ki te wairewa pūtake, ki ngā waimehatanga rānei, kaua ki te taputapu. Whakahokia te tauira paerewa arowhai mō ia rōpū tauira. Ki te paheke tōna hua, engari kāore te ānau i rerekē, whakapaea ngā wairewa mahi, te ipu ine rānei, i mua i ngā tauira. Mā tēnei tikanga kotahi e kitea ai te nuinga o ngā hapa huna: te whakamahi i te papatipu mōra whai-wai mō tētahi pounamu kore-wai, tētahi pipeta moroiti kāore i te tika tōna whakatikatika, tētahi pūreirei i whakaritea ki te kaha katote hē. Tuhia te ōwehenga o te hua arowhai ki te uara e tūmanakohia ana ki te pepa mahi, ki te taha o te rōnaki, kia kite ai te kaipānui o muri mai i whakamātauria te rārangi, ehara i te mea i tuhia noa.

Te whakamāori i te kore-rārangitanga hei tautuhi i te take

Ina kōpiko te ānau, mā te hanga o te hapa e tohu tōna take. Ko te kōpikopiko whakararo i te ngongotanga tiketike, me te hanga tūāwhiorangi kāore i rerekē, e tohu ana ki te taputapu; i te nuinga o te wā, nā te rama kotiti e whakaiti hē ana i ngā ngongotanga kaha. Ko te kōpikopiko me te pikinga o te pakihiwi 610 nm e tohu ana ki te matū: he rerekē te ngongo a te dimere me ngā huinga nui ake i tō te monomere. Ko tētahi tauira pātea kāore e tae ki te kore, ko ngā tauira paerewa rānei e heke ana i te haere o te wā, e tohu ana ki te ipu: te piri ki ngā mata karāhe, ki ngā ipu ine rānei, otirā ki te rerekē ngā wā pā puta noa i te raupapa ine. Ā, ko tētahi rārangi pai mā ngā ira, engari e whakaputa tonu ana i ngā hua tauira hē, e tohu ana ki te taiao matū: he rerekē ngā tauira i ngā tauira whakatikatika, nō reira hangaia anō te ānau ki roto i te taiao matū o te tauira, whakawhiti rānei ki te tāpiri paerewa. I ia āhuatanga, whakaahuatia ngā toenga ki te kukū. Mā te marara matapōkere e tautoko te tauira pāngarau rārangi tōtika; mā te hanga ataata, mā te hanga kōpiko whakararo rānei e whakahē pono ake i tētahi paepae R pūrua, he kōrero e tautokona māriretia ana e ngā aratohu whakamana.

Ka whai hua tēnei āta whakahaere mō te kikorangi metilīni. He kaha tōna pae monomere, kua tino rangahaua ōna āhuatanga matū, ā, ka kitea ngā tohu o ōna hapa i te tūāwhiorangi i mua i te whakakino i ngā tau. Hangaia te ānau ki tētahi awhe whāiti, whakamahia te tauira pātea pono, mahia ngā tāruarua motuhake, ā, waiho mā te hanga o ngā tūāwhiorangi e whakatau te awhe. He poto te rārangi ka puta, engari he pono ia ira kei runga.

E whakamahi ana ngā tātai whakamārama i tēnei tuhinga i ngā tau tito hei whakaatu i te tikanga. Kei te tohutoro tūāwhiorangi kua honoa ngā whakarea tohutoro i inea me ō rātou āhuatanga katoa, ā, kua whakarārangihia te rangahau whakahuihui taketake ki PMID 33251415.