ʻO ka ho'āʻo ʻana o Ames ma nā haʻawina Genotoxicity: He Pōhaku kihi o ka hoʻāʻo ʻana i ka ʻawaʻawa ma lalo o ka OECD 471 alakaʻi.

Hua'ōlelo:Ho'āʻo Ames, hoʻāʻo Ames liʻiliʻi, hoʻāʻo ʻo Ames Microfluctuations, hoʻāʻo Fluctuations, Ames II, Induced S9, S9 Mix, Pūnaehana hoʻōla Metabolic, Enhanced Ames Test, Bacterial Reverse Mutation Test, OECD 471, Mutagenicity test, Genotoxicity, Genetic Toxicity

Nā huahana IPHASE

Introduction to the Ames Test and its role in Genetic Toxicity

ʻO kaHoao Ames, i ʻike ʻia ʻo kaʻO ka hoʻāʻo ʻana i ka hoʻololi ʻana o ka bacteria, he kumu hoʻāʻo make kinoahoʻāʻo mutagenicity. Hoʻomohala ʻia e Kauka Bruce Ames i ka makahiki 1970, ʻike ka hoʻāʻo i ka hikiʻona ʻonama ka loiloi ʻana i ka hiki o nā mea kemika e hoʻoulu i nā hoʻololi i ka DNA o ka bacteria. Ua lilo kēia ho'āʻo i mea hoʻoponopono ma ka honua holoʻokoʻa a ʻike ʻia ma lalo o ke alakaʻiOECD 471, ka mea e hāʻawi i ke kāʻei no ka hoʻokō ʻana i ka hoʻāʻo me ka hilinaʻi a me ka reproducibly.

Ke Kumu o ka Ames Test ma Genotoxicity Studies

Ma kona kumu, loiloi ka Ames Test i ka hiki o kekahi mea ke hoʻoulu i nā hoʻololi ʻana i nā ʻano ʻano ʻenehana kūikawā o Salmonella typhimurium a i ʻole Escherichia coli i hemahema i ka synthesis histidine. ʻAʻole hiki ke ulu kēia mau maʻi bacteria i loko o ka histidine-deficient media ke ʻole kahi hoʻololi e hoʻihoʻi i ko lākou hiki biosynthetic. Ke loaʻa kahi mutagen, hoʻonui ia i ka nui o ia mea“hoʻololi hoʻololi,”e hōʻike ana i ke ʻano o ka genetic toxicity.

Hāʻawi kēia assay i kahi ala maʻalahi akā ikaika e wānana i ka mutagenicity a me ka carcinogenicity hiki i ke kanaka. No laila, ʻo ia kahi mea hana hoʻomaʻamaʻa mua i nā haʻawina genotoxicity no nā kemika, nā lāʻau lapaʻau, nā mea hoʻohui meaʻai, a me nā laʻana kaiapuni.

Ka Hana o ka Metabolic Activation: Induced S9 and S9 Mix

ʻO kahi hiʻohiʻona koʻikoʻi o kekahi hoʻokolohua mutagenicity, me ka Ames Test, ʻo ia ka hoʻokomo ʻana o aʻōnaehana hoʻoikaika metabolic. ʻAʻole mutagenic pololei ka nui o nā kemika akā koi i ka hoʻololi metabolic e hōʻike i nā hopena genotoxic. ʻO kēia kahi o kaS9 Hui—he hoʻomākaukau ʻana i nā enzymes o ke ake, i loaʻa maʻamau maihoʻokomo ʻia ʻo S9nā hakina o nā ʻauʻau rodent—hele mai i ka pāʻani. Hoʻohālikelike ka S9 Mix i ka metabolism mammalian, e hāʻawi ana i kahi loiloi pili physiologically o ka pūhui hoʻāʻo's hiki mutagenic.

Hoʻohana pinepine ʻia nā mea hana induction e like me Aroclor 1254 e hoʻonui i ka ʻike enzyme o nā ʻiʻo ate i hoʻohana ʻia no ka hoʻomākaukau ʻana i ka hapa S9. ʻO ka hoʻokomo ʻia ʻana o kēia ʻōnaehana S9 i hoʻokomo ʻia e hōʻoia i ka hoʻololi ʻia ʻana o nā promutagens (nā puʻupuʻu e koi ana i ka hoʻōla) i kā lākou mau ʻano mutagenic ikaika i ka wā o ka hoʻāʻo ʻana.

Ho'āʻo Mini Ames a me ka ho'āʻo ʻana o Ames Microfluctuations: Nā ʻano ʻokoʻa paʻa a maikaʻi

ʻO nā holomua hou i alakaʻi i ka hoʻomohala ʻana i nā ʻano miniaturized e like me kamini Ames hoaoaAmes Microfluctuations ho'āʻo, i ʻike ʻia ʻo kaHo'āʻo Fluctuations. Hoʻemi nui kēia mau ʻano i ka nui o nā mea hoʻāʻo a me nā reagents e pono ai, e kūpono ai no ka nānā mua ʻana a me nā kaiapuni kiʻekiʻe.

Hoʻololi ka Fluctuations Test i nā pā agar me nā moʻomeheu wai i nā pā microtiter a ana i nā loli pH ma ke ʻano he hōʻailona o ka ulu ʻana o ka maʻi bacteria, e hiki ai ke ʻike wikiwiki, kumu kūʻai pono o ka hana mutagenic. Mālama kēia mau ʻano hana hou i ke koʻikoʻi ʻepekema o ka hoʻomaʻamaʻa kuʻuna me ka hoʻomaikaʻi ʻana i ka pono o ka hana, ʻoi aku hoʻi i ka nānā ʻana i nā hale waihona puke nui.

Ames II: Hoʻonuiʻia kaʻike i ka ho'āʻo Mutagenicity

ʻO kaAmes IIHōʻike ka hoʻāʻo i kahi loli hou i ka hoʻāʻo mutagenicity, i hoʻolālā ʻia e hoʻonui i ka ʻike ʻike no nā mutagens nāwaliwali. Hoʻohui ia i ka ʻōnaehana hoʻāʻo Ames maʻamau me nā moʻomeheu wai hoʻokuʻu ʻia, e ʻae ai i ke kiʻekiʻe o ka cell density a me nā manawa hōʻike lōʻihi. Hoʻonui kēia hoʻonohonoho i ka hiki ke hopu i nā hopena mutagenic maʻalahi, e hoʻomaikaʻi i ka pololei o ka loiloi pilikia, ʻoi aku hoʻi no ka palena palena a i ʻole nā ​​genotoxins haʻahaʻa.

Ho'āʻo Ames hoʻonui a me ka hoʻāʻo ʻana i ka hoʻololi ʻana o ka bacterial ma lalo o OECD 471

ʻO kaHo'āʻo Ames i hoʻonui ʻia, i kekahi manawa i kapa ʻia ʻo Bacterial Reverse Mutation Test, e pili ana me nā alakaʻi honua ma lalo o OECD 471. Hōʻike kēia alakaʻi i ke ʻano maʻamau no ka hana ʻana i ka Ames assay, me ke koho ʻana i nā kānana, ka hoʻohana ʻana i ka metabolic activation, ka wehewehe ʻana i ka ʻikepili, a me ka helu helu helu.

ʻO ka hoʻokō ʻana me OECD 471 e hōʻoia i ka hilinaʻi a me ka hana hou ʻana o nā hopena hoʻāʻo, e ʻae iā lākou i nā kino hoʻoponopono e like me FDA, EMA, a me EPA. Hoʻokomo pinepine ka Enhanced Ames Test i nā kānana he nui, me ka ʻole o ka S9 metabolic activation system, e hāʻawi i kahi loiloi mutagenicity holoʻokoʻa.

Ka hopena: ʻO ka waiwai mau loa o ka Ames Test i ka Genetic Toxicity Evaluation

Ma ke ʻano o ka hoʻāʻo ʻana i ka genetic toxicity, ʻo ka Ames Test a me kāna mau derivatives hou e noho pono ʻole. Inā hana ʻia e like me ka hoʻohui ʻana o ka pā papa kahiko a i ʻole ka mini Ames, Ames II, a i ʻole Fluctuations Test, hāʻawi mau ia i ka ʻikepili hilinaʻi e kākoʻo i nā loiloi palekana. Ma ka hoʻohui ʻana i ka metabolic activation ma o S9 Mix, ma muli o nā kūlana OECD 471, a me ka hoʻonui ʻana i ka ʻenehana, hoʻomau ka Ames Test i nā pono o ka noiʻi genotoxicity o kēia manawa.