{"created":"2023-07-27T07:01:03.667150+00:00","id":61204,"links":{},"metadata":{"_buckets":{"deposit":"97f161d6-5bf9-4ce5-b67e-8da0b2ec3d98"},"_deposit":{"created_by":18,"id":"61204","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"61204"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00061204","sets":["2812:2813:2841"]},"author_link":["20336","71203"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"1994-03-23","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"1991 – 1992","bibliographic_titles":[{"bibliographic_title":"平成4(1992)年度 科学研究費補助金 一般研究(C) 研究成果報告書概要"},{"bibliographic_title":"1992 Fiscal Year Final Research Report Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Nakamura, Shinichi"}],"nameIdentifiers":[{"nameIdentifier":"20336","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"90019620","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=90019620"},{"nameIdentifier":"90019620","nameIdentifierScheme":"研究者番号","nameIdentifierURI":"https://nrid.nii.ac.jp/nrid/1000090019620"}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Clostridium difficile VPI 10463株の超音波破砕菌体抽出液より、ウシサイログロブリンアフィニティクロマトグラフィー(TGAC)モノQカラムクロマトグラフィー(モノQ-FPLC)により高度精製菌体内トキシンAを得るこてが出来た。菌体内トキシンAのTGAC溶出パターンは菌体外トキシンAの場合(HA活性は温度溶出画分に陽性、素通り画分では陰性)と全く異り、HA活性は温度溶出画分では極めて弱く(2^0)、素通り画分では高かった(2^9)。温度溶出画分を透析後(HA活性が2^5に上昇)、モノQ-FPLCを行った。最初のモノQ-FPLCにおいて細胞毒性のピークとHA活性のピークの解離が認められた。高細胞毒性を示した画分について再度モノQ-FPLCを行った結果、細胞毒性のピークのみが認められた。最終標品は未変性PAGE上分子量580kDaの単一バンドとして泳動された。HA活性陰性菌体内トキシンAの細胞毒性、マウス到死活性、結紮腸管液体貯留活性に対する最小有効量は、各々0.83ng、8.7ng、5μgであり、菌体外トキシンAとほぼ等しかった。菌体内・外トキシンAに対する抗体を作製し両トキシンAを血清学的に比較検討した。抗菌体内・外トキシンA抗体を用いたオクタロニーゲル内沈降反応においては、菌体内トキシンAおよび菌体外トキシンAとの間の沈降線は、1本のみ形成され、かつ完全に融合した。細胞毒性、マウス到死活性、結紮腸管液体貯留活性中和試験においては、両抗トキシンA抗体は、対応するトキシンAに対する中和能と同程度の中和能(1:128〜1:256)を、互いに異るトキシンAに対し示した。菌体外トキシンAのHA活性に対しては両抗体共に1:16の中和能を示した。以上の結果は、トキシンAは赤血球に対する結合部位の発現が少い形で菌体内で産生され、菌体外に放出される時、あるいは培養液中で結合部位が完全に発現される可能性を示している。","subitem_description_type":"Abstract"},{"subitem_description":"After sonic disintegration of Clostridium difficile cells, intracellular toxin A was purified to homogeneity by thyroglobulin affinity chromatography (TGAC) followed by successive anion- exchange chromatography on Mono Q incorporated into fast protein liquid chromatography (FPLC) apparatus. High hemagglutination (HA) activity was detected in TGAC-unbound fractions (2^9), but not in TGAC thermal eluates (2^0). The low HA titer of the thermal eluates was markedly increased to 2^5 after dialysis against 0.02 M Tris-HCl (pH 7.5). A disparity in the position of the peak between cytotoxicity and HA activity was observed in the first Mono Q-FPLC. Intracellular toxin A without HA activity was obtained by second Mono Q-FPLC. The molecular weight of the intracellular toxin A was estimated by polyacrylamide gel electrophoresis to be 580 kDa in non-denaturing condition. The minimum doses of the toxin causing cytotoxicity, mouse lethality and enterotoxicity were 0.83 ng, 8.7 ng and 5 ug, respectively. Intracelluler toxin A and extracellular toxin A were compared serologically. In Ouchterlony test, anti-intracellular toxin A and anti-extracellular toxin A sera showed identity of the intracellular and extracellular toxin A. In neutralization tests, both anti-intracellular and anti-extracellular toxin A sera alternatively neutralized the heterologous toxins for cytotoxicity, mouse lethality and loop response at nearly the same titers (1:128 - 1:256) as did the homologous toxins. Although intracellular toxin A lacked HA activity, anti-intracellular toxin A neutralized HA activity of the extracellular toxin A at the same titer (1:16) as anti-extracellular toxin A serum. These findings suggest a possibility that toxin A is synthesized as HA-negative form in cells, which has a few binding sites to the erythrocytes, and converted to HA-positive form,when it is released from cells, or after release from cells.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:03670208, 研究期間(年度):1991 – 1992","subitem_description_type":"Other"},{"subitem_description":"出典：研究課題「Clostridium difficile菌体内トキシンAの精製とその性状の解析」課題番号03670208\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） \n（https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03670208/036702081992kenkyu_seika_hokoku_gaiyo/）を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学医学部","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00067448","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=90019620"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=90019620","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-03670208/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-03670208/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03670208/036702081992kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03670208/036702081992kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"中村, 信一"}],"nameIdentifiers":[{"nameIdentifier":"71203","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"90019620","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=90019620"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2022-10-24"}],"displaytype":"detail","filename":"ME-PR-NAKAMURA-S-kaken 1994-2p.pdf","filesize":[{"value":"92.6 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-NAKAMURA-S-kaken 1994-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/61204/files/ME-PR-NAKAMURA-S-kaken 1994-2p.pdf"},"version_id":"26697d29-f972-41a0-b007-e712bee3bb14"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"Clostridium difficile菌体内トキシンAの精製とその性状の解析","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Clostridium difficile菌体内トキシンAの精製とその性状の解析"},{"subitem_title":"Purification and characterization of intracellular toxin A of Clostridium difficile","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2841"],"pubdate":{"attribute_name":"公開日","attribute_value":"2022-10-24"},"publish_date":"2022-10-24","publish_status":"0","recid":"61204","relation_version_is_last":true,"title":["Clostridium difficile菌体内トキシンAの精製とその性状の解析"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2024-07-01T06:46:09.984183+00:00"}