ヒト糸球体メサンギウム細胞特異的遺伝子のクロ-ニング

近年、腎疾患に対する有効な治療法を確立するため、分子レベルでの病態の解明が望まれている。ヒト糸球体メサンギウム細胞(HMC)は、腎糸球体の生理的に重要な細胞であり、慢性糸球体腎炎等のの増殖性糸球体腎炎の発症において重要な病態生理学的意義を有する細胞である。これまでHMCのcell biologyを明らかにすべく多くの研究が行われてきたが、依然不明の点も多く、有効なマーカータンパクさえも同定されていない。そこで私たちは、 1)HMCに発現している遺伝子を解析し、HMCを構成する遺伝子群の特徴を明らかにすること、2)HMC特異的遺伝子をクローニングし、その構造と機能について解析すること、を研究の目的として研究を開始した。 1:HMCcDNAライブラリーの作製及び解析正常ヒト腎組織より培養メサンギウム細胞を確立した。mRNAを抽出し、HMCに発現する遺伝子解析用の3′-directed cDNAライブラリーを作製した。また、full length cDNAライブラリーも合わせて作製した。3′-directed cDNAライブラリーより任意に1200クロ-ンを抽出しその塩基配列を決定した。その結果、蛋白合成に関与する遺伝子及び分泌蛋白の遺伝子が数多く検出された。最も発現量の多い遺伝子はファイブロネクチンであった。 2:HMC特異的遺伝子の同定上記で得られた解析結果を遺伝子データーバンク或いは他の組織(肝、肺、線維芽細胞、血管内皮細胞等)での解析結果(大阪大学細胞工学センター 大久保助教授より提供)と比較し、10個のHMC特異的且つ未知の遺伝子を選別した。これらのクローンのfull length cDNAをクローニングし解析したところ、クローンNo.9422はセリンプロテアーゼインヒビターファミリーと高い相同性を有する遺伝子であることが明らかになった。科学研究費補助金 研究種目:一般研究(B)(2) 課題番号:07457240 研究代表者:宮田 敏男 研究期間:1995-1996年度research repor

Observation of the ηc(2S) in Exclusive B→KKSK-π+ Decays

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Von treuen, falschen und einstigen Freunden : deutsch-englische etymologische Paare

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PZQ enantiomers do not activate heterologously expressed TRPV1.

(A) Representative fluorescence traces from HEK293 cells loaded with Fluo-4 AM following the addition of capsaicin (1μM, black) or ±PZQ (100μM, purple), followed by addition of ATP (100μM). (B) Representative fluorescence traces from HEK293 cells transfected with hTRPV1 and loaded with Fluo-4 AM following the addition of capsaicin (1μM), followed by addition of ATP (100μM), in the presence of DMSO (0.1%, black) or capsazepine (10μM, brown). (C) Representative fluorescence traces of HEK293 cells transfected with hTRPV1 and loaded with Fluo-4 AM following the addition of ±PZQ (100μM), followed by ATP (100μM), in the presence of DMSO (0.1%, black), or capsazepine (10μM, brown). (D) Cumulative measurements of peak fluorescence ratio (F/F0, where ‘F’ represents fluorescence at peak and ‘F0’ represents fluorescence at time = 0) from Ca2+ imaging experiments under indicated conditions. Data represent population means±s.e.m. (>20 cells) from n≥3 independent transfections.</p

PZQ enantiomers activate TRPA1.

(A) Representative fluorescence traces from HEK293 cells loaded with Fluo-4 AM following the addition of AITC (100μM, black) or ±PZQ (100μM, purple), followed by addition of ATP (100μM). (B) Representative fluorescence traces from HEK293 cells transfected with hTRPA1 and loaded with Fluo-4 AM following the addition of AITC (100μM), followed by addition of ATP (100μM), in the presence of DMSO (0.1%, black) or AM-0902 (1μM, brown). (C) Representative fluorescence traces of HEK293 cells transfected with hTRPA1 and loaded with Fluo-4 AM following the addition of ±PZQ (100μM), followed by ATP (100μM), in the presence of DMSO (0.1%, black), or AM-0902 (1μM, brown). (D) Representative fluorescence traces from HEK293 cells transfected with hTRPA1 and loaded with Fluo-4 AM following the addition of R-PZQ (100μM, red) or S-PZQ (100μM, blue), followed by addition of ATP (100μM). (E) Cumulative measurements of peak fluorescence ratio (F/F0, where ‘F’ represents fluorescence at peak and ‘F0’ represents fluorescence at time = 0) from Ca2+ imaging experiments under indicated conditions. Data represent representing population mean±s.e.m. (>20 cells) from n≥3.</p

Interpretation of the Anomalous Transient Behavior in a Heter-ogeneously Catalyzed Reaction Caused by a Dual Path Way

application/pdfThe complicated transient behavior observed in the oxidation of CO over two differently prepared zinc oxides (K25-ZnO and Kan-ZnO)has been interpreted based on the dual path models.　The characteristic transient response curves are classified into four types : (1) monotonous，(2)overshoot，13)false start and (4)complex types. The mode is sensitively influenced by the magnitude of CO concentration jump given at the inlet of the reactor. The drastic shift between any two characteristic modes can be visualized as a function of COconcentration by using a computer simulation technique.departmental bulletin pape

(<i>S</i>)-PZQ decrease tone of contracted mesenteric arteries.

(A) Representative tension recordings showing addition of ±PZQ (left), (R)-PZQ (50μM, middle) and (S)-PZQ (50μM, right) during a KPSS-evoked contraction. (B) Individual data points (squares) show magnitude of tension change measured at a fixed time interval (1 min) after addition of (R)-PZQ (red) or (S)-PZQ (blue) relative to control traces (DMSO, vehicle) during a KPSS-evoked contraction. Each data point represents a single measurement from a unique vessel (n≥6 measurements for each condition). Average of all measurements represented by the bar chart.</p

Comparison of responses to TRPM8 ligands in wild type and TRPM8 KO mice.

<p>(<b>A-C</b>) Responses to the TRPM8 agonists (A) menthol (300μM), (B) icilin (50μM) and (C) WS-12 (50μM) in wild type (WT, black) and TRPM8 knockout mice (TRPM8 KO, purple) when applied during the sustained phase of a KPSS-evoked contraction (shown by black circle). (<b>D</b>) Similar assay for (<i>S</i>)-PZQ (50μM) evoked relaxation during KPSS-evoked contraction in wild type (WT, black) and TRPM8 knockout mice (TRPM8 KO, purple). (<b>E</b>) Cumulative dataset measuring (<i>S</i>)-PZQ (50μM) evoked relaxation from experiments such as shown in (D). Data represent mean±s.e.m. from averaged measurements from mesenteric vessel strips from n≥3 mice.</p

Heterologously expressed TRPM8 mediates Ca²⁺ influx.

Addition of menthol (300μM, first arrow) to TRPM8-expressing HEK293 cells does not cause a Ca2+ signal in Ca2+-free media, only when extracellular media is replaced with Ca2+-containing media. Traces represent fluorescence profiles from individual cells from a representative experiment. (TIF)</p

Systematic truncation of the hMTPPT polypeptide and their expression pattern in HepG2 cells.

<p>Sub cellular expression of indicated (five) hMTPPT-GFP truncated constructs in HepG2 cells. Lateral (<i>xy</i>) confocal images of different truncated constructs were imaged after 24–48 hrs of transient transfection.</p