１. 脳特異的DISC1結合タンパク質DBZ はオリゴデンドロサイトの分化を促進する発表内容

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Trial for establishment of a new convenient method for quantification of the disease-specific dephosphorylated isoform of an autoimmunogen U1-68K.

publisher[要旨]全身性エリテマトーデス(SLE)や混合性結合組織病(MCTD)などの自己免疫疾患患者の末梢血リンパ球では, 自己抗原U1-small nuclear ribonucleoprotein 68k (U1-68k) のリン酸化が外れた低リン酸化亜型が増加していることが近年発見された. この低リン酸化U1-68kは自己免疫疾患の新たな早期診断マーカーとなる可能性がある. しかしながら, 従来法である細胞の核分画を用いた二次元western blot法は全行程に5日かかることから, より簡便な定量法の開発が必要である. そこで, 本研究ではphos-tag SDS-PAGE後にwestern blotを行う方法(phos-tag western blot法)と, 等電点電気泳動を行った後に直接western blotを行う方法(等電点western blot法)を検討した. その結果, phos-tag western blot法はU1-68kのリン酸化亜型を十分に分離することができず低リン酸化U1-68kの定量には不適であることが示された. それに対し, 等電点western blot法では, pI6.8-9.1の間に10本のU1-68kのバンドが確認され、また, そのうちの1本が目的とする低リン酸化体の等電点に一致した. ここから本方法が従来法に代わる方法として使用できる可能性が示された. 我々は更に, 細胞核画分の代わりに細胞の全抽出液を用いることが可能かを評価した. 5種類の細胞抽出液による全細胞抽出液を比較したところ, SDS-PAGEに通常用いられるSDS処理液が最も効率的にU1-68kを抽出することができ, また二次元western blot法によってU1-68kが複数のスポットとして検出できることを確認した. しかしながら, スポットが観察される領域は大きく酸性側にシフトしており, タンパク質に強く結合したSDSを除去する必要があることが示唆された. [Abstract] Systemic autoimmune diseases such as systemic lupus erythematosus (SLE) or mixed connective tissue diseases (MCTD) are immunologically characterized by generation of various species of autoantibodies in sera from patients. Recently, using 2DE-western blot, we have found that a dephosphorylated form of U1-68k, a major autoanigen to the anti-RNP autoantibody, was increased in peripheral blood lymphocytes from SLE or MCTD patients. This dephosphorylated U1-68k might be used as a new diagnostic marker which could diagnosis the diseases in earlier phase. In this paper, to establish a new convenient method for quantification of the dephosphorylated form, U1-68k isoforms in nuclear fraction of Jurkat cells were separated and detected by two methods, a western blot followed by a phos-tag SDS-PAGE (phos-tag-western blot), and a western blot followed by an isoelectric focusing electrophoresis (IEF-western blot). As a result, phos-tag-western blot showed only two bands of U1-68k isoforms, indicating that this method is inadequate for our purpose. On the other hand, IEF-western blot showed at least 10 bands of U1-68k within pI range of 6.8-9.1, and one of the bands showed the same pI value (7.9) as those of the disease-specific dephosphorylated U1-68k detected by 2DE-western blot. This indicated that the IEF-western blot could be used as a new convenient method for quantification of the dephosphorylated form. In addition, we examined whether U1-68k isoforms in a whole cell extract could be detected by 2DE-westen blot. We tested 5 popular cell lysis solutions, and found that the SDS sample buffer, which is commonly used for SDS-PAGE, extract U1-68k with the highest yield, and showed multiple U1-68k spots by the 2DE-western blot. However, these spots were detected in the acidic pI regions (pI 3-5), indicating that SDS molecules may bound strongly to the U1-68ks and shift their isoelectric points. Therefore, it must be needed to eliminate the strongly combined SDS-molecules before 2DE or IEF-western blot

〈Note〉The Oral Introduction of English Target Sentences in Elementary School Education : An Analysis of the Present Situation and a Suggestion for Developing ICT Support Materials

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〈Articles〉A Cognitive Approach to the Concept of Enter into NP

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publisher<Synopsis>We carried out a biological research at the Yaeyama islands in Okinawa Prefecture, from March 25 to April 1, 2013. The research points were Ishigaki, Iriomote and Taketomi islands. In a total of three islands, 50 species belonging to five families of butterflies were observed. Only 50 species of butterflies were recorded because of the bad and cold weather. We collected migrating butterflies. Especially, six species of the butterflies, Hypolimnas bolina, Symbrenthia lilaea, Euploea Sylvester, Parantica aglea, Parantica swinhoei and Leptosia nina, may have established the population temporally in these islands

<Practical investigations and case studies>Teaching Materials of the Health and Sports Science at Kinki University Correspondence Course (4): Using the Beachball Volleyball

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カイコガ抑制性グルタミン酸受容体の遺伝子発現とケミカルプローブによる制御に関する研究

none学位の種類：農学　　学位授与年月日：平成27年3月23日　　主査：松田, 一彦 教授　　報告番号：甲第1235号　　学内授与番号：農第199

小児IgA腎症における肩桃摘出の効果とその機序に関する検討

none学位の種類：医学　　学位授与年月日：平成 27年3月23日　　主査：竹村, 司 教授　　報告番号：甲第1242号　　学内授与番号：医第1163

〈Original 〉5-Aminolevulinic acid reduces food intake

publisher<Synopsis>5-aminolevulinic acid (ALA) is precursor of tetrapyrrole compounds such as heme, chlorophyll and cobalamin. Recent study has been shown that ALA reduces glucose levels during fasting and after glucose loading in prediabetic subjects. Previously, we reported that administration of ALA ameliorates diabetic conditions such as blood glucose levels and hemoglobin A1c in Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model of type 2 diabetes mellitus. Furthermore, it was indicated that ALA reduces food intake and body weight in OLETF rats. The mechanisms of food intake reduction by ALA is not clear. We demonstrated that the effect of intragastrical and intracerebroventricular administration of ALA and the central GABAergic system of food intake. SD rats were intragastrically or intracerebroventricularly administrated ALA. Food intake reductions was shown in three hours after intragastrical administration of ALA. It was also shown that intracerebroventricular administration of ALA reduces food intake. In this study, the effect of GABAergic system of food intake of ALA was not clear. However, these results suggest that ALA reduces food intake via central nervous systems