脳血管障害後の早期運動療法介入が脳の損傷と回復に与える影響について

2011-2012年度科学研究費助成事業(科学研究費補助金(若手研究(B))研究成果報告書　課題番号：23700609　研究代表者：松田史代(鹿児島大学・医学部・助教)本研究では、脳血管障害後、早期からの運動療法介入（本研究ではトレッドミル運動）が梗塞巣や梗塞周辺部（通称ペナンブラ領域）、身体活動全体に及ぼす影響を検討することを目的とした。脳梗塞は、一般的に広く国際的に使われている塞栓糸を使用した左中大脳動脈閉塞・再開通モデルを使用した。作成翌日に、無作為にトレッドミル運動群・非運動群と分け、運動群は毎日20分間トレッドミル運動を行った。また、術後毎日運動機能評価と神経学的評価を実施した。運動開始一週間後より運動群の方が非運動群よりも身体機能の改善が良好にみられ、28日後有意に改善した。梗塞巣周辺部でも運動群の方が非運動群に比べて有意に神経栄養因子ミッドカイン増加した。そのため、ミッドカインと神経修復の関与を検討するために、ミッドカインのノックアウトマウスを使用し、脳梗塞（開頭し末梢中大脳動脈を電気メスで凝固閉塞した）を作成したところ、ミッドカインノックアウトマウスは野生型マウスと比較して、脳梗塞巣体積に関しては、有意差がみられなかったが、反応生アストロサイトや活性型ミクログリアの発現が少ない傾向にあった。前述したラットの梗塞作成後運動を行った群では有意にミッドカインの発現が早期に増加しており、梗塞周辺部の修復にミッドカインの関与が示唆された。しかし、ミッドカインの発現機序や発現作用は、未だ不明な点が多く、今後ミッドカインノックアウトマウスを使用しての更なる解明が必要である。また、何故早期からの運動介入により神経栄養因子が増加するのか、また他の神経栄養因子との関与も含めて更なる検討が必要であ る。The purpose of this study was investigate that the effect of early exercise (this study used the treadmill exercise) after cerebral infarction in the rat. Next day after surgery, stroke rats were randomly assigned to two groups: treadmill-exercised group or non-exercised controls. After ischemia, the rats were received to run on a treadmill for 20 min once a day. All rats were observed the absence of motor coordination and neurological deficits after surgery. Over time motor coordination and neurological deficits improved more in the exercised group than in the non-exercised group. The cellular expression levels of MK were significantly increased in the peri-infarct area of the exercised rats. Therefore The purpose is to confirm the effect of MK gene knockout on cerebral infarction in mouse and the relation of MK with the regeneration. The ratios of infarct volume to cerebral hemisphere were not different between KO and wild type mice at 3 days and 7days. The expression of glial fibrially acidic protein (GFAP) was more and more expressed in periinfarct area after insult. GFAP staining areas in astrocytes (calculated as a ratio of stained areas to hemisphere)were significantly decreased in parietal lobe of KO mice compared with that in wild type mice at 3 days, however, it was not significantly different at 7 days. Since MK has a role as a chemotaxis of microglias and macrophages, the Iba1 expression was studied with immunostaining, which showed no significant difference. The results were suggestive that in KO mice the expression of GFAP was delayed compared with in wild type mice. MK participates in the development of the reactive astrocytes after cerebral infarction

脳血管障害後の早期運動療法介入が脳の可塑性に与える影響について

2009-2010年度科学研究費補助金(若手研究(B))研究成果報告書 課題番号：21700539　研究代表者：松田史代 (鹿児島大学医学部助教)本研究では、リハビリテーションの視点から、リハビリテーション介入が脳血管障害後の脳の可塑性を促せるのか、脳血管障害後の神経栄養因子MKの働きについて検討するとともに、神経栄養因子が神経細胞死や神経修復にどのように働いているのか検討することを目的とする。経時的に脳梗塞モデルを作成し、術1日後よりトレッドミル運動介入を行い、トレッドミル介入群と非介入群における脳梗塞体積測定後、パラフィン切片作成し、免疫組織化学染色にて観察した。その結果、免疫組織化学染色結果より、家兎MK 免疫組織化学染色にて、トレッドミル介入群が非介入群に比べて3日後で有意に発現面積が増加していた。また、神経学的機能では 運動群が非運動群よりも回復が早く、運動機能（棒上歩行機能評価）でも運動群で非運動群と比較し、良好な運動機能回復がみられた。The purpose of this study was to investigate the effects of early treadmill training after cerebral infarction in rats. This study might provide a basic evidence of rehabilitation medicine. Fifty-four- 8-week old male Wistar rats weighing 200-300g were used in this study. The filament (4-0 nylon suture with blunted tip) was inserted into the left common carotid artery by an arteriotomy. It was then passed up through the lumen of the internal carotid artery and lodged in the narrow proximal anterior cerebral artery, blocking the MCA at its origin. Reperfusion was established by withdrawal of the filament by 90 min. Next day after surgery, stroke rats were randomly assigned to two groups: treadmill-exercised group or non-exercised controls. After ischemia, the rats were received to run on a treadmill for 20 min once a day. All rats were observed the absence of motor coordination and neurological deficits after surgery. The improvement of the motor behavioral and the neurological score in the exercised groups was better than those in non-exercised groups. Particularly, the score in the exercised groups was significantly improved compared with those in non-exercised groups at 14 days after surgery (P<0.05). The runner had the increased expression of midkine (MK) over the non-runner with or without MCA occlusion for 3 days

Synthesis of pacidamycin analogues via an Ugi-multicomponent reaction

The second-generation synthesis of 3'-hydroxypacidamycin D (2) has been accomplished via an Ugi-four component reaction at a late stage of the synthesis. This approach provided ready access to a range of analogues including diastereomers of the diaminobutylic acid residue and hybrid-type analogues of mureidomycins. Biological evaluations of these analogues indicated that the stereochemistry at the diaminobutylic acid residue has a crucial impact on both the MraY biochemical inhibition and whole-cell anti-bacterial activity. (C) 2012 Elsevier Ltd. All rights reserved

zds1, a Novel Gene Encoding an Ortholog of Zds1 and Zds2, Controls Sexual Differentiation, Cell Wall Integrity and Cell Morphology in Fission Yeast

While screening for genes that reverse the sporulation-deficient phenotype of the ras1Δ diploid Schizosaccharomyces pombe strain, we identified zds1. This gene shares sequence homology with the ZDS1 and ZDS2 genes from Saccharomyces cerevisiae, which appear to be involved in multiple cellular events. Expression of Zds1 in ras1Δ diploid cells elevated their sporulation rate from 0.3 to 11.2%. Expression of the Zds1 C-terminal region increased the sporulation rate further (to 21.9%) while introduction of the Zds1 N-terminal region had no effect. zds1 expression did not induce sporulation in strains with mutations in genes participating in the downstream MAP kinase cascade. The zds1-disrupted strain is sensitive to CaCl(2), and this effect is suppressed by the C-terminal region of Zds1. The growth of the zds1Δ strain is markedly inhibited by cold temperatures, while its viability decreased in the stationary phase. Moreover, the zds1Δ strain is round in shape and very sensitive to zymolyase, and its cell wall becomes thicker than that of wild type. Thus, zds1 must be required to maintain cell wall integrity. The Zds1–GFP fusion protein localized to the cytosol, the septum, and the cell cortex. Its localization in the septum was dependent on its C-terminal region. Overexpression of the C-terminal region of Zds1 induced multi-septa and abnormal zygotes. We propose that the C-terminal region is the functional domain of Zds1 while the N-terminal region is a negative regulatory region. Thus, Zds1 is involved in multiple cellular events in fission yeast, including sexual differentiation, Ca(2+) tolerance, cell wall integrity, viability in the stationary phase, and cell morphology

A Newly Developed Robot Suit Hybrid Assistive Limb Facilitated Walking Rehabilitation after Spinal Surgery for Thoracic Ossification of the Posterior Longitudinal Ligament: A Case Report

Most patients with thoracic ossification of the posterior longitudinal ligament (OPLL) exhibit delayed recovery of gait dysfunction after spinal injury. The hybrid assistive limb (HAL) is a new robot suit controlling knee and hip joint motion by detecting very weak bioelectric signals on the surface of the skin. This study is to report the feasibility and benefits of patient-assistive HAL walking rehabilitation for facilitating locomotor function after spinal surgery. The patient was a 60-year-old woman with thoracic OPLL, and her motor and sensory paralyses did not improve after spinal surgery, indicating severe impairment in the paretic legs. The subject underwent 6 HAL sessions per week for 8 weeks, consisting of a standing and sitting exercise and walking on the ground with HAL. Clinical outcomes were evaluated before and after HAL training and 1 year after surgery. The subject improved considerably as a result of HAL training. Subsequently, her walking ability recovered rapidly, and she was able to walk unaided six months after surgery. This case study suggests that HAL training is a feasible and effective option to facilitating locomotor function and the early HAL training with physiotherapy may enhance motor recovery of patients with residual paralysis after surgery

Non-adherent cell-specific expression of DOCK2, a member of the human CDM-family proteins

AbstractHuman DOCK180, which was originally identified as a major protein bound to the Crk oncogene product, is an archetype of the CDM family of proteins, including Ced-5 of Caenorhabditis elegans and Mbc of Drosophila melanogaster. After DOCK180, at least three putative human proteins that manifest high amino acid sequence similarity to DOCK180 have been registered in the GenBank/EMBL database. We have designated one of them, KIAA0209, as DOCK2 and characterize here. DOCK2 mRNA was expressed mostly in peripheral blood cells, followed by slight expression in the spleen and thymus, whereas DOCK180 was expressed in all tissues tested except in peripheral blood cells. Immunostaining of human cadaver tissues revealed that the expression of DOCK2 was limited to the lymphocytes and macrophages of various organs. DOCK2 bound to and activated Rac1, as did DOCK180; however, DOCK2 did not bind to CrkII, which transduces signals at focal adhesions. Thus, DOCK180 and DOCK2 are regulators of Rac and function in adherent and non-adherent cells, respectively