The Vesicle Priming Factor CAPS Functions as a Homodimer via C2 Domain Interactions to Promote Regulated Vesicle Exocytosis.

Neurotransmitters and peptide hormones are secreted by regulated vesicle exocytosis. CAPS (also known as CADPS) is a 145-kDa cytosolic and peripheral membrane protein required for vesicle docking and priming steps that precede Ca2+-triggered vesicle exocytosis. CAPS binds phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and SNARE proteins and is proposed to promote SNARE protein complex assembly for vesicle docking and priming. We characterized purified soluble CAPS as mainly monomer in equilibrium with small amounts of dimer. However, the active form of CAPS bound to PC12 cell membranes or to liposomes containing PI(4,5)P2 and Q-SNARE proteins was mainly dimer. CAPS dimer formation required its C2 domain based on mutation or deletion studies. Moreover, C2 domain mutations or deletions resulted in a loss of CAPS function in regulated vesicle exocytosis, indicating that dimerization is essential for CAPS function. Comparison of the CAPS C2 domain to a structurally defined Munc13-1 C2A domain dimer revealed conserved residues involved in CAPS dimerization. We conclude that CAPS functions as a C2 domain-mediated dimer in regulated vesicle exocytosis. The unique tandem C2-PH domain of CAPS may serve as a PI(4,5)P2-triggered switch for dimerization. CAPS dimerization may be coupled to oligomeric SNARE complex assembly for vesicle docking and priming

Unusual structures are present in DNA fragments containing super-long Huntingtin CAG repeats.

BACKGROUND: In the R6/2 mouse model of Huntington's disease (HD), expansion of the CAG trinucleotide repeat length beyond about 300 repeats induces a novel phenotype associated with a reduction in transcription of the transgene. METHODOLOGY/PRINCIPAL FINDINGS: We analysed the structure of polymerase chain reaction (PCR)-generated DNA containing up to 585 CAG repeats using atomic force microscopy (AFM). As the number of CAG repeats increased, an increasing proportion of the DNA molecules exhibited unusual structural features, including convolutions and multiple protrusions. At least some of these features are hairpin loops, as judged by cross-sectional analysis and sensitivity to cleavage by mung bean nuclease. Single-molecule force measurements showed that the convoluted DNA was very resistant to untangling. In vitro replication by PCR was markedly reduced, and TseI restriction enzyme digestion was also hindered by the abnormal DNA structures. However, significantly, the DNA gained sensitivity to cleavage by the Type III restriction-modification enzyme, EcoP15I. CONCLUSIONS/SIGNIFICANCE: "Super-long" CAG repeats are found in a number of neurological diseases and may also appear through CAG repeat instability. We suggest that unusual DNA structures associated with super-long CAG repeats decrease transcriptional efficiency in vitro. We also raise the possibility that if these structures occur in vivo, they may play a role in the aetiology of CAG repeat diseases such as HD

18FDG-PET at 1-Month Intervals Is a Better Predictive Marker for GISTs That Are Difficult to Be Diagnosed Histopathologically: A Case Report

Imatinib mesylate is a tyrosine kinase inhibitor of c-KIT and PDGFRA. Imatinib mesylate is an effective drug that can be used as a first-choice agent for treatment of GISTs. Prior to treatment, molecular diagnosis of c-KIT or PDGFRA is necessary; however, in some types of GISTs, it is impossible to obtain a sufficient amount of specimen for diagnosis. An inoperable or marginally resectable GIST in a 79-year-old female was difficult to be diagnosed at a molecular pathological level, and hence, exploratory treatment was initiated using imatinib combined with 18FDG-PET evaluation at 1-month intervals. PET imaging indicated a positive response, and so we continued imatinib treatment in an NAC setting for 4 months. As a result, curative resection of the entire tumor was successfully performed with organ preservation and minimally invasive surgery. 18FDG-PET evaluation at 1-month intervals is beneficial for GISTs that are difficult to be diagnosed histopathologically

Ligilactobacillus salivarius strains isolated rrom the porcine gut modulate innate immune responses in epithelial cells and improve protection against intestinal viral-bacterial superinfection

Previously, we constructed a library of Ligilactobacillus salivarius strains from the intestine of wakame-fed pigs and reported a strain-dependent capacity to modulate IFN-β expression in porcine intestinal epithelial (PIE) cells. In this work, we further characterized the immunomodulatory activities of L. salivarius strains from wakame-fed pigs by evaluating their ability to modulate TLR3- and TLR4-mediated innate immune responses in PIE cells. Two strains with a remarkable immunomodulatory potential were selected: L. salivarius FFIG35 and FFIG58. Both strains improved IFN-β, IFN-λ and antiviral factors expression in PIE cells after TLR3 activation, which correlated with an enhanced resistance to rotavirus infection. Moreover, a model of enterotoxigenic E. coli (ETEC)/rotavirus superinfection in PIE cells was developed. Cells were more susceptible to rotavirus infection when the challenge occurred in conjunction with ETEC compared to the virus alone. However, L. salivarius FFIG35 and FFIG58 maintained their ability to enhance IFN-β, IFN-λ and antiviral factors expression in PIE cells, and to reduce rotavirus replication in the context of superinfection. We also demonstrated that FFIG35 and FFIG58 strains regulated the immune response of PIE cells to rotavirus challenge or ETEC/rotavirus superinfection through the modulation of negative regulators of the TLR signaling pathway. In vivo studies performed in mice models confirmed the ability of L. salivarius FFIG58 to beneficially modulate the innate immune response and protect against ETEC infection. The results of this work contribute to the understanding of beneficial lactobacilli interactions with epithelial cells and allow us to hypothesize that the FFIG35 or FFIG58 strains could be used for the development of highly efficient functional feed to improve immune health status and reduce the severity of intestinal infections and superinfections in weaned piglets.Fil: Indo, Yuhki. Tohoku University; JapónFil: Kitahara, Shugo. Tohoku University; JapónFil: Tomokiyo, Mikado. Tohoku University; JapónFil: Araki, Shota. Tohoku University; JapónFil: Islam, Md Aminul. Tohoku University; Japón. Bangladesh Agricultural University; BangladeshFil: Zhou, Binghui. Tohoku University; JapónFil: Albarracín, Leonardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Exactas y Tecnología; ArgentinaFil: Miyazaki, Ayako. National Institute of Animal Health; JapónFil: Ikeda Ohtsubo, Wakako. Tohoku University; JapónFil: Nochi, Tomonori. Tohoku University; JapónFil: Takenouchi, Takato. National Agriculture And Food Research Organization; JapónFil: Uenishi, Hirohide. National Agriculture and Food Research Organization; JapónFil: Aso, Hisashi. Tohoku University; JapónFil: Takahashi, Hideki. Tohoku University; JapónFil: Kurata, Shoichiro. Tohoku University; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Kitazawa, Haruki. Tohoku University; Japó

Risk-adjusted therapy for pediatric non-T cell ALL improves outcomes for standard risk patients: results of JACLS ALL-02

This study was a second multicenter trial on childhood ALL by the Japan Childhood Leukemia Study Group (JACLS) to improve outcomes in non-T ALL. Between April 2002 and March 2008, 1138 children with non-T ALL were enrolled in the JACLS ALL-02 trial. Patients were stratified into three groups using age, white blood cell count, unfavorable genetic abnormalities, and treatment response: standard risk (SR), high risk (HR), and extremely high risk (ER). Prophylactic cranial radiation therapy (PCRT) was abolished except for CNS leukemia. Four-year event-free survival (4yr-EFS) and 4-year overall survival (4yr-OS) rates for all patients were 85.4% ± 1.1% and 91.2% ± 0.9%, respectively. Risk-adjusted therapy resulted in 4yr-EFS rates of 90.4% ± 1.4% for SR, 84.9% ± 1.6% for HR, and 66.5% ± 4.0% for ER. Based on NCI risk classification, 4yr-EFS rates were 88.2% in NCI-SR and 76.4% in NCI-HR patients, respectively. Compared to previous trial ALL-97, 4yr-EFS of NCI-SR patients was significantly improved (88.2% vs 81.2%, log rank p = 0.0004). The 4-year cumulative incidence of isolated (0.9%) and total (1.5%) CNS relapse were significantly lower than those reported previously. In conclusion, improved EFS in NCI-SR patients and abolish of PCRT was achieved in ALL-02

ニンチショウ カンゴ ニンテイ カンゴシ キョウイク カテイ ニオケル キョウイク ト ジュギョウ ヒョウカ ノ ブンセキ

認定看護師教育課程 認知症看護認定看護師コース(以下、本教育課程)の教育実践を振り返り、今後の課題を検討した。今回、授業評価を実施した研修生20名の認知症看護経験年数は平均10.4±4.45年であり、本教育課程を受講した理由の大半は自己の学習ニーズ(16名)であった。研修生の本教育課程に対する期待には「臨床実践に沿った講義内容」があり、修了生による臨床に即した実践内容を含んだ授業の工夫が必要であると考えられた。研修生は「認知症ケアを自己の言語で伝えられるようになりたい」という目標を持つ者が多く、本教育課程で看護過程の展開技術を重視した学習プログラムはニーズに一致していた。研修生は自己の目標達成において病院実習の有効性を高く評価していたが、満足度は低い傾向を示した。特に、受講理由が施設長等の勧めである研修生にその傾向が認められた。よって、入学前に研修生が自己を振り返る課題を課し、本教育課程での学びのイメージを高めさせ、認定看護師の研修の質の向上を図ることが課題に挙げられた。The purpose of the research was to look back upon the educational practice of the course for certified nurses in dementia nursing of the curriculum for of certified nurse education, (hereafter referred as to "this education course") and discuss tasks for future use. The average number of years of experience in dementia nursing for the 20 trainees of whom the class evaluation of this education course was implemented was 10.4±4.45, and the reason for attending this educational course was for the majority, a fulfillment of their own learning requirements (16 trainees). Their expectations for this educational course included the "contents of lectures consistent with clinical practice." Therefore, it was considered necessary to contrive the class in line with the clinical practice of those who have already completed this educational course. There were many trainees who had the target of "conveying dementia care in their own words," which showed that a learning program with emphasis on developing technology for the nursing process matched their needs. Although the trainees highly evaluated the eff ectiveness of hospital practice for their target achievement, they showed a low degree of satisfaction. This trend was significant among those who attended this educational course at the encouragement of the chief of their facility, in particular. Therefore, the benefit of indicating the tasks before they attend this educational course, in order to make them refl ect on themselves, enhance their image of learning in this educational course. Improving the quality of training for qualified nurses was also clarified

原子間力顕微鏡を用いた生体分子の力学測定とマッピング : 原子間力顕微鏡プローブ修飾法の開発と応用

京都大学0048新制・課程博士博士(生命科学)甲第16232号生博第219号新制||生||29(附属図書館)28811京都大学大学院生命科学研究科統合生命科学専攻(主査)教授 竹安 邦夫, 教授 井上 丹, 教授 小堤 保則学位規則第4条第1項該当Ph.D. (Life Sciences)Kyoto UniversityDA

Research data supporting “The vesicle priming factor CAPS functions as a homodimer via C2 domain interactions to promote regulated vesicle exocytosis”

AFM images of CAPS, both dried onto mica and associated with lipid bilayers.Royal Society [NF110528], NIH [DK040428 and GM119158