Nucleocytoplasmic transport of Alp7/TACC organizes spatiotemporal microtubule formation in fission yeast

Ran GTPase activates several target molecules to induce microtubule formation around the chromosomes and centrosomes. In fission yeast, in which the nuclear envelope does not break down during mitosis, Ran targets the centrosomal transforming acidic coiled-coil (TACC) protein Alp7 for spindle formation. Alp7 accumulates in the nucleus only during mitosis, although its underlying mechanism remains elusive. Here, we investigate the behaviour of Alp7 and its binding partner, Alp14/TOG, throughout the cell cycle. Interestingly, Alp7 enters the nucleus during interphase but is subsequently exported to the cytoplasm by the Exportin-dependent nuclear export machinery. The continuous nuclear export of Alp7 during interphase is essential for maintaining the array-like cytoplasmic microtubule structure. The mitosis-specific nuclear accumulation of Alp7 seems to be under the control of cyclin-dependent kinase (CDK). These results indicate that the spatiotemporal regulation of microtubule formation is established by the Alp7/TACC–Alp14/TOG complex through the coordinated interplay of Ran and CDK

Macrophage centripetal migration drives spontaneous healing process after spinal cord injury.

Traumatic spinal cord injury (SCI) brings numerous inflammatory cells, including macrophages, from the circulating blood to lesions, but pathophysiological impact resulting from spatiotemporal dynamics of macrophages is unknown. Here, we show that macrophages centripetally migrate toward the lesion epicenter after infiltrating into the wide range of spinal cord, depending on the gradient of chemoattractant C5a. However, macrophages lacking interferon regulatory factor 8 (IRF8) cannot migrate toward the epicenter and remain widely scattered in the injured cord with profound axonal loss and little remyelination, resulting in a poor functional outcome after SCI. Time-lapse imaging and P2X/YRs blockade revealed that macrophage migration via IRF8 was caused by purinergic receptors involved in the C5a-directed migration. Conversely, pharmacological promotion of IRF8 activation facilitated macrophage centripetal movement, thereby improving the SCI recovery. Our findings reveal the importance of macrophage centripetal migration via IRF8, providing a novel therapeutic target for central nervous system injury

A systematic review of nonrandomized controlled trials on the curative effects of aquatic exercise

Hiroharu Kamioka1, Kiichiro Tsutani2, Yoshiteru Mutoh3, Hiroyasu Okuizum4, Miho Ohta5, Shuichi Handa4, Shinpei Okada6, Jun Kitayuguchi7, Masamitsu Kamada7, Nobuyoshi Shiozawa8, Sang-Jun Park4, Takuya Honda4, Shoko Moriyama41Faculty of Regional Environment Science, Tokyo University of Agriculture, Tokyo, Japan; 2Department of Drug Policy and Management, Graduate School of Pharmaceutical Sciences, 3Department of Physical and Health Education, Graduate School of Education, The University of Tokyo, Tokyo, Japan; 4Mimaki Onsen (Spa) Clinic, Tomi City, Japan; 5Laboratory of Aqua, Health, and Sports Medicine, 6Physical Education and Medicine Research Foundation, Nagano, Japan; 7Physical Education and Medicine Research Center Unnan, Unnan City, Japan; 8Department of Longevity and Social Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, JapanBackground: The objectives of this review were to integrate the evidence of curative effects through aquatic exercise and assess the quality of studies based on a review of nonrandomized controlled trials (nRCTs).Methods: Study design was a systematic review of nonrandomized controlled trials. Trials were eligible if they were nonrandomized clinical trials. Studies included one treatment group in which aquatic exercise was applied. We searched the following databases from 2000 up to July 20, 2009: MEDLINE via PubMed, CINAHL, and Ichushi-Web.Results: Twenty-one trials met all inclusion criteria. Languages included were English (N = 9), Japanese (N = 11), and Korean (N = 1). Target diseases were knee and/or hip osteoarthritis, poliomyelitis, chronic kidney disease, discomforts of pregnancy, cardiovascular diseases, and rotator cuff tears. Many studies on nonspecific disease (healthy participants) were included. All studies reported significant effectiveness in at least one or more outcomes. However results of evaluations with the TREND and CLEAR-NPT checklists generally showed a remarkable lack of description in the studies. Furthermore, there was the problem of heterogeneity, and we were therefore not able to perform a meta-analysis.Conclusion: Because there was insufficient evidence on aquatic exercise due to poor methodological and reporting quality and heterogeneity of nRCTs, we were unable to offer any conclusions about the effects of this intervention. However, we were able to identify problems with current nRCTs of aquatic exercise, and propose a strategy of strengthening study quality, stressing the importance of study feasibility as a future research agenda objective.Keywords: aquatic exercise, systematic review, nonrandomized controlled trials&nbsp

Transient Receptor Potential 1 Regulates Capacitative Ca2+ Entry and Ca2+ Release from Endoplasmic Reticulum in B Lymphocytes〉

Capacitative Ca2+ entry (CCE) activated by release/depletion of Ca2+ from internal stores represents a major Ca2+ influx mechanism in lymphocytes and other nonexcitable cells. Despite the importance of CCE in antigen-mediated lymphocyte activation, molecular components constituting this mechanism remain elusive. Here we demonstrate that genetic disruption of transient receptor potential (TRP)1 significantly attenuates both Ca2+ release-activated Ca2+ currents and inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ release from endoplasmic reticulum (ER) in DT40 B cells. As a consequence, B cell antigen receptor–mediated Ca2+ oscillations and NF-AT activation are reduced in TRP1-deficient cells. Thus, our results suggest that CCE channels, whose formation involves TRP1 as an important component, modulate IP3 receptor function, thereby enhancing functional coupling between the ER and plasma membrane in transduction of intracellular Ca2+ signaling in B lymphocytes

Macrophage numbers in the marginal area of sarcomas predict clinical prognosis

Even when treated comprehensively by surgery, chemotherapy, and radiotherapy, soft-tissue sarcoma has an unfavorable outcome. Because soft-tissue sarcoma is rare, it is the subject of fewer clinicopathological studies, which are important for clarifying pathophysiology. Here, we examined tumor-associated macrophages in the intratumoral and marginal areas of sarcomas to increase our knowledge about the pathophysiology. Seventy-five sarcoma specimens (not limited to a single histological type), resected at our institution, were collected, and the number of CD68-, CD163-, and CD204-positive macrophages in the intratumoral and marginal areas was counted. We then performed statistical analysis to examine links between macrophage numbers, clinical factors, and outcomes. A high number of macrophages positive for all markers in both areas was associated with worse disease-free survival (DFS). Next, we divided cases according to the FNCLCC classification (Grade 1 and Grades 2/3). In the Grade 1 group, there was no significant association between macrophage number and DFS. However, in the Grade 2/3 group, high numbers of CD163- and CD204-positive macrophages in the marginal area were associated with poor DFS. By contrast, there was no significant difference between the groups with respect to high or low numbers of CD68-, CD163-, or CD204-positive macrophages in the intratumoral area. Multivariate analysis identified the number of CD163- and CD204-positive macrophages in the marginal area as an independent prognostic factor. Macrophage numbers in the marginal area of soft-tissue sarcoma may better reflect clinical behavior

A cell-based high-throughput screening method to directly examine transthyretin amyloid fibril formation at neutral pH

Transthyretin (TTR) is a major amyloidogenic protein associated with hereditary (ATTRm) and nonhereditary (ATTRwt) intractable systemic transthyretin amyloidosis. The pathological mechanisms of ATTR-associated amyloid fibril formation are incompletely understood, and there is a need for identifying compounds that target ATTR. C-terminal TTR fragments are often present in amyloid-laden tissues of most patients with ATTR amyloidosis, and on the basis of in vitro studies, these fragments have been proposed to play important roles in amyloid formation. Here, we found that experimentally-formed aggregates of full-length TTR are cleaved into C-terminal fragments, which were also identified in patients' amyloid-laden tissues and in SH-SY5Y neuronal and U87MG glial cells. We observed that a 5-kDa C-terminal fragment of TTR, TTR81–127, is highly amyloidogenic in vitro, even at neutral pH. This fragment formed amyloid deposits and induced apoptosis and inflammatory gene expression also in cultured cells. Using the highly amyloidogenic TTR81–127 fragment, we developed a cell-based high-throughput screening method to discover compounds that disrupt TTR amyloid fibrils. Screening a library of 1280 off-patent drugs, we identified two candidate repositioning drugs, pyrvinium pamoate and apomorphine hydrochloride. Both drugs disrupted patient-derived TTR amyloid fibrils ex vivo, and pyrvinium pamoate also stabilized the tetrameric structure of TTR ex vivo in patient plasma. We conclude that our TTR81–127–based screening method is very useful for discovering therapeutic drugs that directly disrupt amyloid fibrils. We propose that repositioning pyrvinium pamoate and apomorphine hydrochloride as TTR amyloid-disrupting agents may enable evaluation of their clinical utility for managing ATTR amyloidosis

チイキザイジュウコウレイシャ ニ オケル テントウヨボウジココウリョクカン ト HDL コレステロール オヨビ シンタイノウリョク トノ カンレン コスゲムラオウダンケンキュウ

背景 : 本研究の目的は,転倒予防自己効力感とHDLコレステロール及び身体能力との関連性を調べることであった。方法 : 対象者は山梨県小菅村に在住する65歳以上の高齢者339名で,本調査には132名(38.9%)が参加した。移動能力として10m全力歩行,最大一歩幅,40cm踏台昇降を,バランス能力としてつぎ足歩行を測定した。血液性状の中で,主要なアウトカムとして,HDLコレステロールを用いた。転倒予防自己効力感は,10項目,4リッカートスケールからなる質問紙で評価した。研究デザインは,転倒恐怖の有無による2群間比較の横断研究である。結果 : 転倒予防自己効力感とHDLコレステロールの間に有意な関連はなかったが,転倒予防自己効力感が低い群では有意に移動能力が低かった。また,膝痛や腰痛を伴う男性では,転倒予防自己効力感が低かった。結論 : 自己効力感とHDLコレステロールとの関連は認められなかったが,本研究は地域在住で独立生活を営む高齢者に限定されているため,今後,良くデザインされた大規模な観察研究が必要だと考えられた。Background : The aim of the current study was to clarify relationships between self-efficacy and high-density lipoprotein cholesterol (HDL-C) and physical strength.Methods : Target participants were 339 elderly residents aged 65 years or older from Kosuge Village, Yamanashi Prefecture in Japan. One hundred and thirty two persons participated in the study (38.9%). Outcome measurements included the 10-m walking time, the maximal step length and 40-cm step test as moving ability, the tandem gait as balance ability, and HDL-C. Fall-prevention self-efficacy (FPSE) was evaluated using a questionnaire that examined 10 items (actions) and 4 Likert scale. A total of 40 points could be awarded. This research design was a cross-sectional study that divided the elderly into two groups by the existence of a fear of falling.Results : Although there was no significant relationship between fall-prevention self-efficacy and HDL-C, we demonstrated that a group with poor self-efficacy had significantly deteriorated moving ability and that only one male participant with knee pain and/or lumbago exhibited poor self-efficacy.Conclusions : We could not find significant relationships between self-efficacy and HDL-C. But this study is limited to the independent elderly people in a local area, and further studies should be conducted to detect the relationships

ジョセイカイゴシャ ニ オケル ヨウツウ ノ ジッタイ ト カンレンヨウイン ニ カンスル オウダンケンキュウ

本研究は,横断研究により,女性介護者における腰痛との関連要因を明らかにすることを目的とした。4カ所の特別養護老人ホームに勤務する女性介護職員88人を対象として,腰痛の有無,経験年数,Body Mass Index(BMI),握力,長座体前屈,指床間距離(Finger-Floor Distance : FFD),抑うつ自己評価スコア(the Center for Epidemiologic Study Depression Scale : CES-D),SF-8による身体的健康度(Physical Health Score : PHS)と精神的健康度(Mental Health Score : MHS),疲労の自覚症状を調査した。腰痛の有無を目的変数とし,その他の変数を説明変数とした多重ロジスティック回帰分析を行った。49人(55.7%)が,慢性的な腰痛を訴えていた。多重ロジスティック回帰分析の結果,腰痛の発生は,身長がオッズ比(OR)1.12,95%信頼区間(95%CI)1.02-1.23,SF-8による身体的健康度(PHS)が,OR : 0.89(95%CI : 0.86-0.97),精神的健康度(MHS)がOR : 0.90(95%CI : 0.81-0.99)で有意であった。本研究は,身長が高いことは中腰姿勢を助長し,腰痛を起こす原因になるかもしれないこと,身体・精神的な健康の程度は,腰痛と関係があることが明らかになった。今後,交代勤務に伴う睡眠障害や慢性疲労との関連性も含めて検討すべきであることが示唆された。The objective of this cross-sectional study was to reveal the actual conditions and related factors of low back pain among female caregivers in special nursing homes for the elderly.The participants were 88 female caregivers in four facilities for the elderly. We surveyed the existence of low back pain, employment period, Body Mass Index (BMI), grip power, sitting forward flexion, Finger-Floor Distance (FFD), the Center for Epidemiologic Study Depression Scale (CES-D), Physical Health Score (PHS) and Mental Health Score (MHS) by using SF-8 and subjective symptoms of fatigue. Multinomial logistic regression model was used to investigate how far low back pain was related to mental and physical health status, flexibility, power, employment period, and BMI.55.7% (N=46) of the participants notified the presence of chronic lumbago. The result of multinomial logistic regression model showed that the existence of low back pain was significant in the following variables : 1) Odds ratio (OR)=1.12 (95% confidence interval : 95%CI=1.02-1.23) for height, 2) OR=0.89 (95% CI=0.81-0.97) for PHS, 3) OR=0.90 (95% CI=0.81-0.99) for MHS.Our findings suggest that height may reinforce a half-crouching position, thus it can be a cause of low back pain, and that the degree of physical and mental health is related to low back pain. It was also suggested that lumbago prevention and/or reduction strategy should include the relevance of sleep interruption and chronic fatigue with shift work when this problem is examined in future

Spatiotemporal Regulation of Nuclear Transport Machinery and Microtubule Organization

Spindle microtubules capture and segregate chromosomes and, therefore, their assembly is an essential event in mitosis. To carry out their mission, many key players for microtubule formation need to be strictly orchestrated. Particularly, proteins that assemble the spindle need to be translocated at appropriate sites during mitosis. A small GTPase (hydrolase enzyme of guanosine triphosphate), Ran, controls this translocation. Ran plays many roles in many cellular events: nucleocytoplasmic shuttling through the nuclear envelope, assembly of the mitotic spindle, and reorganization of the nuclear envelope at the mitotic exit. Although these events are seemingly distinct, recent studies demonstrate that the mechanisms underlying these phenomena are substantially the same as explained by molecular interplay of the master regulator Ran, the transport factor importin, and its cargo proteins. Our review focuses on how the transport machinery regulates mitotic progression of cells. We summarize translocation mechanisms governed by Ran and its regulatory proteins, and particularly focus on Ran-GTP targets in fission yeast that promote spindle formation. We also discuss the coordination of the spatial and temporal regulation of proteins from the viewpoint of transport machinery. We propose that the transport machinery is an essential key that couples the spatial and temporal events in cells