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Ca2+-Induced Conformational Change of Troponin C from the Japanese Pearl Oyster, Pinctada fucata

American Journal of Molecular Biology 2018年第4期8卷 205-214页

作者： daisuke Funabara daisuke ishikawa Yoshinori Urakawa Satoshi Kanoh Graduate School of Bioresources Mie University Tsu Japan

Troponin is a thin filament-associated regulator of vertebrate striated muscle contraction. Troponin changes its structure upon Ca2+ binding to troponin C, one of the subunits of troponin, allowing myosin to interact with actin. We recently elucidated the molecular characteristics of the Japanese pearl oyster Pinctada fucata troponin C (Pifuc-TnC), revealing the possibilities that Pifuc-TnC and vertebrate muscle TnC play dissimilar roles in muscle contraction. Pifuc-TnC has four EF-hand motifs, but, unlike vertebrate TnC, only one (site IV) was predicted to bind Ca2+. To confirm the number of Ca2+-binding sites in Pifuc-TnC and whether Ca2+ binding induces a conformational change, we purified the full-length protein and a variant, Pifuc-TnC-E142Q (that has a mutation in the predicted Ca2+-binding site of site IV), following their expression in laboratory E. coli. Isothermal titration calorimetry demonstrated Ca2+ binding to Pifuc-TnC, whereas Pifuc-TnC-E142Q was unable to bind Ca2+, confirming that site IV is the only Ca2+-binding site in Pifuc-TnC. Pifuc-TnC eluted in a later fraction from a gel filtration column in the presence of Ca2+ compared with the condition when Ca2+ was absent. In contrast, the elution profiles of Pifuc-TnC-E142Q were equivalent in both the presence and absence of Ca2+, suggesting that Ca2+ binding to Pifuc-TnC induces a conformational change that delays its elution from the column. UV-absorption spectral analysis revealed that binding of Ca2+ to Pifuc-TnC caused an increase in absorption at a wavelength of approximately 250 nm, possibly because phenylalanine residues had been exposed on the surface of the molecule as a result of a conformational change. Differential scanning calorimetric analyses of Pifuc-TnC showed aggregation in the presence of Ca2+ in accordance with an increase of temperature, but no aggregation was seen in the absence of Ca2+. In combination, these findings suggest that Ca2+ binding to site IV induces a conformational

关键词： Ca2+-Binding Catch Muscle Conformational Change EF-Hand Troponin C

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Troponin T from the Japanese Pearl Oyster Pinctada fucata: Molecular Cloning, Tissue Distribution, Gene Structure, and Interaction Analysis with Tropomyosin

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American Journal of Molecular Biology 2020年第1期10卷 61-73页

作者： daisuke Funabara Yoshinori Urakawa daisuke ishikawa Satoshi Kanoh Graduate School of Bioresources Mie University Tsu Japan

Troponin (Tn) is composed of three subunits (TnI, TnC and TnT) that bind Ca2+ and regulate striated muscle contraction in vertebrates. TnT’s function has been extensively described in vertebrates, but its role has been obscure in molluscan muscles. Our previous work indicated that the TnC and TnI subunits work in adductor phasic muscle, but not in catch muscle. Here, we have characterized TnT from the Japanese bivalve pearl oyster Pinctada fucata to start to explain the function of Tn in molluscan muscle contraction. We determined the primary structure of the full-length TnT protein from the P. fucata adductor muscle (Pifuc-TnT), and found that it is composed of 316 amino acid residues with a predicted molecular mass of 37.4 kDa. Multiple sequence alignment showed that Pifuc-TnT has an extension of >60 residues at the C-terminus that are not present in vertebrate TnTs, including known TnTs from other mollusks. Pifuc-TnT gene structure predictions using Splign alignment of the cDNA generated in this study and genome sequences indicated that Pifuc-TnT consists of 13 exons. Start and stop codons are located in exons 2 and 12, respectively. Quantitative real-time PCR revealed that the Pifuc-TnT gene was predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, slightly in gill, and not at all in mantle and foot. These findings suggest that TnT plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction. Isothermal titration calorimetry revealed that unlike vertebrate TnTs, Pifuc-TnT does not interact with P. fucata tropomyosin-1 nor with tropomyosin-2. These findings in P. fucata imply that Tn functions differently in molluscan muscle than it does in vertebrates.

关键词： Adductor Muscle Catch Contraction Pinctada fucata Troponin T Tropomyosin

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Magnifying chromoscopy, a novel and useful technique for colonoscopy in ulcerative colitis

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World Journal of Gastroenterology 2007年第18期13卷 2523-2528页

作者： Takafumi Ando Hironao Takahashi Osamu Watanabe Osamu Maeda Kazuhiro Ishiguro daisuke ishikawa Motofusa Hasegawa Naoki Ohmiya Yasumasa Niwa Hidemi Goto Naoki Ohmiya Yasumasa Niwa Hidemi Goto Department of Gastroenterology Nagoya University Graduate School of Medicine Nagoya Japan

Ulcerative colitis (UC) is a chronic inflammatory bowel disorder characterized by exacerbations and remissions. The degree of inflammation as assessed by conventional colonoscopy is a reliable parameter of disease activity. However, even when conventional colonoscopy suggests remission and normal mucosal findings, microscopic abnormalities may persist, and relapse may occur later. Patients with long-standing, extensive ulcerative colitis have an increased risk of developing colorectal cancer. Ulcerative colitis-associated colorectal cancer is characterized by an early age at onset, poorly differentiated tumor cells, mucinous carcinoma, and multiple lesions. Early detection of dysplasia and colitic cancer is thus a prerequisite for survival. A relatively new method, magnifying chromoscopy, is thought to be useful for the early detection and diagnosis of dysplasia and colitic cancer, as well as the prediction of relapse.

关键词： Ulcerative colitis Histopathology Conventionalcolonoscopy Magnifying colonoscopy Ulcerative colitis-associated colorectal cancer

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Value of colonoscopy for prediction of prognosis in patients with ulcerative colitis

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World Journal of Gastroenterology 2008年第14期14卷 2133-2138页

作者： Takafumi Ando Yuji Nishio Osamu Watanabe Hironao Takahashi Osamu Maeda Kazuhiro Ishiguro daisuke ishikawa Naoki Ohmiya Yasumasa Niwa Hidemi Goto Department of Gastroenterology Nagoya University Graduate School of Medicine Nagoya 4668550 Japan Department of Gastroenterology Nagoya University Graduate School of Medicine Nagoya 4668550 Japan Department of Gastroenterology Meitetsu Hospital Nagoya 4518511 Japan

Ulcerative colitis (UC) is a chronic inflammatory bowel disorder characterized by exacerbations and remissions. Some UC patients remain refractory to conventional medical treatment while, in others, the effectiveness of drugs is limited by side-effects. Recently, cyclosporine and leukocyte removal therapy have been used for refractory UC patients. To predict the efficacy of these therapies is important for appropriate selection of treatment options and for preparation for colectomy. Endoscopy is the cornerstone for diagnosis and evaluation of UC. Endoscopic parameters in patients with severe or refractory UC may predict a clinical response to therapies, such as cyclosporine or leukocyte removal therapy. As for the patients with quiescent UC, relapse of UC is difficult to predict by routine colonoscopy. Even when routine colonoscopy suggests remission and a normal mucosal appearance, microscopic abnormalities may persist and relapse may occur later. To more accurately identify disease activity and to predict exacerbations in UC patients with clinically inactive disease is important for deciding whether medical treatment should be maintained. Magnifying colonoscopy is useful for the evaluation of disease activity and for predicting relapse in patients with UC.

关键词： Ulcerative colitis Colonoscopy Prediction of outcome

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Hyperacute experimental model of rat lung transplantation using a coronary shunt cannula

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World Journal of Transplantation 2024年第2期14卷 155-161页

作者： Munehisa Takata Yusuke Tanaka daisuke Saito Shuhei Yoshida Isao Matsumoto Department of Thoracic Surgery Kanazawa UniversityKanazawa 920-8641IshikawaJapan

BACKGROUND Lung transplantation is a well-established treatment of end-stage lung disease.A rodent model is an inexpensive way to collect biological data from a living model after lung ***,mastering the surgical technique takes time owing to the small organ *** To conduct rat lung transplantation using a shunt cannula(SC)or modified cannula(MC)and assess their *** Rat lung transplantation was performed in 11 animals in the SC group and 12 in the MC *** devised a method of rat lung transplantation using a coronary SC for coronary artery bypass surgery as an anastomosis of pulmonary arteri-ovenous vessels and *** same surgeon performed all surgical proce-dures in the donor and recipient rats without using a magnifying *** success rate of lung transplantation,operating time,and PaO2 values were com-pared after 2-h reperfusion after *** Ten and 12 lungs were successfully transplanted in the SC and MC groups,*** the SC group,one animal had cardiac arrest within 1 h after reperfusion owing to bleeding during pulmonary vein *** opera-ting time for the removal of the heart-lung block from the donor and preparation of the left lung graft was 26.8±2.3 and 25.7±1.3 min in the SC and MC groups,respectively(P=0.21).The time required for left lung transplantation in the recipients was 37.5±2.8 min and 35.9±1.4 min in the SC and MC groups,respectively(P=0.12).PaO2 values at 2 h after reperfusion were 456.2±25.5 and INTRODUCTION Lung transplantation is a well-established treatment of end-stage lung *** immune and non-immune mech-anisms in lung transplantation are highly complex,and post-transplant complications such as infections and primary and chronic lung allograft dysfunction must be reduced to improve ***,there is a need for immunological and pathophysiological analyses using animal lung transplantation *** rat lung transplantation model was f

关键词： Lung transplantation Rat Shunt cannula Modified cannula Reperfusion

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Can robotic liver resection compensate for weaknesses of the laparoscopic approach?

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Hepatobiliary Surgery and Nutrition 2020年第3期9卷 385-387页

作者： daisuke Ban Yoshiya ishikawa Minoru Tanabe Department of Hepatobiliary and Pancreatic Surgery Graduate School of MedicineTokyo Medical and Dental UniversityTokyoJapan

In the past decade,the induction of minimal invasive surgery(MIS)has revolutionized liver surgery,because this approach greatly improves the clinical course of postoperative patients(1).

关键词： surgery liver clinical

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Formation mechanism of extension fractures induced by excavation of a gallery in soft sedimentary rock,Horonobe area,Northern Japan

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Geoscience Frontiers 2013年第1期4卷 105-111页

作者： Tetsuya Tokiwa Kimikazu Tsusaka Makoto Matsubara Taiki ishikawa daisuke Ogawa Horonobe Underground Research Unit Japan Atomic Energy AgencyHokkaido 098-3224Japan Geoscience Research Laboratory Kanagawa 242-0014Japan Mitsubishi Materials Techno Corporation Tokyo 102-8205Japan

This paper focuses on the formation mechanism of fractures induced by excavation of a gallery in soft sedimentary rocks in the Horonobe area of Japan. Detailed fracture mapping of the gallery indicates that the fractures consist of both pre-existing shear fractures and excavation damaged zone （EDZ） fractures. EDZ fractures correspond to weak planes associated with bedding planes or transgranular cracks. The EDZ fractures terminate against pre-existing shear fractures. Therefore, even for excavations in soft sedimentary rocks, formation of the EDZ fractures are controlled by pre-existing fractures and earlier weak planes.

关键词： Fracture Excavation damaged zone (EDZ)Weak plane Soft rock Excavation

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Overexpression of c-met in the early stage of pancreatic carcinogenesis; altered expression is not sufficient for progression from chronic pancreatitis to pancreatic cancer

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World Journal of Gastroenterology 2006年第24期12卷 3878-3882页

作者： Kenoki Ohuchida Kazuhiro Mizumoto Nami ishikawa Yasuhiro Ogura daisuke Yamada Takuya Egami Hayato Fujita Seiji Ohashi Eishi Nagai Masao Tanaka Department of Surgery and Oncology Graduate School of Medical Sciences Kyushu UniversityFukuokaJapan

AIM： To investigate c-met expression during early pancreatic carcinogenesis. METHODS： We used 46 bulk tissues and 36 microdissected samples, including normal pancreas, chronic pancreatitis, and pancreatic cancer, for quantitative realtime reverse transcription-polymerase chain reaction. RESULTS： In bulk tissue analyses, pancreatic cancer tissues expressed significantly higher levels of c-met than did chronic pancreatitis and normal pancreas tissues. c-met levels did not differ between chronic pancreatitis and normal pancreas tissues. In microdissection-based analyses, c-met was expressed at higher levels in microclissected pancreatic cancer cells and pancreatitisaffected epithelial cells than in normal ductal epithelial cells （both, P 〈 0.01）. Interestingly, pancreatitis-affected epithelial cells expressed levels of c-met similar to those of pancreatic cancer cells. CONCLUSION： Overexpression of c-met occurs during the early stage of pancreatic carcinogenesis, and a single alteration of c-met expression is not sufficient for progression of chronic pancreatitis-affected epithelial cells to pancreatic cancer cells.

关键词： c-met', Pancreatic cancer Chronic pancreatitis Pancreatic carcinogenesis

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Double-balloon endoscopic retrograde cholangiopancreatography for patients who underwent liver operation: A retrospective study

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World Journal of Gastroenterology 2020年第10期26卷 1056-1066页

作者： Ryo Nishio Hiroki Kawashima Masanao Nakamura Eizaburo Ohno Takuya ishikawa Takeshi Yamamura Keiko Maeda Tsunaki Sawada Hiroyuki Tanaka daisuke Sakai Ryoji Miyahara Masatoshi Ishigami Yoshiki Hirooka Mitsuhiro Fujishiro Department of Gastroenterology and Hepatology Nagoya University Graduate School of MedicineAichi PrefectureNagoya 4668550Japan Department of Endoscopy Nagoya University HospitalAichi PrefectureNagoya 4668550Japan Department of Liver Biliary Tract and Pancreas DiseasesFujita Health University HospitalAichi PrefectureToyoake 4701192Japan

BACKGROUND Double-balloon endoscopic retrograde cholangiography (DB-ERC) is widely performed for biliary diseases after reconstruction in gastrointestinal surgery,but there are few reports on DB-ERC after hepatectomy or living donor liver transplantation (LDLT).AIM To examine the success rates and safety of DB-ERC after hepatectomy or LDLT METHODS The study was performed retrospectively in 26 patients (45 procedures) who underwent hepatectomy or LDLT (liver operation:LO group) and 40 control patients (59 procedures) who underwent pancreatoduodenectomy (control group).The technical success (endoscope reaching the choledochojejunostomy site),diagnostic success (performance of cholangiography),therapeutic success(completed interventions) and overall success rates,insertion and procedure(completion of DB-ERC) time,and adverse events were compared between these *** There were no significant differences between LO and control groups in the technical [93.3%(42/45) vs 96.6%(57/59),P=0.439],diagnostic [83.3%(35/42) vs83.6%(46/55),P=0.968],therapeutic [97.0%(32/33) vs 97.7%(43/44),P=0.836],and overall [75.6%(34/45) vs 79.7%(47/59),P=0.617] success *** median insertion time (22 vs 14 min,P <0.001) and procedure time (43.5 vs 30 min,P=0.033) were significantly longer in the LO *** incidence of adverse events showed no significant difference [11.1%(5/45) vs 6.8%(4/59),P=0.670].CONCLUSION DB-ERC after liver operation is safe and useful but longer time is required,so should be performed with particular care.

关键词： Biliary tract diseases Double-balloon enteroscopy Endoscopic retrograde cholangiopancreatography Hepatectomy Liver transplantation Risk management

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Spinal Epidural Myeloid Sarcoma with Paraplegia in Acute Myeloid Leukemia Treated with Radiation and Chemotherapy: A Case Report

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Open Journal of Orthopedics 2019年第6期9卷 131-136页

作者： Chiaki Sato Naohisa Miyakoshi Michio Hongo Yuji Kasukawa Yoshinori ishikawa daisuke Kudo Yoichi Shimada Department of Orthopedic Surgery Akita University Graduate School of Medicine Akita Japan

Myeloid sarcoma occurs in 1% - 9% of patients with myelogenous leukemia. Spinal epidural myeloid sarcoma is particularly rare, and its treatment has not been established. A 27-year-old woman complained of pain on her left chest, back around the scapula, and neck. Magnetic resonance imaging (MRI) showed a thoracic epidural tumor. One week after her visit, she developed motor weakness of her lower extremities and dysuria, and she was diagnosed with acute myelogenous leukemia (AML) on peripheral blood analysis. The epidural tumor was strongly suspected to be myeloid sarcoma. The paralysis of the lower extremities and bladder dysfunction were not progressive, and chemotherapy and local radiation therapy to the spine were performed. Improvement of paralysis and complete reduction of tumor volume were achieved by the combination of local low-dose radiation therapy and chemotherapy.

关键词： Myeloid Sarcoma Acute Myeloid Leukemia Spinal Cord Compression

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