研究者詳細 - 廣畑　聡

論文 - 廣畑　聡

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新規ラットlink protein geneの同定と特性 Lp3/Hapln3(Identification and Characterization of a novel rat link protein gene: Lp3/Hapln3)

小川弘子, 廣畑聡, 三好亨, 村上充, 白鳥康史, 大橋俊孝, 二宮善文, 草地省蔵, 佐田政隆

Circulation Journal 68 ( Suppl.III ) 938 - 938 2004年10月

Combination of caspase transfer using the human telomerase reverse transcriptase promoter and conventional therapies for malignant glioma cells 国際誌

H Takeuchi, T Kanzawa, Y Kondo, T Komata, S Hirohata, S Kyo, S Kondo

INTERNATIONAL JOURNAL OF ONCOLOGY 25 ( 1 ) 57 - 63 2004年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PROFESSOR D A SPANDIDOS

Recently, we have reported the therapeutic efficacy of delivering initiator caspase (caspase-8) or executioner active caspase (rev-caspase-6) to telomerase-positive malignant glioma cells using the human telomerase reverse transcriptase (hTERT) gene promoter system (hTERT/caspase-8 or hTERT/rev-caspase-6). In the present study, we investigated if conventional treatments for malignant gliomas augment the efficacy of the hTERT/caspase therapy. First, we demonstrated that hTERT/rev-caspase-6 exhibited a greater ability to induce apoptosis in malignant glioma U87-MG and U373-MG cells than hTERT/caspase-8. Next, as conventional treatments to combine with hTERT/rev-caspase-6, apoptosis-inducing agents [cisplatin (CDDP), paclitaxel (PTX), and BCNU] and non-apoptosis-inducing therapies [temozolomide (TMZ) and gamma-irradiation (IR)] were used. Combination of hTERT/rev-caspase-6 gene therapy with PTX yielded a dose-dependent additive effect, while CDDP and BCNU had additive effect only when tumor cells were treated at IC75 of each agent. A decline in the combination effect of CDDP and BCNU at IC50 was due to decreased activity of telomerase in treated tumor cells prior to the hTERT/rev-caspase-6 transfer. On the other hand, TMZ or IR had no significant additive effect on induction of apoptosis. These results suggest that agents, which induce apoptosis without inhibiting telomerase activity are a promising counterpart to combine with hTERT/rev-caspase-6 therapy for the management of malignant gliomas.

DOI： 10.3892/ijo.25.1.57

新規マトリックスリンクプロテインLp3/Hapln3 クローニングおよびその発現解析

小川弘子, 廣畑聡, 大橋俊孝, 佐田政隆, 村上充, 二宮善文, 草地省蔵, 白鳥康史, 大江透

日本動脈硬化学会総会プログラム・抄録集 36回 197 - 197 2004年7月

Spatially and temporally different expression of osteonectin and osteopontin in the infarct zone of experimentally induced myocardial infarction in rats

Komatsubara, I, T Murakami, S Kusachi, K Nakamura, S Hirohata, J Hayashi, S Takemoto, C Suezawa, Y Ninomiya, Y Shiratori

CARDIOVASCULAR PATHOLOGY 12 ( 4 ) 186 - 194 2003年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER SCIENCE INC

Osteonectin and osteopontin, two secreted matricellular proteins, have a variety of functions that are exerted through interaction with matrix components. These proteins appear in response to tissue injury. To test our hypothesis that osteopontin and osteonectin are expressed with spatially and temporally different patterns in myocardial infarct tissue, we investigated osteonectin and osteopontin expression in experimentally induced myocardial infarction in rats, in comparison with Type I collagen expression. Northern blotting demonstrated that osteonectin mRNA did not markedly increase on Day 2 after the infarction, but it increased on Days 7 and 14 by 1.7 +/- 0.12- and 1.8 +/- 0.01-fold compared to that in preligation hearts. In contrast, osteopontin mRNA was increased on Day 1 (41.9 +/- 11.3-fold increase) and on Day 2 (58.3 +/- 7.6-fold increase), and then it declined on Days 7 and 14 (24.8 +/- 9.0- and 13.5 +/- 4.7-fold increase, respectively). In situ hybridization revealed that osteonectin mRNA signals were observed in fibroblasts, myofibroblasts and macrophages around infarct necrotic tissue on Days 7 and 14. Osteopontin mRNA signals were observed in macrophages in the infarct marginal zone on Day 2. Immunopositive staining for both osteonectin and osteopontin showed the same pattern as that obtained by in situ hybridization. The time course of osteonectin mRNA was almost parallel with that of Type I collagen mRNA, while that of osteopontin was not. These results demonstrated spatially and temporally different expression patterns of osteonectin and osteopontin in myocardial infarction and suggest that osteonectin appears to be involved in the pathological course in the late phase after infarction concomitantly with Type I collagen, while osteopontin may play a role in the early phase. (C) 2003 Elsevier Inc. All rights reserved.

DOI： 10.1016/S1054-8807(03)00042-5

Osteogenic protein-1 reduces intercellular adhesion molecule-1 messenger RNA expression, infarct size and TUNEL-positive cardiomyocytes in ischemia/reperfusion rat hearts. 査読国際誌

Hayashi J, Kusachi S, Murakami T, Miyoshi T, Nakamura K, Koten K, Ogawa H, Hirohata S, Ninomiya Y, Shiratori Y

Experimental and clinical cardiology 8 ( 4 ) 195 - 200 2003年

胆嚢炎に続発した感染性腹部大動脈瘤の一例

三好亨, 広畑敦, 小天和也, 土井正行, 廣畑聡, 瀬崎悟之, 村上充, 草地省蔵, 白鳥康史

Circulation Journal 2003年

Intense response of heart rate with pronounced suppression of high-frequency power of heart rate variability to early morning exercise with high-intensity load 査読国際誌

J Ueta, T Nakatsu, T Murakami, S Toyonaga, S Hirohata, K Mashima, M Sangawa, S Kusachi, Y Shiratori

BIOMEDICINE & PHARMACOTHERAPY 56 ( SUPPL. 2 ) 353S - 358S 2002年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

Autonomic nerve activity shows circadian variation. Therefore, we put forward the hypothesis that the responses of heart rate (HR) and high-frequency (HF) power of HR variability to exercise would be different between-early morning and daytime exercise. We performed ergometer constant load exercise tests [50 watts (low), 100 watts (high load)] in the early morning and during the day in 6 healthy volunteers. The HR response was fitted to an exponential hyperbolic sine function: HR = alpha*e(-beta*t) *sinh(omega*t)+gamma. In this equation, the beta/omega ratio was inversely correlated with the intensity of the HR response. HF power was determined using a recently developed algorithm with high time-resolution power. There were no significant differences in HR, HF power or systolic blood pressure (BP) pressure before exercise between early morning and daytime exercise with either the 50 or 100 watt loads. During exercise, there were no significant differences in maximal HR or maximal systolic BP between early morning and daytime exercise with either 50 or 100 watt loads. For high-load exercise, the beta/omega ratio was significantly lower in early morning exercise (mean+/-SD, 0.945+/-0.02) than in daytime exercise (0.987+/-0.03). Similarly, for 100 watt exercise, HF power of HR variability was significantly lower in early morning exercise (0.94+/-0.52 msec[Hz(1/2)) than in daytime exercise (1.26+/-0.74 msec/Hz(1/2)). In conclusion, the present study demonstrated that a lower beta/omega ratio in the HR response was associated with lower HF power of HR variability in early morning high-load exercise compared to that in daytime exercise, indicating that the heart rate responded more intensely to early morning exercise than to daytime exercise with a high load due, at least partly, to pronounced suppression of parasympathetic nerve activity. (C) 2002 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

DOI： 10.1016/s0753-3322(02)00316-5

A novel treatment of human malignant gliomas in vitro and in vivo: FADD gene transfer under the control of the human telomerase reverse transcriptase gene promoter

T Komata, S Koga, S Hirohata, M Takakura, IM Germano, M Inoue, S Kyo, S Kondo, Y Kondo

INTERNATIONAL JOURNAL OF ONCOLOGY 19 ( 5 ) 1015 - 1020 2001年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PROFESSOR D A SPANDIDOS

Telomerase activity has a close relationship with malignancies in many cell types and it is tightly regulated at the transcriptional level of human telomerase reverse transcriptase (hTERT). Utilizing the hTERT promoter, the authors developed a gene delivery system of Fas associated protein with death domain (FADD) (hTERT/FADD). FADD is a protein which plays an important role in the apoptotic pathway of Fas. Overexpression of FADD induces apoptosis in the cells regardless of Fas expression on the cell surface. We hypothesized that we might be able to restrict the expression of FADD in malignant glioma cells if we use the gene transfer system under the control of hTERT promoter. This study was designed to investigate whether the hTERT/FADD construct induces apoptosis effectively in malignant glioma cells while keeping normal cells intact. First, using the reverse transcription-polymerase chain reaction (RT-PCR) technique, we confirmed that hTERT mRNA was expressed in human malignant glioma cells (U373-MG, A172 and GB-1), but not in cultured astrocytes (TEN) or fibroblasts (MRC5). After transient transfection with the hTERT/FADD construct, a significant number of FADD-positive cells and apoptotic cells were detected in hTERT-positive malignant glioma cells. In contrast, hTERT-negative astrocytes and fibroblasts remained intact. Furthermore, subcutaneously implanted U373-MG tumors treated with the hTERT/FADD construct reduced in volume significantly C compared to the conrol treatment (p=0.0001). Gene transfer of FADD under the control of the hTERT promoter may be a novel and promising therapy to kill hTERT-positive malignant glioma cells while sparing normal brain cells lacking hTERT.

DOI： 10.3892/ijo.19.5.1015

Increased expression of biglycan mRNA in pressure-overloaded rat heart

Y Ayada, S Kusachi, T Murakami, S Hirohata, S Takemoto, Komatsubara, I, J Hayashi, A Iwabu, Y Ninomiya, T Tsuji

CLINICAL AND EXPERIMENTAL HYPERTENSION 23 ( 8 ) 633 - 643 2001年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MARCEL DEKKER INC

Biglycan mRNA expression in rat myocardium after abdominal aortic banding with renal ischemia was examined. The Northern blot analysis demonstrated that expression of biglycan mRNA in the pressure-overloaded hearts on days 2, 7, 14 and 28 was 2.88 +/-0.89, 2.32 +/-0.49, 2.17 +/-0.57 and 1.81 +/-0.46-fold higher, respectively, than that in the sham-operated hearts. In situ hybridization showed an increased density of biglycan mRNA signal-positive cells in the pressure-overloaded hearts. The cells with positive signals were spindle-shaped mesenchymal cells in the myocardial interstitium. A marked increase in biglycan mRNA signal expression was also observed in endothelial cells and smooth muscle cells of the thickened myocardial capillary wall. These results demonstrated an increase in biglycan mRNA in the pressure-overloaded heart in mesenchymal cells in the myocardial interstitium, and in endothelial and smooth muscle cells of the capillaries, indicating that biglycan contributes to the ventricular and vascular remodeling in response to pressure overload.

DOI： 10.1081/CEH-100107393

ラット心筋梗塞モデルでの新生血管におけるオステオネクチンmRNAの発現の経時的変化の検討

前田弘子, 村上充, 廣畑聡, 中村圭吾, 岩部明弘, 綾田陽子, 瀬崎悟之, 竹本俊二, 小松原一正, 草地省蔵

Japanese Circulation Journal 65 ( Suppl.III ) 807 - 807 2001年10月

ラット実験的心筋梗塞モデルにおけるトロンボスポンジン-1mRNA発現の経時的変化の検討

中村圭吾, 廣畑聡, 岩部明弘, 綾田陽子, 前田弘子, 竹本俊二, 瀬崎悟之, 小松原一正, 十枝健一, 村上充

マトリックス研究会大会 ( 48 ) 70 - 70 2001年4月

Spatial changes of gelatinolytic activity in myocardial infarction in rats

Takashi Murakami, Shozo Kusachi, Satoshi Hirohata, Chisato Suezawa, Syunji Takemoto, Satoshi Sezaki, Takao Tsuji, Yoshifumi Ninomiyal

Keio Journal of Medicine 50 54 - 63 2001年1月

新しい細胞外マトリックス分解酵素ＡＤＡＭＴＳファミリー

廣畑聡

生化学 73,11,1333-1337 2001年

Chromosomal assignment of two ADAM genes, TACE (ADAM17) and MLTNB (ADAM19), to human chromosomes 2 and 5, respectively, and of Mltnb to mouse chromosome 11.

Genomics. Nov 15;54(1):178-9 ( 1 ) 178 - 179 1998年11月

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