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151. Disinfection potential of electrolyzed solutions containing sodium chloride at low concentrations

Author

Morita, Chizuko, primary, Sano, Kouichi, additional, Morimatsu, Shinichi, additional, Kiura, Hiromasa, additional, Goto, Toshiyuki, additional, Kohno, Takehiro, additional, Hong, Wu, additional, Miyoshi, Hirofumi, additional, Iwasawa, Atsuo, additional, Nakamura, Yoshiko, additional, Tagawa, Masami, additional, Yokosuka, Osamu, additional, Saisho, Hiromitsu, additional, Maeda, Toyoyuki, additional, and Katsuoka, Yoji, additional

Published
2000
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179. The observation of UEA-I binding sites of stomach by adjacent semithin and ultrathin sections.

Author

TSUYAMA, Shinichiro, primary, IHIDA, Kaori, additional, KASHIO, Nobuyuki, additional, MURATA, Fusayoshi, additional, Tsuzuki, Hideaki, additional, Imamura, Yoshiaki, additional, Noriki, Sakon, additional, Fukuda, Masaru, additional, UEDA, Takashi, additional, TERADA, Satoshi, additional, YAMADA, Kazuyori, additional, UEMURA, Yoshiko, additional, Ueno, Masaki, additional, Nakamura, Yoshiko, additional, Umezaki, K., additional, Nakajima, T., additional, Sawaragi, I., additional, Okamura, A., additional, Utsunomiya, H., additional, Sato, S., additional, Takekoshi, S., additional, Komatsu, N., additional, Suemizu, H., additional, Watanabe, K., additional, Wang, Nian, additional, WATANABE, Masahito, additional, GOTO, Hideyuki, additional, NAKATSUKA, Yasuhiko, additional, KOIDE, Tetsuya, additional, SHIMADA, Masahisa, additional, WATANABE, Sadahiro, additional, SASAKI, Junzo, additional, Yagita, Kazuhiro, additional, Okamura, Hitoshi, additional, Ibata, Yasuhiko, additional, YAMADA, Hisao, additional, KUROKAWA, Kiyoshi, additional, OCHI, Junzo, additional, YAMAZAKI, Noriko, additional, KUMAZAWA, Hirobumi, additional, Hori, Yoshiro, additional, KITA, Jun, additional, SAI, Shinichi, additional, YAMASHITA, Toshio, additional, KUMAZAWA, Tadami, additional, KAWAMOTO, Keiji, additional, YAMAZAKI, Yukinao, additional, KOHLI, Yoshihiro, additional, FUJIKI, Norio, additional, IMAMURA, Yoshiaki, additional, YANASE, Yoshizo, additional, KAKUDO, K., additional, YAMAGUCHI, T., additional, MORIKAWA, Y., additional, MATSUURA, N., additional, TAMAKI, T., additional, Yao, Chong-Hua, additional, Kitazawa, Sohei, additional, Maeda, Sakan, additional, YASUDA, Kenjiro, additional, AISO, Sadakazu, additional, SHIOZAWA, Masahide, additional, YAMASHITA, Shuji, additional, TAKEUCHI, Yuji, additional, YASUI, Hitoshi, additional, TAKAMATSU, Tetsuro, additional, FUJITA, Setsuya, additional, YOKOSE, Satoshi, additional, IKEDA, Tohru, additional, YAMAGUCHI, Akira, additional, TAJIMA, Yoshifumi, additional, UTSUMI, Nobuo, additional, YOSHIKI, Shusaku, additional, YOKOTA, Sadaki, additional, YOSHINAGA-HIRABAYASHI, Tomoko, additional, Yu, Xiang-Min, additional, Liu, You-Zhang, additional, Noda, Toru, additional, and Ogawa, Kazuo, additional

Published
1992
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181. Protective effect of estrogen combined with aspirin against re-stenosis of vein graft

Author

MATSUMURA, Akihiko, primary, NUMANO, Fujio, additional, Matsuta, Morimasa, additional, Matsuta, Mayumi, additional, Nishiya, Iwao, additional, Weier, Heinz UI., additional, Pinkel, Dan, additional, Gray, Joe W., additional, MISHIRO, Kayo, additional, WATANABE, Jun, additional, KANAMURA, Shinsuke, additional, MIYASHITA, Hiroshi, additional, SHIMADA, Kazuyuki, additional, SAITOU, Takuma, additional, MIYAYAMA, Haruhiko, additional, NISHIMURA, Reiki, additional, MIYOSHI, Norio, additional, FUKUDA, Masaru, additional, MIZUKAWA, Kiminao, additional, Mizuno, M., additional, Kagawa, K., additional, Matsumoto, M., additional, Sakabe, K., additional, Kashima, K., additional, Ashihara, T., additional, MOMIYAMA, Masayoshi, additional, FUJITA, Satoshi, additional, KONDOH, Yasumitsu, additional, KAGIYAMA, Naoto, additional, HORI, Samuel H., additional, MOMOSE, Yasunori, additional, NAGATA, Tetsuji, additional, Mori, Ichiro, additional, Watanabe, Keiichi, additional, Morii, Eiichi, additional, Nomura, Shintaro, additional, MORINO, Eri, additional, KATAOKA, Katsuko, additional, MORITA, Mika, additional, TANAKA, Kanji, additional, KAWANISHI, Hiroshi, additional, HIOKI, Koshirou, additional, MORIYAMA, Nobuo, additional, KAWABE, Kazuki, additional, HAMADA, Kozo, additional, KIMURA, Kiyoshi, additional, TAJIMA, Atsushi, additional, ASO, Yoshio, additional, MURAKOSHI, M., additional, TAGAWA, M., additional, INADA, R., additional, SHOJI, M., additional, SUZUKI, M., additional, WATANABE, K., additional, NAGASE, Yasushi, additional, TAKATA, Kuniaki, additional, HIRANO, Hiroshi, additional, NAITOH, Hiroyuki, additional, HOSOKAWA, Yohei, additional, HATTORI, Takanori, additional, NAKAJIMA, Junko, additional, NAKAZAWA, Midori, additional, KANI, Kazutaka, additional, MAEDA, Toshiko, additional, NAKAJIMA, Shigemi, additional, KUSHIMA, Ryoji, additional, Nakamura, Yoshiko, additional, Ueno, Masaki, additional, Nakazawa, Tomoko, additional, Aikawa, Eizo, additional, NISHIKAWA, Sumio, additional, HIRATA, Junko, additional, SASAKI, Fumie, additional, NISHIMURA, Rieko, additional, OKADA, Hiyoshi, additional, TSUBURA, Airo, additional, MORII, Sotokichi, additional, NISHINO, Toshihiko, additional, INAI, Shunsuke, additional, HAGA, Satomi, additional, NAGANO, Kouko, additional, YAMAMOTO, Hiroshi, additional, KIYOZUKA, Yasuhiko, additional, Nitta, Yasuo, additional, Kohli, Yoshihiro, additional, Fujiki, Norio, additional, Imamura, Yoshiaki, additional, Noriki, Sakon, additional, Fukuda, Masaru, additional, Notoya, Mitsuru, additional, Mizuhira, Vinci, additional, Hasegawa, Hiroshi, additional, ODA, K, additional, UMEMURA, S, additional, ITOH, H, additional, KUBOTA, M, additional, OSAMURA, Y, additional, OGUNI, Masami, additional, SHINOHARA, Haruo, additional, HATTA, Toshihisa, additional, TANAKA, Osamu, additional, Ohmura, M., additional, Hosokawa, Y., additional, Hattori, T., additional, OHNISHI, Yoshiyuki, additional, OHTANI, Haruo, additional, NAGURA, Hiroshi, additional, OHTAWARA, Yoshihisa, additional, SUZUKI, Kazuo, additional, ASO, Yoshi, additional, Fujii, Toshiyuki, additional, Okuda, Minoru, additional, Takahashi, Hideki, additional, Morii, Sotokichi, additional, Oishi, Yuji, additional, OKADA, Teruhiko, additional, KOBAYASHI, Toshihiro, additional, and SEGUCHI, Harumichi, additional

Published
1992
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183. Relationship between drinking patterns and the risk of type 2 diabetes: the Kansai Healthcare Study.

Author

Kyoko Kogawa Sato, Hayashi, Tomoshige, Harita, Nobuko, Koh, Hideo, Maeda, Isseki, Endo, Ginji, Nakamura, Yoshiko, Kambe, Hiroshi, and Kiyotaki, Chiharu

Subjects
BLOOD sugar analysis, TYPE 2 diabetes diagnosis, TYPE 2 diabetes risk factors, COMPARATIVE studies, CONFIDENCE intervals, ALCOHOL drinking, LONGITUDINAL method, QUESTIONNAIRES, RESEARCH funding, STATISTICAL hypothesis testing, BODY mass index, PROPORTIONAL hazards models, DATA analysis software
Abstract

Background Moderate alcohol consumption is associated with a decreased risk of type 2 diabetes. However, the relationship between drinking patterns, such as the weekly frequency of alcohol consumption and the quantity per drinking day, and the incidence of type 2 diabetes has not been sufficiently addressed. Methods Study participants included 10 631 Japanese men aged 40e55 years without type 2 diabetes at entry. Type 2 diabetes was diagnosed if a fasting plasma glucose level was $7.0 mmol/l or if participants were taking diabetes medications. Data on alcohol consumption were obtained from questionnaires. Results During the 37 172 person-years of follow-up, we confirmed 878 cases of type 2 diabetes. Frequent alcohol consumption was associated with a low risk of type 2 diabetes. Compared to non-drinkers, the multiple-adjusted HR for those who drank 4e7 days weekly was 0.76 (95% CI, 0.63 to 0.92). To assess the association between drinking pattern and type 2 diabetes, we examined the joint association of the weekly frequency and the quantity per drinking day with type 2 diabetes. Men who consumed 0.1e2.0 or 2.1e4.0 US standard drinks per drinking day on 4e7 days weekly had a lower risk of type 2 diabetes (HR 0.74, 95% CI 0.58 to 0.95; HR 0.74, 95% CI 0.60 to 0.91, respectively) compared to non-drinkers. Conclusions More frequent alcohol consumption lowered the risk of type 2 diabetes. Light to moderate alcohol consumption per drinking day on 4e7 days weekly lowered the risk of type 2 diabetes compared to non-drinkers. [ABSTRACT FROM AUTHOR]

Published
2012
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184. EP3 receptors mediate PGE2-induced hypothalamic paraventricular nucleus excitation and sympathetic activation.

Author

Zhi-Hua Zhang, Yang Yu, Shun-Guang Wei, Nakamura, Yoshiko, Nakamura, Kazuhiro, and Felder, Robert B.

Subjects
PROSTAGLANDIN E1, PROSTANOIDS, MISOPROSTOL, HEART beat, HEART ventricles, DRUG administration, LABORATORY rats
Abstract

Prostaglandin E2 (PGE2), an important mediator of the inflammatory response, acts centrally to elicit sympathetic excitation. PGE2 acts on at least four E-class prostanoid (EP) receptors known as EP1, EP2, EP3, and EP4. Since PGE2 production within the brain is ubiquitous, the different functions of PGE2 depend on the expression of these prostanoid receptors in specific brain areas. The type(s) and location(s) of the EP receptors that mediate sympathetic responses to central PGE2 remain unknown. We examined this question using PGE2, the relatively selective EP receptor agonists misoprostol and sulprostone, and the available selective antagonists for EP1, EP3, and EP4. In urethane-anesthetized rats, intracerebroventricular (ICV) administration of PGE2, sulprostone or misoprostol increased renal sympathetic nerve activity, blood pressure, and heart rate. These responses were significantly reduced by ICV pretreatment with the EP3 receptor antagonist; the EP1 and EP4 receptor antagonists had little or no effect. ICV PGE2 or misoprostol increased the discharge of neurons in the hypothalamic paraventricular nucleus (PVN). ICV misoprostol increased the c-Fos immunoreactivity of PVN neurons, an effect that was substantially reduced by the EP3 receptor antagonist. Real-time PCR detected EP3 receptor mRNA in PVN, and immunohistochemical studies revealed sparsely distributed EP3 receptors localized in GABAergic terminals and on a few PVN neurons. Direct bilateral PVN microinjections of PGE2 or sulprostone elicited sympathoexcitatory responses that were significantly reduced by the EP3 receptor antagonist. These data suggest that EP3 receptors mediate the central excitatory effects of PGE2 on PVN neurons and sympathetic discharge. [ABSTRACT FROM AUTHOR]

Published
2011
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185. GENERAL SESSION

Author

Tagawa, M., primary, Murakoshi, M., additional, Honma, S., additional, Isobe, M., additional, Suzuki, M., additional, Ohsawa, Y., additional, TAKAHASHI, Hideki, additional, OISHI, Yuji, additional, MORII, Sotokichi, additional, TAKAMATSU, Tetsuro, additional, MINAMIKAWA, Tetsuhiro, additional, FUJITA, Setsuya, additional, TAKAYA, Kenichi, additional, TOSHIMORI, Kiyotaka, additional, ARAKI, Sanae, additional, TANII, Ichiro, additional, Oura, Chikayoshi, additional, TOYODA, Mitsuyasu, additional, EBIHARA, Yoshiro, additional, ITO, Hisao, additional, KITA, Shigeo, additional, KUDO, Motoshige, additional, TSUYAMA, Shinichiro, additional, IHIDA, Kaori, additional, KASHIO, Nobuyuki, additional, MURATA, Fusayoshi, additional, UEDA, Hiromi, additional, FUJIMORI, Osamu, additional, YAMADA, Kazuyori, additional, KITAMURA, Hirokazu, additional, Ueki, Tetsuo, additional, Moriyama, Nobuo, additional, Tajima, Atsushi, additional, Higashihara, Eiji, additional, Aso, Yoshio, additional, UENO, Masaki, additional, NAKAMURA, Yoshiko, additional, Ukimura, Akira, additional, Kotaka, Masahiro, additional, Koide, Hisashi, additional, Deguchi, Hirofumi, additional, Kitaura, Yasushi, additional, Kawamura, Keishiro, additional, Hirai, Kanji, additional, UTSUMI, Yasufumi, additional, IWAMASA, Teruo, additional, Utsunomiya, H., additional, Komatsu, N., additional, Tsutsumi, Y., additional, Watanabe, K., additional, WATABIKI, Toshimitsu, additional, TOKIYASI, Takuma, additional, YOSHIDA, Manabu, additional, ISHIDA, Noriaki, additional, OGAWA, Kazuo, additional, Watanabe, Hitoshi, additional, Taira, Atsushi, additional, Kimura, Kumiko, additional, Asano, Goro, additional, Onda, Masahiko, additional, WATANABE, Jun, additional, TANAKA, Junichi, additional, KANAMURA, Shinsuke, additional, WATANABE, Masahito, additional, SHIMADA, Masahisa, additional, Yabu, Michihiro, additional, Senda, Takao, additional, Fujita, Hisao, additional, YAGITA, Kazuhiro, additional, OKAMURA, Hitoshi, additional, YOKOYAMA, Chihiro, additional, TAGUCHI, Jun-ichi, additional, NAGATSU, Ikuko, additional, IBATA, Yasuhiko, additional, YAMADA, Harutaka, additional, MIYAI, Hironobu, additional, FUTENMA, Arao, additional, KATO, Katumi, additional, YAMADA, Kazuto, additional, HIGASHIYAMA, Hidetoshi, additional, MORI, Masahiko, additional, Yamamoto, Noboru, additional, YAMANDA, Takeshi, additional, USUDA, Nobuteru, additional, SHIMIZU, Tadahiro, additional, HANAI, Toru, additional, NAGATA, Tetsuji, additional, IIDA, Futoshi, additional, Yamashita, S., additional, Shiozawa, M., additional, Yasuda, K., additional, YASUDA, Kenjiro, additional, YAMASHITA, Shuji, additional, Shiozawa, Masahide, additional, YASUI, Hitoshi, additional, YOKOSE, Satoshi, additional, TAJIMA, Yoshifumi, additional, UTSUMI, Nobuo, additional, YOKOTA, Sadaki, additional, YURI, Kazunari, additional, KAWATA, Mitsuhiro, additional, ZHANG, S.X., additional, KOBAYASHI, T., additional, OKADA, T., additional, SAZ, GARCIA DEL E., additional, and SEGUCHI, H., additional

Published
1991
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188. Disruption of the glycolate dehydrogenase gene in the high-CO2-requiring mutant HCR89 of Chlamydomonas reinhardtii.

Author

Nakamura, Yoshiko, Kanakagiri, Saradadevi, Kyujung Van, Wei He, and Spalding, Martin H.

Subjects
*DEHYDROGENASES, *GENES, *CHLAMYDOMONAS reinhardtii, *CHLAMYDOMONAS, *ALGAE, *LACTATES, *MITOCHONDRIA, *PLANT photorespiration, *PHOTOSYNTHESIS, *GREEN algae, *ENZYMES
Abstract

One of the most notable contrasts between the photorespiratory pathway of higher plants and that of many of the green algae including Chlamydomonas reinhardtii lies in the enzymes that serve for oxidation of glycolate to glyoxylate. The gene disrupted by insertional mutagenesis in a high-CO2-requiring mutant, HCR89, of C. reinhardtii was determined to encode glycolate dehydrogenase (EC 1.1.99.14), which serves as the counterpart of glycolate oxidase (EC 1.1.3.15) in classical higher plant photorespiration. Neither glycolate nor D-lactate oxidation from the membrane fraction of HCR89 was detected. Excretion of over-accumulated glycolate into media due to the absence of glycolate dehydrogenase activity was observed for HCR89 under both high- and low-CO2 conditions. Chlamydomonas glycolate dehydrogenase, CrGDH, with a molecular mass of 118 851 Da, comprises a relatively hydrophobic N-terminal region, a FAD-containing domain homologous to the D subunit of the glycolate oxidase complex from Escherischia coli, and an iron–sulfur cluster containing domain homologous to the C subunit of anaerobic glycerol-3-phosphate dehydrogenase complex from Escherichia coli. The second Cys residue in the second iron–sulfur cluster motif of CrGDH is replaced by Asp, as CxxDxxCxxxCP, indicating the second iron–sulfur cluster coordinates most likely 3Fe–4S instead of 4Fe–4S. The membrane association of the glycolate dehydrogenase activity agrees with three predicted transmembrane regions on the iron–sulfur domain. [ABSTRACT FROM AUTHOR]

Published
2005
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190. Identification of Sympathetic Premotor Neurons in Medullary Raphe Regions Mediating Fever and Other Thermoregulatory Functions.

Author

Nakamura, Kazuhiro, Matsumura, Kiyoshi, Hübschle, Thomas, Nakamura, Yoshiko, Hioki, Hiroyuki, Fujiyama, Fumino, Boldogköi, Zsolt, König, Matthias, Thiel, Heinz-Jürgen, Gerstberger, Rüdiger, Kobayashi, Shigeo, and Kaneko, Takeshi

Subjects
FEVER, NEURONS, PROSTAGLANDINS, SYMPATHETIC nervous system, BODY temperature regulation
Abstract

Sympathetic premotor neurons directly control sympathetic preganglionic neurons (SPNs) in the intermediolateral cell column (IML) of the thoracic spinal cord, and many of these premotor neurons are localized in the medulla oblongata. The rostral ventrolateral medulla contains premotor neurons controlling the cardiovascular conditions, whereas rostral medullary raphe regions are a candidate source of sympathetic premotor neurons for thermoregulatory functions. Here, we show that these medullary raphe regions contain putative glutamatergic neurons and that these neurons directly control thermoregulatory SPNs. Neurons expressing vesicular glutamate transporter 3 (VGLUT3) were distributed in the rat medullary raphe regions, including the raphe magnus and rostral raphe pallidus nuclei, and mostly lacked serotonin immunoreactivity. These VGLUT3-positive neurons expressed Fos in response to cold exposure or to central administration of prostaglandin E2, a pyrogenic mediator. Transneuronal retrograde labeling after inoculation of pseudorabies virus into the interscapular brown adipose tissue (BAT) or the tail indicated that those VGLUT3-expressing medullary raphe neurons innervated these thermoregulatory effector organs multisynaptically through SPNs of specific thoracic segments, and microinjection of glutamate into the IML of the BAT-controlling segments produced BAT thermogenesis. An anterograde tracing study further showed a direct projection of those VGLUT3-expressing medullary raphe neurons to the dendrites of SPNs. Furthermore, intra-IML application of glutamate receptor antagonists blocked BAT thermogenesis triggered by disinhibition of the medullary raphe regions. The present results suggest that VGLUT3-expressing neurons in the medullary raphe regions constitute excitatory neurons that could be categorized as a novel group of sympathetic premotor neurons for thermoregulatory functions, including fever. [ABSTRACT FROM AUTHOR]

Published
2004
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194. Immunoelectron Microscopic Localization of C-Terminus of 43-kDa Dystrophin-Associated Glycoprotein in Normal Human Skeletal Myofibers.

Author

Wakayama, Yoshihiro, Jimi, Takahiro, Takeda, Atsushi, Shibuya, Seiji, Nakamura, Yoshiko, and Oniki, Hiroaki

Abstract

Ultrastructural localization of the C-termlnus of a 43-kDa dystrophin-associated glycoprotein (43-DAG) and its relation to dystrophin were investigated in normal human skeletal myofibers by using single and double immunogold labelling methods. The C-terminal end of 43-DAG antibody was localized ultrastructuraOy at the inside surface of muscle plasma membrane and the sarcoplasmic side of the plasma membrane invaginations. Double immunolabelling electron microscopy disclosed that signals of the C-terminal end of 43-DAG and dystrophin were often associated with each other and formed a doublet at the inside surface of plasma membranes in normal human skeletal myofibers. [ABSTRACT FROM PUBLISHER]

Published
1994

196. Effective Medical Examination for VDT Workers Using a New Testing Method of Ocular Alignment.

Author

Matsuoka, Yohtaro, Nakamura, Yoshiko, and Kobatake, Takashi

Abstract

To investigate the relationship between asthenopia of VDT workers and the ocular alignment, we measured far and near alignment of 407 subjects (74 VDT workers with asthenopia, 137 VDT workers without asthenopia, and 196 non-VDT workers) using Total Vision tester (VT-500). There was no relationship between the asthenopia and the records of far alignment test. On the other hand, the incedence of exophoria and convergence insufficiency type in near alignment test was significantly high in the group of VDT workers with asthenopia. From these results, we speculate that cases who have exophoria or show incomplete fusional response measured under the near testing distance (50 cm) tend to complain of asthenopia. [ABSTRACT FROM AUTHOR]

Published
1998
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197. Cloning and Analysis of cDNA Encoding Rat Bleomycin Hydrolase, a DNA-Binding Cysteine Protease.

Author

Takeda, Atsushi, Masuda, Yutaka, Yamamoto, Takako, Hirabayashi, Takahiro, Nakamura, Yoshiko, and Nakaya, Kazuyasu

Subjects
HYDROLASES, ANTISENSE DNA, BLEOMYCIN, CYSTEINE proteinases, AMINO acid sequence
Abstract

We isolated and characterized almost the entire cDNA encoding BLM hydrolase from rat spleen cDNA libraries. The cDNA encoded a polypeptide composed of 454 amino acids, that had a slightly larger molecular mass than that was previously estimated by SDS-PAGE for purified BLhl hydrolase subunit. Amino acid sequence alignments showed that rat BLM hydrolase is homologous to that of rabbit (94% identity of partial amino acid sequence), yeast cysteine protease (39% identity), and the pepC gene products of three bacteria (34–40% identity). In addition, it contained the three regions that are conserved in other cysteine proteases and thought to constitute the catalytic center. These results indicated that rat BLM hydrolase is a member of the papain superfamily of cysteine proteases. Sequencing revealed several putative sites phosphorylated by different types of protein kinases, but no signal sequence, transmembrane domain, N-linked glycosylation site or DNA-binding motif. The yeast homolog is a DNA-binding cysteine protease [Xu et al. (1994) J. Biol. Chem. 269,21177-21183. We demonstrated that rat BLM hydrolase also binds the singlestranded form of the Gal4 DNA-binding site oligonucleotide with high affinity compared with that of the double-stranded form. Northern blots revealed that the level of BLM hydrolase mRNA expression was very low in the rat skin, lung, and skeletal muscle. Furthermore, BLM hydrolase mRNA was ubiquitously expressed in the human cell lines, HeLa, SKG-IIIa, FL, KB, HEp-2, U373 GM, P3HR-1, Raji, THE-1, Jurkat, and Molt-4. These results suggested that BLM hydrolase plays important physiological roles, including the metabolism of antibiotics. [ABSTRACT FROM AUTHOR]

Published
1996
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198. Purification and Characterization of Bleomycin Hydrolase, Which Represents a New Family of Cysteine Proteases, from Rat Skin.

Author

Takeda, Atsushi, Higuchi, Dousei, Yamamoko, Takako, Nakamura, Yoshiko, Masuda, Yutaka, Hirabayashi, Takahiro, and Nakaya, Kazuyasu

Subjects
AMINO acids, BIOCHEMISTRY, PROTEIN analysis, CHROMATOGRAPHIC analysis, ANTI-infective agents
Abstract

Bleomycin (BLM) hydrolase, which hydrolyzes the carboxyamide bond in the β-amino-alanine moiety, was purified from newborn rat skin. The enzyme was purified 2,500-fold over the crude extract to apparent homogeneity in five steps in the presence of 2-mercap-toethanol: 45-65% ammonium sulfate fractionation, followed by chromatographies on Sephacryl S−200, DEAE-cellulofine, Phe-Superose, and Mono Q ion-exchange. The native enzyme had a molecular mass of 280 kDa according to gel filtration. The subunit molecular mass was estimated as 48 kDa by SDS-PAGE, indicating that the enzyme was comprised of six identical sub units. The amino acid sequence of its NH2 -terminus was determined to be acetyl-Met-Asn-Asn-Ala-Gly-Leu-Asn-Ser-Glu-Lys-, which was not found in the ami no acid sequence database. The optimum pH of the enzyme was 7.5 with pepleomycin (PLM). The Km, and Kmax values were 2.1 mM and 6.8 μmol.mg−1.h−1 for PLM, and 1.8 mM and 7.2 μmol.mg−1.h−1 for BLM-A2, respectively. The enzyme activity was inhibited by iodoacetic acid, N-ethylmaleinimide (NEM), and P-chloromercuribenzoic acid (pCMB) as well as divalent cations such as Cu2+, Cd2+, Hg2+, and Zn2+. It was effectively inhibited by a cysteine protease inhibitor E-64. However, cystatins A and C did not inhibit the activity. BLM hydrolase exhibited broad aminopeptidase substrate specificity towards aminoacyl-β-naphthylamides such as basic, neutral, and hydrophobic amino acid residues, as well as acidic residues. These results indicated that BLM hydrolase represents a new family of cysteine proteases. Western blotting and immunohistochemical analyses showed that BLM hydrolase is ubiquitous in various rat tissues but at low levels in lung and adult skin tissues, suggesting that this enzyme plays an important role in the metabolism of antibiotics. [ABSTRACT FROM AUTHOR]

Published
1996
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200. Onset of type 1 diabetes mellitus and heparin‐induced thrombocytopenia in a patient with Basedow's disease and idiopathic thrombocytopenic purpura: Novel combination as autoimmune polyglandular syndrome.

Author

Kinoshita, Tomoe, Kamei, Shinji, Nakamura, Yoshiko, Shimoda, Masashi, Anno, Takatoshi, Obata, Atsushi, Kimura, Tomohiko, Hirukawa, Hidenori, Tatsumi, Fuminori, Kohara, Kenji, Nakanishi, Shuhei, Mune, Tomoatsu, Kaku, Kohei, and Kaneto, Hideaki

Subjects
TYPE 2 diabetes, AUTOIMMUNE diseases, GRAVES' disease, IDIOPATHIC thrombocytopenic purpura, DRUG side effects
Published
2018
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