Faculty Profiles - Uchiyama Jumpei

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Characterization of Helicobacter pylori Bacteriophage KHP30 Reviewed

Jumpei Uchiyama, Hiroaki Takeuchi, Shin-ichiro Kato, Keiji Gamoh, Iyo Takemura-Uchiyama, Takako Ujihara, Masanori Daibata, Shigenobu Matsuzaki

APPLIED AND ENVIRONMENTAL MICROBIOLOGY 79 ( 10 ) 3176 - 3184 2013.5

Authorship：Lead author Language：English Publisher：AMER SOC MICROBIOLOGY

Helicobacter pylori inhabits the stomach mucosa and is a causative agent of stomach ulcer and cancer. In general, bacteriophages (phages) are strongly associated with bacterial evolution, including the development of pathogenicity. Several tailed phages have so far been reported in H. pylori. We have isolated an H. pylori phage, KHP30, and reported its genomic sequence. In this study, we examined the biological characteristics of phage KHP30. Phage KHP30 was found to be a spherical lipid-containing phage with a diameter of ca. 69 nm. Interestingly, it was stable from pH 2.5 to pH 10, suggesting that it is adapted to the highly acidic environment of the human stomach. Phage KHP30 multiplied on 63.6% of clinical H. pylori isolates. The latent period was ca. 140 min, shorter than the doubling time of H. pylori (ca. 180 min). The burst size was ca. 13, which was smaller than the burst sizes of other known tailed or spherical phages. Phage KHP30 seemed to be maintained as an episome in H. pylori strain NY43 cells, despite a predicted integrase gene in the KHP30 genomic sequence. Seven possible virion proteins of phage KHP30 were analyzed using N-terminal protein sequencing and mass spectrometry, and their genes were found to be located on its genomic DNA. The genomic organization of phage KHP30 differed from the genomic organizations in the known spherical phage families Corticoviridae and Tectiviridae. This evidence suggests that phage KHP30 is a new type of spherical phage that cannot be classified in any existing virus category.

DOI： 10.1128/AEM.03530-12

C型ボツリヌス毒素変換ファージのコアゲノムの探索

阪口義彦, 内山淳平, 小椋義俊, 山本由弥子, 鈴木智典, 松崎茂展, 林哲也, 小熊惠二

日本細菌学雑誌 68 ( 1 ) 178 - 178 2013.2

ピロリ菌バクテリオファージ(ファージ)KHP30の形態とゲノム解析

内山淳平, 竹内啓晃, 阪口義彦, 氏原隆子, 内山伊代, 大畑雅典, 松崎茂展

日本細菌学雑誌 68 ( 1 ) 224 - 224 2013.2

Bacteria Detection using Adsorption Protein of Tailed Phages

内山淳平, 内山(竹村) 伊代, 渡辺茂

Bio industry 30 ( 2 ) 47 - 53 2013.2

Synergistic bacteriolysis by bacteriophage $ϕ$EF24C endolysin ORF9 and lantibiotic nisin and its application to pulsed-field gel electrophoretic analysis of Enterococcus faecalis Reviewed

Takemura-Uchiyama, Iyo, Uchiyama, Jumpei, Satoh, Miho, Ujihara, Takako, Daibata, Masanori, Matsuzaki, Shigenobu

Annals of Microbiology 63 ( 3 ) 1209 - 1211 2013

Language：English Publisher：Springer

Genotyping by pulsed-field gel electrophoresis (PFGE) is recognized as one of the most useful methods for epidemiological studies of drug-resistant Enterococcus faecalis nosocomial outbreaks. As the bacteriolysis procedure is a critical step in PFGE, an accurate and reliable alternative bacteriolytic method to the conventional protocol would be useful. In this study, we examined the applicability of endolysin ORF9, derived from the E. faecalis phage φEF24C, and lantibiotic nisin in sample preparation for PFGE. The results show that the cooperative actions of nisin and ORF9 synergistically lysed E. faecalis, which was then amenable to PFGE analysis. © 2012 Springer-Verlag Berlin Heidelberg and the University of Milan.

DOI： 10.1007/s13213-012-0556-y

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新規ファージS6の分離とブドウ球菌における普遍形質導入の検討

内山淳平, 内山(竹村)伊代, 蒲生恵司, 阪口義彦, 三澤尚明, 大畑雅典, 松崎茂展

日本ブドウ球菌研究会プログラム・講演要旨集 58th 25 2013

P182 Bacterial detection system for Staphylococcus aureus based on bacteriophage tail adsorption protein Reviewed

Uchiyama, J, Uchiyama-Takemura, I, Watanabe, S, Sakaguchi, Y, Daibata, M, Matsuzaki, S

International Journal of Antimicrobial Agents 42 S100 - S100 2013

黄色ブドウ球菌性肺感染症に対するファージ療法の可能性の検討

松崎茂展, 内山淳平, 内山(竹村)伊代, 阪口義彦, 加藤伸一郎, 氏原隆子, 大畑雅典

日本ブドウ球菌研究会プログラム・講演要旨集 58th 26 2013

難治性黄色ブドウ球菌感染症に対するバクテリオファージ療法の開発―治療用ファージバンクの構築―

内山淳平, 竹村伊代, 氏原隆子, 松崎茂展, 大畑雅典

感染症学雑誌 86 ( 5 ) 637 2012.9

ファージリガンド分子を利用した細菌検査法の開発

内山淳平, 竹村伊代, 氏原隆子, 松崎茂展, 大畑雅典

感染症学雑誌 86 ( 4 ) 506 2012.7

Genetic characterization of Pseudomonas aeruginosa bacteriophage KPP10 Reviewed

Jumpei Uchiyama, Mohammad Rashel, Iyo Takemura, Shin-ichiro Kato, Takako Ujihara, Asako Muraoka, Shigenobu Matsuzaki, Masanori Daibata

ARCHIVES OF VIROLOGY 157 ( 4 ) 733 - 738 2012.4

Authorship：Lead author Language：English Publisher：SPRINGER WIEN

Bacteriophage (phage) KPP10 has been used in experimental phage therapies directed against P. aeruginosa infections. To examine the eligibility of phage KPP10 as a therapeutic phage, its genome was analyzed. The genomic DNA was shown to be 88,322 bp long, with 158 open reading frames (ORFs), and three tRNA genes were predicted. No ORF-encoded pathogenicity or lysogenization factor was predicted. A comparative genomic analysis revealed that phage KPP10, together with phage PAK_P3, can be grouped as a new type of lytic phage infecting P. aeruginosa. Phage KPP10 is considered to be suitable for therapeutic purposes because it is a lytic phage without ORF-encoded pathogenicity or a lysogenization factors.

DOI： 10.1007/s00705-011-1210-x

ファージリガンド分子を利用した細菌検査法の開発

内山淳平, 竹村伊代, 氏原隆子, 松崎茂展, 大畑雅典

感染症学雑誌 86 ( 臨増 ) 225 - 225 2012.3

難治性黄色ブドウ球菌感染症に対するバクテリオファージ療法の開発治療用ファージバンクの構築

内山淳平, 竹村伊代, 氏原隆子, 松崎茂展, 大畑雅典

感染症学雑誌 86 ( 臨増 ) 291 - 291 2012.3

Improved adsorption of an Enterococcus faecalis bacteriophage phiEF24C with a point mutation

UCHIYAMA Jumpei, TAKEMURA Iyo, UJIHARA Takako, MATSUZAKI Shigenobu, DAIBATA Masanori

日本細菌学雑誌 67 ( 1 ) 122 - 122 2012.2

点突然変異を有するEnterococcus faecalisバクテリオファージphiEF24Cの吸収改善(Improved adsorption of an Enterococcus faecalis bacteriophage phiEF24C with a point mutation)

内山淳平, 竹村伊代, 氏原隆子, 松崎茂展, 大畑雅典

日本細菌学雑誌 67 ( 1 ) 122 - 122 2012.2

Analysis of deoxynucleosides in bacteriophages $ϕ$EF24C and K and the frequency of a specific restriction site in the genomes of members of the bacteriophage subfamily Spounavirinae Reviewed

Uchiyama, Jumpei, Maeda, Yusuke, Takemura, Iyo, Gamoh, Keiji, Matsuzaki, Shigenobu, Daibata, Masanori

Archives of virology 157 ( 8 ) 1587 - 1592 2012

Language：English Publisher：Springer

The genera SPO1-like and Twort-like viruses in the subfamily Spounavirinae of the family Myoviridae have been newly proposed, with the reorganization of the SPO1-related bacteriophages (phages). A criterion defining these viral genera is the presence/absence of DNA modifications. In this study, liquid chromatography/mass spectrometry showed that phages I center dot EF24C and K of the subfamily Spounavirinae have unmodified DNA, which classifies them as Twort-like viruses. Moreover, in the subfamily Spounavirinae, DNA modification and elimination of a particular DNA sequence were suggested to be the major antirestriction strategies of the SPO1-like and Twort-like viruses, respectively.

DOI： 10.1007/s00705-012-1324-9

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C型ボツリヌス毒素変換ファージのゲノム情報を基盤とした偽溶原性メカニズムの解析

阪口義彦, OLIVA Maria A, MARTIN‐GALIANO Antonio J, ANDREU Jose M, 阪口政清, 内山淳平, 小椋義俊, 山本由弥子, 鈴木智典, 織田華絵, 津田千秋, 松崎茂展, 林哲也, 小熊惠二

日本分子生物学会年会プログラム・要旨集(Web) 35th WEB ONLY 3W3III-2 2012

緑膿菌臨床分離株に対するバクテリオファージKPP12の溶菌効果の検討

福田憲, 内山淳平, 森田珠恵, 石田わか, 角環, 松崎茂展, 大畑雅典, 福島敦樹

日本角膜学会総会・日本角膜移植学会プログラム・抄録集 36th-28th 45 2012

バクテリオファージ療法研究の現状と展望 Invited

松崎茂展, 内山淳平, 竹村伊代, 大畑雅典

週刊日本医事新報 4551 ( 4551 ) 48 - 49 2011.7

Urothelial/lamina propria spontaneous activity and the role of M3 muscarinic receptors in mediating rate responses to stretch and carbachol Reviewed

Moro, Christian, Uchiyama, Jumpei, Chess-Williams, Russ

Urology 78 ( 6 ) 1442 - e9 2011

Language：English Publisher：Elsevier

OBJECTIVE To investigate the effects of tissue stretch and muscarinic receptor stimulation on the spontaneous activity of the urothelium/ lamina propria and identify the specific receptor subtype mediating these responses.
METHODS Isolated strips of porcine urothelium with lamina propria were set up for in vitro recording of contractile activity. Muscarinic receptor subtype-selective antagonists were used to identify the receptors influencing the contractile rate responses to stretch and stimulation with carbachol.
RESULTS Isolated strips of urothelium with lamina propria developed spontaneous contractions (3.7 cycles/min) that were unaffected by tetrodotoxin, N omega-nitro-L-arginine, or indomethacin. Carbachol (1 mu M) increased the spontaneous contractile rate of these tissue strips by 122% +/- 27% (P < .001). These responses were significantly depressed in the presence of the M3-selective muscarinic antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (10-30 nM) but were not affected by the M1-selective antagonist pirenzepine (30-100 nM) or the M2-selective antagonist methoctramine (0.1-1 mu M). Stretching of the tissue also caused an increase in the spontaneous contractile rate, and these responses were abolished by atropine (1 mu M) and low concentrations of 4-diphenylacetoxy-N-methylpiperidine methiodide (10 nM). Darifenacin, oxybutynin, tolterodine, and solifenacin (1 mu M) all significantly depressed the frequency responses to carbachol (1 mu M).
CONCLUSION The urothelium with the lamina propria exhibits a spontaneous contractile activity that is increased during stretch. The mechanism appears to involve endogenous acetylcholine release acting on M3 muscarinic receptors. Anticholinergic drugs used clinically depress the responses of these tissues, and this mechanism might represent an additional site of action for these drugs in the treatment of bladder overactivity. UROLOGY 78: 1442. e9-1442. e15, 2011. (C) 2011 Elsevier Inc.

DOI： 10.1016/j.urology.2011.08.039

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