Updated on 2022/11/17

写真a

 
Uchiyama Jumpei
 
Organization
Faculty of Medicine, Dentistry and Pharmaceutical Sciences Associate Professor
Position
Associate Professor
External link

Degree

  • 博士(医学) ( 高知大学大学院 )

Research Interests

  • ウイルス

  • 細菌

  • 抗菌酵素

  • Microbiome

  • One Health

  • バクテリオファージ

  • 感染症

  • ファージ療法

  • アレルギー

  • 細菌叢

  • 迅速細菌検査法

Research Areas

  • Life Science / Genetics

  • Life Science / Bacteriology

  • Life Science / Infectious disease medicine

  • Life Science / Veterinary medical science

  • Life Science / Applied microbiology

Research History

  • 岡山大学学術研究院医歯薬学域   准教授

    2021.9

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  • 麻布大学獣医学部   准教授

    2021.4 - 2021.8

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  • Azabu University   School of Veterinary Medicine   Lecturer

    2015.4 - 2021.3

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  • Kochi University   医歯学系   Assistant Professor

    2009.12 - 2015.3

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  • 高知大学   短期研究員

    2009.4 - 2009.11

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  • Kochi University   Medical School

    2009.4 - 2009.11

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  • Faculty of Medicien, Bond University, Australia   Research Fellow

    2008.3 - 2008.10

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  • 高知大学大学院医学系研究科   Graduate student(doctorate program)

    2006.4 - 2009.3

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Professional Memberships

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Committee Memberships

  • 日本ワンヘルスサイエンス学会   評議員  

    2022.10   

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    Committee type:Academic society

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  •   Bacterial viruses subcommittee, International Committee on Taxonomy of Viruses,  

    2017.12   

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    Committee type:Academic society

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  •   Grant Reviewer, Science Fund of the Republic of Serbia  

    2021.1 - 2021.12   

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    Committee type:Government

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  •   Grant reviewer, Polish academy of sciences  

    2018   

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    Committee type:Government

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  • 日本ブドウ球菌研究会   実行委員  

    2015.4   

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Papers

  • Phylogenic analysis of new viral cluster of large phages with unusual DNA genomes containing uracil in place of thymine in gene-sharing network, using phages S6 and PBS1 and relevant uncultured phages derived from sewage metagenomics Reviewed

    Jumpei Uchiyama, Iyo Takemura-Uchiyama, Kazuyoshi Gotoh, Shin-ichiro Kato, Yoshihiko Sakaguchi, Hironobu Murakami, Tomoki Fukuyama, Mao Kaneki, Osamu Matsushita, Shigenobu Matsuzaki

    Virus Research   319   198881 - 198881   2022.10

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    Authorship:Lead author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    Bacteriophages (phages) are the most diverse and abundant life-form on Earth. Jumbophages are phages with double-stranded DNA genomes longer than 200 kbp. Among these, some jumbophages with uracil in place of thymine as a nucleic acid base, which we have tentatively termed “dU jumbophages” in this study, have been reported. Because the dU jumbophages are considered to be a living fossil from the RNA world, the evolutionary traits of dU jumbophages are of interest. In this study, we examined the phylogeny of dU jumbophages. First, tBLASTx analysis of newly sequenced dU jumbophages such as Bacillus phage PBS1 and previously isolated Staphylococcus phage S6 showed similarity to the other dU jumbophages. Second, we detected the two partial genome sequences of uncultured phages possibly relevant to dU jumbophages, scaffold_002 and scaffold_007, from wastewater metagenomics. Third, according to the gene-sharing network analysis, the dU jumbophages, including phages PBS1 and S6, and uncultured phage scaffold_002 formed a cluster, which suggested a new viral subfamily/family. Finally, analyses of the phylogenetic relationship with other phages showed that the dU jumbophage cluster, which had two clades of phages infecting Gram-negative and Gram-positive bacteria, diverged from the single ancestral phage. These findings together with previous reports may imply that dU jumbophages evolved from the same origin before divergence of Gram-negative and Gram-positive bacteria.

    DOI: 10.1016/j.virusres.2022.198881

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  • Characterization of the oral and fecal microbiota associated with atopic dermatitis in dogs selected from a purebred Shiba Inu colony. Reviewed International journal

    Jumpei Uchiyama, Takafumi Osumi, Keijiro Mizukami, Tomoki Fukuyama, Ayaka Shima, Asaka Unno, Iyo Takemura-Uchiyama, Yumi Une, Hironobu Murakami, Masahiro Sakaguchi

    Letters in Applied Microbiology   2022.9

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Atopic dermatitis (AD) is a chronic and relapsing multifactorial inflammatory skin disease that also affects dogs. The oral and gut microbiota are associated with many disorders, including allergy. Few studies have addressed the oral and gut microbiota in dogs, although the skin microbiota has been studied relatively well in these animals. Here, we studied the AD-associated oral and gut microbiota in 16 healthy and nine AD dogs from a purebred Shiba Inu colony. We found that the diversity of the oral microbiota was significantly different among the dogs, whereas no significant difference was observed in the gut microbiota. Moreover, a differential abundance analysis detected the Family_XIII_AD3011_group (Anaerovoracaceae) in the gut microbiota of AD dogs; however, no bacterial taxa were detected in the oral microbiota. Third, the comparison of the microbial co-occurrence patterns between AD and healthy dogs identified differential networks in which the bacteria in the oral microbiota that were most strongly associated with AD were related with human periodontitis, whereas those in the gut microbiota were related with dysbiosis and gut inflammation. These results suggest that AD can alter the oral and gut microbiota in dogs.

    DOI: 10.1111/lam.13828

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  • Acid-stable capsid structure of Helicobacter pylori bacteriophage KHP30 by single-particle cryoelectron microscopy. Reviewed International journal

    Ryosuke Kamiya, Jumpei Uchiyama, Shigenobu Matsuzaki, Kazuyoshi Murata, Kenji Iwasaki, Naoyuki Miyazaki

    Structure (London, England : 1993)   30 ( 2 )   300 - 312   2021.9

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    The acid-stable capsid structures of Helicobacter pylori phages KHP30 and KHP40 are solved at 2.7 and 3.0 Å resolutions by cryoelectron microscopy, respectively. The capsids have icosahedral T = 9 symmetry and consist of each 540 copies of 2 structural proteins, a major capsid protein, and a cement protein. The major capsid proteins form 12 pentagonal capsomeres occupying icosahedral vertexes and 80 hexagonal capsomeres located at icosahedral faces and edges. The major capsid protein has a unique protruding loop extending to the neighboring subunit that stabilizes hexagonal capsomeres. Furthermore, the capsid is decorated with trimeric cement proteins with a jelly roll motif. The cement protein trimer sits on the quasi-three-fold axis formed by three major capsid protein capsomeres, thereby enhancing the particle stability by connecting these capsomeres. Sequence and structure comparisons between the related Helicobacter pylori phages suggest a possible mechanism of phage adaptation to the human gastric environment.

    DOI: 10.1016/j.str.2021.09.001

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  • Use of Recombinant Endolysin to Improve Accuracy of Group B Streptococcus Tests. Reviewed International journal

    Hidehito Matsui, Jumpei Uchiyama, Masaya Ogata, Tadahiro Nasukawa, Iyo Takemura-Uchiyama, Shin-Ichiro Kato, Hironobu Murakami, Masato Higashide, Hideaki Hanaki

    Microbiology spectrum   9 ( 1 )   e0007721 - 11   2021.8

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Group B Streptococcus (GBS) causes serious neonatal infection via vertical transmission. The prenatal GBS screening test is performed at the late stage of pregnancy to avoid risks of infection. In this test, enrichment culture is performed, followed by GBS identification. Selective medium is used for the enrichment; however, Enterococcus faecalis, which is a potential contaminant in swab samples, can interfere with the growth of GBS. Such bacterial contamination can lead to false-negative results. Endolysin, a bacteriophage-derived enzyme, degrades peptidoglycan in the bacterial cell wall; it is a promising antimicrobial agent for selectively eliminating specific bacterial genera/species. In this study, we used the recombinant endolysin EG-LYS, which is specific to E. faecalis; the endolysin potentially enriched GBS in the selective culture. First, in the false-negative model (coculture of GBS and E. faecalis, which disabled GBS detection in the subsequent GBS identification test), EG-LYS treatment at 0.1 mg/ml improved GBS detection. Next, we used 548 vaginal swabs to test the efficacy of EG-LYS treatment in improving GBS detection. EG-LYS treatment (0.1 mg/ml) increased the GBS-positive ratio to 17.9%, compared to 15.7% in the control (phosphate-buffered saline [PBS] treatment). In addition, there were an increased number of GBS colonies under EG-LYS treatment in some samples. The results were supported by the microbiota analysis of the enriched cultures. In conclusion, EG-LYS treatment of the enrichment culture potentially improves the accuracy of the prenatal GBS screening test. IMPORTANCE Endolysin is a bacteriophage-derived enzyme that degrades the peptidoglycan in the cell wall of host bacteria; it could be used as an antimicrobial agent for selectively eliminating specific bacterial genera/species. Group B Streptococcus (GBS) causes neonatal infection via vertical transmission; prenatal GBS screening test, in which enrichment culture is followed by bacterial identification, is used to detect the presence of GBS in pregnant women. However, the presence of commensal bacteria such as Enterococcus faecalis in clinical specimens can inhibit GBS growth in the selective enrichment culture, resulting in false-negative result. Here, we demonstrated that the application of originally isolated endolysin in the enrichment culture improved the test accuracy by inhibiting unwanted E. faecalis growth and therefore avoiding false-negative results, not only in experimental settings, but also in tests using vaginal swabs.

    DOI: 10.1128/Spectrum.00077-21

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  • Analyses of propagation processes of Staphylococcus aureus bacteriophages S13' and S25-3 in two different taxonomies by definitive screening design. Reviewed International journal

    Ippei Takeuchi, Tadahiro Nasukawa, Ryosuke Sugimoto, Iyo Takemura-Uchiyama, Hironobu Murakami, Jumpei Uchiyama

    Virus research   298   198406 - 198406   2021.6

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier {BV}  

    To introduce phage therapy against multidrug-resistant Staphylococcus aureus in Western medicine, the establishment of phage manufacturing, particularly phage propagation, is indispensable. For the propagation of S. aureus phages, knowledge of the effects of phage types, process parameters, and analytical methodologies should be investigated. In this study, S. aureus phage propagations were studied in a flask with a new class of design of experiments, definitive screening design, using S. aureus phages S13' and S25-3 in different taxonomies. Four process parameters, namely, multiplicity of infection, bacterial density at infection, time of harvest, and temperature, were evaluated with the regression models based on the phage concentration data measured using plaque assay and quantitative polymerase chain reaction. As a result, phage propagations measured using plaque assay and quantitative polymerase chain reaction were overall similar to each other in the case of phage S13', while they differed in the case of phage S25-3. These results suggest that the propagation processes need to be developed according to phage type, and the choice of methodologies for phage concentration measurements should be carefully considered.

    DOI: 10.1016/j.virusres.2021.198406

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  • Examination of the fecal microbiota in dairy cows infected with bovine leukemia virus. Reviewed International journal

    Jumpei Uchiyama, Hironobu Murakami, Reiichiro Sato, Keijiro Mizukami, Takehito Suzuki, Ayaka Shima, Genki Ishihara, Kazuyuki Sogawa, Masahiro Sakaguchi

    Veterinary microbiology   240   108547 - 108547   2020.1

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER  

    Infection of cattle by bovine leukemia virus (BLV) causes significant economic losses in terms of milk and meat production in many countries. Because the gut microbiota may be altered by immunomodulation resulting from viral infections, we hypothesized that latent BLV infection would change the gut (i.e., rumen and hindgut) microbiota of infected cattle. In this study, we compared the gut microbiota of 22 uninfected and 29 BLV-infected Holstein-Friesian cows kept on the same farm, by 16S rRNA amplicon sequence analysis of fecal samples. First, we found that the fecal microbial diversity of BLV-infected cows differed slightly from that of uninfected cows. According to differential abundance analysis, some bacterial taxa associated with ruminal fermentation, such as Lachnospiraceae and Veillonellaceae families, were enriched in the fecal microbiota of uninfected cows. Second, the virus propagation ability of BLV strains was examined in vitro, and the correlation of the fecal microbiota with this virus propagation ability was analyzed. Higher virus propagation was shown to lead to less diversity in the microbiota. Differential abundance analysis showed that one bacterial taxon of genus Sanguibacteroides was negatively correlated with the virus propagation ability of BLV strains. Considering these results, BLV infection was speculated to decrease energy production efficiency in the cows via modification of rumen and hindgut microbiota, which partly relies on the virus propagation ability of BLV strains. This may explain the secondary negative effects of BLV infections such as increased susceptibility to other infections and decreased lifetime milk production and reproductive efficiency.

    DOI: 10.1016/j.vetmic.2019.108547

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  • Therapeutic Potential of an Endolysin Derived from Kayvirus S25-3 for Staphylococcal Impetigo. Reviewed International journal

    Ichiro Imanishi, Jumpei Uchiyama, Toshihiro Tsukui, Junzo Hisatsune, Kaori Ide, Shigenobu Matsuzaki, Motoyuki Sugai, Koji Nishifuji

    Viruses   11 ( 9 )   769 - 769   2019.8

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:{MDPI} {AG}  

    Impetigo is a contagious skin infection predominantly caused by Staphylococcus aureus. Decontamination of S. aureus from the skin is becoming more difficult because of the emergence of antibiotic-resistant strains. Bacteriophage endolysins are less likely to invoke resistance and can eliminate the target bacteria without disturbance of the normal microflora. In this study, we investigated the therapeutic potential of a recombinant endolysin derived from kayvirus S25-3 against staphylococcal impetigo in an experimental setting. First, the recombinant S25-3 endolysin required an incubation period of over 15 minutes to exhibit efficient bactericidal effects against S. aureus. Second, topical application of the recombinant S25-3 endolysin decreased the number of intraepidermal staphylococci and the size of pustules in an experimental mouse model of impetigo. Third, treatment with the recombinant S25-3 endolysin increased the diversity of the skin microbiota in the same mice. Finally, we revealed the genus-specific bacteriolytic effect of recombinant S25-3 endolysin against staphylococci, particularly S. aureus, among human skin commensal bacteria. Therefore, topical treatment with recombinant S25-3 endolysin can be a promising disease management procedure for staphylococcal impetigo by efficient bacteriolysis of S. aureus while improving the cutaneous bacterial microflora.

    DOI: 10.3390/v11090769

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  • Virus purification by CsCl density gradient using general centrifugation. Reviewed International journal

    Tadahiro Nasukawa, Jumpei Uchiyama, Satoshi Taharaguchi, Sumire Ota, Takako Ujihara, Shigenobu Matsuzaki, Hironobu Murakami, Keijirou Mizukami, Masahiro Sakaguchi

    Archives of virology   162 ( 11 )   3523 - 3528   2017.11

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer  

    Virus purification by cesium chloride (CsCl) density gradient, which generally requires an expensive ultracentrifuge, is an essential technique in virology. Here, we optimized virus purification by CsCl density gradient using general centrifugation (40,000 × g, 2 h, 4 °C), which showed almost the same purification ability as conventional CsCl density gradient ultracentrifugation (100,000 × g, 1 h, 4 °C) using phages S13' and φEF24C. Moreover, adenovirus strain JM1/1 was also successfully purified by this method. We suggest that general centrifugation can become a less costly alternative to ultracentrifugation for virus purification by CsCl densiy gradient and will thus encourage research in virology.

    DOI: 10.1007/s00705-017-3513-z

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  • Adsorption of Staphylococcus viruses S13 ' and S24-1 on Staphylococcus aureus strains with different glycosidic linkage patterns of wall teichoic acids Reviewed International journal

    Jumpei Uchiyama, Maya Taniguchi, Kenji Kurokawa, Iyo Takemura-Uchiyama, Takako Ujihara, Hidekatsu Shimakura, Yoshihiko Sakaguchi, Hironobu Murakami, Masahiro Sakaguchi, Shigenobu Matsuzaki

    JOURNAL OF GENERAL VIROLOGY   98 ( 8 )   2171 - 2180   2017.8

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:MICROBIOLOGY SOC  

    The group of phages belonging to the family Podoviridae, genus P68virus, including Staphylococcus viruses S1'' and S24-1, are important because of their benefits in phage therapy against Staphylococcus aureus infections. The O-glycosidic linkage patterns of wall teichoic acids (WTAs) in S. aureus cell walls seem to be important for adsorption of this phage group. In this study, the adsorption of Staphylococcus viruses S13' and S24-1 to S. aureus was examined using strains with modified WTA glycosidic linkage patterns. We found that the beta-O-N-acetylglucosamine of WTAs was essential for S13' adsorption, while N-acetylglucosamine, regardless of the alpha- and beta-O-glycosidic linkages of the WTAs, was essential for S24-1 adsorption. Next, examining the binding activities of their receptor-binding proteins (RBPs) to cell walls with different WTA glycosidic patterns, the beta-O-N-acetylglucosamine of the WTAs was essential for S13' RBP binding, while N-acetylglucosamine, regardless of the alpha- and b-O-glycosidic linkages of the WTAs, was essential for S24-1 RBP binding. Therefore, the results of the RBP binding assays were consistent with those of the phage adsorption assays. Bioinformatic analysis suggested that the RBPs of Staphylococcus viruses S13' and S24-1 were structurally similar to the RBPs of phage phi11 of the family Siphoviridae. Phylogenetic analysis of the RBPs indicated that two phylogenetic subclusters in the family Podoviridae were related to the glycosidic linkage patterns required for phage adsorption, possibly mediated by RBPs. We hope that this study will encourage the future development of therapeutic phages.

    DOI: 10.1099/jgv.0.000865

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  • Intragenus generalized transduction in Staphylococcus spp. by a novel giant phage Reviewed International journal

    Jumpei Uchiyama, Iyo Takemura-Uchiyama, Yoshihiko Sakaguchi, Keiji Gamoh, Shin-ichiro Kato, Masanori Daibata, Takako Ujihara, Naoaki Misawa, Shigenobu Matsuzaki

    ISME JOURNAL   8 ( 9 )   1949 - 1952   2014.9

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    Bacteriophage (phage)-mediated generalized transduction is expected to contribute to the emergence of drug-resistant staphylococcal clones in various environments. In this study, novel phage S6 was isolated from sewage and used to test generalized transduction in human-and animal-derived staphylococci. Phage S6 was a novel type of giant myophage, which possessed a DNA genome that contained uracil instead of thymine, and it could infect all of the tested staphylococcal species. The phage S6 appeared to be similar to the transducing phage PBS1, which infects Bacillus spp. Moreover, phage S6 facilitated the transduction of a plasmid in Staphylococcus aureus and from S. aureus to non-aureus staphylococcal species, as well as vice versa. Transduction of methicillin resistance also occurred in S. aureus. This is the first report of successful intragenus generalized transduction among staphylococci.

    DOI: 10.1038/ismej.2014.29

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  • Experimental phage therapy against lethal lung-derived septicemia caused by Staphylococcus aureus in mice Reviewed International journal

    Iyo Takemura-Uchiyama, Jumpei Uchiyama, Makoto Osanai, Norihito Morimoto, Tadashi Asagiri, Takako Ujihara, Masanori Daibata, Tetsuro Sugiura, Shigenobu Matsuzaki

    Microbes and Infection   16 ( 6 )   512 - 517   2014.6

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Nosocomial respiratory infections caused by methicillin-resistant Staphylococcus aureus (MRSA) can progress to lethal systemic infections. Bacteriophage (phage) therapy is expected to be effective against these critical infections. Previously, phage S13' was proposed as a potential therapeutic phage. We here examined phage treatment in a mouse model of lung-derived septicemia using phage S13'. Intraperitoneal phage administration at 6h postinfection reduced the severity of infection and rescued the infected mice. Phage S13' can efficiently lyse hospital-acquired MRSA strains causing pneumonia-associated bacteremia invitro. Thus, phage therapy may be a possible therapeutic intervention in staphylococcal lung-derived septicemia. © 2014 Institut Pasteur.

    DOI: 10.1016/j.micinf.2014.02.011

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  • Perspective: The age of the phage Reviewed International journal

    Shigenobu Matsuzaki, Jumpei Uchiyama, Iyo Takemura-Uchiyama, Masanori Daibata

    Nature   509 ( 7498 SUPPL. )   S9   2014.5

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    DOI: 10.1038/509S9a

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  • Antibiotic-resistant status and pathogenic clonal complex of canine Streptococcus canis-associated deep pyoderma. International journal

    Ichiro Imanishi, Keita Iyori, Akira Také, Ryota Asahina, Manami Tsunoi, Ryuji Hirano, Jumpei Uchiyama, Yoichi Toyoda, Yoshihiko Sakaguchi, Shunji Hayashi

    BMC veterinary research   18 ( 1 )   395 - 395   2022.11

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    BACKGROUND: Streptococcus canis causes deep pyoderma in canines, which raises concerns about the risk of isolates from lesions acquiring an antibiotic-resistant phenotype. It is necessary to identify effective antibiotics and the characteristics of the pathogenic cluster for S. canis-associated deep pyoderma. RESULTS: The signalment, molecular typing, and antibiotic-resistant status of S. canis isolated from deep pyoderma lesions (27 strains) and oral cavities (26 strains) were analyzed. Older dogs tended to have S. canis-associated deep pyoderma (15 of 27 dogs over 10 years old). Veterinarians chose quinolones for 10/16 cases (63%), even though the rate of quinolone-resistant strains of S. canis is 38-59%. Although 70% of the strains showed resistance to three or more antibiotic classes (37/53), 94% (50/53) strains showed sensitivity for penicillins. We also identified β-lactamase activity among penicillin-resistant strains of S. canis. Clonal complex 13 (CC13) was detected only in lesions and formed independent clusters in the phylogenetic tree. One strain of CC13 was resistant to the anti-methicillin-resistant Staphylococcus aureus drugs, vancomycin and linezolid. CONCLUSION: Although antibiotic-resistant strains of S. canis are isolated at a high rate, they can currently be treated with β-lactamase-inhibiting penicillins. CC13 may be a pathogenic cluster with high levels of antibiotics resistance.

    DOI: 10.1186/s12917-022-03482-3

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  • Effects of Inherent Lactic Acid Bacteria on Inhibition of Angiotensin I-Converting Enzyme and Antioxidant Activities in Dry-Cured Meat Products Reviewed

    Masaya Ogata, Jumpei Uchiyama, Abdulatef M. Ahhmed, Seiichi Sakuraoka, Satoshi Taharaguchi, Ryoichi Sakata, Wataru Mizunoya, Shiro Takeda

    Foods   11 ( 14 )   2123 - 2123   2022.7

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    Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    The aim of this study was to investigate the inherent bacteria that contribute to expressing the angiotensin I-converting enzyme (ACE) inhibitory activity and the antioxidant activity of dry-cured meat products without a bacterial starter. Among the ten dry-cured meat product samples, Coppa and Milano salami exhibited high ACE inhibitory activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, and oxygen radical absorbance capacity (ORAC). No consistent trend was observed in the pH values or the total peptide and imidazole dipeptide concentration of the products that exhibited high ACE inhibitory and antioxidant activities in the tested samples. To investigate the bacteria contributing to the ACE inhibitory and antioxidant activities of the product, 16S rRNA sequencing analysis, isolation, and identification of bacteria were performed using not only Coppa and Milano salami but also the Jamon Serrano and Parma prosciutto products that had low functional activities. Results suggest the Lactobacillales order, particularly the species Latilactobacillus sakei and Pediococcus pentosaceus, were the main inherent bacteria in Coppa and Milano salami, respectively, compared with the Jamon Serrano and Parma prosciutto products. Therefore, the inherent lactic acid bacteria in dry-cured meat products without bacterial starter is important for ACE inhibitory and antioxidant activities of the products.

    DOI: 10.3390/foods11142123

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  • Screening of bacterial DNA in bile sampled from healthy dogs and dogs suffering from liver- or gallbladder-associated disease. Reviewed

    Sakurako Neo, Iyo Takemura-Uchiyama, Jumpei Uchiyama, Hironobu Murakami, Ayaka Shima, Hideki Kayanuma, Taiki Yokoyama, Satoshi Takagi, Eiichi Kanai, Masaharu Hisasue

    The Journal of veterinary medical science   84 ( 7 )   1019 - 1022   2022.5

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    Authorship:Corresponding author   Language:English  

    Although the biliary system is generally aseptic, gallbladder microbiota has been reported in humans and some animals apart from dogs. We screened and analyzed the bacterial deoxyribonucleic acid in canine gallbladders using bile sampled from 7 healthy dogs and 52 dogs with liver- or gallbladder-associated disease. PCR screening detected bacteria in 17.3% of diseased dogs (9/52) and none in healthy dogs. Microbiota analysis of PCR-positive samples showed that the microbial diversity differed between liver- and gallbladder-associated disease groups. Thus, a specific bacterial community appears to occur at a certain frequency in the bile of diseased dogs.

    DOI: 10.1292/jvms.22-0090

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  • Intracameral Bacteriophage Injection as Postoperative Prophylaxis for Enterococcus faecalis-Induced Endophthalmitis After Cataract Surgery in Rabbits. Reviewed International journal

    Tatsuma Kishimoto, Waka Ishida, Isana Nakajima, Takako Ujihara, Takashi Suzuki, Jumpei Uchiyama, Shigenobu Matsuzaki, Ken Fukuda

    Translational vision science & technology   11 ( 4 )   2 - 2   2022.4

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    Purpose: Post-cataract surgery bacterial endophthalmitis is a serious postoperative complication, and Enterococcus spp.-induced endophthalmitis reportedly has a particularly poor visual prognosis. This study aimed to demonstrate the prophylactic effect of postoperative intracameral phage administration in Enterococcus faecalis-induced endophthalmitis after cataract surgery in rabbits. Methods: Endophthalmitis was induced in rabbits by injecting E. faecalis into the anterior chamber just after lensectomy while simultaneously administering either phage phiEF24C-P2 or vehicle. Retinal function was evaluated using electroretinography. The number of viable bacteria and myeloperoxidase (MPO) activity in the eye and histopathologic examinations were analyzed 48 hours after infection. Results: In the vehicle-treated group, retinal function at 24 hours after infection was impaired, and the number of viable bacteria and MPO activity in the eye increased 48 hours later. In the phage-administered group, retinal function was maintained; the number of viable bacteria and MPO activity were significantly suppressed. Histopathologic examinations showed disruption of the retinal layers and the presence of numerous E. faecalis in the lens capsule and vitreous cavity in vehicle-treated eyes. In contrast, retinal structures were intact, and no E. faecalis staining was observed in phage-treated eyes. No retinal dysfunction was observed in the group that received phage only without lensectomy; almost no phage was detected in the eyes after 14 days of treatment. Conclusions: Phage administration in the anterior chamber did not cause retinal dysfunction and suppressed postoperative endophthalmitis in rabbits. Translational Relevance: In vivo results of intracameral phage administration suggest that phages are a promising prophylactic candidate for postoperative endophthalmitis.

    DOI: 10.1167/tvst.11.4.2

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  • Heterogeneous IgE reactivities to Staphylococcus pseudintermedius strains in dogs with atopic dermatitis, and the identification of DM13-domain-containing protein as a bacterial IgE-reactive molecule. Reviewed International journal

    Iyo Takemura-Uchiyama, Hiroki Tsurui, Hidekatsu Shimakura, Tadahiro Nasukawa, Ichiro Imanishi, Jumpei Uchiyama, Tomoki Fukuyama, Shuji Sakamoto, Keiko Morisawa, Masato Fujimura, Hironobu Murakami, Shuji Kanamaru, Kenji Kurokawa, Keiko Kawamoto, Keita Iyori, Masahiro Sakaguchi

    FEMS microbiology letters   369 ( 1 )   2022.2

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    Staphylococcus pseudintermedius is one of the major pathogens causing canine skin infection. In canine atopic dermatitis (AD), heterogeneous strains of S. pseudintermedius reside on the affected skin site. Because an increase in specific IgE to this bacterium has been reported, S. pseudintermedius is likely to exacerbate the severity of canine AD. In this study, the IgE reactivities to various S. pseudintermedius strains and the IgE-reactive molecules of S. pseudintermedius were investigated. First, examining the IgE reactivities to eight strains of S. pseudintermedius using 141 sera of AD dogs, strain variation of S. pseudintermedius showed 10-63% of the IgE reactivities. This is different from the expected result based on the concept of Staphylococcus aureus clonality in AD patients. Moreover, according to the western blot analysis, there were more than four proteins reactive to IgE. Subsequently, the analysis of the common IgE-reactive protein at ∼15 kDa confirmed that the DM13-domain-containing protein was reactive in AD dogs, which is not coincident with any S. aureus IgE-reactive molecules. Considering these, S. pseudintermedius is likely to exacerbate AD severity in dogs, slightly different from the case of S. aureus in human AD.

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  • Anaphylaxis after vaccination for cats in Japan. Reviewed

    Megumi Yoshida, Keijiro Mizukami, Masaharu Hisasue, Ichiro Imanishi, Keigo Kurata, Masaki Ochiai, Masato Itoh, Tadahiro Nasukawa, Jumpei Uchiyama, Hajime Tsujimoto, Masahiro Sakaguchi

    The Journal of veterinary medical science   84 ( 1 )   149 - 152   2021.11

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    Severe adverse reactions in cats after vaccination were examined from 316 cases reported to the Ministry of Agriculture, Forestry and Fisheries (MAFF) in Japan during 15-year period from April 2004 to March 2019. We found that 130 (41%) showed anaphylaxis, and 99 (76%) of the 130 cases of anaphylaxis resulted in death. Veterinarians should be well prepared to deal with vaccine-associated anaphylaxis in cats. Bovine serum albumin (BSA) as indicator of purification was detected at high levels in commercially available feline vaccines. BSA might derive from fetal calf serum in culture media. This study provides useful information about anaphylaxis including critical details of the potential clinical signs associated with adverse events to feline vaccination.

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  • Bacterial Viruses Subcommittee and Archaeal Viruses Subcommittee of the ICTV: update of taxonomy changes in 2021. Reviewed International journal

    Mart Krupovic, Dann Turner, Vera Morozova, Mike Dyall-Smith, Hanna M Oksanen, Rob Edwards, Bas E Dutilh, Susan M Lehman, Alejandro Reyes, Diana P Baquero, Matthew B Sullivan, Jumpei Uchiyama, Jesca Nakavuma, Jakub Barylski, Mark J Young, Shishen Du, Poliane Alfenas-Zerbini, Alla Kushkina, Andrew M Kropinski, Ipek Kurtböke, J Rodney Brister, Cédric Lood, B L Sarkar, Tong Yigang, Ying Liu, Li Huang, Johannes Wittmann, Nina Chanishvili, Leonardo J van Zyl, Janis Rumnieks, Tomohiro Mochizuki, Matti Jalasvuori, Ramy K Aziz, Małgorzata Łobocka, Kenneth M Stedman, Andrey N Shkoporov, Annika Gillis, Xu Peng, François Enault, Petar Knezevic, Rob Lavigne, Sung-Keun Rhee, Virginija Cvirkaite-Krupovic, Cristina Moraru, Andrea I Moreno Switt, Minna M Poranen, Andrew Millard, David Prangishvili, Evelien M Adriaenssens

    Archives of virology   166 ( 11 )   3239 - 3244   2021.8

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    In this article, we - the Bacterial Viruses Subcommittee and the Archaeal Viruses Subcommittee of the International Committee on Taxonomy of Viruses (ICTV) - summarise the results of our activities for the period March 2020 - March 2021. We report the division of the former Bacterial and Archaeal Viruses Subcommittee in two separate Subcommittees, welcome new members, a new Subcommittee Chair and Vice Chair, and give an overview of the new taxa that were proposed in 2020, approved by the Executive Committee and ratified by vote in 2021. In particular, a new realm, three orders, 15 families, 31 subfamilies, 734 genera and 1845 species were newly created or redefined (moved/promoted).

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  • New dot-blot method for evaluating the effect of inactivators on mite and Japanese cedar pollen allergens. Reviewed International journal

    Megumi Yoshida, Keijiro Mizukami, Keigo Kurata, Tadahiro Nasukawa, Jumpei Uchiyama, Masahiro Sakaguchi

    Bioscience, biotechnology, and biochemistry   85 ( 10 )   2089 - 2092   2021.8

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    As a method of evaluating the effect of inactivaors on allergens while suppressing the effect of inactivator on the assay, we developed new dot-blot method which combines immunostaining and protein detection methods. This method is useful for evaluating whether the inactivator can inactivate allergens rather than removing them from the assay.

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  • IgE reactivity to milk components in dogs with cutaneous adverse food reactions. Reviewed

    Hidekatsu Shimakura, Tadahiro Nasukawa, Jumpei Uchiyama, Ryosuke Sugimoto, Ichiro Imanishi, Sumire Oota, Keijiro Mizukami, Masato Fujimura, Masahiro Sakaguchi

    The Journal of veterinary medical science   83 ( 10 )   1509 - 1512   2021.8

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    We investigated the IgE reactivity to crude and purified milk antigens in the sera of 112 dogs with cutaneous adverse food reactions (CAFRs). Of the 112 dogs, 33 (29%) had specific IgE for crude milk antigens. In the dogs with milk-specific IgE, IgE reactivity to casein, bovine serum albumin (BSA), α-lactalbumin, β-lactoglobulin, and bovine IgG were 81%, 85%, 39%, 27%, and 35%, respectively. Casein and BSA may be important allergens in dogs with CAFRs. Some canine vaccines contain casein hydrolysate as a stabilizer and the pooled serum with anti-casein IgE showed IgE reactivity to the vaccines containing it. Information about IgE reactivity to casein in dogs with CAFRs could be useful for predicting adverse reactions to the vaccines including casein hydrolysate.

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  • Anaphylaxis after rabies vaccination for dogs in Japan. Reviewed

    Megumi Yoshida, Keijiro Mizukami, Masaharu Hisasue, Ichiro Imanishi, Keigo Kurata, Masaki Ochiai, Masato Itoh, Tadahiro Nasukawa, Jumpei Uchiyama, Hajime Tsujimoto, Masahiro Sakaguchi

    The Journal of veterinary medical science   83 ( 8 )   1202 - 1205   2021.8

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    Severe adverse reactions after rabies vaccination in dogs were examined from 317 cases reported to the Ministry of Agriculture, Forestry and Fisheries (MAFF) in Japan during 15-year period from April 2004 to March 2019. We found that 109 of the 317 dogs showed anaphylaxis (0.15/100,000 vaccinated dogs), and 71 of the 109 cases of anaphylaxis resulted in death (0.10/100,000 vaccinated dogs). We measured bovine serum albumin (BSA) in four commercially available rabies vaccines and found the levels ranged from 0.1 to 16.6 µg/dose. Our survey showed that the rate of anaphylaxis to rabies vaccines in dogs is rare, although some cases of anaphylaxis resulted in death. Veterinarians should be well prepared to deal with vaccine-associated anaphylaxis.

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  • African pygmy hedgehog adenovirus: Virus replication, virus-induced cytopathogenesis and activation of mitogen-activated protein kinase signaling pathways in infected MDCK cells. Reviewed International journal

    Rongduo Wen, Hideharu Ochiai, Jumpei Uchiyama, Nanako Osawa, Mami Oba, Yukie Katayama, Kaixin Li, Tsutomu Omatsu, Kenichi Tamukai, Kaoru Suzuki, Hiroo Madarame, Shinji Makino, Tetsuya Mizutani

    Research in veterinary science   139   152 - 158   2021.7

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    We examined several aspects of African hedgehog adenovirus (AhAdv-1) that was isolated from an African pygmy hedgehog, including: replication kinetics of, virus-induced cytopathic effect (CPE), activation status of mitogen-activated protein kinase (MAPK) signaling pathways, and possible roles of these signaling pathways in virus replication and virus-induced CPE in MDCK cells. AhAdv-1 efficiently replicated and induced CPE in infected cells and caused accumulation of cleaved caspase-3 at 24 h post-infection (p.i.), suggesting apoptosis induction. Analysis of several intracellular signal transduction pathways, which are involved in apoptosis, showed activation of p38 MAPK, Akt and ERK1/2 pathways at 3 h p.i., and upregulation of phosphorylated SAPK/JNK at 24 h p.i. Although p38 MAPK inhibitor and SAPK/JNK inhibitor suppressed activation of the respective pathways in infected cells, they did not inhibit virus-induced CPE. Treatment of infected cells with inhibitor of the Akt pathway, the p38 pathway, the SAPK/JNK pathway or the ERK pathway revealed that inhibitors of p38 pathway inhibited viral replication by real-time PCR and TCID50 assay in infected MDCK cells, suggesting that AhAdv-1 uses p38 pathway for multiplication in infected cells.

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  • Specific antiviral effect of violaceoid E on bovine leukemia virus. Reviewed International journal

    Hironobu Murakami, Makoto Murakami-Kawai, Shinji Kamisuki, Shibasaki Hisanobu, Yukine Tsurukawa, Jumpei Uchiyama, Masahiro Sakaguchi, Kenji Tsukamoto

    Virology   562   1 - 8   2021.7

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    Bovine leukemia virus (BLV) infection has spread worldwide causing significant economic losses in the livestock industry. In countries with a high prevalence of BLV, minimizing economic losses is challenging; thus, research into various countermeasures is important for improving BLV control. Because anti-BLV drugs have not been developed, the present study explored a promising chemical compound with anti-BLV activity. Initially, screening of a chemical compound library revealed that violaceoid E (vioE), which is isolated from fungus, showed antiviral activity. Further analysis demonstrated that the antiviral effect of vioE inhibited transcriptional activation of BLV. Cellular thermal shift assay and pulldown assays provided evidence for a direct interaction between vioE and the viral transactivator protein, Tax. These data indicate that interference with Tax-dependent transcription could be a novel target for development of anti-BLV drugs. Therefore, it is suggested that vioE is a novel antiviral compound against BLV.

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  • In Vitro and In Vivo Evaluation of Three Newly Isolated Bacteriophage Candidates, phiEF7H, phiEF14H1, phiEF19G, for Treatment of Enterococcus faecalis Endophthalmitis. Reviewed International journal

    Tatsuma Kishimoto, Waka Ishida, Tadahiro Nasukawa, Takako Ujihara, Isana Nakajima, Takashi Suzuki, Jumpei Uchiyama, Daisuke Todokoro, Masanori Daibata, Atsuki Fukushima, Shigenobu Matsuzaki, Ken Fukuda

    Microorganisms   9 ( 2 )   1 - 12   2021.1

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    Post-operative endophthalmitis caused by Enterococcus spp. progresses rapidly and often results in substantial and irreversible vision loss. Therefore, novel alternative treatments that are effective against enterococcal endophthalmitis are required. Bacteriophage therapy has the potential to be an optional therapy for infectious diseases. Therefore, we investigated the therapeutic potential of three newly isolated enterococcal phages, phiEF7H, phiEF14H1, and phiEF19G, in E. faecalis-induced endophthalmitis. These phages could lyse the broad-range E. faecalis, including strains derived from endophthalmitis and vancomycin-resistant E. faecalis in vitro, as determined by the streak test. Morphological and genomic analyses revealed that these phages were classified into the Herelleviridae genus Kochikohdavirus. The whole genomes of these phages contained 143,399, 143,280, and 143,400 bp, respectively. Endophthalmitis was induced in mice by injection of three strains of E. faecalis derived from post-operative endophthalmitis or vancomycin-resistant strains into the vitreous body. The number of viable bacteria and infiltration of neutrophils in the eye were both decreased by intravitreous injection of phiEF7H, phiEF14H1, and phiEF19G 6 h after injection of all E. faecalis strains. Thus, these results suggest that these newly isolated phages may serve as promising candidates for phage therapy against endophthalmitis.

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  • Purification of membrane vesicles from Gram-positive bacteria using flow cytometry, after iodixanol density-gradient ultracentrifugation Reviewed International journal

    Tadahiro Nasukawa, Ryosuke Sugimoto, Jumpei Uchiyama, Iyo Takemura-Uchiyama, Hironobu Murakami, Ken Fukuda, Shigenobu Matsuzaki, Masahiro Sakaguchi

    Research in Microbiology   172 ( 1 )   2021.1

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    Membrane vesicles (MVs) play biologically important roles in Gram-positive bacteria, and purification is essential for their study. Although high-performance flow cytometry has the capability to quantify and isolate specific small particles, it has not been examined for MV isolation. In this study, we used high-performance flow cytometry to analyze MV from Gram-positive bacteria, Staphylococcus aureus and Bacillus subtilis, prepared by iodixanol density-gradient ultracentrifugation. Analysis of the quality of MV samples before and after sorting showed that the flow cytometric sorting provided higher purity and uniformity compared to gradient isolation alone. The MV purification method using flow cytometry should prove useful for applications requiring a very high purity of MV samples such as proteomic, metagenomic or lipidomic studies.

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  • Functionality of liquid smoke as an antimicrobial in cooked meat products: liquid smoke suppresses spoilage-related lactic acid bacteria Reviewed

    Shiro TAKEDA, Jumpei UCHIYAMA, Kazutoshi SUGITA, Hirofumi ENOMOTO, Abdulatef M AHHMED, Yuki KINOSHITA, Wataru MIZUNOYA, Yoshitaka ARIMA, Ryoichi SAKATA

    Food Science and Technology Research   27 ( 5 )   759 - 768   2021

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    Lactic acid bacteria (LAB) are known to promote deterioration of quality in meat products, such as discoloration and slime production. In this study, five types of liquid smoke, considered food additives, were assayed for potential anti-spoilage LABs from meat products. The spoilage LABs isolated from meat products were identified as Lactobacillus plantarum, Enterococcus malodoratus, Streptococcus thermophilus, or Carnobacterium maltaromaticum. The liquid smoke product (LS1), made from mixed wood, was more effective in terms of spoilage LAB isolate activities. It had the highest phenol content along with the most varied component among the tested products. In the model sausages containing 0.5 % LS1, which was incubated at 30 °C for 5 d for accelerating their deterioration, the enumeration of LAB and staphylococci was significantly lower than that of the control. Therefore, LS1 was suggested to be useful for suppressing not only LAB but also the other bacteria related to spoilage of meat products.

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  • Analysis of a plasmid encoding botulinum neurotoxin type G gene in Clostridium argentinense. Reviewed International journal

    Yoshihiko Sakaguchi, Jumpei Uchiyama, Akira Také, Kazuyoshi Gotoh, Masakiyo Sakaguchi, Tomonori Suzuki, Yumiko Yamamoto, Koji Hosomi, Tomoko Kohda, Masafumi Mukamoto, Shunji Kozaki, Shunji Hayashi, Keiji Oguma

    Anaerobe   66   102281 - 102281   2020.12

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    Clostridium argentinense produces botulinum neurotoxin type G (BoNT/G). We sequenced and analyzed the plasmid harboring the bont/G gene, designated pCAG, in C. argentinense strain 2740. The pCAG consisted of 140,070 bp containing the bont/G gene cluster. Although this gene cluster showed high similarities in its DNA sequence and ORF arrangement to those of other bont gene clusters, the other regions of the plasmid did not. A phylogenetic study suggested that pCAG had a unique evolutionary history compared with other clostridial bont-harboring plasmids. This suggests that pCAG is possibly a novel type of plasmid expressing the bont/G gene in C. argentinense.

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  • IgE reactivity to fish allergens from Pacific cod (Gadus macrocephalus) in atopic dogs. Reviewed International journal

    Ichiro Imanishi, Jumpei Uchiyama, Keijiro Mizukami, Junichi Kamiie, Keigo Kurata, Keita Iyori, Masato Fujimura, Kuniyoshi Shimakura, Koji Nishifuji, Masahiro Sakaguchi

    BMC veterinary research   16 ( 1 )   341 - 341   2020.9

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    BACKGROUND: IgE reactivity to fish allergens in atopic dogs, which are used as models for food allergy, has not been elucidated to date. We investigated IgE reactivity to crude extracts and purified allergens derived from the Pacific cod (Gadus macrocephalus) in atopic dogs to identify the allergenic proteins of cod. RESULTS: The levels of specific IgE to crude cod extracts were measured in the sera of 179 atopic dogs, including 27 dogs with cod allergy, using enzyme-linked immunosorbent assay (ELISA). Specific IgE to crude cod extracts were present in 36 (20%) of the 179 atopic dogs and in 12 (44%) of the 27 dogs with cod allergy. The allergens in crude cod extracts were analyzed by ELISA, immunoblotting, and liquid chromatography-tandem mass spectrometry. In allergen component analysis, IgE reactivity to tropomyosin and enolase was observed in the sera of dogs with cod allergy. IgE reactivity to parvalbumin, collagen, and tropomyosin was evaluated using the sera of atopic dogs that tested positive for specific IgE to crude cod extracts. Among the 36 dogs with IgE reactivity to crude cod extracts, 9 (25%), 14 (39%), and 18 (50%) dogs tested positive for specific IgE to parvalbumin, collagen, and tropomyosin, respectively. CONCLUSIONS: The IgE reactivity to cod allergens observed in dogs was similar to that in humans, and this finding further supports the use of atopic dogs with fish allergy as a model for fish allergy in humans.

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  • Development of multipurpose recombinant reporter bovine leukemia virus. Reviewed International journal

    Hironobu Murakami, Yusuke Yajima, Fumiaki Sato, Shinji Kamisuki, Satoshi Taharaguchi, Ken Onda, Sanggun Roh, Jumpei Uchiyama, Masahiro Sakaguchi, Kenji Tsukamoto

    Virology   548   226 - 235   2020.9

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    Bovine leukemia virus (BLV) is a global problem that results in significant economic losses to the livestock industry. We developed three virus strains by inserting the HiBiT reporter tag from NanoLuc luciferase (NLuc) into limited sites within BLV molecular clones. Initial analysis for site selection of the tag insertion revealed a permissible site immediately downstream of the viral envelope gene. Therefore, NLuc activity could be used to measure virus copy numbers in the supernatant and the levels of cell infection. Productivity and growth kinetics of the reporter virus were similar to those of the wild-type strain; therefore, the reporter virus can be used to characterize the replication of chimeric viruses as well as responses to the antiviral drug, amprenavir. Collectively, our results suggest that the BLV reporter virus with a HiBiT tag insertion is a highly versatile system for various purposes such as evaluating virus replication and antiviral drugs.

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  • IgE sensitivity to Malassezia pachydermatis and mite allergens in dogs with atopic dermatitis. Reviewed International journal

    Hironobu Ishimaru, Noriaki Okamoto, Masato Fujimura, Kazuki Miyaji, Hidekatsu Shimakura, Yukari Takase, Keijiro Mizukami, Jumpei Uchiyama, Douglas J DeBoer, Masahiro Sakaguchi

    Veterinary immunology and immunopathology   226   110070 - 110070   2020.8

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    In this study, dogs with atopic dermatitis were separated into non-food-induced atopic dermatitis (NFIAD) group (n = 15) and food-induced atopic dermatitis (FIAD) group (n = 37) based on an elimination diet test. IgE reactivity for crude Malassezia pachydermatis (M. pachydermatis) and house dust mites (HDM) allergen extracts was investigated in the two groups using fluorometric enzyme-linked immunosorbent assay (ELISA) and intradermal skin test (IDST). Nine (60%) of the 15 dogs in NFIAD group and 6 (16%) of the 37 dogs in FIAD group showed specific IgE for M. pachydermatis (Mann-Whitney U-test, P < 0.01). By immunoblotting analysis, the pooled serum samples from dogs with IgE for M. pachydermatis showed IgE reactivity for 50 kDa protein of M. pachydermatis. Twelve (80%) of the 15 dogs in NFIAD group and 8 (22%) of the 37 dogs in FIAD group showed specific IgE for HDM (Mann-Whitney U-test, P < 0.01). In addition, the dogs in NFIAD group significantly show a positive IDST to M. pachydermatis and HDM extracts compared with the dogs in FIAD group. The results suggest that dogs with NFIAD are at increased risk of becoming sensitized to the normal commensal organism M. pachydermatis compared with dogs with FIAD, perhaps co-sensitization occurred due to an HDM protease antigen's, Der f 1 and/or Der p 1, proteolytic activity related epidermal skin barrier defects. Treatment to limit skin colonization may thus be especially important in NFIAD.

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  • Taxonomy of prokaryotic viruses: 2018-2019 update from the ICTV Bacterial and Archaeal Viruses Subcommittee. Reviewed International journal

    Evelien M Adriaenssens, Matthew B Sullivan, Petar Knezevic, Leonardo J van Zyl, B L Sarkar, Bas E Dutilh, Poliane Alfenas-Zerbini, Małgorzata Łobocka, Yigang Tong, James Rodney Brister, Andrea I Moreno Switt, Jochen Klumpp, Ramy Karam Aziz, Jakub Barylski, Jumpei Uchiyama, Rob A Edwards, Andrew M Kropinski, Nicola K Petty, Martha R J Clokie, Alla I Kushkina, Vera V Morozova, Siobain Duffy, Annika Gillis, Janis Rumnieks, İpek Kurtböke, Nina Chanishvili, Lawrence Goodridge, Johannes Wittmann, Rob Lavigne, Ho Bin Jang, David Prangishvili, Francois Enault, Dann Turner, Minna M Poranen, Hanna M Oksanen, Mart Krupovic

    Archives of virology   165 ( 5 )   1253 - 1260   2020.5

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    This article is a summary of the activities of the ICTV's Bacterial and Archaeal Viruses Subcommittee for the years 2018 and 2019. Highlights include the creation of a new order, 10 families, 22 subfamilies, 424 genera and 964 species. Some of our concerns about the ICTV's ability to adjust to and incorporate new DNA- and protein-based taxonomic tools are discussed.

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  • Novel neuroprotective hydroquinones with a vinyl alkyne from the fungus, Pestalotiopsis microspora. Reviewed

    Kazuki Kanno, Yukine Tsurukawa, Shinji Kamisuki, Hisanobu Shibasaki, Keita Iguchi, Hironobu Murakami, Jumpei Uchiyama, Kouji Kuramochi

    The Journal of antibiotics   72 ( 11 )   793 - 799   2019.11

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    New hydroquinone derivatives bearing a vinyl alkyne, pestalotioquinols A and B, were isolated from a fungal culture broth of Pestalotiopsis microspora. The structures of these novel compounds were determined by interpretation of spectroscopic data (1D/2D NMR, MS, and IR), and the absolute configuration of the stereogenic center of pestalotioquinol A was assigned using the modified Mosher's method. Nerve growth factor-differentiated neuronal PC12 cells were pretreated with pestalotioquinols A and B and removed from the medium, and then treated with a generator of peroxynitrite (ONOO-), a reactive nitrogen species, to induce cell death. The cytotoxicity of the treated cells was assessed by measuring lactate dehydrogenase leakage. As a result, 1-3 μM pretreatment of pestalotioquinols A and B rescued neuronal PC12 cells from peroxynitrite-induced cytotoxicity and the protective activity was sustained after removing each compound from the medium. These results demonstrate that pestalotioquinol derivatives are a new class of hydroquinones possessing a vinyl alkyne and exhibiting relatively high neuroprotective effects.

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  • A point mutation to the long terminal repeat of bovine leukemia virus related to viral productivity and transmissibility. Reviewed International journal

    Hironobu Murakami, Haruna Todaka, Jumpei Uchiyama, Reiichiro Sato, Kazuyuki Sogawa, Masahiro Sakaguchi, Kenji Tsukamoto

    Virology   537   45 - 52   2019.11

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    It is important to establish the molecular basis of the high transmissibility of bovine leukemia virus (BLV) to develop new methods of preventing viral transmission. Hence, the aim of this study was to determine whether some strains had transmission advantages. First, we determined the whole BLV genome sequences of all 34 BLV-infected cows from one farm. Phylogenetic analysis divided strains into 26 major and 8 minor strains. The major strains dominantly spread independent of host factor, bovine leucocyte antigen. Further analysis, with molecular clones, associated transmissibility with viral productivity in vitro. In addition, the two groups could be classified by group-specific mutations. The reverse genetic approach demonstrated that a spontaneous mutation at nucleotide 175 of the BLV genome, which is located in the viral promoter region, could alter viral productivity by changing viral transactivation, suggesting that BLV transmissibility is affected by a spontaneous mutation associated with viral productivity.

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  • Therapeutic Effects of Intravitreously Administered Bacteriophage in a Mouse Model of Endophthalmitis Caused by Vancomycin-Sensitive or -Resistant Enterococcus faecalis. Reviewed International journal

    Tatsuma Kishimoto, Waka Ishida, Ken Fukuda, Isana Nakajima, Takashi Suzuki, Jumpei Uchiyama, Shigenobu Matsuzaki, Daisuke Todokoro, Masanori Daibata, Atsuki Fukushima

    Antimicrobial agents and chemotherapy   63 ( 11 )   2019.11

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    Endophthalmitis due to infection with Enterococcus spp. progresses rapidly and often results in substantial and irreversible vision loss. Given that the frequency of this condition caused by vancomycin-resistant Enterococcus faecalis has been increasing, the development of novel therapeutics is urgently required. We have demonstrated the therapeutic potential of bacteriophage ΦEF24C-P2 in a mouse model of endophthalmitis caused by vancomycin-sensitive (EF24) or vancomycin-resistant (VRE2) strains of E. faecalis Phage ΦEF24C-P2 induced rapid and pronounced bacterial lysis in turbidity reduction assays with EF24, VRE2, and clinical isolates derived from patients with E. faecalis-related postoperative endophthalmitis. Endophthalmitis was induced in mice by injection of EF24 or VRE2 (1 × 104 cells) into the vitreous. The number of viable bacteria in the eye increased to >1 × 107 CFU, and neutrophil infiltration into the eye was detected as an increase in myeloperoxidase activity at 24 h after infection. A clinical score based on loss of visibility of the fundus as well as the number of viable bacteria and the level of myeloperoxidase activity in the eye were all significantly decreased by intravitreous injection of ΦEF24C-P2 6 h after injection of EF24 or VRE2. Whereas histopathologic analysis revealed massive infiltration of inflammatory cells and retinal detachment in vehicle-treated eyes, the number of these cells was greatly reduced and retinal structural integrity was preserved in phage-treated eyes. Our results thus suggest that intravitreous phage therapy is a potential treatment for endophthalmitis caused by vancomycin-sensitive or -resistant strains of E. faecalis.

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  • Phage pharmacokinetics: Relationship with administration route Reviewed

    Shigenobu Matsuzaki, Jumpei Uchiyama

    Phage Therapy: A Practical Approach   43 - 57   2019.10

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  • Complete Genome Sequence of an Adenovirus-1 Isolate from an African Pygmy Hedgehog (Atelerix albiventris) Exhibiting Respiratory Symptoms in Japan. Reviewed International journal

    Hiroo Madarame, Jumpei Uchiyama, Kenichi Tamukai, Yukie Katayama, Nanako Osawa, Kaoru Suzuki, Tetsuya Mizutani, Hideharu Ochiai

    Microbiology resource announcements   8 ( 40 )   2019.10

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    This study reports the complete genome sequence of an African pygmy hedgehog adenovirus-1 isolate from an African pygmy hedgehog which displayed respiratory symptoms that included nasal discharge, sniffling, coughing, and respiratory distress. The viral genome is 31,764 bp long and shows four deletion sites compared to that of skunk adenovirus-1.

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  • Dark-Field Microscopic Detection of Bacteria using Bacteriophage-Immobilized SiO2@AuNP Core-Shell Nanoparticles. Reviewed International journal

    Masashi Imai, Kouhei Mine, Haruna Tomonari, Jumpei Uchiyama, Shigenobu Matuzaki, Yosuke Niko, Shingo Hadano, Shigeru Watanabe

    Analytical chemistry   91 ( 19 )   12352 - 12357   2019.10

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    To replace molecular biological and immunological methods, biosensors have recently been developed for the rapid and sensitive detection of bacteria. Among a wide variety of biological materials, bacteriophages have received increasing attention as promising alternatives to antibodies in biosensor applications. Thus, we herein present a rapid and highly selective detection method for pathogenic bacteria, which combines dark-field light scattering imaging with a plasmonic biosensor system. The plasmonic biosensor system employs bacteriophages as the biorecognition element and the aggregation-induced light scattering signal of gold nanoparticle-assembled silica nanospheres as a signal transducer. Using Staphylococcus aureus strain SA27 as a model analyte, we demonstrated that the plasmonic biosensor system detects S. aureus in the presence of excess Escherichia coli in a highly selective manner. After the sample and the S. aureus phage S13'-conjugated plasmon scattering probe were mixed, S. aureus detection was completed within 15-20 min with a detection limit of 8 × 104 colony forming units per milliliter.

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  • Therapeutic Effects of Intravitreously Administered Bacteriophage in a Mouse Model of Endophthalmitis Caused by Vancomycin-Sensitive or -Resistant Enterococcus faecalis Reviewed

    Jumpei Uchiyama

    Antimicrobial Agents and Chemotherapy   2019.8

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    <jats:p>Endophthalmitis due to infection with <jats:italic>Enterococcus</jats:italic> spp. progresses rapidly and often results in substantial and irreversible vision loss. Given that the frequency of this condition caused by vancomycin-resistant <jats:italic>Enterococcus faecalis</jats:italic> has been increasing, the development of novel therapeutics is urgently required. We have now demonstrated the therapeutic potential of bacteriophage ΦEF24C-P2 in a mouse model of endophthalmitis caused by vancomycin-sensitive (EF24) or vancomycin-resistant (VRE2) strains of <jats:italic>E. faecalis</jats:italic>. Phage ΦEF24C-P2 induced rapid and pronounced bacterial lysis in turbidity reduction assays with EF24, VRE2, and clinical isolates derived from patients with <jats:italic>E. faecalis</jats:italic>–related postoperative endophthalmitis. Endophthalmitis was induced in mice by injection of EF24 or VRE2 (1 × 10<jats:sup>4</jats:sup> cells) into the vitreous. The number of viable bacteria in the eye increased to &gt;1 × 10<jats:sup>7</jats:sup> colony forming units and neutrophil infiltration into the eye was detected as an increase in myeloperoxidase activity at 24 h after infection. A clinical score based on loss of visibility of the fundus as well as the number of viable bacteria and the level of myeloperoxidase activity in the eye were all significantly decreased by intravitreous injection of ΦEF24C-P2 6 h after injection of EF24 or VRE2. Whereas histopathologic analysis revealed massive infiltration of inflammatory cells and retinal detachment in vehicle-treated eyes, the number of these cells was greatly reduced and retinal structural integrity was preserved in phage-treated eyes. Our results thus suggest that intravitreous phage therapy is a potential treatment for endophthalmitis caused by vancomycin-sensitive or -resistant strains of <jats:italic>E. faecalis.</jats:italic></jats:p>

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  • Novel neuroprotective hydroquinones with a vinyl alkyne from the fungus, Pestalotiopsis microspora Reviewed

    Jumpei Uchiyama

    The Journal of Antibiotics   2019.7

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  • Therapeutic effects of intravitreous bacteriophage on Enterococcus faecalis endophthalmitis in mice Reviewed

    Fukuda Ken, Kishimoto Tatsuma, Ishida Waka, Suzuki Takashi, Uchiyama Jumpei, Matsuzaki Shigenobu, Daibata Masanori, Fukushima Atsuki

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE   60 ( 9 )   2019.7

  • Age-related analysis of the gut microbiome in a purebred dog colony Reviewed International journal

    Keijiro Mizukami, Jumpei Uchiyama, Hirotaka Igarashi, Hironobu Murakami, Takafumi Osumi, Ayaka Shima, Genki Ishiahra, Tadahiro Nasukawa, Yumi Une, Masahiro Sakaguchi

    FEMS Microbiology Letters   366 ( 8 )   2019.4

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    Dogs are model animals that can be used to study the gut microbiome. Although the gut microbiome is assumed to be closely related to aging, information pertaining to this relationship in dogs is limited. Here, we examined the association between the canine gut microbiome and age via a bacterial 16S rRNA gene amplicon sequence analysis in a colony of 43 Japanese purebred Shiba Inu dogs. We found that microbial diversity tended to decrease with aging. A differential abundance analysis showed an association of a single specific microbe with aging. The age-related coabundance network analysis showed that two microbial network modules were positively and negatively associated with aging, respectively. These results suggest that the dog gut microbiome is likely to vary with aging.

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  • IgE reactivity to Pacific cod (Gadus macrocephalus) fish allergens in dogs with canine atopic dermatitis Reviewed

    Imanishi Ichiro, Uchiyama Jumpei, Matsuda Takako, Mizukami Keijiro, Shimakura Hidekatsu, Nasukawa Tadahiro, Kamiie Junichi, Kurata Keigo, Fujimura Masato, Shimakura Kuniyoshi, Nishifuji Koji, Sakaguchi Masahiro

    JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY   143 ( 2 )   AB68 - AB68   2019.2

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  • Use of a Silkworm Larva Model in Phage Therapy Experiments Reviewed International journal

    Jumpei Uchiyama, Iyo Takemura-Uchiyama, Shigenobu Matsuzaki

    Methods in Molecular Biology   1898   173 - 181   2019

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    Antibiotic-resistant bacteria can cause intractable infections in humans and animals, with damaging effects to health care and economics. Phage therapy is considered a possible alternative to chemotherapy for treating infections, but still requires laborious in vivo experiments before its introduction into society and its further development. Recently, silkworm larvae have been recognized as highly convenient and useful model animals, and an alternative to higher animals. We describe the procedure for experimental phage therapy to treat Staphylococcus aureus infections in silkworm larvae.

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  • Potential Application of Bacteriophages in Enrichment Culture for Improved Prenatal Streptococcus agalactiae Screening Reviewed International journal

    Jumpei Uchiyama, Hidehito Matsui, Hironobu Murakami, Shin-ichiro Kato, Naoki Watanabe, Tadahiro Nasukawa, Keijiro Mizukami, Masaya Ogata, Masahiro Sakaguchi, Shigenobu Matsuzaki, Hideaki Hanaki

    VIRUSES-BASEL   10 ( 10 )   2018.10

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    Vertical transmission of Streptococcus agalactiae can cause neonatal infections. A culture test in the late stage of pregnancy is used to screen for the presence of maternal S. agalactiae for intrapartum antibiotic prophylaxis. For the test, a vaginal-rectal sample is recommended to be enriched, followed by bacterial identification. In some cases, Enterococcus faecalis overgrows in the enrichment culture. Consequently, the identification test yields false-negative results. Bacteriophages (phages) can be used as antimicrobial materials. Here, we explored the feasibility of using phages to minimize false-negative results in an experimental setting. Phage mixture was prepared using three phages that specifically infect E. faecalis: phiEF24C, phiEF17H, and phiM1EF22. The mixture inhibited the growth of 86.7% (26/30) of vaginal E. faecalis strains. The simple coculture of E. faecalis and S. agalactiae was used as an experimental enrichment model. Phage mixture treatment led to suppression of E. faecalis growth and facilitation of S. agalactiae growth. In addition, testing several sets of S. agalactiae and E. faecalis strains, the treatment with phage mixture in the enrichment improved S. agalactiae detection on chromogenic agar. Our results suggest that the phage mixture can be usefully employed in the S. agalactiae culture test to increase test accuracy.

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  • Piperacillin and ceftazidime produce the strongest synergistic phage-antibiotic effect in Pseudomonas aeruginosa Reviewed International journal

    Jumpei Uchiyama, Ryu Shigehisa, Tadahiro Nasukawa, Keijiro Mizukami, Iyo Takemura-Uchiyama, Takako Ujihara, Hironobu Murakami, Ichiro Imanishi, Koji Nishifuji, Masahiro Sakaguchi, Shigenobu Matsuzaki

    ARCHIVES OF VIROLOGY   163 ( 7 )   1941 - 1948   2018.7

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    The combined use of phage and antibiotics can show synergistic antimicrobial effects, so-called phage-antibiotic synergy (PAS). Here, we screened and examined PAS against Pseudomonas aeruginosa in vitro. Testing four different phages infecting P. aeruginosa, phage KPP22 classified within the family Myoviridae genus Pbunavirus showed PAS with the widest range of antibiotics, and showed PAS with anti-Pseudomonas drugs such as piperacillin and ceftazidime. Thus, evidence suggests that the combined use of phage and antibiotics is a promising therapeutic strategy against P. aeruginosa infections, with consideration needed regarding the optimal selection and adequate application timing of these phages and antibiotics.

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  • Recovery of mycobacteriophages from archival stocks stored for approximately 50 years in Japan Reviewed International journal

    Takako Ujihara, Jumpei Uchiyama, Tadahiro Nasukawa, Hiroki Ando, Hironobu Murakami, Naoya Ohara, Midori Ogawa, Toshio Yamazaki, Masanori Daibata, Masahiro Sakaguchi, Shigenobu Matsuzaki

    ARCHIVES OF VIROLOGY   163 ( 7 )   1915 - 1919   2018.7

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    Mycobacteriophage archival stocks have been kept for ca. 20-50 years in Japan. In this study, we attempted to recover mycobacteriophages from 50 archival stocks and briefly analyzed the recovered phages. The phages were recovered from 72.2% (13/18) of the lyophilized stocks that had been stored for 47-56 years. Moreover, the analysis of 12 representative recovered phages led to their classification as belonging to the family Siphoviridae, and seven of them were typed by polymerase chain reaction (PCR) targeting the gene that encodes the tape measure protein. Considering these results, lyophilization seems to be suitable for phage archival storage.

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  • Polymorphisms in the Helicobacter pylori NY43 strain and its prophage-cured derivatives Reviewed International journal

    Jumpei Uchiyama

    Microbiology   164 ( 6 )   877 - 882   2018.6

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    This study aimed to determine the characteristics of the Helicobacter pylori host NY43 strain and its prophage-cured derivative. H. pylori colonizing the human stomach cause many diseases. They show high genetic diversity, allowing the development of mutant strains that can form bacterial communities adapted to specific environmental conditions. Bacteriophage activities are associated with bacterial evolution, including pathogenicity development. Herein, we reported the complete genome sequence and genomic organization of two H. pylori prophages, KHP30 and KHP40; the effects of KHP30 on the behaviours of NY43 are not yet known. We showed that approximately 57 % prophage-cured derivatives spontaneously appeared in the exponential phase during liquid culture, and the biological characteristics of these derivatives differed from those of the host NY43. KHP30 reinfected the cured derivatives, and the curing ratio was influenced by culture conditions. KHP30 was shown to promote the development of a flexible H. pylori community with variable characteristics.

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  • Genome Sequences of 12 Mycobacteriophages Recovered from Archival Stocks in Japan Reviewed International journal

    Jumpei Uchiyama, Keijiro Mizukami, Koji Yahara, Shin-ichiro Kato, Hironobu Murakami, Tadahiro Nasukawa, Naoya Ohara, Midori Ogawa, Toshio Yamazaki, Shigenobu Matsuzaki, Masahiro Sakaguchi

    MICROBIOLOGY RESOURCE ANNOUNCEMENTS   6 ( 25 )   2018.6

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    Using Mycobacterium smegmatis mc2155, 12 siphoviruses were recovered from long-term archival stocks stored in Japan. Their genome sequences were 46.0 to 61.3 kbp with 63 to 68% G+C contents, which allowed them to be categorized within cluster W and subclusters A1, A2, B3, A7, I1, and K4.

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  • Taxonomy of prokaryotic viruses: 2017 update from the ICTV Bacterial and Archaeal Viruses Subcommittee Reviewed International journal

    Evelien M. Adriaenssens, Johannes Wittmann, Jens H. Kuhn, Dann Turner, Matthew B. Sullivan, Bas E. Dutilh, Ho Bin Jang, Leonardo J. van Zyl, Jochen Klumpp, Malgorzata Lobocka, Andrea I. Moreno Switt, Janis Rumnieks, Robert A. Edwards, Jumpei Uchiyama, Poliane Alfenas-Zerbini, Nicola K. Petty, Andrew M. Kropinski, Jakub Barylski, Annika Gillis, Martha R.C. Clokie, David Prangishvili, Rob Lavigne, Ramy Karam Aziz, Siobain Duffy, Mart Krupovic, Minna M. Poranen, Petar Knezevic, Francois Enault, Yigang Tong, Hanna M. Oksanen, J. Rodney Brister

    Archives of Virology   163 ( 4 )   1125 - 1129   2018.4

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  • Isolation of Bacteriophages for Fastidious Bacteria Reviewed International journal

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

    Bacteriophage Therapy   1693   3 - 10   2018

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    © Springer Science+Business Media LLC 2018. One of the most important factors for successful bacteriophage therapy is, undoubtedly, the isolation of excellent therapeutic candidate bacteriophages. There are only a few reports about active bacteriophages in the fastidious bacteria Helicobacter pylori. In this chapter, we describe a method for isolating and purifying KHP30-like bacteriophages in H. pylori, which have lytic and pseudolysogenic life cycles.

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  • Bovine leukemia virus G4 enhances virus production. Reviewed International journal

    Hironobu Murakami, Shotaro Asano, Jumpei Uchiyama, Reiichiro Sato, Masahiro Sakaguchi, Kenji Tsukamoto

    Virus research   238   213 - 217   2017.6

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    The nonstructural G4 gene of bovine leukemia virus (BLV) has been thought to function in virus replication. However, the discovery of the AS1 gene on the antisense strand of the G4 gene has affected this interpretation. In this study, we investigated the function of G4 in virus production independent of the AS1 gene using a reverse genetic approach, and briefly examined the association of the G4 protein with Tax, which is also a nonstructural protein that promotes virus replication. First, we constructed a mutant molecular clone of BLV with a nonsense mutation in G4 that had a minimal effect on the AS1 gene. Comparison of the wild-type and mutant molecular clones indicated that the nonsense mutation resulted in a reduction of virus in the culture supernatant and accumulation of viral RNA (vRNA) in cells. Moreover, G4 and Tax expression in cells was shown to synergistically enhance virus production. Therefore, we suggest that G4 enhances virus production through abrogation of vRNA accumulation.

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  • Bovine leukemia virus G4 enhances virus production Reviewed

    Hironobu Murakami, Shotaro Asano, Jumpei Uchiyama, Reiichiro Sato, Masahiro Sakaguchi, Kenji Tsukamoto

    Virus Research   238   213 - 217   2017.6

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    © 2017 Elsevier B.V. The nonstructural G4 gene of bovine leukemia virus (BLV) has been thought to function in virus replication. However, the discovery of the AS1 gene on the antisense strand of the G4 gene has affected this interpretation. In this study, we investigated the function of G4 in virus production independent of the AS1 gene using a reverse genetic approach, and briefly examined the association of the G4 protein with Tax, which is also a nonstructural protein that promotes virus replication. First, we constructed a mutant molecular clone of BLV with a nonsense mutation in G4 that had a minimal effect on the AS1 gene. Comparison of the wild-type and mutant molecular clones indicated that the nonsense mutation resulted in a reduction of virus in the culture supernatant and accumulation of viral RNA (vRNA) in cells. Moreover, G4 and Tax expression in cells was shown to synergistically enhance virus production. Therefore, we suggest that G4 enhances virus production through abrogation of vRNA accumulation.

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  • Inefficient viral replication of bovine leukemia virus induced by spontaneous deletion mutation in the G4 gene Reviewed International journal

    Hironobu Murakami, Jumpei Uchiyama, Sae Nikaido, Reiichiro Sato, Masahiro Sakaguchi, Kenji Tsukamoto

    Journal of General Virology   97 ( 10 )   2753 - 2762   2016.10

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    © 2016 The Authors. Enzootic bovine leucosis is caused by bovine leukemia virus (BLV) infection, which is highly prevalent in several regions of the world and significantly impacts the livestock industry. In BLV infection, the proviral load in the blood reflects disease progression. Although the BLV genome is highly conserved among retroviruses, genetic variation has been reported. However, the relationship between proviral load and genetic variation is poorly understood. In this study, we investigated the changes in proviral load in BLV-infected cattle in Japan and then identified and analysed a BLV strain pvAF967 that had a static proviral load. First, examining the proviral load in the aleukaemic cattle in 2014 and 2015, cow AF967 showed a static proviral load, while the other cows showed significant increases in proviral load. Sequencing the provirus in cow AF967 showed a deletion of 12 nt located in the G4 gene. An in vitro assay system using BLV molecular clone was set up to evaluate viral replication and production. In this in vitro assay, the deletion mutation in the G4 gene resulted in a significant decrease in viral replication and production. In addition, we showed that the deletion mutation did not affect the viral transcriptional activity of Tax protein, which is also important for virus replication. The emergence of strain pvAF967 that showed a static proviral load, combined with other retrovirus evolutionary traits, suggests that some BLV strains may have evolved to be symbiotic with cattle.

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  • IgE reactivity to hen egg white allergens in dogs with cutaneous adverse food reactions Reviewed International journal

    Hidekatsu Shimakura, Jumpei Uchiyama, Taku Saito, Kazuki Miyaji, Masato Fujimura, Kenichi Masuda, Noriaki Okamoto, Douglas J. DeBoer, Masahiro Sakaguchi

    Veterinary Immunology and Immunopathology   177   52 - 57   2016.9

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    © 2016 Elsevier B.V. Dogs with cutaneous adverse food reactions (CAFR) often have specific IgE to food allergens. Egg white, which is majorly composed of ovomucoid, ovalbumin, ovotransferrin, and lysozyme, is a food allergen in dogs. Information of the IgE reactivity to purified egg white allergens supports accurate diagnosis and efficiency treatment in humans. However, to the best of our knowledge, there have been no studies on the IgE reactivity to purified egg white allergens in dogs. Here, we investigated the IgE reactivity to crude and purified allergens of hen egg white in dogs with CAFR. First, when we examined serum samples from 82 dogs with CAFR for specific IgE to crude egg white by ELISA, 9.8% (8/82) of the dogs with CAFR showed the IgE reactivity to crude egg white. We then used sera from the eight dogs with positive IgE reactivity to crude egg white to examine the IgE reactivity to four purified allergens, ovomucoid, ovalbumin, ovotransferrin, and lysozyme, by ELISA. We found that 75% (6/8) of the dogs showed IgE reactivity to both ovomucoid and ovalbumin, and that 37.5% (3/8) of the dogs showed IgE reactivity to ovotransferrin. None (0/8) showed IgE reactivity to lysozyme. Moreover, validating these results, the immunoblot analyses were performed using the sera of the three dogs showing the highest IgE reactivity to crude egg white. Both anti-ovomucoid and anti-ovalbumin IgE were detected in the sera of these dogs, while anti-ovotransferrin IgE was not detected. Considering these, ovomucoid and ovalbumin appears to be the major egg white allergens in dogs with CAFR.

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  • Analyses of Short-Term Antagonistic Evolution of Pseudomonas aeruginosa Strain PAO1 and Phage KPP22 (Myoviridae Family, PB1-Like Virus Genus) Reviewed International journal

    Jumpei Uchiyama, Masato Suzuki, Koji Nishifuji, Shin-ichiro Kato, Reina Miyata, Tadahiro Nasukawa, Kotoe Yamaguchi, Iyo Takemura-Uchiyama, Takako Ujihara, Hidekatsu Shimakura, Hironobu Murakami, Noriaki Okamoto, Yoshihiko Sakaguchi, Keigo Shibayama, Masahiro Sakaguchi, Shigenobu Matsuzaki

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY   82 ( 15 )   4482 - 4491   2016.8

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    Pseudomonas aeruginosa causes serious intractable infections in humans and animals. Bacteriophage ( phage) therapy has been applied to treat P. aeruginosa infections, and phages belonging to the PB1-like virus genus in the Myoviridae family have been used as therapeutic phages. To achieve safer and more effective phage therapy, the use of preadapted phages is proposed. To understand in detail such phage preadaptation, the short-term antagonistic evolution of bacteria and phages should be studied. In this study, the short-term antagonistic evolution of bacteria and PB1-like phage was examined by studying phage-resistant clones of P. aeruginosa strain PAO1 and mutant PB1-like phages that had recovered their infectivity. First, phage KPP22 was isolated and characterized; it was classified as belonging to the PB1-like virus genus in the Myoviridae family. Subsequently, three KPP22-resistant PAO1 clones and three KPP22 mutant phages capable of infecting these clones were isolated in three sets of in vitro experiments. It was shown that the bacterial resistance to phage KPP22 was caused by significant decreases in phage adsorption and that the improved infectivity of KPP22 mutant phages was caused by significant increases in phage adsorption. The KPP22-resistant PAO1 clones and the KPP22 mutant phages were then analyzed genetically. All three KPP22-resistant PAO1 clones, which were deficient for the O5 antigen, had a common nonsense mutation in the wzy gene. All the KPP22 mutant phage genomes showed the same four missense mutations in the open reading frames orf060, orf065, and orf086. The information obtained in this study should be useful for further development of safe and efficient phage therapy.IMPORTANCEPseudomonas aeruginosa causes serious intractable infections in humans and animals; bacteriophage ( phage) therapy has been utilized to treat P. aeruginosa infections, and phages that belong to the PB1-like virus genus in the family Myoviridae have been used as therapeutic phages. The preadapted phage is trained in advance through the antagonistic evolution of bacteria and phage and is proposed to be used to achieve safer and more effective phage therapy. In this study, to understand the phage preadaptation, the in vitro short-term antagonistic evolution was studied using P. aeruginosa strain PAO1 and the newly isolated PB1-like phage KPP22. Phage KPP22 was characterized, and the molecular framework regarding the phage preadaptation of KPP22 was elucidated. The importance of study of antagonistic evolution of bacteria and phage in phage therapy is discussed.

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  • Screening of KHP30-like prophages among Japanese Helicobacter pylori strains, and genetic analysis of a defective KHP30-like prophage sequence integrated in the genome of the H. pylori strain NY40 Reviewed International journal

    Jumpei Uchiyama, Iyo Takemura-Uchiyama, Shin-ichiro Kato, Hiroaki Takeuchi, Yoshihiko Sakaguchi, Takako Ujihara, Masanori Daibata, Hidekatsu Shimakura, Noriaki Okamoto, Masahiro Sakaguchi, Shigenobu Matsuzaki

    FEMS MICROBIOLOGY LETTERS   363 ( 16 )   2016.8

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    We have recently reported the active Helicobacter pylori bacteriophages (phages), KHP30 and KHP40, the genomic DNAs of which exist as episomes in host bacterial strains isolated in Japan (i.e. pseudolysogeny). In this study, we examined the possibility of the lysogeny of active KHP30-like phages in Japanese H. pylori strains, because their genomes contain a putative integrase gene. Only the NY40 strain yielded partial detection of a KHP30-like prophage sequence in PCR among 174 Japanese H. pylori isolates, except for strains producing the above active phages. Next, according to the genomic analysis of the NY40 strain, the KHP30-like prophage sequence was found to be located from ca. 524 to 549 kb in the host chromosome. The attachment sites, attL and attR, in the NY40 genome showed almost the same genomic location and sequence as those detected in a French isolate B38, suggesting that an active parental KHP30-like phage had integrated into the ancestral NY40 genome in a site-specific manner. The prophage found in the NY40 genome was assumed to have been genetically modified, after site-specific integration. These, together with the data in the KHP30-like prophages of other H. pylori genomes, suggest that the lysogenic state of the KHP30-like phages is generally unstable.

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  • Characterization of Pseudomonas aeruginosa phage KPP21 belonging to family Podoviridae genus N4-like viruses isolated in Japan International journal

    Ryu Shigehisa, Jumpei Uchiyama, Shin ichiro Kato, Iyo Takemura-Uchiyama, Kotoe Yamaguchi, Reina Miyata, Takako Ujihara, Yoshihiko Sakaguchi, Noriaki Okamoto, Hidekatsu Shimakura, Masanori Daibata, Masahiro Sakaguchi, Shigenobu Matsuzaki

    Microbiology and Immunology   60 ( 1 )   64 - 67   2016.1

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    © 2016 The Societies and Wiley Publishing Asia Pty Ltd. Bacteriophages (phages) belonging to the family Podoviridae genus N4-like viruses have been used as therapeutic agent in phage therapy against Pseudomonas aeruginosa infections. P. aeruginosa phage KPP21 was isolated in Japan, and phylogenetically investigated the phages belonging to this viral genus. Morphological and genetic analyses confirmed that phage KPP21 belongs to the family Podoviridae genus N4-like viruses. Moreover, phylogenetic analyses based on putative DNA polymerase and major virion protein showed that P. aeruginosa phages belonging to the genus N4-like viruses are separated into two lineages and that phage KPP21 is in the same clade as phage LUZ7.

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  • Characterization of a novel Pseudomonas aeruginosa bacteriophage, KPP25, of the family Podoviridae Reviewed International journal

    Reina Miyata, Kotoe Yamaguchi, Jumpei Uchiyama, Ryu Shigehisa, Iyo Takemura-Uchiyama, Shin ichiro Kato, Takako Ujihara, Yoshihiko Sakaguchi, Masanori Daibata, Shigenobu Matsuzaki

    Virus Research   189   43 - 46   2014.8

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    Pseudomonas aeruginosa phages belonging to the family Podoviridae are one of the well-characterized phage groups. In this study, a novel P. aeruginosa phage, KPP25, was isolated and characterized. Phage KPP25's morphology was indicative of the family Podoviridae; however, analyses of the whole genome and the virion proteins suggested that it did not belong to any of the known podophage genera. Based on these analyses, phage KPP25 appears to be a novel podophage infecting P. aeruginosa. © 2014 Elsevier B.V.

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  • In silico analysis of AHJD-like viruses, Staphylococcus aureus phages S24-1 and S13 ', and study of phage S24-1 adsorption Reviewed International journal

    Jumpei Uchiyama, Iyo Takemura-Uchiyama, Shin-ichiro Kato, Miho Sato, Takako Ujihara, Hidehito Matsui, Hideaki Hanaki, Masanori Daibata, Shigenobu Matsuzaki

    MICROBIOLOGYOPEN   3 ( 2 )   257 - 270   2014.4

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    Staphylococcus aureus is a clinically important bacterium that is commensal in both humans and animals. Bacteriophage (phage) attachment to the host bacterial surface is an important process during phage infection, which involves interactions between phage receptor-binding proteins and host receptor molecules. However, little information is available on the receptor-binding protein of S. aureus phages. S. aureus virulent phages S24-1 and S13' (family Podoviridae, genus AHJD-like viruses) were isolated from sewage. In the present study, we investigated the receptor-binding protein of AHJD-like viruses using phage S24-1. First, based on a comparative genomic analysis of phages S24-1 and S13', open reading frame 16 (ORF16) of phage S24-1 was speculated to be the receptor-binding protein, which possibly determines the host range. Second, we demonstrated that this was the receptor-binding protein of phage S24-1. Third, our study suggested that wall teichoic acids in the cell walls of S. aureus are the main receptor molecules for ORF16 and phage S24-1. Finally, the C-terminal region of ORF16 may be essential for binding to S. aureus. These results strongly suggest that ORF16 of phage S24-1 and its homologs may be the receptor-binding proteins of AHJD-like viruses.

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  • Phage Therapy: Experiments Using Animal Infection Models Reviewed

    Matsuzaki, Shigenobu, Uchiyama, Jumpei, Takemura-Uchiyama, Iyo, Daibata, Masanori

    Phage Therapy   237 - 237   2014

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  • Draft genome sequence of Clostridium botulinum type B strain Osaka05, isolated from an infant patient with botulism in Japan Reviewed International journal

    Yoshihiko Sakaguchi, Koji Hosomi, Jumpei Uchiyama, Yoshitoshi Ogura, Kaoru Umeda, Masakiyo Sakaguchi, Tomoko Kohda, Masafumi Mukamoto, Naoaki Misawa, Shigenobu Matsuzaki, Tetsuya Hayashi, Shunji Kozaki

    Genome Announcements   2 ( 1 )   2014

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    © 2014 Sakaguchi et al. Clostridium botulinum strain Osaka05, which has been isolated from an infant patient with botulism in Japan, is the first strain producing botulinum neurotoxin subtype B6. Here, we report the draft genome sequence of C. botulinum Osaka05.

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  • Genome analysis of Pseudomonas aeruginosa bacteriophage KPP23, belonging to the family Siphoviridae Reviewed International journal

    Kotoe Yamaguchi, Reina Miyata, Ryu Shigehisa, Jumpei Uchiyama, Iyo Takemura-Uchiyama, Shin Ichiro Kato, Takako Ujihara, Yoshihiko Sakaguchi, Masanori Daibata, Shigenobu Matsuzaki

    Genome Announcements   2 ( 3 )   e00233 - 14   2014

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    © 2014 Yamaguchi et al. Bacteriophage (phage) therapy is expected to become an alternative therapy for Pseudomonas aeruginosa infections. P. aeruginosa phage KPP23 is a newly isolated phage belonging to the family Siphoviridae and may be a therapeutic phage candidate. We report its complete genome, which comprises 62,774 bp of double-stranded DNA containing 95 open reading frames.

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  • Evaluating efficacy of bacteriophage therapy against Staphylococcus aureus infections using a silkworm larval infection model International journal

    Iyo Takemura-Uchiyama, Jumpei Uchiyama, Shin ichiro Kato, Tetsuyoshi Inoue, Takako Ujihara, Naoya Ohara, Masanori Daibata, Shigenobu Matsuzaki

    FEMS Microbiology Letters   347 ( 1 )   52 - 60   2013.10

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    Silkworm larva has recently been recognized as an alternative model animal for higher mammals to evaluate the effects of antibiotics. In this study, we examined the efficacy of the bacteriophage (phage) therapy, which harnesses phages as antibacterial agents, against Staphylococcus aureus infections, using the silkworm larval infection model. Two newly isolated staphylococcal phages, S25-3 and S13′, were used as therapeutic phage candidates. They were assigned to two different lytic phage genera, Twort-like and AHJD-like viruses, based on their morphologies and the N-terminal amino acid sequences of the major capsid proteins. Both had a broad host range and strong lytic activity and showed preservative quality. Administration of these phages alone caused no adverse effects in the silkworm larvae. Moreover, the viruses showed life-prolonging effects in the silkworm larval infection model 10 min, 6 h, 12 h, and 24 h following infection. Such phage effects in the silkworm larval model were almost paralleled to the therapeutic efficacies in mouse models. These results suggest that phages S25-3 and S13′ are eligible as therapeutic candidates and that the silkworm larval model is valid for the evaluation of phage therapy as well as mouse models. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

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  • Synergistic bacteriolysis by bacteriophage φeF24C endolysin ORF9 and lantibiotic nisin and its application to pulsed-field gel electrophoretic analysis of Enterococcus faecalis Reviewed

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

    Annals of Microbiology   63 ( 3 )   1209 - 1211   2013.9

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    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.

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  • Pseudomonas aeruginosa Keratitis in Mice: Effects of Topical Bacteriophage KPP12 Administration Reviewed

    Ken Fukuda, Waka Ishida, Jumpei Uchiyama, Mohammad Rashel, Shin ichiro Kato, Tamae Morita, Asako Muraoka, Tamaki Sumi, Shigenobu Matsuzaki, Masanori Daibata, Atsuki Fukushima

    PLoS ONE   7 ( 10 )   e47742   2012.10

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    The therapeutic effects of bacteriophage (phage) KPP12 in Pseudomonas aeruginosa keratitis were investigated in mice. Morphological analysis showed that phage KPP12 is a member of the family Myoviridae, morphotype A1, and DNA sequence analysis revealed that phage KPP12 is similar to PB1-like viruses. Analysis of the phage KPP12 genome did not identify any genes related to drug resistance, pathogenicity or lysogenicity, and so phage KPP12 may be a good candidate for therapeutic. KPP12 showed a broad host range for P. aeruginosa strains isolated from clinical ophthalmic infections. Inoculation of the scarified cornea with P. aeruginosa caused severe keratitis and eventual corneal perforation. Subsequent single-dose administration of KPP12 eye-drops significantly improved disease outcome, and preserved the structural integrity and transparency of the infected cornea. KPP12 treatment resulted in the suppression of neutrophil infiltration and greatly enhanced bacterial clearance in the infected cornea. These results indicate that bacteriophage eye-drops may be a novel adjunctive or alternative therapeutic agent for the treatment of infectious keratitis secondary to antibiotic-resistant bacteria. © 2012 Fukuda et al.

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  • Complete Genome Sequences of Two Helicobacter pylori Bacteriophages Isolated from Japanese Patients Reviewed

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

    JOURNAL OF VIROLOGY   86 ( 20 )   11400 - 11401   2012.10

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    Helicobacter pylori causes peptic ulcers and gastric cancer, which lead to significantly higher morbidity in Japan than elsewhere in the world. As bacteriophage (phage) and host bacteria coevolve, the study of H. pylori phages is important to extend understanding of the evolution and pathogenesis of H. pylori. Here we report two complete genome sequences of H. pylori phages KHP30 and KHP40, which were released spontaneously from the most pathogenic East Asian-type isolates from Japanese patients.

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

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

    ARCHIVES OF VIROLOGY   157 ( 8 )   1587 - 1592   2012.8

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    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.

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  • Therapeutic Effects of Topical Bacteriophage KPP12 Administration on Pseudomonas aeruginosa Keratitis in Mice Reviewed

    Fukuda, Ken, Ishida, Waka, Uchiyama, Jumpei, Morita, Tamae, Harada, Yosuke, Sumi, Tamaki, Matsuzaki, Shigenobu, Daibata, Masanori, Fukushima, Atsuki

    Investigative Ophthalmology & Visual Science   53 ( 14 )   6201 - 6201   2012

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

    Christian Moro, Jumpei Uchiyama, Russ Chess-Williams

    Urology   78 ( 6 )   1442.e9 - 1442.e15   2011.12

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    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, Nω-nitro-l-arginine, or indomethacin. Carbachol (1 μM) 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 μM). Stretching of the tissue also caused an increase in the spontaneous contractile rate, and these responses were abolished by atropine (1 μM) and low concentrations of 4-diphenylacetoxy-N-methylpiperidine methiodide (10 nM). Darifenacin, oxybutynin, tolterodine, and solifenacin (1 μM) all significantly depressed the frequency responses to carbachol (1 μM). 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. © 2011 Elsevier Inc.

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  • Improved adsorption of an enterococcus faecalis bacteriophage φEF24C with a spontaneous point mutation Reviewed

    Jumpei Uchiyama, Iyo Takemura, Miho Satoh, Shin ichiro Kato, Takako Ujihara, Kazue Akechi, Shigenobu Matsuzaki, Masanori Daibata

    PLoS ONE   6 ( 10 )   e26648   2011.10

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    Some bacterial strains of the multidrug-resistant Gram-positive bacteria Enterococcus faecalis can significantly reduce the efficacy of conventional antimicrobial chemotherapy. Thus, the introduction of bacteriophage (phage) therapy is expected, where a phage is used as a bioagent to destroy bacteria. E. faecalis phage ΦEF24C is known to be a good candidate for a therapeutic phage against E. faecalis. However, this therapeutic phage still produces nonuniform antimicrobial effects with different bacterial strains of the same species and this might prove detrimental to its therapeutic effects. One solution to this problem is the preparation of mutant phages with higher activity, based on a scientific rationale. This study isolated and analyzed a spontaneous mutant phage, ΦEF24C-P2, which exhibited higher infectivity against various bacterial strains when compared with phage ΦEF24C. First, the improved bactericidal effects of phage ΦEF24C-P2 were attributable to its increased adsorption rate. Moreover, genomic sequence scanning revealed that phage ΦEF24C-P2 had a point mutation in orf31. Proteomic analysis showed that ORF31 (mw, 203 kDa) was present in structural components, and immunological analysis using rabbit-derived antibodies showed that it was a component of a long, flexible fine tail fiber extending from the tail end. Finally, phage ΦEF24C-P2 also showed higher bactericidal activity in human blood compared with phage ΦEF24C using the in vitro assay system. In conclusion, the therapeutic effects of phage ΦEF24C-P2 were improved by a point mutation in gene orf31, which encoded a tail fiber component. © 2011 Uchiyama et al.

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  • Presence of Merkel cell polyomavirus in Japanese cutaneous squamous cell carcinoma Reviewed

    Masanao Murakami, Masayuki Imajoh, Takuya Ikawa, Hideki Nakajima, Mikio Kamioka, Yuiko Nemoto, Takako Ujihara, Jumpei Uchiyama, Shigenobu Matsuzaki, Shigetoshi Sano, Masanori Daibata

    Journal of Clinical Virology   50 ( 1 )   37 - 41   2011.1

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    Background: Merkel cell polyomavirus (MCPyV) was first identified in Merkel cell carcinoma (MCC) as a new tumor virus. Studies have also reported differing frequencies of MCPyV detection in other skin cancers in western countries. Objectives: Little is known about geographical differences of MCPyV prevalence in non-MCC tumors. We examined the existence of MCPyV in non-MCC skin cancers including squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) in Japanese patients. Study design: Paraffin-embedded tissues of cutaneous SCC (n= 30) and BCC (n= 10) from Japanese patients were tested for the presence of MCPyV by polymerase chain reaction (PCR) with primer sets directed against the genes encoding large-T antigen 3 (LT3) and viral protein 1 (VP1). This was followed by DNA fragment sequencing and immunohistochemistry. Results: PCR analysis targeting the LT3 gene showed that the viral sequences were found in 4 of 30 (13%) SCC cases. Nested PCR detected the VP1 region in four cases. Sequencing analysis of these PCR-amplified fragments showed a close homology to the previously published MCPyV sequences. Immunohistochemistry with the monoclonal antibody to MCPyV LT-antigen showed positive staining in 2 of 4 LT3 PCR-positive cases. On the other hand, our BCC samples were all negative for MCPyV. Conclusion: This study suggested that Japanese cutaneous SCC is infrequently associated with MCPyV. Further worldwide epidemiological surveys are warranted to determine the possible association of MCPyV with pathogenesis of non-MCC skin cancers. © 2010 Elsevier B.V.

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  • Characterization of lytic enzyme open reading frame 9 (ORF9) derived from Enterococcus faecalis bacteriophage $φ$EF24C Reviewed

    Uchiyama, Jumpei, Takemura, Iyo, Hayashi, Ikue, Matsuzaki, Shigenobu, Satoh, Miho, Ujihara, Takako, Murakami, Masanao, Imajoh, Masayuki, Sugai, Motoyuki, Daibata, Masanori

    Applied and environmental microbiology   77 ( 2 )   580 - 585   2011

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    In bacteriophage (phage) therapy against Gram-positive bacteria, such as Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis, members of a genus of SPO1-like viruses are typically employed because of their extreme virulence and broad host spectrum. Phage φEF24C, which is a SPO1-like virus infecting E. faecalis, has previously been characterized as a therapeutic phage candidate. In addition to the phage itself, phage endolysin is also recognized as an effective antimicrobial agent. In this study, a putative endolysin gene (orf9) of E. faecalis phage φEF24C was analyzed in silico, and its activity was characterized using the recombinant form. First, bioinformatics analysis predicted that the open reading frame 9 (ORF9) protein is N-acetylmuramoyl-L-alanine amidase. Second, bacteriolytic and bactericidal activities of ORF9 against E. faecalis were confirmed by zymography, decrease of peptidoglycan turbidity, decrease of the viable count, and morphological analysis of ORF9-treated cells. Third, ORF9 did not appear to require Zn 2+ ions for its activity, contrary to the bioinformatics prediction of a Zn2+ ion requirement. Fourth, the lytic spectrum was from 97.1% (34 out of 35 strains, including vancomycin-resistant strains) of E. faecalis strains to 60% (6 out of 10 strains) of Enterococcus faecium strains. Fifth, N-acetylmuramoyl-L-alanine amidase activity of ORF9 was confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and the subsequent MALDI-postsource decay (PSD) analyses. Finally, functional analysis using N- or C-terminally deleted ORF9 mutants suggested that a complete ORF9 molecule is essential for its activity. These results suggested that ORF9 is an endolysin of phage φEF24C and can be a therapeutic alternative to antibiotics. Copyright © 2011, American Society for Microbiology.

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  • Isolation and characterization of a novel Staphylococcus aureus bacteriophage, φMR25, and its therapeutic potential Reviewed

    Hiroshi Hoshiba, Jumpei Uchiyama, Shin ichiro Kato, Takako Ujihara, Asako Muraoka, Masanori Daibata, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    Archives of Virology   155 ( 4 )   545 - 552   2010.4

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    A novel bacteriophage, φMR25, was isolated from a lysogenic Staphylococcus aureus strain by mitomycin C induction. Its biological features were analyzed in comparison with φMR11, which was described previously as a prototype therapeutic phage. φMR25 is morphologically similar to φMR11 (morphotype B1 of family Myoviridae) but has a broader host range than φMR11 on S. aureus strains. φMR25 can also multiply on S. aureus lysogens of φMR11. Its DNA is 44,342 bp in size, is predicted to include 70 open reading frames, and does not contain genes related to toxin or drug resistance. The lysogenic module and most of the putative virion protein genes are completely different from those of φMR11. In spite of their genetic diversity, intraperitoneal administration of φMR25 rescued mice inoculated with a lethal dose of S. aureus, as was the case for φMR11. These results suggest that φMR25 could be another candidate phage to treat S. aureus infection. © Springer-Verlag 2010.

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  • Morphological and genetic analysis of three bacteriophages of Serratia marcescens isolated from environmental water: Research Letter Reviewed

    Kenshi Matsushita, Jumpei Uchiyama, Shin Ichiro Kato, Takako Ujihara, Hiroshi Hoshiba, Shigeyoshi Sugihara, Asako Muraoka, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    FEMS Microbiology Letters   291 ( 2 )   201 - 208   2009.2

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    Increases in multidrug-resistant strains of Serratia marcescens are of great concern in pediatrics, especially in neonatal intensive care units. In the search for bacteriophages to control infectious diseases caused by multidrug-resistant S. marcescens, three phages (KSP20, KSP90, and KSP100) were isolated from environmental water and were characterized morphologically and genetically. KSP20 and KSP90 belonged to morphotype A1 of the family Myoviridae, and KSP100 belonged to morphotype C3 of the family Podoviridae. Analysis of the DNA region coding virion proteins, together with their morphological features, indicated that KSP20, KSP90, and KSP100 were related to the P2-like phage (temperate), T4-type phage (virulent), and phiEco32 phage (virulent), respectively. Based on amino acid sequences of the major capsid protein, KSP90 formed a new branch with a Stenotrophomonas maltophilia phage, Smp14, in the T4-type phage phylogeny. Both Smp14 and phiEco32 have been reported as potential therapeutic phages. These results suggest that KSP90 and KSP100 may be candidate therapeutic phages to control S. marcescens infection. © 2008 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

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  • Characteristics of a novel Pseudomonas aeruginosa bacteriophage, PAJU2, which is genetically related to bacteriophage D3 Reviewed

    Jumpei Uchiyama, Mohammad Rashel, Tetsuya Matsumoto, Yoshinobu Sumiyama, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    Virus Research   139 ( 1 )   131 - 134   2009.1

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    Pseudomonas aeruginosa bacteriophage (phage) is one of the most taxonomically and genetically diverse phages. Although phage D3 is one of well-studied P. aeruginosa phages, no D3-related P. aeruginosa phage has been reported. We report a novel P. aeruginosa siphovirus, PAJU2, which is genetically related to but morphology distinct (highly elongated head) from phage D3. A PAJU2 capsid protein, Orf3, is thought to be synthesized as a protein fused to a prohead protease and is autocatalytically cleaved, which may form the head chain mail. Despite such morphological differences, PAJU2 is expected to be a useful genetic reference for phage D3. © 2008 Elsevier B.V. All rights reserved.

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  • Tail-associated structural protein gp61 of Staphylococcus aureus phage φmR11 has bifunctional lytic activity Reviewed

    Mohammad Rashel, Jumpei Uchiyama, Iyo Takemura, Hiroshi Hoshiba, Takako Ujihara, Hiroyoshi Takatsuji, Koichi Honke, Shigenobu Matsuzaki

    FEMS Microbiology Letters   284 ( 1 )   9 - 16   2008.7

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    A tailed bacteriophage, φMR11 (siphovirus), was selected as a candidate therapeutic phage against Staphylococcus aureus infections. Gene 61, one of the 67 ORFs identified, is located in the morphogenic module. The gene product (gp61) has lytic domains homologous to CHAP (corresponding to an amidase function) at its N-terminus and lysozyme subfamily 2 (LYZ2) at its C-terminus. Each domain of gp61 was purified as a recombinant protein. Both the amidase [amino acids (aa) 1-150] and the lysozyme (aa 401-624) domains but not the linker domain (aa 151-400) caused efficient lysis of S. aureus. Immunoelectron microscopy localized gp61 to the tail tip of the φMR11 phage. These data strongly suggest that gp61 is a tail-associated lytic factor involved in local cell-wall degradation, allowing the subsequent injection of φMR11 DNA into the host cytoplasm. Staphylococcus aureus lysogenized with φMR11 was also lysed by both proteins. Staphylococcus aureus strains on which φMR11 phage can only produce spots but not plaques were also lysed by each protein, indicating that gp61 may be involved in 'lysis from without'. This is the first report of the presence of a tail-associated virion protein that acts as a lysin, in an S. aureus phage. © 2008 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

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  • A novel site-specific recombination system derived from bacteriophage φ{symbol}MR11 Reviewed

    Mohammad Rashel, Jumpei Uchiyama, Takako Ujihara, Iyo Takemura, Hiroshi Hoshiba, Shigenobu Matsuzaki

    Biochemical and Biophysical Research Communications   368 ( 2 )   192 - 198   2008.4

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    We report identification of a novel site-specific DNA recombination system that functions in both in vivo and in vitro, derived from lysogenic Staphylococcus aureus phage φ{symbol}MR11. In silico analysis of the φ{symbol}MR11 genome indicated orf1 as a putative integrase gene. Phage and bacterial attachment sites (attP and attB, respectively) and attachment junctions were determined and their nucleotide sequences decoded. Sequences of attP and attB were mostly different to each other except for a two bp common core that was the crossover point. We found several inverted repeats adjacent to the core sequence of attP as potential protein binding sites. The precise and efficient integration properties of φ{symbol}MR11 integrase were shown on attP and attB in Escherichia coli and the minimum size of attP was found to be 34 bp. In in vitro assays using crude or purified integrase, only buffer and substrate DNAs were required for the recombination reaction, indicating that other bacterially encoded factors are not essential for activity. © 2008 Elsevier Inc. All rights reserved.

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  • T-even-related bacteriophages as candidates for treatment of Escherichia coli urinary tract infections Reviewed

    H. Nishikawa, M. Yasuda, J. Uchiyama, M. Rashel, Y. Maeda, I. Takemura, S. Sugihara, T. Ujihara, Y. Shimizu, T. Shuin, S. Matsuzaki

    Archives of Virology   153 ( 3 )   507 - 515   2008.3

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    Multidrug-resistant uropathogenic Escherichia coli (UPEC) is increasing gradually on a worldwide scale. We therefore examined the possibility of bacteriophage (phage) therapy for urinary tract infections (UTIs) caused by the UPEC strains as an alternative to chemotherapy. In addition to the well-known T4 phage, KEP10, which was newly isolated, was used as a therapeutic phage candidate. KEP10 showed a broader bacteriolytic spectrum (67%) for UPEC strains than T4 (14%). Morphological and genetic analyses showed that KEP10 resembles phage T4. Phages T4 and KEP10 injected into the peritoneal cavity of mice were distributed immediately to all organs examined and maintained a high titer for at least 24 h. They were stable in the urine of both mice and humans for 24 h at 37°C. Administration of these phages into the peritoneal cavity caused a marked decrease in the mortality of mice inoculated transurethrally with a UPEC strain, whereas most of the control mice died within a few days of bacterial infection. Inoculation with phage alone produced no adverse effects attributable to the phage per se. The present study experimentally demonstrated the therapeutic potential of phage for E. coli-induced UTIs, and T-even-related phages may be suitable candidates with which to treat them. © 2008 Springer-Verlag.

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  • Efficacy of bacteriophage therapy against gut-derived sepsis caused by Pseudomonas aeruginosa in mice Reviewed

    Ryohei Watanabe, Tetsuya Matsumoto, Go Sano, Yoshikazu Ishii, Kazuhiro Tateda, Yoshinobu Sumiyama, Jumpei Uchiyama, Shingo Sakurai, Shigenobu Matsuzaki, Shosuke Imai, Keizo Yamaguchi

    ANTIMICROBIAL AGENTS AND CHEMOTHERAPY   51 ( 2 )   446 - 452   2007.2

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    We evaluated the efficacy of bacteriophage (phage) therapy by using a murine model of gut-derived sepsis caused by Pseudomonas aeruginosa that closely resembles the clinical pathophysiology of septicemia in humans. Oral administration of a newly isolated lytic phage strain (KPP10) significantly protected mice against mortality (survival rates, 66.7% for the phage-treated group versus 0% for the saline-treated control group; P < 0.01). Mice treated with phage also had lower numbers of viable P. aeruginosa cells in their blood, liver, and spleen. The levels of inflammatory cytokines (tumor necrosis factor alpha TNF-alpha, interleukin-1 beta [IL-1 beta], and IL-6) in blood and liver were significantly lower in phage-treated mice than in phage-untreated mice. The number of viable P. aeruginosa cells in fecal matter in the gastrointestinal tract was significantly lower in phage-treated mice than in the saline-treated control mice. We also studied the efficacy of phage treatment for intraperitoneal infection caused by P. aeruginosa and found that phage treatment significantly improved the survival of mice, but only under limited experimental conditions. In conclusion, our findings suggest that oral administration of phage may be effective against gut-derived sepsis caused by P. aeruginosa.

    DOI: 10.1128/AAC.00635-06

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  • Efficient elimination of multidrug-resistant Staphylococcus aureus by cloned lysin derived from bacteriophage φMR11 Reviewed

    Mohammad Rashel, Jumpei Uchiyama, Takako Ujihara, Yoshio Uehara, Shu Kuramoto, Shigeyoshi Sugihara, Ken Ichi Yagyu, Asako Muraoka, Motoyuki Sugai, Keiichi Hiramatsu, Koichi Honke, Shigenobu Matsuzaki

    Journal of Infectious Diseases   196 ( 8 )   1237 - 1247   2007

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    We report the successful purification of a cloned lysin encoded by the novel Staphylococcus aureus bacteriophage φMR11. The lysin, designated MV-L, rapidly and completely lysed cells of a number of S. aureus strains tested, including methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus and a subset of vancomycin-intermediate S. aureus (VISA) in growing conditions. MV-L-mediated killing is specific to S. aureus and not to other species, except for S. simulans. MV-L exerted its staphylocidal effect synergistically with glycopeptide antibiotics against VISA. MV-L efficiently eliminated MRSA that had been artificially inoculated into the nares of mice. The intraperitoneal administration of MV-L also protected mice against MRSA septic death, without any harmful effects. Although MV-L evoked detectable levels of a humoral response in mice, the antibodies did not abolish the bacteriolytic activity. These results indicate that MV-L might be useful as a powerful therapeutic agent against multidrug-resistant S. aureus infections. © 2007 by the Infectious Diseases Society of America. All rights reserved.

    DOI: 10.1086/521305

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  • ファージを使った医療(第4回)ファージと生体 Reviewed

    内山 淳平

    Veterinary immunology for practitioners : 獣医免疫アレルギー学専門誌   6 ( 1 )   18 - 24   2021.1

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    Other Link: http://search.jamas.or.jp/link/ui/2021130325

  • R&D of phage therapy to MRSA infections: (I) Analysis of phage culture by design of experiment Invited

    内山淳平

    日本細菌学雑誌(Web)   76 ( 1 )   2021

  • ファージを使った医療(第3回) ファージを利用した細菌検査法 Invited Reviewed

    内山 淳平

    Veterinary Immunology for Practitioners   5 ( 4 )   30 - 37   2020.10

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  • ファージを使った医療(第2回)ファージ療法 Invited Reviewed

    内山 淳平

    Veterinary immunology for practitioners : 獣医免疫アレルギー学専門誌   5 ( 3 )   36 - 41   2020.7

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    Other Link: http://search.jamas.or.jp/link/ui/2020311001

  • ファージを使った医療(第1回)ファージとは何か? Invited Reviewed

    内山 淳平

    Veterinary immunology for practitioners : 獣医免疫アレルギー学専門誌   5 ( 2 )   36 - 42   2020.4

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    Other Link: http://search.jamas.or.jp/link/ui/2020189278

  • マイクロバイオーム解析の基礎:細菌叢解析を中心に Invited

    内山淳平

    ペット栄養学会誌   21 ( 2 )   S11 - S11   2018

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    DOI: 10.11266/jpan.21.2_S11

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  • 健常犬の糞便中から分離した乳酸菌AZABU株はヒト単球細胞をM2bマクロファージに分化誘導する

    安田 伊武希, 金木 真央, 竹田 志郎, 内山 淳平, 福山 朋季

    日本獣医学会学術集会講演要旨集   165回   [J2A - 02]   2022.9

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  • イヌにおける胆嚢の常在細菌叢の存在の検討と肝疾患に関連した細菌叢解析

    経田 真梨菜, 根尾 櫻子, 内山 伊代, 内山 淳平, 村上 裕信, 島 綾香, 茅沼 秀樹, 横山 大希, 高木 哲, 金井 詠一, 久末 正晴, 葛原 早瑛

    日本獣医学会学術集会講演要旨集   165回   [HS1P - 15]   2022.9

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  • 分娩後子宮捻転を発症した乳牛の子宮内細菌叢解析

    風間 啓, 内山 淳平, 恩田 賢

    日本獣医学会学術集会講演要旨集   165回   [HL1P - 01]   2022.9

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  • Potential therapeutic effects of ozone water on pyoderma and atopic dermatitis in dogs

    Kaneki Mao, Takahashi Miyu, Uchiyama Jumpei, Ano Tetsuya, Fukuyama Tomoki

    Proceedings for Annual Meeting of The Japanese Pharmacological Society   95   1-SS-64   2022

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    Ozone water is currently utilized for antibacterial and antiviral purposes under the pandemic of COVID-19 due to its strong oxidation effect without safety problems. Therefore, clinical applications of ozone water for cutaneous diseases such as atopic dermatitis (AD) and pyoderma are expected recently. The aim of this project is to apply the ozone water for AD and pyoderma in dogs. We here examined the antibacterial effects of ozone water using staphylococci in in vitro setting, and anti-allergic effects of ozone water in AD mouse model.

    We first examined the bactericidal effect of ozone water on resident staphylococci on skin. Staphylococcus aureus, which is an exacerbating factor in human AD, and S. lentus collected from AD mouse model was treated with ozone water and the number of bacteria was determined. Our findings indicated that ozone water showed a significant bactericidal effect against both staphylococcal species . We are currently investigating the efficacy of ozone water against staphylococci isolated from AD and pyoderma dogs. In vivo experiment with AD mouse model is also underway to look for the therapeutic effects of ozone water with impact on inflammatory and itch responses.

    DOI: 10.1254/jpssuppl.95.0_1-ss-64

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  • Ozone water as a potential preventive and therapeutic option for periodontal disease in dogs

    Tomoki Fukuyama, Kaneki Mao, Oohira Chiharu, Uchiyama Jumpei, Ano Tetsuya

    Proceedings for Annual Meeting of The Japanese Pharmacological Society   95   2-O-090   2022

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    Periodontal disease, which is majorly caused by Porphyromonas gulae, is extremely common in dogs, and is associated with serious systemic diseases such as kidney failure, heart disease and immune disorder. Ozon water is ozone-gas-dissolved water, which has antimicrobial activity by oxidizing molecules. However, both therapeutic and preventive effects of ozone water for periodontal disease in dogs are still unclear. We herein examined the antibacterial and anti-inflammatory effects of ozone water in vitro using P. gulae. Our results indicated that treatment of ozone water significantly inhibited the growth of P. gulae until 24h post ozone water treatment as compared to purified water treatment. Ozone water also showed the anti-inflammatory effects in P. gulae-infected human gingival epithelial cells (Ca9-22). Secretions of IL-1β and TNFα were significantly suppressed by ozone water treatment compared to the treatment of purified water. As IL-1β and TNFα activate the osteoclast and finally promote bone resorption in the oral cavity, our findings imply the potential therapeutic and preventive effects of ozone water in canine periodontal disease. We are currently investigating the in vivo preventive and therapeutic effects of ozone water in dogs.

    DOI: 10.1254/jpssuppl.95.0_2-o-090

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  • 正常犬の糞便中から分離した乳酸菌AZABU株生菌の経口投与がアトピー性皮膚炎モデルマウスの炎症およびそう痒に及ぼす効果検討

    早川 千春, 内山 淳平, 久末 正晴, 根尾 櫻子, 村上 裕信, 五十嵐 寛高, 宮武 昌一郎, 阪口 雅弘, 福山 朋季

    日本獣医学会学術集会講演要旨集   164回   [JO - 17]   2021.9

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  • 妊娠後期のB群連鎖球菌検査の精度を改善のためのファージ酵素の利用

    内山 淳平, 松井 秀仁, 小方 雅也, 那須川 忠弥, 内山 伊代, 加藤 伸一郎, 今西 市朗, 東出 正人, 花木 秀明

    日本獣医学会学術集会講演要旨集   164回   [DBO - 24]   2021.9

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  • アレルゲン不活化剤の新規評価法としてのドットブロット法の開発

    吉田 愛美, 水上 圭二郎, 蔵田 圭吾, 那須川 忠弥, 内山 淳平, 阪口 雅弘

    アレルギー   70 ( 6-7 )   875 - 875   2021.8

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  • アレルゲン不活化剤の新規評価法としてのドットブロット法の開発

    吉田 愛美, 水上 圭二郎, 蔵田 圭吾, 那須川 忠弥, 内山 淳平, 阪口 雅弘

    アレルギー   70 ( 6-7 )   875 - 875   2021.8

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  • 膿痂疹の自然発症動物モデル(イヌ表在性膿皮症)に対するファージ由来酵素エンドライシンの検討

    今西 市朗, 西藤 公司, 朝比奈 良太, 林 俊治, 津久井 利広, 内山 淳平

    感染症学雑誌   95 ( 臨増 )   313 - 313   2021.4

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  • GBS検査の精度を改善するためのファージ酵素エンドライシンの利用

    内山 淳平, 松井 秀仁, 小方 雅也, 那須川 忠弥, 内山 伊代, 東出 正人, 阪口 雅弘, 花木 秀明

    感染症学雑誌   95 ( 臨増 )   313 - 313   2021.4

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  • GBS検査の精度を改善するためのファージ酵素エンドライシンの利用

    内山 淳平, 松井 秀仁, 小方 雅也, 那須川 忠弥, 内山 伊代, 東出 正人, 阪口 雅弘, 花木 秀明

    感染症学雑誌   95 ( 臨増 )   313 - 313   2021.4

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  • 膿痂疹の自然発症動物モデル(イヌ表在性膿皮症)に対するファージ由来酵素エンドライシンの検討

    今西 市朗, 西藤 公司, 朝比奈 良太, 林 俊治, 津久井 利広, 内山 淳平

    感染症学雑誌   95 ( 臨増 )   313 - 313   2021.4

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  • 密度勾配超遠心法とフローサイトメトリーを併用したグラム陽性細菌が産生する膜小胞試料の調製法の検討

    那須川 忠弥, 杉本 良輔, 内山 淳平, 内山 伊代, 村上 裕信, 福田 憲, 松崎 茂展, 阪口 雅弘

    日本細菌学雑誌   76 ( 1 )   83 - 83   2021.2

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  • 黄色ブドウ球菌に対するファージ療法の研究開発 実験計画法によるファージ培養の解析 Invited

    内山 淳平

    日本細菌学雑誌   76 ( 1 )   9 - 9   2021.2

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  • 密度勾配超遠心法とフローサイトメトリーを併用したグラム陽性細菌が産生する膜小胞試料の調製法の検討

    那須川 忠弥, 杉本 良輔, 内山 淳平, 内山 伊代, 村上 裕信, 福田 憲, 松崎 茂展, 阪口 雅弘

    日本細菌学雑誌   76 ( 1 )   83 - 83   2021.2

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  • ファージ由来溶菌酵素S25-3LYSのイヌ表在性膿皮症に対する臨床応用性の検討

    今西 市朗, 西藤 公司, 朝比奈 良太, 林 俊治, 津久井 利弘, 内山 淳平

    日本細菌学雑誌   76 ( 1 )   123 - 123   2021.2

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  • 黄色ブドウ球菌に対するファージ療法の研究開発 実験計画法によるファージ培養の解析

    内山 淳平

    日本細菌学雑誌   76 ( 1 )   9 - 9   2021.2

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  • GBS検査を改善するエンドライシンの新規利用法検討

    小方 雅也, 松井 秀仁, 那須川 忠弥, 内山 伊代, 東出 正人, 阪口 雅弘, 花木 秀明, 内山 淳平

    日本細菌学雑誌   76 ( 1 )   66 - 66   2021.2

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  • ファージ由来溶菌酵素S25-3LYSのイヌ表在性膿皮症に対する臨床応用性の検討

    今西 市朗, 西藤 公司, 朝比奈 良太, 林 俊治, 津久井 利弘, 内山 淳平

    日本細菌学雑誌   76 ( 1 )   123 - 123   2021.2

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  • 菌種同定に難渋した感染性心内膜炎の1例

    岡村 駿, 明城 正博, 杉下 靖之, 田部井 史子, 佐藤 優佳理, 今西 市朗, 内山 淳平, 伊從 慶太, 曾和 裕之

    日本内科学会関東地方会   666回   30 - 30   2021.2

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  • GBS検査を改善するエンドライシンの新規利用法検討

    小方 雅也, 松井 秀仁, 那須川 忠弥, 内山 伊代, 東出 正人, 阪口 雅弘, 花木 秀明, 内山 淳平

    日本細菌学雑誌   76 ( 1 )   66 - 66   2021.2

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  • GBS培養検査で問題となる偽陰性を改善する新規選択的抗菌剤の技術開発

    小方 雅也, 松井 秀仁, 那須川 忠弥, 内山 伊代, 東出 正人, 阪口 雅弘, 花木 秀明, 内山 淳平

    神奈川医学会雑誌   48 ( 1 )   69 - 69   2021.1

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  • Examination of novel application of endolysin to improve GBS test

    小方雅也, 松井秀仁, 那須川忠弥, 内山伊代, 東出正人, 阪口雅弘, 花木秀明, 内山淳平

    日本細菌学雑誌(Web)   76 ( 1 )   2021

  • 強制劣化したソーセージ細菌叢の解析とくん液添加による影響

    竹田志郎, 内山淳平, 木下由貴, 水野谷航, 坂田亮一

    日本食肉研究会総会提出議案及び大会講演要旨   62nd (CD-ROM)   2021

  • GBS検査の精度を改善するためのファージ酵素エンドライシンの利用

    内山淳平, 松井秀仁, 小方雅也, 那須川忠弥, 内山伊代, 東出正人, 阪口雅弘, 花木秀明

    日本化学療法学会雑誌   69 ( Supplement-A )   2021

  • Clinical applicability of phage-derived lytic enzyme S25-3LYS to canine superficial pyoderma

    今西市朗, 西藤公司, 朝比奈良太, 林俊治, 津久井利弘, 内山淳平

    日本細菌学雑誌(Web)   76 ( 1 )   2021

  • Purification of membrane vesicles from Gram-positive bacteria using flow cytometry

    那須川忠弥, 杉本良輔, 内山淳平, 内山伊代, 村上裕信, 福田憲, 松崎茂展, 阪口雅弘

    日本細菌学雑誌(Web)   76 ( 1 )   2021

  • Colorimetric detection of bacteria pathogens through aggregation of gold nanoparticles induced by thiolated bacteriophages

    山下智史, 仁子陽輔, 波多野慎悟, 渡辺茂, 内山伊代, 内山淳平, 松崎茂展

    日本化学会春季年会講演予稿集(Web)   101st   2021

  • 膿痂疹の自然発症動物モデル(イヌ表在性膿皮症)に対するファージ由来酵素エンドライシンの検討

    今西市朗, 西藤公司, 朝比奈良太, 林俊治, 津久井利広, 内山淳平

    日本化学療法学会雑誌   69 ( Supplement-A )   2021

  • 酸素ナノバブル水の抗腫瘍作用に関する基礎的研究

    永根大幹, 丹羽智瑛, 内山淳平, 稲波修, 山下匡

    日本酸化ストレス学会学術集会プログラム・抄録集   74th (CD-ROM)   2021

  • 新規腸球菌バクテリオファージによる腸球菌眼内炎に対するファージ療法の効果

    岸本達真, 石田わか, 鈴木崇, 内山淳平, 松崎茂展, 大畑雅典, 戸所大輔, 福田憲, 福島敦樹

    日本眼感染症学会・日本眼炎症学会・日本コンタクトレンズ学会総会・日本涙道・涙液学会プログラム・講演抄録集   57th-54th-63rd-9th   2021

  • 正常犬の糞便中から分離した乳酸菌AZABU株生菌の経口投与がアトピー性皮膚炎モデルマウスの炎症および掻痒に及ぼす効果検討

    早川千春, 内山淳平, 久末正晴, 根尾櫻子, 村上裕信, 五十嵐寛高, 宮武昌一郎, 阪口雅弘, 福山朋季

    日本獣医学会学術集会講演要旨集   164th (CD-ROM)   [JO - 17]   2021

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  • 妊娠後期のB群連鎖球菌検査の精度を改善のためのファージ酵素の利用

    内山淳平, 内山淳平, 松井秀仁, 小方雅也, 那須川忠弥, 内山伊代, 加藤伸一郎, 今西市朗, 東出正人, 花木秀明

    日本獣医学会学術集会講演要旨集   164th (CD-ROM)   [DBO - 24]   2021

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  • Intracellular signaling pathways which can regulate the replication of African pygmy hedgehog adenovirus

    文榕鐸, 落合秀治, 内山淳平, 大澤南菜子, 大場真己, 片山幸枝, 黎凱欣, 大松勉, 田向健一, 鈴木馨, 斑目広郎, 牧野伸治, 水谷哲也

    日本ウイルス学会学術集会プログラム・予稿集(Web)   68th   2021

  • Staphylococcus pseudintermediusに対する新規エンドライシンの分離と解析

    金木 真央, 内山 淳平, 松崎 茂展, 内山 伊代, 小方 雅也, 阪口 雅弘

    日本獣医学会学術集会講演要旨集   163回   215 - 215   2020.10

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  • 犬の糞便細菌叢移植における移植経路についての基礎的検討

    伊藤 哲之, 五十嵐 寛高, 内山 淳平, 新田 卓, 久末 正晴

    日本獣医学会学術集会講演要旨集   163回   285 - 285   2020.10

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  • イヌの純血種コロニーを利用したアトピー性皮膚炎に関連した口腔・腸内細菌叢の変動の解析

    海野 朝香, 今西 市朗, 水上 圭二郎, 大隅 尊史, 五十嵐 寛高, 村上 裕信, 島 綾香, 石原 玄基, 宇根 有美, 阪口 雅弘, 内山 淳平

    日本獣医学会学術集会講演要旨集   163回   216 - 216   2020.10

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  • Staphylococcus pseudintermediusのイヌアトピー性皮膚炎増悪因子の探索

    鶴井 大幹, 島倉 秀勝, 内山 淳平, 今西 市朗, 坂本 修士, 樋口 琢磨, 伊從 慶太, 下池 健太, 藤村 正人, 阪口 雅弘

    日本獣医学会学術集会講演要旨集   163回   216 - 216   2020.10

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  • フローサイトメーターを利用したグラム陽性細菌の膜小胞の新規分離法

    杉本 良輔, 那須川 忠弥, 内山 淳平, 内山 伊代, 村上 裕信, 福田 憲, 松崎 茂展, 阪口 雅弘

    日本獣医学会学術集会講演要旨集   163回   215 - 215   2020.10

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  • 建築様式および換気方式の異なる住宅における花粉侵入数の検討

    白井 秀治, 山口 裕礼, 小原 雄大, 吉田 誠, 渡邉 直人, 牧野 荘平, 内山 淳平, 阪口 雅弘

    アレルギー   69 ( 臨時増刊号 )   299 - 299   2020.10

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  • 感染細胞のシグナル伝達の活性化がハリネズミアデノウイルス増殖に及ぼす影響

    文 榕鐸, 落合 秀治, 内山 淳平, 大澤 南菜子, 田向 健一, 鈴木 馨, 大場 真己, 片山 幸枝, 斑目 広郎, 牧野 伸治, 水谷 哲也

    日本獣医学会学術集会講演要旨集   163回   235 - 235   2020.10

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  • 建築様式および換気方式の異なる住宅における花粉侵入数の検討

    白井 秀治, 山口 裕礼, 小原 雄大, 吉田 誠, 渡邉 直人, 牧野 荘平, 内山 淳平, 阪口 雅弘

    アレルギー   69 ( 臨時増刊号 )   299 - 299   2020.10

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  • エンドリシン製剤開発に向けて ブドウ球菌ファージS6の新規エンドリシンの分離

    金木 真央, 内山 淳平, 松崎 茂展, 内山 伊代, 小方 雅也, 林 俊治, 阪口 雅弘

    神奈川医学会雑誌   47 ( 2 )   233 - 234   2020.7

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  • GBS培養検査の精度を改善する選択的抗菌剤の技術開発

    小方 雅也, 松井 秀仁, 内山 淳平, 東出 正人, 内山 伊代, 金木 真央, 松崎 茂展, 花木 秀明, 阪口 雅弘

    神奈川医学会雑誌   47 ( 2 )   233 - 233   2020.7

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  • エンドリシン製剤開発に向けて ブドウ球菌ファージS6の新規エンドリシンの分離

    金木 真央, 内山 淳平, 松崎 茂展, 内山 伊代, 小方 雅也, 林 俊治, 阪口 雅弘

    神奈川医学会雑誌   47 ( 2 )   233 - 234   2020.7

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  • GBS培養検査の精度を改善する選択的抗菌剤の技術開発

    小方 雅也, 松井 秀仁, 内山 淳平, 東出 正人, 内山 伊代, 金木 真央, 松崎 茂展, 花木 秀明, 阪口 雅弘

    神奈川医学会雑誌   47 ( 2 )   233 - 233   2020.7

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  • Omics analysis in fecal transplantation therapy for Clostridioides difficile infection

    阪口義彦, 後藤和義, 妹尾充敏, 武晃, 内山淳平, 尾崎隼人, 城代康貴, 林俊治, 大宮直木, 加藤はる

    日本薬学会年会要旨集(CD-ROM)   140年会   28P - am101   2020.3

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  • Omics analysis in fecal transplantation therapy for Clostridioides difficile infection

    阪口義彦, 後藤和義, 妹尾充敏, 武晃, 内山淳平, 尾崎隼人, 城代康貴, 林俊治, 大宮直木, 加藤はる

    日本薬学会年会要旨集(CD-ROM)   140年会   28P - am101   2020.3

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  • ファージ・シンポジウム-忘れられていた治療法の復活- 薬剤耐性菌に対するファージセラピーの概説 Invited

    内山 淳平

    感染症学雑誌   94 ( 臨増 )   229 - 229   2020.3

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  • ファージ・シンポジウム-忘れられていた治療法の復活- 薬剤耐性菌に対するファージセラピーの概説 Invited

    内山 淳平

    感染症学雑誌   94 ( 臨増 )   229 - 229   2020.3

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  • 妊婦のGBSスクリーニングテストの検査精度を向上する試薬の開発

    小方 雅也, 内山 淳平, 松井 秀仁, 内山 伊代, 那須川 忠弥, 松崎 茂展, 花木 秀明, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   68 - 68   2020.1

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  • 妊婦のGBSスクリーニングテストの検査精度を向上する試薬の開発

    小方 雅也, 内山 淳平, 松井 秀仁, 内山 伊代, 那須川 忠弥, 松崎 茂展, 花木 秀明, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   68 - 68   2020.1

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  • グラム陽性細菌が産生する膜小胞の精製法の検討

    那須川 忠弥, 杉本 良輔, 島倉 秀勝, 小方 雅也, 福田 憲, 松崎 茂展, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   73 - 73   2020.1

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  • 牛白血病ウイルス感染による乳牛の腸内細菌叢の解析

    内山 淳平, 村上 裕信, 佐藤 礼一郎, 水上 圭二郎, 鈴木 武人, 島 綾香, 石原 玄基, 阪口 義彦, 曽川 一幸, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   126 - 126   2020.1

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  • イヌにおけるアトピー性皮膚炎病態に関与するStaphylococcus pseudintermedius分子の探索

    島倉 秀勝, 鶴井 大幹, 坂本 修士, 樋口 琢磨, 伊從 慶太, 下池 健太, 阪口 雅弘, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   157 - 157   2020.1

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  • 組換えendolysinのブドウ球菌に対する溶菌作用ならびに膿痂疹モデルマウスにおける発症抑制効果

    今西 市朗, 内山 淳平, 津久井 利広, 久恒 順三, 井手 香織, 松崎 茂展, 菅井 基行, 西藤 公司

    日本細菌学雑誌   75 ( 1 )   97 - 97   2020.1

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  • Clostridioides difficile感染症に対する糞便微生物移植治療についてのオミックス解析(Omics analysis in fecal transplantation therapy for Clostridioides difficile infection)

    阪口 義彦, 後藤 和義, 妹尾 充敏, 武 晃, 内山 淳平, 尾崎 隼人, 城代 康貴, 林 俊治, 大宮 直木, 加藤 はる

    日本細菌学雑誌   75 ( 1 )   125 - 125   2020.1

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  • Clostridioides difficile感染症に対する糞便微生物移植治療についてのオミックス解析(Omics analysis in fecal transplantation therapy for Clostridioides difficile infection)

    阪口 義彦, 後藤 和義, 妹尾 充敏, 武 晃, 内山 淳平, 尾崎 隼人, 城代 康貴, 林 俊治, 大宮 直木, 加藤 はる

    日本細菌学雑誌   75 ( 1 )   125 - 125   2020.1

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  • 牛白血病ウイルス感染による乳牛の腸内細菌叢の解析

    内山 淳平, 村上 裕信, 佐藤 礼一郎, 水上 圭二郎, 鈴木 武人, 島 綾香, 石原 玄基, 阪口 義彦, 曽川 一幸, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   126 - 126   2020.1

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  • アトピー性皮膚炎犬におけるワクチン副反応リスクの検討

    杉本 良輔, 島倉 秀勝, 内山 淳平, 津久井 利広, 藤村 正人, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   97 - 97   2020.1

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  • 組換えendolysinのブドウ球菌に対する溶菌作用ならびに膿痂疹モデルマウスにおける発症抑制効果

    今西 市朗, 内山 淳平, 津久井 利広, 久恒 順三, 井手 香織, 松崎 茂展, 菅井 基行, 西藤 公司

    日本細菌学雑誌   75 ( 1 )   97 - 97   2020.1

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  • イヌの純血コロニーを利用したアトピー性皮膚炎に関連した口腔・腸内細菌叢の解析

    海野 朝香, 今西 市朗, 水上 圭二郎, 大隅 尊史, 五十嵐 寛高, 村上 裕信, 島 綾香, 宇根 有美, 阪口 雅弘, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   76 - 76   2020.1

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  • アトピー性皮膚炎犬におけるワクチン副反応リスクの検討

    杉本 良輔, 島倉 秀勝, 内山 淳平, 津久井 利広, 藤村 正人, 阪口 雅弘

    日本細菌学雑誌   75 ( 1 )   97 - 97   2020.1

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  • グラム陽性細菌が産生する膜小胞の精製法の検討

    那須川 忠弥, 杉本 良輔, 島倉 秀勝, 小方 雅也, 福田 憲, 松崎 茂展, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   73 - 73   2020.1

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  • イヌの純血コロニーを利用したアトピー性皮膚炎に関連した口腔・腸内細菌叢の解析

    海野 朝香, 今西 市朗, 水上 圭二郎, 大隅 尊史, 五十嵐 寛高, 村上 裕信, 島 綾香, 宇根 有美, 阪口 雅弘, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   76 - 76   2020.1

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  • イヌにおけるアトピー性皮膚炎病態に関与するStaphylococcus pseudintermedius分子の探索

    島倉 秀勝, 鶴井 大幹, 坂本 修士, 樋口 琢磨, 伊從 慶太, 下池 健太, 阪口 雅弘, 内山 淳平

    日本細菌学雑誌   75 ( 1 )   157 - 157   2020.1

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  • Analysis of gut microbiome in dairy cows infected with bovine leukemia virus

    内山淳平, 村上裕信, 佐藤礼一郎, 水上圭二郎, 鈴木武人, 島綾香, 石原玄基, 阪口義彦, 阪口義彦, 曽川一幸, 阪口雅弘

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • 牛伝染性リンパ腫ウイルス感染による乳牛の腸内細菌叢への影響

    内山淳平, 村上裕信, 佐藤礼一郎, 水上圭二郎, 鈴木武人, 島綾香, 石原玄基, 曽川一幸, 阪口雅弘

    日本ワンヘルスサイエンス学会年次学術集会抄録集(Web)   4th   2020

  • Analysis of oral and gut microbiome associated with atopic dermatitis, using purebred dog colony

    海野朝香, 今西市朗, 水上圭二郎, 大隅尊史, 五十嵐寛高, 村上裕信, 島綾香, 宇根有美, 阪口雅弘, 内山淳平

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • Therapeutic potential of an endolysin derived from bacteriophage S25-3 for Staphylococcal impetigo

    今西市朗, 内山淳平, 津久井利広, 久恒順三, 井手香織, 松崎茂展, 菅井基行, 西藤公司

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • 複数の獣医系機関におけるイヌの膿皮症やアレルギー性皮膚炎に対する抗菌薬非依存的な取り組み

    内山淳平, 今西市朗, 今西市朗, 西藤公司, 福山朋季, 伊從慶太, 津久井利広, 阪口雅弘

    日本ブドウ球菌研究会プログラム・講演要旨集   65th (Web)   2020

  • バクテリオファージを利用した高選択的な細菌検出技術の開発

    今井斉志, 仁子陽輔, 波多野慎悟, 渡辺茂, 内山伊代, 内山淳平, 松崎茂展

    日本分析化学会年会講演要旨集(Web)   69th   2020

  • ネコ用ワクチン接種後のアナフィラキシーに関する研究

    吉田愛美, 吉田愛美, 伊藤雅人, 水上圭二郎, 水上圭二郎, 内山淳平, 阪口雅弘

    日本ワンヘルスサイエンス学会年次学術集会抄録集(Web)   4th   2020

  • Search for Staphylococcus pseudintermedius molecule associated with atopic dermatitis in dogs.

    島倉秀勝, 島倉秀勝, 鶴井大幹, 坂本修士, 樋口琢磨, 伊從慶太, 下池健太, 阪口雅弘, 内山淳平

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • Examination of adverse reaction risk to vaccine in atopic dermatitis dogs.

    杉本良輔, 島倉秀勝, 内山淳平, 津久井利広, 藤村正人, 阪口雅弘

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • Examination of the purification method of the membrane vesicles produced by Gram-positive bacteria

    那須川忠弥, 杉本良輔, 島倉秀勝, 小方雅也, 福田憲, 松崎茂展, 内山淳平

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • Omics analysis in fecal transplantation therapy for Clostridioides difficile infection

    阪口義彦, 後藤和義, 妹尾充敏, 武晃, 内山淳平, 尾崎隼人, 城代康貴, 林俊治, 大宮直木, 加藤はる

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • Development of a reagent to increase accuracy of GBS screening test for pregnant women

    小方雅也, 内山淳平, 松井秀仁, 内山伊代, 那須川忠弥, 松崎茂展, 花木秀明, 阪口雅弘

    日本細菌学雑誌(Web)   75 ( 1 )   2020

  • 新規神経細胞保護物質の探索及び作用機構解析

    鶴川 幸音, 菅野 和紀, 鈴木 優華, 村上 裕信, 内山 淳平, 竹田 志郎, 永根 大幹, 坂上 元栄, 紙透 伸治

    日本獣医学会学術集会講演要旨集   162回   485 - 485   2019.8

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  • 加齢によるイヌの腸内細菌叢の変化

    水上 圭二郎, 内山 淳平, 五十嵐 寛高, 村上 裕信, 大隅 尊史, 島 綾香, 石原 玄基, 那須川 忠弥, 宇根 有美, 阪口 雅弘

    日本獣医学会学術集会講演要旨集   162回   369 - 369   2019.8

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  • Current state of microbiome knowledge in dogs and cats Invited Reviewed

    五十嵐寛高, 内山淳平, 根尾櫻子, 村上裕信, 福山朋季, 久末正晴, 阪口雅弘

    月刊Precision Medicine   2 ( 4 )   373 - 377   2019.4

  • 一般遠心機を使用してウイルスを精製する

    那須川忠弥, 内山淳平, 田原口智士, 松崎茂展, 阪口雅弘

    麻布大学雑誌(Web)   30 ( 30 )   84 - 84   2019.3

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  • Clostridioides difficile感染症に対する糞便移植療法における腸管細菌叢と代謝産物の分析(Analysis of gut microbiota and metabolite in fecal transplantation therapy for Clostridioides difficile infection)

    阪口 義彦, 後藤 和義, 妹尾 充敏, 内山 淳平, 尾崎 隼人, 城代 康貴, 林 俊治, 大宮 直木, 加藤 はる

    日本細菌学雑誌   74 ( 1 )   109 - 109   2019.3

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  • B群連鎖球菌検査におけるファージ溶菌酵素の利用の可能性

    小方 雅也, 内山 淳平, 松井 秀仁, 松崎 茂展, 花木 秀明

    感染症学雑誌   93 ( 臨増 )   387 - 387   2019.3

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  • グループB連鎖球菌感染症の出生前スクリーニング法のファージを用いた改善可能性(Potential improvement of prenatal Group B Streptococcus screening using phages)

    内山 淳平, 松井 秀仁, 那須川 忠弥, 阪口 義彦, 水上 圭二郎, 阪口 雅弘, 松崎 茂展, 花木 秀明

    日本細菌学雑誌   74 ( 1 )   66 - 66   2019.3

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  • 犬アトピー性皮膚炎を悪化させるStaphylococcus pseudintermedius分子の探索

    那須川 忠弥, 内山 淳平, 鶴井 大幹, 坂本 修士, 樋口 琢磨, 伊從 慶太, 下池 健太, 島倉 秀勝, 松崎 茂展, 阪口 雅弘

    日本細菌学雑誌   74 ( 1 )   140 - 140   2019.3

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  • B群連鎖球菌検査におけるファージ溶菌酵素の利用の可能性

    小方雅也, 内山淳平, 松井秀仁, 松崎茂展, 花木秀明

    感染症学雑誌   93 ( 臨増 )   387 - 387   2019.3

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  • ロボショットにおけるIoTへの対応—Roboshot's Response to IoT—特集 新しいモノづくりへの挑戦

    内山 辰宏, 丸山 淳平, 浅岡 裕泰

    プラスチックスエージ = Plastics age   65 ( 1 )   62 - 67   2019.1

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  • 純血種犬コロニーを用いた腸内細菌叢の加齢性変化の同定

    水上圭二郎, 水上圭二郎, 内山淳平, 五十嵐寛高, 村上裕信, 大隅尊史, 島綾香, 島綾香, 石原玄基, 那須川忠弥, 宇根有美, 宇根有美, 阪口雅弘

    日本ワンヘルスサイエンス学会年次学術集会抄録集(Web)   3rd   2019

  • Search for Staphylococcus pseudintermedius molecules exacerbating canine atopic dermatitis

    那須川忠弥, 内山淳平, 鶴井大幹, 坂本修士, 樋口琢磨, 伊從慶太, 下池健太, 島倉秀勝, 松崎茂展, 阪口雅弘

    日本細菌学雑誌(Web)   74 ( 1 )   2019

  • Analysis of gut microbiota and metabolite in fecal transplantation therapy for Clostridioides difficile infection

    阪口義彦, 後藤和義, 妹尾充敏, 内山淳平, 尾崎隼人, 城代康貴, 林俊治, 大宮直木, 加藤はる

    日本細菌学雑誌(Web)   74 ( 1 )   2019

  • イヌの狂犬病ワクチン接種後のアナフィラキシーに関する研究

    吉田愛美, 伊藤雅人, 楠見夏樹, 水上圭二郎, 水上圭二郎, 内山淳平, 阪口雅弘

    日本ワンヘルスサイエンス学会年次学術集会抄録集(Web)   3rd   2019

  • イヌの混合ワクチン接種後アナフィラキシーに関する研究:ヒトの予防接種後副反応の標準化症例定義(Brighton分類)の応用

    伊藤雅人, 水上圭二郎, 水上圭二郎, 内山淳平, 阪口雅弘

    日本ワンヘルスサイエンス学会年次学術集会抄録集(Web)   3rd   2019

  • 出産前B群連鎖球菌培養検査における検査精度向上のためのファージ利用の可能性

    内山 淳平, 松井 秀仁, 花木 秀明

    日本臨床微生物学会雑誌   29 ( Suppl.1 )   368 - 368   2018.12

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  • Pestalotiopsis microsporaが生産する神経細胞保護活性物質の単離と構造決定

    菅野和紀, 鶴川幸音, 柴崎久宣, 村上裕信, 内山淳平, 倉持幸司, 紙透伸治

    日本農芸化学会関東支部講演要旨集   2018 ( Oct )   18   2018.10

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  • ウイルスの系統とゲノム組換え―Earth’s viromeデータの解析から― Invited International journal

    矢原耕史, MEIER‐KOLTHOFF Jan P, 内山淳平, 矢原寛子, PAEZ‐ESPINO David

    日本進化学会大会プログラム・講演要旨集(Web)   20th ( 1 )   87 (WEB ONLY) - 11496   2018.8

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    Bacteriophages (phages), or bacterial viruses, are the most abundant and diverse biological entities that impact the global ecosystem. Recent advances in metagenomics have revealed their rampant abundance in the biosphere. A fundamental aspect of bacteriophages that remains unexplored in metagenomic data is the process of recombination as a driving force in evolution that occurs among different viruses within the same bacterial host. Here, we systematically examined signatures of recombination in every gene from 211 species-level viral groups in a recently obtained dataset of the Earth’s virome that contain corresponding information on the host bacterial species. Our study revealed that signatures of recombination are widespread (84%) among the diverse viral groups. We identified 25 recombination-intense viral groups, widely distributed across the viral taxonomy, and present in bacterial species living in the human oral cavity. We also revealed a significant inverse association between the recombination-intense viral groups and Type II restriction endonucleases, that could be effective in reducing recombination among phages in a cell. Furthermore, we identified recombination-intense genes that are significantly enriched for encoding phage morphogenesis proteins. Changes in the viral genomic sequence by recombination may be important to escape cleavage by the host bacterial immune systems.

    DOI: 10.1038/s41598-018-29272-2

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  • 牛白血病ウイルス野外株の遺伝的多様性と病原性への影響

    村上裕信, 河合将克, 佐籐礼一郎, 前田洋佑, 加藤肇, 内山淳平, 阪口雅弘, 塚本健司

    日本獣医学会学術集会講演要旨集   161回   352 - 352   2018.8

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  • 臨床皮膚科でいま“できる”ことを提案する 第3回 膿皮症アップデート―夏の強敵と対峙する!―CCG6 メチシリン耐性ブドウ球菌の同定

    内山淳平

    Small Animal Dermatology   14 ( 4 )   19‐25 - 25   2018.7

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  • 動物における「マイクロバイオーム」研究の現状と応用 マイクロバイオーム解析の基礎 細菌叢解析を中心に Invited

    内山 淳平

    ペット栄養学会誌   21 ( 2 )   S11 - S11   2018.7

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  • 【臨床皮膚科でいま"できる"ことを提案する 第3回 膿皮症アップデート-夏の強敵と対峙する!-】 ヒトの医療におけるメチシリン耐性黄色ブドウ球菌の除菌

    内山 淳平

    Small Animal Dermatology   14 ( 4 )   34 - 35   2018.7

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  • 臨床皮膚科でいま“できる”ことを提案する 第3回 膿皮症アップデート―夏の強敵と対峙する!―CCG10 ヒトの医療におけるメチシリン耐性黄色ブドウ球菌の除菌

    内山淳平

    Small Animal Dermatology   14 ( 4 )   34 - 35   2018.7

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  • CCG6 メチシリン耐性ブドウ球菌の同定 (特集 臨床皮膚科でいま"できる"ことを提案する(第3回)膿皮症アップデート 夏の強敵と対峙する!)

    内山 淳平

    Small animal dermatology : 小動物皮膚科専門誌   52 ( 4 )   19 - 25   2018.7

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    Other Link: http://search.jamas.or.jp/link/ui/2018300752

  • バクテリオファージを利用する多剤耐性細菌感染症制御法 Invited Reviewed

    松﨑茂展, 内山淳平

    動物用ワクチン・バイオ医薬品研究会 NEWSletter   17 ( 17 )   11 - 16   2018.6

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  • バクテリオファージ担持金ナノ粒子凝集体を利用した細菌の暗視野顕微鏡検出

    今井斉志, 鷲尾和也, 仁子陽輔, 波多野慎悟, 渡辺茂, 松崎茂展, 内山淳平

    日本化学会春季年会講演予稿集(CD-ROM)   98th   ROMBUNNO.3G3‐02   2018.3

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  • C型とD型ボツリヌス毒素変換ファージの解析(Analysis of botulinum neurotoxin-converting phages in Clostridium botulinum types C and D)

    阪口 義彦, 内山 淳平, 小椋 義俊, 後藤 和義, 山本 由弥子, 松崎 茂展, 山口 明日美, 林 哲也, 小熊 惠二, 林 俊治

    日本細菌学雑誌   73 ( 1 )   94 - 94   2018.2

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  • Acinetobacter baumanniiの同定方法の比較検討

    山口 明日美, 阪口 義彦, 松井 真理, 内山 淳平, 松井 秀仁, 花木 秀明, 小林 秀丈, 小山内 洋子, 林 俊治

    日本細菌学雑誌   73 ( 1 )   66 - 66   2018.2

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  • 関東の医療施設で分離されたAcinetobacter baumanniiの分子疫学的解析

    山口 明日美, 阪口 義彦, 松井 真理, 小林 秀丈, 内山 淳平, 小山内 洋子, 松井 秀仁, 花木 秀明, 林 俊治

    日本細菌学雑誌   73 ( 1 )   52 - 52   2018.2

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  • C型とD型ボツリヌス毒素変換ファージの解析(Analysis of botulinum neurotoxin-converting phages in Clostridium botulinum types C and D)

    阪口 義彦, 内山 淳平, 小椋 義俊, 後藤 和義, 山本 由弥子, 松崎 茂展, 山口 明日美, 林 哲也, 小熊 惠二, 林 俊治

    日本細菌学雑誌   73 ( 1 )   94 - 94   2018.2

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  • Acinetobacter baumanniiの同定方法の比較検討

    山口 明日美, 阪口 義彦, 松井 真理, 内山 淳平, 松井 秀仁, 花木 秀明, 小林 秀丈, 小山内 洋子, 林 俊治

    日本細菌学雑誌   73 ( 1 )   66 - 66   2018.2

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  • 関東の医療施設で分離されたAcinetobacter baumanniiの分子疫学的解析

    山口 明日美, 阪口 義彦, 松井 真理, 小林 秀丈, 内山 淳平, 小山内 洋子, 松井 秀仁, 花木 秀明, 林 俊治

    日本細菌学雑誌   73 ( 1 )   52 - 52   2018.2

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  • 関東の医療施設で分離されたAcinetobacter baumanniiの分子疫学的解析

    山口明日美, 阪口義彦, 松井真理, 小林秀丈, 内山淳平, 小山内洋子, 松井秀仁, 花木秀明, 林俊治

    日本細菌学雑誌(Web)   73 ( 1 )   2018

  • Introduction to gut microflora analysis

    Uchiyama Jumpei

    Journal of Pet Animal Nutrition   21 ( 3 )   142 - 144   2018

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    DOI: 10.11266/jpan.21.3_142

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  • 腸球菌眼内炎に対するバクテリオファージ療法の効果

    岸本達真, 石田わか, 鈴木崇, 内山淳平, 松崎茂展, 大畑雅典, 福田憲, 福島敦樹

    日本眼感染症学会・日本眼炎症学会・日本コンタクトレンズ学会総会・日本涙道・涙液学会プログラム・講演抄録集   55th-52nd-61st-7th   69   2018

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  • Acinetobacter baumanniiのresistance islandの遺伝子解析

    山口 明日美, 松井 真理, 内山 淳平, 松井 秀仁, 花木 秀明, 林 俊治, 阪口 義彦, 小山内 洋子, 小林 秀丈

    日本臨床微生物学雑誌   28 ( Suppl.1 )   490 - 490   2017.12

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  • Acinetobacter baumanniiのresistance islandの遺伝子解析

    山口 明日美, 松井 真理, 内山 淳平, 松井 秀仁, 花木 秀明, 林 俊治, 阪口 義彦, 小山内 洋子, 小林 秀丈

    日本臨床微生物学雑誌   28 ( Suppl.1 )   490 - 490   2017.12

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  • Acinetobacter baumanniiの同定方法の比較検討

    山口 明日美, 阪口 義彦, 小山内 洋子, 林 俊治, 松井 秀仁, 花木 秀明, 松井 真理, 内山 淳平

    首都圏支部・関甲信支部医学検査学会プログラム・講演抄録集   54回 ( 1 )   150 - 150   2017.10

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  • 【視野を広げる皮膚科診療のTips(秘訣)第10回 皮膚科のお悩み相談室】膿皮症 膿皮症の診断・治療に際して、なかなか細菌検査を実施することができません。検査の重要性、実施法や検査機関の選択などを教えてください

    内山 淳平, 阪口 雅弘, 加藤 行男, 伊從 慶太

    Small Animal Dermatology   13 ( 4 )   17 - 26   2017.7

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  • Q 膿皮症の診断・治療に際して,なかなか細菌検査を実施することができません。検査の重要性,実施法や検査機関の選択などを教えてください。 (特集 視野を広げる皮膚科診療のTips(秘訣)(第10回)皮膚科の"お悩み"相談室) -- (膿皮症)

    内山 淳平, 阪口 雅弘, 加藤 行男, 伊從 慶太

    Small animal dermatology : 小動物皮膚科専門誌   46   17 - 26   2017.7

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    Other Link: http://search.jamas.or.jp/link/ui/2017336729

  • 視野を広げる皮膚科診療のTips(秘訣)第10回 皮膚科の“お悩み”相談室 II・膿皮症 Q6 膿皮症の診断・治療に際して,なかなか細菌検査を実施することができません。検査の重要性,実施法や検査機関の選択などを教えてください。

    内山淳平, 阪口雅弘, 加藤行男, 伊從慶太

    Small Animal Dermatology   46 ( 4 )   17 - 26   2017.7

  • 組換えendolysinのブドウ球菌に対する溶菌作用ならびに膿痂疹モデルマウスにおける発症抑制効果

    今西市朗, 内山淳平, 水谷歩実, 井手香織, 西藤公司

    日本獣医皮膚科学会学術大会・総会   20th   88   2017.3

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  • バクテリオファージKHP30の感染能に及ぼすピロリ菌保有制限 修飾系の解析

    松澤 佑一, 内山 淳平, 竹内 啓晃, 氏原 隆子, 橋田 裕美子, 樋口 智紀, 田中 望紅, 冨永 宗一竜, 大畑 雅典, 松崎 茂展

    日本細菌学雑誌   72 ( 1 )   95 - 95   2017.2

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  • 黄色ブドウ球菌に感染するAHJD様ファージの吸着に関する研究(Study of adsorption of AHJD-like phages infecting Staphylococcus aureus)

    内山 淳平, 黒川 健児, 内山 伊代, 氏原 隆子, 阪口 義彦, 松崎 茂展, 阪口 雅弘

    日本細菌学雑誌   72 ( 1 )   169 - 169   2017.2

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  • バクテリオファージKHP30の感染能に及ぼすピロリ菌保有制限 修飾系の解析

    松澤 佑一, 内山 淳平, 竹内 啓晃, 氏原 隆子, 橋田 裕美子, 樋口 智紀, 田中 望紅, 冨永 宗一竜, 大畑 雅典, 松崎 茂展

    日本細菌学雑誌   72 ( 1 )   95 - 95   2017.2

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  • B6型ボツリヌス毒素遺伝子をコードするプラスミドの解析(Analysis of botulinum neurotoxin type B6 gene-encoding plasmid in Clostridium botulinum type B)

    阪口 義彦, 内山 淳平, 細見 晃司, 幸田 知子, 小椋 義俊, 林 哲也, 林 俊治, 小崎 俊司, 向本 雅郁

    日本細菌学雑誌   72 ( 1 )   127 - 127   2017.2

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  • B6型ボツリヌス毒素遺伝子をコードするプラスミドの解析(Analysis of botulinum neurotoxin type B6 gene-encoding plasmid in Clostridium botulinum type B)

    阪口 義彦, 内山 淳平, 細見 晃司, 幸田 知子, 小椋 義俊, 林 哲也, 林 俊治, 小崎 俊司, 向本 雅郁

    日本細菌学雑誌   72 ( 1 )   127 - 127   2017.2

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  • 黄色ブドウ球菌に感染するAHJD様ファージの吸着に関する研究(Study of adsorption of AHJD-like phages infecting Staphylococcus aureus)

    内山 淳平, 黒川 健児, 内山 伊代, 氏原 隆子, 阪口 義彦, 松崎 茂展, 阪口 雅弘

    日本細菌学雑誌   72 ( 1 )   169 - 169   2017.2

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  • バクテリオファージKHP30の感染能に及ぼすピロリ菌保有制限-修飾系の解析

    松澤佑一, 内山淳平, 竹内啓晃, 氏原隆子, 橋田裕美子, 樋口智紀, 田中望紅, 冨永宗一竜, 大畑雅典, 松崎茂展

    日本細菌学雑誌(Web)   72 ( 1 )   2017

  • Helicobacter pylori ファージとそのゲノム (特集 Helicobacter pyloriのゲノムサイエンス)

    内山 淳平, 竹内 啓晃, 平山 隆一郎, 島倉 秀勝, 阪口 雅弘, 松崎 茂展

    Helicobacter research : Journal of helicobacter research   20 ( 5 )   470 - 474   2016.10

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    筆者らはHelicobacter pyloriに感染する新規バクテリオファージ(ファージ)KHP30を分離・解析し、そのユニークな性状・ゲノムを明らかにした。近年、KHP30に遺伝的に類似した3種のKHP30様ファージが報告されている。本稿では、ファージKHP30の特徴を概説し、KHP30様ファージゲノムに関して解説する。(著者抄録)

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  • Helicobacterのゲノムサイエンス Helicobacter pyloriファージとそのゲノム

    内山淳平, 竹内啓晃, 平山隆一郎, 島倉秀勝, 阪口雅弘, 松崎茂展

    Helicobacter Research   20 ( 5 )   470 - 474   2016.10

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    筆者らはHelicobacter pyloriに感染する新規バクテリオファージ(ファージ)KHP30を分離・解析し、そのユニークな性状・ゲノムを明らかにした。近年、KHP30に遺伝的に類似した3種のKHP30様ファージが報告されている。本稿では、ファージKHP30の特徴を概説し、KHP30様ファージゲノムに関して解説する。(著者抄録)

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  • ファージ・ルネッサンス ファージ発見から100年たった今 活性型ピロリ菌ファージKHP30の特徴付けとその潜伏感染性の解析(Phage renaissance: 100 years have passed since phage discovery Characterization of active Helicobacter pylori phage KHP30, and analysis of its latent infection)

    内山 淳平, 内山 伊代, 竹内 啓晃, 阪口 義彦, 阪口 雅弘, 松崎 茂展

    日本細菌学雑誌   71 ( 1 )   53 - 53   2016.2

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  • C型ボツリヌス毒素変換ファージのゲノム比較と遺伝子機能解析(The genome sequence of Clostridium botulinum type C phage and functional analysis of gene products)

    阪口 義彦, 内山 淳平, 小椋 義俊, 山本 由弥子, 松崎 茂展, 林 哲也, 松下 治, 小熊 惠二, 林 俊治

    日本細菌学雑誌   71 ( 1 )   135 - 135   2016.2

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  • ファージ・ルネッサンス ファージ発見から100年たった今 ファージを利用した細菌検出 黄色ブドウ球菌検出ファージクロマト法の開発

    松井 秀仁, 内山 淳平, 津田 愛美, 松崎 茂展, 花木 秀明

    日本細菌学雑誌   71 ( 1 )   54 - 54   2016.2

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  • 緑膿菌PAO1株とファージKPP22の短期間進化的軍拡競走の解析

    那須川 忠弥, 内山 淳平, 鈴木 仁人, 宮田 玲奈, 山口 琴絵, 内山 伊代, 阪口 義彦, 柴山 恵吾, 阪口 雅弘, 松崎 茂展

    日本細菌学雑誌   71 ( 1 )   92 - 92   2016.2

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  • Helicobacter pylori ファージとそのゲノム

    内山 淳平, 竹内 啓晃, 松崎 茂展

    Helicobacter Research   20   50 - 54   2016

  • 食物有害反応を示す犬における卵白アレルゲンに対するIgE反応性の検討

    島倉秀勝, 齋藤拓, 藤村正人, 岡本憲明, 内山淳平, 阪口雅弘

    日本獣医学会学術集会講演要旨集   158回   401 - 401   2015.8

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  • 細菌感染症における診断法と治療法の開発:バクテリオファージの利用

    内山淳平, 松井秀仁, 阪口義彦, 花木秀明, 松崎茂展, 阪口雅弘

    日本獣医学会学術集会講演要旨集   158回   182 - 182   2015.8

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  • イヌにおける新規口腔内常在細菌の同定と全ゲノム配列解析

    平山隆一郎, 平山隆一郎, 鈴木仁人, 内山淳平, 松井真理, 鈴木里和, 柴山恵吾, 阪口雅弘, 木内明男

    日本獣医学会学術集会講演要旨集   158回   313 - 313   2015.8

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  • 眼感染症に対するファージ療法の研究開発および臨床応用への問題点

    福田憲, 石田わか, 内山淳平, 松崎茂展, 福島敦樹

    日本獣医学会学術集会講演要旨集   158回   183 - 183   2015.8

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  • Evaluation of the lubrication mechanism at various rotation speeds and granule filling levels in a container mixer using a thermal effusivity sensor Reviewed International journal

    Jumpei Uchiyama, Shigeru Aoki

    DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY   41 ( 7 )   1172 - 1181   2015.7

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    To research the detailed mechanism of the lubrication process using the thermal effusivity sensor, the relationships of the lubrication progress with the pattern of powder flow, the rotation speed and the filling level were investigated. The thermal effusivity profile was studied as a function of the number of rotations at various rotation speeds. It was observed that at lower rotation speeds, the profiles of the lubrication progress were almost the same, regardless of the rotation speed. In this region, the highest speed was defined as the critical rotation speed (CRS), which was found to be one of the important factors. The CRS had close relations with avalanche flow in the blender. The first and the second phases were observed in the lubrication process. The first phase was influenced by the CRS and the filling level in the blender. The second phase was influenced by the rotation speed. The mechanism of two-phase process was proposed as a macro progression of the dispersion of the lubricant (first phase) and micro progression of the coating of the powder particles with lubricant (second phase). The accurate monitoring by the thermal effusivity sensor was able to help a better understanding in the lubrication process.

    DOI: 10.3109/03639045.2014.935394

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  • ファージ尾部タンパクを用いた黄色ブドウ球菌検出方法の開発

    松井 秀仁, 崔 龍洙, 花木 秀明, 内山 淳平, 松崎 茂展

    日本化学療法学会雑誌   63 ( Suppl.A )   170 - 170   2015.5

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  • New Approach to Evaluate the Lubrication Process in Various Granule Filling Levels and Rotating Mixer Sizes Using a Thermal Effusivity Sensor Reviewed

    Jumpei Uchiyama, Shigeru Aoki, Yoshifumi Uemoto

    CHEMICAL & PHARMACEUTICAL BULLETIN   63 ( 3 )   164 - 179   2015.3

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    The principles of thermal effusivity are applied to an understanding of the detailed mechanisms of the lubrication process in a rotating mixer. The relationships and impact of the lubrication process by the pattern of powder flow, the filling level, and the rotating mixer size were investigated. Thermal effusivity profiles of the lubrication process, as obtained, indicate that lubrication is a two-phase process. The intersection point of the first and second phases (IPFS) is influenced by changing the filling level, thus changing the resulting number of avalanche flows created. The slope of the second phase (SSP) is influenced by the relationship between the number and the length of avalanche flows. Understanding this difference between the first and second phases is important to successfully evaluate the impact of proposed changes in the lubrication process. From this knowledge, a predictive model of the lubrication profile can be generated to allow an evaluation of proposed changes to the lubrication process. This model allows estimation of the lubrication profile at different filling levels and in different rotating mixer sizes. In this study, the actual lubrication profile almost coincides with the model predicted lubrication profile. Based on these findings, it is assumed that lubrication profiles at a commercial scale can be predicted from data generated at the laboratory scale. Further, it is assumed that changes in the filling level can also be estimated from the laboratory or current data.

    DOI: 10.1248/cpb.c14-00634

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  • 各種ファージに対する黄色ブドウ球菌受容体の解析

    内山淳平, 黒川健児, 坂口義彦, 内山(竹村)伊代, 氏原隆子, 村上雅尚, 大畑雅典, 松崎茂展

    日本細菌学雑誌   70 ( 1 )   215 - 215   2015.2

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  • 緑膿菌感染症に対するファージ療法と化学療法の併用療法の可能性検討

    重久立, 内山淳平, 宮田怜奈, 山口琴絵, 内山伊代, 阪口義彦, 氏原隆子, 大畑雅典, 松崎茂展

    日本細菌学雑誌   70 ( 1 )   198 - 198   2015.2

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  • B型ボツリヌス菌のボツリヌス毒素遺伝子をコードするプラスミド解析(Analysis of botulinum neurotoxin type B6 gene-encoding plasmid in Clostridium botulinum type B)

    阪口 義彦, 内山 淳平, 細見 晃司, 幸田 知子, 松崎 茂展, 小椋 義俊, 林 哲也, 林 俊治, 小崎 俊司, 向本 雅郁

    日本細菌学雑誌   70 ( 1 )   178 - 178   2015.2

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  • クライオ電子顕微鏡による新規大型黄色ブドウ球菌バクテリオファージS6の構造解析

    宮崎直幸, 内山淳平, 松崎茂展, 村田和義

    日本生化学会大会(Web)   87回   [2P - 130]   2014.10

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  • Genomic and phylogenetic traits of Staphylococcus phages S25-3 and S25-4 (family Myoviridae, genus Twort-like viruses) Reviewed

    Iyo Takemura-Uchiyama, Jumpei Uchiyama, Shin Ichiro Kato, Takako Ujihara, Masanori Daibata, Shigenobu Matsuzaki

    Annals of Microbiology   64 ( 3 )   1453 - 1456   2014.9

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    The genomes of Staphylococcus phages S25-3 and S25-4 (family Myoviridae, genus Twort-like viruses) were sequenced and analyzed from an evolutionary perspective. The genome-based phylogeny and genome analyses of phages S25-3 and S25-4 showed that they had diverged evolutionarily from the majority of this viral genus based on the presence of mobile genetic elements, i.e., a putative transposase and the homing endonuclease I-MsaI. These results suggest that genetic elements such as transposases and homing endonucleases are likely to be involved with the evolution of some Twort-like phages, including phages S25-3 and S25-4. © 2013 Springer-Verlag Berlin Heidelberg and the University of Milan.

    DOI: 10.1007/s13213-013-0762-2

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  • 最先端医療・福祉の科学 ファージ吸着分子を利用した簡易迅速細菌検出技術

    内山淳平, 松井秀仁, 内山(竹村)伊代, 渡辺茂, 花木秀明, 松崎茂展

    化学工業   65 ( 8 )   588 - 595   2014.8

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  • Bacteria Detection Technology Using Phage Molecule

    内山 淳平, 松井 秀仁, 内山(竹村) 伊代

    化學工業   65 ( 8 )   588 - 595   2014.8

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  • ファージ吸着分子を利用した簡易迅速細菌検査技術 Invited

    内山淳平, 松井秀仁, 内山(竹村)伊代, 渡辺茂, 花木秀明, 松﨑茂展

    化学工業   65 ( 8 )   12 - 19   2014.8

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  • Evaluation of risk and benefit in thermal effusivity sensor for monitoring lubrication process in pharmaceutical product manufacturing Reviewed

    Jumpei Uchiyama, Yoshiteru Kato, Yoshifumi Uemoto

    DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY   40 ( 8 )   999 - 1004   2014.8

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    In the process design of tablet manufacturing, understanding and control of the lubrication process is important from various viewpoints. A detailed analysis of thermal effusivity data in the lubrication process was conducted in this study. In addition, we evaluated the risk and benefit in the lubrication process by a detailed investigation. It was found that monitoring of thermal effusivity detected mainly the physical change of bulk density, which was changed by dispersal of the lubricant and the coating powder particle by the lubricant. The monitoring of thermal effusivity was almost the monitoring of bulk density, thermal effusivity could have a high correlation with tablet hardness. Moreover, as thermal effusivity sensor could detect not only the change of the conventional bulk density but also the fractional change of thermal conductivity and thermal capacity, two-phase progress of lubrication process could be revealed. However, each contribution of density, thermal conductivity, or heat capacity to thermal effusivity has the risk of fluctuation by formulation. After carefully considering the change factor with the risk to be changed by formulation, thermal effusivity sensor can be a useful tool for monitoring as process analytical technology, estimating tablet hardness and investigating the detailed mechanism of the lubrication process.

    DOI: 10.3109/03639045.2013.795581

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  • Development of new punch shape to replicate scale-up issues in laboratory tablet press II: a new design of punch head to emulate consolidation and dwell times in commercial tablet press. Reviewed International journal

    Shigeru Aoki, Jumpei Uchiyama, Manabu Ito

    Journal of pharmaceutical sciences   103 ( 6 )   1921 - 7   2014.6

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    Differences between laboratory and commercial tablet presses are frequently observed during scale-up of tableting process. These scale-up issues result from the differences in total compression time that is the sum of consolidation and dwell times. When a lubricated blend is compressed into tablets, the tablet thickness produced by the commercial tablet press is often thicker than that by a laboratory tablet press. A new punch shape design, designated as shape adjusted for scale-up (SAS), was developed and used to demonstrate the ability to replicate scale-up issues in commercial-scale tableting processes. It was found that the consolidation time can be slightly shortened by changing the vertical curvature of the conventional punch head rim. However, this approach is not enough to replicate the consolidation time. A secondary two-stage SAS punch design and an embossed punch head was designed to replicate the consolidation and dwell times on a laboratory tablet press to match those of a commercial tablet press. The resulting tablet thickness using this second SAS punch on a laboratory tablet press was thicker than when using a conventional punch in the same laboratory tablet press. The secondary SAS punches are more useful tools for replicating and understanding potential scale-up issues. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci.

    DOI: 10.1002/jps.23989

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  • Examination of generalized transduction among Staphylococcus spp., using a novel phage S6

    UCHIYAMA JUMPEI, TAKEMURA-UCHIYAMA IYO, SAKAGUCHI YOSHIHIKO, MISAWA NAOAKI, DAIBATA MASANORI, MATSUZAKI SHIGENOBU

    日本細菌学雑誌   69 ( 1 )   236   2014.2

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  • 緑膿菌ファージKPP21,KPP22,KPP23,KPP25の網羅的構造タンパク質解析

    重久立, 内山淳平, 宮田怜奈, 山口琴絵, 内山伊代, 氏原隆子, 大畑雅典, 松崎茂展

    日本細菌学雑誌   69 ( 1 )   139 - 139   2014.2

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  • 緑膿菌ファージKPP22吸着能変異ファージの解析

    宮田怜奈, 内山淳平, 山口琴絵, 重久立, 内山伊代, 氏原隆子, 大畑雅典, 松崎茂展

    日本細菌学雑誌   69 ( 1 )   138 - 138   2014.2

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  • Phage therapy experiment against staphylococcal lung-derived septic mouse model

    UCHIYAMA IYO, UCHIYAMA JUMPEI, SAKAGUCHI YOSHIHIKO, UJIHARA TAKAKO, DAIBATA MASANORI, MATSUZAKI SHIGENOBU

    日本細菌学雑誌   69 ( 1 )   139   2014.2

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  • 緑膿菌ファージの分離とそれらのゲノム塩基配列

    山口琴絵, 内山淳平, 宮田怜奈, 重久立, 内山伊代, 氏原隆子, 大畑雅典, 松崎茂展

    日本細菌学雑誌   69 ( 1 )   138 - 138   2014.2

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  • Estimation of Helicobacter pylori KHP30-like phage ecology

    MATSUZAKI SHIGENOBU, UCHIYAMA JUMPEI, TAKEUCHI HIROAKI, SAKAGUCHI YOSHIHIKO, UCHIYAMA-TAKEMURA IYO, UJIHARA TAKAKO, DAIBATA MASANORI

    日本細菌学雑誌   69 ( 1 )   237   2014.2

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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(Examination of generalized transduction among Staphylococcus spp., using a novel phage S6)

    内山 淳平, 内山 伊代, 阪口 義彦, 三澤 尚明, 大畑 雅典, 松崎 茂展

    日本細菌学雑誌   69 ( 1 )   236 - 236   2014.2

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  • ピロリ菌におけるKHP30様ファージの存在様式(Estimation of Helicobacter pylori KHP30-like phage ecology)

    松崎 茂展, 内山 淳平, 竹内 啓晃, 阪口 義彦, 内山 伊代[竹村], 氏原 隆子, 大畑 雅典

    日本細菌学雑誌   69 ( 1 )   237 - 237   2014.2

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  • Clostridium argentinenseにおけるG型ボツリヌス神経毒素遺伝子をコードするプラスミドの分析(Analysis of botulinum neurotoxin type G gene-encoding plasmid in Clostridium argentinense)

    阪口 義彦, 内山 淳平, 鈴木 智典, 山本 由弥子, 小椋 義俊, 細見 晃司, 松崎 茂展, 林 哲也, 小崎 俊司, 小熊 惠二

    日本細菌学雑誌   69 ( 1 )   236 - 236   2014.2

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  • Clostridium argentinenseにおけるG型ボツリヌス神経毒素遺伝子をコードするプラスミドの分析(Analysis of botulinum neurotoxin type G gene-encoding plasmid in Clostridium argentinense)

    阪口 義彦, 内山 淳平, 鈴木 智典, 山本 由弥子, 小椋 義俊, 細見 晃司, 松崎 茂展, 林 哲也, 小崎 俊司, 小熊 惠二

    日本細菌学雑誌   69 ( 1 )   236 - 236   2014.2

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  • 黄色ブドウ球菌性肺由来敗血症マウスモデルにおけるファージ療法の検討(Phage therapy experiment against staphylococcal lungderived septic mouse model)

    内山 伊代, 内山 淳平, 阪口 義彦, 氏原 隆子, 大畑 雅典, 松崎 茂展

    日本細菌学雑誌   69 ( 1 )   139 - 139   2014.2

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  • A novel approach to a fine particle coating using porous spherical silica as core particles

    Makoto Ishida, Jumpei Uchiyama, Keiko Isaji, Yuta Suzuki, Yasuyuki Ikematsu, Shigeru Aoki

    Drug Development and Industrial Pharmacy   40   1054 - 1064   2014.1

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    The applicability of porous spherical silica (PSS) was evaluated as core particles for pharmaceutical products by comparing it with commercial core particles such as mannitol (NP-108), sucrose and microcrystalline cellulose spheres. We investigated the physical properties of core particles, such as particle size distribution, flow properties, crushing strength, plastic limit, drying rate, hygroscopic property and aggregation degree. It was found that PSS was a core particle of small particle size, low friability, high water adsorption capacity, rapid drying rate and lower occurrence of particle aggregation, although wettability is a factor to be carefully considered. The aggregation and taste-masking ability using PSS and NP-108 as core particles were evaluated at a fluidized-bed coating process. The functional coating under the excess spray rate shows different aggregation trends and dissolution profiles between PSS and NP-108; thereby, exhibiting the formation of uniform coating under the excess spray rate in the case of PSS. This expands the range of the acceptable spray feed rates to coat fine particles, and indicates the possibility of decreasing the coating time. The results obtained in this study suggested that the core particle, which has a property like that of PSS, was useful in overcoming such disadvantages as large particle size, which feels gritty in oral cavity; particle aggregation; and the long coating time of the particle coating process. These results will enable the practical fine particle coating method by increasing the range of optimum coating conditions and decreasing the coating time in fluidized bed technology. © 2014 Informa Healthcare USA, Inc.

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  • Development of new shaped punch to predict scale-up issue in tableting process. Reviewed International journal

    Shigeru Aoki, Jumpei Uchiyama, Manabu Ito

    Journal of pharmaceutical sciences   103 ( 1 )   235 - 40   2014.1

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    Scale-up issues in the tableting process, such as capping, sticking, or differences in tablet thickness, are often observed at the commercial production scale. A new shaped punch, named the size adjusted for scale-up (SAS) punch, was created to estimate scale-up issues seen between laboratory scale and commercial scale tableting processes. The SAS punch's head shape was designed to replicate the total compression time of a laboratory tableting machine to that of a commercial tableting machine. Three different lubricated blends were compressed into tablets using a laboratory tableting machine equipped with SAS punches, and any differences in tablet thickness or capping phenomenon were observed. It was found that the new shaped punch could be used to replicate scale-up issues observed in the commercial tableting machine. The SAS punch was shown to be a useful tool to estimate scale-up issues in the tableting process.

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  • Draft genome sequence of Clostridium botulinum type B strain Osaka05, isolated from an infant patient with botulism in Japan Reviewed

    Yoshihiko Sakaguchi, Koji Hosomi, Jumpei Uchiyama, Yoshitoshi Ogura, Kaoru Umeda, Masakiyo Sakaguchi, Tomoko Kohda, Masafumi Mukamoto, Naoaki Misawa, Shigenobu Matsuzaki, Tetsuya Hayashi, Shunji Kozaki

    Genome Announcements   2 ( 1 )   2014

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    © 2014 Sakaguchi et al. Clostridium botulinum strain Osaka05, which has been isolated from an infant patient with botulism in Japan, is the first strain producing botulinum neurotoxin subtype B6. Here, we report the draft genome sequence of C. botulinum Osaka05.

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  • Helicobacter pylori基礎研究の最前線 2.病原因子 Helicobacter pyloriのファージに関する基礎摘研究

    竹内啓晃, 内山淳平, 松崎茂展, 森本徳仁, 杉浦哲朗

    Helicobacter Res   17 ( 5 )   400 - 405   2013.10

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  • Helicobacter pylori ファージに関する基礎的研究 Invited

    竹内啓晃, 内山淳平, 松﨑茂展, 森本徳仁, 杉浦哲朗

    Helicobacter Research   17 ( 5 )   28 - 33   2013.10

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  • 【Helicobacter pylori基礎研究の最前線】 病原因子 Helicobacter pyloriのファージに関する基礎的研究

    竹内 啓晃, 内山 淳平, 松崎 茂展, 森本 徳仁, 杉浦 哲朗

    Helicobacter Research   17 ( 5 )   400 - 405   2013.10

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    尾部のないファージの分離率はわずか約4%とまれである。そのなかに球形ファージを含むさまざまな分類がある。われわれはHelicobacter pylori(H.pylori)が自然放出している二つの球形ファージ(KHP30とKHP40)を発見した。脂質を含むparticleは線状の約26kbp dsDNAで頭部直径約70nmであった。菌体内ではゲノムにintegrateされておらず、おそらくepisomal formで存在するpseudolysogenyで、burst sizeは約13、宿主域は約50%であった。ゲノムおよび性状解析の結果、従来の球形ファージfamily(CorticoviridaeやTectiviridae)には分類できない新規な球形ファージと考えた。(著者抄録)

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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

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    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

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  • 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

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  • 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

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  • 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

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  • 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

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    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

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  • 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

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  • 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

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  • 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

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  • 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

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  • 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

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    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.

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  • 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

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  • 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

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  • 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   2012.2

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  • 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

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  • 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

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    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.

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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

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  • 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

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  • 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

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  • 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

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    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 &lt; .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.

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  • バクテリオファージによるマウス緑膿菌角膜炎の抑制効果

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

    日本眼感染症学会・日本眼炎症学会・日本コンタクトレンズ学会総会プログラム・講演抄録集   48th-45th-54th   51   2011

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  • 黄色ブドウ球菌ファージS13′およびS24‐1ゲノム比較による尾部吸着タンパク質の同定と解析

    内山淳平, 竹村伊代, 佐藤美帆, 加藤伸一郎, 氏原隆子, 松崎茂展, 大畑雅典

    日本ブドウ球菌研究会プログラム・講演要旨集   56th   18   2011

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  • Isolation and characterization of a novel Staphylococcus aureus bacteriophage, phi MR25, and its therapeutic potential

    Hiroshi Hoshiba, Jumpei Uchiyama, Shin-ichiro Kato, Takako Ujihara, Asako Muraoka, Masanori Daibata, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    ARCHIVES OF VIROLOGY   155 ( 4 )   545 - 552   2010.4

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    A novel bacteriophage, phi MR25, was isolated from a lysogenic Staphylococcus aureus strain by mitomycin C induction. Its biological features were analyzed in comparison with phi MR11, which was described previously as a prototype therapeutic phage. phi MR25 is morphologically similar to phi MR11 (morphotype B1 of family Myoviridae) but has a broader host range than phi MR11 on S. aureus strains. phi MR25 can also multiply on S. aureus lysogens of phi MR11. Its DNA is 44,342 bp in size, is predicted to include 70 open reading frames, and does not contain genes related to toxin or drug resistance. The lysogenic module and most of the putative virion protein genes are completely different from those of phi MR11. In spite of their genetic diversity, intraperitoneal administration of phi MR25 rescued mice inoculated with a lethal dose of S. aureus, as was the case for phi MR11. These results suggest that phi MR25 could be another candidate phage to treat S. aureus infection.

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  • 腸球菌ファージφEF24C由来溶菌酵素ORF9の解析

    内山淳平, 林幾江, 竹村伊代, 氏原隆子, 竹内啓晃, 杉浦哲朗, 菅井基之, 大畑雅典, 松崎茂展

    日本細菌学雑誌   65 ( 1 )   180 - 180   2010.2

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  • 新規Serratia marcescensバクテリオファージKSP20,KSP90,およびKSP100の性状解析

    松下憲司, 内山淳平, 氏原隆子, 杉原重喜, 邑岡麻子, 大畑雅典, 松崎茂展

    日本細菌学雑誌   65 ( 1 )   179 - 179   2010.2

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  • マウスにおける治療用候補バクテリオファージの血中動態の比較検討

    内山淳平, 脇口宏, 松崎茂展

    感染症学雑誌   83 ( 5 )   616 - 616   2009.9

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  • Enterococcus faecalisファージφEF24C由来溶菌酵素ORF9の溶菌活性

    竹村伊代, 内山淳平, 竹内啓晃, 杉浦哲朗, 松崎茂展

    感染症学雑誌   83 ( 5 )   612 - 613   2009.9

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  • Blood kinetics of four intraperitoneally administered therapeutic candidate bacteriophages in healthy and neutropenic mice

    UCHIYAMA Jumpei, MAEDA Yoshihiro, TAKEMURA Iyo, CHESS-WILLIAMS Russ, WAKIGUCHI Hiroshi, MATSUZAKI Shigenobu

    Microbiology and immunology   53 ( 5 )   301 - 304   2009.5

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    Due to multiple-drug resistant bacteria, phage therapy is being revisited. Although most animal experiments focus on therapeutic efficacy, the blood clearance kinetics of phages have not been well described. For further development of an efficient therapeutic strategy, information on phage blood kinetics is important. In this study, time-course concentration changes in peripheral blood of healthy and neutropenic mice were measured using four therapeutic phages (φMR11, KPP10, φEF24C, and KEP10). The results showed a two- to three-day rapid phage clearance, which fits a two-compartment model. © 2009 The Societies and Blackwell Publishing Asia Pty Ltd.

    DOI: 10.1111/j.1348-0421.2009.00125.x

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  • Enterococcus faecalisファージφEF24C由来溶菌酵素ORF9の溶菌活性

    竹村 伊代, 内山 淳平, 竹内 啓晃, 杉浦 哲朗, 松崎 茂展

    感染症学雑誌   83 ( 臨増 )   256 - 256   2009.3

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  • マウスにおける治療用候補バクテリオファージの血中動態の比較検討

    内山 淳平, 脇口 宏, 松崎 茂展

    感染症学雑誌   83 ( 臨増 )   260 - 260   2009.3

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  • 緑膿菌の新規溶原性バクテリオファージPAJU2の形態学的・遺伝学的解析

    内山淳平, 松崎茂展

    日本細菌学雑誌   64 ( 1 )   141 - 141   2009.2

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  • Tail-associated structural protein gp61 of Staphylococcus aureus phage phi MR11 has bifunctional lytic activity

    Mohammad Rashel, Jumpei Uchiyama, Iyo Takemura, Hiroshi Hoshiba, Takako Ujihara, Hiroyoshi Takatsuji, Koichi Honke, Shigenobu Matsuzaki

    FEMS MICROBIOLOGY LETTERS   284 ( 1 )   9 - 16   2008.7

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    A tailed bacteriophage, phi MR11 (siphovirus), was selected as a candidate therapeutic phage against Staphylococcus aureus infections. Gene 61, one of the 67 ORFs identified, is located in the morphogenic module. The gene product (gp61) has lytic domains homologous to CHAP (corresponding to an amidase function) at its N-terminus and lysozyme subfamily 2 (LYZ2) at its C-terminus. Each domain of gp61 was purified as a recombinant protein. Both the amidase [amino acids (aa) 1-150] and the lysozyme (aa 401-624) domains but not the linker domain (aa 151-400) caused efficient lysis of S. aureus. Immunoelectron microscopy localized gp61 to the tail tip of the phi MR11 phage. These data strongly suggest that gp61 is a tail-associated lytic factor involved in local cell-wall degradation, allowing the subsequent injection of phi MR11 DNA into the host cytoplasm. Staphylococcus aureus lysogenized with phi MR11 was also lysed by both proteins. Staphylococcus aureus strains on which phi MR11 phage can only produce spots but not plaques were also lysed by each protein, indicating that gp61 may be involved in 'lysis from without'. This is the first report of the presence of a tail-associated virion protein that acts as a lysin, in an S. aureus phage.

    DOI: 10.1111/j.1574-6968.2008.01152.x

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  • In silico and in vivo evaluation of bacteriophage phi EF24C, a candidate for treatment of Enterococcus faecalis infections Reviewed

    Jumpei Uchiyama, Mohammad Rashel, Iyo Takemura, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY   74 ( 13 )   4149 - 4163   2008.7

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    Along with the increasing threat of nosocomial infections by vancomycin-resistant Enterococcus faecalis, bacteriophage (phage) therapy has been expected as an alternative therapy against infectious disease. Although genome information and proof of applicability are prerequisites for a modern therapeutic phage, E. faecalis phage has not been analyzed in terms of these aspects. Previously, we reported a novel virulent phage, phi EF24C, and its biology indicated its therapeutic potential against E. faecalis infection. In this study, the phi EF24C genome was analyzed and the in vivo therapeutic applicability of phi EF24C was also briefly assessed. Its complete genome (142,072 bp) was predicted to have 221 open reading frames (ORFs) and five tRNA genes. In our functional analysis of the ORFs by use of a public database, no proteins undesirable in phage therapy, such as pathogenic and integration-related proteins, were predicted. The noncompetitive directions of replication and transcription and the host-adapted translation of the phage were deduced bioinformatically. Its genomic features indicated that phi EF24C is a member of the SPO1-like phage genus and especially that it has a close relationship to the Listeria phage P100, which is authorized for prophylactic use. Thus, these bioinformatics analyses rationalized the therapeutic eligibility of phi EF24C. Moreover, the in vivo therapeutic potential of phi EF24C, which was effective at a low concentration and was not affected by host sensitivity to the phage, was proven by use of sepsis BALB/c mouse models. Furthermore, no change in mouse lethality was observed under either single or repeated phage exposures. Although further study is required, phi EF24C can be a promising therapeutic phage against E. faecalis infections.

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  • ファージ療法:現状と展望

    松崎茂展, 内山淳平, RASHEL Mohammad, 竹村伊代

    日本細菌学雑誌   63 ( 1 )   56   2008.2

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  • Revival of Bacteriophage ファージ療法 現状と展望

    松崎 茂展, 内山 淳平, Rashel Mohammad, 竹村 伊代

    日本細菌学雑誌   63 ( 1 )   56 - 56   2008.2

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  • MEDICAL TOPICS 第26回 バクテリオファージを用いる難治性感染症治療

    松崎茂展, 内山淳平, RASHEL Mohammad, 竹村伊代

    Lung Perspect   16 ( 1 )   98 - 103   2008.1

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  • MEDICAL TOPICS バクテリオファージを用いる難治性感染症治療

    松崎 茂展, 内山 淳平, Rashel Mohammad, 竹村 伊代

    THE LUNG-perspectives   16 ( 1 )   98 - 103   2008.1

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    [仮説]ファージ療法は、欧米諸国では抗生物質の実用化に伴い一旦放棄されたが、旧ソ連諸国ポーランドでは現在に至るまで使用されている。ファージ療法の導入により、現在の化学療法の弱点(耐性菌、菌交代症の問題など)を補足することは可能であろうか。[新知見の要点]欧米の多くの研究者によってよく計画された動物実験により、ファージ療法(活性ファージおよびライシンの使用)の有効性・安全性を示すデータが蓄積されつつある。[将来の可能性、問題点]今後、各菌種に対応可能な実用的ファージバンクの構築が必要である。(著者抄録)

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  • Isolation and characterization of a novel Enterococcus faecalis bacteriophage phi EF24C as a therapeutic candidate Reviewed

    Jumpei Uchiyama, Mohammad Rashel, Yoshihiro Maeda, Iyo Takemura, Shigeyoshi Sugihara, Kazue Akechi, Asako Muraoka, Hiroshi Wakiguchi, Shigenobu Matsuzaki

    FEMS MICROBIOLOGY LETTERS   278 ( 2 )   200 - 206   2008.1

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    Vancomycin-resistant Enterococcus faecalis (VRE) has become a significant threat in nosocomial settings. Bacteriophage (phage) therapy is frequently proposed as a potential alternative therapy for infections caused by this bacterium. To search for candidate therapeutic phages against Enterococcus faecalis infections, 30 Enterococcus faecalis phages were isolated from the environment. One of these, virulent phage phi EF24C, which has a broad host range, was selected for analysis. The plaque-forming ability of phi EF24C was virtually unaffected by differences in the clinical host strains. Furthermore, the phage had a shorter latent period and a larger burst size than ordinary tailed phages, indicating that phi EF24C has effective lytic activity against many Enterococcus faecalis strains, including VRE. Morphological and genomic analyses revealed that phi EF24C is a large myovirus (classified as family Myoviridae morphotype A1) with a linear double-stranded DNA genome of c. 143 kbp. Analyses of the N-terminal amino acid sequences of the virion proteins, together with the morphology and the genome size, speculated that phi EF24C is closely related to other myoviruses of Gram-positive bacteria that have been used experimentally or practically for therapy or prophylaxis. Considering these results, phi EF24C may be a potential candidate therapeutic phage against Enterococcus faecalis infections.

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  • A novel site-specific recombination system derived from bacteriophage $ϕ$MR11 Reviewed

    Rashel, Mohammad, Uchiyama, Jumpei, Ujihara, Takako, Takemura, Iyo, Hoshiba, Hiroshi, Matsuzaki, Shigenobu

    Biochemical and biophysical research communications   368 ( 2 )   192 - 198   2008

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    We report identification of a novel site-specific DNA recombination system that functions in both it? vivo and ill Vitro, derived from lysogenic Staphylococcus aureus phage phi MR11. In silico analysis of the phi MR11 genome indicated orf1 as a putative integrase gene. Phage and bacterial attachment sites (attP and attB, respectively) and attachment junctions were determined and their nucleotide sequences decoded. Sequences of attP and attB were mostly different to each other except for a two bp common core that was the crossover point. We found several inverted repeats adjacent to the core sequence of attP as potential protein binding sites. The precise and efficient integration properties of phi MR11 integrase were shown on attP and attB in Escherichia coli and the minimum size of attP was found to be 34 bp. In in vitro assays using crude or purified integrase, only buffer and substrate DNAs were required for the recombination reaction, indicating that other bacterially encoded factors are not essential for activity. (C) 2008 Elsevier Inc. All rights reserved.

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  • T-even-related bacteriophages as candidates for treatment of Escherichia coli urinary tract infections Reviewed

    Nishikawa, H, Yasuda, M, Uchiyama, J, Rashel, M, Maeda, Y, Takemura, I, Sugihara, S, Ujihara, T, Shimizu, Y, Shuin, T, others

    Archives of virology   153 ( 3 )   507 - 515   2008

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    Multidrug-resistant uropathogenic Escherichia coli (UPEC) is increasing gradually on a worldwide scale. We therefore examined the possibility of bacteriophage (phage) therapy for urinary tract infections (UTIs) caused by the UPEC strains as an alternative to chemotherapy. In addition to the well-known T4 phage, KEP10, which was newly isolated, was used as a therapeutic phage candidate. KEP10 showed a broader bacteriolytic spectrum (67%) for UPEC strains than T4 (14%). Morphological and genetic analyses showed that KEP10 resembles phage T4. Phages T4 and KEP10 injected into the peritoneal cavity of mice were distributed immediately to all organs examined and maintained a high titer for at least 24 h. They were stable in the urine of both mice and humans for 24 h at 37 degrees C. Administration of these phages into the peritoneal cavity caused a marked decrease in the mortality of mice inoculated transurethrally with a UPEC strain, whereas most of the control mice died within a few days of bacterial infection. Inoculation with phage alone produced no adverse effects attributable to the phage per se. The present study experimentally demonstrated the therapeutic potential of phage for E. coli-induced UTIs, and T-even-related phages may be suitable candidates with which to treat them.

    DOI: 10.1007/s00705-007-0031-4

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  • 腸球菌感染症に対する治療用バクテリオファージφEF24Cの性状とゲノム解析

    内山淳平, 松崎茂展, MOHAMMAD Rashel, 前田良浩, 竹村伊代, 邑岡麻子, 今井賞介

    感染症学雑誌   81 ( 5 )   607   2007.9

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  • ファージφMR11のtail関連成分GP61の機能的解析 φMR11由来staphylolytic物質の他の可能性(Functional analysis of the tail-associated component GP61 of phage φMRl11: another possible φMR11-derived staphylolytic agent)

    Mohammad Rashel, 松崎 茂展, 内山 淳平, 前田 良浩, 竹村 伊代, 今井 章介

    感染症学雑誌   81 ( 5 )   646 - 646   2007.9

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  • 薬剤耐性と対策 GPC 腸球菌感染症に対する治療用バクテリオファージφEF24Cの性状とゲノム解析

    内山 淳平, 松崎 茂展, Mohammad Rashel, 前田 良浩, 竹村 伊代, 邑岡 麻子, 今井 賞介

    感染症学雑誌   81 ( 5 )   607 - 607   2007.9

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  • Phage φMR11のtail関連成分GP61の機能的解析 φMR11由来staphylolytic物質の他の可能性(Functional analyis of the tail-associated component GP61 of phage φMR11: another possible φMR11-derived staphylolytic agent)

    Mohammad Rashel, 松崎 茂展, 内山 淳平, 前田 良浩, 竹村 伊代, 今井 章介

    感染症学雑誌   81 ( 臨増 )   229 - 229   2007.3

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  • 薬剤耐性と対策 GPC 腸球菌感染症に対する治療用バクテリオファージφEF24Cの性状とゲノム解析

    内山 淳平, 松崎 茂展, Mohammad Rashel, 前田 良浩, 竹村 伊代, 邑岡 麻子, 今井 章介

    感染症学雑誌   81 ( 臨増 )   165 - 165   2007.3

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  • 腸球菌感染症に対する治療用バクテリオファージφEF24Cのゲノム解析

    内山 淳平, 松崎 茂展, モハンマド ラシェル, 前田 良浩, 竹村 伊代, 黒田 正幸, 邑岡 麻子, 今井 章介

    日本細菌学雑誌   62 ( 1 )   144 - 144   2007.2

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  • In vivo and in vitro site-specific DNA recombination by phage φMR11-derived machinery

    モハンマド ラッシェル, 松崎 茂展, 黒田 正幸, 藤原 隆子, 内山 淳平, 前田 良浩, 竹村 伊代, 今井 章介

    日本細菌学雑誌   62 ( 1 )   141 - 141   2007.2

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  • ファージφMR11-由来機構によるIn vivoおよびin vitro部位特異的DNA組換え(In vivo and in vitro site-specific DNA recombination by phage φ MR11-derived machinery)

    ラッシェル・モハンマド, 松崎 茂展, 黒田 正幸, 藤原 隆子, 内山 淳平, 前田 良浩, 竹村 伊代, 今井 章介

    日本細菌学雑誌   62 ( 1 )   141 - 141   2007.2

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  • バクテリオファージ療法

    内山淳平, 松崎茂展, 今井章介

    医学のあゆみ   219 ( 6 )   483 - 484   2006.11

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  • 細菌学・ウイルス学 バクテリオファージ療法

    内山 淳平, 松崎 茂展, 今井 章介

    医学のあゆみ   219 ( 6 )   483 - 484   2006.11

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  • 腸球菌感染症に対するファージ療法の検討

    明智和愛, 松崎茂展, 内山淳平, RASHEL M, 桜井慎吾, 氏原隆子, 邑岡麻子, 杉原重喜, 杉浦哲朗, 今井章介

    こうち   36 ( 1 )   12 - 12   2006.10

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  • バクテリオファージ療法:その現状と将来展望

    今井章介, 松崎茂展, RASHEL Mohammad, 内山淳平

    感染症学雑誌   80 ( 5 )   549 - 550   2006.9

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  • ワクチン,抗菌薬以外のアプローチ

    松崎茂展, RASHEL Mohammad, 内山淳平, 今井章介

    感染症学雑誌   80 ( 5 )   556 - 557   2006.9

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  • 小児科外来診療における耐性菌の現状と対策 ワクチン、抗菌薬以外のアプローチ

    松崎 茂展, Rashel Mohammad, 内山 淳平, 今井 章介

    感染症学雑誌   80 ( 5 )   556 - 557   2006.9

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  • 小児科外来診療における耐性菌の現状と対策 ワクチン,抗菌薬以外のアプローチ

    松崎 茂展, Rashel Mohammad, 内山 淳平, 今井 章介

    感染症学雑誌   80 ( 臨増 )   103 - 103   2006.3

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  • バクテリオファージ療法 その現状と将来展望

    今井 章介, 松崎 茂展, Rashel Mohammad, 内山 淳平

    感染症学雑誌   80 ( 臨増 )   92 - 92   2006.3

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  • 腸球菌感染症に対するファージ療法の検討:(2) 実験的腸球菌感染マウスに対するファージ投与効果

    明智和愛, 内山淳平, 松崎茂展, モハンマド ラッシェル, 干場浩, 桜井慎吾, 前田良浩, 氏原隆子, 邑岡麻子, 今井章介

    日本細菌学雑誌   61 ( 1 )   112 - 112   2006.2

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  • 腸球菌感染症に対するファージ療法の検討:(1) 新規腸球菌ファージの分離と性状解析

    内山淳平, 明智和愛, 松崎茂展, モハンマド ラッシェル, 黒田正幸, 小谷典弘, 上原良雄, 杉原重喜, 杉浦哲朗, 今井章介

    日本細菌学雑誌   61 ( 1 )   111 - 111   2006.2

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  • Bacteriophage therapy : a revitalized therapy against bacterial infectious diseases

    MATSUZAKI Shigenobu, RASHEL Mohammad, UCHIYAMA Jumpei, SAKURAI Shingo, UJIHARA Takako, KURODA Masayuki, IKEUCHI Masahiko, TANI Toshikazu, FUJIEDA Mikiya, WAKIGUCHI Hiroshi, IMAI SHOSUKE

    11 ( 5 )   211 - 219   2005.10

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    Bacteriophage (phage) therapy involves using phages or their products as bioagents for the treatment or prophylaxis of bacterial infectious diseases. Much evidence in support of the effectiveness of phage therapy against bacterial infectious diseases has accumulated since 1980 from animal model studies conducted in Western countries. Reports indicate that appropriate administration of living phages can be used to treat lethal infectious diseases caused by gram-negative bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Vibrio vulnificus, and Salmonella spp., and gram-positive bacteria, such as Enterococcus faecium and Staphylococcus aureus. The phage display system and genetically modified nonreplicating phages are also effective for treatment of Helicobacter pylori and P. aeruginosa, respectively. In addition to phage particles per se, purified phage-encoded peptidoglycan hydrolase (lysin) is also reported to be effective for the treatment of bacterial infectious diseases caused by gram-positive bacteria such as Streptococcus pyogenes, S. pneumoniae, Bacillus anthracis, and group B streptococci. All phage lysins that have been studied to date exhibit immediate and strong bacteriolytic activity when applied exogenously. Furthermore, phage-coded inhibitors of peptidoglycan synthesis (protein antibiotics), search methods for novel antibacterial agents using phage genome informatics, and vaccines utilizing phages or their products are being developed. Phage therapy will compensate for unavoidable complications of chemotherapy such as the appearance of multidrug resistance or substituted microbism. © Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases 2005.

    DOI: 10.1007/s10156-005-0408-9

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  • ヒト細胞で機能する黄色ブドウ球菌ファージφMR11由来部位特異的組換えシステム

    氏原隆子, 松崎茂展, 黒田正幸, RASHEL M, 明智和愛, 内山淳平, 桜井慎吾, 邑岡麻子, 今井章介

    日本細菌学雑誌   60 ( 1 )   63 - 63   2005.2

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Industrial property rights

  • 酸素ナノバブルを含む医薬

    永根 大幹, 内山 淳平, 山下 匡, 杉本 義孝

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    Application no:特願2021-157759  Date applied:2021.9.28

    Announcement no:特開2022-056407  Date announced:2022.4.8

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  • アレルゲン不活化剤の評価方法及びアレルゲン不活化剤の評価キット

    阪口 雅弘, 内山 淳平, 吉田 愛美, 水上 圭二郎, 蔵田 圭吾

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    Application no:JP2021016529  Date applied:2021.4.23

    Patent/Registration no:特許第7126665号  Date registered:2022.8.19 

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  • 乳酸菌含有組成物

    阪口 雅弘, 内山 淳平, 福山 朋季

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    Application no:特願2020-038610  Date applied:2020.3.6

    Announcement no:特開2021-136951  Date announced:2021.9.16

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  • バクテリオファージ由来エンドライシン

    内山 淳平, 金木 真央

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    Applicant:日本全薬工業株式会社

    Application no:特願2020-006483  Date applied:2020.1.20

    Announcement no:特開2021-112152  Date announced:2021.8.5

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  • 新規バクテリオファージおよび細菌性眼内炎治療剤

    福田 憲, 松崎 茂展, 福島 敦樹, 大畑 雅典, 内山 淳平

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    Applicant:国立大学法人高知大学

    Application no:JP2019009112  Date applied:2019.3.7

    Publication no:WO2019-176729  Date published:2019919

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  • 細菌選択培地

    内山 淳平, 村上 裕信

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    Application no:特願2018-114145  Date applied:2018.6.15

    Announcement no:特開2019-216609  Date announced:2019.12.26

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  • B群連鎖球菌選択的増菌用培地

    内山 淳平, 阪口 雅弘, 花木 秀明, 松井 秀仁

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    Applicant:学校法人麻布獣医学園, 学校法人北里研究所

    Application no:特願2016-247315  Date applied:2016.12.21

    Announcement no:特開2018-099080  Date announced:2018.6.28

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  • 黄色ブドウ球菌を検出する方法、黄色ブドウ球菌測定用メンブレン、及び黄色ブドウ球菌測定用キット

    内山 淳平, 内山 伊代, 花木 秀明, 松井 秀仁, 大畑 雅典, 松崎 茂展

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    Applicant:学校法人麻布獣医学園, 学校法人北里研究所

    Application no:特願2014-046922  Date applied:2014.3.10

    Announcement no:特開2015-169632  Date announced:2015.9.28

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  • 黄色ブドウ球細菌に結合するタンパク質及びそのタンパク質を利用した黄色ブドウ球細菌の測定方法。

    大畑 雅典, 松崎 茂展, 内山 淳平, 竹村 伊代

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    Applicant:学校法人麻布獣医学園

    Application no:特願2012-160613  Date applied:2012.7.19

    Announcement no:特開2013-066456  Date announced:2013.4.18

    Patent/Registration no:特許第5925075号  Date registered:2016.4.28  Date issued:2016.4.28

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  • 黄色ブドウ球細菌に結合するタンパク質及びそのタンパク質を利用した黄色ブドウ球細菌の測定方法。

    大畑 雅典, 松崎 茂展, 内山 淳平, 竹村 伊代

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    Applicant:国立大学法人高知大学

    Application no:特願2012-160613  Date applied:2012.7.19

    Announcement no:特開2013-066456  Date announced:2013.4.18

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Awards

  • 創立四半世紀記念研究成果発表奨励賞

    2022.3   日本臨床微生物学会  

    内山淳平

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  • 第68回日本細菌学会中国・四国支部総会 若手研究者奨励賞受賞

    2015.9  

    内山淳平

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  • 高知大学研究顕彰制度「若手教員研究優秀賞」

    2014.3  

    内山淳平

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  • 高知信用金庫・安心友の会 学術賞

    2014.2  

    内山淳平

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  • 大学院生奨励賞

    2009.3   高知大学  

    内山淳平

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  • エンデバー賞

    2008.1   オーストラリア政府  

    内山淳平

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  • Dean’s Commendation, Faculty of Science

    2004.12   University of the Sunshine Coast, Australia  

    内山淳平

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Research Projects

  • ファージ吸着分子結合マイクロ粒子を利用した抗C. difficile治療法の創出

    Grant number:22K08580  2022.04 - 2025.03

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    内山 淳平

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    Grant amount:\4160000 ( Direct expense: \3200000 、 Indirect expense:\960000 )

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  • KHP30様ファージのピロリ菌認識機構の解明とピロリ菌の迅速検査・除菌への応用

    Grant number:22K08593  2022.04 - 2025.03

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    松崎 茂展, 内山 淳平, 岩本 昌大

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    Grant amount:\4160000 ( Direct expense: \3200000 、 Indirect expense:\960000 )

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  • Development of a novel detection method for whole bacterial cells using bacteriophages as recognition elements

    Grant number:21K05113  2021.04 - 2024.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    渡辺 茂, 内山 淳平, 仁子 陽輔

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    Grant amount:\4160000 ( Direct expense: \3200000 、 Indirect expense:\960000 )

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  • Study for a molecular machinery of the host recognition by Clostridioides difficile phage to develop the advanced therapy

    Grant number:19K07560  2019.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    阪口 義彦, 加藤 はる, 大宮 直木, 内山 淳平, 後藤 和義, 妹尾 充敏, 林 俊治

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    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    クロストリディオイデス(クロストリジウム)・ディフィシル感染症(CDI)は、抗菌薬関連下痢症・腸炎の1つであり、抗菌薬の使用等により正常な腸内微生物叢が乱れ、ディフィシル菌が過剰に増殖することにより発症する。CDI治療では、バンコマイシンやメトロニダゾールなどの抗菌薬が有効であるが、抗菌薬は選択性に乏しく結局は有益な腸内微生物叢を撹乱させるため、再びCDIが引き起こされることがある。従って、腸内微生物叢を撹乱せず、原因となるディフィシル菌のみを特異的に殺菌するCDIの新規治療法の確立が喫緊である。本研究課題では、新規治療法の候補の1つとして、バクテリオファージ(ファージ)に着目した。ファージは、宿主である細菌に対して特異的に結合し、自己の溶菌酵素により殺菌する活性を示す。このようなファージの特性を有効に利用したCDIの新規治療法の確立を目指し研究開発を行っている。そこで、まず、溶菌性ファージの単離を試みたが、分離株からは困難であったことから、次に、ファージが有する溶菌酵素について解析を行った。ディフィシル菌ファージのゲノム情報を基に、ゲノム上の個々の遺伝子について他のファージの溶菌酵素との相同性解析を行ったところ、2つのファージ由来溶菌酵素と予想される遺伝子(cdph33およびcdph34)を特定した。そこで、これらの遺伝子を基にHis融合タンパク質(CDph33およびCDph34)として、大腸菌で発現・精製した。精製した2つのタンパク質について、ディフィシル菌に対する溶菌活性を測定したところ、CDph34は活性を示したが、CDph33は高濃度下でも活性が認められなかった。CDph33とCDph34を組み合わせた場合では、CDph34の溶菌活性と比較してほぼ同等の活性であったが、両タンパク質を組み合わせた方が時間依存的に高い活性を示した。現在、詳細な解析を進めている。

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  • Development of the new multidrug-resistant tuberculosis control method to use bacteriophage lysin

    Grant number:19K08953  2019.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    松崎 茂展, 山本 哲也, 内山 淳平, 渡辺 茂, 福田 憲, 北村 直也

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    本件研究では、ファージY10が保有する溶菌酵素(ライシン)を利用する結核菌制御法の開発を目指している。低病原性迅速発育抗酸菌であるスメグマ菌は、ファージY10に対して結核菌と同様の感受性を示すことが知られている。
    そのため、まずスメグマ菌に対するライシンの溶菌測定条件の検討を行なった。抗酸菌は、コードファークター産生の故に、液体培地では紐状構造となるため、吸光度測定を使用するファージやファージライシンのインビトロ活性の計測が極めて困難である。そのため、培地中に結核菌培養で使用されるTween20をLB培地に添加すると、菌は均一になるものの、ファージの増殖活性が著しく抑制されることが分かった。そのため、ライシンに対しても抑制的に働く可能性があるため、他の界面活性剤の有用性をスクリーニングすることにした。まず、9種類の界面活性剤について、0.5%濃度の添加により、ファージY10の増殖に影響があるか否かを検討した。その結果、CHAPS,n-Octyl-β-D-glucoside, Sodium cholate, MEGA-8,n-octyl-β-D-maltosideの4種においては、ファージの活性は大きく低下することはなかった。次に、これら4種の界面活性剤を各種濃度で添加したLBにおける、菌の増殖及び菌拡散状況について検討した。その結果、これら4種の界面活性剤とも、0.01~0.025%の濃度で菌増殖が正常に増殖できることが分かった。また、Tween20ほど強力ではないものの、対照と比較して菌を均一化する拡散効果が認められれた。一方、物理的方法による菌の均一化についても検討した。菌培養液を、短時間超音波処理すると、菌の生存率を落とすことなく均一化できることが分かった。

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  • Development of novel bacterial detection methods using bacteriophages as recognition elements

    Grant number:18K05174  2018.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    渡辺 茂, 松崎 茂展, 仁子 陽輔, 内山 淳平

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    微生物検査は,感染症や食中毒の予防・衛生管理の面から重要である.しかし,培養に基づく従来法では検査に2~10日を要し,病原菌の特定に至っても手遅れになる場合が多く,検査の迅速化・簡易化が強く求められている.近年,PCRや抗原抗体反応を利用した迅速検査技術が開発されているが,遺伝子解析には煩雑な操作や高額な機器が必要である.また,細菌の中には,宿主免疫を回避するような防御機能を備えているものもあり,遺伝子解析や免疫反応に依らない新奇な検出原理が求められている.
    バクテリオファージ(以下,ファージと省略する)は,細菌特異的に感染するウイルスの総称であり,標的細菌を特異的に識別する機能を備えている.その機能を自在に活用できるようになれば,遺伝子や免疫検出法に代わる迅速かつ簡便な細菌検出法への応用が期待できる.本年度は,ファージと金ナノ粒子を組み合わせた比色法に基づく細菌検出技術について検討した.
    ファ―ジは,その表面のカルボキシル基を活性エステルへと変換した後,アミノエタンチオールと反応させることによってチオール化ファージへ変換した.次に細菌を含む試料と混合した後,遠心分離によって余剰のファージを取除き,金ナノ粒子の水溶液を添加した.黄色ブドウ球菌(S.aureus)を宿主とするファージを利用した場合,試料にS.aureusが含まれている場合,金ナノ粒子溶液の色は赤色から青色へと色調が変化する様子が観察された.一方,大腸菌属のE.coliや同じブドウ球菌属のS.Pseudintermediusを含む試料では色調変化は全く観察されなかった.本手法は,特別な分析機器を利用することなく,属のみならず種レベルで細菌を高選択的に検出することが可能であり,検出限界は数百cfu/mlに達することがわかった.また,利用するファージを変えることによって細菌選択性を自在に制御できる可能性がある.

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  • Structure analysis of Staphylococcus bacteriophage S13'

    Grant number:18K06154  2018.04 - 2021.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Miyazaki Naoyuki

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    We analyzed the infection mechanisms of Staphylococcus bacteriophages S13’ and S6 by cryo-electron microscopy for practical and effective application of phage therapy for the Staphylococcus aureus infectious diseases. The S13’ is a small phage in the Podoviridae family with a head diameter of about 50 nm and a non-contractile tail length of about 40 nm, while the S6 is a large Myoviridae phage with a diameter of about 140 nm and a contractile tail length of about 250 nm. We succeeded to build atomic models for almost all of the proteins that make up the phage particle. On the other hand, we determined the tail structures of S6 before and after contraction, and then we reveals the contractile mechanism of the tail.

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  • Examination of the persistent infection mechanism of Helicobacter pylori by phage lysogenization

    Grant number:18K07127  2018.04 - 2021.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Uchiyama Jumpei

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    Grant amount:\4550000 ( Direct expense: \3500000 、 Indirect expense:\1050000 )

    Helicobacter pylori is a pathogenic bacterium involved in diseases, for example, chronic gastritis, gastric ulcer, and gastric cancer. Since phages often have useful genes for host bacteria, phage lysogenization to bacteria can confer the bacteria the traits such as survival strategy in the environment and pathogenicity. In recent years, lysogenic phage infecting H. pylori has been discovered. Since many defective phages have been often found In the H. pylori genomes, the traits are expected to be conferred to H. pylori by phage infections. However, the traits of H. pylori by phage lysogenization are hardly understood. In this study, lysogenization of H. pylori phages was clarified to improve the bacterial mobility.

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  • Pathogenicity analysis of bovine leukemia virus

    Grant number:17K15385  2017.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)  Grant-in-Aid for Young Scientists (B)

    Murakami Hironobu, Sato Reiichiro, Uchiyama Jumpei, Maeda Yosuke, Kato Hajime

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    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    To research whether a genetic variation affects viral properties, phylogenetic analysis was performed using whole BLV genome and revealed to be classified three group in BLV wild-type strains. In addition, the groups were associated with virus productivity. The one of three group strains classified in this study showed high virus productivity and had an advantage for transmissibility in field. Moreover, pathogenesis was different among wild-type strains, but some strains harbored deletion mutation and showing low viral production had high pathogenesis. Taken together, transmissibility and pathogenesis would be independently decided by different genetic factors.

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  • Study of the molecular machinery of host recognition by the Clostridium botulinum type C and D neurotoxin-converting phages

    Grant number:16K08785  2016.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Sakaguchi Yoshihiko

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    Grant amount:\4810000 ( Direct expense: \3700000 、 Indirect expense:\1110000 )

    Our ongoing study revealed that botulinum neurotoxin genes are present in the bacteriophage (phage) genomes and that, consequently, phages participate in neurotoxin-gene transfer to Clostridium botulinum types C and D. If so, a strategy mediated by phages to inhibit neurotoxin-gene transfer to the hosts would be of great help to block botulism outbreaks. In this study, we aimed to identify the infectious mechanism of C. botulinum type C and D phages. We performed genomics analyses of type C and D phages, which was useful to specify the phage adsorption rate that is required for adherence to the host surface. We found that phages carry several interesting genes adapted to our purpose, which are likely to encode the proteins that compose the phage tail. These novel findings will be very useful to identitify the machinery of the phage host interaction in the process of infection-mediated neurotoxin-gene transfer.

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  • Basic research to establish Bacteriophage treatment for chronic otitis media as a novel treatment strategies using the activity of bacteriolysis

    Grant number:15K10756  2015.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Komori Masahiro

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    Grant amount:\4810000 ( Direct expense: \3700000 、 Indirect expense:\1110000 )

    Bacteriophage is a virus which infects to bacterium and breaks them. Since having specificity to host bacterium, bacteriophage never infects to the indigenous bacterium and human cells. Thus, when host bacterium disappears, the bacteriophage becomes extinct. In addition, it does not have tolerance to antibiotics and also breaks the biofilms of bacterium.
    This study aimed to confirm the efficacy and safety on the treatments for chronic otitis media. Staphylococcus aureus and/or bacteriophage were injected into the middle ear cavity. Although their concentration to work was much higher than expected, the inflammation disappeared by the bacteriophage treatment. No mice with bad condition, death and doubted inner ear damages were found. The study indicated the bacteriophage is effective for the treatments to the otitis media, however, further studies will be required for the toxicity.

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  • Analysis of MRSA evolution mechanism by generalized transduction

    Grant number:15K19095  2015.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)  Grant-in-Aid for Young Scientists (B)

    UCHIYAMA JUMPEI, MATSUZAKI Shigenobu, KUROKAWA Kenji, MIYAZAKI Naoyuki, MURATA Kazuyoshi, UCHIYAMA Iyo

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    Grant amount:\4030000 ( Direct expense: \3100000 、 Indirect expense:\930000 )

    In this study, we analyzed the horizontal gene transfer in the environment, using the model of phage transduction of phage S6 in MRSA. First, analyzing phylogeny of phage based on the whole genome, phage S6 was related to the other phage infecting Gram-positive bacteria and that infecting Gram-negative bacteria, which suggested that there may be specific phage group with transducing ability. Second, phage S6 has ability to adsorb the Staphylococcus aureus cells via beta-glycosilation of wall teichoic acids on the cell wall. Third, phage S6 was considered to package the host DNA, transfer it to the new recipient cells, and the gene acquisition occurred by homologous recombination.

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  • Development of novel treatment by using bacteriophage for ocular diseases induced by bacterial infection

    Grant number:26462692  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    FUKUDA Ken

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    Grant amount:\4940000 ( Direct expense: \3800000 、 Indirect expense:\1140000 )

    We examined the therapeutic effects of bacteriophage eye drop on methicillin-resistant Staphylococcus aureus keratitis in rabbit. The clinical corneal score was significantly lower in rabbits treated by bacteriophage eye drop than in those treated by anti-biotic eye drop or control. The bacterial load in cornea among the vehicle-treated rabbit, anti-biotic-treated rabbit, and bacteriophage-treated rabbit were not significantly changed.
    We then examined the bacterial endophthalmitis in rabbit and mouse. The injection Pseudomonas aeruginosa or Enterococcus faecalis into vitreous cavity in mice stably induced severe endophthalmitis including massive infiltration of neutrophils and destruction of tissue. Injection of phage into vitreous cavity in mice with endophthalmitis significantly suppressed clinical score and viable bacteria in the eyeball.

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  • Elucidation of a relationship of bacteriophage to pathogenicity of Helicobacter pylori and development of eradication system to H. pylori using bacteriophage.

    Grant number:26461504  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    MATSUZAKI Shigenobu

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    Grant amount:\4940000 ( Direct expense: \3800000 、 Indirect expense:\1140000 )

    In this study, we aimed at collection of information about a bacteriophage-exclusion system (restriction-modification system) in Helicobacter pylori, to develop an effective sterilization method of drug-resistant H. pylori from the stomach using bacteriophages KHP30 and KHP40. The results indicated that H. pylori had multiple functional restriction-modification systems, suggesting strongly that the bacteriophages must be adapted to all H. pylori-possessed restriction-modification systems for effective sterilization.

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  • Analysis of tail adsorbed protein of Clostridium botulinum types C and D neurotoxin-converting phage and its host bacterial receptors

    Grant number:25460541  2013.04 - 2016.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Sakaguchi Yoshihiko, MATSUZAKI Shigenobu, UCHIYAMA Jumpei

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    Grant amount:\5200000 ( Direct expense: \4000000 、 Indirect expense:\1200000 )

    Because Clostridium botulinum types C and D produce botulinum neurotoxin, causing serious symptom botulism, they are recognized as severe pathogen you should prevent spreading in food and animal industries. The botulinum neurotoxin genes are coded on the prophage genomes, and so inhibition of lateral gene transfer mediated by bacteriophage (phage) can be one of the preventions for the disease. Thus, we investigated the adsorption mechanism of C. botulinum type C neurotoxin-converting phage, phage c-st on the genetic point. Based on the comparative genomics and identification of phage virion genes, we got at a speculation that several genes associated with phage adsorption. We are in advanced studies of the findings.

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  • ポイント・オブ・ケア診断を目指したバクテリオファージ尾部吸着分子を利用した簡易迅速細菌検出技術の開発

    2013.04 - 2014.03

    黒住医学研究振興財団  第21回研究助成金  第21回研究助成金

    内山淳平

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    Authorship:Principal investigator 

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  • Examination of anti-viral activity of novel phage-derived nucleosides

    Grant number:24791025  2012.04 - 2015.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)  Grant-in-Aid for Young Scientists (B)

    UCHIYAMA Jumpei, GAMOH Keiji

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    Nucleic acid analogs, which the structure of the nucleoside is based, is used in chemotherapy against viral infections. Some of the bacteriophages (phages) have unique genomic DNAs. In this study, we searched novel nucleic acids from phage genomic DNA, which can become drug discovery platform for anti-viral drugs.
    The phage genomic DNAs were analyzed using LC-MS. As a result, most of the phages were revealed to have normal genomic DNAs. On the other hand, Staphylococcus aureus phage S6 DNA was revealed to contain uracil in place of thymine as a nucleic acid base. Unfortunately, it did not lead to the discovery of novel nucleic acids that can be drug discovery platform for anti-viral drugs in the present study.

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  • Development of the new Helicobacter pylori eradication method for using a phage lytic enzyme

    Grant number:23591478  2011 - 2013

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    MATSUZAKI Shigenobu, TAKEUCHI Hiroaki, UCHIYAMA Jumpei, DAIBATA Masanori

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    Authorship:Collaborating Investigator(s) (not designated on Grant-in-Aid)  Grant type:Competitive

    Grant amount:\5330000 ( Direct expense: \4100000 、 Indirect expense:\1230000 )

    To develop a novel eradication method of Helicobater pylori using bacteriolytic activity of a bacteriophage (phage) KHP30, the morphological, physical, and genetic aspects and the life-cycle of KHP30 have been studied. The host-range of KHP30 was about 65%, suggesting that it is suitable for application use as antibacterial agent. The complete DNA nucleotide sequence was determined and the open reading frames (ORFs) were annotated. The genome consisted of 26 kbp including 30 ORFs. By homology search of the genome, the ORF for the holin protein, one component of general lytic system, was found, but the ORF for the lysin, the counterpart of the system was not found. Therefore, H. pylori phage KHP30 was supposed to have a unique bacteriolytic system different from those of other phages.

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  • Development of a new rapid bacterial identification using bacteriophage tail ligand molecule

    Grant number:22890129  2010 - 2011

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Research Activity Start-up  Grant-in-Aid for Research Activity Start-up

    UCHIYAMA Jumpei, DAIBATA Masanori, MATSUZAKI Shigenobu, UCHIYAMA Iyo

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3016000 ( Direct expense: \2320000 、 Indirect expense:\696000 )

    Infection control is extremely important to prevent infection spread. Rapid bacterial examination in infection control enables early treatment of patients, reduction of medical costs and prevention of infection spread. Thus, the development of more rapid bacterial detection systems is urgently required. The principal Investigator focused on the specific bacteriophage(phage ; bacterial virus) structural proteins contributed to the bacteria-species-specific binding ability, and examined the applicability of the phage molecule to bacterial detection system. As a result, the phage tail molecule was revealed to be potentially used for the rapid bacteria detection system.

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Class subject in charge

  • Combat against Infectious Diseases (2022academic year) Second semester  - 月1~2

  • Microbiology (2022academic year) special  - その他

  • Practice in Microbiology (2022academic year) special  - その他