Updated on 2022/07/15

写真a

 
KAITO Chikara
 
Organization
Faculty of Medicine, Dentistry and Pharmaceutical Sciences Professor
Position
Professor
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Degree

  • Ph.D. ( The University of Tokyo )

Research Interests

  • 感染

  • モデル動物

  • 自然免疫

  • 毒素

  • 黄色ブドウ球菌

  • 病原性

  • カイコ

  • 感染症

  • 細菌学

  • 微生物学

  • 運動

  • RNA

  • 免疫

  • mobile genetic element

  • 薬剤耐性

  • 昆虫

  • 病原性微生物

  • Staphylococcus aureus

  • Colony Spreading

  • Silkworm model

Research Areas

  • Life Science / Bacteriology

Education

  • The University of Tokyo   理学部   生物学科 動物学専攻 卒業

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    Country: Japan

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  • The University of Tokyo   大学院薬学系研究科   博士課程 修了

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    Country: Japan

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Research History

  • Okayama University   Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences   Professor

    2019.1

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  • The University of Tokyo   Graduate School of Pharmaceutical Sciences   Associate Professor

    2010.7 - 2018.12

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  • The University of Tokyo   Graduate School of Pharmaceutical Sciences   Lecturer

    2008.3 - 2010.6

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  • The University of Tokyo   Graduate School of Pharmaceutical Sciences   Assistant Professor

    2007.4 - 2008.2

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  • The University of Tokyo   Graduate School of Pharmaceutical Sciences   Research Assistant

    2005.7 - 2007.3

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Professional Memberships

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Committee Memberships

  • 日本細菌学会   理事  

    2021.1   

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    Committee type:Academic society

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  • Microbiology and Immunology   Deputy Editors-in-Chief  

    2021.1   

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    Committee type:Academic society

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  • 日本医療研究開発機構   AMED委員  

    2017.4   

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    Committee type:Government

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  • 日本学術振興会   科学研究費 審査委員  

    2017   

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    Committee type:Government

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  • 日本ブドウ球菌研究会   運営委員  

    2016.6   

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    Committee type:Academic society

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  • PLOS ONE   Editorial Board  

    2016.6   

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    Committee type:Academic society

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Papers

  • Knockout of mlaA increases Escherichia coli virulence in a silkworm infection model Reviewed

    PLOS ONE   17 ( 7 )   e0270166   2022.7

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    DOI: 10.1371/journal.pone.0270166

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  • The absence of osmoregulated periplasmic glucan confers antimicrobial resistance and increases virulence in Escherichia coli Reviewed International journal

    Murakami K‡, Nasu H‡ (‡equally contribution), Fujiwara T, Takatsu N, Yoshida N, Furuta K, Kaito C

    J Bacteriol.   203 ( 12 )   e0051520   2021.5

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    DOI: 10.1128/JB.00515-20

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  • Total Syntheses and Chemical Biology Studies of Hymeglusin and Fusarilactone A, Novel Circumventors of β-Lactam Drug Resistance in Methicillin-Resistant Staphylococcus aureus Reviewed

    Kanaida M, Kimishima A, Eguchi S, Iwatsuki M, Watanabe Y, Honsho M, Hirose T, Noguchi Y, Nonaka K, Sennari G, Matsui H, Kaito C, Hanaki H, Asami Y, Sunazuka T

    ChemMedChem   16 ( 13 )   2106 - 2111   2021.3

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  • Proteolytic cleavage of HLA class II by human neutrophil elastase in pneumococcal pneumonia. Reviewed

    Domon H, Maekawa T, Isono T, Furuta K, Kaito C, Terao Y

    Scientific Reports   11 ( 1 )   2432   2021.1

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    DOI: 10.1038/s41598-021-82212-5

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  • Animal infection models using non-mammals Reviewed

    Kaito C, Murakami K, Imai L, Furuta K

    Microbiology and Immunology   64 ( 9 )   585 - 592: review   2020.8

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  • Non-pathogenic Escherichia coli acquires virulence by mutating a growth-essential LPS transporter. Reviewed

    Kaito C, Yoshikai H, Wakamatsu A, Miyashita A, Matsumoto Y, Fujiyuki T, Kato M, Ogura Y, Hayashi T, Isogai T, Sekimizu K

    PLOS Pathogens   16 ( 4 )   e1008469   2020.4

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  • Dataset for de novo transcriptome assembly of the African bullfrog Pyxicephalus adspersus Reviewed

    Yoshida N, Kaito C

    Data in Brief   6 ( 30 )   105388   2020.3

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  • Epidemiological study on the relationship between toxin production and psm‐mec mutations in MRSA isolates in Thailand Reviewed

    Tabuchi F, Lulitanond A, Lulitanond V, Thunyaharn S, Kaito C

    Microbiology and Immunology   64 ( 3 )   219 - 225   2020.3

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  • Tree of motility – A proposed history of motility systems in the tree of life Reviewed

    Makoto Miyata, Robert C. Robinson, Taro Q.P. Uyeda, Yoshihiro Fukumori, Shun ichi Fukushima, Shin Haruta, Michio Homma, Kazuo Inaba, Masahiro Ito, Chikara Kaito, Kentaro Kato, Tsuyoshi Kenri, Yoshiaki Kinosita, Seiji Kojima, Tohru Minamino, Hiroyuki Mori, Shuichi Nakamura, Daisuke Nakane, Koji Nakayama, Masayoshi Nishiyama, Satoshi Shibata, Katsuya Shimabukuro, Masatada Tamakoshi, Azuma Taoka, Yosuke Tashiro, Isil Tulum, Hirofumi Wada, Ken ichi Wakabayashi

    Genes to Cells   25 ( 1 )   6 - 21: review   2020.1

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    © 2020 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd Motility often plays a decisive role in the survival of species. Five systems of motility have been studied in depth: those propelled by bacterial flagella, eukaryotic actin polymerization and the eukaryotic motor proteins myosin, kinesin and dynein. However, many organisms exhibit surprisingly diverse motilities, and advances in genomics, molecular biology and imaging have showed that those motilities have inherently independent mechanisms. This makes defining the breadth of motility nontrivial, because novel motilities may be driven by unknown mechanisms. Here, we classify the known motilities based on the unique classes of movement-producing protein architectures. Based on this criterion, the current total of independent motility systems stands at 18 types. In this perspective, we discuss these modes of motility relative to the latest phylogenetic Tree of Life and propose a history of motility. During the ~4 billion years since the emergence of life, motility arose in Bacteria with flagella and pili, and in Archaea with archaella. Newer modes of motility became possible in Eukarya with changes to the cell envelope. Presence or absence of a peptidoglycan layer, the acquisition of robust membrane dynamics, the enlargement of cells and environmental opportunities likely provided the context for the (co)evolution of novel types of motility.

    DOI: 10.1111/gtc.12737

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  • Identification of 2H phosphoesterase superfamily proteins with 2'-CPDase activity Reviewed

    Mitsutomi S, Akimitsu N, Sekimizu K, Kaito C

    Biochimie   165   235 - 244   2019.8

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    DOI: 10.1016/j.biochi.2019.08.008

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  • Staphylococcus aureus aggregation in the plasma fraction of silkworm hemolymph Reviewed

    Ryuno H, Nigo F, Naguro I, Sekimizu K, Kaito C

    PLOS ONE   14 ( 5 )   e0217517   2019.5

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  • Two-spotted cricket as an animal infection model of human pathogenic fungi Reviewed

    Kochi Y, Matsumoto Y, Sekimizu K, Kaito C

    Drug Discoveries and Therapeutics   11 ( 5 )   259 - 266   2017.10

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    DOI: 10.5582/ddt.2017.01052

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  • Isolation of antibiotic-producing Pseudomonas species with low-temperature cultivation of temperate soil Reviewed

    Mitsutomi S, Sekimizu K, Kaito C

    Drug Discoveries and Therapeutics   11 ( 5 )   267 - 275   2017.10

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    DOI: 10.5582/ddt.2017.01053

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  • Impact of psm-mec in Methicillin-Resistant Staphylococcus aureus (ST764) Strains Isolated from Keratitis Patients Reviewed

    Suzuki T, Yamamoto T, Kaito C, Miyamoto H, Ohashi Y

    Microb Drug Resist.   22 ( 7 )   589 - 597   2016.10

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:MARY ANN LIEBERT, INC  

    Staphylococcus aureus is a predominant pathogen in keratitis, and the rate of methicillin-resistant S. aureus (MRSA) is increasing. In our previous study, genotypes of MRSA isolates from keratitis cases were classified into ST5 or ST764 lineage by multi-locus sequence typing. In this study, we examined the virulence properties of these MRSA keratitis isolates and its virulence determinants. There was no difference in the prevalence of virulence genes, such as adhesion and toxins, between ST5 and ST764 isolates. All ST5 isolates carried the intact psm-mec gene, which suppresses exotoxin production and colony spreading, but promotes biofilm formation. In contrast, all ST764 isolates had one point mutation in the psm-mec gene. Biofilm production in ST5 isolates was significantly higher than that in ST764 isolates, whereas colony spreading, hemolytic activity, and production of alpha-phenol-soluble modulins were higher in ST764 than in ST5 isolates. The toxicity of ST764 supernatants to corneal epithelial cells was higher than that of ST5 supernatants. These results suggest that the point mutation in the psm-mec gene contributes to the difference in virulence properties between ST5 and ST764 isolates in MRSA keratitis.

    DOI: 10.1089/mdr.2015.0315

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  • Cell-Surface Phenol Soluble Modulins Regulate Staphylococcus aureus Colony Spreading Reviewed

    Kizaki H, Omae Y, Tabuchi F, Saito Y, Sekimizu K, Kaito C

    PLOS ONE   11 ( 10 )   e0164523   2016.10

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    Staphylococcus aureus produces phenol-soluble modulins (PSMs), which are amphipathic small peptides with lytic activity against mammalian cells. We previously reported that PSM alpha 1-4 stimulate S. aureus colony spreading, the phenomenon of S. aureus colony expansion on the surface of soft agar plates, whereas delta-toxin (Hld, PSM gamma) inhibits colony-spreading activity. In this study, we revealed the underlying mechanism of the opposing effects of PSM alpha 1-4 and delta-toxin in S. aureus colony spreading. PSM alpha 1-4 and delta-toxin are abundant on the S. aureus cell surface, and account for 18% and 8.5% of the total amount of PSM alpha 1-4 and delta-toxin, respectively, in S. aureus overnight cultures. Knockout of PSM alpha 1-4 did not affect the amount of cell surface delta-toxin. In contrast, knockout of d-toxin increased the amount of cell surface PSM alpha 1-4, and decreased the amount of culture supernatant PSM alpha 1-4. The d-toxin inhibited PSM alpha 3 and PSM alpha 2 binding to the S. aureus cell surface in vitro. A double knockout strain of PSM alpha 1-4 and delta-toxin exhibited decreased colony spreading compared with the parent strain. Expression of cell surface PSM alpha 1-4, but not culture supernatant PSM alpha 1-4, restored the colony-spreading activity of the PSM alpha 1-4/delta-toxin double knockout strain. Expression of delta-toxin on the cell surface or in the culture supernatant did not restore the colony-spreading activity of the PSMa1-4/delta-toxin double knockout strain. These findings suggest that cell surface PSM alpha 1-4 promote S. aureus colony spreading, whereas delta-toxin suppresses colony-spreading activity by inhibiting PSM alpha 1-4 binding to the S. aureus cell surface.

    DOI: 10.1371/journal.pone.0164523

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  • Novel Nucleoside Diphosphatase Contributes to Staphylococcus aureus Virulence Reviewed

    Imae K, Saito Y, Kizaki H, Ryuno H, Mao H, Miyashita A, Suzuki Y, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   291 ( 36 )   18608 - 18619   2016.9

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    We identified SA1684 as a Staphylococcus aureus virulence gene using a silkworm infection model. The SA1684 gene product carried the DUF402 domain, which is found in RNA-binding proteins, and had amino acid sequence similarity with a nucleoside diphosphatase, Streptomyces coelicolor SC4828 protein. The SA1684-deletion mutant exhibited drastically decreased virulence, in which the LD50 against silkworms was more than 10 times that of the parent strain. The SA1684-deletion mutant also exhibited decreased exotoxin production and colony-spreading ability. Purified SA1684 protein had Mn2+ -or Co2+-dependent hydrolyzing activity against nucleoside diphosphates. Alanine substitutions of Tyr-88, Asp-106, and Asp-123/Glu-124, which are conserved between SA1684 and SC4828, diminished the nucleoside diphosphatase activity. Introduction of the wild-type SA1684 gene restored the hemolysin production of the SA1684-deletion mutant, whereas none of the alanine-substituted SA1684 mutant genes restored the hemolysin production. RNA sequence analysis revealed that SA1684 is required for the expression of the virulence regulatory genes agr, sarZ, and sarX, as well as metabolic genes involved in glycolysis and fermentation pathways. These findings suggest that the novel nucleoside diphosphatase SA1684 links metabolic-pathways and virulence gene expression and plays an important role in S. aureus virulence.

    DOI: 10.1074/jbc.M116.721845

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  • A human pathogenic bacterial infection model using the two-spotted cricket, Gryllus bimaculatus Reviewed

    Kochi Y, Miyashita A, Tsuchiya K, Mitsuyama M, Sekimizu K, Kaito C

    FEMS Microbiol Lett.   363 ( 15 )   fnw163   2016.8

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    Invertebrate animal species that can withstand temperatures as high as 37 degrees C, the human body temperature, are limited. In the present study, we utilized the two-spotted cricket, Gryllus bimaculatus, which lives in tropical and subtropical regions, as an animal model of human pathogenic bacterial infection. Injection of Pseudomonas aeruginosa or Staphylococcus aureus into the hemolymph killed crickets. Injected P. aeruginosa or S. aureus proliferated in the hemolymph until the cricket died. The ability of these pathogenic bacteria to kill the crickets was blocked by the administration of antibiotics. S. aureus gene-knockout mutants of virulence factors, including cvfA, agr and srtA, exhibited decreased killing ability compared with the parent strain. The dose at which 50% of crickets were killed by P. aeruginosa or S. aureus was not decreased at 37 degrees C compared with that at 27 degrees C. Injection of Listeria monocytogenes, which upregulates toxin expression at 37 degrees C, killed crickets, and the dose at which 50% of crickets were killed was decreased at 37 degrees C compared with that at 27 degrees C. These findings suggest that the two-spotted cricket is a useful model animal for evaluating the virulence properties of various human pathogenic bacteria at variable temperature including 37 degrees C.

    DOI: 10.1093/femsle/fnw163

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  • Body-enlarging effect of royal jelly in a non-holometabolous insect species, Gryllus bimaculatus Reviewed

    Miyashita A, Kizaki H, Sekimizu K, Kaito C

    Biology Open   5 ( 6 )   770 - 776   2016.6

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    Honeybee royal jelly is reported to have body-enlarging effects in holometabolous insects such as the honeybee, fly and silkmoth, but its effect in non-holometabolous insect species has not yet been examined. The present study confirmed the body-enlarging effect in silkmoths fed an artificial diet instead of mulberry leaves used in the previous literature. Administration of honeybee royal jelly to silkmoth from early larval stage increased the size of female pupae and adult moths, but not larvae (at the late larval stage) or male pupae. We further examined the body-enlarging effect of royal jelly in a non-holometabolous species, the two-spotted cricket Gryllus bimaculatus, which belongs to the evolutionarily primitive group Polyneoptera. Administration of royal jelly to G. bimaculatus from its early nymph stage enlarged both males and females at the mid-nymph and adult stages. In the cricket, the body parts were uniformly enlarged in both males and females; whereas the enlarged female silkmoths had swollen abdomens. Administration of royal jelly increased the number, but not the size, of eggs loaded in the abdomen of silkmoth females. In addition, fat body cells were enlarged by royal jelly in the silkmoth, but not in the cricket. These findings suggest that the body-enlarging effect of royal jelly is common in non-holometabolous species, G. bimaculatus, but it acts in a different manner than in holometabolous species.

    DOI: 10.1242/bio.019190

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  • Understanding of bacterial virulence using the silkworm infection model Reviewed

    Kaito C

    Drug Discoveries and Therapeutics   10 ( 1 )   30 - 33: review   2016.2

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    DOI: 10.5582/ddt.2016.01020

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  • No Effect of Body Size on the Frequency of Calling and Courtship Song in the Two-Spotted Cricket, Gryllus bimaculatus Reviewed

    Miyashita A, Kizaki H, Sekimizu K, Kaito C

    PLOS ONE   11 ( 1 )   e0146999   2016.1

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    The relationship between body size and vocalization parameters has been studied in many animal species. In insect species, however, the effect of body size on song frequency has remained unclear. Here we analyzed the effect of body size on the frequency spectra of mating songs produced by the two-spotted cricket, Gryllus bimaculatus. We recorded the calling songs and courtship songs of male crickets of different body sizes. The calling songs contained a frequency component that peaked at 5.7 kHz. On the other hand, courtship songs contained two frequency components that peaked at 5.8 and 14.7 kHz. The dominant frequency of each component in both the calling and courtship songs was constant regardless of body size. The size of the harp and mirror regions in the cricket forewings, which are the acoustic sources of the songs, correlated positively with body size. These findings suggest that the frequency contents of both the calling and courtship songs of the cricket are unaffected by whole body, harp, or mirror size.

    DOI: 10.1371/journal.pone.0146999

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  • Multidrug-Resistance Transporter AbcA Secretes Staphylococcus aureus Cytolytic Toxins Reviewed

    Yoshikai H, Kizaki H, Saito Y, Omae Y, Sekimizu K, Kaito C

    The Journal of Infectious Diseases   213 ( 2 )   295 - 304   2016.1

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    Phenol-soluble modulins (PSMs) are Staphylococcus aureus cytolytic toxins that lyse erythrocytes and neutrophils and have important functions in the S. aureus infectious process. The molecular mechanisms of PSM secretion, however, are not well understood. Here we report that knockout of the multidrug-resistance ABC transporter AbcA, which contributes to S. aureus resistance against antibiotics and chemicals, diminished the secreted amount of PSM, leading to the accumulation of PSM in the intracellular fraction. The amount of PSM in the culture supernatants of the abcA knockout mutants was restored by introduction of the wild-type abcA gene, whereas it was not completely restored by introduction of mutant abcA genes encoding AbcA mutant proteins carrying amino acid substitutions in the adenosine triphosphate binding motifs. The abcA knockout mutant exhibited attenuated virulence in a mouse systemic infection model. These findings suggest that the multidrug resistance transporter AbcA secretes PSMs and contributes to S. aureus virulence.

    DOI: 10.1093/infdis/jiv376

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  • 16S rRNA methyltransferase KsgA contributes to oxidative stress resistance and virulence in Staphylococcus aureus. Reviewed

    Kyuma T, Kizaki H, Ryuno H, Sekimizu K, Kaito C

    Biochimie   119   166 - 174   2015.12

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    DOI: 10.1016/j.biochi.2015.10.027

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  • Phenotypic and genomic comparisons of highly vancomycin-resistant Staphylococcus aureus strains developed from multiple clinical MRSA strains by in vitro mutagenesis Reviewed

    Ishii K, Tabuchi F, Matsuo M, Tatsuno K, Sato T, Okazaki M, Hamamoto H, Matsumoto Y, Kaito C, Aoyagi T, Hiramatsu K, Kaku M, Moriya K, Sekimizu K

    Scientific Reports   5   17092   2015.11

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    DOI: 10.1038/srep17092

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  • Ribosomal RNA methyltransferases contribute to Staphylococcus aureus virulence. Reviewed

    Kyuma T, Kimura S, Hanada Y, Suzuki T, Sekimizu K, Kaito C

    The FEBS journal   282 ( 13 )   2570 - 2584   2015.7

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    DOI: 10.1111/febs.13302

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  • Primed immune responses triggered by ingested bacteria lead to systemic infection tolerance in silkworms Reviewed

    Miyashita A, Takahashi S, Ishii K, Sekimizu K, Kaito C

    PLOS ONE   10 ( 6 )   e0130486   2015.6

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    DOI: 10.1371/journal.pone.0130486

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  • Current use of silkworm larvae (Bombyx mori) as an animal model in pharmaco-medical research Reviewed

    Nwibo DD, Hamamoto H, Matsumoto Y, Kaito C, Sekimizu K

    Drug Discoveries and Therapeutics   9 ( 2 )   133 - 135: review   2015.4

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    DOI: 10.5582/ddt.2015.01026

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  • Antibiotic-producing bacteria from stag beetle mycangia. Reviewed

    Miyashita A, Hirai Y, Sekimizu K, Kaito C

    Drug Discoveries and Therapeutics   9 ( 1 )   33 - 37   2015.2

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    DOI: 10.5582/ddt.2015.01000

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  • Impact of psm-mec in the mobile genetic element on the clinical characteristics and outcome of SCCmec-II methicillin-resistant Staphylococcus aureus bacteraemia in Japan Reviewed

    Aoyagi T, Kaito C, Sekimizu K, Omae Y, Saito Y, Mao H, Inomata S, Hatta M, Endo S, Kanamori H, Gu Y, Tokuda K, Yano H, Kitagawa M, Kaku M

    Clin Microbiol Infect.   20 ( 9 )   912 - 919   2014.9

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    Over-expression of alpha-phenol-soluble modulins (PSMs) results in high virulence of community-associated methicillin-resistant Staphylococcus aureus (MRSA). The psm-mec gene, located in the mobile genetic element SCCmec-II, suppresses PSMs production. Fifty-two patients with MRSA bacteraemia were enrolled. MRSA isolates were evaluated with regard to the psm-mec gene sequence, bacterial virulence, and the minimum inhibitory concentration (MIC) of vancomycin and teicoplanin. Fifty-one MRSA isolates were classified as SCCmec-II, and 10 had one point mutation in the psm-mec promoter. We compared clinical characteristics and outcomes between mutant MRSA and wild-type MRSA. Production of PSM3 in mutant MRSA was significantly increased, but biofilm formation was suppressed. Wild-type MRSA caused more catheter-related bloodstream infections (30/41 vs. 3/10, p0.0028), whereas mutant MRSA formed more deep abscesses (4/10 vs. 3/41, p0.035). Bacteraemia caused by mutant MRSA was associated with reduced 30-day mortality (1/10 vs. 13/41, p0.25), although this difference was not significant. The MIC90 of teicoplanin was higher for wild-type MRSA (1.5mg/L vs. 1mg/L), but the MIC of vancomycin was not different between the two groups. The 30-day mortality of MRSA with a high MIC of teicoplanin (1.5mg/L) was higher than that of strains with a lower MIC (0.75mg/L) (6/10 vs. 6/33, p0.017). Mutation of the psm-mec promoter contributes to virulence of SCCmec-II MRSA, and the product of psm-mec may determine the clinical characteristics of bacteraemia caused by SCCmec-II MRSA, but it does not affect mortality.

    DOI: 10.1111/1469-0691.12575

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  • Inhibition of Exotoxin Production by Mobile Genetic Element SCCmec-Encoded psm-mec RNA Is Conserved in Staphylococcal Species Reviewed

    Ikuo M, Nagano G, Saito Y, Mao H, Sekimizu K, Kaito C

    PLOS ONE   9 ( 6 )   e100260   2014.6

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    Staphylococcal species acquire antibiotic resistance by incorporating the mobile-genetic element SCCmec. We previously found that SCCmec-encoded psm-mec RNA suppresses exotoxin production as a regulatory RNA, and the psm-mec translation product increases biofilm formation in Staphylococcus aureus. Here, we examined whether the regulatory role of psm-mec on host bacterial virulence properties is conserved among other staphylococcal species, S. epidermidis and S. haemolyticus, both of which are important causes of nosocomial infections. In S. epidermidis, introduction of psm-mec decreased the production of cytolytic toxins called phenol-soluble modulins (PSMs) and increased biofilm formation. Introduction of psm-mec with a stop-codon mutation that did not express PSM-mec protein but did express psm-mec RNA also decreased PSM production, but did not increase biofilm formation. Thus, the psm-mec RNA inhibits PSM production, whereas the PSM-mec protein increases biofilm formation in S. epidermidis. In S. haemolyticus, introduction of psm-mec decreased PSM production, but did not affect biofilm formation. The mutated psm-mec with a stop-codon also caused the same effect. Thus, the psm-mec RNA also inhibits PSM production in S. haemolyticus. These findings suggest that the inhibitory role of psm-mec RNA on exotoxin production is conserved among staphylococcal species, although the stimulating effect of the psm-mec gene on biofilm formation is not conserved.

    DOI: 10.1371/journal.pone.0100260

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  • Identification of Staphylococcus aureus Colony-Spreading Stimulatory Factors from Mammalian Serum Reviewed

    Omae Y, Sekimizu K, Kaito C

    PLOS ONE   9 ( 5 )   e97670   2014.5

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    Staphylococcus aureus forms giant colonies on soft-agar surfaces, which is called colony-spreading. In the present study, we searched for host factors that influence S. aureus colony-spreading activity. The addition of calf serum, porcine serum, or silkworm hemolymph to soft-agar medium stimulated S. aureus colony-spreading activity. Gel filtration column chromatography of calf serum produced a high molecular weight fraction and a low molecular weight fraction, both of which exhibited colony-spreading stimulatory activity. In the low molecular weight fraction, we identified the stimulatory factor as bovine serum albumin. The stimulatory fraction in the high molecular weight fraction was identified as high-density lipoprotein (HDL) particles. Delipidation of HDL abolished the stimulatory activity of HDL. Phosphatidylcholine, which is the major lipid component in HDL particles, stimulated the colony-spreading activity. Other phosphatidylcholine-containing lipoprotein particles, low-density lipoprotein and very low-density lipoprotein, also showed colony-spreading stimulatory activity. These findings suggest that S. aureus colony-spreading activity is stimulated by albumin and lipoprotein particles in mammalian serum.

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  • Primed Immune Responses to Gram-negative Peptidoglycans Confer Infection Resistance in Silkworms Reviewed

    Miyashita A, Kizaki H, Kawasaki K, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   289 ( 20 )   14412 - 14421   2014.5

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    Background: Primed immune responses contribute to vertebrate host defense. Results: Silkworms acquire resistance to a pathogen by a preinjection of its heat-killed cells or its cell surface peptidoglycans. The amount of antimicrobial peptides is increased at the second round of infection. Conclusion: Invertebrates acquire infection resistance by peptidoglycan recognition and antimicrobial peptide increase. Significance: Molecular mechanisms of invertebrate primed immunity were revealed.
    A heightened immune response, in which immune responses are primed by repeated exposure to a pathogen, is an important characteristic of vertebrate adaptive immunity. In the present study, we examined whether invertebrate animals also exhibit a primed immune response. The LD50 of Gram-negative enterohemorrhagic Escherichia coli O157:H7 Sakai in silkworms was increased 100-fold by pre-injection of heat-killed Sakai cells. Silkworms pre-injected with heat-killed cells of a Gram-positive bacterium, Staphylococcus aureus, did not have resistance to Sakai. Silkworms preinjected with enterohemorrhagic E. coli peptidoglycans, cell surface components of bacteria, were resistant to Sakai infection. Silkworms preinjected with S. aureus peptidoglycans, however, were not resistant to Sakai. Silkworms preinjected with heat-killed Sakai cells showed persistent resistance to Sakai infection even after pupation. Repeated injection of heat-killed Sakai cells into the silkworms induced earlier and greater production of antimicrobial peptides than a single injection of heat-killed Sakai cells. These findings suggest that silkworm recognition of Gram-negative peptidoglycans leads to a primed immune reaction and increased resistance to a second round of bacterial infection.

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  • Western Blotting for Staphylococcus aureus AgrA Reviewed

    Kaito C, Sekimizu K

    Bio-protocol   4 ( 6 )   e1071   2014.3

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  • CvfA Protein and Polynucleotide Phosphorylase Act in an Opposing Manner to Regulate Staphylococcus aureus Virulence Reviewed

    Numata S, Nagata M, Mao H, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   289 ( 12 )   8420 - 8431   2014.3

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    Background: Production of 3-phosphorylated RNA by CvfA affects S. aureus virulence gene expression. Results: Disrupting pnpA-encoding exonuclease suppressed the cvfA-deleted mutant phenotype. Purified PNPase did not degrade 3-phosphorylated RNA. Conclusion: CvfA-produced 3-phosphorylated RNA inhibits PNPase-induced RNA degradation, resulting in hemolysin production by S. aureus. Significance: Altering the nucleotide structure at the RNA 3 terminus regulates S. aureus virulence.
    We previously identified CvfA (SA1129) as a Staphylococcus aureus virulence factor using a silkworm infection model. S. aureus cvfA-deleted mutants exhibit decreased expression of the agr locus encoding a positive regulator of hemolysin genes and decreased hemolysin production. CvfA protein hydrolyzes a 2,3-cyclic phosphodiester bond at the RNA 3 terminus, producing RNA with a 3-phosphate (3-phosphorylated RNA, RNA with a 3-phosphate). Here, we report that the cvfA-deleted mutant phenotype (decreased agr expression and hemolysin production) was suppressed by disrupting pnpA-encoding polynucleotide phosphorylase (PNPase) with 3- to 5-exonuclease activity. The suppression was blocked by introducing a pnpA-encoding PNPase with exonuclease activity but not by a pnpA-encoding mutant PNPase without exonuclease activity. Therefore, loss of PNPase exonuclease activity suppressed the cvfA-deleted mutant phenotype. Purified PNPase efficiently degraded RNA with 2,3-cyclic phosphate at the 3 terminus (2,3-cyclic RNA), but it inefficiently degraded 3-phosphorylated RNA. These findings indicate that 3-phosphorylated RNA production from 2,3-cyclic RNA by CvfA prevents RNA degradation by PNPase and contributes to the expression of agr and hemolysin genes. We speculate that in the cvfA-deleted mutant, 2,3-cyclic RNA is not converted to the 3-phosphorylated form and is efficiently degraded by PNPase, resulting in the loss of RNA essential for expressing agr and hemolysin genes, whereas in the cvfA/pnpA double-disrupted mutant, 2,3-cyclic RNA is not degraded by PNPase, leading to hemolysin production. These findings suggest that CvfA and PNPase competitively regulate RNA degradation essential for S. aureus virulence.

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  • Silkworm apolipophorin protein inhibits hemolysin gene expression of Staphylococcus aureus via binding to cell surface lipoteichoic acids Reviewed

    Omae Y, Hanada Y, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   288 ( 35 )   25542 - 50   2013.9

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    We previously reported that a silkworm hemolymph protein, apolipophorin (ApoLp), binds to the cell surface of Staphylococcus aureus and inhibits expression of the saePQRS operon encoding a two-component system, SaeRS, and hemolysin genes. In this study, we investigated the inhibitory mechanism of ApoLp on S. aureus hemolysin gene expression. ApoLp bound to lipoteichoic acids (LTA), an S. aureus cell surface component. The addition of purified LTA to liquid medium abolished the inhibitory effect of ApoLp against S. aureus hemolysin production. In an S. aureus knockdown mutant of ltaS encoding LTA synthetase, the inhibitory effects of ApoLp on saeQ expression and hemolysin production were attenuated. Furthermore, the addition of anti-LTA monoclonal antibody to liquid medium decreased the expression of S. aureus saeQ and hemolysin genes. In S. aureus strains expressing SaeS mutant proteins with a shortened extracellular domain, ApoLp did not decrease the saeQ expression. These findings suggest that ApoLp binds to LTA on the S. aureus cell surface and inhibits S. aureus hemolysin gene expression via a two-component regulatory system, SaeRS.

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  • Mobile Genetic Element SCCmec-encoded psm-mec RNA Suppresses Translation of agrA and Attenuates MRSA Virulence Reviewed

    Kaito C, Saito Y, Ikuo M, Omae Y, Mao H, Nagano G, Fujiyuki T, Numata S, Han X, Obata K, Hasegawa S, Yamaguchi H, Inokuchi K, Ito T, Hiramatsu K, Sekimizu K

    PLOS Pathogens   9 ( 4 )   e1003269   2013.4

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  • Lipopolysaccharide O-antigen of enterohemorrhagic Escherichia coli O157:H7 is required for killing both insects and mammals Reviewed

    Miyashita A, Iyoda S, Ishii K, Hamamoto H, Sekimizu K, Kaito C

    FEMS Microbiology Letters   333 ( 1 )   59 - 68   2012.8

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  • Inhibition of colony-spreading activity of Staphylococcus aureus by secretion of delta-hemolysin. Reviewed

    Omae Y, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   287 ( 19 )   15570 - 15579   2012.5

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  • Evaluation of Staphylococcus aureus virulence factors using a silkworm model. Reviewed

    Miyazaki S, Matsumoto Y, Sekimizu K, Kaito C

    FEMS Microbiology Letters   326 ( 2 )   116 - 124   2012.1

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  • Utilization of a silkworm model for understanding host-pathogen interactions Reviewed

    Kaito C, Yoshikai H, Sekimizu K

    Invertebrate Survival Journal   9   163 - 168: review   2012

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  • Silkworm Apolipophorin Protein Inhibits Staphylococcus aureus Virulence Reviewed

    Hanada Y, Sekimizu K, Kaito C

    The Journal of Biological Chemistry   286 ( 45 )   39360 - 39369   2011.11

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    Silkworm hemolymph inhibits hemolysin production by Staphylococcus aureus. We purified a factor in the silkworm hemolymph responsible for this inhibitory activity. The final fraction with the greatest specific activity contained 220- and 74-kDa proteins. Determination of the N-terminal amino acid sequence revealed that the 220- and 74-kDa proteins were apolipophorin I and apolipophorin II, respectively, indicating that the factor was apolipophorin (ApoLp). The purified ApoLp fraction showed decreased expression of S. aureus hla encoding alpha-hemolysin, hlb encoding beta-hemolysin, saeRS, and RNAIII, which activate the expression of these hemolysin genes. Injection of an anti-ApoLp antibody into the hemolymph increased the sensitivity of silkworms to the lethal effect of S. aureus. Hog gastric mucin, a mammalian homologue of ApoLp, decreased the expression of S. aureus hla and hlb. These findings suggest that ApoLp in the silkworm hemolymph inhibits S. aureus virulence and contributes to defense against S. aureus infection and that its activity is conserved in mammalian mucin.

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  • Digestion of extracellular DNA is required for giant colony formation of Staphylococcus aureus Reviewed

    Kaito C, Hirano T, Omae Y, Sekimizu K

    Microbial Pathogenesis   51 ( 3 )   142 - 148   2011.9

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    Staphylococcus aureus spreads on soft agar surfaces and forms giant colonies. Here, we examined the inhibitory role of extracellular DNA on the colony spreading activity. The double-deletion mutation of nuc1 and nuc2, which encode secretory nucleases, increased extracellular DNA and showed a decreased ability to form giant colonies. The addition of DNase I or micrococcal nuclease to the soft agar restored the ability of the nuc1-nuc2 double mutant to form giant colonies. In addition, the promoter activities of nuc1 and nuc2 in the wild-type strain were elevated in the peripheral region of the giant colony. These findings suggest that the digestion of extracellular DNA by secretory nucleases is required for the colony spreading activity of S. aureus. (C) 2011 Elsevier Ltd. All rights reserved.

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  • Sugar-responsive gene expression and the agr system are required for colony spreading in Staphylococcus aureus Reviewed

    Ueda T, Kaito C, Omae Y, Sekimizu K

    Microbial Pathogenesis   51 ( 3 )   178 - 185   2011.9

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    Staphylococcus aureus spreads on soft agar surfaces, which is called "colony spreading". Here, we report that the colony spreading in S. aureus was promoted by the addition of glucose to soft agar plates. Disruption of ccpA and hprK, which are involved in catabolite repression, decreased the colony spreading ability promoted by glucose. Deletion of the agr locus, a virulence regulatory element whose expression is activated by glucose in a ccpA-dependent manner, abolished the colony spreading promoted by glucose. Disruption of clpP and arlRS, which contributes to agr expression, also decreased glucose-promoted colony spreading. These findings suggest that S. aureus colony spreading requires the expression of agr, which is positively regulated by environmental carbon sources, and that virulence gene expression and colony spreading induced by agr are simultaneously activated in the S. aureus infectious process. (C) 2011 Elsevier Ltd. All rights reserved.

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  • Transcription and Translation Products of the Cytolysin Gene psm-mec on the Mobile Genetic Element SCCmec Regulate Staphylococcus aureus Virulence Reviewed

    Kaito C, Saito Y, Ikuo M, Omae Y, Mao H, Nagano G, Fujiyuki T, Numata S, Han X, Obata K, Hasegawa S, Yamaguchi H, Inokuchi K, Ito T, Hiramatsu K, Sekimizu K

    PLOS Pathogens   7 ( 2 )   e1001267   2011.2

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    The F region downstream of the mecI gene in the SCCmec element in hospital-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) contains two bidirectionally overlapping open reading frames (ORFs), the fudoh ORF and the psm-mec ORF. The psm-mec ORF encodes a cytolysin, phenol-soluble modulin (PSM)-mec. Transformation of the F region into the Newman strain, which is a methicillin-sensitive S. aureus (MSSA) strain, or into the MW2 (USA400) and FRP3757 (USA300) strains, which are community-acquired MRSA (CA-MRSA) strains that lack the F region, attenuated their virulence in a mouse systemic infection model. Introducing the F region to these strains suppressed colony-spreading activity and PSM alpha production, and promoted biofilm formation. By producing mutations into the psm-mec ORF, we revealed that (i) both the transcription and translation products of the psm-mec ORF suppressed colony-spreading activity and promoted biofilm formation; and (ii) the transcription product of the psm-mec ORF, but not its translation product, decreased PSM alpha production. These findings suggest that both the psm-mec transcript, acting as a regulatory RNA, and the PSM-mec protein encoded by the gene on the mobile genetic element SCCmec regulate the virulence of Staphylococcus aureus.

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  • Isolation of mammalian pathogenic bacteria using silkworms Reviewed

    Kaito C, Usui K, Kyuma T, Sekimizu K

    Drug Discoveries and Therapeutics   5 ( 2 )   66 - 70   2011

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  • The cvfC operon of Staphylococcus aureus contributes to virulence via expression of the thyA gene. Reviewed

    Ikuo M, Kaito C, Sekimizu K

    Microbial pathogenesis   49 ( 1-2 )   1 - 7   2010.7

  • Structure of a virulence regulatory factor CvfB reveals a novel winged helix RNA binding module. Reviewed

    Matsumoto Y, Xu Q, Miyazaki S, Kaito C, Farr CL, Axelrod HL, Chiu HJ, Klock HE, Knuth MW, Miller MD, Elsliger MA, Deacon AM, Godzik A, Lesley SA, Sekimizu K, Wilson IA

    Structure   18 ( 4 )   537 - 47   2010.3

  • Purification of a soil bacteria exotoxin using silkworm toxicity to measure specific activity. Reviewed

    Usui K, Miyazaki S, Kaito C, Sekimizu K

    Microbial pathogenesis   46 ( 2 )   59 - 62   2009.2

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  • A novel gene, fudoh, in the SCCmec region suppresses the colony spreading ability and virulence of Staphylococcus aureus Reviewed

    Kaito C, Omae Y, Matsumoto Y, Nagata M, Yamaguchi H, Aoto T, Ito T, Hiramatsu K, Sekimizu K

    PLOS ONE   3 ( 12 )   e3921   2008.12

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  • Gene Expression in Pre-MBT Embryos and Activation of Maternally-Inherited Program of Apoptosis to be Executed at around MBT as a Fail-Safe Mechanism in Xenopus Early Embryogenesis Reviewed

    Shiokawa K, Aso M, Kondo T, Uchiyama H, Kuroyanagi S, Takai J, Takahashi S, Kajitani M, Kaito C, Sekimizu K, Takayama E, Igarashi K, Hara H

    Gene Regulation and Systems Biology   29 ( 2 )   213 - 231   2008.5

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  • Phosphodiesterase activity of CvfA is required for virulence in Staphylococcus aureus. Reviewed

    Nagata M, Kaito C, Sekimizu K

    The Journal of Biological Chemistry   283 ( 4 )   2176 - 2184   2008.1

  • Regulation of exoprotein gene expression by the Staphylococcus aureus cvfB gene. Reviewed

    Matsumoto Y, Kaito C, Morishita D, Kurokawa K, Sekimizu K

    Infection and Immunity   75 ( 4 )   1964 - 1972   2007.4

  • Colony spreading in Staphylococcus aureus. Reviewed

    Kaito C, Sekimizu K

    Journal of Bacteriology   189 ( 6 )   2553 - 2557   2007.3

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  • Two-component signaling in the virulence of Staphylococcus aureus: a silkworm larvae-pathogenic agent infection model of virulence. Reviewed

    Kurokawa K, Kaito C, Sekimizu K

    Methods in Enzymology   422   233 - 44   2007

  • A silkworm model of pathogenic bacterial infection Reviewed

    Kaito C, Sekimizu K

    Drug Discoveries and Therapeutics   1 ( 2 )   89 - 93: review   2007

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  • Novel DNA binding protein SarZ contributes to virulence in Staphylococcus aureus. Reviewed

    Kaito C, Morishita D, Matsumoto Y, Kurokawa K, Sekimizu K

    Molecular Microbiology   62 ( 6 )   1601 - 1617   2006.12

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  • Use of silkworm larvae to study pathogenic bacterial toxins. Reviewed

    Hossain MS, Hamamoto H, Matsumoto Y, Razanajatovo IM, Larranaga J, Kaito C, Kasuga H, Sekimizu K

    The Journal of Biochemistry   140 ( 3 )   439 - 444   2006.9

  • Cleavage and survival of Xenopus embryos exposed to 8 T static magnetic fields in a rotating clinostat Reviewed

    Eguchi Y, Ueno S, Kaito C, Sekimizu K, Shiokawa K

    Bioelectromagnetics   27 ( 4 )   307 - 13   2006.5

  • Occurrence of pre-MBT synthesis of caspase-8 mRNA and activation of caspase-8 prior to execution of SAMDC (S-adenosylmethionine decarboxylase)-induced, but not p53-induced, apoptosis in Xenopus late blastulae. Reviewed

    Shiokawa K, Takayama E, Higo T, Kuroyanagi S, Kaito C, Hara H, Kajitani M, Sekimizu K, Tadakuma T, Miura K, Igarashi K, Yaoita Y

    Biochemical and Biophysical Research Communications   336 ( 2 )   682 - 691   2005.10

  • Silkworm pathogenic bacteria infection model for identification of novel virulence genes. Reviewed

    Kaito C, Kurokawa K, Matsumoto Y, Terao Y, Kawabata S, Hamada S, Sekimizu K

    Molecular Microbiology   56 ( 4 )   934 - 944   2005.5

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  • Participation of Rho-dependent transcription termination in oxidative stress sensitivity caused by an rpoB mutation. Reviewed

    Kawamura N, Kurokawa K, Ito T, Hamamoto H, Koyama H, Kaito C, Sekimizu K

    Genes to Cells   10 ( 5 )   477 - 487   2005.5

  • Quantitative evaluation of the therapeutic effects of antibiotics using silkworms infected with human pathogenic microorganisms Reviewed

    Hamamoto H, Kurokawa K, Kaito C, Kamura K, Manitra Razanajatovo I, Kusuhara H, Santa T, Sekimizu K

    Antimicrob Agents Chemother.   48 ( 3 )   774 - 779   2004.3

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    The injection of bacteria (Staphylococcus aureus, Stenotrophomonas maltophilia) or true fungi (Candida albicans, Candida tropicalis) that are pathogenic to humans into the silkworm hemolymph leads to death of the larvae within 2 days. Antibiotics used for clinical purposes have therapeutic effects on silkworms infected with these pathogens. The 50% effective doses obtained by injection into the silkworm hemolymph are consistent with those reported for mice. Injection of vancomycin and kanamycin into the silkworm hemolymph was effective, but oral administration was not. Chloramphenicol, which is effective by oral administration, appeared in the silkworm hemolymph soon after injection into the midgut, whereas vancomycin did not. Isolated midgut membranes were impermeable to vancomycin. Thus, the ineffectiveness of oral administration of vancomycin to silkworms is due to a lack of intestinal absorption.

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  • Inhibitory effects of basic or neutral phospholipid on acidic phospholipid-mediated dissociation of adenine nucleotide bound to DnaA protein, the initiator of chromosomal DNA replication Reviewed

    Ichihashi N, Kurokawa K, Matsuo M, Kaito C, Sekimizu K

    J Biol Chem.   278 ( 31 )   28778 - 28786   2003.8

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    DnaA protein activity, the initiator of chromosomal DNA replication in bacteria, is regulated by acidic phospholipids such as phosphatidylglycerol ( PG) or cardiolipin ( CL) via facilitation of the exchange reaction of bound adenine nucleotide. Total lipid isolated from exponentially growing Staphylococcus aureus cells facilitated the release of ATP bound to S. aureus DnaA protein, whereas that from stationary phase cells was inert. Fractionation of total lipid from stationary phase cells revealed that the basic phospholipid, lysylphosphatidylglycerol (LPG), inhibited PG- or CL-facilitated release of ATP from DnaA protein. There was an increase in LPG concentration during the stationary phase. A fraction of the total lipid from stationary phase cells of an integrational deletion mprF mutant, in which LPG was lost, facilitated the release of ATP from DnaA protein. A zwitterionic phospholipid, phosphatidylethanolamine, also inhibited PG- facilitated ATP release. These results indicate that interaction of DnaA protein with acidic phospholipids might be regulated by changes in the phospholipid composition of the cell membrane at different growth stages. In addition, the mprF mutant exhibited an increased amount of origin per cell in vivo, suggesting that LPG is involved in regulating the cell cycle event(s).

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  • Overexpression of S-adenosylmethionine decarboxylase (SAMDC) in Xenopus embryos activates maternal program of apoptosis as a "fail-safe" mechanism of early embryogenesis. Reviewed

    Kai M, Kaito C, Fukamachi H, Higo T, Takayama E, Hara H, Ohya Y, Igarashi K, Shiokawa K

    Cell Research   13 ( 3 )   147 - 58   2003.6

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  • Isolation and mutation site determination of the temperature-sensitive murB mutants of Staphylococcus aureus Reviewed

    Matsuo M, Kurokawa K, Nishida S, Li Y, Takimura H, Kaito C, Fukuhara N, Maki H, Miura K, Murakami K, Sekimizu K

    FEMS Microbiol Lett.   222 ( 1 )   107 - 113   2003.5

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    The murB gene encodes UDP-N-acetylenolpyruvylglucosamine reductase and functions in bacterial peptidoglycan biosynthesis. A plasmid carrying the murB gene restored the temperature-sensitive growth of six Staphylococcus aureus mutants, in which peptidoglycan biosynthesis stopped at a restrictive temperature. Specific activity of UDP-N-acetylenolpyruvylglucosamine reductase in extracts from the mutants was lower than that from wild-type cells. Nucleotide sequence determination revealed that each mutant had a single amino acid substitution in the mur-B gene and five of six mutations were located within domain 3, where the proposed substrate binding site is located. These results suggest that the murB gene is essential for growth of S. aureus and that domain 3 is important for the MurB activity. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

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  • Isolation and characterization of temperature-sensitive mutants of the Staphylococcus aureus dnaC gene. Reviewed

    Kaito C, Kurokawa K, Hossain MS, Akimitsu N, Sekimizu K

    FEMS Microbiol Lett.   210 ( 1 )   157 - 164   2002.4

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  • Silkworm larvae as an animal model of bacterial infection pathogenic to humans. Reviewed

    Kaito C, Akimitsu N, Watanabe H, Sekimizu K

    Microbial Pathogenesis   32 ( 4 )   183 - 90   2002.4

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  • Activation of the maternally preset program of apoptosis by microinjection of 5-aza-2'-deoxycytidine and 5-methyl-2'-deoxycytidine-5'-triphosphate in Xenopus laevis embryos. Reviewed

    Kaito C, Kai M, Higo T, Takayama E, Fukamachi H, Sekimizu K, Shiokawa K

    Development, Growth and Differentiation   43 ( 4 )   383 - 390   2001

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  • Whole genome sequencing of meticillin-resistant Staphylococcus aureus Reviewed

    Lancet   357 ( 9264 )   1225 - 1240   2001

  • Genetic identification of two distinct DNA polymerases, DnaE and PolC, that are essential for chromosomal DNA replication in Staphylococcus aureus Reviewed

    Molecular Genetics and Genomics   266 ( 4 )   564 - 571   2001

  • Overexpression of S-adenosylmethionine decarboxylase (SAMDC) activates the maternal program of apoptosis shortly after MBT in Xenopus embryos Reviewed

    Kai M, Higo T, Yokoska J, Kaito C, Kajita E, Fukamachi H, Takayama E, Igarashi K, Shiokawa K

    Int J Dev Biol.   44 ( 5 )   507 - 510   2000.8

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    Overexpression of S-adenosylmethionine decarboxylase (SAMDC) mRNA in 1- and 2-cell stage Xenopus embryos induces cell autonomous dissociation at the late blastula stage and developmental arrest at the early gastrula stage. The induction of cell dissociation took place "punctually" at the late blastula stage in the SAMDC-overexpressing cells, irrespective of the stage of the microinjection of SAMDC mRNA. When we examined the cells undergoing the dissociation, we found that they were TUNEL-positive and contained fragmented nuclei with condensed chromatin and fragmented DNA. Furthermore, by injecting Xenopus Bcl-2 mRNA together with SAMDC mRNA, we showed that SAMDC-overexpressing embryos are rescued completely by Bcl-2 and become tadpoles. These results indicate that cell dissociation induced by SAMDC overexpression is due to apoptotic cell death. Since the level of S-adenosylmethionine (SAM) is greatly reduced in SAMDC-overexpressing embryos and this induces inhibition of protein synthesis accompanied by the inhibition of DNA and RNA syntheses, we conclude that deficiency in SAM induced by SAMDC overexpression activates the maternal program of apoptosis in Xenopus embryos at the late blastula stage, but not before. We propose that this mechanism serves as a surveillance mechanism to check and eliminate cells physiologically damaged during the cleavage stage.

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  • Maternal program of apoptosis activated shortly after midblastula transition by overexpression of S-adenosylmethionine decarboxylase in Xenopus early embryos Reviewed

    Shiokawa K, Kai M, Higo T, Kaito C, Yokoska J, Yasuhiko Y, Kajita E, Nagano M, Yamada Y, Shibata M, Muto T, Shinga J, Hara H, Takayama E, Fukamachi H, Yaoita Y, Igarashi K

    Comp Biochem Physiol B Biochem Mol Biol.   126 ( 2 )   149 - 155: review   2000.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PERGAMON-ELSEVIER SCIENCE LTD  

    When we studied polyamine metabolism in Xenopus embryos, we cloned the cDNA for Xenopus S-adenosylmethionine decarboxylase (SAMDC), which converts SAM (S-adenosylmethionine), the methyl donor, into decarboxylated SAM (dcSAM), the aminopropyl donor, and microinjected its in vitro transcribed mRNA into Xenopus fertilized eggs. We found here that the mRNA injection induces a SAM deficient state in early embryos due to over-function of the overexpressed SAMDC, which in turn induces inhibition of protein synthesis. Such embryos developed quite normally until blastula stage, but stopped development at the early gastrula stage, due to induction of massive cell dissociation and cell autolysis, irrespective of the dosage and stage of the mRNA injection. We found that the dissociated cells were TUNEL-positive, contained fragmented nuclei with ladder-forming DNA, and furthermore, rescued completely by coinjection of Bcl-2 mRNA. Thus, overexpression of SAMDC in Xenopus embryos appeared to switch on apoptotic program, probably via inhibition of protein synthesis. Here, we briefly review our results together with those reported from other laboratories. After discussing the general importance of this newly discovered apoptotic program, we propose that the maternal program of apoptosis serves as a surveillance mechanism to eliminate metabolically severely-damaged cells and functions as a 'fail-safe' mechanism for normal development in Xenopus embryos. (C) 2000 Elsevier Science Inc. All rights reserved.

    DOI: 10.1016/S0305-0491(00)00193-0

    Web of Science

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Books

  • エンドソームと抗原提示

    古田 和幸、垣内 力( Role: Joint author)

    臨床免疫・アレルギー科 Vol.76 No.6 p622-628・科学評論社  2021.12 

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  • 進化実験による細菌病原性の理解

    垣内 力( Role: Sole author)

    羊土社・実験医学 増刊 39(2) p81-85「パンデミック時代の感染症研究 : 病原体の病原性、多様性、生活環から新型コロナウイルスを取り巻く社会の動きまで(編集:嘉糠洋陸)」  2021.2  ( ISBN:9784758103923

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    Total pages:209p   Language:Japanese

    CiNii Books

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  • 非ほ乳動物を用いた感染モデル

    Chikara Kaito( Role: Sole author)

    2017 

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  • 細菌間で保存された機能未知因子群からの新規病原性因子の同定

    Chikara Kaito( Role: Sole author)

    化学療法の領域 32(5):126-131. 医薬ジャーナル社  2016 

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  • 黄色ブドウ球菌の病原性制御機構に関する研究

    Chikara Kaito( Role: Sole author)

    日本細菌学雑誌 69(3):491-501. 日本細菌学会  2014 

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  • やさしい微生物学

    上原至雄, 垣内力, 北潔, 鈴木啓太郎, 関水和久, 辻勉, 細野哲司, 前田拓也, 牧純(編著者, 関水和久

    廣川書店  2011.4  ( ISBN:4567522109

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    Total pages:135  

    ASIN

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  • MRSAに見出された新規遺伝子「不動」

    Chikara Kaito

    化学療法の領域 25(8):46-53. 医薬ジャーナル社  2009 

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  • メチシリン耐性黄色ブドウ球菌のコロニースプレッディングおよび病原性発現調節を担う新規遺伝子「fudoh」

    Chikara Kaito

    感染・炎症・免疫 39(3):71-75. 医学の門社  2009 

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Industrial property rights

  • 病原性細菌の病原性を評価する方法、病原性の評価用キット及び病原性評価用遺伝子

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    Application no:特願2009-052647  Date applied:2009.2.17

    Announcement no:特開WO2009-154016 

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  • 感染症の予防又は治療のための薬剤及びその製造方法、評価方法及びスクリーニング方法、並びに、病原性細菌の病原異性の評価方法及び感染症の検査方法

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    Application no:特願2006-342713  Date applied:2006.12.20

    Announcement no:特開特開2008-148669 

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Awards

  • 小林六造記念賞

    2014.3   日本細菌学会  

    垣内 力

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Research Projects

  • 細菌の潜在的病原性をつかさどる分子基盤の解明

    Grant number:22H02869  2022 - 2024

    日本学術振興会  基盤研究(B) 

    垣内 力, 医歯薬学域

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    Authorship:Principal investigator 

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  • In vivo進化実験により病原性細菌出現の分子基盤に迫る

    Grant number:22K19435  2022 - 2023

    日本学術振興会 挑戦的研究(萌芽) 

    垣内 力,医歯薬学域

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    Authorship:Principal investigator 

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  • 多様なRNA相互作用因子を介したグラム陽性細菌の病原性制御機構の解明

    2019 - 2021

    日本学術振興会  基盤研究(B) 

    垣内 力, 医歯薬学系

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    Authorship:Principal investigator  Grant type:Competitive

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  • in vivo実験的進化系を利用した細菌の病原性システムの解明

    2019 - 2020

    日本学術振興会  挑戦的研究(萌芽) 

    垣内 力, 医歯薬学系

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    Authorship:Principal investigator  Grant type:Competitive

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  • グラム陽性細菌の病原性発現に必要なRNA相互作用分子群の探索とその分子機能の解明

    2019

    公益財団法人 武田科学振興財団  2019年度薬学系研究助成 

    垣内 力, 医歯薬学系

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    Authorship:Principal investigator 

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  • 熱帯性コオロギ感染モデルを用いた細菌の温度依存性病原性発動メカニズムの解明

    2016 - 2017

    日本学術振興会  Grant-in-Aid for Challenging Exploratory Research 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 治療効果を指向した新規抗菌薬の創出

    2015 - 2019

    日本学術振興会  基盤研究(S) 

    帝京大学, 関水 和久, 医真菌研究センター

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    Authorship:Coinvestigator(s)  Grant type:Competitive

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  • 世界の流行MRSAにおけるpsm-mec変異多型と病原性との関連性把握

    2015 - 2018

    日本学術振興会  Grant-in-Aid for Scientific Research (B)  基盤研究(B)

    垣内 力, 薬学系

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    Authorship:Principal investigator  Grant type:Competitive

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

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  • 新規RNA結合タンパク質群による細菌の病原性発現機構の解明

    2015 - 2017

    日本学術振興会  Grant-in-Aid for Scientific Research (B)  基盤研究(B)

    垣内 力, 薬学系

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\6500000 ( Direct expense: \5000000 、 Indirect expense:\1500000 )

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  • 食用キノコに常在する微生物の生理的機能に関する研究

    2015

    薬学振興会(公財)  基礎的研究助成 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 遺伝子組み換え常在細菌を利用した動物に対する成長ホルモン供給法の確立

    2014 - 2015

    日本学術振興会  Grant-in-Aid for Challenging Exploratory Research  挑戦的萌芽研究

    垣内 力, 薬学系

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3640000 ( Direct expense: \2800000 、 Indirect expense:\840000 )

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  • 新規RNA 結合蛋白質群による細菌の病原性調節機構の理解

    2014

    かなえ医薬振興財団  研究助成金 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 黄色ブドウ球菌の細胞外毒素分泌トランスポーターの同定

    2014

    野田産業科学研究所(公財)  野田産研研究助成 奨励研究助成 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 黄色ブドウ球菌の新規移動様式の分子機構

    2013 - 2014

    日本学術振興会  新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    垣内 力, 薬学系

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\10270000 ( Direct expense: \7900000 、 Indirect expense:\2370000 )

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  • 大腸菌の高病原性化をひきおこす分子機構の解明

    2013

    薬学研究奨励財団  研究助成金 グループA 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 細菌の潜在的病原性を規定する分子機構の解明

    2013

    持田記念医学薬学振興財団  研究助成金 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 外来遺伝子によるMRSAの病原性調節機構に関する研究

    2012 - 2014

    日本学術振興会  基盤研究(C)  基盤研究(C)

    垣内力, 薬学系

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5460000 ( Direct expense: \4200000 、 Indirect expense:\1260000 )

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  • カイコ感染モデルを利用した宿主—病原体相互作用の理解

    2011 - 2013

    日本学術振興会  基盤研究(A) 

    関水 和久, 薬学系, 教授

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    Grant type:Competitive

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  • FudohによるMRSAの病原性制御

    2009

    内藤記念科学振興財団  内藤記念科学奨励金 

    垣内 力

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3000000

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  • カイコ感染モデルを基盤とした細菌の病原性発現システムの解明

    2008 - 2010

    日本学術振興会  基盤研究(B) 

    関水 和久, 薬学系, 教授

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    Grant type:Competitive

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  • Research on virulence regulatory mechanisms by CVF

    Grant number:20790057  2008 - 2009

    Ministry of Education, Culture, Sports, Science and Technology  Grants-in-Aid for Scientific Research(若手研究(B))  若手研究(B)

    Chikara KAITO

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    Authorship:Principal investigator  Grant type:Competitive

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

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  • カイコ感染モデルを用いたグラム陽性細菌に保存された病原性発現機構の解明

    2007 - 2008

    日本学術振興会  特定領域研究 

    関水 和久, 薬学系, 教授

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    Grant type:Competitive

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  • 黄色ブドウ球菌の滑走能低下を指標とした抗菌治療薬に関する研究

    2007

    日本学術振興会  萌芽研究 

    関水 和久, 薬学系, 教授

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    Grant type:Competitive

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  • 細胞間で保存された新規病原性遺伝子の同定

    2006 - 2007

    日本学術振興会  若手研究(B)  若手研究(B)

    垣内 力, 薬学系, 助手

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\3500000 ( Direct expense: \3500000 )

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  • カイコ感染モデルを用いた新規病原性遺伝子の同定

    2006

    JST  シーズ発掘試験研究 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 黄色ブドウ球菌の滑走能を規定する遺伝子群の探索

    2006

    薬学振興会(公財)  基礎的研究助成 

    Chikara Kaito

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    Authorship:Principal investigator  Grant type:Competitive

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  • 黄色ブドウ球菌の宿主相互作用因子のカイコ感染モデルを用いた網羅的同定と機能解析

    2002 - 2004

    日本学術振興会  特別研究員奨励費(DC1) 

    垣内 力

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    Grant type:Competitive

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Class subject in charge

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  • 高等学校 授業 研究室見学(科研費アウトリーチ等)

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    宮城県、埼玉県、三重県、静岡県、広島県、岡山県、島根県 

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  • 墨田区医師会立看護専門学校 生化学講義(地域医療人材育成)

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