start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=1 article-no= start-page=10 end-page=14 dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20211021 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Attempt of thyX gene silencing and construction of a thyX deleted clone in a Mycobacterium bovis BCG en-subtitle= kn-subtitle= en-abstract= kn-abstract=Mycobacterium tuberculosis, the causative agent of tuberculosis, possess flavin-dependent thymidylate synthase, ThyX. Since thyX is absent in humans and was shown to be essential for M. tuberculosis normal growth, ThyX is thought to be an attractive novel TB drug target. This study assessed thyX essentiality in Mycobacterium bovis BCG strains using CRISPR interference based gene silencing and found that thyX is not essential in an M. bovis BCG Tokyo derivative strain. A thyX deletion mutant strain was successfully constructed from that strain, which reinforces the non-essentiality of thyX under a certain genetic background. en-copyright= kn-copyright= en-aut-name=ArimuraYuki en-aut-sei=Arimura en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MinatoYusuke en-aut-sei=Minato en-aut-mei=Yusuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WadaTakayuki en-aut-sei=Wada en-aut-mei=Takayuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=NakayamaMasaaki en-aut-sei=Nakayama en-aut-mei=Masaaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=RyumonAyako en-aut-sei=Ryumon en-aut-mei=Ayako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HirataNao en-aut-sei=Hirata en-aut-mei=Nao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NakajimaChie en-aut-sei=Nakajima en-aut-mei=Chie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=SuzukiYasuhiko en-aut-sei=Suzuki en-aut-mei=Yasuhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=AtoManabu en-aut-sei=Ato en-aut-mei=Manabu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=KobayashiKazuo en-aut-sei=Kobayashi en-aut-mei=Kazuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=OharaNaoko en-aut-sei=Ohara en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=IidaSeiji en-aut-sei=Iida en-aut-mei=Seiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=OharaNaoya en-aut-sei=Ohara en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= affil-num=1 en-affil=Department of Oral and Maxillofacial Reconstructive Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Microbiology, Fujita Health University School of Medicine kn-affil= affil-num=3 en-affil=Department of Microbiology, Graduate School of Human Life Science, Osaka City University kn-affil= affil-num=4 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Microbiology, Fujita Health University School of Medicine kn-affil= affil-num=7 en-affil=Division of Bioresources, Hokkaido University International Institute for Zoonosis Control kn-affil= affil-num=8 en-affil=Division of Bioresources, Hokkaido University International Institute for Zoonosis Control kn-affil= affil-num=9 en-affil=Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases kn-affil= affil-num=10 en-affil=Department of Immunology, National Institute of Infectious Diseases kn-affil= affil-num=11 en-affil=Department of Operative Dentistry, Okayama University Hospital, Okayama University kn-affil= affil-num=12 en-affil=Department of Oral and Maxillofacial Reconstructive Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=13 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=thymidylate synthase kn-keyword=thymidylate synthase en-keyword=ThyX kn-keyword=ThyX en-keyword=Mycobacterium kn-keyword=Mycobacterium en-keyword=BCG kn-keyword=BCG END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page=e13312 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=202153 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Roles of Porphyromonas gulae proteases in bacterial and host cell biology en-subtitle= kn-subtitle= en-abstract= kn-abstract=Porphyromonas gulae, an animal-derived periodontal pathogen, expresses several virulence factors, including fimbria, lipopolysaccharide (LPS) and proteases. We previously reported that its invasive efficiency was dependent on fimbriae types. In addition, P. gulae LPS increased inflammatory responses via toll-like receptors. The present study was conducted to investigate the involvement of P. gulae proteases in bacterial and host cell biology. Porphyromonas gulae strains showed an ability to agglutinate mouse erythrocytes and also demonstrated co-aggregation with Actinomyces viscosus, while the protease inhibitors antipain, PMSF, TLCK and leupeptin diminished P. gulae proteolytic activity, resulting in inhibition of haemagglutination and co-aggregation with A. viscosus. In addition, specific proteinase inhibitors were found to reduce bacterial cell growth. Porphyromonas gulae inhibited Ca9-22 cell proliferation in a multiplicity of infection- and time-dependent manner. Additionally, P. gulae-induced decreases in cell contact and adhesion-related proteins were accompanied by a marked change in cell morphology from well spread to rounded. In contrast, inhibition of protease activity prevented degradation of proteins, such as E-cadherin, beta-catenin and focal adhesion kinase, and also blocked inhibition of cell proliferation. Together, these results indicate suppression of the amount of human proteins, such as gamma-globulin, fibrinogen and fibronectin, by P. gulae proteases, suggesting that a novel protease complex contributes to bacterial virulence. en-copyright= kn-copyright= en-aut-name=UrmiAlam Saki en-aut-sei=Urmi en-aut-mei=Alam Saki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=InabaHiroaki en-aut-sei=Inaba en-aut-mei=Hiroaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NomuraRyota en-aut-sei=Nomura en-aut-mei=Ryota kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YoshidaShoko en-aut-sei=Yoshida en-aut-mei=Shoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OharaNaoya en-aut-sei=Ohara en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=AsaiFumitoshi en-aut-sei=Asai en-aut-mei=Fumitoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NakanoKazuhiko en-aut-sei=Nakano en-aut-mei=Kazuhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=Matsumoto‐NakanoMichiyo en-aut-sei=Matsumoto‐Nakano en-aut-mei=Michiyo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Department of Pediatric Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Pediatric Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Pediatric Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Pediatric Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Oral Microbiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences and the Advanced Research Center for Oral and Craniofacial Sciences, Dental School, Okayama University kn-affil= affil-num=6 en-affil=Department of Pharmacology, School of Veterinary Medicine Azabu University kn-affil= affil-num=7 en-affil=Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry kn-affil= affil-num=8 en-affil=Department of Pediatric Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=coaggregation kn-keyword=coaggregation en-keyword=haemagglutination kn-keyword=haemagglutination en-keyword=P. gulae kn-keyword=P. gulae en-keyword=protease kn-keyword=protease en-keyword=protein degradation kn-keyword=protein degradation END start-ver=1.4 cd-journal=joma no-vol=62 cd-vols= no-issue=4 article-no= start-page=322 end-page=326 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=202012 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Construction and characterization of the PGN_0296 mutant of Porphyromonas gingivalis en-subtitle= kn-subtitle= en-abstract= kn-abstract=The periodontal pathogen Porphyromonas gingivalis produces gingipains (Kgp, RgpA, and RgpB), cysteine proteases involved in the organism's virulence, and pigmentation. We previously showed that deletion of the PGN_0297 and PGN_0300 genes reduced the proteolytic activity of gingipains. The role of the PGN_0296 gene, consisting of an operon with the PGN_0297 and PGN_0300 genes, is unclear. Herein, we examined the effect of PGN_0296 gene deletion on the proteolytic activity. Although the proteolytic activity of the gingipains did not decrease in the culture supernatant of a PGN_0296 gene deletion mutant (ΔPGN_0296), the growth was delayed. en-copyright= kn-copyright= en-aut-name=ShahriarAbu Saleh Muhammad en-aut-sei=Shahriar en-aut-mei=Abu Saleh Muhammad kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OnoShintaro en-aut-sei=Ono en-aut-mei=Shintaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NakayamaMasaaki en-aut-sei=Nakayama en-aut-mei=Masaaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OharaNaoko en-aut-sei=Ohara en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OharaNaoya en-aut-sei=Ohara en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Periodontal Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Operative Dentistry, Okayama University Hospital, Okayama University kn-affil= affil-num=5 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=Porphyromonas gingivalis kn-keyword=Porphyromonas gingivalis en-keyword=Periodontal diseases kn-keyword=Periodontal diseases en-keyword=Gingipain kn-keyword=Gingipain en-keyword=Bacterial secretion system kn-keyword=Bacterial secretion system END start-ver=1.4 cd-journal=joma no-vol=28 cd-vols= no-issue=1 article-no= start-page=100 end-page=118 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200108 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Rescue from Stx2-Producing E. coli-Associated Encephalopathy by Intravenous Injection of Muse Cells in NOD-SCID Mice en-subtitle= kn-subtitle= en-abstract= kn-abstract=Shiga toxin-producing Escherichia coli (STEC) causes hemorrhagic colitis, hemolytic uremic syndrome, and acute encephalopathies that may lead to sudden death or severe neurologic sequelae. Current treatments, including immunoglobulin G (IgG) immunoadsorption, plasma exchange, steroid pulse therapy, and the monoclonal antibody eculizumab, have limited effects against the severe neurologic sequelae. Multilineage-differentiating stress-enduring (Muse) cells are endogenous reparative non-tumorigenic stem cells that naturally reside in the body and are currently under clinical trials for regenerative medicine. When administered intravenously, Musecells accumulate to the damaged tissue, where they exert anti-inflammatory, anti-apoptotic, anti-fibrotic, and immunomodulatory effects, and replace damaged cells by differentiating into tissue-constituent cells. Here, severely immunocompromised non-obese diabetic/severe combined immunodeficiency (NOD-SCID) mice orally inoculated with 9 × 109 colony-forming units of STEC O111 and treated 48 h later with intravenous injection of 5 × 104 Muse cells exhibited 100% survival and no severe after-effects of infection. Suppression of granulocyte-colony-stimulating factor (G-CSF) by RNAi abolished the beneficial effects of Muse cells, leading to a 40% death and significant body weight loss, suggesting the involvement of G-CSF in the beneficial effects of Muse cells in STEC-infected mice. Thus, intravenous administration of Muse cells could be a candidate therapeutic approach for preventing fatal encephalopathy after STEC infection. en-copyright= kn-copyright= en-aut-name=OzuruRyo en-aut-sei=Ozuru en-aut-mei=Ryo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=WakaoShohei en-aut-sei=Wakao en-aut-mei=Shohei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TsujiTakahiro en-aut-sei=Tsuji en-aut-mei=Takahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OharaNaoya en-aut-sei=Ohara en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MatsubaTakashi en-aut-sei=Matsuba en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=AmuranMuhammad Y. en-aut-sei=Amuran en-aut-mei=Muhammad Y. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=IsobeJunko en-aut-sei=Isobe en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=IinoMorio en-aut-sei=Iino en-aut-mei=Morio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=NishidaNaoki en-aut-sei=Nishida en-aut-mei=Naoki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=MatsumotoSari en-aut-sei=Matsumoto en-aut-mei=Sari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=IwadateKimiharu en-aut-sei=Iwadate en-aut-mei=Kimiharu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=KonishiNoriko en-aut-sei=Konishi en-aut-mei=Noriko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=YasudaKaori en-aut-sei=Yasuda en-aut-mei=Kaori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=TashiroKosuke en-aut-sei=Tashiro en-aut-mei=Kosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=HidaMisato en-aut-sei=Hida en-aut-mei=Misato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= en-aut-name=YadoiwaArisato en-aut-sei=Yadoiwa en-aut-mei=Arisato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=16 ORCID= en-aut-name=KatoShinsuke en-aut-sei=Kato en-aut-mei=Shinsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=17 ORCID= en-aut-name=YamashitaEijiro en-aut-sei=Yamashita en-aut-mei=Eijiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=18 ORCID= en-aut-name=MatsumotoSohkichi en-aut-sei=Matsumoto en-aut-mei=Sohkichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=19 ORCID= en-aut-name=KurozawaYoichi en-aut-sei=Kurozawa en-aut-mei=Yoichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=20 ORCID= en-aut-name=DezawaMari en-aut-sei=Dezawa en-aut-mei=Mari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=21 ORCID= en-aut-name=FujiiJun en-aut-sei=Fujii en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=22 ORCID= affil-num=1 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= affil-num=2 en-affil=Department of Stem Cell Biology and Histology, Graduate School of Medicine, Tohoku University kn-affil= affil-num=3 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= affil-num=4 en-affil=Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= affil-num=6 en-affil=Department of Neurology, Hasanuddin University Faculty of Medicine kn-affil= affil-num=7 en-affil=Department of Bacteriology, Toyama Institute of Health kn-affil= affil-num=8 en-affil=Division of Legal Medicine, Department of Social Medicine, Faculty of Medicine, Tottori University kn-affil= affil-num=9 en-affil=Department of Legal Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama kn-affil= affil-num=10 en-affil=Department of Forensic Medicine, The Jikei University School of Medicine kn-affil= affil-num=11 en-affil=Department of Forensic Medicine, The Jikei University School of Medicine kn-affil= affil-num=12 en-affil=Department of Food Microbiology, Tokyo Metropolitan Institute of Public kn-affil= affil-num=13 en-affil=Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University kn-affil= affil-num=14 en-affil=Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University kn-affil= affil-num=15 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= affil-num=16 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= affil-num=17 en-affil=Division of Neuropathology, Department of Brain and Neuroscience, Faculty of Medicine, School of Medicine, Tottori University Faculty of Medicine kn-affil= affil-num=18 en-affil=Division of Clinical Radiology, Tottori University Hospital kn-affil= affil-num=19 en-affil=Department of Bacteriology, Niigata University School of Medicine kn-affil= affil-num=20 en-affil=Division of Health Administration and Promotion, Department of Social Medicine, Faculty of Medicine, Tottori University kn-affil= affil-num=21 en-affil=Department of Stem Cell Biology and Histology, Graduate School of Medicine, Tohoku University kn-affil= affil-num=22 en-affil=Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University kn-affil= en-keyword=Muse cells kn-keyword=Muse cells en-keyword=Shiga toxin-producing Escherichia coli kn-keyword=Shiga toxin-producing Escherichia coli en-keyword=acute encephalopathy kn-keyword=acute encephalopathy END