start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=1 article-no= start-page=9 end-page=18 dt-received= dt-revised= dt-accepted= dt-pub-year=2013 dt-pub=201302 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Phospholipase C Produced by Clostridium botulinum Types C and D:Comparison of Gene, Enzymatic, and Biological Activities with Those of Clostridium perfringens Alpha-toxin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs), the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin. Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity;however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival. en-copyright= kn-copyright= en-aut-name=FatmawatiNi Nengah Dwi en-aut-sei=Fatmawati en-aut-mei=Ni Nengah Dwi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SakaguchiYoshihiko en-aut-sei=Sakaguchi en-aut-mei=Yoshihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SuzukiTomonori en-aut-sei=Suzuki en-aut-mei=Tomonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OdaMasataka en-aut-sei=Oda en-aut-mei=Masataka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ShimizuKenta en-aut-sei=Shimizu en-aut-mei=Kenta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=YamamotoYumiko en-aut-sei=Yamamoto en-aut-mei=Yumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SakuraiJun en-aut-sei=Sakurai en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MatsushitaOsamu en-aut-sei=Matsushita en-aut-mei=Osamu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=OgumaKeiji en-aut-sei=Oguma en-aut-mei=Keiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=2 en-affil= kn-affil=Interdisciplinary Research Organization, Miyazaki University affil-num=3 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=4 en-affil= kn-affil=Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University affil-num=5 en-affil= kn-affil=Shin-Nakamura Chemical Co., Ltd affil-num=6 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=7 en-affil= kn-affil=Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University affil-num=8 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=9 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences en-keyword=botulinum phospholipase C kn-keyword=botulinum phospholipase C en-keyword=botulinum toxin kn-keyword=botulinum toxin en-keyword=phospholipase C activity kn-keyword=phospholipase C activity en-keyword=sphingomyelinase activity kn-keyword=sphingomyelinase activity en-keyword=hemolytic activity kn-keyword=hemolytic activity END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=4 article-no= start-page=291 end-page=297 dt-received= dt-revised= dt-accepted= dt-pub-year=2012 dt-pub=201208 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Intraprostatic Botulinum Neurotoxin Type A Injection for Benign Prostatic Hyperplasia:Preliminary Results with a Newly Purified Neurotoxin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Several studies have demonstrated the efficacy of intraprostatic injection of botulinum neurotoxin type A (BoNT/A) against symptomatic benign prostatic hyperplasia (BPH). The most commonly used BoNT/A product, Botox®, forms large complexes and composed of neurotoxin (NTX) as well as non-toxic components. We purified NTX lacking non-toxic components. We investigated the efficacy of this newly purified NTX for men with BPH. Ten male patients (mean age, 70.0 years) with BPH received 100 units (prostate volume [PV] <30ml) or 200 units (PV ァ30ml) of NTX injected into the prostate via a minimally invasive outpatient technique. Evaluation included uroflowmetry, postvoid residual urine volume (PVR), PV, and International Prostate Symptom Score (IPSS) measured at baseline and 1, 3, 6, and 12 months post-treatment. The status of 7 of the 10 patients examined was found to have improved within 1 month of treatment. The mean IPSS decreased from 23.8±7.0 to 16.3±10.3 (p=0.0093) at 1 month, to 14.9±8.2 (p=0.0074) at 3 months, and to 16.9±7.3 (p=0.018) at 12 months. The mean PV decreased from 47.8±21.2 to 39.2±19.5ml (p=0.0076) at 3 months. The PVR improved at 3 and 6 months post-treatment. Intraprostatic NTX injection induces prostate shrinkage and is effective in men with BPH. en-copyright= kn-copyright= en-aut-name=YokoyamaTeruhiko en-aut-sei=Yokoyama en-aut-mei=Teruhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamamotoYumiko en-aut-sei=Yamamoto en-aut-mei=Yumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SuzukiTomonori en-aut-sei=Suzuki en-aut-mei=Tomonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OgumaKeiji en-aut-sei=Oguma en-aut-mei=Keiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=NagaiAtsushi en-aut-sei=Nagai en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Department of Urology, Kawasaki Medical School affil-num=2 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science affil-num=3 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science affil-num=4 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science affil-num=5 en-affil= kn-affil=Department of Urology, Kawasaki Medical School en-keyword=botulinum neurotoxin type A kn-keyword=botulinum neurotoxin type A en-keyword=benign prostatic hyperplasia kn-keyword=benign prostatic hyperplasia en-keyword=therapy kn-keyword=therapy END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=3 article-no= start-page=253 end-page=261 dt-received= dt-revised= dt-accepted= dt-pub-year=2012 dt-pub=201206 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Clostridium botulinum Type E Toxins Bind to Caco-2 Cells by a Different Mechanism from That of Type A Toxins en-subtitle= kn-subtitle= en-abstract= kn-abstract=Cultured Clostridium botulinum strains produce progenitor toxins designated as 12S, 16S, and 19S toxins. The 12S toxin consists of a neurotoxin (NTX, 7S) and a non-toxic non-hemagglutinin (NTNH). The 16S and 19S toxins are formed by conjugation of the 12S toxin with hemagglutinin (HA), and the 19S toxin is a dimer of the 16S toxin. Type A cultures produce all 3 of these progenitor toxins, while type E produces only the 12S toxin. The 7S toxin is cleaved into heavy (H) and light (L) chains by a protease(s) in some strains, and the H chain has 2 domains, the N-terminus (Hn) and C-terminus (Hc). It has been reported that type A toxins bind to the intestinal cells or cultured cells via either HA or Hc. In this study, we investigated the binding of type A and E toxins to Caco-2 cells using Western blot analysis. Both the type E 7S and 12S toxins bound to the cells, with the 7S toxin binding more strongly, whereas, in the type A strain, only the 16S/19S toxins showed obvious binding. Pre-incubation of the type E 7S toxin with IgG against recombinant type E Hc significantly inhibited the 7S toxin binding, indicating that Hc might be a main binding domain of the type E toxin. en-copyright= kn-copyright= en-aut-name=ZhangKai en-aut-sei=Zhang en-aut-mei=Kai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamamotoYumiko en-aut-sei=Yamamoto en-aut-mei=Yumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SuzukiTomonori en-aut-sei=Suzuki en-aut-mei=Tomonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YokotaKenji en-aut-sei=Yokota en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MaShaobo en-aut-sei=Ma en-aut-mei=Shaobo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=Nengah Dwi FatmawatiNi en-aut-sei=Nengah Dwi Fatmawati en-aut-mei=Ni kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=OgumaKeiji en-aut-sei=Oguma en-aut-mei=Keiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine affil-num=2 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine affil-num=3 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine affil-num=4 en-affil= kn-affil=Graduate School of Health Sciences, Okayama University affil-num=5 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine affil-num=6 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine affil-num=7 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine en-keyword=Clostridum botulinum kn-keyword=Clostridum botulinum en-keyword=neurotoxins kn-keyword=neurotoxins en-keyword=Caco-2 kn-keyword=Caco-2 en-keyword=binding kn-keyword=binding en-keyword=Hc kn-keyword=Hc END