start-ver=1.4 cd-journal=joma no-vol=23 cd-vols= no-issue=5 article-no= start-page=209 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250514 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Novel Anti-MRSA Peptide from Mangrove-Derived Virgibacillus chiguensis FN33 Supported by Genomics and Molecular Dynamics en-subtitle= kn-subtitle= en-abstract= kn-abstract=Antimicrobial resistance (AMR) is a global health threat, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the major resistant pathogens. This study reports the isolation of a novel mangrove-derived bacterium, Virgibacillus chiguensis FN33, as identified through genome analysis and the discovery of a new anionic antimicrobial peptide (AMP) exhibiting anti-MRSA activity. The AMP was composed of 23 amino acids, which were elucidated as NH3-Glu-Gly-Gly-Cys-Gly-Val-Asp-Thr-Trp-Gly-Cys-Leu-Thr-Pro-Cys-His-Cys-Asp-Leu-Phe-Cys-Thr-Thr-COOH. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for MRSA were 8 ?g/mL and 16 ?g/mL, respectively. FN33 AMP induced cell membrane permeabilization, suggesting a membrane-disrupting mechanism. The AMP remained stable at 30?40 ‹C but lost activity at higher temperatures and following exposure to proteases, surfactants, and extreme pH. All-atom molecular dynamics simulations showed that the AMP adopts a ƒΐ-sheet structure upon membrane interaction. These findings suggest that Virgibacillus chiguensis FN33 is a promising source of novel antibacterial agents against MRSA, supporting alternative strategies for drug-resistant infections. en-copyright= kn-copyright= en-aut-name=SermkaewNamfa en-aut-sei=Sermkaew en-aut-mei=Namfa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AtipairinApichart en-aut-sei=Atipairin en-aut-mei=Apichart kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=BoonruamkaewPhetcharat en-aut-sei=Boonruamkaew en-aut-mei=Phetcharat kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KrobthongSucheewin en-aut-sei=Krobthong en-aut-mei=Sucheewin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=AonbangkhenChanat en-aut-sei=Aonbangkhen en-aut-mei=Chanat kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=UchiyamaJumpei en-aut-sei=Uchiyama en-aut-mei=Jumpei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YingchutrakulYodying en-aut-sei=Yingchutrakul en-aut-mei=Yodying kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=SongnakaNuttapon en-aut-sei=Songnaka en-aut-mei=Nuttapon kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=School of Pharmacy, Walailak University kn-affil= affil-num=2 en-affil=School of Pharmacy, Walailak University kn-affil= affil-num=3 en-affil=School of Pharmacy, Walailak University kn-affil= affil-num=4 en-affil=Center of Excellence in Natural Products Chemistry (CENP), Department of Chemistry, Faculty of Science, Chulalongkorn University kn-affil= affil-num=5 en-affil=Center of Excellence in Natural Products Chemistry (CENP), Department of Chemistry, Faculty of Science, Chulalongkorn University kn-affil= affil-num=6 en-affil=Department of Bacteriology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=7 en-affil=National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency kn-affil= affil-num=8 en-affil=School of Pharmacy, Walailak University kn-affil= en-keyword=anionic AMP kn-keyword=anionic AMP en-keyword=AMP kn-keyword=AMP en-keyword=antimicrobial peptide kn-keyword=antimicrobial peptide en-keyword=antimicrobial resistance kn-keyword=antimicrobial resistance en-keyword=FN33 kn-keyword=FN33 en-keyword=genome kn-keyword=genome en-keyword=molecular dynamics simulations kn-keyword=molecular dynamics simulations en-keyword=MRSA kn-keyword=MRSA en-keyword=Virgibacillus chiguensis kn-keyword=Virgibacillus chiguensis END start-ver=1.4 cd-journal=joma no-vol=13 cd-vols= no-issue=9 article-no= start-page=846 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240905 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Unveiling a New Antimicrobial Peptide with Efficacy against P. aeruginosa and K. pneumoniae from Mangrove-Derived Paenibacillus thiaminolyticus NNS5-6 and Genomic Analysis en-subtitle= kn-subtitle= en-abstract= kn-abstract=This study focused on the discovery of the antimicrobial peptide (AMP) derived from mangrove bacteria. The most promising isolate, NNS5-6, showed the closest taxonomic relation to Paenibacillus thiaminolyticus, with the highest similarity of 74.9%. The AMP produced by Paenibacillus thiaminolyticus NNS5-6 exhibited antibacterial activity against various Gram-negative pathogens, especially Pseudomonas aeruginosa and Klebsiella pneumoniae. The peptide sequence consisted of 13 amino acids and was elucidated as Val-Lys-Gly-Asp-Gly-Gly-Pro-Gly-Thr-Val-Tyr-Thr-Met. The AMP mainly exhibited random coil and antiparallel beta-sheet structures. The stability study indicated that this AMP was tolerant of various conditions, including proteolytic enzymes, pH (1.2?14), surfactants, and temperatures up to 40 ‹C for 12 h. The AMP demonstrated 4 ?g/mL of MIC and 4?8 ?g/mL of MBC against both pathogens. Time-kill kinetics showed that the AMP acted in a time- and concentration-dependent manner. A cell permeability assay and scanning electron microscopy revealed that the AMP exerted the mode of action by disrupting bacterial membranes. Additionally, nineteen biosynthetic gene clusters of secondary metabolites were identified in the genome. NNS5-6 was susceptible to various commonly used antibiotics supporting the primary safety requirement. The findings of this research could pave the way for new therapeutic approaches in combating antibiotic-resistant pathogens. en-copyright= kn-copyright= en-aut-name=SermkaewNamfa en-aut-sei=Sermkaew en-aut-mei=Namfa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AtipairinApichart en-aut-sei=Atipairin en-aut-mei=Apichart kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KrobthongSucheewin en-aut-sei=Krobthong en-aut-mei=Sucheewin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=AonbangkhenChanat en-aut-sei=Aonbangkhen en-aut-mei=Chanat kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YingchutrakulYodying en-aut-sei=Yingchutrakul en-aut-mei=Yodying kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=UchiyamaJumpei en-aut-sei=Uchiyama en-aut-mei=Jumpei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SongnakaNuttapon en-aut-sei=Songnaka en-aut-mei=Nuttapon kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=School of Pharmacy, Walailak University kn-affil= affil-num=2 en-affil=School of Pharmacy, Walailak University kn-affil= affil-num=3 en-affil=Center of Excellence in Natural Products Chemistry (CENP), Department of Chemistry, Faculty of Science, Chulalongkorn University kn-affil= affil-num=4 en-affil=Center of Excellence in Natural Products Chemistry (CENP), Department of Chemistry, Faculty of Science, Chulalongkorn University kn-affil= affil-num=5 en-affil=National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency kn-affil= affil-num=6 en-affil=Department of Bacteriology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=7 en-affil=School of Pharmacy, Walailak University kn-affil= en-keyword=antimicrobial peptide kn-keyword=antimicrobial peptide en-keyword=antimicrobial resistance kn-keyword=antimicrobial resistance en-keyword=bacterial genome kn-keyword=bacterial genome en-keyword=biosynthetic gene cluster kn-keyword=biosynthetic gene cluster en-keyword=Klebsiella pneumoniae kn-keyword=Klebsiella pneumoniae en-keyword=Mangrove kn-keyword=Mangrove en-keyword=mass spectrometry kn-keyword=mass spectrometry en-keyword=NNS5-6 kn-keyword=NNS5-6 en-keyword=Paenibacillus thiaminolyticus kn-keyword=Paenibacillus thiaminolyticus en-keyword=Pseudomonas aeruginosa kn-keyword=Pseudomonas aeruginosa END start-ver=1.4 cd-journal=joma no-vol=1869 cd-vols= no-issue=12 article-no= start-page=130860 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250913 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The F54L mutation of Thioredoxin shows protein instability and increased fluctuations of the catalytic center en-subtitle= kn-subtitle= en-abstract= kn-abstract=Thioredoxin is a ubiquitous redox protein that acts as an electron donor via its conserved dithiol motif (C32GPC35), catalyzing dithiol?disulfide exchange to regulate the redox state of target proteins. It supports antioxidant defense via peroxiredoxins, facilitates DNA synthesis by donating electrons to ribonucleotide reductase, and regulates redox-sensitive signaling pathways, including those controlling transcription and apoptosis. Neuronal degeneration and chronic kidney disease have been observed in Txn-F54L mutant rats; however, the details of why the Txn mutation causes these phenomena remain unknown. The present study aimed to elucidate the functional and structural changes caused by the F54L mutation. The Thioredoxin-F54L showed less insulin-reducing activity and more thermosensitivity to denaturation in the body temperature range compared to the wild type. The crystal structure revealed that F54 forms hydrophobic interactions with the surrounding hydrophobic amino acids. In addition, molecular dynamics simulation predicts increased fluctuations around the F54L mutation and a tendency for the distance between residues C32 and C35 at the catalytic center to be widened. The increased distance between residues C32 and C35 of the catalytic center may affect the reducing activity of the enzyme on the substrate. The finding that Thioredoxin-F54L is prone to denaturation at normal body temperature may reduce the normally functioning Thioredoxin. These molecular characteristics of Thioredoxin-F54L may be related to brain and kidney disease development in the Txn-F54L rats. en-copyright= kn-copyright= en-aut-name=BabaTakumi en-aut-sei=Baba en-aut-mei=Takumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=UenoGo en-aut-sei=Ueno en-aut-mei=Go kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OheChika en-aut-sei=Ohe en-aut-mei=Chika kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SajiShuku en-aut-sei=Saji en-aut-mei=Shuku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamamotoSachiko en-aut-sei=Yamamoto en-aut-mei=Sachiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=YamamotoMasaki en-aut-sei=Yamamoto en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NakagawaHiroshi en-aut-sei=Nakagawa en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=OkazakiNobuo en-aut-sei=Okazaki en-aut-mei=Nobuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=OuchidaMamoru en-aut-sei=Ouchida en-aut-mei=Mamoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=Kawasaki-OhmoriIori en-aut-sei=Kawasaki-Ohmori en-aut-mei=Iori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TakeshitaKohei en-aut-sei=Takeshita en-aut-mei=Kohei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Life Science Research Infrastructure Group, Advanced Photon Technology Division, RIKEN SPring-8 Center kn-affil= affil-num=2 en-affil=Life Science Research Infrastructure Group, Advanced Photon Technology Division, RIKEN SPring-8 Center kn-affil= affil-num=3 en-affil=Life Science Research Infrastructure Group, Advanced Photon Technology Division, RIKEN SPring-8 Center kn-affil= affil-num=4 en-affil=Structural Biology Division, Japan Synchrotron Radiation Research Institute kn-affil= affil-num=5 en-affil=Structural Biology Division, Japan Synchrotron Radiation Research Institute kn-affil= affil-num=6 en-affil=Life Science Research Infrastructure Group, Advanced Photon Technology Division, RIKEN SPring-8 Center kn-affil= affil-num=7 en-affil=Materials Sciences Research Center, Japan Atomic Energy Agency kn-affil= affil-num=8 en-affil=Neutron Science and Technology Center, Comprehensive Research Organization for Science and Society (CROSS) kn-affil= affil-num=9 en-affil=Department of Molecular Oncology, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=10 en-affil=Section of Developmental Physiology and Pathology, Faculty of Education, Okayama University kn-affil= affil-num=11 en-affil=Life Science Research Infrastructure Group, Advanced Photon Technology Division, RIKEN SPring-8 Center kn-affil= en-keyword=Txn kn-keyword=Txn en-keyword=Thioredoxin kn-keyword=Thioredoxin en-keyword=Protein instability kn-keyword=Protein instability en-keyword=Thermosensitivity kn-keyword=Thermosensitivity en-keyword=Crystal structure kn-keyword=Crystal structure en-keyword=Molecular dynamics simulation kn-keyword=Molecular dynamics simulation END start-ver=1.4 cd-journal=joma no-vol=89 cd-vols= no-issue=8 article-no= start-page=1217 end-page=1226 dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250527 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Microbial biotransformation of proteins into amino acids in unpolished Thai and polished Japanese rice varieties cultivated with distinct industrial strains of koji mold en-subtitle= kn-subtitle= en-abstract= kn-abstract=We previously reported the cultivation of industrial koji mold strains to produce unpolished Thai-colored rice kojis. These kojis, along with those made from unpolished Thai white rice and polished Japanese white rice, showed increased polyphenol content after cultivation, with the highest levels observed in unpolished Thai-colored rice kojis. In this study, an increase in both proteinogenic and non-proteinogenic amino acid contents, particularly ƒΑ-aminobutyric acid (GABA) content, was observed in both unpolished Thai and polished Japanese rice kojis, suggesting the ability of koji mold in the biotransformation of proteins. This increase was almost comparable even when using different rice varieties; in contrast, it varied depending on the koji mold strain used. The observed increase in both polyphenol and functional amino acid contents, especially GABA content, highlights the potential of unpolished Thai and polished Japanese rice kojis, particularly unpolished Thai-colored rice koji, as multifunctional materials, benefiting from polyphenol and amino acid functionalities. en-copyright= kn-copyright= en-aut-name=JitpakdeeJirayu en-aut-sei=Jitpakdee en-aut-mei=Jirayu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ItoKazunari en-aut-sei=Ito en-aut-mei=Kazunari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TaninoYuka en-aut-sei=Tanino en-aut-mei=Yuka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakeuchiHayato en-aut-sei=Takeuchi en-aut-mei=Hayato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamashitaHideyuki en-aut-sei=Yamashita en-aut-mei=Hideyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NakagawaTakuro en-aut-sei=Nakagawa en-aut-mei=Takuro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NitodaTeruhiko en-aut-sei=Nitoda en-aut-mei=Teruhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KanzakiHiroshi en-aut-sei=Kanzaki en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Graduate School of Environmental, Life, Natural Science and Technology, Okayama University kn-affil= affil-num=2 en-affil=Industrial Technology Center of Okayama Prefecture kn-affil= affil-num=3 en-affil=Industrial Technology Center of Okayama Prefecture kn-affil= affil-num=4 en-affil=Industrial Technology Center of Okayama Prefecture kn-affil= affil-num=5 en-affil=Higuchi Matsunosuke Shoten Co., Ltd. kn-affil= affil-num=6 en-affil=Higuchi Matsunosuke Shoten Co., Ltd. kn-affil= affil-num=7 en-affil=Graduate School of Environmental, Life, Natural Science and Technology, Okayama University kn-affil= affil-num=8 en-affil=Graduate School of Environmental, Life, Natural Science and Technology, Okayama University kn-affil= en-keyword=Amino acid kn-keyword=Amino acid en-keyword=GABA kn-keyword=GABA en-keyword=koji mold kn-keyword=koji mold en-keyword=rice koji kn-keyword=rice koji en-keyword=Thai-colored rice kn-keyword=Thai-colored rice END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page=2503029 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250601 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Polyglycerol]Grafted Graphene Oxide with pH]Responsive Charge]Convertible Surface to Dynamically Control the Nanobiointeractions for Enhanced in Vivo Tumor Internalization en-subtitle= kn-subtitle= en-abstract= kn-abstract=pH-responsive charge-convertible nanomaterials (NMs) ameliorate the treatment of cancer via simultaneously reducing nonspecific interactions during systemic circulation and improving targeted uptake within solid tumors. While promising, little is known about how the pH-responsiveness of charge-convertible NMs directs their interactions with biological systems, leading to compromised performance, including off-target retention and low specificity to tumor cells. In the present study, polyglycerol-grafted graphene oxide bearing amino groups (GOPGNH2) at different densities are reacted with dimethylmaleic anhydride (DMMA), a pH-responsive moiety, to generate a set of charge-convertible GOPGNH-DMMA variants. This permits the assessment of a quantitative correlation between the structure of GOPGNH-DMMA to their pH-responsiveness, their dynamic interactions with proteins and cells, as well as their in vivo biological fate. Through a systematic investigation, it is revealed that GOPGNH115-DMMA prepared from GOPGNH2 with higher amine density experienced fast charge conversion at pH 7.4 to induce non-specific interactions at early stages, whereas GOPGNH60-DMMA and GOPGNH30-DMMA prepared from lower amine density retarded off-target charge conversion to enhance tumor accumulation. Notably, GOPGNH60-DMMA is also associated with enough amounts of proteins under acidic conditions to promote in vivo tumor internalization. The findings will inform the design of pH-responsive NMs for enhanced treatment accuracy and efficacy. en-copyright= kn-copyright= en-aut-name=ZouYajuan en-aut-sei=Zou en-aut-mei=Yajuan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=BiancoAlberto en-aut-sei=Bianco en-aut-mei=Alberto kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NishinaYuta en-aut-sei=Nishina en-aut-mei=Yuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=2 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=3 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= en-keyword=charge conversion kn-keyword=charge conversion en-keyword=in vivo tumor internalization kn-keyword=in vivo tumor internalization en-keyword=non-specific interaction kn-keyword=non-specific interaction en-keyword=pH-responsiveness kn-keyword=pH-responsiveness en-keyword=polyglycerol-grafted graphene oxide kn-keyword=polyglycerol-grafted graphene oxide END start-ver=1.4 cd-journal=joma no-vol=653 cd-vols= no-issue= article-no= start-page=119205 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=202503 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Meteoritic and asteroidal amino acid heterogeneity: Implications for planetesimal alteration conditions and sample return missions en-subtitle= kn-subtitle= en-abstract= kn-abstract=Carbonaceous chondrites (CC) and asteroid return samples contain amino acids (AA), which are essential for an origin of life on the early Earth and can provide important information concerning planetesimal alteration processes. While many studies have investigated AA from CC, separate studies have often found differing abundances for the same meteorite. Accordingly, analytical bias, differing terrestrial contamination levels and intrinsic sample heterogeneity have been proposed as potential reasons. However, current analytical techniques allow for the analysis of several mg-sized samples and can thus enable an investigation of AA heterogeneity within single meteorite specimens. Here, such an analytical technique is applied to characterise the AA in triplicate aliquots of three CCs. The results indicate that CCs are heterogenous in terms of their AA at the mm-scale. Furthermore, the results help to further constrain the effects of planetesimal alteration on organic matter and the requirements of future sample return missions that aim to obtain organic-bearing extraterrestrial materials. en-copyright= kn-copyright= en-aut-name=PotiszilChristian en-aut-sei=Potiszil en-aut-mei=Christian kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OtaTsutomu en-aut-sei=Ota en-aut-mei=Tsutomu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamanakaMasahiro en-aut-sei=Yamanaka en-aut-mei=Masahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KobayashiKatsura en-aut-sei=Kobayashi en-aut-mei=Katsura kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TanakaRyoji en-aut-sei=Tanaka en-aut-mei=Ryoji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NakamuraEizo en-aut-sei=Nakamura en-aut-mei=Eizo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= affil-num=2 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= affil-num=3 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= affil-num=4 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= affil-num=5 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= affil-num=6 en-affil=Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University kn-affil= en-keyword=Carbonaceous chondrite kn-keyword=Carbonaceous chondrite en-keyword=Heterogeneity kn-keyword=Heterogeneity en-keyword=Planetesimal kn-keyword=Planetesimal en-keyword=Aqueous alteration kn-keyword=Aqueous alteration en-keyword=Amino acid and meteorite kn-keyword=Amino acid and meteorite END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250603 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Amino Acid Substitutions in Loop C of Arabidopsis PIP2 Aquaporins Alters the Permeability of CO2 en-subtitle= kn-subtitle= en-abstract= kn-abstract=The transport of CO2 across biomembranes in plant cells is essential for efficient photosynthesis. Some aquaporins capable of CO2 transport, referred to as eCOOporinsf, are postulated to play a crucial role in leaf CO2 diffusion. However, the structural basis of CO2 permeation through aquaporins remains largely unknown. Here, we show that amino acids in loop C are critical for the CO2 permeability of Arabidopsis thaliana PIP2 aquaporins. We found that swapping tyrosine and serine in loop C to histidine and phenylalanine, which differ between AtPIP2;1 and AtPIP2;3, altered CO2 permeability when examined in the Xenopus laevis oocyte heterologous expression system. AlphaFold2 modelling indicated that these substitution induced a conformational shift in the sidechain of arginine in the aromatic/arginine (ar/R) selectivity filter and in lysine at the extracellular mouth of the monomeric pore in PIP2 aquaporins. Our findings demonstrate that distal amino acid substitutions can trigger conformational changes of the ar/R filter in the monomeric pore, modulating CO2 permeability. Additionally, phylogenetic analysis suggested that aquaporins capable of dual water/CO2 permeability are ancestral to those that are water-selective and CO2-impermeable, and CO2-selective and water impermeable. en-copyright= kn-copyright= en-aut-name=TaniaShaila Shermin en-aut-sei=Tania en-aut-mei=Shaila Shermin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=UtsugiShigeko en-aut-sei=Utsugi en-aut-mei=Shigeko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TsuchiyaYoshiyuki en-aut-sei=Tsuchiya en-aut-mei=Yoshiyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SasanoShizuka en-aut-sei=Sasano en-aut-mei=Shizuka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KatsuharaMaki en-aut-sei=Katsuhara en-aut-mei=Maki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MoriIzumi C. en-aut-sei=Mori en-aut-mei=Izumi C. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=4 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=5 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=6 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=Arabidopsis thaliana kn-keyword=Arabidopsis thaliana en-keyword=CO2 transport kn-keyword=CO2 transport en-keyword=monomeric pore kn-keyword=monomeric pore en-keyword=PIP2 aquaporin kn-keyword=PIP2 aquaporin en-keyword=Xenopus laevis kn-keyword=Xenopus laevis END start-ver=1.4 cd-journal=joma no-vol=16 cd-vols= no-issue=26 article-no= start-page=12024 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=2025 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Collective motions in the primary coordination sphere: a critical functional framework for catalytic activity of the oxygen-evolving complex of photosystem II en-subtitle= kn-subtitle= en-abstract= kn-abstract=Photosynthetic water oxidation, vital for dioxygen production and light energy conversion, is catalyzed by the oxygen-evolving complex of photosystem II, where the inorganic Mn4CaO5 cluster acts as the catalytic core. In this study, we investigate the functional significance of collective motions of amino acid side chains within the primary coordination sphere of the Mn cluster, focusing on their role in modulating the energetic demands for catalytic transformations in the S3 state. We applied regularized canonical correlation analysis to quantitatively correlate the three-dimensional arrangement of coordinating atoms with catalytic driving forces computed via density functional theory. Our analysis reveals that distinct collective side chain motions profoundly influence the energetic requirements for structural reconfigurations of the Mn cluster, achieved through expansion and contraction of the ligand cavity while fine-tuning its geometry to stabilize key intermediates. Complementary predictions from a neural network-based machine learning model indicate that the coordination sphere exerts a variable energetic impact on the catalytic transformations of the Mn cluster, depending on the S-state environment. Integrated computational analyses suggest that the extended lifetime of the S3YZ? state, consistently observed after three flash illuminations, may result from slow, progressive protein dynamics that continuously reshape the energy landscape, thereby shifting the equilibrium positions of rapid, reversible chemical processes over time. Overall, our findings demonstrate that collective motions in the primary coordination sphere constitute an active, dynamic framework essential for the efficient execution of multi-electron catalysis under ambient conditions, while simultaneously achieving a high selectivity with irreversible nature required for effective 3O2 evolution. en-copyright= kn-copyright= en-aut-name=IsobeHiroshi en-aut-sei=Isobe en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SuzukiTakayoshi en-aut-sei=Suzuki en-aut-mei=Takayoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SugaMichihiro en-aut-sei=Suga en-aut-mei=Michihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ShenJian-Ren en-aut-sei=Shen en-aut-mei=Jian-Ren kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamaguchiKizashi en-aut-sei=Yamaguchi en-aut-mei=Kizashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil= affil-num=2 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=3 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=4 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=5 en-affil=Center for Quantum Information and Quantum Biology, Osaka University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=5 article-no= start-page=489 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250430 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Mutagenesis Targeting the S153 Residue Within the Transmembrane ƒΐ-Hairpin of Mosquito-Larvicidal Mpp46Ab Affects Its Toxicity and the Synergistic Toxicity with Cry4Aa en-subtitle= kn-subtitle= en-abstract= kn-abstract=We constructed a library of Mpp46Ab mutants, in which S153 within the transmembrane ƒΐ-hairpin was randomly replaced by other amino acids. Mutagenesis and subsequent primary screening yielded 10 different Mpp46Ab mutants in addition to the wild type. Remarkably, S153 was replaced with a more hydrophobic amino acid in most of the mutants, and the S153I mutant in particular exhibited significantly increased toxicity. Electrophysiologic analysis using artificial lipid bilayers revealed that the single-channel conductance and PK/PCl permeability ratio were significantly increased for S153I pores. This suggests that the formation of highly ion-permeable and highly cation-selective toxin pores increases the influx of cations and water into cells, thereby facilitating osmotic shock. In addition, the S153F, S153L, and S153I mutants exhibited significantly reduced synergistic toxicity with Cry4Aa. Electrophysiologic analysis showed that the S153F, S153L, and S153I mutants form toxin pores with a significantly reduced PK/PNa permeability ratio and a significantly increased PK/PCa permeability ratio compared to wild-type pores. Thus, our results suggest that pore formation is central to the insecticidal activity of Mpp46Ab and that the ion permeability of toxin pores is a potential indicator correlated with both toxicity and synergistic toxicity with other toxins. en-copyright= kn-copyright= en-aut-name=HayakawaTohru en-aut-sei=Hayakawa en-aut-mei=Tohru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamaokaSyun en-aut-sei=Yamaoka en-aut-mei=Syun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AsakuraMami en-aut-sei=Asakura en-aut-mei=Mami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HiranoMinako en-aut-sei=Hirano en-aut-mei=Minako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=IdeToru en-aut-sei=Ide en-aut-mei=Toru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= en-keyword=Bacillus thuringiensis kn-keyword=Bacillus thuringiensis en-keyword=mosquito-larvicidal proteins kn-keyword=mosquito-larvicidal proteins en-keyword=synergistic toxicity kn-keyword=synergistic toxicity en-keyword=Culex pipiens mosquito larvae kn-keyword=Culex pipiens mosquito larvae en-keyword=side-directed mutagenesis kn-keyword=side-directed mutagenesis en-keyword=electrophysiologic analysis kn-keyword=electrophysiologic analysis END start-ver=1.4 cd-journal=joma no-vol=227 cd-vols= no-issue= article-no= start-page=110168 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=202510 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The hidden cation-selective pore in ion-conducting aquaporin OsPIP2;4 from rice en-subtitle= kn-subtitle= en-abstract= kn-abstract=Ion-conducting aquaporins (icAQPs) transport ions as well as water. Although the molecular mechanism of how AQPs establish selective permeability for water molecules is well understood, the ion-transporting mechanism in icAQPs has not yet been fully elucidated. In this study, we investigated the molecular mechanism of cation transport in OsPIP2;4, an icAQP in rice, by homology modeling and the electrophysiological analysis using Xenopus laevis oocytes. Water and ion transport assays using OsPIP2;4 T227M and G278K mutants strongly suggested that water- and cation-transporting pathways are independent of each other. Data from amino acid substitutions V54I and A143G in OsPIP2;4 led to the identification of a novel hidden pathway for cation transport located on the side surfaces of the tetramer channel, where two protomers are in contact, which is distinct from conventional monomeric pores and the tetrameric central pore in AQPs. Moreover, the present results provide the possibility that this hypothetical hidden pore also functions in the barley icAQP HvPIP2;8. The overall structure of this novel pathway appears to differ from the structure of general cation channels. However, the arrangement of hydrophilic amino acids at the entrance of the pathway of OsPIP2;4 was found to be comparable to that of some cation channels, which implies that the molecular mechanism of dehydration of hydrated ions might resemble that of the channels. Although direct structural evidence is needed to confirm the proposed pathway, the present study can be a stepping stone toward unraveling the mechanism of dual water and ion transport through icAQPs in plants. en-copyright= kn-copyright= en-aut-name=OnoShuntaro en-aut-sei=Ono en-aut-mei=Shuntaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TranSen Thi Huong en-aut-sei=Tran en-aut-mei=Sen Thi Huong kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SaitohYasunori en-aut-sei=Saitoh en-aut-mei=Yasunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=UtsugiShigeko en-aut-sei=Utsugi en-aut-mei=Shigeko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=HorieTomoaki en-aut-sei=Horie en-aut-mei=Tomoaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KatsuharaMaki en-aut-sei=Katsuhara en-aut-mei=Maki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=4 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=5 en-affil=Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University kn-affil= affil-num=6 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=Rice kn-keyword=Rice en-keyword=Barley kn-keyword=Barley en-keyword=Ion transport kn-keyword=Ion transport en-keyword=Ion-conducting aquaporin (icAQP) kn-keyword=Ion-conducting aquaporin (icAQP) en-keyword=Plasma membrane intrinsic protein (PIP) kn-keyword=Plasma membrane intrinsic protein (PIP) END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250526 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Lytic Transglycosylase Deficiency Increases Susceptibility to ƒΐ-lactam Antibiotics But Reduces Susceptibility to Vancomycin in Escherichia coli en-subtitle= kn-subtitle= en-abstract= kn-abstract=In Staphylococcus aureus, a gram-positive pathogen, vancomycin-resistant strains become susceptible to ƒΐ-lactam antibiotics, referred to as the gseesaw effect.h However, in gram-negative bacteria, the phenomenon is less clear. Here, we analyzed the gene-knockout effects of eight lytic transglycosylases (slt, mltA, mltB, mltC, mltD, mltE, mltF, mltG) on antibiotic sensitivity in Escherichia coli. Knockout of both slt and mltG increased sensitivity to ƒΐ-lactam antibiotics and reduced sensitivity to vancomycin. The ƒΐ-lactam antibiotic sensitivity and vancomycin resistance of the slt-knockout mutant were abolished by the introduction of the wild-type slt gene but remained unchanged by the introduction of the mutant slt gene encoding an amino acid substitution variant of the transglycosylase catalytic centre. The double-knockout strain for slt and mltB was more sensitive to ampicillin and more resistant to vancomycin than each single-knockout strain. The double-knockout strain for slt and mltG was more sensitive to ampicillin and more resistant to vancomycin than each single-knockout strain. These results suggest that loss of lytic transglycosylase activity causes ƒΐ-lactam antibiotic sensitivity and vancomycin resistance in E. coli. en-copyright= kn-copyright= en-aut-name=KimuraTakahiko en-aut-sei=Kimura en-aut-mei=Takahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IshikawaKazuya en-aut-sei=Ishikawa en-aut-mei=Kazuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NakagawaRyosuke en-aut-sei=Nakagawa en-aut-mei=Ryosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=FurutaKazuyuki en-aut-sei=Furuta en-aut-mei=Kazuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KaitoChikara en-aut-sei=Kaito en-aut-mei=Chikara kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Laboratory of Molecular Biology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Laboratory of Molecular Biology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Laboratory of Molecular Biology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Laboratory of Molecular Biology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Laboratory of Molecular Biology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=Escherichia coli kn-keyword=Escherichia coli en-keyword=lytic transglycosylase kn-keyword=lytic transglycosylase en-keyword=seesaw effect kn-keyword=seesaw effect en-keyword=vancomycin kn-keyword=vancomycin en-keyword=ƒΐ]lactam antibiotics kn-keyword=ƒΐ]lactam antibiotics END start-ver=1.4 cd-journal=joma no-vol=16 cd-vols= no-issue=1 article-no= start-page=2323 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250308 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A mini-hairpin shaped nascent peptide blocks translation termination by a distinct mechanism en-subtitle= kn-subtitle= en-abstract= kn-abstract=Protein synthesis by ribosomes produces functional proteins but also serves diverse regulatory functions, which depend on the coding amino acid sequences. Certain nascent peptides interact with the ribosome exit tunnel to arrest translation and modulate themselves or the expression of downstream genes. However, a comprehensive understanding of the mechanisms of such ribosome stalling and its regulation remains elusive. In this study, we systematically screen for unidentified ribosome arrest peptides through phenotypic evaluation, proteomics, and mass spectrometry analyses, leading to the discovery of the arrest peptides PepNL and NanCL in E. coli. Our cryo-EM study on PepNL reveals a distinct arrest mechanism, in which the N-terminus of PepNL folds back towards the tunnel entrance to prevent the catalytic GGQ motif of the release factor from accessing the peptidyl transferase center, causing translation arrest at the UGA stop codon. Furthermore, unlike sensory arrest peptides that require an arrest inducer, PepNL uses tryptophan as an arrest inhibitor, where Trp-tRNATrp reads through the stop codon. Our findings illuminate the mechanism and regulatory framework of nascent peptide-induced translation arrest, paving the way for exploring regulatory nascent peptides. en-copyright= kn-copyright= en-aut-name=AndoYushin en-aut-sei=Ando en-aut-mei=Yushin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KoboAkinao en-aut-sei=Kobo en-aut-mei=Akinao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NiwaTatsuya en-aut-sei=Niwa en-aut-mei=Tatsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamakawaAyako en-aut-sei=Yamakawa en-aut-mei=Ayako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KonomaSuzuna en-aut-sei=Konoma en-aut-mei=Suzuna kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KobayashiYuki en-aut-sei=Kobayashi en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NurekiOsamu en-aut-sei=Nureki en-aut-mei=Osamu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TaguchiHideki en-aut-sei=Taguchi en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=ItohYuzuru en-aut-sei=Itoh en-aut-mei=Yuzuru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=ChadaniYuhei en-aut-sei=Chadani en-aut-mei=Yuhei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil=Department of Biological Sciences, Graduate School of Science, The University of Tokyo kn-affil= affil-num=2 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=3 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=4 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=5 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=6 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=7 en-affil=Department of Biological Sciences, Graduate School of Science, The University of Tokyo kn-affil= affil-num=8 en-affil=School of Life Science and Technology, Institute of Science Tokyo kn-affil= affil-num=9 en-affil=Department of Biological Sciences, Graduate School of Science, The University of Tokyo kn-affil= affil-num=10 en-affil=Faculty of Environmental, Life, Natural Science and Technology, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=301 cd-vols= no-issue=4 article-no= start-page=108334 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=202504 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Roles of basic amino acid residues in substrate binding and transport of the light-driven anion pump Synechocystis halorhodopsin (SyHR) en-subtitle= kn-subtitle= en-abstract= kn-abstract=Microbial rhodopsins are photoreceptive seventransmembrane a-helical proteins, many of which function as ion transporters, primarily for small monovalent ions such as Na+, K+, Cl-, Br-, and I-. Synechocystis halorhodopsin (SyHR), identified from the cyanobacterium Synechocystis sp. PCC 7509, uniquely transports the polyatomic divalent SO42- inward, in addition to monovalent anions (Cl- and Br-). In this study, we conducted alanine-scanning mutagenesis on twelve basic amino acid residues to investigate the anion transport mechanism of SyHR. We quantitatively evaluated the Cl-and SO42- transport activities of the WT SyHR and its mutants. The results showed a strong correlation between the Cl-and SO42- transport activities among them (R = 0.94), suggesting a shared pathway for both anions. Notably, the R71A mutation selectively abolished SO42- transport activity while maintaining Cl- transport, whereas the H167A mutation significantly impaired both Cl-and SO42- transport. Furthermore, spectroscopic analysis revealed that the R71A mutant lost its ability to bind SO42- due to the absence of a positive charge, while the H167A mutant failed to accumulate the O intermediate during the photoreaction cycle (photocycle) due to reduced hydrophilicity. Additionally, computational analysis revealed the SO42- binding modes and clarified the roles of residues involved in its binding around the retinal chromophore. Based on these findings and previous structural information, we propose that the positive charge and hydrophilicity of Arg71 and His167 are crucial for the formation of the characteristic initial and transient anion-binding site of SyHR, enabling its unique ability to bind and transport both Cl-and SO42-. en-copyright= kn-copyright= en-aut-name=NakamaMasaki en-aut-sei=Nakama en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NojiTomoyasu en-aut-sei=Noji en-aut-mei=Tomoyasu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KojimaKeiichi en-aut-sei=Kojima en-aut-mei=Keiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YoshizawaSusumu en-aut-sei=Yoshizawa en-aut-mei=Susumu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=IshikitaHiroshi en-aut-sei=Ishikita en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SudoYuki en-aut-sei=Sudo en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Applied Chemistry, The University of Tokyo kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Atmosphere and Ocean Research Institute, The University of Tokyo kn-affil= affil-num=5 en-affil=Department of Applied Chemistry, The University of Tokyo kn-affil= affil-num=6 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=microbial rhodopsin kn-keyword=microbial rhodopsin en-keyword=anion transport kn-keyword=anion transport en-keyword=retinal kn-keyword=retinal en-keyword=membrane protein kn-keyword=membrane protein en-keyword=photobiology kn-keyword=photobiology END start-ver=1.4 cd-journal=joma no-vol=114 cd-vols= no-issue= article-no= start-page=21 end-page=25 dt-received= dt-revised= dt-accepted= dt-pub-year=2025 dt-pub=20250201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Elucidation of plant-bacterial pathogen interactions for the control of bacterial blight on cruciferous crops kn-title=ƒAƒuƒ‰ƒi‰ΘA•¨•”ΑΧ‹Ϋ•a‚Μ–hœ‚ΙŒό‚―‚½A•¨|•aŒ΄Χ‹Ϋ‚Μ‘ŠŒέμ—p‚Μ‰π–Ύ en-subtitle= kn-subtitle= en-abstract= kn-abstract=@Pseudomonas cannabina pv. alisalensis (Pcal), the causative agent of bacterial blight on cruciferous crops, is an economically important pathogen worldwide. We have conducted several studies on the interactions between plants and pathogenic bacteria to develop effective control strategies for this disease. Using forward and reverse genetics, we identified several virulence factors, including the type III secretion system, membrane transporters, transcriptional factors, and amino acid metabolism. Additionally, we emphasized the role of coronatine, a toxin produced by Pcal, which promotes stomatal reopening and suppresses salicylic acid accumulation in plants. We also examined plant defense mechanisms activated by one of the plant defense activators, acibenzolar-S-methyl (ASM). ASM enhanced stomatal-based defense, resulting in reduction of bacterial entry and disease development. Moreover, we explored innovative control strategies for bacterial disease and demonstrated that amino acids and cellulose nanofiber are efficient and environmentally friendly control strategies. These studies advance our understanding of plant-pathogen dynamics and offer promising, sustainable approaches for managing bacterial blight disease in cruciferous crops. en-copyright= kn-copyright= en-aut-name=SakataNanami en-aut-sei=Sakata en-aut-mei=Nanami kn-aut-name=β“cŽ΅ŠC kn-aut-sei=β“c kn-aut-mei=Ž΅ŠC aut-affil-num=1 ORCID= affil-num=1 en-affil=Course of Applied Plant Science kn-affil=‰ž—pA•¨‰ΘŠwƒR[ƒX en-keyword=Plant pathogenic bacteria kn-keyword=Plant pathogenic bacteria en-keyword=Pseudomonas kn-keyword=Pseudomonas en-keyword=Cruciferous kn-keyword=Cruciferous en-keyword=Plant protection kn-keyword=Plant protection en-keyword=Stomata kn-keyword=Stomata END start-ver=1.4 cd-journal=joma no-vol=39 cd-vols= no-issue=1 article-no= start-page=426 end-page=432 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20241231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effect of Oral Nutritional Supplements Composed of High Protein on Body Weight Loss After Gastrectomy en-subtitle= kn-subtitle= en-abstract= kn-abstract=Background/Aim: Body weight loss (BWL) after gastrectomy for gastric cancer (GC) decreases postoperative quality of life and survival in patients with GC. This study aimed to evaluate the effect of oral nutritional supplements composed of high protein on BWL in the early period following gastrectomy. Patients and Methods: Pre- and postoperative body weight and skeletal muscle mass were measured using bioelectrical impedance analysis in patients undergoing radical gastrectomy for GC and analyzed retrospectively. Patients received either a regular diet (control group, n=43) or 250 ml (320 kcal) per day of a high-protein oral nutritional supplement (ONS) (22 g protein) in addition to their regular diet (ONS group, n=40) for four weeks after gastrectomy. The actual daily intake of ONS was recorded by patients themselves. The BWL and skeletal muscle loss (SML) at one month after surgery were compared between the two groups. Results: BWL and SML at one month after surgery were similar between the two groups. In the ONS group, patients were divided into two subgroups (ONS-H and ONS-L) according to whether their ONS intake amount was above or below the average value of 216 kcal. The ONS-H group (ONS intake ?216 kcal) showed significantly lower BWL compared to the control group (?4.6}2.6% vs. ?6.2}2.5%; p=0.03). Moreover, the ONS group showed significantly lower BWL at one month after surgery than the control group in cases of total or proximal gastrectomy (?5.9}3.0% vs. ?7.8}1.9%; p=0.04), although no significant difference was observed between the two groups in distal gastrectomy. The hematological nutritional parameters were similar between the two groups. Conclusion: The administration of ONS composed of high protein for four weeks after gastrectomy did not improve BWL at one month after gastrectomy. However, adequate amount of ONS intake and ONS intake after total or proximal gastrectomy might improve BWL. en-copyright= kn-copyright= en-aut-name=KIKUCHISATORU en-aut-sei=KIKUCHI en-aut-mei=SATORU kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TAKATANOBUO en-aut-sei=TAKATA en-aut-mei=NOBUO kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KAKIUCHIYOSHIHIKO en-aut-sei=KAKIUCHI en-aut-mei=YOSHIHIKO kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KURODASHINJI en-aut-sei=KURODA en-aut-mei=SHINJI kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KASHIMAHAJIME en-aut-sei=KASHIMA en-aut-mei=HAJIME kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TANABESHUNSUKE en-aut-sei=TANABE en-aut-mei=SHUNSUKE kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NOMAKAZUHIRO en-aut-sei=NOMA en-aut-mei=KAZUHIRO kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TAKAHASHIAYAKO en-aut-sei=TAKAHASHI en-aut-mei=AYAKO kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KAGAWASHUNSUKE en-aut-sei=KAGAWA en-aut-mei=SHUNSUKE kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=FUJIWARATOSHIYOSHI en-aut-sei=FUJIWARA en-aut-mei=TOSHIYOSHI kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Gastroenterological Surgery, Kochi Health Sciences Center kn-affil= affil-num=3 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Clinical Nutrition, Okayama University Hospital kn-affil= affil-num=9 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=10 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=Amino acid kn-keyword=Amino acid en-keyword=gastrectomy kn-keyword=gastrectomy en-keyword=body weight loss kn-keyword=body weight loss en-keyword=nutritional intervention kn-keyword=nutritional intervention en-keyword=oral nutritional supplements kn-keyword=oral nutritional supplements END start-ver=1.4 cd-journal=joma no-vol=9 cd-vols= no-issue=50 article-no= start-page=50041 end-page=50048 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20241205 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Conformational Flexibility of D1-Glu189: A Crucial Determinant in Substrate Water Selection, Positioning, and Stabilization within the Oxygen-Evolving Complex of Photosystem II en-subtitle= kn-subtitle= en-abstract= kn-abstract=Photosynthetic water oxidation is a vital process responsible for producing dioxygen and supplying the energy necessary to sustain life on Earth. This fundamental reaction is catalyzed by the oxygen-evolving complex (OEC) of photosystem II, which houses the Mn4CaO5 cluster as its catalytic core. In this study, we specifically focus on the D1-Glu189 amino acid residue, which serves as a direct ligand to the Mn4CaO5 cluster. Our primary goal is to explore, using density functional theory (DFT), how the conformational flexibility of the D1-Glu189 side chain influences crucial catalytic processes, particularly the selection, positioning, and stabilization of a substrate water molecule within the OEC. Our investigation is based on a hypothesis put forth by Li et al. (Nature, 2024, 626, 670), which suggests that during the transition from the S2 to S3 state, a specific water molecule temporarily coordinating with the Ca ion, referred to as O6*, may exist as a hydroxide ion (OH-). Our results demonstrate a key mechanism by which the detachment of the D1-Glu189 carboxylate group from its coordination with the Ca ion allows the creation of a specialized microenvironment within the OEC that enables the selective attraction of O6* in its deprotonated form (OH-) and stabilizes it at the catalytic metal (MnD) site. Our findings indicate that D1-Glu189 is not only a structural ligand for the Ca ion but may also play an active and dynamic role in the catalytic process, positioning O6* optimally for its subsequent participation in the oxidation sequence during the water-splitting cycle. en-copyright= kn-copyright= en-aut-name=IsobeHiroshi en-aut-sei=Isobe en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SuzukiTakayoshi en-aut-sei=Suzuki en-aut-mei=Takayoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SugaMichihiro en-aut-sei=Suga en-aut-mei=Michihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ShenJian-Ren en-aut-sei=Shen en-aut-mei=Jian-Ren kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamaguchiKizashi en-aut-sei=Yamaguchi en-aut-mei=Kizashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=2 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=3 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=4 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=5 en-affil=Center for Quantum Information and Quantum Biology, Osaka University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=19 cd-vols= no-issue=10 article-no= start-page=e0310962 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20241023 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Examination of yield, bacteriolytic activity and cold storage of linker deletion mutants based on endolysin S6_ORF93 derived from Staphylococcus giant bacteriophage S6 en-subtitle= kn-subtitle= en-abstract= kn-abstract=Methicillin-resistant Staphylococcus spp. present challenges in clinical and veterinary settings because effective antimicrobial agents are limited. Phage-encoded peptidoglycan-degrading enzyme, endolysin, is expected to be a novel antimicrobial agent. The enzymatic activity has recently been shown to be influenced by the linker between functional domains in the enzyme. S6_ORF93 (ORF93) is one of the endolysins derived from previously isolated Staphylococcus giant phage S6. The ORF93 was speculated to have a catalytic and peptidoglycan-binding domain with a long linker. In this study, we examined the influence of linker shortening on the characteristics of ORF93. We produce wild-type ORF93 and the linker deletion mutants using an Escherichia coli expression system. These mutants were designated as ORF93-Delta 05, ORF93-Delta 10, ORF93-Delta 15, and ORF93-Delta 20, from which 5, 10, 15, and 20 amino acids were removed from the linker, respectively. Except for the ORF93-Delta 20, ORF93 and its mutants were expressed as soluble proteins. Moreover, ORF93-Delta 15 showed the highest yield and bacteriolytic activity, while the antimicrobial spectrum was homologous. The cold storage experiment showed a slight effect by the linker deletion. According to our results and other studies, linker investigations are crucial in endolysin development. en-copyright= kn-copyright= en-aut-name=MunetomoSosuke en-aut-sei=Munetomo en-aut-mei=Sosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=UchiyamaJumpei en-aut-sei=Uchiyama en-aut-mei=Jumpei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=Takemura-UchiyamaIyo en-aut-sei=Takemura-Uchiyama en-aut-mei=Iyo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=WanganuttaraThamonwan en-aut-sei=Wanganuttara en-aut-mei=Thamonwan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 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=5 ORCID= en-aut-name=TsukuiToshihiro en-aut-sei=Tsukui en-aut-mei=Toshihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=HagiyaHideharu en-aut-sei=Hagiya en-aut-mei=Hideharu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KanamaruShuji en-aut-sei=Kanamaru en-aut-mei=Shuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KandaHideyuki en-aut-sei=Kanda en-aut-mei=Hideyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 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=10 ORCID= affil-num=1 en-affil=Department of Public Health, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Bacteriology, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Department of Bacteriology, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Department of Bacteriology, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Department of Bacteriology, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=6 en-affil=Nippon Zenyaku Kogyo Co. Ltd. kn-affil= affil-num=7 en-affil=Department of Infectious Diseases, Okayama University Hospital kn-affil= affil-num=8 en-affil=School of Life Science and Technology, Tokyo Institute of Technology kn-affil= affil-num=9 en-affil=Department of Public Health, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=10 en-affil=Department of Bacteriology, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=78 cd-vols= no-issue=5 article-no= start-page=387 end-page=399 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=202410 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effect of Radon Inhalation on Murine Brain Proteins: Investigation Using Proteomic and Multivariate Analyses en-subtitle= kn-subtitle= en-abstract= kn-abstract=Radon is a known risk factor for lung cancer; however, it can be used beneficially, such as in radon therapy. We have previously reported the enhancement of antioxidant effects associated with trace amounts of oxidative stress as one of the positive biological effects of radon inhalation. However, the biological effects of radon inhalation are incompletely understood, and more detailed and comprehensive studies are required. Although several studies have used proteomics to investigate the effects of radon inhalation on body proteins, none has focused on brain proteins. In this study, we evaluated the expression status of proteins in murine brains using proteomic and multivariate analyses to identify those whose expressions changed following two days of radon inhalation at a concentration of 1,500 Bq/m3. We found associations of radon inhalation with the expressions of seven proteins related to neurotransmission and heat shock. These proteins may be proposed as biomarkers indicative of radon inhalation. Although further studies are required to obtain the detailed biological significance of these protein alterations, this study contributes to the elucidation of the biological effects of radon inhalation as a low-dose radiation. en-copyright= kn-copyright= en-aut-name=NaoeShota en-aut-sei=Naoe en-aut-mei=Shota kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TanakaAyumi en-aut-sei=Tanaka en-aut-mei=Ayumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KanzakiNorie en-aut-sei=Kanzaki en-aut-mei=Norie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakenakaReiju en-aut-sei=Takenaka en-aut-mei=Reiju kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SakodaAkihiro en-aut-sei=Sakoda en-aut-mei=Akihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MiyajiTakaaki en-aut-sei=Miyaji en-aut-mei=Takaaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YamaokaKiyonori en-aut-sei=Yamaoka en-aut-mei=Kiyonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KataokaTakahiro en-aut-sei=Kataoka en-aut-mei=Takahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Graduate School of Health Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Health Sciences, Okayama University kn-affil= affil-num=3 en-affil=Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency kn-affil= affil-num=4 en-affil=Graduate School of Health Sciences, Okayama University kn-affil= affil-num=5 en-affil=Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency kn-affil= affil-num=6 en-affil=Advanced Science Research Center, Okayama University kn-affil= affil-num=7 en-affil=Faculty of Health Sciences, Okayama University kn-affil= affil-num=8 en-affil=Faculty of Health Sciences, Okayama University kn-affil= en-keyword=radon inhalation kn-keyword=radon inhalation en-keyword=proteomics kn-keyword=proteomics en-keyword=multivariate analysis kn-keyword=multivariate analysis en-keyword=brain kn-keyword=brain en-keyword=oxidative stress kn-keyword=oxidative stress END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue=9 article-no= start-page=1781 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240828 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A Novel C-Terminal Truncated Bacteriocin Found by Comparison between Leuconostoc mesenteroides 406 and 213M0 Isolated from Mongolian Traditional Fermented Milk, Airag en-subtitle= kn-subtitle= en-abstract= kn-abstract=Bacteriocins produced by lactic acid bacteria are known to be useful tools for food biopreservation and fermentation control. Leuconostoc mesenteroides subsp. mesenteroides 406 and 213M0 isolated from different samples of Mongolian traditional fermented milk, airag, had been reported to produce listericidal bacteriocin-like inhibitory substances with similar but slightly different properties. In this study, the antibacterial properties and the related gene sequences of both strains were compared, and then their bacteriocins were purified and identified. Strain 406 was superior to strain 213M0 in cell growth and antibacterial activity against many strains. However, the activity of 213M0 was stronger than that of 406 against a few strains. DNA sequencing revealed two and three plasmids in 406 and 213M0, respectively, and each one of them harbored an almost identical mesentericin Y105-B105 gene cluster. Removal of these plasmids resulted in a complete loss of activity, indicating that the antibacterial activity of both strains was generated by bacteriocins encoded on the plasmids. Mesentericins Y105 and B105 were purified from both cultures, and another novel bacteriocin, named mesentericin M, was identified from the 213M0 culture only. Its structural gene was coded on a 213M0 plasmid and, surprisingly, its C-terminal three amino acid residues were post-translationally cleaved. To our knowledge, this is the first report of a C-terminal truncated bacteriocin. In conclusion, the novel bacteriocin should be mainly responsible for the difference in antibacterial properties between the two strains. en-copyright= kn-copyright= en-aut-name=HasiqimugeChihiro en-aut-sei=Hasiqimuge en-aut-mei=Chihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HanoChihiro en-aut-sei=Hano en-aut-mei=Chihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=ArakawaKensuke en-aut-sei=Arakawa en-aut-mei=Kensuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YoshidaSaki en-aut-sei=Yoshida en-aut-mei=Saki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ZhaoJunliang en-aut-sei=Zhao en-aut-mei=Junliang kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TohHidehiro en-aut-sei=Toh en-aut-mei=Hidehiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MoritaHidetoshi en-aut-sei=Morita en-aut-mei=Hidetoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MiyamotoTaku en-aut-sei=Miyamoto en-aut-mei=Taku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=6 en-affil=Advanced Genomics Center, National Institute of Genetics kn-affil= affil-num=7 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=8 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword=Leuconostoc mesenteroides kn-keyword=Leuconostoc mesenteroides en-keyword=antimicrobial peptide kn-keyword=antimicrobial peptide en-keyword=bacteriocin kn-keyword=bacteriocin en-keyword=Listeria monocytogenes kn-keyword=Listeria monocytogenes en-keyword=fermented milk kn-keyword=fermented milk en-keyword=biopreservation kn-keyword=biopreservation en-keyword=fermentation control kn-keyword=fermentation control en-keyword=post-translational modification kn-keyword=post-translational modification en-keyword=C-terminal cleavage kn-keyword=C-terminal cleavage END start-ver=1.4 cd-journal=joma no-vol=137 cd-vols= no-issue=9 article-no= start-page=212 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240831 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Mutations in starch BRANCHING ENZYME 2a suppress the traits caused by the loss of ISOAMYLASE1 in barley en-subtitle= kn-subtitle= en-abstract= kn-abstract=The genetic interactions among starch biosynthesis genes can be exploited to alter starch properties, but they remain poorly understood due to the various combinations of mutations to be tested. Here, we isolated two novel barley mutants defective in starch BRANCHING ENZYME 2a (hvbe2a-1 and hvbe2a-2) based on the starch granule (SG) morphology. Both hvbe2a mutants showed elongated SGs in the endosperm and increased resistant starch content. hvbe2a-1 had a base change in HvBE2a gene, substituting the amino acid essential for its enzyme activity, while hvbe2a-2 is completely missing HvBE2a due to a chromosomal deletion. Further genetic crosses with barley isoamylase1 mutants (hvisa1) revealed that both hvbe2a mutations could suppress defects in endosperm caused by hvisa1, such as reduction in starch, increase in phytoglycogen, and changes in the glucan chain length distribution. Remarkably, hvbe2a mutations also transformed the endosperm SG morphology from the compound SG caused by hvisa1 to bimodal simple SGs, resembling that of wild-type barley. The suppressive impact was in competition with floury endosperm 6 mutation (hvflo6), which could enhance the phenotype of hvisa1 in the endosperm. In contrast, the compound SG formation induced by the hvflo6 hvisa1 mutation in pollen was not suppressed by hvbe2a mutations. Our findings provide new insights into genetic interactions in the starch biosynthetic pathway, demonstrating how specific genetic alterations can influence starch properties and SG morphology, with potential applications in cereal breeding for desired starch properties. en-copyright= kn-copyright= en-aut-name=MatsushimaRyo en-aut-sei=Matsushima en-aut-mei=Ryo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HisanoHiroshi en-aut-sei=Hisano en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KimJune-Sik en-aut-sei=Kim en-aut-mei=June-Sik kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=McNellyRose en-aut-sei=McNelly en-aut-mei=Rose kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OitomeNaoko F. en-aut-sei=Oitome en-aut-mei=Naoko F. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SeungDavid en-aut-sei=Seung en-aut-mei=David kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=FujitaNaoko en-aut-sei=Fujita en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=SatoKazuhiro en-aut-sei=Sato en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=4 en-affil=John Innes Centre, Norwich Research Park kn-affil= affil-num=5 en-affil=Department of Biological Production, Akita Prefectural University kn-affil= affil-num=6 en-affil=John Innes Centre, Norwich Research Park kn-affil= affil-num=7 en-affil=Department of Biological Production, Akita Prefectural University kn-affil= affil-num=8 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=25 cd-vols= no-issue=15 article-no= start-page=8370 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240731 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Increased Oxidative Stress and Decreased Citrulline in Blood Associated with Severe Novel Coronavirus Pneumonia in Adult Patients en-subtitle= kn-subtitle= en-abstract= kn-abstract=This study investigated the correlation between oxidative stress and blood amino acids associated with nitric oxide metabolism in adult patients with coronavirus disease (COVID-19) pneumonia. Clinical data and serum samples were prospectively collected from 100 adult patients hospitalized for COVID-19 between July 2020 and August 2021. Patients with COVID-19 were categorized into three groups for analysis based on lung infiltrates, oxygen inhalation upon admission, and the initiation of oxygen therapy after admission. Blood data, oxidative stress-related biomarkers, and serum amino acid levels upon admission were compared in these groups. Patients with lung infiltrations requiring oxygen therapy upon admission or starting oxygen post-admission exhibited higher serum levels of hydroperoxides and lower levels of citrulline compared to the control group. No remarkable differences were observed in nitrite/nitrate, asymmetric dimethylarginine, and arginine levels. Serum citrulline levels correlated significantly with serum lactate dehydrogenase and C-reactive protein levels. A significant negative correlation was found between serum levels of citrulline and hydroperoxides. Levels of hydroperoxides decreased, and citrulline levels increased during the recovery period compared to admission. Patients with COVID-19 with extensive pneumonia or poor oxygenation showed increased oxidative stress and reduced citrulline levels in the blood compared to those with fewer pulmonary complications. These findings suggest that combined oxidative stress and abnormal citrulline metabolism may play a role in the pathogenesis of COVID-19 pneumonia. en-copyright= kn-copyright= en-aut-name=TsugeMitsuru en-aut-sei=Tsuge en-aut-mei=Mitsuru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IchiharaEiki en-aut-sei=Ichihara en-aut-mei=Eiki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HasegawaKou en-aut-sei=Hasegawa en-aut-mei=Kou kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KudoKenichiro en-aut-sei=Kudo en-aut-mei=Kenichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TanimotoYasushi en-aut-sei=Tanimoto en-aut-mei=Yasushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NousoKazuhiro en-aut-sei=Nouso en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=OdaNaohiro en-aut-sei=Oda en-aut-mei=Naohiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MitsumuneSho en-aut-sei=Mitsumune en-aut-mei=Sho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KimuraGoro en-aut-sei=Kimura en-aut-mei=Goro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=YamadaHaruto en-aut-sei=Yamada en-aut-mei=Haruto kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TakataIchiro en-aut-sei=Takata en-aut-mei=Ichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=MitsuhashiToshiharu en-aut-sei=Mitsuhashi en-aut-mei=Toshiharu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=TaniguchiAkihiko en-aut-sei=Taniguchi en-aut-mei=Akihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=TsukaharaKohei en-aut-sei=Tsukahara en-aut-mei=Kohei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=AokageToshiyuki en-aut-sei=Aokage en-aut-mei=Toshiyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= en-aut-name=HagiyaHideharu en-aut-sei=Hagiya en-aut-mei=Hideharu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=16 ORCID= en-aut-name=ToyookaShinichi en-aut-sei=Toyooka en-aut-mei=Shinichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=17 ORCID= en-aut-name=TsukaharaHirokazu en-aut-sei=Tsukahara en-aut-mei=Hirokazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=18 ORCID= en-aut-name=MaedaYoshinobu en-aut-sei=Maeda en-aut-mei=Yoshinobu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=19 ORCID= affil-num=1 en-affil=Department of Pediatrics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= affil-num=3 en-affil=Department of General Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Department of Respiratory Medicine, National Hospital Organization Minami-Okayama Medical Center kn-affil= affil-num=5 en-affil=Department of Allergy and Respiratory Medicine, National Hospital Organization Minami-Okayama Medical Center kn-affil= affil-num=6 en-affil=Department of Gastroenterology, Okayama City Hospital kn-affil= affil-num=7 en-affil=Department of Internal Medicine, Fukuyama City Hospital kn-affil= affil-num=8 en-affil=Department of Respiratory Medicine, National Hospital Organization Minami-Okayama Medical Center kn-affil= affil-num=9 en-affil=Department of Allergy and Respiratory Medicine, National Hospital Organization Minami-Okayama Medical Center kn-affil= affil-num=10 en-affil=Department of Infectious Disease, Okayama City Hospital kn-affil= affil-num=11 en-affil=Department of Internal Medicine, Fukuyama City Hospital kn-affil= affil-num=12 en-affil=Center for Innovative Clinical Medicine, Okayama University Hospital kn-affil= affil-num=13 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= affil-num=14 en-affil=Department of Emergency, Critical Care and Disaster Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=15 en-affil=Department of Emergency, Critical Care and Disaster Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=16 en-affil=Department of General Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=17 en-affil=Department of General Thoracic Surgery and Breast and Endocrine Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=18 en-affil=Department of Pediatrics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=19 en-affil=Department of Hematology, Oncology and Respiratory Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=novel coronavirus disease 2019 kn-keyword=novel coronavirus disease 2019 en-keyword=pneumonia kn-keyword=pneumonia en-keyword=hydroperoxide kn-keyword=hydroperoxide en-keyword=nitric oxide kn-keyword=nitric oxide en-keyword=reactive oxygen species kn-keyword=reactive oxygen species en-keyword=citrulline kn-keyword=citrulline en-keyword=arginine kn-keyword=arginine en-keyword=asymmetric dimethylarginine kn-keyword=asymmetric dimethylarginine END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=“œ”A•a‚Ι‚¨‚―‚ι‹ίˆΚ”AΧŠΗΧ–EƒŠƒ\ƒ\[ƒ€‹@”\αŠQ‚ƁAƒAƒ~ƒmŽ_—¬“ό‘jŠQ‚Ι‚ζ‚ι‰ό‘PŒψ‰Κ kn-title=Inhibition of?amino acids influx into proximal tubular cells improves lysosome function in diabetes en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=KANOYuzuki en-aut-sei=KANO en-aut-mei=Yuzuki kn-aut-name=‰Α”[‹|ŒŽ kn-aut-sei=‰Α”[ kn-aut-mei=‹|ŒŽ aut-affil-num=1 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil=‰ͺŽR‘εŠw‘εŠw‰@ˆγŽ•–ςŠw‘‡Œ€‹†‰Θ END start-ver=1.4 cd-journal=joma no-vol=174 cd-vols= no-issue=5 article-no= start-page=451 end-page=459 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230801 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Direct binding of calmodulin to the cytosolic C-terminal regions of sweet/umami taste receptors en-subtitle= kn-subtitle= en-abstract= kn-abstract=Sweet and umami taste receptors recognize chemicals such as sugars and amino acids on their extracellular side and transmit signals into the cytosol of the taste cell. In contrast to ligands that act on the extracellular side of these receptors, little is known regarding the molecules that regulate receptor functions within the cytosol. In this study, we analysed the interaction between sweet and umami taste receptors and calmodulin, a representative Ca2+-dependent cytosolic regulatory protein. High prediction scores for calmodulin binding were observed on the C-terminal cytosolic side of mouse taste receptor type 1 subunit 3 (T1r3), a subunit that is common to both sweet and umami taste receptors. Pull-down assay and surface plasmon resonance analyses showed different affinities of calmodulin to the C-terminal tails of distinct T1r subtypes. Furthermore, we found that T1r3 and T1r2 showed the highest and considerable binding to calmodulin, whereas T1r1 showed weaker binding affinity. Finally, the binding of calmodulin to T1rs was consistently higher in the presence of Ca2+ than in its absence. The results suggested a possibility of the Ca2+-dependent feedback regulation process of sweet and umami taste receptor signaling by calmodulin. en-copyright= kn-copyright= en-aut-name=YoshidaAtsuki en-aut-sei=Yoshida en-aut-mei=Atsuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ItoAyumi en-aut-sei=Ito en-aut-mei=Ayumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=calmodulin kn-keyword=calmodulin en-keyword=cytosol kn-keyword=cytosol en-keyword=sweet taste kn-keyword=sweet taste en-keyword=taste receptor type 1 kn-keyword=taste receptor type 1 en-keyword=umami taste kn-keyword=umami taste END start-ver=1.4 cd-journal=joma no-vol=46 cd-vols= no-issue=4 article-no= start-page=180 end-page=186 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=202404 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Exploration of urine metabolic biomarkers for new-onset, untreated pediatric epilepsy: A gas and liquid chromatography mass spectrometry-based metabolomics study en-subtitle= kn-subtitle= en-abstract= kn-abstract=Objective: The discovery of objective indicators for recent epileptic seizures will help confirm the diagnosis of epilepsy and evaluate therapeutic effects. Past studies had shortcomings such as the inclusion of patients under treatment and those with various etiologies that could confound the analysis results significantly. We aimed to minimize such confounding effects and to explore the small molecule biomarkers associated with the recent occurrence of epileptic seizures using urine metabolomics.
Methods: This is a multicenter prospective study. Subjects included pediatric patients aged 2 to 12 years old with new-onset, untreated epilepsy, who had had the last seizure within 1 month before urine collection. Controls included healthy children aged 2 to 12 years old. Those with underlying or chronic diseases, acute illnesses, or recent administration of medications or supplements were excluded. Targeted metabolome analysis of spot urine samples was conducted using gas chromatography (GC)- and liquid chromatography (LC)-tandem mass spectrometry (MS/MS).
Results: We enrolled 17 patients and 21 controls. Among 172 metabolites measured by GC/MS/MS and 41 metabolites measured by LC/MS/MS, only taurine was consistently reduced in the epilepsy group. This finding was subsequently confirmed by the absolute quantification of amino acids. No other metabolites were consistently altered between the two groups.
Conclusions: Urine metabolome analysis, which covers a larger number of metabolites than conventional biochemistry analyses, found no consistently altered small molecule metabolites except for reduced taurine in epilepsy patients compared to healthy controls. Further studies with larger samples, subjects with different ages, expanded target metabolites, and the investigation of plasma samples are required. en-copyright= kn-copyright= en-aut-name=AkiyamaTomoyuki en-aut-sei=Akiyama en-aut-mei=Tomoyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SaigusaDaisuke en-aut-sei=Saigusa en-aut-mei=Daisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=InoueTakushi en-aut-sei=Inoue en-aut-mei=Takushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TokorodaniChiho en-aut-sei=Tokorodani en-aut-mei=Chiho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=AkiyamaMari en-aut-sei=Akiyama en-aut-mei=Mari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MichiueRie en-aut-sei=Michiue en-aut-mei=Rie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MoriAtsushi en-aut-sei=Mori en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=HishinumaEiji en-aut-sei=Hishinuma en-aut-mei=Eiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=MatsukawaNaomi en-aut-sei=Matsukawa en-aut-mei=Naomi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=ShibataTakashi en-aut-sei=Shibata en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TsuchiyaHiroki en-aut-sei=Tsuchiya en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=KobayashiKatsuhiro en-aut-sei=Kobayashi en-aut-mei=Katsuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= affil-num=1 en-affil=Department of Pediatrics (Child Neurology), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Laboratory of Biomedical and Analytical Sciences, Faculty of Pharma-Science, Teikyo University kn-affil= affil-num=3 en-affil=Department of Pediatric Neurology, NHO Okayama Medical Center kn-affil= affil-num=4 en-affil=Department of Pediatrics, Kochi Health Sciences Center kn-affil= affil-num=5 en-affil=Department of Pediatrics (Child Neurology), Okayama University Hospital kn-affil= affil-num=6 en-affil=Department of Pediatrics (Child Neurology), Okayama University Hospital kn-affil= affil-num=7 en-affil=Department of Neurology, Shiga Medical Center for Children kn-affil= affil-num=8 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=9 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=10 en-affil=Department of Pediatrics (Child Neurology), Okayama University Hospital kn-affil= affil-num=11 en-affil=Department of Pediatrics (Child Neurology), Okayama University Hospital kn-affil= affil-num=12 en-affil=Department of Pediatrics (Child Neurology), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=Amino acids kn-keyword=Amino acids en-keyword=Gas chromatography kn-keyword=Gas chromatography en-keyword=Liquid chromatography kn-keyword=Liquid chromatography en-keyword=Mass spectrometry kn-keyword=Mass spectrometry en-keyword=New-onset epilepsy kn-keyword=New-onset epilepsy END start-ver=1.4 cd-journal=joma no-vol=19 cd-vols= no-issue=4 article-no= start-page=e0300634 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240426 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Overexpression of the flagellar motor protein MotB sensitizes Bacillus subtilis to aminoglycosides in a motility-independent manner en-subtitle= kn-subtitle= en-abstract= kn-abstract=The flagellar motor proteins, MotA and MotB, form a complex that rotates the flagella by utilizing the proton motive force (PMF) at the bacterial cell membrane. Although PMF affects the susceptibility to aminoglycosides, the effect of flagellar motor proteins on the susceptibility to aminoglycosides has not been investigated. Here, we found that MotB overexpression increased susceptibility to aminoglycosides, such as kanamycin and gentamicin, in Bacillus subtilis without affecting swimming motility. MotB overexpression did not affect susceptibility to ribosome-targeting antibiotics other than aminoglycosides, cell wall-targeting antibiotics, DNA synthesis-inhibiting antibiotics, or antibiotics inhibiting RNA synthesis. Meanwhile, MotB overexpression increased the susceptibility to aminoglycosides even in the motA-deletion mutant, which lacks swimming motility. Overexpression of the MotB mutant protein carrying an amino acid substitution at the proton-binding site (D24A) resulted in the loss of the enhanced aminoglycoside-sensitive phenotype. These results suggested that MotB overexpression sensitizes B. subtilis to aminoglycosides in a motility-independent manner. Notably, the aminoglycoside-sensitive phenotype induced by MotB requires the proton-binding site but not the MotA/MotB complex formation. en-copyright= kn-copyright= en-aut-name=UnemeMio en-aut-sei=Uneme en-aut-mei=Mio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IshikawaKazuya en-aut-sei=Ishikawa en-aut-mei=Kazuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=FurutaKazuyuki en-aut-sei=Furuta en-aut-mei=Kazuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KaitoChikara en-aut-sei=Kaito en-aut-mei=Chikara kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=52 cd-vols= no-issue=10 article-no= start-page=5825 end-page=5840 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240425 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The ABCF proteins in Escherichia coli individually cope with 'hard-to-translate' nascent peptide sequences en-subtitle= kn-subtitle= en-abstract= kn-abstract=Organisms possess a wide variety of proteins with diverse amino acid sequences, and their synthesis relies on the ribosome. Empirical observations have led to the misconception that ribosomes are robust protein factories, but in reality, they have several weaknesses. For instance, ribosomes stall during the translation of the proline-rich sequences, but the elongation factor EF-P assists in synthesizing proteins containing the poly-proline sequences. Thus, living organisms have evolved to expand the translation capability of ribosomes through the acquisition of translation elongation factors. In this study, we have revealed that Escherichia coli ATP-Binding Cassette family-F (ABCF) proteins, YheS, YbiT, EttA and Uup, individually cope with various problematic nascent peptide sequences within the exit tunnel. The correspondence between noncanonical translations and ABCFs was YheS for the translational arrest by nascent SecM, YbiT for poly-basic sequence-dependent stalling and poly-acidic sequence-dependent intrinsic ribosome destabilization (IRD), EttA for IRD at the early stage of elongation, and Uup for poly-proline-dependent stalling. Our results suggest that ATP hydrolysis-coupled structural rearrangement and the interdomain linker sequence are pivotal for handling 'hard-to-translate' nascent peptides. Our study highlights a new aspect of ABCF proteins to reduce the potential risks that are encoded within the nascent peptide sequences. Graphical Abstract en-copyright= kn-copyright= en-aut-name=ChadaniYuhei en-aut-sei=Chadani en-aut-mei=Yuhei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamanouchiShun en-aut-sei=Yamanouchi en-aut-mei=Shun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=UemuraEri en-aut-sei=Uemura en-aut-mei=Eri kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamasakiKohei en-aut-sei=Yamasaki en-aut-mei=Kohei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=NiwaTatsuya en-aut-sei=Niwa en-aut-mei=Tatsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=IkedaToma en-aut-sei=Ikeda en-aut-mei=Toma kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KuriharaMiku en-aut-sei=Kurihara en-aut-mei=Miku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=IwasakiWataru en-aut-sei=Iwasaki en-aut-mei=Wataru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=TaguchiHideki en-aut-sei=Taguchi en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Faculty of Environmental, Life, Natural Science and Technology, Okayama University kn-affil= affil-num=2 en-affil=Department of Biological Sciences, Graduate School of Science, the University of Tokyo kn-affil= affil-num=3 en-affil=Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology kn-affil= affil-num=4 en-affil=Faculty of Science, Okayama University kn-affil= affil-num=5 en-affil=Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology kn-affil= affil-num=6 en-affil=School of Life Science and Technology, Tokyo Institute of Technology kn-affil= affil-num=7 en-affil=School of Life Science and Technology, Tokyo Institute of Technology kn-affil= affil-num=8 en-affil=Department of Biological Sciences, Graduate School of Science, the University of Tokyo kn-affil= affil-num=9 en-affil=Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology kn-affil= END start-ver=1.4 cd-journal=joma no-vol=19 cd-vols= no-issue=3 article-no= start-page=e0300981 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240322 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Chemical range recognized by the ligand-binding domain in a representative amino acid-sensing taste receptor, T1r2a/T1r3, from medaka fish en-subtitle= kn-subtitle= en-abstract= kn-abstract=Taste receptor type 1 (T1r) proteins are responsible for recognizing nutrient chemicals in foods. In humans, T1r2/T1r3 and T1r1/T1r3 heterodimers serve as the sweet and umami receptors that recognize sugars or amino acids and nucleotides, respectively. T1rs are conserved among vertebrates, and T1r2a/T1r3 from medaka fish is currently the only member for which the structure of the ligand-binding domain (LBD) has been solved. T1r2a/T1r3 is an amino acid receptor that recognizes various l-amino acids in its LBD as observed with other T1rs exhibiting broad substrate specificities. Nevertheless, the range of chemicals that are recognized by T1r2a/T1r3LBD has not been extensively explored. In the present study, the binding of various chemicals to medaka T1r2a/T1r3LBD was analyzed. A binding assay for amino acid derivatives verified the specificity of this protein to l-alpha-amino acids and the importance of alpha-amino and carboxy groups for receptor recognition. The results further indicated the significance of the alpha-hydrogen for recognition as replacing it with a methyl group resulted in a substantially decreased affinity. The binding ability to the protein was not limited to proteinogenic amino acids, but also to non-proteinogenic amino acids, such as metabolic intermediates. Besides l-alpha-amino acids, no other chemicals showed significant binding to the protein. These results indicate that all of the common structural groups of alpha-amino acids and their geometry in the l-configuration are recognized by the protein, whereas a wide variety of alpha-substituents can be accommodated in the ligand binding sites of the LBDs. en-copyright= kn-copyright= en-aut-name=IshidaHikaru en-aut-sei=Ishida en-aut-mei=Hikaru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=78 cd-vols= no-issue=2 article-no= start-page=95 end-page=106 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=202404 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The Roles of Neuropeptide Y in Respiratory Disease Pathogenesis via the Airway Immune Response en-subtitle= kn-subtitle= en-abstract= kn-abstract=The lungs are very complex organs, and the respiratory system performs the dual roles of repairing tissue while protecting against infection from various environmental stimuli. Persistent external irritation disrupts the immune responses of tissues and cells in the respiratory system, ultimately leading to respiratory disease. Neuropeptide Y (NPY) is a 36-amino-acid polypeptide and a neurotransmitter that regulates homeostasis. The NPY receptor is a seven-transmembrane-domain G-protein-coupled receptor with six subtypes (Y1, Y2, Y3, Y4, Y5, and Y6). Of these receptors, Y1, Y2, Y4, and Y5 are functional in humans, and Y1 plays important roles in the immune responses of many organs, including the respiratory system. NPY and the Y1 receptor have critical roles in the pathogenesis of asthma, chronic obstructive pulmonary disease, and idiopathic pulmonary fibrosis. The effects of NPY on the airway immune response and pathogenesis differ among respiratory diseases. This review focuses on the involvement of NPY in the airway immune response and pathogenesis of various respiratory diseases. en-copyright= kn-copyright= en-aut-name=ItanoJunko en-aut-sei=Itano en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KiuraKatsuyuki en-aut-sei=Kiura en-aut-mei=Katsuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MaedaYoshinobu en-aut-sei=Maeda en-aut-mei=Yoshinobu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MiyaharaNobuaki en-aut-sei=Miyahara en-aut-mei=Nobuaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Hematology, Oncology and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= affil-num=3 en-affil=Department of Hematology, Oncology and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= en-keyword=neuropeptide y kn-keyword=neuropeptide y en-keyword=Y1 receptor kn-keyword=Y1 receptor en-keyword=airway immune response kn-keyword=airway immune response en-keyword=bronchial epithelial cells kn-keyword=bronchial epithelial cells en-keyword=respiratory disease kn-keyword=respiratory disease END start-ver=1.4 cd-journal=joma no-vol=5 cd-vols= no-issue=2 article-no= start-page=182 end-page=194 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20231208 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Inhibition of Amino Acids Influx into Proximal Tubular Cells Improves Lysosome Function in Diabetes en-subtitle= kn-subtitle= en-abstract= kn-abstract=Background Inhibition of glucose influx into proximal tubular cells (PTCs) by sodium?glucose cotransporter 2 inhibitors revealed prominent therapeutic effects on diabetic kidney disease. Collectrin (CLTRN) serves as a chaperone for the trafficking of neutral amino acid (AA) transporters in the apical membranes of PTCs. We investigated the beneficial effects of reduced influx of AAs into PTCs in diabetes and obesity model of Cltrn?/y mice.
Methods Cltrn+/y and Cltrn?/y mice at age 5 weeks were assigned to standard diet and streptozotocin and high-fat diet (STZ-HFD)?treated groups.
Results At age 22?23 weeks, body weight and HbA1c levels significantly increased in STZ-HFD-Cltrn+/y compared with standard diet-Cltrn+/y; however, they were not altered in STZ-HFD-Cltrn?/y compared with STZ-HFD-Cltrn+/y. At age 20 weeks, urinary albumin creatinine ratio was significantly reduced in STZ-HFD-Cltrn?/y compared with STZ-HFD-Cltrn+/y. Under the treatments with STZ and HFD, the Cltrn gene deficiency caused significant increase in urinary concentration of AAs such as Gln, His, Gly, Thr, Tyr, Val, Trp, Phe, Ile, Leu, and Pro. In PTCs in STZ-HFD-Cltrn+/y, the enlarged lysosomes with diameter of 10 ƒΚm or more were associated with reduced autolysosomes, and the formation of giant lysosomes was prominently suppressed in STZ-HFD-Cltrn?/y. Phospho-mTOR and inactive form of phospho-transcription factor EB were reduced in STZ-HFD-Cltrn?/y compared with STZ-HFD-Cltrn+/y.
Conclusions The reduction of AAs influx into PTCs inactivated mTOR, activated transcription factor EB, improved lysosome function, and ameliorated vacuolar formation of PTCs in STZ-HFD-Cltrn?/y mice. en-copyright= kn-copyright= en-aut-name=KanoYuzuki en-aut-sei=Kano en-aut-mei=Yuzuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamaguchiSatoshi en-aut-sei=Yamaguchi en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MiseKoki en-aut-sei=Mise en-aut-mei=Koki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KawakitaChieko en-aut-sei=Kawakita en-aut-mei=Chieko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OnishiYasuhiro en-aut-sei=Onishi en-aut-mei=Yasuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KurookaNaoko en-aut-sei=Kurooka en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SugawaraRyosuke en-aut-sei=Sugawara en-aut-mei=Ryosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=AlbuayjanHaya Hamed Hassan en-aut-sei=Albuayjan en-aut-mei=Haya Hamed Hassan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=NakatsukaAtsuko en-aut-sei=Nakatsuka en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=EguchiJun en-aut-sei=Eguchi en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=WadaJun en-aut-sei=Wada en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=6 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=7 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=8 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=9 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=10 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=11 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=diabetes mellitus kn-keyword=diabetes mellitus en-keyword=diabetic nephropathy kn-keyword=diabetic nephropathy en-keyword=metabolism kn-keyword=metabolism en-keyword=obesity kn-keyword=obesity en-keyword=tubular epithelium kn-keyword=tubular epithelium END start-ver=1.4 cd-journal=joma no-vol=626 cd-vols= no-issue=7999 article-no= start-page=670 end-page=677 dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240131 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Oxygen-evolving photosystem II structures during S1?S2?S3 transitions en-subtitle= kn-subtitle= en-abstract= kn-abstract=Photosystem II (PSII) catalyses the oxidation of water through a four-step cycle of Si states (i?=?0?4) at the Mn4CaO5 cluster1,2,3, during which an extra oxygen (O6) is incorporated at the S3 state to form a possible dioxygen4,5,6,7. Structural changes of the metal cluster and its environment during the S-state transitions have been studied on the microsecond timescale. Here we use pump-probe serial femtosecond crystallography to reveal the structural dynamics of PSII from nanoseconds to milliseconds after illumination with one flash (1F) or two flashes (2F). YZ, a tyrosine residue that connects the reaction centre P680 and the Mn4CaO5 cluster, showed structural changes on a nanosecond timescale, as did its surrounding amino acid residues and water molecules, reflecting the fast transfer of electrons and protons after flash illumination. Notably, one water molecule emerged in the vicinity of Glu189 of the D1 subunit of PSII (D1-E189), and was bound to the Ca2+ ion on a sub-microsecond timescale after 2F illumination. This water molecule disappeared later with the concomitant increase of O6, suggesting that it is the origin of O6. We also observed concerted movements of water molecules in the O1, O4 and Cl-1 channels and their surrounding amino acid residues to complete the sequence of electron transfer, proton release and substrate water delivery. These results provide crucial insights into the structural dynamics of PSII during S-state transitions as well as O?O bond formation. en-copyright= kn-copyright= en-aut-name=LiHongjie en-aut-sei=Li en-aut-mei=Hongjie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NakajimaYoshiki en-aut-sei=Nakajima en-aut-mei=Yoshiki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NangoEriko en-aut-sei=Nango en-aut-mei=Eriko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OwadaShigeki en-aut-sei=Owada en-aut-mei=Shigeki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamadaDaichi en-aut-sei=Yamada en-aut-mei=Daichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HashimotoKana en-aut-sei=Hashimoto en-aut-mei=Kana kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=LuoFangjia en-aut-sei=Luo en-aut-mei=Fangjia kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TanakaRie en-aut-sei=Tanaka en-aut-mei=Rie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=AkitaFusamichi en-aut-sei=Akita en-aut-mei=Fusamichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=KatoKoji en-aut-sei=Kato en-aut-mei=Koji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=KangJungmin en-aut-sei=Kang en-aut-mei=Jungmin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=SaitohYasunori en-aut-sei=Saitoh en-aut-mei=Yasunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=KishiShunpei en-aut-sei=Kishi en-aut-mei=Shunpei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=YuHuaxin en-aut-sei=Yu en-aut-mei=Huaxin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=MatsubaraNaoki en-aut-sei=Matsubara en-aut-mei=Naoki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= en-aut-name=FujiiHajime en-aut-sei=Fujii en-aut-mei=Hajime kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=16 ORCID= en-aut-name=SugaharaMichihiro en-aut-sei=Sugahara en-aut-mei=Michihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=17 ORCID= en-aut-name=SuzukiMamoru en-aut-sei=Suzuki en-aut-mei=Mamoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=18 ORCID= en-aut-name=MasudaTetsuya en-aut-sei=Masuda en-aut-mei=Tetsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=19 ORCID= en-aut-name=KimuraTetsunari en-aut-sei=Kimura en-aut-mei=Tetsunari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=20 ORCID= en-aut-name=ThaoTran Nguyen en-aut-sei=Thao en-aut-mei=Tran Nguyen kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=21 ORCID= en-aut-name=YonekuraShinichiro en-aut-sei=Yonekura en-aut-mei=Shinichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=22 ORCID= en-aut-name=YuLong-Jiang en-aut-sei=Yu en-aut-mei=Long-Jiang kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=23 ORCID= en-aut-name=ToshaTakehiko en-aut-sei=Tosha en-aut-mei=Takehiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=24 ORCID= en-aut-name=TonoKensuke en-aut-sei=Tono en-aut-mei=Kensuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=25 ORCID= en-aut-name=JotiYasumasa en-aut-sei=Joti en-aut-mei=Yasumasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=26 ORCID= en-aut-name=HatsuiTakaki en-aut-sei=Hatsui en-aut-mei=Takaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=27 ORCID= en-aut-name=YabashiMakina en-aut-sei=Yabashi en-aut-mei=Makina kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=28 ORCID= en-aut-name=KuboMinoru en-aut-sei=Kubo en-aut-mei=Minoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=29 ORCID= en-aut-name=IwataSo en-aut-sei=Iwata en-aut-mei=So kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=30 ORCID= en-aut-name=IsobeHiroshi en-aut-sei=Isobe en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=31 ORCID= en-aut-name=YamaguchiKizashi en-aut-sei=Yamaguchi en-aut-mei=Kizashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=32 ORCID= en-aut-name=SugaMichihiro en-aut-sei=Suga en-aut-mei=Michihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=33 ORCID= en-aut-name=ShenJian-Ren en-aut-sei=Shen en-aut-mei=Jian-Ren kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=34 ORCID= affil-num=1 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=2 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=3 en-affil=Institute of Multidisciplinary Research for Advanced Materials, Tohoku University kn-affil= affil-num=4 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=5 en-affil=Department of Picobiology, Graduate School of Life Science, University of Hyogo kn-affil= affil-num=6 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=7 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=8 en-affil=RIKEN SPring-8 Center kn-affil= affil-num=9 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=10 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=11 en-affil=RIKEN SPring-8 Center kn-affil= affil-num=12 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=13 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=14 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=15 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=16 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=17 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=18 en-affil=Institute for Protein Research, Osaka University kn-affil= affil-num=19 en-affil=Division of Food and Nutrition, Faculty of Agriculture, Ryukoku University kn-affil= affil-num=20 en-affil=Department of Chemistry, Graduate School of Science, Kobe University kn-affil= affil-num=21 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=22 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=23 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=24 en-affil=RIKEN SPring-8 Center kn-affil= affil-num=25 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=26 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=27 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=28 en-affil=Japan Synchrotron Radiation Research Institute kn-affil= affil-num=29 en-affil=Department of Picobiology, Graduate School of Life Science, University of Hyogo kn-affil= affil-num=30 en-affil=RIKEN SPring-8 Center kn-affil= affil-num=31 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=32 en-affil=Center for Quantum Information and Quantum Biology, Osaka University kn-affil= affil-num=33 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=34 en-affil=Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=81 cd-vols= no-issue=3 article-no= start-page=80 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240128 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Mutational analysis of the transmembrane ƒΏ4-helix of Bacillus thuringiensis mosquito-larvicidal Cry4Aa toxin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Cry4Aa, produced by Bacillus thuringiensis subsp. israelensis, exhibits specific toxicity to larvae of medically important mosquito genera. Cry4Aa functions as a pore-forming toxin, and a helical hairpin (ƒΏ4-loop-ƒΏ5) of domain I is believed to be the transmembrane domain that forms toxin pores. Pore formation is considered to be a central mode of Cry4Aa action, but the relationship between pore formation and toxicity is poorly understood. In the present study, we constructed Cry4Aa mutants in which each polar amino acid residues within the transmembrane ƒΏ4 helix was replaced with glutamic acid. Bioassays using Culex pipiens mosquito larvae and subsequent ion permeability measurements using symmetric KCl solution revealed an apparent correlation between toxicity and toxin pore conductance for most of the Cry4Aa mutants. In contrast, the Cry4Aa mutant H178E was a clear exception, almost losing its toxicity but still exhibiting a moderately high conductivity of about 60% of the wild-type. Furthermore, the conductance of the pore formed by the N190E mutant (about 50% of the wild-type) was close to that of H178E, but the toxicity was significantly higher than that of H178E. Ion selectivity measurements using asymmetric KCl solution revealed a significant decrease in cation selectivity of toxin pores formed by H178E compared to N190E. Our data suggest that the toxicity of Cry4Aa is primarily pore related. The formation of toxin pores that are highly ion-permeable and also highly cation-selective may enhance the influx of cations and water into the target cell, thereby facilitating the eventual death of mosquito larvae. en-copyright= kn-copyright= en-aut-name=TakahashiHirokazu en-aut-sei=Takahashi en-aut-mei=Hirokazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AsakuraMami en-aut-sei=Asakura en-aut-mei=Mami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IdeToru en-aut-sei=Ide en-aut-mei=Toru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HayakawaTohru en-aut-sei=Hayakawa en-aut-mei=Tohru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=220 cd-vols= no-issue=2 article-no= start-page=16 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2024 dt-pub=20240108 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Tamyb10-D1 restores red grain color and increases grain dormancy via suppressing expression of TaLTP2.128, non-specific lipid transfer protein in wheat en-subtitle= kn-subtitle= en-abstract= kn-abstract=Grain dormancy of wheat is closely associated with grain color: red-grained lines show higher dormancy than white-grained lines. The production of red pigments is regulated by R-1, Tamyb10 gene. However, the relation between grain color and dormancy remains unknown. For this study, we generated transgenic lines which were introduced a DNA fragment containing Tamyb10-D1 gene and its a 2 kb promoter including the 5Œ untranslated region into white-grained wheat. Transgenic lines showed red-grained and higher dormant traits. Contents of plant hormones and gene expression of embryos at 30 days after pollination were examined in a wild type and a transgenic line. No differences were observed in the contents of plant hormones, but several genes are differentially expressed between these lines. One differentially expressed gene, TaLTP2.128, is a member of non-specific lipid transfer proteins. It was expressed higher in white grains than in red grains. A putative amino acid sequence showed similarity to that of OsHyPRP5, which is identified as QTL controlling low-temperature germinability in rice. Expression of TaLTP2.128 was increased by grain imbibition. The increasing levels were higher not only in other white-grained lines, but also in non-dormant red-grained lines. TaLTP2.128 was expressed at a quite early stage of germination. These study findings indicate that Tamyb10 regulates dormancy release by the modification of TaLTP2.128 acting as trigger of germination. en-copyright= kn-copyright= en-aut-name=HimiEiko en-aut-sei=Himi en-aut-mei=Eiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=Kurihara-YonemotoShiho en-aut-sei=Kurihara-Yonemoto en-aut-mei=Shiho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AbeFumitaka en-aut-sei=Abe en-aut-mei=Fumitaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakahashiHidekazu en-aut-sei=Takahashi en-aut-mei=Hidekazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TanakaKeisuke en-aut-sei=Tanaka en-aut-mei=Keisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MatsuuraTakakazu en-aut-sei=Matsuura en-aut-mei=Takakazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MaekawaMasahiko en-aut-sei=Maekawa en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=SasakiTakuji en-aut-sei=Sasaki en-aut-mei=Takuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=RikiishiKazuhide en-aut-sei=Rikiishi en-aut-mei=Kazuhide kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Kibi International University kn-affil= affil-num=2 en-affil=Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization kn-affil= affil-num=3 en-affil=Institute of Crop Science, National Agriculture and Food Research Organization kn-affil= affil-num=4 en-affil=Fukushima University kn-affil= affil-num=5 en-affil=NODAI Genome Research Center, Tokyo University of Agriculture kn-affil= affil-num=6 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=7 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=8 en-affil=NODAI Research Institute, Tokyo University of Agriculture kn-affil= affil-num=9 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=Lipid transfer protein kn-keyword=Lipid transfer protein en-keyword=Pre-harvest sprouting kn-keyword=Pre-harvest sprouting en-keyword=Seed dormancy kn-keyword=Seed dormancy en-keyword=Seed germination kn-keyword=Seed germination en-keyword=Tamyb10 kn-keyword=Tamyb10 en-keyword=Wheat kn-keyword=Wheat END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue= article-no= start-page=RP88822 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20231121 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Characterization of tryptophan oxidation affecting D1 degradation by FtsH in the photosystem II quality control of chloroplasts en-subtitle= kn-subtitle= en-abstract= kn-abstract=Photosynthesis is one of the most important reactions for sustaining our environment. Photosystem II (PSII) is the initial site of photosynthetic electron transfer by water oxidation. Light in excess, however, causes the simultaneous production of reactive oxygen species (ROS), leading to photo-oxidative damage in PSII. To maintain photosynthetic activity, the PSII reaction center protein D1, which is the primary target of unavoidable photo-oxidative damage, is efficiently degraded by FtsH protease. In PSII subunits, photo-oxidative modifications of several amino acids such as Trp have been indeed documented, whereas the linkage between such modifications and D1 degradation remains elusive. Here, we show that an oxidative post-translational modification of Trp residue at the N-terminal tail of D1 is correlated with D1 degradation by FtsH during high-light stress. We revealed that Arabidopsis mutant lacking FtsH2 had increased levels of oxidative Trp residues in D1, among which an N-terminal Trp-14 was distinctively localized in the stromal side. Further characterization of Trp-14 using chloroplast transformation in Chlamydomonas indicated that substitution of D1 Trp-14 to Phe, mimicking Trp oxidation enhanced FtsH-mediated D1 degradation under high light, although the substitution did not affect protein stability and PSII activity. Molecular dynamics simulation of PSII implies that both Trp-14 oxidation and Phe substitution cause fluctuation of D1 N-terminal tail. Furthermore, Trp-14 to Phe modification appeared to have an additive effect in the interaction between FtsH and PSII core in vivo. Together, our results suggest that the Trp oxidation at its N-terminus of D1 may be one of the key oxidations in the PSII repair, leading to processive degradation by FtsH. en-copyright= kn-copyright= en-aut-name=KatoYusuke en-aut-sei=Kato en-aut-mei=Yusuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KurodaHiroshi en-aut-sei=Kuroda en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OzawaShin-Ichiro en-aut-sei=Ozawa en-aut-mei=Shin-Ichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SaitoKeisuke en-aut-sei=Saito en-aut-mei=Keisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=DograVivek en-aut-sei=Dogra en-aut-mei=Vivek kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=ScholzMartin en-aut-sei=Scholz en-aut-mei=Martin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=ZhangGuoxian en-aut-sei=Zhang en-aut-mei=Guoxian kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=de VitryCatherine en-aut-sei=de Vitry en-aut-mei=Catherine kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=IshikitaHiroshi en-aut-sei=Ishikita en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=KimChanhong en-aut-sei=Kim en-aut-mei=Chanhong kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=HipplerMichael en-aut-sei=Hippler en-aut-mei=Michael kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=TakahashiYuichiro en-aut-sei=Takahashi en-aut-mei=Yuichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=SakamotoWataru en-aut-sei=Sakamoto en-aut-mei=Wataru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=2 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=4 en-affil=Research Center for Advanced Science and Technology, The University of Tokyo kn-affil= affil-num=5 en-affil=Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences kn-affil= affil-num=6 en-affil=Institute of Plant Biology and Biotechnology, University of M?nster kn-affil= affil-num=7 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=8 en-affil=Institut de Biologie Physico-Chimique, Unit? Mixte de Recherche 7141, Centre National de la Recherche Scientifique and Sorbonne Universit? Pierre et Marie Curie kn-affil= affil-num=9 en-affil=Research Center for Advanced Science and Technology, The University of Tokyo kn-affil= affil-num=10 en-affil=Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences kn-affil= affil-num=11 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=12 en-affil=Research Institute for Interdisciplinary Science, Okayama University kn-affil= affil-num=13 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= en-keyword=post-translational modification kn-keyword=post-translational modification en-keyword=Arabidopsis thaliana kn-keyword=Arabidopsis thaliana en-keyword=protein degradation kn-keyword=protein degradation en-keyword=photosystem II kn-keyword=photosystem II en-keyword=photo-oxidative damage kn-keyword=photo-oxidative damage en-keyword=tryptophan oxidation kn-keyword=tryptophan oxidation en-keyword=Chlamydomonas reinhardtii kn-keyword=Chlamydomonas reinhardtii END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue=12 article-no= start-page=1481 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20231201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Random Mutational Analysis Targeting Residue K155 within the Transmembrane ƒΐ-Hairpin of the Mosquitocidal Mpp46Ab Toxin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Mpp46Ab is a mosquito-larvicidal pore-forming toxin derived from Bacillus thuringiensis TK-E6. Pore formation is believed to be a central mode of Mpp46Ab action, and the cation selectivity of the channel pores, in particular, is closely related to its mosquito-larvicidal activity. In the present study, we constructed a mutant library in which residue K155 within the transmembrane ƒΐ-hairpin was randomly replaced with other amino acid residues. Upon mutagenesis and following primary screening using Culex pipiens mosquito larvae, we obtained 15 mutants in addition to the wild-type toxin. Bioassays using purified proteins revealed that two mutants, K155E and K155I, exhibited toxicity significantly higher than that of the wild-type toxin. Although increased cation selectivity was previously reported for K155E channel pores, we demonstrated in the present study that the cation selectivity of K155I channel pores was also significantly increased. Considering the characteristics of the amino acids, the charge of residue 155 may not directly affect the cation selectivity of Mpp46Ab channel pores. Replacement of K155 with glutamic acid or isoleucine may induce a similar conformational change in the region associated with the ion selectivity of the Mpp46Ab channel pores. Mutagenesis targeting the transmembrane ƒΐ-hairpin may be an effective strategy for enhancing the ion permeability of the channel pores and the resulting mosquito-larvicidal activity of Mpp46Ab. en-copyright= kn-copyright= en-aut-name=MiyazakiMidoka en-aut-sei=Miyazaki en-aut-mei=Midoka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AsakuraMami en-aut-sei=Asakura en-aut-mei=Mami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IdeToru en-aut-sei=Ide en-aut-mei=Toru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HayakawaTohru en-aut-sei=Hayakawa en-aut-mei=Tohru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= en-keyword=mosquito-larvicidal toxin kn-keyword=mosquito-larvicidal toxin en-keyword=Bacillus thuringiensis TK-E6 kn-keyword=Bacillus thuringiensis TK-E6 en-keyword=Culex pipiens mosquito larvae kn-keyword=Culex pipiens mosquito larvae en-keyword=site-directed mutagenesis kn-keyword=site-directed mutagenesis en-keyword=electrophysiologic analysis kn-keyword=electrophysiologic analysis END start-ver=1.4 cd-journal=joma no-vol=96 cd-vols= no-issue= article-no= start-page=129536 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20231115 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Direct evaluation of polarity of the ligand binding pocket in retinoid X receptor using a fluorescent solvatochromic agonist en-subtitle= kn-subtitle= en-abstract= kn-abstract=High selectivity of small-molecule drug candidates for their target molecule is important to minimize potential side effects. One factor that contributes to the selectivity is the internal polarity of the ligand-binding pocket (LBP) in the target molecule, but this is difficult to measure. Here, we first confirmed that the retinoid X receptor (RXR) agonist 6-(ethyl(1-isobutyl-2-oxo-4-(trifluoromethyl)-1,2-dihydroquinolin-7-yl)amino)nicotinic acid (NEt-iFQ, 1) exhibits fluorescence solvatochromism, i.e., its Stokes shift depends on the polarity of the solvent, and then we utilized this property to directly measure the internal polarity of the RXRƒΏ-LBP. The Stokes shift of 1 when bound to the RXRƒΏ-LBP corresponded to that of 1 in chloroform solution. This finding is expected to be helpful for designing RXR-selective ligands. A similar approach should be appliable to evaluate the internal polarity of the LBPs of other receptors. en-copyright= kn-copyright= en-aut-name=MiuraKizuku en-aut-sei=Miura en-aut-mei=Kizuku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=FujiharaMichiko en-aut-sei=Fujihara en-aut-mei=Michiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WatanabeMasaki en-aut-sei=Watanabe en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakamuraYuta en-aut-sei=Takamura en-aut-mei=Yuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KawasakiMayu en-aut-sei=Kawasaki en-aut-mei=Mayu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NakanoShogo en-aut-sei=Nakano en-aut-mei=Shogo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KakutaHiroki en-aut-sei=Kakuta en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Faculty of Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Division of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Division of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Division of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of Shizuoka kn-affil= affil-num=6 en-affil=Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of Shizuoka kn-affil= affil-num=7 en-affil=Division of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=RXR kn-keyword=RXR en-keyword=Fluorescence kn-keyword=Fluorescence en-keyword=Solvatochromism kn-keyword=Solvatochromism en-keyword=Binding assay kn-keyword=Binding assay END start-ver=1.4 cd-journal=joma no-vol=77 cd-vols= no-issue=6 article-no= start-page=635 end-page=645 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=202312 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effects of Nutritional Support Combined with Neuromuscular Electrical Stimulation on Muscle Strength and Thickness: A Randomized Controlled Trial in Healthy Young Adult Males en-subtitle= kn-subtitle= en-abstract= kn-abstract=In the management of post-injury patients with activity limitations, methods to prevent musculoskeletal disorders and hasten recovery are important. This randomized controlled, single-blinded study was a preliminary investigation of the combined effect of nutritional support with neuromuscular electrical stimulation (NMES) on muscle strength and thickness. Healthy young adult males (median age, 21 years) were enrolled; each of their hands was randomly assigned to one of the following four groups: Placebo, Nutrition, NMES, and Nutrition + NMES. All participants received whey protein or placebo (3x/week for 6 weeks) and NMES training (3x/week for 6 weeks) on the abductor digiti minimi (ADM) muscle of either the left or right hand. ADM muscle strength and thickness were analyzed at baseline and at week 7. We analyzed 38 hands (9 Placebo, 10 Nutrition, 9 NMES, 10 Nutrition + NMES). There was significantly greater muscle strengthening in the Nutrition + NMES group compared to the Placebo group or the NMES group, but no significant difference in gain of muscle thickness. The combined intervention may be effective in improving muscle strength. Future clinical trials targeting various muscles after sports-related injuries are warranted. en-copyright= kn-copyright= en-aut-name=IkedaTomohiro en-aut-sei=Ikeda en-aut-mei=Tomohiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OkamuraKazunori en-aut-sei=Okamura en-aut-mei=Kazunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HasegawaMasaki en-aut-sei=Hasegawa en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TanakaSatoshi en-aut-sei=Tanaka en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KanaiShusaku en-aut-sei=Kanai en-aut-mei=Shusaku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Department of Rehabilitation Medicine, Okayama University Hospital kn-affil= affil-num=2 en-affil=Department of Physical Therapy, Faculty of Health and Welfare, Prefectural University of Hiroshima kn-affil= affil-num=3 en-affil=Department of Physical Therapy, Faculty of Health and Welfare, Prefectural University of Hiroshima kn-affil= affil-num=4 en-affil=Department of Physical Therapy, Faculty of Health and Welfare, Prefectural University of Hiroshima kn-affil= affil-num=5 en-affil=Department of Physical Therapy, Faculty of Health and Welfare, Prefectural University of Hiroshima kn-affil= en-keyword=whey protein kn-keyword=whey protein en-keyword=electrical stimulation kn-keyword=electrical stimulation en-keyword=muscle strength kn-keyword=muscle strength en-keyword=healthy volunteers kn-keyword=healthy volunteers END start-ver=1.4 cd-journal=joma no-vol=32 cd-vols= no-issue=4 article-no= start-page=221 end-page=227 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=2023 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Novel and recurrent COMP gene variants in five Japanese patients with pseudoachondroplasia: skeletal changes from the neonatal to infantile periods en-subtitle= kn-subtitle= en-abstract= kn-abstract=Pseudoachondroplasia (PSACH) is an autosomal dominant skeletal dysplasia caused by pathogenic variants of cartilage oligomeric matrix protein (COMP). Clinical symptoms of PSACH are characterized by growth disturbances after the first year of life. These disturbances lead to severe short stature with short limbs, brachydactyly, scoliosis, joint laxity, joint pain since childhood, and a normal face. Epimetaphyseal dysplasia, shortened long bones, and short metacarpals and phalanges are common findings on radiological examination. Additionally, anterior tonguing of the vertebral bodies in the lateral view is an important finding in childhood because it is specific to PSACH and normalizes with age. Here, we report five Japanese patients with PSACH, with one recurrent (p.Cys351Tyr) and four novel heterozygous pathogenic COMP variants (p.Asp437Tyr, p.Asp446Gly, p.Asp507Tyr, and p.Asp518Val). These five pathogenic variants were located in the calcium-binding type 3 (T3) repeats. In four of the novel variants, the affected amino acid was aspartic acid, which is abundant in each of the eight T3 repeats. We describe the radiological findings of these five patients. We also retrospectively analyzed the sequential changes in the vertebral body and epimetaphysis of the long bones from the neonatal to infantile periods in a patient with PSACH and congenital heart disease. en-copyright= kn-copyright= en-aut-name=HasegawaKosei en-aut-sei=Hasegawa en-aut-mei=Kosei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=FutagawaNatsuko en-aut-sei=Futagawa en-aut-mei=Natsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AgoYuko en-aut-sei=Ago en-aut-mei=Yuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MiyaharaHiroyuki en-aut-sei=Miyahara en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=HaradaDaisuke en-aut-sei=Harada en-aut-mei=Daisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MiyazawaMari en-aut-sei=Miyazawa en-aut-mei=Mari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YoshimotoJunko en-aut-sei=Yoshimoto en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=BabaKenji en-aut-sei=Baba en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=MoriwakeTadashi en-aut-sei=Moriwake en-aut-mei=Tadashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=TanakaHiroyuki en-aut-sei=Tanaka en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TsukaharaHirokazu en-aut-sei=Tsukahara en-aut-mei=Hirokazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Department of Pediatrics, Okayama University Hospital kn-affil= affil-num=2 en-affil=Department of Pediatrics, Okayama University Hospital kn-affil= affil-num=3 en-affil=Department of Pediatrics, Okayama University Hospital kn-affil= affil-num=4 en-affil=Department of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Pediatrics, JCHO Osaka Hospital kn-affil= affil-num=6 en-affil=Department of Pediatrics, Kochi Health Sciences Center kn-affil= affil-num=7 en-affil=Department of Pediatrics, Okayama University Hospital kn-affil= affil-num=8 en-affil=Department of Pediatrics, Okayama University Hospital kn-affil= affil-num=9 en-affil=Department of Pediatrics, Iwakuni Clinical Center, National Hospital Organization kn-affil= affil-num=10 en-affil=Department of Pediatrics, Okayama Saiseikai General Hospital kn-affil= affil-num=11 en-affil=Department of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=infant kn-keyword=infant en-keyword=skeleton kn-keyword=skeleton en-keyword=spine kn-keyword=spine en-keyword=cartilage kn-keyword=cartilage en-keyword=growth kn-keyword=growth END start-ver=1.4 cd-journal=joma no-vol=42 cd-vols= no-issue=6 article-no= start-page=698 end-page=708 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230922 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Investigating the Effect of Substituting a Single Cysteine Residue on the Thermal Stability of an Engineered Sweet Protein, Single-Chain Monellin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Single-chain monellin (SCM) is an engineered protein that links the two chains of monellin, a naturally sweet-tasting protein. This protein is an attractive candidate for use as a sugar replacement in food and beverages and has numerous other applications. Therefore, generating SCM mutants with improved stability is an active area of research to broaden the range of its potential applications. In this study, we focused on the Cys41 residue of SCM, which is a single cysteine residue present at a structurally important position. This residue is often substituted with Ser. However, this substitution may destabilize SCM because Cys41 is buried in the hydrophobic core of the protein. Therefore, we designed mutants that substituted Ala, Val, and Leu for this residue, namely C41A, C41V, and C41L. We characterized these three mutants, SCM C41S, and wild type (WT). Differential scanning fluorimetric analysis revealed that substituting Cys41 with Ala or Val increased the thermal stability of SCM, while substitution with Ser or Leu decreased its stability. Determination of the crystal structures of SCM C41A and C41V mutants revealed that the overall structures and main chain structures around the 41st residue of both mutants were almost identical to the WT. On the other hand, the orientations of the amino acid side chains near the 41st residue differed among the SCM variants. Taken together, our results indicate that substituting Cys41 with Ala or Val increases the stability of SCM and provide insight into the structural basis of this improvement. en-copyright= kn-copyright= en-aut-name=OhnumaKyosuke en-aut-sei=Ohnuma en-aut-mei=Kyosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=School of Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=School of Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=School of Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=Crystallography kn-keyword=Crystallography en-keyword=Monellin kn-keyword=Monellin en-keyword=Protein Stability kn-keyword=Protein Stability en-keyword=Recombinant Proteins kn-keyword=Recombinant Proteins END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2005 dt-pub=20050930 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Œψ—¦‚Μ—Η‚’”ρ“V‘RƒAƒ~ƒmŽ_‚Μ“±“ό‚Μ‚½‚߂̐V‹KtRNA‚ΜŠJ”­ kn-title=Development of novel tRNAs for efficient incorporation of nonnatural amino acids en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=TairaHikaru en-aut-sei=Taira en-aut-mei=Hikaru kn-aut-name=•½—ΗŒυ kn-aut-sei=•½—Η kn-aut-mei=Œυ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2005 dt-pub=20050930 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=‘ε’°‹Ϋ—R—ˆ‚ΜƒVƒXƒeƒCƒ“‡¬y‘f‚Μ•‘‡‘ΜŒ`¬‹@\‚Μ‰π–Ύ‚Ζ”ρƒ^ƒ“ƒpƒN«ƒAƒ~ƒmŽ_ΆŽY‚Φ‚Μ‰ž—p kn-title=Analysis of a complex formation of cysteine synthase from Escherichia coli and its application to production of nonproteinaceous amino acids en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=ZhaoChunhui en-aut-sei=Zhao en-aut-mei=Chunhui kn-aut-name=ζβtτ kn-aut-sei=ζβ kn-aut-mei=tτ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw END start-ver=1.4 cd-journal=joma no-vol=1706 cd-vols= no-issue= article-no= start-page=464247 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230913 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Separation and fractionation of glutamic acid and histidine via origami isoelectric focusing en-subtitle= kn-subtitle= en-abstract= kn-abstract=We demonstrated the fractionation of two amino acids, glutamic acid and histidine, separated via isoelectric focusing (IEF) on filter paper folded and stacked in an origami fashion. Channels for electrophoresis were fabricated as circular zones acquired via wax printing onto the filter paper. An ampholyte solution with amphiphilic samples was deposited on all the circle zones, which was followed by folding to form the electrophoresis channels. IEF was achieved by applying an electrical potential between the anodic and cathodic chambers filled with phosphoric acid and sodium hydroxide solutions, respectively. A pH gradient was formed using either a wide-range ampholyte with a pH of 3 to 10 or a narrow-range version with a pH of 5 to 8, which was confirmed by adding pH indicators to each layer. The origami IEF was used to separate the amino acids, glutamic acid and histidine, by mixing with the ampholytes, which were deposited on the layers. The components in each layer were extracted with water and measured by high-performance liquid chromatography using pre-column derivatization with dansyl chloride. The results indicated that the focus for glutamic acid and that for histidine were at different layers, according to their isoelectric points. The origami isoelectric focusing achieved the fractionation of amino acids in less than 3 min using voltage as low as 30 V. en-copyright= kn-copyright= en-aut-name=DanchanaKaewta en-aut-sei=Danchana en-aut-mei=Kaewta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamashitaNayu en-aut-sei=Yamashita en-aut-mei=Nayu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=UmedaMika I. en-aut-sei=Umeda en-aut-mei=Mika I. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KanetaTakashi en-aut-sei=Kaneta en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Chemistry, Okayama University kn-affil= affil-num=2 en-affil=Department of Chemistry, Okayama University kn-affil= affil-num=3 en-affil=Department of Chemistry, Okayama University kn-affil= affil-num=4 en-affil=Department of Chemistry, Okayama University kn-affil= en-keyword=Paper-based analytical device kn-keyword=Paper-based analytical device en-keyword=Isoelectric focusing kn-keyword=Isoelectric focusing en-keyword=Origami electrophoresis kn-keyword=Origami electrophoresis en-keyword=Amino acids kn-keyword=Amino acids END start-ver=1.4 cd-journal=joma no-vol=89 cd-vols= no-issue=4 article-no= start-page=219 end-page=223 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230612 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Positive chemotaxis to plant apoplastic fluids of Pseudomonas syringae pv. tabaci 6605 and metabolome analysis en-subtitle= kn-subtitle= en-abstract= kn-abstract=Pseudomonas syringae pv. tabaci 6605 (Pta6605) is a causal agent of wildfire disease in host tobacco plants. Although chemotaxis has been shown to be necessary for Pta6605 in tobacco infection, the chemoattractants at the site of infection are unclear. Pta6605 was attracted to the apoplastic fluid from not only host tobacco leaves but also non-host plant leaves, indicating that Pta6605 is attracted to common plant metabolites. Metabolome analysis of apoplastic fluid from tobacco leaves revealed that amino acids including ƒΑ-aminobutyric acid and organic acids are abundant, suggesting that these compounds are potential chemoattractants. en-copyright= kn-copyright= en-aut-name=WatanabeYuta en-aut-sei=Watanabe en-aut-mei=Yuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TumewuStephany Angelia en-aut-sei=Tumewu en-aut-mei=Stephany Angelia kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamadaHajime en-aut-sei=Yamada en-aut-mei=Hajime kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MatsuiHidenori en-aut-sei=Matsui en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamamotoMikihiro en-aut-sei=Yamamoto en-aut-mei=Mikihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NoutoshiYoshiteru en-aut-sei=Noutoshi en-aut-mei=Yoshiteru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=ToyodaKazuhiro en-aut-sei=Toyoda en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=3 en-affil=Faculty of Agriculture, Okayama University kn-affil= affil-num=4 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=5 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=6 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=7 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=8 en-affil=The Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword=Apoplastic fluid kn-keyword=Apoplastic fluid en-keyword=Chemotaxis kn-keyword=Chemotaxis en-keyword=Chemoattractants kn-keyword=Chemoattractants en-keyword=Metabolome kn-keyword=Metabolome en-keyword=Pseudomonas syringae kn-keyword=Pseudomonas syringae END start-ver=1.4 cd-journal=joma no-vol=107 cd-vols= no-issue= article-no= start-page=52 end-page=59 dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=202304 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Comprehensive study of metabolic changes induced by a ketogenic diet therapy using GC/MS- and LC/MS-based metabolomics en-subtitle= kn-subtitle= en-abstract= kn-abstract=Objective: The ketogenic diet (KD), a high-fat and low-carbohydrate diet, is effective for a subset of patients with drug-resistant epilepsy, although the mechanisms of the KD have not been fully elucidated. The aims of this observational study were to investigate comprehensive short-term metabolic changes induced by the KD and to explore candidate metabolites or pathways for potential new therapeutic targets.
Methods: Subjects included patients with intractable epilepsy who had undergone the KD therapy (the medium-chain triglyceride [MCT] KD or the modified Atkins diet using MCT oil). Plasma and urine samples were obtained before and at 2?4 weeks after initiation of the KD. Targeted metabolome analyses of these samples were performed using gas chromatography-tandem mass spectrometry (GC/MS/MS) and liquid chromatography-tandem mass spectrometry (LC/MS/MS).
Results: Samples from 10 and 11 patients were analysed using GC/MS/MS and LC/MS/MS, respectively. The KD increased ketone bodies, various fatty acids, lipids, and their conjugates. In addition, levels of metabolites located upstream of acetyl-CoA and propionyl-CoA, including catabolites of branched-chain amino acids and structural analogues of ƒΑ-aminobutyric acid and lactic acid, were elevated.
Conclusions: The metabolites that were significantly changed after the initiation of the KD and related metabolites may be candidates for further studies for neuronal actions to develop new anti-seizure medications. en-copyright= kn-copyright= en-aut-name=AkiyamaMari en-aut-sei=Akiyama en-aut-mei=Mari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AkiyamaTomoyuki en-aut-sei=Akiyama en-aut-mei=Tomoyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SaigusaDaisuke en-aut-sei=Saigusa en-aut-mei=Daisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HishinumaEiji en-aut-sei=Hishinuma en-aut-mei=Eiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MatsukawaNaomi en-aut-sei=Matsukawa en-aut-mei=Naomi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=ShibataTakashi en-aut-sei=Shibata en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=TsuchiyaHiroki en-aut-sei=Tsuchiya en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MoriAtsushi en-aut-sei=Mori en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=FujiiYuji en-aut-sei=Fujii en-aut-mei=Yuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=MogamiYukiko en-aut-sei=Mogami en-aut-mei=Yukiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TokorodaniChiho en-aut-sei=Tokorodani en-aut-mei=Chiho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=KuwaharaKozue en-aut-sei=Kuwahara en-aut-mei=Kozue kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=Numata-UematsuYurika en-aut-sei=Numata-Uematsu en-aut-mei=Yurika kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=InoueKenji en-aut-sei=Inoue en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=KobayashiKatsuhiro en-aut-sei=Kobayashi en-aut-mei=Katsuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= affil-num=1 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= affil-num=2 en-affil=Department of Paediatrics (Child Neurology), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=4 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=5 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=6 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= affil-num=7 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= affil-num=8 en-affil=Department of Neurology, Shiga Medical Centre for Children kn-affil= affil-num=9 en-affil=Department of Paediatrics, Hiroshima City Funairi Citizens Hospital kn-affil= affil-num=10 en-affil=Department of Paediatric Neurology, Osaka Women's and Children's Hospital kn-affil= affil-num=11 en-affil=Department of Paediatrics, Kochi Health Sciences Centre kn-affil= affil-num=12 en-affil=Department of Paediatrics, Ehime Prefectural Central Hospital, kn-affil= affil-num=13 en-affil=Department of Paediatrics, Tohoku University School of Medicine kn-affil= affil-num=14 en-affil=Department of Neurology, Shiga Medical Centre for Children kn-affil= affil-num=15 en-affil=Department of Paediatrics (Child Neurology), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=Amino acids kn-keyword=Amino acids en-keyword=Biomarkers kn-keyword=Biomarkers en-keyword=Intractable epilepsy kn-keyword=Intractable epilepsy en-keyword=Ketone bodies kn-keyword=Ketone bodies en-keyword=Organic acids kn-keyword=Organic acids END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230324 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=ƒΐ-ƒqƒhƒƒLƒV-ƒΏ,ƒΏ-“ρ’uŠ· ƒΏ-ƒAƒ~ƒmŽ_\‘’‚Μ—§‘Μ‘I‘π\’z–@‚ΜŠJ”­ kn-title=Development of stereoselective constructions of ƒΐ-hydroxy-ƒΏ,ƒΏ-disubstituted ƒΏ-amino acid structures en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=ARAKIYuya en-aut-sei=ARAKI en-aut-mei=Yuya kn-aut-name=r–Ψ—Y–η kn-aut-sei=r–Ψ kn-aut-mei=—Y–η aut-affil-num=1 ORCID= affil-num=1 en-affil=Graduate School of Natural Science and Technology, Okayama university kn-affil=‰ͺŽR‘εŠw‘εŠw‰@Ž©‘R‰ΘŠwŒ€‹†‰Θ END start-ver=1.4 cd-journal=joma no-vol=15 cd-vols= no-issue=2 article-no= start-page=690 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230217 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Rationalizing the Binding Modes of PET Radiotracers Targeting the Norepinephrine Transporter en-subtitle= kn-subtitle= en-abstract= kn-abstract=Purpose: A new PET radiotracer F-18-AF78 showing great potential for clinical application has been reported recently. It belongs to a new generation of phenethylguanidine-based norepinephrine transporter (NET)-targeting radiotracers. Although many efforts have been made to develop NET inhibitors as antidepressants, systemic investigations of the structure-activity relationships (SARs) of NET-targeting radiotracers have rarely been performed. Methods: Without changing the phenethylguanidine pharmacophore and 3-fluoropropyl moiety that is crucial for easy labeling, six new analogs of F-18-AF78 with different meta-substituents on the benzene-ring were synthesized and evaluated in a competitive cellular uptake assay and in in vivo animal experiments in rats. Computational modeling of these tracers was established to quantitatively rationalize the interaction between the radiotracers and NET. Results: Using non-radiolabeled reference compounds, a competitive cellular uptake assay showed a decrease in NET-transporting affinity from meta-fluorine to iodine (0.42 and 6.51 mu M, respectively), with meta-OH being the least active (22.67 mu M). Furthermore, in vivo animal studies with radioisotopes showed that heart-to-blood ratios agreed with the cellular experiments, with AF78(F) exhibiting the highest cardiac uptake. This result correlates positively with the electronegativity rather than the atomic radius of the meta-substituent. Computational modeling studies revealed a crucial influence of halogen substituents on the radiotracer-NET interaction, whereby a T-shaped pi-pi stacking interaction between the benzene-ring of the tracer and the amino acid residues surrounding the NET binding site made major contributions to the different affinities, in accordance with the pharmacological data. Conclusion: The SARs were characterized by in vitro and in vivo evaluation, and computational modeling quantitatively rationalized the interaction between radiotracers and the NET binding site. These findings pave the way for further evaluation in different species and underline the potential of AF78(F) for clinical application, e.g., cardiac innervation imaging or molecular imaging of neuroendocrine tumors. en-copyright= kn-copyright= en-aut-name=TutovAnna en-aut-sei=Tutov en-aut-mei=Anna kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ChenXinyu en-aut-sei=Chen en-aut-mei=Xinyu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WernerRudolf A. en-aut-sei=Werner en-aut-mei=Rudolf A. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MuehligSaskia en-aut-sei=Muehlig en-aut-mei=Saskia kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ZimmermannThomas en-aut-sei=Zimmermann en-aut-mei=Thomas kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NoseNaoko en-aut-sei=Nose en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KoshinoKazuhiro en-aut-sei=Koshino en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=LapaConstantin en-aut-sei=Lapa en-aut-mei=Constantin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=DeckerMichael en-aut-sei=Decker en-aut-mei=Michael kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=HiguchiTakahiro en-aut-sei=Higuchi en-aut-mei=Takahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil=Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy and Food Chemistry, University of W?rzburg kn-affil= affil-num=2 en-affil=Nuclear Medicine, Faculty of Medicine, University of Augsburg kn-affil= affil-num=3 en-affil=Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital W?rzburg kn-affil= affil-num=4 en-affil=Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital W?rzburg kn-affil= affil-num=5 en-affil=Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy and Food Chemistry, University of W?rzburg kn-affil= affil-num=6 en-affil=Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=7 en-affil=Department of Systems and Informatics, Hokkaido Information University kn-affil= affil-num=8 en-affil=Nuclear Medicine, Faculty of Medicine, University of Augsburg kn-affil= affil-num=9 en-affil=Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy and Food Chemistry, University of W?rzburg kn-affil= affil-num=10 en-affil=Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=positron emission tomography kn-keyword=positron emission tomography en-keyword=norepinephrine transporter kn-keyword=norepinephrine transporter en-keyword=sympathetic nervous system kn-keyword=sympathetic nervous system en-keyword=structure-activity relationships kn-keyword=structure-activity relationships en-keyword=T-shaped ƒΞ?ƒΞ stacking kn-keyword=T-shaped ƒΞ?ƒΞ stacking END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue= article-no= start-page=1120710 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230223 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=E3-ubiquitin ligases and recent progress in osteoimmunology en-subtitle= kn-subtitle= en-abstract= kn-abstract=Ubiquitin-mediated proteasomal degradation is a post-transcriptional protein modification that is comprised of various components including the 76-amino acid protein ubiquitin (Ub), Ub-activating enzyme (E1), Ub-conjugating enzyme (E2), ubiquitin ligase (E3), deubiquitinating enzyme (DUB) and proteasome. We and others have recently provided genetic evidence showing that E3-ubiquitin ligases are associated with bone metabolism, the immune system and inflammation through ubiquitylation and subsequent degradation of their substrates. Dysregulation of the E3-ubiquitin ligase RNF146-mediated degradation of the adaptor protein 3BP2 (SH3 domain-binding protein 2) causes cherubism, an autosomal dominant disorder associated with severe inflammatory craniofacial dysmorphia syndrome in children. In this review, on the basis of our discoveries in cherubism, we summarize new insights into the roles of E3-ubiquitin ligases in the development of human disorders caused by an abnormal osteoimmune system by highlighting recent genetic evidence obtained in both human and animal model studies. en-copyright= kn-copyright= en-aut-name=AsanoYosuke en-aut-sei=Asano en-aut-mei=Yosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsumotoYoshinori en-aut-sei=Matsumoto en-aut-mei=Yoshinori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WadaJun en-aut-sei=Wada en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=RottapelRobert en-aut-sei=Rottapel en-aut-mei=Robert kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Princess Margaret Cancer Center, University Health Network, University of Toronto kn-affil= en-keyword=E3-ubiquitin ligases kn-keyword=E3-ubiquitin ligases en-keyword=ubiquitylation kn-keyword=ubiquitylation en-keyword=proteasomal degradation kn-keyword=proteasomal degradation en-keyword=osteoimmunology kn-keyword=osteoimmunology en-keyword=cherubism kn-keyword=cherubism END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue= article-no= start-page=e84291 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230228 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Chloride ions evoke taste sensations by binding to the extracellular ligand-binding domain of sweet/umami taste receptors en-subtitle= kn-subtitle= en-abstract= kn-abstract=Salt taste sensation is multifaceted: NaCl at low or high concentrations is preferably or aversively perceived through distinct pathways. Cl- is thought to participate in taste sensation through an unknown mechanism. Here, we describe Cl- ion binding and the response of taste receptor type 1 (T1r), a receptor family composing sweet/umami receptors. The T1r2a/T1r3 heterodimer from the medaka fish, currently the sole T1r amenable to structural analyses, exhibited a specific Cl- binding in the vicinity of the amino-acid-binding site in the ligand-binding domain (LBD) of T1r3, which is likely conserved across species, including human T1r3. The Cl- binding induced a conformational change in T1r2a/T1r3LBD at sub- to low-mM concentrations, similar to canonical taste substances. Furthermore, oral Cl- application to mice increased impulse frequencies of taste nerves connected to T1r-expressing taste cells and promoted their behavioral preferences attenuated by a T1r-specific blocker or T1r3 knock-out. These results suggest that the Cl- evokes taste sensations by binding to T1r, thereby serving as another preferred salt taste pathway at a low concentration. en-copyright= kn-copyright= en-aut-name=AtsumiNanako en-aut-sei=Atsumi en-aut-mei=Nanako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YasumatsuKeiko en-aut-sei=Yasumatsu en-aut-mei=Keiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TakashinaYuriko en-aut-sei=Takashina en-aut-mei=Yuriko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ItoChiaki en-aut-sei=Ito en-aut-mei=Chiaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MargolskeeRobert F. en-aut-sei=Margolskee en-aut-mei=Robert F. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=School of Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=6 en-affil=Monell Chemical Senses Center kn-affil= affil-num=7 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=taste receptor kn-keyword=taste receptor en-keyword=salt taste kn-keyword=salt taste en-keyword=chloride kn-keyword=chloride en-keyword=O kn-keyword=O en-keyword=latipes kn-keyword=latipes en-keyword=Mouse kn-keyword=Mouse en-keyword=Other kn-keyword=Other END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue=3 article-no= start-page=454 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2023 dt-pub=20230131 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The Pursuit of the "Inside" of the Amyloid Hypothesis-Is C99 a Promising Therapeutic Target for Alzheimer's Disease? en-subtitle= kn-subtitle= en-abstract= kn-abstract=Aducanumab, co-developed by Eisai (Japan) and Biogen (U.S.), has received Food and Drug Administration approval for treating Alzheimer's disease (AD). In addition, its successor antibody, lecanemab, has been approved. These antibodies target the aggregated form of the small peptide, amyloid-beta (A beta), which accumulates in the patient brain. The "amyloid hypothesis " based therapy that places the aggregation and toxicity of A beta at the center of the etiology is about to be realized. However, the effects of immunotherapy are still limited, suggesting the need to reconsider this hypothesis. A beta is produced from a type-I transmembrane protein, A beta precursor protein (APP). One of the APP metabolites, the 99-amino acids C-terminal fragment (C99, also called beta CTF), is a direct precursor of A beta and accumulates in the AD patient's brain to demonstrate toxicity independent of A beta. Conventional drug discovery strategies have focused on A beta toxicity on the "outside " of the neuron, but C99 accumulation might explain the toxicity on the "inside " of the neuron, which was overlooked in the hypothesis. Furthermore, the common region of C99 and A beta is a promising target for multifunctional AD drugs. This review aimed to outline the nature, metabolism, and impact of C99 on AD pathogenesis and discuss whether it could be a therapeutic target complementing the amyloid hypothesis. en-copyright= kn-copyright= en-aut-name=TakasugiNobumasa en-aut-sei=Takasugi en-aut-mei=Nobumasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KomaiMasato en-aut-sei=Komai en-aut-mei=Masato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KaneshiroNanaka en-aut-sei=Kaneshiro en-aut-mei=Nanaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=IkedaAtsuya en-aut-sei=Ikeda en-aut-mei=Atsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KamikuboYuji en-aut-sei=Kamikubo en-aut-mei=Yuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=UeharaTakashi en-aut-sei=Uehara en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Department of Medicinal Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Medicinal Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Division of Biomedical Sciences, School of Medicine, University of California kn-affil= affil-num=4 en-affil=Department of Medicinal Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Department of Cellular and Molecular Pharmacology, Juntendo University Graduate School of Medicine kn-affil= affil-num=6 en-affil=Department of Medicinal Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=Alzheimer's disease kn-keyword=Alzheimer's disease en-keyword=amyloid-beta kn-keyword=amyloid-beta en-keyword=amyloid beta precursor protein kn-keyword=amyloid beta precursor protein en-keyword=BACE1 kn-keyword=BACE1 en-keyword=C99 kn-keyword=C99 en-keyword=endolysosome kn-keyword=endolysosome en-keyword=autolysosome kn-keyword=autolysosome en-keyword=vesicular trafficking kn-keyword=vesicular trafficking END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue=50 article-no= start-page=46573 end-page=46582 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20221220 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Oligoarginine-Conjugated Peptide Foldamers Inhibiting Vitamin D Receptor-Mediated Transcription en-subtitle= kn-subtitle= en-abstract= kn-abstract=The vitamin D receptor (VDR) is a nuclear receptor, which is involved in several physiological processes, including differentiation and bone homeostasis. The VDR is a promising target for the development of drugs against cancer and bone-related diseases. To date, several VDR antagonists, which bind to the ligand binding domain of the VDR and compete with the endogenous agonist 1 alpha,25(OH)D3, have been reported. However, these ligands contain a secosteroidal skeleton, which is chemically unstable and complicated to synthesize. A few VDR antagonists with a nonsecosteroidal skeleton have been reported. Alternative inhibitors against VDR transactivation that act via different mechanisms are desirable. Here, we developed peptide-based VDR inhibitors capable of disrupting the VDR-coactivator interaction. It was reported that helical SRC2-3 peptides strongly bound to the VDR and competed with the coactivator in vitro. Therefore, we designed and synthesized a series of SRC2-3 derivatives by the introduction of nonproteinogenic amino acids, such as beta-amino acids, and by side-chain stapling to stabilize helical structures and provide resistance against digestive enzymes. In addition, conjugation with a cell-penetrating peptide increased the cell membrane permeability and was a promising strategy for intracellular VDR inhibition. The nona-arginine-conjugated peptides 24 with side-chain stapling and 25 with cyclic beta-amino acids showed strong intracellular VDR inhibitory activity, resulting in suppression of the target gene expression and inhibition of the cell differentiation of HL-60 cells. Herein, the peptide design, structure-activity relationship (SAR) study, and biological evaluation of the peptides are described. en-copyright= kn-copyright= en-aut-name=TakyoMami en-aut-sei=Takyo en-aut-mei=Mami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SatoYumi en-aut-sei=Sato en-aut-mei=Yumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HirataNaoya en-aut-sei=Hirata en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TsuchiyaKeisuke en-aut-sei=Tsuchiya en-aut-mei=Keisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=IshidaHiroaki en-aut-sei=Ishida en-aut-mei=Hiroaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KuroharaTakashi en-aut-sei=Kurohara en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YanaseYuta en-aut-sei=Yanase en-aut-mei=Yuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=ItoTakahito en-aut-sei=Ito en-aut-mei=Takahito kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KandaYasunari en-aut-sei=Kanda en-aut-mei=Yasunari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=YamamotoKeiko en-aut-sei=Yamamoto en-aut-mei=Keiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=MisawaTakashi en-aut-sei=Misawa en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=DemizuYosuke en-aut-sei=Demizu en-aut-mei=Yosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= affil-num=1 en-affil=National Institute of Health Sciences kn-affil= affil-num=2 en-affil=National Institute of Health Sciences kn-affil= affil-num=3 en-affil=National Institute of Health Sciences kn-affil= affil-num=4 en-affil=National Institute of Health Sciences kn-affil= affil-num=5 en-affil=Laboratory of Drug Design and Medicinal Chemistry, Showa Pharmaceutical University kn-affil= affil-num=6 en-affil=National Institute of Health Sciences kn-affil= affil-num=7 en-affil=National Institute of Health Sciences kn-affil= affil-num=8 en-affil=National Institute of Health Sciences kn-affil= affil-num=9 en-affil=National Institute of Health Sciences kn-affil= affil-num=10 en-affil=Laboratory of Drug Design and Medicinal Chemistry, Showa Pharmaceutical University kn-affil= affil-num=11 en-affil=National Institute of Health Sciences kn-affil= affil-num=12 en-affil=National Institute of Health Sciences kn-affil= END start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=6 article-no= start-page=654 end-page=665 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202212 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Neuropeptide Y Antagonizes Development of Pulmonary Fibrosis through IL-1ƒΐ Inhibition en-subtitle= kn-subtitle= en-abstract= kn-abstract=Neuropeptide Y (NPY), a 36 amino acid residue polypeptide distributed throughout the nervous system, acts on various immune cells in many organs, including the respiratory system. However, little is known about its role in the pathogenesis of pulmonary fibrosis. This study was performed to determine the effects of NPY on pulmonary fibrosis. NPY-deficient and wild-type mice were intratracheally administered bleomycin. Inflammatory cells, cytokine concentrations, and morphological morphometry of the lungs were analyzed. Serum NPY concentrations were also measured in patients with idiopathic pulmonary fibrosis and healthy control subjects. NPY-deficient mice exhibited significantly enhanced pulmonary fibrosis and higher IL-1 beta concentrations in the lungs compared with wild-type mice. Exogenous NPY treatment suppressed the development of bleomycin-induced lung fibrosis and decreased IL-1 beta concentrations in the lungs. Moreover, IL-1 beta neutralization in NPY-deficient mice attenuated the fibrotic changes. NPY decreased IL-1 beta release, and Y1 receptor antagonists inhibited IL-1 beta release and induced epithelial-mesenchymal transition in human alveolar epithelial cells. Patients with idiopathic pulmonary fibrosis had lower NPY and greater IL-1 beta concentrations in the serums compared with healthy control subjects. NPY expression was mainly observed around bronchial epithelial cells in human idiopathic pulmonary fibrosis lungs. These data suggest that NPY plays a protective role against pulmonary fibrosis by suppressing IL-1 beta release, and manipulating the NPY-Y1 receptor axis could be a potential therapeutic strategy for delaying disease progression. en-copyright= kn-copyright= en-aut-name=ItanoJunko en-aut-sei=Itano en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TaniguchiAkihiko en-aut-sei=Taniguchi en-aut-mei=Akihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SenooSatoru en-aut-sei=Senoo en-aut-mei=Satoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=AsadaNoboru en-aut-sei=Asada en-aut-mei=Noboru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=GionYuka en-aut-sei=Gion en-aut-mei=Yuka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=EgusaYuria en-aut-sei=Egusa en-aut-mei=Yuria kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=GuoLili en-aut-sei=Guo en-aut-mei=Lili kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=OdaNaohiro en-aut-sei=Oda en-aut-mei=Naohiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=ArakiKota en-aut-sei=Araki en-aut-mei=Kota kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=SatoYasuharu en-aut-sei=Sato en-aut-mei=Yasuharu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=ToyookaShinichi en-aut-sei=Toyooka en-aut-mei=Shinichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=KiuraKatsuyuki en-aut-sei=Kiura en-aut-mei=Katsuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=MaedaYoshinobu en-aut-sei=Maeda en-aut-mei=Yoshinobu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=MiyaharaNobuaki en-aut-sei=Miyahara en-aut-mei=Nobuaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= affil-num=1 en-affil=Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Hematology and Oncology, Okayama University Hospital kn-affil= affil-num=5 en-affil=Department of Medical Technology, Okayama University Graduate School of Health Sciences kn-affil= affil-num=6 en-affil=Department of Medical Technology, Okayama University Graduate School of Health Sciences kn-affil= affil-num=7 en-affil=Department of Medical Technology, Okayama University Graduate School of Health Sciences kn-affil= affil-num=8 en-affil=Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=9 en-affil=Department of General Thoracic Surgery, Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=10 en-affil=Department of Medical Technology, Okayama University Graduate School of Health Sciences kn-affil= affil-num=11 en-affil=Department of General Thoracic Surgery, Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=12 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= affil-num=13 en-affil=Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=14 en-affil=Department of Allergy and Respiratory Medicine, Okayama University Hospital kn-affil= en-keyword=idiopathic pulmonary fibrosis kn-keyword=idiopathic pulmonary fibrosis en-keyword=NPY kn-keyword=NPY en-keyword=IL-1 beta; bleomycin kn-keyword=IL-1 beta; bleomycin en-keyword=bronchial epithelial cells kn-keyword=bronchial epithelial cells END start-ver=1.4 cd-journal=joma no-vol=471 cd-vols= no-issue= article-no= start-page=214742 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202211 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Geometric, electronic and spin structures of the CaMn4O5 catalyst for water oxidation in oxygen-evolving photosystem II. Interplay between experiments and theoretical computations en-subtitle= kn-subtitle= en-abstract= kn-abstract=The aim of this review is to elucidate geometric structures of the catalytic CaMn4Ox (x = 5, 6) cluster in the Kok cycle for water oxidation in the oxygen evolving complex (OEC) of photosystem II (PSII) based on the high-resolution (HR) X-ray diffraction (XRD) and serial femtosecond crystallography (SFX) experiments using the X-ray free-electron laser (XFEL). Quantum mechanics (QM) and QM/molecular mechanics (MM) computations are performed to elucidate the electronic and spin structures of the CaMn4Ox (x = 5, 6) cluster in five states S-i (i = 0 similar to 4) on the basis of the X-ray spectroscopy, electron paramagnetic resonance (EPR) and related experiments. Interplay between the experiments and theoretical computations has been effective to elucidate the coordination structures of the CaMn4Ox (x = 5, 6) cluster ligated by amino acid residues of the protein matrix of PSII, valence states of the four Mn ions and total spin states by their exchange-couplings, and proton-shifted isomers of the CaMn4Ox (x = 5, 6) cluster. The HR XRD and SFX XFEL experiments have also elucidated the biomolecular systems structure of OEC of PSII and the hydrogen bonding networks consisting of water molecules, chloride anions, etc., for water inlet and proton release pathways in PSII. Large-scale QM/MM computations have been performed for elucidation of the hydrogen bonding distances and angles by adding invisible hydrogen atoms to the HR XRD structure. Full geometry optimizations by the QM and QM/MM methods have been effective for elucidation of the molecular systems structure around the CaMn4Ox (x = 5, 6) cluster in OEC. DLPNO-CCSD(T-0) method has been applied to elucidate relative energies of possible intermediates in each state of the Kok cycle for water oxidation. Implications of these results are discussed in relation to the blueprint for developments of artificial catalysts for water oxidation. en-copyright= kn-copyright= en-aut-name=YamaguchiKizashi en-aut-sei=Yamaguchi en-aut-mei=Kizashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ShojiMitsuo en-aut-sei=Shoji en-aut-mei=Mitsuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IsobeHiroshi en-aut-sei=Isobe en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KawakamiTakashi en-aut-sei=Kawakami en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MiyagawaKoichi en-aut-sei=Miyagawa en-aut-mei=Koichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SugaMichihiro en-aut-sei=Suga en-aut-mei=Michihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=AkitaFusamichi en-aut-sei=Akita en-aut-mei=Fusamichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=ShenJian-Ren en-aut-sei=Shen en-aut-mei=Jian-Ren kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Center for Quantum Information and Quantum Biology, Osaka University kn-affil= affil-num=2 en-affil=Center of Computational Sciences, Tsukuba University kn-affil= affil-num=3 en-affil=Research Institute for Interdisciplinary Science, and Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=4 en-affil=RIKEN Center for Computational Science kn-affil= affil-num=5 en-affil=Center of Computational Sciences, Tsukuba University kn-affil= affil-num=6 en-affil=Research Institute for Interdisciplinary Science, and Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=7 en-affil=Research Institute for Interdisciplinary Science, and Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=8 en-affil=Research Institute for Interdisciplinary Science, and Graduate School of Natural Science and Technology, Okayama University kn-affil= en-keyword=Water oxidation kn-keyword=Water oxidation en-keyword=Oxygen evolution kn-keyword=Oxygen evolution en-keyword=Photosystem II kn-keyword=Photosystem II en-keyword=HR XRD kn-keyword=HR XRD en-keyword=SFX XFEL kn-keyword=SFX XFEL en-keyword=QM/MM calculation kn-keyword=QM/MM calculation en-keyword=DLPNO CCSD(T-0) computations, Oxyl radical character kn-keyword=DLPNO CCSD(T-0) computations, Oxyl radical character END start-ver=1.4 cd-journal=joma no-vol=38 cd-vols= no-issue=12 article-no= start-page=241 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20221022 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Second extracellular protease mediating maturation of Vibrio mimicus?hemolysin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Vibrio mimicus is a bacterium that causes gastroenteritis in humans. This pathogen produces an enterotoxic hemolysin called V. mimicus hemolysin (VMH), which is secreted extracellularly as an inactive 80-kDa protoxin and converted to a 66-kDa mature toxin through cleavage between Arg(151) and Ser(152). The 56-kDa serine protease termed V. mimicus trypsin-like protease (VmtA) is known to mediate this maturating process. However, some strains including strain ES-20 does not possess the vmtA gene. In the present study, the vmtA-negative strains were found to have a replaced gene that encodes a 43-kDa (403 aa) precursor of a serine protease designated by VmtX (V. mimicus trypsin-like protease X). To examine whether VmtX is also involved in the maturation of VMH, VmtX was isolated from the culture supernatant of V. mimicus strain NRE-20, a metalloprotease-negative mutant constructed from strain ES-20. Concretely, the culture supernatant was fractionated with 70% saturated ammonium sulfate and subjected to affinity column chromatography using a HiTrap Benzamidine FF column. The analysis of the N-terminal amino acid sequences of the proteins in the obtained VmtX preparation indicated that the 39-kDa protein was active VmtX consisting of 371 aa (Ile(33)-Ser(403)). The VmtX preparation was found to activate pro-VMH through generation of the 66-kDa protein. Additionally, treatment of the VmtX preparation with serine protease inhibitors, such as leupeptin and phenylmethylsulfonyl fluoride, significantly suppressed the activities to hydrolyze the specific peptide substrate and to synthesize the 66-kDa toxin. These findings indicate that VmtX is the second protease that mediats the maturation of VMH. en-copyright= kn-copyright= en-aut-name=MiyoshiShin-ichi en-aut-sei=Miyoshi en-aut-mei=Shin-ichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TokoNorie en-aut-sei=Toko en-aut-mei=Norie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=DodoTetsuya en-aut-sei=Dodo en-aut-mei=Tetsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=NankoAyako en-aut-sei=Nanko en-aut-mei=Ayako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MizunoTamaki en-aut-sei=Mizuno en-aut-mei=Tamaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=Vibrio mimicus kn-keyword=Vibrio mimicus en-keyword=Serine protease kn-keyword=Serine protease en-keyword=Hemolysin kn-keyword=Hemolysin en-keyword=Maturation kn-keyword=Maturation END start-ver=1.4 cd-journal=joma no-vol=11 cd-vols= no-issue=9 article-no= start-page=1805 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202209 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Rice Nudix Hydrolase OsNUDX2 Sanitizes Oxidized Nucleotides en-subtitle= kn-subtitle= en-abstract= kn-abstract=Nudix hydrolase (NUDX) hydrolyzes 8-oxo-(d)GTP to reduce the levels of oxidized nucleotides in the cells. 8-oxo-(d)GTP produced by reactive oxygen species (ROS) is incorporated into DNA/RNA and mispaired with adenine, causing replicational and transcriptional errors. Here, we identified a rice OsNUDX2 gene, whose expression level was increased 15-fold under UV-C irradiation. The open reading frame of the OsNUDX2 gene, which encodes 776 amino acid residues, was cloned into Escherichia coli cells to produce the protein of 100 kDa. The recombinant protein hydrolyzed 8-oxo-dGTP, in addition to dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP), as did Arabidopsis AtNUDX1; whereas the amino acid sequence of OsNUDX2 had 18% identity with AtNUDX1. OsNUDX2 had 14% identity with barley HvNUDX12, which hydrolyzes 8-oxo-dGTP and diadenosine tetraphosphates. Suppression of the lacZ amber mutation caused by the incorporation of 8-oxo-GTP into mRNA was prevented to a significant degree when the OsNUDX2 gene was expressed in mutT-deficient E. coli cells. These results suggest that the different substrate specificity and identity among plant 8-oxo-dGTP-hydrolyzing NUDXs and OsNUDX2 reduces UV stress by sanitizing the oxidized nucleotides. en-copyright= kn-copyright= en-aut-name=KondoYuki en-aut-sei=Kondo en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=RikiishiKazuhide en-aut-sei=Rikiishi en-aut-mei=Kazuhide kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SugimotoManabu en-aut-sei=Sugimoto en-aut-mei=Manabu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=8-oxo-dGTP kn-keyword=8-oxo-dGTP en-keyword=nudix hydrolase kn-keyword=nudix hydrolase en-keyword=Oryza sativa kn-keyword=Oryza sativa en-keyword=transcriptional error kn-keyword=transcriptional error en-keyword=UV-C kn-keyword=UV-C END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=17 article-no= start-page=3541 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220827 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Dose-Dependent Effects of Amino Acids on Clinical Outcomes in Adult Medical Inpatients Receiving Only Parenteral Nutrition: A Retrospective Cohort Study Using a Japanese Medical Claims Database en-subtitle= kn-subtitle= en-abstract= kn-abstract=The majority of inpatients requiring parenteral nutrition (PN) do not receive adequate amino acid, which may negatively impact clinical outcomes. We investigated the influence of amino acid doses on clinical outcomes in medical adult inpatients fasting >10 days and receiving only PN, using Japanese medical claims database. The primary endpoint was in-hospital mortality, and the secondary endpoints included deterioration of activities of daily living (ADL), intravenous catheter infection, hospital readmission, hospital length of stay (LOS), and total medical costs. Patients were divided into four groups according to their mean prescribed daily amino acid doses from Days 4 to 10 of fasting: Adequate (>= 0.8 g/kg/day), Moderate (>= 0.6-<0.8 g/kg/day), Low (>= 0.4-<0.6 g/kg/day), and Very low (<0.4 g/kg/day). Multivariate logistic or multiple regression analyses were performed with adjustments for patient characteristics (total n = 86,702). The Adequate group was used as the reference in all analyses. For the Moderate, Low, and Very low groups, adjusted ORs (95% CI) of in-hospital mortality were 1.20 (1.14-1.26), 1.43 (1.36-1.51), and 1.72 (1.62-1.82), respectively, and for deterioration of ADL were 1.21 (1.11-1.32), 1.34 (1.22-1.47), and 1.22 (1.09-1.37), respectively. Adjusted regression coefficients (95% CI) of hospital LOS were 1.2 (0.4-2.1), 1.5 (0.6-2.4), and 2.9 (1.8-4.1), respectively. Lower prescribed doses of amino acids were associated with worse clinical outcomes including higher in-hospital mortality. en-copyright= kn-copyright= en-aut-name=TakagiKosei en-aut-sei=Takagi en-aut-mei=Kosei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MurotaniKenta en-aut-sei=Murotani en-aut-mei=Kenta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KamoshitaSatoru en-aut-sei=Kamoshita en-aut-mei=Satoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KurodaAkiyoshi en-aut-sei=Kuroda en-aut-mei=Akiyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, kn-affil= affil-num=2 en-affil=Biostatistics Center, Kurume University kn-affil= affil-num=3 en-affil=Medical Affairs Department, Research and Development Center, Otsuka Pharmaceutical Factory, Inc. kn-affil= affil-num=4 en-affil=Research and Development Center, Otsuka Pharmaceutical Factory, Inc. kn-affil= en-keyword=parenteral nutrition kn-keyword=parenteral nutrition en-keyword=amino acids kn-keyword=amino acids en-keyword=medical inpatient kn-keyword=medical inpatient en-keyword=clinical outcomes kn-keyword=clinical outcomes en-keyword=real-world data kn-keyword=real-world data END start-ver=1.4 cd-journal=joma no-vol=76 cd-vols= no-issue=4 article-no= start-page=423 end-page=428 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202208 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effects of Exercise Therapy and Nutrition Therapy on Patients with Possible Malnutrition and Sarcopenia in a Recovery Rehabilitation Ward en-subtitle= kn-subtitle= en-abstract= kn-abstract=We compared the effects of an exercise intervention with that of exercise combined with nutrition therapy in patients with possible malnutrition and sarcopenia admitted to a recovery rehabilitation ward, and we examined the differences in the patientsf physical function and activities of daily living (ADLs). There were 16 patients in the Exercise group with exercise therapy and ADL exercises, and 14 patients in the Combined intervention group with exercise therapy, ADL exercises, and nutrition therapy. The survey items were body weight, body mass index, grip strength, lower-leg circumference, gait speed, and ADLs, each of which was measured at the baseline and at 2 weeks, 4 weeks, and at discharge. Significant improvements in grip strength were observed in the Combined intervention group as follows: at 4 weeks>at 2 weeks (p<0.05), and at discharge>baseline and 2 weeks (p<0.05). There were no significant changes in the Exercise group, and an interaction was recognized in both groups. Comprehensive rehabilitation including nutrition therapy is necessary for patients with possible malnutrition and/or sarcopenia, as our results indicate that nutrition therapy in addition to exercise therapy has the effect of promoting improvements of physical function in such patients. en-copyright= kn-copyright= en-aut-name=TakahashiSatoshi en-aut-sei=Takahashi en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KushibeTakuya en-aut-sei=Kushibe en-aut-mei=Takuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AkezakiYoshiteru en-aut-sei=Akezaki en-aut-mei=Yoshiteru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HoriikeNorio en-aut-sei=Horiike en-aut-mei=Norio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Rehabilitation Medicine, Saiseikai Imabari Daini Hospital kn-affil= affil-num=2 en-affil=Department of Rehabilitation Medicine, Saiseikai Imabari Daini Hospital kn-affil= affil-num=3 en-affil=Division of Physical Therapy, Kochi Professional University of Rehabilitation kn-affil= affil-num=4 en-affil=Department of Internal Medicine, Saiseikai Imabari Daini Hospital kn-affil= en-keyword=sarcopenia kn-keyword=sarcopenia en-keyword=rehabilitation kn-keyword=rehabilitation en-keyword=exercise therapy kn-keyword=exercise therapy en-keyword=nutrition therapy kn-keyword=nutrition therapy en-keyword=grip strength kn-keyword=grip strength END start-ver=1.4 cd-journal=joma no-vol=76 cd-vols= no-issue=4 article-no= start-page=415 end-page=421 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202208 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=5-Nitro-2-(3-phenylpropylamino) Benzoic Acid Inhibits the Proliferation and Migration of Lens Epithelial Cells by Blocking CaMKII Signaling en-subtitle= kn-subtitle= en-abstract= kn-abstract=Posterior capsule opacification (PCO) is a post-surgery complication of cataract surgery, and lens epithelial cells (LECs) are involved in its development. A suppressive effect on LECs is exerted by the non specific chloride channel inhibitor 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) exerts. Herein, the growth and migration inhibitory effects of NPPB on LECs were assessed, and the mechanism underlying the effects were investigated by focusing on Ca2+/CaMKII signaling. LECs were treated with different concentrations of NPPB, and the changes in cell viability, cell-cycle distribution, anchorage-dependent growth, migration, Ca2+ level, and CaMKII expression were evaluated. NPPB inhibited LECsf proliferation and induced G1 cell-cycle arrest in the cells. Regarding LECsf mobility, NPPB suppressed the cellsf anchorage-dependent growth ability and inhibited their migration. Changes in cell phenotypes were associated with an increased intracellular Ca2+ level and down-regulation of CaMKII. Together these results confirmed the inhibitory effect of NPPB on the proliferation and migration of LECs, and the effect was shown to be associated with the induced level of Ca2+ and the inhibition of CaMKII signaling transduction. en-copyright= kn-copyright= en-aut-name=KangHaijun en-aut-sei=Kang en-aut-mei=Haijun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HuangDongmei en-aut-sei=Huang en-aut-mei=Dongmei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KangGangjin en-aut-sei=Kang en-aut-mei=Gangjin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YangXu en-aut-sei=Yang en-aut-mei=Xu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=LiHeng en-aut-sei=Li en-aut-mei=Heng kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=LiuSiyuan en-aut-sei=Liu en-aut-mei=Siyuan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=GouWenjun en-aut-sei=Gou en-aut-mei=Wenjun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=LiuLinglin en-aut-sei=Liu en-aut-mei=Linglin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=QiuYuyan en-aut-sei=Qiu en-aut-mei=Yuyan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=2 en-affil=Department of Cardiovascular, Suining Central Hospital kn-affil= affil-num=3 en-affil=Department of Ophthalmology, The Affiliated Hospital of Southwest Medical University kn-affil= affil-num=4 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=5 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=6 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=7 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=8 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= affil-num=9 en-affil=Department of Ophthalmology, Suining Central Hospital kn-affil= en-keyword=5-nitro-2-(3-phenylpropylamino) benzoic acid kn-keyword=5-nitro-2-(3-phenylpropylamino) benzoic acid en-keyword=CaMKII kn-keyword=CaMKII en-keyword=lens epithelial cell kn-keyword=lens epithelial cell en-keyword=migration kn-keyword=migration en-keyword=proliferation kn-keyword=proliferation END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=8 article-no= start-page=1722 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220804 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A Transfectable Fusagravirus from a Japanese Strain of Cryphonectria carpinicola with Spherical Particles en-subtitle= kn-subtitle= en-abstract= kn-abstract=A novel dsRNA virus (Cryphonectria carpinicola fusagravirus 1, CcFGV1), isolated from a Japanese strain (JS13) of Cryphonectria carpinicola, was thoroughly characterized. The biological comparison of a set of isogenic CcFGV1-infected and -free (JS13VF) strains indicated asymptomatic infection by CcFGV1. The sequence analysis showed that the virus has a two open reading frame (ORF) genome of 9.6 kbp with the RNA-directed RNA polymerase domain encoded by ORF2. The N-terminal sequencing and peptide mass fingerprinting showed an N-terminally processed or degraded product (150 kDa) of the 5'-proximal ORF1-encoded protein (1462 amino acids) to make up the CcFGV1 spherical particles of similar to 40 nm in diameter. Interestingly, a portion of CcFGV1 dsRNA co-fractionated with a host protein of 70 kDa. The purified CcFGV1 particles were used to transfect protoplasts of JS13VF as well as the standard strain of an experimental model filamentous fungal host Cryphonectria parasitica. CcFGV1 was confirmed to be associated with asymptomatic infection of both fungi. RNA silencing was shown to target the virus in C. parasitica, resulting in reduced CcFGV1 accumulation by comparing the CcFGV1 content between RNA silencing-competent and -deficient strains. These results indicate the transfectability of spherical particles of a fusagravirus associated with asymptomatic infection. en-copyright= kn-copyright= en-aut-name=DasSubha en-aut-sei=Das en-aut-mei=Subha kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HisanoSakae en-aut-sei=Hisano en-aut-mei=Sakae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=Eusebio-CopeAna en-aut-sei=Eusebio-Cope en-aut-mei=Ana kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KondoHideki en-aut-sei=Kondo en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SuzukiNobuhiro en-aut-sei=Suzuki en-aut-mei=Nobuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=4 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=5 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=Cryphonectria carpinicola kn-keyword=Cryphonectria carpinicola en-keyword=Cryphonectria parasitica kn-keyword=Cryphonectria parasitica en-keyword=fusagravirus kn-keyword=fusagravirus en-keyword=fungal virus kn-keyword=fungal virus en-keyword=dsRNA kn-keyword=dsRNA en-keyword=spherical virion kn-keyword=spherical virion en-keyword=transfection kn-keyword=transfection END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue=1 article-no= start-page=13540 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220808 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=RNA editing facilitates the enhanced production of neoantigens during the simultaneous administration of oxaliplatin and radiotherapy in colorectal cancer en-subtitle= kn-subtitle= en-abstract= kn-abstract=Most cases of colorectal cancers (CRCs) are microsatellite stable (MSS), which frequently demonstrate lower response rates to immune checkpoint inhibitors (ICIs). RNA editing produces neoantigens by altering amino acid sequences. In this study, RNA editing was induced artificially by chemoradiation therapy (CRT) to generate neoantigens in MSS CRCs. Altogether, 543 CRC specimens were systematically analyzed, and the expression pattern of ADAR1 was investigated. In vitro and in vivo experiments were also performed. The RNA editing enzyme ADAR1 was upregulated in microsatellite instability-high CRCs, leading to their high affinity for ICIs. Although ADAR1 expression was low in MSS CRC, CRT including oxaliplatin (OX) treatment upregulated RNA editing levels by inducing ADAR1. Immunohistochemistry analyses showed the upregulation of ADAR1 in patients with CRC treated with CAPDX (capecitabine +OX) radiation therapy relative to ADAR1 expression in patients with CRC treated only by surgery (p <0.001). Compared with other regimens, CRT with OX effectively induced RNA editing in MSS CRC cell lines (HT29 and Caco2, p <0.001) via the induction of type 1 interferon-triggered ADAR1 expression. CRT with OX promoted the RNA editing of cyclin I, a neoantigen candidate. Neoantigens can be artificially induced by RNA editing via an OX-CRT regimen. CRT can promote proteomic diversity via RNA editing. en-copyright= kn-copyright= en-aut-name=KomatsuYasuhiro en-aut-sei=Komatsu en-aut-mei=Yasuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ShigeyasuKunitoshi en-aut-sei=Shigeyasu en-aut-mei=Kunitoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YanoShuya en-aut-sei=Yano en-aut-mei=Shuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakedaSho en-aut-sei=Takeda en-aut-mei=Sho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TakahashiKazutaka en-aut-sei=Takahashi en-aut-mei=Kazutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HataNanako en-aut-sei=Hata en-aut-mei=Nanako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=UmedaHibiki en-aut-sei=Umeda en-aut-mei=Hibiki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=YoshidaKazuhiro en-aut-sei=Yoshida en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=MoriYoshiko en-aut-sei=Mori en-aut-mei=Yoshiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=YasuiKazuya en-aut-sei=Yasui en-aut-mei=Kazuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=YoshidaRyuichi en-aut-sei=Yoshida en-aut-mei=Ryuichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=KondoYoshitaka en-aut-sei=Kondo en-aut-mei=Yoshitaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=KishimotoHiroyuki en-aut-sei=Kishimoto en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=TeraishiFuminori en-aut-sei=Teraishi en-aut-mei=Fuminori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=UmedaYuzo en-aut-sei=Umeda en-aut-mei=Yuzo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= en-aut-name=KagawaShunsuke en-aut-sei=Kagawa en-aut-mei=Shunsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=16 ORCID= en-aut-name=MichiueHiroyuki en-aut-sei=Michiue en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=17 ORCID= en-aut-name=TazawaHiroshi en-aut-sei=Tazawa en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=18 ORCID= en-aut-name=GoelAjay en-aut-sei=Goel en-aut-mei=Ajay kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=19 ORCID= en-aut-name=FujiwaraToshiyoshi en-aut-sei=Fujiwara en-aut-mei=Toshiyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=20 ORCID= affil-num=1 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=9 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=10 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=11 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=12 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=13 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=14 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=15 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=16 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=17 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=18 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= affil-num=19 en-affil=Department of Molecular Diagnostics and Experimental Therapeutics, Beckman Research Institute, City of Hope Biomedical Research Center kn-affil= affil-num=20 en-affil=Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= END start-ver=1.4 cd-journal=joma no-vol=76 cd-vols= no-issue=3 article-no= start-page=255 end-page=263 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202206 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Intrathecal Administration of the ƒΏ1 Adrenergic Antagonist Phentolamine Upregulates Spinal GLT-1 and Improves Mirror Image Pain in SNI Model Rats en-subtitle= kn-subtitle= en-abstract= kn-abstract=Mirror image pain (MIP) is a type of extraterritorial pain that results in contralateral pain or allodynia. Glutamate transporter-1 (GLT-1) is expressed in astrocytes and plays a role in maintaining low glutamate levels in the synaptic cleft. Previous studies have shown that GLT-1 dysfunction induces neuropathic pain. Our previous study revealed bilateral GLT-1 downregulation in the spinal cord of a spared nerve injury (SNI) rat. We hypothesized that spinal GLT-1 is involved in the mechanism of MIP. We also previously demonstrated noradrenergic GLT-1 regulation. Therefore, this study aimed to investigate the effect of an ƒΏ1 adrenergic antagonist on the development of MIP. Rats were subjected to SNI. Changes in pain behavior and GLT-1 protein levels in the SNI rat spinal cords were then examined by intrathecal administration of the ƒΏ1 adrenergic antagonist phentolamine, followed by von Frey test and western blotting. SNI resulted in the development of MIP and bilateral downregulation of GLT-1 protein in the rat spinal cord. Intrathecal phentolamine increased contralateral GLT-1 protein levels and partially ameliorated the 50% paw withdrawal threshold in the contralateral hind paw. Spinal GLT-1 upregulation by intrathecal phentolamine ameliorates MIP. GLT-1 plays a role in the development of MIPs. en-copyright= kn-copyright= en-aut-name=NakatsukaKosuke en-aut-sei=Nakatsuka en-aut-mei=Kosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsuokaYoshikazu en-aut-sei=Matsuoka en-aut-mei=Yoshikazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KuritaMasako en-aut-sei=Kurita en-aut-mei=Masako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=WangRuilin en-aut-sei=Wang en-aut-mei=Ruilin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TsuboiChika en-aut-sei=Tsuboi en-aut-mei=Chika kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SueNobutaka en-aut-sei=Sue en-aut-mei=Nobutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KakuRyuji en-aut-sei=Kaku en-aut-mei=Ryuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MorimatsuHiroshi en-aut-sei=Morimatsu en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Kinoshita Pain Clinic kn-affil= affil-num=4 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=alpha adrenergic receptor kn-keyword=alpha adrenergic receptor en-keyword=glutamate transporter-1 kn-keyword=glutamate transporter-1 en-keyword=mirror image pain kn-keyword=mirror image pain en-keyword=neuropathic pain kn-keyword=neuropathic pain en-keyword=spared nerve injury kn-keyword=spared nerve injury END start-ver=1.4 cd-journal=joma no-vol=76 cd-vols= no-issue=3 article-no= start-page=235 end-page=245 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=202206 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Roles of Transmembrane Protein 97 (TMEM97) in Adipose Tissue and Skeletal Muscle en-subtitle= kn-subtitle= en-abstract= kn-abstract=The combination of sarcopenia and obesity (sarcopenic obesity) is associated with the development of metabolic syndrome and cardiovascular events. The molecular pathways that develop sarcopenic obesity have studied intensively. Transmembrane protein 97 (TMEM97) is 176 amino acids conserved integral membrane protein with four transmembrane domains that is expressed in several types of cancer. Its physiological significance in adipose tissue and skeletal muscle has been unclear. We studied TMEM97-transgenic mice and mice lacking TMEM97, and our findings indicate that TMEM97 expression is regulated in adipose tissue and skeletal muscle from obesity. TMEM97 represses adipogenesis and promotes myogenesis in vitro. Fat-specific TMEM97 transgenic mice showed systemic insulin resistance. Mice overexpressing TMEM97 in skeletal muscle exhibited systemic insulin resistance. Mice lacking TMEM97 were protected against diet-induced obesity and insulin resistance. These phenotypes are associated with the effects of TMEM97 on inflammation genes in adipose tissue and skeletal muscle. Our findings indicates that there is a link between TMEM97 and chronic inflammation in obesity. en-copyright= kn-copyright= en-aut-name=TentaMasafumi en-aut-sei=Tenta en-aut-mei=Masafumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=EguchiJun en-aut-sei=Eguchi en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WadaJun en-aut-sei=Wada en-aut-mei=Jun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Department of Nephrology, Rheumatology, Endocrinology, and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Nephrology, Rheumatology, Endocrinology, and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Nephrology, Rheumatology, Endocrinology, and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=adipose tissue kn-keyword=adipose tissue en-keyword=skeletal muscle kn-keyword=skeletal muscle en-keyword=obesity kn-keyword=obesity END start-ver=1.4 cd-journal=joma no-vol=70 cd-vols= no-issue=2 article-no= start-page=146 end-page=154 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Development of Scaled-Up Synthetic Method for Retinoid X Receptor Agonist NEt-3IB Contributing to Sustainable Development Goals en-subtitle= kn-subtitle= en-abstract= kn-abstract=Abstract Small-molecular drugs, which are generally inexpensive compared with biopharmaceuticals and can often be taken orally, may contribute to the Sustainable Development Goals (SDGs) adopted by the United Nations. We previously reported the retinoid X receptor (RXR) agonist 4-(ethyl(3-isobutoxy-4-isopropylphenyl)amino)benzoic acid (NEt-3IB, 1) as a small-molecular drug candidate to replace biopharmaceuticals for the treatment of inflammatory bowel disease. The previous synthetic method to 1 required a large amount of organic solvent and extensive purification. In line with the SDGs, we aimed to develop an environmentally friendly, inexpensive method for the large-scale synthesis of 1. The developed method requires only a hydrophobic ether and EtOH as reaction and extraction solvents. The product was purified by recrystallization twice to afford 99% pure 1 at 100?mmol scale in about 30% yield. The optimized process showed a 35-fold improvement of the E-factor (an index of environmental impact) compared to the original method. This work, which changes the solvent used to environmentally preferable ones based on the existing synthetic method for 1, illustrates how synthetic methods for small-molecular drugs can be adapted and improved to contribute to the SDGs. en-copyright= kn-copyright= en-aut-name=TakamuraYuta en-aut-sei=Takamura en-aut-mei=Yuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MorishitaKen-ichi en-aut-sei=Morishita en-aut-mei=Ken-ichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KikuzawaShota en-aut-sei=Kikuzawa en-aut-mei=Shota kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=WatanabeMasaki en-aut-sei=Watanabe en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KakutaHiroki en-aut-sei=Kakuta en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=167 cd-vols= no-issue= article-no= start-page=923 end-page=929 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220203 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A novel victorivirus from the phytopathogenic fungus Neofusicoccum parvum en-subtitle= kn-subtitle= en-abstract= kn-abstract=Neofusicoccum parvum is an important plant-pathogenic ascomycetous fungus that causes trunk diseases in a variety of plants. A limited number of reports on mycoviruses from this fungus are available. Here, we report the characterization of a novel victorivirus, Neofusicoccum parvum victorivirus 3 (NpVV3). An agarose gel dsRNA profile of a Pakistani strain of N. parvum, NFN, showed a band of similar to 5 kbp that was not detectable in Japanese strains of N. parvum. Taking a high-throughput and Sanger sequencing approach, the complete genome sequence of NpVV3 was determined to be 5226 bp in length with two open reading frames (ORF1 and ORF2) that encode a capsid protein (CP) and an RNA-dependent RNA polymerase (RdRP). The RdRP appears to be translated by a stop/restart mechanism facilitated by the junction sequence AUGucUGA, as is found in some other victoriviruses. BLASTp searches showed that NpVV3 CP and RdRP share the highest amino acid sequence identity (80.5% and 72.4%, respectively) with the corresponding proteins of NpVV1 isolated from a French strain of N. parvum. However, NpVV3 was found to be different from NpVV1 in its terminal sequences and the stop/restart facilitator sequence. NpVV3 particles similar to 35 nm in diameter were partially purified and used to infect an antiviral-RNA-silencing-deficient strain (Delta cl2) of an experimental ascomycetous fungal host, Cryphonectria parasitica. NpVV3 showed symptomless infection in the new host strain. en-copyright= kn-copyright= en-aut-name=KhanHaris Ahmed en-aut-sei=Khan en-aut-mei=Haris Ahmed kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SatoYukiyo en-aut-sei=Sato en-aut-mei=Yukiyo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KondoHideki en-aut-sei=Kondo en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=JamalAtif en-aut-sei=Jamal en-aut-mei=Atif kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=BhattiMuhammad Faraz en-aut-sei=Bhatti en-aut-mei=Muhammad Faraz kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SuzukiNobuhiro en-aut-sei=Suzuki en-aut-mei=Nobuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST) kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=4 en-affil=Crop Diseases Research Institute, National Agricultural Research Centre kn-affil= affil-num=5 en-affil=Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST) kn-affil= affil-num=6 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=111 cd-vols= no-issue= article-no= start-page=7 end-page=14 dt-received= dt-revised= dt-accepted= dt-pub-year=2022 dt-pub=20220201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on antitumor enzyme l-lysine ƒΏ-oxidase from Trichoderma viride kn-title=Ž…σ‹ΫTrichoderma viride—R—ˆ‚̍RŽξᇐ«y‘fL-ƒŠƒVƒ“ ƒΏ-ƒIƒLƒVƒ_[ƒ[‚ΙŠΦ‚·‚ιŒ€‹† en-subtitle= kn-subtitle= en-abstract= kn-abstract=L-Lysine ƒΏ-oxidase (LysOX) from Trichoderma viride is a homodimeric flavoenzyme that catalyzes the oxidative deamination of L-Lysine to produce ƒΏ-keto-ƒΓ-aminocaproate with ammonia and hydrogen per-oxide. LysOX inhibited the growth of cancer cells but showed relatively low toxicity for normal cells. The full-length cDNA consists of 2,119 bp, and encodes a long N-terminal propeptide composed of 77 resi-dues (Met1-Arg77) and the mature protein (Ala78-Ile617). The LysOX gene was heterologously expressed in Streptomyces lividans TK24 or Escherichia coli SoluBL21. The enzymatic properties of the purified recombinant LysOX, such as substrate specificity, kinetic parameters and thermal stability, are the same as those of the native LysOX. The LysOX precursor (prLysOX) expressed in E. coli shows weak enzymatic activity and is activated by proteolytic processing. The crystal structure of prLysOX revealed that the propeptide of prLysOX indirectly changes the active site structure to inhibit enzyme activity. Moreover, the crystal structures of LysOX and its L-Lysine complex revealed that the hydrogen bonding network formed by Asp212, Asp315 and Ala440 with two water molecules is responsible for the recogni-tion of the ƒΓ-amino group of L-Lysine. In addition, a narrow substrate-binding site and acidic surface at the active site entrance both contribute to strict substrate specificity. Mutational analysis demonstrated that Asp212 and Asp315 are essential for substrate recognition, and the D212A/D315A LysOX prefers aromatic amino acids. Furthermore, the structural basis of the substrate specificity change has also been revealed by the structural analysis of the D212A/D315A LysOX and its substrate complexes. en-copyright= kn-copyright= en-aut-name=SaitoMasaya en-aut-sei=Saito en-aut-mei=Masaya kn-aut-name=βV“‘‰λΖ kn-aut-sei=βV“‘ kn-aut-mei=‰λΖ aut-affil-num=1 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name=ˆξŠ_Œ«“ρ kn-aut-sei=ˆξŠ_ kn-aut-mei=Œ«“ρ aut-affil-num=2 ORCID= affil-num=1 en-affil=Course of Agrochemical Bioscience kn-affil=‰ͺŽR‘εŠw”_Šw•””_Œ|‰»ŠwƒR[ƒX affil-num=2 en-affil= kn-affil=ŠwpŒ€‹†‰@ŠΒ‹«Ά–½‰ΘŠwŠwˆζ en-keyword=L-lysine ƒΏ-oxidase kn-keyword=L-lysine ƒΏ-oxidase en-keyword=antitumor enzyme kn-keyword=antitumor enzyme en-keyword=substrate recognition kn-keyword=substrate recognition en-keyword=X-ray crystallography kn-keyword=X-ray crystallography en-keyword=enzyme activity regulation kn-keyword=enzyme activity regulation END start-ver=1.4 cd-journal=joma no-vol=75 cd-vols= no-issue=6 article-no= start-page=671 end-page=675 dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=202112 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Multiple Roles of Histidine-Rich Glycoprotein in Vascular Homeostasis and Angiogenesis en-subtitle= kn-subtitle= en-abstract= kn-abstract=Histidine-rich glycoprotein (HRG) is a 75 kDa plasma protein that is synthesized in the liver of many verte-brates and present in their plasma at relatively high concentrations of 100-150 ƒΚg/mL. HRG is an abundant and well-characterized protein having a multidomain structure that enable it to interact with many ligands, func-tion as an adaptor molecule, and participate in numerous physiological and pathological processes. As a plasma protein, HRG has been reported to regulate vascular biology, including coagulation, fibrinolysis and angiogenesis, through its binding with several ligands (heparin, FXII, fibrinogen, thrombospondin, and plas-minogen) and interaction with many types of cells (endothelial cells, erythrocytes, neutrophils and platelets). This review aims to summarize the roles of HRG in maintaining vascular homeostasis and regulating angiogen-esis in various pathological conditions. en-copyright= kn-copyright= en-aut-name=GaoShangze en-aut-sei=Gao en-aut-mei=Shangze kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NishiboriMasahiro en-aut-sei=Nishibori en-aut-mei=Masahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= affil-num=1 en-affil=Department of Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=histidine-rich glycoprotein kn-keyword=histidine-rich glycoprotein en-keyword=vascular biology kn-keyword=vascular biology en-keyword=coagulation kn-keyword=coagulation en-keyword=angiogenesis kn-keyword=angiogenesis END start-ver=1.4 cd-journal=joma no-vol=12 cd-vols= no-issue= article-no= start-page=647684 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20210810 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Purification, Characterization, and Gene Expression of Rice Endo-beta-N-Acetylglucosaminidase, Endo-Os en-subtitle= kn-subtitle= en-abstract= kn-abstract=In the endoplasmic reticulum-associated degradation system of plant and animal cells, high-mannose type free N-glycans (HMT-FNGs) are produced from misfolded glycoproteins prior to proteasomal degradation, and two enzymes, cytosolic peptide:N-glycanase (cPNGase) and endo-beta-N-acetylglucosaminidase (endo-beta-GlcNAc-ase), are involved in the deglycosylation. Although the physiological functions of these FNGs in plant growth and development remain to be elucidated, detailed characterization of cPNGase and endo-beta-GlcNAc-ase is required. In our previous work, we described the purification, characterization, and subcellular distribution of some plant endo-beta-GlcNAc-ases and preliminarily reported the gene information of rice endo-beta-GlcNAc-ase (Endo-Os). Furthermore, we analyzed the changes in gene expression of endo-beta-GlcNAc-ase during tomato fruit maturation and constructed a mutant line of Arabidopsis thaliana, in which the two endo-beta-GlcNAc-ase genes were knocked-out based on the Endo-Os gene. In this report, we describe the purification, characterization, amino acid sequence, and gene cloning of Endo-Os in detail. Purified Endo-Os, with an optimal pH of 6.5, showed high activity for high-mannose type N-glycans bearing the Man alpha 1-2Man alpha 1-3Man beta 1 unit; this substrate specificity was almost the same as that of other plant endo-beta-GlcNAc-ases, suggesting that Endo-Os plays a critical role in the production of HTM-FNGs in the cytosol. Electrospray ionization-mass spectrometry analysis of the tryptic peptides revealed 17 internal amino acid sequences, including the C terminus; the N-terminal sequence could not be identified due to chemical modification. These internal amino acid sequences were consistent with the amino acid sequence (UniProt ID: Q5W6R1) deduced from the Oryza sativa cDNA clone AK112067 (gene ID: Os05g0346500). Recombinant Endo-Os expressed in Escherichia coli using cDNA showed the same enzymatic properties as those of native Endo-Os. en-copyright= kn-copyright= en-aut-name=MaedaMegumi en-aut-sei=Maeda en-aut-mei=Megumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OkamotoNaoko en-aut-sei=Okamoto en-aut-mei=Naoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=ArakiNode en-aut-sei=Araki en-aut-mei=Node kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KimuraYoshinobu en-aut-sei=Kimura en-aut-mei=Yoshinobu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=3 en-affil=Department of Tumor Genetics and Biology, Graduate School of Medical Sciences, Kumamoto University kn-affil= affil-num=4 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword=endo-beta-N-acetylglucosaminidase kn-keyword=endo-beta-N-acetylglucosaminidase en-keyword=free N-glycans kn-keyword=free N-glycans en-keyword=Oryza sativa kn-keyword=Oryza sativa en-keyword=ER associated degradation kn-keyword=ER associated degradation en-keyword=peptide:N-glycanase kn-keyword=peptide:N-glycanase END start-ver=1.4 cd-journal=joma no-vol=169 cd-vols= no-issue=5 article-no= start-page=585 end-page=599 dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=202112 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A sweet protein monellin as a non-antibody scaffold for synthetic binding proteins en-subtitle= kn-subtitle= en-abstract= kn-abstract=Synthetic binding proteins that have the ability to bind with molecules can be generated using various protein domains as non-antibody scaffolds. These designer proteins have been used widely in research studies, as their properties overcome the disadvantages of using antibodies. Here, we describe the first application of a phage display to generate synthetic binding proteins using a sweet protein, monellin, as a non-antibody scaffold. Single-chain monellin (scMonellin), in which two polypeptide chains of natural monellin are connected by a short linker, has two loops on one side of the molecule. We constructed phage display libraries of scMonellin, in which the amino acid sequence of the two loops is diversified. To validate the performance of these libraries, we sorted them against the folding mutant of the green fluorescent protein variant (GFPuv) and yeast small ubiquitin-related modifier. We successfully obtained scMonellin variants exhibiting moderate but significant affinities for these target proteins. Crystal structures of one of the GFPuv-binding variants in complex with GFPuv revealed that the two diversified loops were involved in target recognition. scMonellin, therefore, represents a promising non-antibody scaffold in the design and generation of synthetic binding proteins. We termed the scMonellin-derived synthetic binding proteins eSWEEPinsf. en-copyright= kn-copyright= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NakamuraKazuaki en-aut-sei=Nakamura en-aut-mei=Kazuaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 1-1-1, Tsushima-naka, Kita-ku, Okayama 700-8530, Japan kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 1-1-1, Tsushima-naka, Kita-ku, Okayama 700-8530, Japan kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 1-1-1, Tsushima-naka, Kita-ku, Okayama 700-8530, Japan kn-affil= en-keyword=combinatorial library kn-keyword=combinatorial library en-keyword=non-antibody scaffold kn-keyword=non-antibody scaffold en-keyword=phage display kn-keyword=phage display en-keyword=single-chain monellin kn-keyword=single-chain monellin en-keyword=synthetic binding proteins kn-keyword=synthetic binding proteins END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=1 article-no= start-page=266 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20210710 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Genome sequence analysis of new plum pox virus isolates from Japan en-subtitle= kn-subtitle= en-abstract= kn-abstract=Objective To find mutations that may have recently occurred in Plum pox virus (PPV), we collected six PPV-infected plum/peach trees from the western part of Japan and one from the eastern part. After sequencing the full-length PPV genomic RNAs, we compared the amino acid sequences with representative isolates of each PPV strain. Results All new isolates were found to belong to the PPV-D strain: the six isolates collected from western Japan were identified as the West-Japan strain while the one collected from eastern Japan as the East-Japan strain. Amino acid sequence analysis of these seven isolates suggested that the 1407th and 1529th amino acid residues are characteristic of the West-Japan and the East-Japan strains, respectively. Comparing them with the corresponding amino acid residues of the 47 non-Japanese PPV-D isolates revealed that these amino acid residues are undoubtedly unique. A further examination of the relevant amino acid residues of the other 210 PPV-D isolates collected in Japan generated a new hypothesis regarding the invasion route from overseas and the subsequent diffusion route within Japan: a PPV-D strain might have invaded the western part of Japan from overseas and spread throughout Japan. en-copyright= kn-copyright= en-aut-name=MoriTomoaki en-aut-sei=Mori en-aut-mei=Tomoaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=WarnerChiaki en-aut-sei=Warner en-aut-mei=Chiaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OhnoSerika en-aut-sei=Ohno en-aut-mei=Serika kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MoriKoichi en-aut-sei=Mori en-aut-mei=Koichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TobimatsuTakamasa en-aut-sei=Tobimatsu en-aut-mei=Takamasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SeraTakashi en-aut-sei=Sera en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=4 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=5 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=6 en-affil=Department of Applied Chemistry and Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= en-keyword=Plum pox virus kn-keyword=Plum pox virus en-keyword=Complete genome sequence kn-keyword=Complete genome sequence en-keyword=Phylogenetic analysis kn-keyword=Phylogenetic analysis en-keyword=Sequence alignment analysis kn-keyword=Sequence alignment analysis en-keyword=Genetic variation kn-keyword=Genetic variation END start-ver=1.4 cd-journal=joma no-vol=22 cd-vols= no-issue=13 article-no= start-page=7235 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20210705 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Citric Acid-Mediated Abiotic Stress Tolerance in Plants en-subtitle= kn-subtitle= en-abstract= kn-abstract=Several recent studies have shown that citric acid/citrate (CA) can confer abiotic stress tolerance to plants. Exogenous CA application leads to improved growth and yield in crop plants under various abiotic stress conditions. Improved physiological outcomes are associated with higher photosynthetic rates, reduced reactive oxygen species, and better osmoregulation. Application of CA also induces antioxidant defense systems, promotes increased chlorophyll content, and affects secondary metabolism to limit plant growth restrictions under stress. In particular, CA has a major impact on relieving heavy metal stress by promoting precipitation, chelation, and sequestration of metal ions. This review summarizes the mechanisms that mediate CA-regulated changes in plants, primarily CA's involvement in the control of physiological and molecular processes in plants under abiotic stress conditions. We also review genetic engineering strategies for CA-mediated abiotic stress tolerance. Finally, we propose a model to explain how CA's position in complex metabolic networks involving the biosynthesis of phytohormones, amino acids, signaling molecules, and other secondary metabolites could explain some of its abiotic stress-ameliorating properties. This review summarizes our current understanding of CA-mediated abiotic stress tolerance and highlights areas where additional research is needed. en-copyright= kn-copyright= en-aut-name=Tahjib-Ul-ArifMd. en-aut-sei=Tahjib-Ul-Arif en-aut-mei=Md. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ZahanMst, Ishrat en-aut-sei=Zahan en-aut-mei=Mst, Ishrat kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KarimMd. Masudul en-aut-sei=Karim en-aut-mei=Md. Masudul kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ImranShahin en-aut-sei=Imran en-aut-mei=Shahin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=HunterCharles T. en-aut-sei=Hunter en-aut-mei=Charles T. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=IslamMd. Saiful en-aut-sei=Islam en-aut-mei=Md. Saiful kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MiaMd. Ashik en-aut-sei=Mia en-aut-mei=Md. Ashik kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=HannanMd. Abdul en-aut-sei=Hannan en-aut-mei=Md. Abdul kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=RhamanMohammad Saidur en-aut-sei=Rhaman en-aut-mei=Mohammad Saidur kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=HossainMd. Afzal en-aut-sei=Hossain en-aut-mei=Md. Afzal kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=BresticMarian en-aut-sei=Brestic en-aut-mei=Marian kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=SkalickyMilan en-aut-sei=Skalicky en-aut-mei=Milan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=MurataYoshiyuki en-aut-sei=Murata en-aut-mei=Yoshiyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=Plant Breeding Division, Bangladesh Rice Research Institute kn-affil= affil-num=3 en-affil=Department of Crop Botany, Bangladesh Agricultural University kn-affil= affil-num=4 en-affil=Department of Agronomy, Khulna Agricultural University kn-affil= affil-num=5 en-affil=Chemistry Research Unit, United States Department of Agriculture?Agricultural Research Service kn-affil= affil-num=6 en-affil=Department of Fisheries, Bangamata Sheikh Fojilatunnesa Mujib Science and Technology University kn-affil= affil-num=7 en-affil=Department of Crop Botany, Bangladesh Agricultural University kn-affil= affil-num=8 en-affil=Department of Biochemistry and Molecular Biology, Bangladesh Agricultural University kn-affil= affil-num=9 en-affil=Department of Seed Science and Technology, Bangladesh Agricultural University kn-affil= affil-num=10 en-affil=Department of Biochemistry and Molecular Biology, Bangladesh Agricultural University kn-affil= affil-num=11 en-affil=Department of Plant Physiology, Slovak University of Agriculture kn-affil= affil-num=12 en-affil=Department of Botany and Plant Physiology, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague kn-affil= affil-num=13 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword=citrate kn-keyword=citrate en-keyword=heavy metal stress kn-keyword=heavy metal stress en-keyword=drought stress kn-keyword=drought stress en-keyword=antioxidant kn-keyword=antioxidant en-keyword=reactive oxygen species kn-keyword=reactive oxygen species en-keyword=salinity kn-keyword=salinity en-keyword=aluminum toxicity kn-keyword=aluminum toxicity END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=6 article-no= start-page=1442 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20210316 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Antibacterial Effect of Amino Acid-Silver Complex Loaded Montmorillonite Incorporated in Dental Acrylic Resin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Several dental materials contain silver for antibacterial effect, however the effect is relatively low. The reason for the lower antibacterial efficacy of silver is considered to be the fact that silver ions bind to chloride ions in saliva. To develop new effective silver antibacterial agents that can be useful in the mouth, we synthesized two novel amino acid (methionine or histidine)-silver complexes (Met or His-Ag) loaded with montmorillonite (Mont) and analyzed their antibacterial efficacy. At first the complexes were characterized using nuclear magnetic resonance (NMR), and amino acid-Ag complex-loaded Mont (amino acid-Ag-Mont) were characterized using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The antibacterial efficacy of these materials in dental acrylic resin was then investigated by bacterial growth measurement using a spectrophotometer. As controls, commercially available silver-loaded zeolite and silver-zirconium phosphate were also tested. Dental acrylic resin incorporating His-Ag-Mont strongly inhibited Streptococcus mutans growth. This was explained by the fact that His-Ag complex revealed the highest amounts of silver ions in the presence of chloride. The structure of the amino acid-Ag complexes affected the silver ion presence in chloride and the antibacterial efficacy. His-Ag-Mont might be used as antibacterial agents for dental materials. en-copyright= kn-copyright= en-aut-name=YoshiharaKumiko en-aut-sei=Yoshihara en-aut-mei=Kumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NagaokaNoriyuki en-aut-sei=Nagaoka en-aut-mei=Noriyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=UmenoAya en-aut-sei=Umeno en-aut-mei=Aya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SonodaAkinari en-aut-sei=Sonoda en-aut-mei=Akinari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ObikaHideki en-aut-sei=Obika en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=YoshidaYasuhiro en-aut-sei=Yoshida en-aut-mei=Yasuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=Van MeerbeekBart en-aut-sei=Van Meerbeek en-aut-mei=Bart kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MakitaYoji en-aut-sei=Makita en-aut-mei=Yoji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Department of Pathology & Experimental Medicine, Dentistry and Pharmaceutical Sciences, Graduate School of Medicine, Okayama University Hospital kn-affil= affil-num=2 en-affil=Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School kn-affil= affil-num=3 en-affil=Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) kn-affil= affil-num=4 en-affil=Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) kn-affil= affil-num=5 en-affil=Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) kn-affil= affil-num=6 en-affil=Department of Biomaterials and Bioengineering, Faculty of Dental Medicine, Hokkaido University kn-affil= affil-num=7 en-affil=KU Leuven (University of Leuven) Department of Oral Health Research, BIOMAT & University Hospitals Leuven kn-affil= affil-num=8 en-affil=Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) kn-affil= en-keyword=montmorillonite kn-keyword=montmorillonite en-keyword=amino acid kn-keyword=amino acid en-keyword=antibacterial kn-keyword=antibacterial en-keyword=Streptococcus mutans kn-keyword=Streptococcus mutans en-keyword=nuclear magnetic resonance kn-keyword=nuclear magnetic resonance en-keyword=X-ray diffraction kn-keyword=X-ray diffraction END start-ver=1.4 cd-journal=joma no-vol=5 cd-vols= no-issue= article-no= start-page=100044 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20211231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Structural basis of enzyme activity regulation by the propeptide of l-lysine ƒΏ-oxidase precursor from Trichoderma viride en-subtitle= kn-subtitle= en-abstract= kn-abstract=Harmuful proteins are usually synthesized as inactive precursors and are activated by proteolytic processing. l-Amino acid oxidase (LAAO) is a flavoenzyme that catalyzes the oxidative deamination of l-amino acid to produce a 2-oxo acid with ammonia and highly toxic hydrogen peroxide and, therefore, is expressed as a precursor. The LAAO precursor shows significant variation in size and the cleavage pattern for activation. However, the molecular mechanism of how the propeptide suppresses the enzyme activity remains unclear except for deaminating/decarboxylating Pseudomonasl-phenylalanine oxidase (PAO), which has a short N-terminal propeptide composed of 14 residues. Here we show the inactivation mechanism of the l-lysine oxidase (LysOX) precursor (prLysOX), which has a long N-terminal propeptide composed of 77 residues, based on the crystal structure at 1.97?? resolution. The propeptide of prLysOX indirectly changes the active site structure to inhibit the enzyme activity. prLysOX retains weak enzymatic activity with strict specificity for l-lysine and shows raised activity in acidic conditions. The structures of prLysOX crystals that soaked in a solution with various concentrations of l-lysine have revealed that prLysOX can adopt two conformations; one is the inhibitory form, and the other is very similar to mature LysOX. The propeptide region of the latter form is disordered, and l-lysine is bound to the latter form. These results indicate that prLysOX uses a different strategy from PAO to suppress the enzyme activity and suggest that prLysOX can be activated quickly in response to the environmental change without proteolytic processing. en-copyright= kn-copyright= en-aut-name=KitagawaMasaki en-aut-sei=Kitagawa en-aut-mei=Masaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ItoNanako en-aut-sei=Ito en-aut-mei=Nanako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MatsumotoYuya en-aut-sei=Matsumoto en-aut-mei=Yuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SaitoMasaya en-aut-sei=Saito en-aut-mei=Masaya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TamuraTakashi en-aut-sei=Tamura en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KusakabeHitoshi en-aut-sei=Kusakabe en-aut-mei=Hitoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=ImadaKatsumi en-aut-sei=Imada en-aut-mei=Katsumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Department of Macromolecular Science, Graduate School of Science, Osaka University kn-affil= affil-num=2 en-affil=Department of Macromolecular Science, Graduate School of Science, Osaka University kn-affil= affil-num=3 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=4 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=5 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=6 en-affil=Enzyme Sensor Co., Ltd. kn-affil= affil-num=7 en-affil=Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=8 en-affil=Department of Macromolecular Science, Graduate School of Science, Osaka University kn-affil= en-keyword=L-Lysine ƒΏ-oxidase kn-keyword=L-Lysine ƒΏ-oxidase en-keyword=Crystal structure kn-keyword=Crystal structure en-keyword=Precursor kn-keyword=Precursor en-keyword=Substrate recognition kn-keyword=Substrate recognition END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=1 article-no= start-page=208 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2021 dt-pub=20210104 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Tryptophan and Kynurenine Enhances the Stemness and Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro and In Vivo en-subtitle= kn-subtitle= en-abstract= kn-abstract=Aging tissues present a progressive decline in homeostasis and regenerative capacities, which has been associated with degenerative changes in tissue-specific stem cells and stem cell niches. We hypothesized that amino acids could regulate the stem cell phenotype and differentiation ability of human bone marrow-derived mesenchymal stromal cells (hBMSCs). Thus, we performed a screening of 22 standard amino acids and found that D-tryptophan (10 mu M) increased the number of cells positive for the early stem cell marker SSEA-4, and the gene expression levels of OCT-4, NANOG, and SOX-2 in hBMSCs. Comparison between D- and L-tryptophan isomers showed that the latter presents a stronger effect in inducing the mRNA levels of Oct-4 and Nanog, and in increasing the osteogenic differentiation of hBMSCs. On the other hand, L-tryptophan suppressed adipogenesis. The migration and colony-forming ability of hBMSCs were also enhanced by L-tryptophan treatment. In vivo experiments delivering L-tryptophan (50 mg/kg/day) by intraperitoneal injections for three weeks confirmed that L-tryptophan significantly increased the percentage of cells positive for SSEA-4, mRNA levels of Nanog and Oct-4, and the migration and colony-forming ability of mouse BMSCs. L-kynurenine, a major metabolite of L-tryptophan, also induced similar effects of L-tryptophan in enhancing stemness and osteogenic differentiation of BMSCs in vitro and in vivo, possibly indicating the involvement of the kynurenine pathway as the downstream signaling of L-tryptophan. Finally, since BMSCs migrate to the wound healing site to promote bone healing, surgical defects of 1 mm in diameter were created in mouse femur to evaluate bone formation after two weeks of L-tryptophan or L-kynurenine injection. Both L-tryptophan and L-kynurenine accelerated bone healing compared to the PBS-injected control group. In summary, L-tryptophan enhanced the stemness and osteoblastic differentiation of BMSCs and may be used as an essential factor to maintain the stem cell properties and accelerate bone healing and/or prevent bone loss. en-copyright= kn-copyright= en-aut-name=PhamHai Thanh en-aut-sei=Pham en-aut-mei=Hai Thanh kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OnoMitsuaki en-aut-sei=Ono en-aut-mei=Mitsuaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HaraEmilio Satoshi en-aut-sei=Hara en-aut-mei=Emilio Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=NguyenHa Thi Thu en-aut-sei=Nguyen en-aut-mei=Ha Thi Thu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=DangAnh Tuan en-aut-sei=Dang en-aut-mei=Anh Tuan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=DoHang Thuy en-aut-sei=Do en-aut-mei=Hang Thuy kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KomoriTaishi en-aut-sei=Komori en-aut-mei=Taishi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TosaIkue en-aut-sei=Tosa en-aut-mei=Ikue kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=Hazehara-KunitomoYuri en-aut-sei=Hazehara-Kunitomo en-aut-mei=Yuri kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=YoshiokaYuya en-aut-sei=Yoshioka en-aut-mei=Yuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=OidaYasutaka en-aut-sei=Oida en-aut-mei=Yasutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=AkiyamaKentaro en-aut-sei=Akiyama en-aut-mei=Kentaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=KubokiTakuo en-aut-sei=Kuboki en-aut-mei=Takuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= affil-num=1 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=9 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=10 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=11 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=12 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=13 en-affil=Department of Oral Rehabilitation and Regenerative Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=amino acid kn-keyword=amino acid en-keyword=mesenchymal stromal cells kn-keyword=mesenchymal stromal cells en-keyword=stemness kn-keyword=stemness en-keyword=tryptophan kn-keyword=tryptophan en-keyword=kynurenine kn-keyword=kynurenine en-keyword=osteogenesis kn-keyword=osteogenesis en-keyword=adipogenesis kn-keyword=adipogenesis en-keyword=screening kn-keyword=screening en-keyword=injury/fracture healing kn-keyword=injury/fracture healing en-keyword=anabolics kn-keyword=anabolics END start-ver=1.4 cd-journal=joma no-vol=35 cd-vols= no-issue=4 article-no= start-page=ME20114 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=2020 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Requirement of ƒΑ-Aminobutyric Acid Chemotaxis for Virulence of Pseudomonas syringae pv. tabaci 6605 en-subtitle= kn-subtitle= en-abstract= kn-abstract=ƒΑ-Aminobutyric acid (GABA) is a widely distributed non-proteinogenic amino acid that accumulates in plants under biotic and abiotic stress conditions. Recent studies suggested that GABA also functions as an intracellular signaling molecule in plants and in signals mediating interactions between plants and phytopathogenic bacteria. However, the molecular mechanisms underlying GABA responses to bacterial pathogens remain unknown. In the present study, a GABA receptor, named McpG, was conserved in the highly motile plant-pathogenic bacteria Pseudomonas syringae pv. tabaci 6605 (Pta6605). We generated a deletion mutant of McpG to further investigate its involvement in GABA chemotaxis using quantitative capillary and qualitative plate assays. The wild-type strain of Pta6605 was attracted to GABA, while the ƒ’mcpG mutant abolished chemotaxis to 10? ?mM GABA. However, ƒ’mcpG retained chemotaxis to proteinogenic amino acids and succinic semialdehyde, a structural analog of GABA. Furthermore, ƒ’mcpG was unable to effectively induce disease on host tobacco plants in three plant inoculation assays: flood, dip, and infiltration inoculations. These results revealed that the GABA sensing of Pta6605 is important for the interaction of Pta6605 with its host tobacco plant. en-copyright= kn-copyright= en-aut-name=TumewuStephany Angelia en-aut-sei=Tumewu en-aut-mei=Stephany Angelia kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsuiHidenori en-aut-sei=Matsui en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamamotoMikihiro en-aut-sei=Yamamoto en-aut-mei=Mikihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=NoutoshiYoshiteru en-aut-sei=Noutoshi en-aut-mei=Yoshiteru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ToyodaKazuhiro en-aut-sei=Toyoda en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=6 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword= bacterial virulence kn-keyword= bacterial virulence en-keyword=chemotaxis kn-keyword=chemotaxis en-keyword=GABA kn-keyword=GABA en-keyword=plant-microbe interaction kn-keyword=plant-microbe interaction en-keyword=Pseudomonas kn-keyword=Pseudomonas END start-ver=1.4 cd-journal=joma no-vol=9 cd-vols= no-issue=10 article-no= start-page=2149 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200923 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=In Vitro Studies to Define the Cell-Surface and Intracellular Targets of Polyarginine-Conjugated Sodium Borocaptate as a Potential Delivery Agent for Boron Neutron Capture Therapy en-subtitle= kn-subtitle= en-abstract= kn-abstract=Boron neutron capture therapy (BNCT) requires pharmaceutical innovations and molecular-based evidence of effectiveness to become a standard cancer therapeutic in the future. Recently, in Japan, 4-borono-L-phenylalanine (BPA) was approved as a boron agent for BNCT against head and neck (H&N) cancers. H&N cancer appears to be a suitable target for BPA-BNCT, because the expression levels of L-type amino acid transporter 1 (LAT1), one of the amino acid transporters responsible for BPA uptake, are elevated in most cases of H&N cancer. However, in other types of cancer including malignant brain tumors, LAT1 is not always highly expressed. To expand the possibility of BNCT for these cases, we previously developed poly-arginine peptide (polyR)-conjugated mercaptoundecahydrododecaborate (BSH). PolyR confers the cell membrane permeability and tumor selectivity of BSH. However, the molecular determinants for the properties are not fully understood. In this present study, we have identified the cluster of differentiation 44 (CD44) protein and translational machinery proteins as a major cell surface target and intracellular targets of BSH-polyR, respectively. CD44, also known as a stem cell-associated maker in various types of cancer, is required for the cellular uptake of polyR-conjugated molecules. We showed that BSH-polyR was predominantly delivered to a CD44(High) cell population of cancer cells. Once delivered, BSH-polyR interacted with the translational machinery components, including the initiation factors, termination factors, and poly(A)-biding protein (PABP). As a proof of principle, we performed BSH-polyR-based BNCT against glioma stem-like cells and revealed that BSH-polyR successfully induced BNCT-dependent cell death specifically in CD44(High) cells. Bioinformatics analysis indicated that BSH-polyR would be suitable for certain types of malignant tumors. Our results shed light on the biochemical properties of BSH-polyR, which may further contribute to the therapeutic optimization of BSH-BNCT in the future. en-copyright= kn-copyright= en-aut-name=FujimuraAtsushi en-aut-sei=Fujimura en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YasuiSeiji en-aut-sei=Yasui en-aut-mei=Seiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IgawaKazuyo en-aut-sei=Igawa en-aut-mei=Kazuyo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=UedaAi en-aut-sei=Ueda en-aut-mei=Ai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=WatanabeKaori en-aut-sei=Watanabe en-aut-mei=Kaori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HanafusaTadashi en-aut-sei=Hanafusa en-aut-mei=Tadashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=IchikawaYasuaki en-aut-sei=Ichikawa en-aut-mei=Yasuaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=YoshihashiSachiko en-aut-sei=Yoshihashi en-aut-mei=Sachiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=TsuchidaKazuki en-aut-sei=Tsuchida en-aut-mei=Kazuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=KamiyaAtsunori en-aut-sei=Kamiya en-aut-mei=Atsunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=FuruyaShuichi en-aut-sei=Furuya en-aut-mei=Shuichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=3 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=4 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=5 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=6 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=7 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= affil-num=8 en-affil=Graduate School of Engineering, Nagoya University kn-affil= affil-num=9 en-affil=Graduate School of Engineering, Nagoya University kn-affil= affil-num=10 en-affil=Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=11 en-affil=Neutron Therapy Research Center, Okayama University kn-affil= en-keyword=boron neutron capture therapy (BNCT) kn-keyword=boron neutron capture therapy (BNCT) en-keyword=BSH-polyR kn-keyword=BSH-polyR en-keyword=CD44 kn-keyword=CD44 en-keyword=translational machinery kn-keyword=translational machinery en-keyword=bioinformatics kn-keyword=bioinformatics END start-ver=1.4 cd-journal=joma no-vol=117 cd-vols= no-issue=10 article-no= start-page=101103 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200909 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Super-chiral vibrational spectroscopy with metasurfaces for high-sensitive identification of alanine enantiomers en-subtitle= kn-subtitle= en-abstract= kn-abstract=Chiral nature of an enantiomer can be characterized by circular dichroism (CD) spectroscopy, but such a technique usually suffers from weak signal even with a sophisticated optical instrument. Recent demonstrations of plasmonic metasurfaces showed that chiroptical interaction of molecules can be engineered, thereby greatly simplifying a measurement system with high sensing capability. Here, by exploiting super-chiral field in a metasurface, we experimentally demonstrate high-sensitive vibrational CD spectroscopy of alanine enantiomers, the smallest chiral amino acid. Under linearly polarized excitation, the metasurface consisting of an array of staggered Au nano-rods selectively produces the left- and right-handed super-chiral fields at 1600?cm?1, which spectrally overlaps with the functional group vibrations of alanine. In the Fourier-transform infrared spectrometer measurements, the mirror symmetric CD spectra of D- and L-alanine are clearly observed depending on the handedness of the metasurface, realizing the reliable identification of small chiral molecules. The corresponding numerical simulations reveal the underlying resonant chiroptical interaction of plasmonic modes of the metasurface and vibrational modes of alanine. Our approach demonstrates a high-sensitive vibrational CD spectroscopic technique, opening up a reliable chiral sensing platform for advanced infrared inspection technologies. en-copyright= kn-copyright= en-aut-name=IidaTakumi en-aut-sei=Iida en-aut-mei=Takumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IshikawaAtsushi en-aut-sei=Ishikawa en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TanakaTakuo en-aut-sei=Tanaka en-aut-mei=Takuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MuranakaAtsuya en-aut-sei=Muranaka en-aut-mei=Atsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=UchiyamaMasanobu en-aut-sei=Uchiyama en-aut-mei=Masanobu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HayashiYasuhiko en-aut-sei=Hayashi en-aut-mei=Yasuhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=TsurutaKenji en-aut-sei=Tsuruta en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Department of Electrical and Electronic Engineering, Okayama University kn-affil= affil-num=2 en-affil=Department of Electrical and Electronic Engineering, Okayama University kn-affil= affil-num=3 en-affil=Metamaterials Laboratory, RIKEN Cluster for Pioneering Research kn-affil= affil-num=4 en-affil=Advanced Elements Chemistry Laboratory, RIKEN Cluster for Pioneering Research kn-affil= affil-num=5 en-affil=Advanced Elements Chemistry Laboratory, RIKEN Cluster for Pioneering Research kn-affil= affil-num=6 en-affil=Department of Electrical and Electronic Engineering, Okayama University kn-affil= affil-num=7 en-affil=Department of Electrical and Electronic Engineering, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=104 cd-vols= no-issue= article-no= start-page=8789 end-page=8799 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200911 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Channel-pore cation selectivity is a major determinant of Bacillus thuringiensis Cry46Ab mosquitocidal activity en-subtitle= kn-subtitle= en-abstract= kn-abstract=Cry46Ab from Bacillus thuringiensis TK-E6 is a new mosquitocidal toxin with an aerolysin-type architecture, and it is expected to be used as a novel bioinsecticide. Cry46Ab acts as a functional pore-forming toxin, and characteristics of the resulting channel pores, including ion selectivity, have been analyzed. However, the relationship between channel-pore ion selectivity and insecticidal activity remains to be elucidated. To clarify the effects of charged amino acid residues on the ion permeability of channel-pores and the resulting insecticidal activity, in the present study, we constructed Cry46Ab mutants in which a charged amino acid residue within a putative transmembrane ƒΐ-hairpin region was replaced with an oppositely charged residue. Bioassays using Culex pipiens mosquito larvae revealed that the mosquitocidal activity was altered by the mutation. A K155E Cry46Ab mutant exhibited toxicity apparently higher than that of wild-type Cry46Ab, but the E159K and E163K mutants exhibited decreased toxicity. Ions selectivity measurements demonstrated that the channel pores formed by both wild-type and mutant Cry46Abs were cation selective, and their cation preference was also similar. However, the degree of cation selectivity was apparently higher in channel pores formed by the K155E mutant, and reduced selectivity was observed with the E159K and E163K mutants. Our data suggest that channel-pore cation selectivity is a major determinant of Cry46Ab mosquitocidal activity and that cation selectivity can be controlled via mutagenesis targeting the transmembrane ƒΐ-hairpin region. en-copyright= kn-copyright= en-aut-name=HayakawaTohru en-aut-sei=Hayakawa en-aut-mei=Tohru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MiyazakiMidoka en-aut-sei=Miyazaki en-aut-mei=Midoka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HaradaSyoya en-aut-sei=Harada en-aut-mei=Syoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=AsakuraMami en-aut-sei=Asakura en-aut-mei=Mami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=IdeToru en-aut-sei=Ide en-aut-mei=Toru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= en-keyword=Bacillus thuringiensis TK-E6 kn-keyword=Bacillus thuringiensis TK-E6 en-keyword=Cry46Ab toxin kn-keyword=Cry46Ab toxin en-keyword=Culex pipiens mosquito larvae kn-keyword=Culex pipiens mosquito larvae en-keyword=Site-directed mutagenesis kn-keyword=Site-directed mutagenesis en-keyword=Electrophysiologic analysis kn-keyword=Electrophysiologic analysis END start-ver=1.4 cd-journal=joma no-vol=74 cd-vols= no-issue=4 article-no= start-page=301 end-page=306 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=202008 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Immobility-reducing Effects of Ketamine during the Forced Swim Test on 5-HT1A Receptor Activity in the Medial Prefrontal Cortex in an Intractable Depression Model en-subtitle= kn-subtitle= en-abstract= kn-abstract=Ketamine has been clinically proven to ameliorate depression, including treatment-resistant depression. The detailed mechanism of action of ketamine in treatment-resistant depression remains unclear. We examined the effects of ketamine on the immobility times of adrenocorticotropic hormone (ACTH)-treated rats during the forced swim test, and we explored the mechanism by which ketamine acts in this model. We investigated the neuroanatomical site of action by microinjecting ketamine into the medial prefrontal cortex of rats. A significant reduction of the ratsf immobility during the forced swim test was observed after the intraperitoneal injection of ketamine in both saline- and ACTH-treated rats. The microinjection of ketamine into the medial prefrontal cortex also decreased immobility during the forced swim test in both saline- and ACTH-treated rats. The immobility-decreasing effect of intraperitoneally injected ketamine was blocked by administering WAY100635, a 5-HT1A receptor antagonist, into the medial prefrontal cortex. These findings contribute to the evidence that ketamine can be useful against treatment-resistant depressive conditions. The immobility-reducing effects of ketamine might be mediated by 5-HT1A receptor activity in the medial prefrontal cortex. en-copyright= kn-copyright= en-aut-name=TakahashiKei en-aut-sei=Takahashi en-aut-mei=Kei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KitamuraYoshihisa en-aut-sei=Kitamura en-aut-mei=Yoshihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=UshioSoichiro en-aut-sei=Ushio en-aut-mei=Soichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SendoToshiaki en-aut-sei=Sendo en-aut-mei=Toshiaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil=Department of Clinical Pharmacy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Clinical Pharmacy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Clinical Pharmacy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Clinical Pharmacy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=ketamine kn-keyword=ketamine en-keyword=adrenocorticotropic hormone kn-keyword=adrenocorticotropic hormone en-keyword=forced swim test kn-keyword=forced swim test en-keyword=medial prefrontal cortex kn-keyword=medial prefrontal cortex en-keyword=5-HT1A receptor kn-keyword=5-HT1A receptor END start-ver=1.4 cd-journal=joma no-vol=34 cd-vols= no-issue= article-no= start-page=575 end-page=582 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200608 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Improvement of biodistribution profile of a radiogallium-labeled, ƒΏvƒΐ6 integrin-targeting peptide probe by incorporation of negatively charged amino acids en-subtitle= kn-subtitle= en-abstract= kn-abstract=Objective
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers. Since ƒΏvƒΐ6 integrin has been reported as a promising target for PDAC diagnosis, we previously developed H-Cys(mal-NOTA-67Ga)-(Gly)6-A20FMDV2-NH2 ([67Ga]CG6) as an ƒΏvƒΐ6 integrin-targeting probe. Although [67Ga]CG6 specifically binds to ƒΏvƒΐ6 integrin-positive xenografts, the uptake of [67Ga]CG6 in the organs surrounding the pancreas, such as the liver and spleen, was comparable to that in the ƒΏvƒΐ6 integrin-positive xenografts. We hypothesized that the undesirable accumulation of [67Ga]CG6 in those organs was caused by the positive charges of [67Ga]CG6 (+?3). In this study, we aimed to decrease [67Ga]CG6 uptake in the liver and spleen by reducing the electric charges of the probe.
Methods
We synthesized H-Cys(mal-NOTA-67Ga)-(Asp)6-A20FMDV2-NH2 ([67Ga]CD6) and evaluated its affinity to ƒΏvƒΐ6 integrin via in vitro competitive binding assay. Isoelectric points of the probes were determined by electrophoresis. Biodistribution study, autoradiography, and immunostaining for ƒΐ6 integrin were conducted using ƒΏvƒΐ6 integrin-positive and negative tumor-bearing mice.
Results
In vitro competitive binding assay showed that the alteration of the linker had a negligible impact on the affinity of [67Ga]CG6 to ƒΏvƒΐ6 integrin. The results of electrophoresis revealed that [67Ga]CG6 was positively charged whereas [67Ga]CD6 was negatively charged. In the biodistribution study, the uptake of [67Ga]CD6 in the ƒΏvƒΐ6 integrin-positive xenografts was significantly higher than that in the ƒΏvƒΐ6 integrin-negative ones at 60 and 120 min. The uptake of [67Ga]CD6 in the liver and spleen was more than two-fold lower than that of [67Ga]CG6 at both time points. In the immunohistochemistry study, the radioactivity accumulated areas in the autoradiogram of the ƒΏvƒΐ6 integrin-positive xenograft roughly coincided with ƒΐ6 integrin-expressing areas.
Conclusion
We have successfully reduced the nonspecific uptake in the liver and spleen by altering the linker amino acid from G6 to D6. [67Ga]CD6 overcame the drawbacks of [67Ga]CG6 in its biodistribution. en-copyright= kn-copyright= en-aut-name=NakamuraShunsuke en-aut-sei=Nakamura en-aut-mei=Shunsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsunoAya en-aut-sei=Matsuno en-aut-mei=Aya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=UedaMasashi en-aut-sei=Ueda en-aut-mei=Masashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil= Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=ƒΏvƒΐ6 integrin kn-keyword=ƒΏvƒΐ6 integrin en-keyword=Pancreatic ductal adenocarcinoma (PDAC) kn-keyword=Pancreatic ductal adenocarcinoma (PDAC) en-keyword=A20FMDV2 kn-keyword=A20FMDV2 en-keyword=Aspartic acids kn-keyword=Aspartic acids en-keyword=Electric charge kn-keyword=Electric charge END start-ver=1.4 cd-journal=joma no-vol=74 cd-vols= no-issue=3 article-no= start-page=199 end-page=208 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=202006 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Dkk3/REIC, an N-glycosylated Protein, Is a Physiological Endoplasmic Reticulum Stress Inducer in the Mouse Adrenal Gland en-subtitle= kn-subtitle= en-abstract= kn-abstract=Dickkopf 3 (Dkk3) is a secreted protein belonging to the Dkk family and encoded by the orthologous gene of REIC. Dkk3/REIC is expressed by mouse and human adrenal glands, but the understanding of its roles in this organ is still limited. To determine the functions of Dkk3 in the mouse adrenal gland, we first identified that the mouse Dkk3 protein is N-glycosylated in the adrenal gland as well as in the brain. We performed proteome analysis on adrenal glands from Dkk3-null mice, in which exons 5 and 6 of the Dkk3 gene are deleted. Twodimensional polyacrylamide gel electrophoresis of adrenal proteins from wild-type and Dkk3-null mice revealed 5 protein spots whose intensities were altered between the 2 genotypes. Mass spectrometry analysis of these spots identified binding immunoglobulin protein (BiP), an endoplasmic reticulum (ER) chaperone. To determine whether mouse Dkk3 is involved in the unfolded protein response (UPR), we carried out a reporter assay using ER-stress responsive elements. Forced expression of Dkk3 resulted in the induction of distinct levels of reporter expression, showing the UPR initiated by the ER membrane proteins of activating transcription factor 6 (ATF6) and inositol-requring enzyme 1 (IRE1). Thus, it is possible that Dkk3 is a physiological ER stressor in the mouse adrenal gland. en-copyright= kn-copyright= en-aut-name=FujitaHirofumi en-aut-sei=Fujita en-aut-mei=Hirofumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=BandoTetsuya en-aut-sei=Bando en-aut-mei=Tetsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OyadomariSeiichi en-aut-sei=Oyadomari en-aut-mei=Seiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OchiaiKazuhiko en-aut-sei=Ochiai en-aut-mei=Kazuhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=WatanabeMasami en-aut-sei=Watanabe en-aut-mei=Masami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KumonHiromi en-aut-sei=Kumon en-aut-mei=Hiromi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=OhuchiHideyo en-aut-sei=Ohuchi en-aut-mei=Hideyo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Division of Molecular Biology, Institute for Genome Research, University of Tokushima kn-affil= affil-num=4 en-affil=Department of Basic Science, School of Veterinary Nursing and Technology, Faculty of Veterinary Science, Nippon Veterinary and Life Science University kn-affil= affil-num=5 en-affil=Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Innovation Center Okayama for Nanobio-Targeted Therapy, Okayama University kn-affil= affil-num=7 en-affil=Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=Dkk3 knockout mouse kn-keyword=Dkk3 knockout mouse en-keyword=adrenal gland kn-keyword=adrenal gland en-keyword=glucose-regulated protein 78 kn-keyword=glucose-regulated protein 78 en-keyword=proteome kn-keyword=proteome en-keyword=endoplasmic reticulum stress kn-keyword=endoplasmic reticulum stress END start-ver=1.4 cd-journal=joma no-vol=9 cd-vols= no-issue=3 article-no= start-page=643 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200306 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=HMGB1 Translocation in Neurons after Ischemic Insult: Subcellular Localization in Mitochondria and Peroxisomes en-subtitle= kn-subtitle= en-abstract= kn-abstract=High mobility group box-1 (HMGB1), a nonhistone chromatin DNA-binding protein, is released from neurons into the extracellular space under ischemic, hemorrhagic, and traumatic insults. However, the details of the time-dependent translocation of HMGB1 and the subcellular localization of HMGB1 through the release process in neurons remain unclear. In the present study, we examined the subcellular localization of HMGB1 during translocation of HMGB1 in the cytosolic compartment using a middle cerebral artery occlusion and reperfusion model in rats. Double immunofluorescence microscopy revealed that HMGB1 immunoreactivities were colocalized with MTCO1(mitochondrially encoded cytochrome c oxidase I), a marker of mitochondria, and catalase, a marker of peroxisomes, but not with Rab5/Rab7 (RAS-related GTP-binding protein), LC3A/B (microtubule-associated protein 1 light chain 3), KDEL (KDEL amino acid sequence), and LAMP1 (Lysosomal Associated Membrane Protein 1), which are endosome, phagosome, endoplasmic reticulum, and lysosome markers, respectively. Immunoelectron microscopy confirmed that immune-gold particles for HMGB1 were present inside the mitochondria and peroxisomes. Moreover, HMGB1 was found to be colocalized with Drp1 (Dynamin-related protein 1), which is involved in mitochondrial fission. These results revealed the specific subcellular localization of HMGB1 during its release process under ischemic conditions. en-copyright= kn-copyright= en-aut-name=WangDengli en-aut-sei=Wang en-aut-mei=Dengli kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=LiuKeyue en-aut-sei=Liu en-aut-mei=Keyue kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=FukuyasuYusuke en-aut-sei=Fukuyasu en-aut-mei=Yusuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TeshigawaraKiyoshi en-aut-sei=Teshigawara en-aut-mei=Kiyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=FuLi en-aut-sei=Fu en-aut-mei=Li kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=WakeHidenori en-aut-sei=Wake en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=OhtsukaAiji en-aut-sei=Ohtsuka en-aut-mei=Aiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=NishiboriMasahiro en-aut-sei=Nishibori en-aut-mei=Masahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=6 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=7 en-affil=Department of Human Morphology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=8 en-affil=Department of Pharmacology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=middle cerebral artery occlusion kn-keyword=middle cerebral artery occlusion en-keyword=high-mobility group box 1 kn-keyword=high-mobility group box 1 en-keyword=subcellular localization and subcellular organelle kn-keyword=subcellular localization and subcellular organelle END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue=2 article-no= start-page=121 end-page=132 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200514 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The expression level and cytotoxicity of green fluorescent protein are modulated by an additional N-terminal sequence en-subtitle= kn-subtitle= en-abstract= kn-abstract=Nucleotide and amino acid sequences at the N-terminus affect the expression level and cytotoxicity of proteins; however, their effects are not fully understood yet. Here, N-terminal 30 nucleotide/10 amino acid (N10) sequences that affect the expression level and cytotoxicity of a green fluorescent protein were systematically isolated in the budding yeast Saccharomyces cerevisiae. The expression per gene (EPG) and gene copy number limit (CNL) relationships were examined to assess the effects of the N10 sequence. The isolated N10 nucleotide sequences suggested that codon optimality is the major determinant of the protein expression level. A higher number of hydrophobic or cysteine residues in the N10 sequence seemed to increase the cytotoxicity of the protein. Therefore, a high frequency of specific amino acid residues in the outside of the main tertiary structure of proteins might not be preferable. en-copyright= kn-copyright= en-aut-name=MoriyaHisao en-aut-sei=Moriya en-aut-mei=Hisao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Sciences, Research Core for Interdisciplinary Sciences, Okayama University kn-affil= en-keyword=green fluorescent protein kn-keyword=green fluorescent protein en-keyword=overexpression kn-keyword=overexpression en-keyword=expression limit kn-keyword=expression limit en-keyword=expression level kn-keyword=expression level en-keyword=protein cytotoxicity kn-keyword=protein cytotoxicity END start-ver=1.4 cd-journal=joma no-vol=16 cd-vols= no-issue=4 article-no= start-page=e1008469 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200423 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Non-pathogenic Escherichia coli acquires virulence by mutating a growth-essential LPS transporter en-subtitle= kn-subtitle= en-abstract= kn-abstract=The molecular mechanisms that allow pathogenic bacteria to infect animals have been intensively studied. On the other hand, the molecular mechanisms by which bacteria acquire virulence functions are not fully understood. In the present study, we experimentally evaluated the evolution of a non-pathogenic strain of Escherichia coli in a silkworm infection model and obtained pathogenic mutant strains. As one cause of the high virulence properties of E. coli mutants, we identified amino acid substitutions in LptD (G580S) and LptE (T95I) constituting the lipopolysaccharide (LPS) transporter, which translocates LPS from the inner to the outer membrane and is essential for E. coli growth. The growth of the LptD and LptE mutants obtained in this study was indistinguishable from that of the parent strain. The LptD and LptE mutants exhibited increased secretion of outer membrane vesicles containing LPS and resistance against various antibiotics, antimicrobial peptides, and host complement. In vivo cross-linking studies revealed that the conformation of the LptD-LptE complex was altered in the LptD and LptE mutants. Furthermore, several clinical isolates of E. coli carried amino acid substitutions of LptD and LptE that conferred resistance against antimicrobial substances. This study demonstrated an experimental evolution of bacterial virulence properties in an animal infection model and identified functional alterations of the growth-essential LPS transporter that led to high bacterial virulence by conferring resistance against antimicrobial substances. These findings suggest that non-pathogenic bacteria can gain virulence traits by changing the functions of essential genes, and provide new insight to bacterial evolution in a host environment. Author summary Pathogenic bacteria developed their virulence properties by changing the functions of various genes after the emergence of the host animals on earth. The types of gene function alterations that confer bacterial virulence properties, however, have remained unclear. We utilized a silkworm infection model to perform an experimental evolution of bacterial virulence activity. From a non-pathogenic strain of Escherichia coli, we obtained a mutant strain that exhibited 500-fold higher virulence than the original strain and identified mutations of the lipopolysaccharide (LPS) transporter, which translocates LPS onto the bacterial surface, as one cause of the high virulence. The mutations changed the structure of the LPS transporter, increased the secretion of outer membrane vesicles, and enabled bacterial survival in the presence of host antimicrobial substances. This mechanism to gain high virulence occurs naturally, as several E. coli clinical isolates carried mutations of the LPS transporter that confer resistance against antimicrobial substances. Our study unveiled a novel mechanism by which bacteria increase their virulence through modifying their gene function. en-copyright= kn-copyright= en-aut-name=KaitoChikara en-aut-sei=Kaito en-aut-mei=Chikara kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YoshikaiHirono en-aut-sei=Yoshikai en-aut-mei=Hirono kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WakamatsuAi en-aut-sei=Wakamatsu en-aut-mei=Ai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MiyashitaAtsushi en-aut-sei=Miyashita en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MatsumotoYasuhiko en-aut-sei=Matsumoto en-aut-mei=Yasuhiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=FujiyukiTomoko en-aut-sei=Fujiyuki en-aut-mei=Tomoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KatoMasaru en-aut-sei=Kato en-aut-mei=Masaru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=OguraYoshitoshi en-aut-sei=Ogura en-aut-mei=Yoshitoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=HayashiTetsuya en-aut-sei=Hayashi en-aut-mei=Tetsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=IsogaiTakao en-aut-sei=Isogai en-aut-mei=Takao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=SekimizuKazuhisa en-aut-sei=Sekimizu en-aut-mei=Kazuhisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Pharmaceutical Sciences, The University of Tokyo kn-affil= affil-num=3 en-affil=Japan Biological Informatics Consortium (JBIC) kn-affil= affil-num=4 en-affil=Graduate School of Pharmaceutical Sciences, The University of Tokyo kn-affil= affil-num=5 en-affil=Department of Microbiology, Meiji Pharmaceutical University kn-affil= affil-num=6 en-affil=The Institute of Medical Science, The University of Tokyo kn-affil= affil-num=7 en-affil=Devision of Bioanalytical Chemistry, School of Pharmacy,Showa University kn-affil= affil-num=8 en-affil=Department of Bacteriology, Faculty of Medical Sciences,Kyushu University kn-affil= affil-num=9 en-affil=Department of Bacteriology, Faculty of Medical Sciences,Kyushu University kn-affil= affil-num=10 en-affil=Translational Research Center, Fukushima Medical University kn-affil= affil-num=11 en-affil=Institute of Medical Mycology, Teikyo University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=14 cd-vols= no-issue=10 article-no= start-page=e0218909 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=20191004 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Differential scanning fluorimetric analysis of the amino-acid binding to taste receptor using a model receptor protein, the ligand-binding domain of fish T1r2a/T1r3 en-subtitle= kn-subtitle= en-abstract= kn-abstract=Taste receptor type 1 (T1r) is responsible for the perception of essential nutrients, such as sugars and amino acids, and evoking sweet and umami (savory) taste sensations. T1r receptors recognize many of the taste substances at their extracellular ligand-binding domains (LBDs). In order to detect a wide array of taste substances in the environment, T1r receptors often possess broad ligand specificities. However, the entire ranges of chemical spaces and their binding characteristics to any T1rLBDs have not been extensively analyzed. In this study, we exploited the differential scanning fluorimetry (DSF) to medaka T1r2a/T1r3LBD, a current sole T1rLBD heterodimer amenable for recombinant preparation, and analyzed their thermal stabilization by adding various amino acids. The assay showed that the agonist amino acids induced thermal stabilization and shifted the melting temperatures (T-m) of the protein. An agreement between the DSF results and the previous biophysical assay was observed, suggesting that DSF can detect ligand binding at the orthostericbinding site in T1r2a/T1r3LBD. The assay further demonstrated that most of the tested Lamino acids, but no D-amino acid, induced T-m shifts of T1r2a/T1r3LBD, indicating the broad L-amino acid specificities of the proteins probably with several different manners of recognition. The T-m shifts by each amino acid also showed a fair correlation with the responses exhibited by the full-length receptor, verifying the broad amino-acid binding profiles at the orthosteric site in LBD observed by DSF. en-copyright= kn-copyright= en-aut-name=YoshidaTakashi en-aut-sei=Yoshida en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YasuiNorihisa en-aut-sei=Yasui en-aut-mei=Norihisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KusakabeYuko en-aut-sei=Kusakabe en-aut-mei=Yuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ItoChiaki en-aut-sei=Ito en-aut-mei=Chiaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=AkamatsuMiki en-aut-sei=Akamatsu en-aut-mei=Miki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=YamashitaAtsuko en-aut-sei=Yamashita en-aut-mei=Atsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Food Research Institute, National Agriculture and Food Research Organization kn-affil= affil-num=4 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Agriculture, Kyoto University kn-affil= affil-num=6 en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page=135041 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200513 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effects of Bitter Receptor Antagonists on Behavioral Lick Responses of Mice en-subtitle= kn-subtitle= en-abstract= kn-abstract= Bitter taste receptors TAS2Rs detect noxious compounds in the oral cavity. Recent heterologous expression studies reported that some compounds function as antagonists for human TAS2Rs. For examples, amino acid derivatives such as ƒΑ-aminobutyric acid (GABA) and NƒΏ,NƒΏ-bis(carboxymethyl)-L-Lysine (BCML) blocked responses to quinine mediated by human TAS2R4. Probenecid inhibited responses to phenylthiocarbamide mediated by human TAS2R38. In this study, we investigated the effects of these human bitter receptor antagonists on behavioral lick responses of mice to elucidate whether these compounds also function as bitter taste blockers. In short-term (10 s) lick tests, concentration-dependent lick responses to bitter compounds (quinine-HCl, denatonium and phenylthiourea) were not affected by the addition of GABA or BCML. Probenecid reduced aversive lick responses to denatonium and phenylthiourea but not to quinine-HCl. In addition, taste cell responses to phenylthiourea were inhibited by probenecid. These results suggest some bitter antagonists of human TAS2Rs can work for bitter sense of mouse. en-copyright= kn-copyright= en-aut-name=MasamotoMichimasa en-aut-sei=Masamoto en-aut-mei=Michimasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MitohYoshihiro en-aut-sei=Mitoh en-aut-mei=Yoshihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KobashiMotoi en-aut-sei=Kobashi en-aut-mei=Motoi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ShigemuraNoriatsu en-aut-sei=Shigemura en-aut-mei=Noriatsu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YoshidaRyusuke en-aut-sei=Yoshida en-aut-mei=Ryusuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=3 en-affil=Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= affil-num=4 en-affil=Section of Oral Neuroscience, Graduate School of Dental Sciences, Kyushu University kn-affil= affil-num=5 en-affil=Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University kn-affil= en-keyword=bitter coding kn-keyword=bitter coding en-keyword=bitter inhibitor kn-keyword=bitter inhibitor en-keyword=gustatory response kn-keyword=gustatory response en-keyword=species difference kn-keyword=species difference en-keyword=taste perception kn-keyword=taste perception END start-ver=1.4 cd-journal=joma no-vol=223-225 cd-vols= no-issue= article-no= start-page=72 end-page=78 dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=201908 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Quorum-dependent expression of rsmX and rsmY, small non-coding RNAs, in Pseudomonas syringae en-subtitle= kn-subtitle= en-abstract= kn-abstract=Pseudomonas syringae pathovars are known to produce N-acyl-homoserine lactones (AHL) as quorum-sensing molecules. However, many isolates, including P. syringae pv. tomato DC3000 (PtoDC3000), do not produce them. In P. syringae, psyI, which encodes an AHL synthase, and psyR, which encodes the transcription factor PsyR required for activation of psyI, are convergently transcribed. In P. amygdali pv. tabaci 6605 (Pta6605), there is one nucleotide between the stop codons of both psyI and psyR. However, the canonical stop codon for psyI in PtoDC3000 was converted to the cysteine codon by one nucleotide deletion, and 23 additional amino acids extended it to a C-terminal end. This resulted in overlapping of the open reading frame (ORF) for psyI and psyR. On the other hand, stop codons in the psyR ORF of P. syringae 7 isolates, including pv. phaseolicola and pv. glycinea, were found. These results indicate that many pathovars of P. syringae have genetically lost AHL production ability by the mutation of their responsible genes. To examine whether PtoDC3000 modulates the gene expression profile in a population-dependent manner, we carried out microarray analysis using RNAs prepared from low- and high-density cells. We found the expressions of rsmX and rsmY remarkably activated in high-density cells. The activated expressions of rsmX and rsmY were confirmed by Northern blot hybridization, but these expressions were abolished in a ƒ’gacA mutant of Pta6605. These results indicate that regardless of the ability to produce AHL, P. syringae regulates expression of the small noncoding RNAs rsmX/Y by currently unknown quorum-sensing molecules. en-copyright= kn-copyright= en-aut-name=NakatsuYukiko en-aut-sei=Nakatsu en-aut-mei=Yukiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsuiHidenori en-aut-sei=Matsui en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamamotoMikihiro en-aut-sei=Yamamoto en-aut-mei=Mikihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=NoutoshiYoshiteru en-aut-sei=Noutoshi en-aut-mei=Yoshiteru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ToyodaKazuhiro en-aut-sei=Toyoda en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=2 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=3 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=4 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=5 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=6 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= en-keyword=N-acyl-homoserine lactone kn-keyword=N-acyl-homoserine lactone en-keyword=Gac two-component system kn-keyword=Gac two-component system en-keyword=Quorum sensing kn-keyword=Quorum sensing en-keyword=rsmX kn-keyword=rsmX en-keyword=rsmY kn-keyword=rsmY en-keyword=Pseudomonas syringae kn-keyword=Pseudomonas syringae END start-ver=1.4 cd-journal=joma no-vol=24 cd-vols= no-issue=8 article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=20190416 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Melittin Exerts Beneficial Effects on Paraquat-Induced Lung Injuries in Mice by Modifying Oxidative Stress and Apoptosis en-subtitle= kn-subtitle= en-abstract= kn-abstract=Melittin (MEL) is a 26-amino acid peptide with numerous biological activities. Paraquat (PQ) is one of the most widely used herbicides, although it is extremely toxic to humans. To date, PQ poisoning has no effective treatment, and therefore the current study aimed to assess for the first time the possible effects of MEL on PQ-induced lung injuries in mice. Mice received a single intraperitoneal (IP) injection of PQ (30 mg/kg), followed by IP treatment with MEL (0.1 and 0.5 mg/kg) twice per week for four consecutive weeks. Histological alterations, oxidative stress, and apoptosis in the lungs were studied. Hematoxylin and eosin (H&E) staining indicated that MEL markedly reduced lung injuries induced by PQ. Furthermore, treatment with MEL increased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity, and decreased malonaldehyde (MDA) and nitric oxide (NO) levels in lung tissue homogenates. Moreover, immunohistochemical staining showed that B-cell lymphoma-2 (Bcl-2) and survivin expressions were upregulated after MEL treatment, while Ki-67 expression was downregulated. The high dose of MEL was more effective than the low dose in all experiments. In summary, MEL efficiently reduced PQ-induced lung injuries in mice. Specific pharmacological examinations are required to determine the effectiveness of MEL in cases of human PQ poisoning. en-copyright= kn-copyright= en-aut-name=El-AaragBishoy en-aut-sei=El-Aarag en-aut-mei=Bishoy kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MagdyMohamed en-aut-sei=Magdy en-aut-mei=Mohamed kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AlAjmiMohamed F. en-aut-sei=AlAjmi en-aut-mei=Mohamed F. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KhalifaShaden A. M. en-aut-sei=Khalifa en-aut-mei=Shaden A. M. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=El-SeediHesham R. en-aut-sei=El-Seedi en-aut-mei=Hesham R. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Division of Chemistry and Biotechnology, Graduate School of Natural Science and Technology, Okayama University kn-affil= affil-num=2 en-affil=Department of Chemistry, Faculty of Science, Menoufia University kn-affil= affil-num=3 en-affil=Department of Pharmacognosy, College of Pharmacy, King Saud University kn-affil= affil-num=4 en-affil=Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University kn-affil= affil-num=5 en-affil=Department of Chemistry, Faculty of Science, Menoufia University kn-affil= en-keyword=paraquat kn-keyword=paraquat en-keyword=lung injury kn-keyword=lung injury en-keyword=melittin kn-keyword=melittin en-keyword=oxidative stress kn-keyword=oxidative stress en-keyword=apoptosis kn-keyword=apoptosis END start-ver=1.4 cd-journal=joma no-vol=74 cd-vols= no-issue=1 article-no= start-page=65 end-page=72 dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=202002 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Metabolic Profiling of the Cerebrospinal Fluid in Pediatric Epilepsy en-subtitle= kn-subtitle= en-abstract= kn-abstract= To characterize metabolic profiles within the central nervous system in epilepsy, we performed gas chromatography-tandem mass spectrometry (GC-MS/MS)-based metabolome analysis of the cerebrospinal fluid (CSF) in pediatric patients with and without epilepsy. The CSF samples obtained from 64 patients were analyzed by GC-MS/MS. Multivariate analyses were performed for two age groups, 0-5 years of age and 6-17 years of age, to elucidate the effects of epilepsy and antiepileptic drugs on the metabolites. In patients aged 0-5 years (22 patients with epilepsy, 13 without epilepsy), epilepsy patients had reduced 2-ketoglutaric acid and elevated pyridoxamine and tyrosine. In patients aged 6-17 years (12 with epilepsy, 17 without epilepsy), epilepsy patients had reduced 1,5-anhydroglucitol. Valproic acid was associated with elevated 2-aminobutyric acid, 2-ketoisocaproic acid, 4-hydroxyproline, acetylglycine, methionine, N-acetylserine, and serine. Reduced energy metabolism and alteration of vitamin B6 metabolism may play a role in epilepsy in young children. The roles of 1,5-anhydroglucitol in epilepsy in older children and in levetiracetam and zonisamide treatment remain to be explained. Valproic acid influenced the levels of amino acids and related metabolites involved in the metabolism of serine, methionine, and leucine. en-copyright= kn-copyright= en-aut-name=AkiyamaTomoyuki en-aut-sei=Akiyama en-aut-mei=Tomoyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SaigusaDaisuke en-aut-sei=Saigusa en-aut-mei=Daisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HyodoYuki en-aut-sei=Hyodo en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=UmedaKeiko en-aut-sei=Umeda en-aut-mei=Keiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SaijoReina en-aut-sei=Saijo en-aut-mei=Reina kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KoshibaSeizo en-aut-sei=Koshiba en-aut-mei=Seizo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KobayashiKatsuhiro en-aut-sei=Kobayashi en-aut-mei=Katsuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= affil-num=2 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=3 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= affil-num=4 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=5 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=6 en-affil=Tohoku Medical Megabank Organization, Tohoku University kn-affil= affil-num=7 en-affil=Department of Child Neurology, Okayama University Hospital kn-affil= en-keyword=antiepileptic drugs kn-keyword=antiepileptic drugs en-keyword=gas chromatography-tandem mass spectrometry kn-keyword=gas chromatography-tandem mass spectrometry en-keyword=metabolome analysis kn-keyword=metabolome analysis en-keyword=metabolomics kn-keyword=metabolomics END start-ver=1.4 cd-journal=joma no-vol=159 cd-vols= no-issue=1 article-no= start-page=163 end-page=166 dt-received= dt-revised= dt-accepted= dt-pub-year=2013 dt-pub=20130716 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Complete genome sequence of Habenaria mosaic virus, a new potyvirus infecting a terrestrial orchid (Habenaria radiata) in Japan en-subtitle= kn-subtitle= en-abstract= kn-abstract= The complete genomic sequence of Habenaria mosaic virus (HaMV), which infects terrestrial orchids (Habenaria radiata), has been determined. The genome is composed of 9,499 nucleotides excluding the 3'-terminal poly(A) tail, encoding a large polyprotein of 3,054 amino acids with the genomic features typical of a potyvirus. Putative proteolytic cleavage sites were identified by sequence comparison to those of known potyviruses. The HaMV polyprotein showed 58 % amino acid sequence identity to that encoded by the most closely related potyvirus, tobacco vein banding mosaic virus. Phylogenetic analysis of the polyprotein amino acid sequence and its coding sequences confirmed that HaMV formed a cluster with the chilli veinal mottle virus group, most of which infect solanaceous plants. These results suggest that HaMV is a distinct member of the genus Potyvirus. en-copyright= kn-copyright= en-aut-name=KondoHideki en-aut-sei=Kondo en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MaedaTakanori en-aut-sei=Maeda en-aut-mei=Takanori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=I Wayan Gara en-aut-sei=I Wayan Gara en-aut-mei= kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ChibaSotaro en-aut-sei=Chiba en-aut-mei=Sotaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MaruyamaKazuyuki en-aut-sei=Maruyama en-aut-mei=Kazuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TamadaTetsuo en-aut-sei=Tamada en-aut-mei=Tetsuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SuzukiNobuhiro en-aut-sei=Suzuki en-aut-mei=Nobuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=2 en-affil=College of Bioresource SciencesNihon University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources (IPSR)Okayama University kn-affil= affil-num=4 en-affil=Institute of Plant Science and Resources (IPSR)Okayama University kn-affil= affil-num=5 en-affil=Institute of Plant Science and Resources (IPSR)Okayama University kn-affil= affil-num=6 en-affil=Institute of Plant Science and Resources (IPSR)Okayama University kn-affil= affil-num=7 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=36 cd-vols= no-issue=18 article-no= start-page=1359 end-page=1369 dt-received= dt-revised= dt-accepted= dt-pub-year=2015 dt-pub=20150531 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A solvation-free-energy functional: A reference-modified density functional formulation en-subtitle= kn-subtitle= en-abstract= kn-abstract= The three-dimensional reference interaction site model (3D-RISM) theory, which is one of the most applicable integral equation theories for molecular liquids, overestimates the absolute values of solvation-free-energy (SFE) for large solute molecules in water. To improve the free-energy density functional for the SFE of solute molecules, we propose a reference-modified density functional theory (RMDFT) that is a general theoretical approach to construct the free-energy density functional systematically. In the RMDFT formulation, hard-sphere (HS) fluids are introduced as the reference system instead of an ideal polyatomic molecular gas, which has been regarded as the appropriate reference system of the interaction-site-model density functional theory for polyatomic molecular fluids. We show that using RMDFT with a reference HS system can significantly improve the absolute values of the SFE for a set of neutral amino acid side-chain analogues as well as for 504 small organic molecules. en-copyright= kn-copyright= en-aut-name=SumiTomonari en-aut-sei=Sumi en-aut-mei=Tomonari kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MitsutakeAyori en-aut-sei=Mitsutake en-aut-mei=Ayori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MaruyamaYutaka en-aut-sei=Maruyama en-aut-mei=Yutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=Department of Chemistry, Faculty of Science, Okayama University, kn-affil= affil-num=2 en-affil=Department of Physics, Keio University, kn-affil= affil-num=3 en-affil=Department of Physics, Keio University, kn-affil= en-keyword=salvation-free-energy kn-keyword=salvation-free-energy en-keyword=classical density functional theory kn-keyword=classical density functional theory en-keyword=3D-RISM theory kn-keyword=3D-RISM theory en-keyword= water kn-keyword= water en-keyword=amino acid side-chain kn-keyword=amino acid side-chain en-keyword=chignolin kn-keyword=chignolin END start-ver=1.4 cd-journal=joma no-vol=213 cd-vols= no-issue= article-no= start-page=353 end-page=364 dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=201602 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Sequence and phylogenetic analyses of novel totivirus-like double-stranded RNAs from field-collected powdery mildew fungi en-subtitle= kn-subtitle= en-abstract= kn-abstract= The identification of mycoviruses contributes greatly to understanding of the diversity and evolutionary aspects of viruses. Powdery mildew fungi are important and widely studied obligate phytopathogenic agents, but there has been no report on mycoviruses infecting these fungi. In this study, we used a deep sequencing approach to analyze the double-stranded RNA (dsRNA) segments isolated from field-collected samples of powdery mildew fungus-infected red clover plants in Japan. Database searches identified the presence of at least ten totivirus (genus Totivirus)-like sequences, termed red clover powdery mildew-associated totiviruses (RPaTVs). The majority of these sequences shared moderate amino acid sequence identity with each other (<44%) and with other known totiviruses (<59%). Nine of these identified sequences (RPaTV1a, 1b and 2-8) resembled the genome of the prototype totivirus, Saccharomyces cerevisiae virus-L-A (ScV-L-A) in that they contained two overlapping open reading frames (ORFs) encoding a putative coat protein (CP) and an RNA dependent RNA polymerase (RdRp), while one sequence (RPaTV9) showed similarity to another totivirus, Ustilago maydis virus H1 (UmV-H1) that encodes a single polyprotein (CP-RdRp fusion). Similar to yeast totiviruses, each ScV-L-A-like RPaTV contains a -1 ribosomal frameshift site downstream of a predicted pseudoknot structure in the overlapping region of these ORFs, suggesting that the RdRp is translated as a CP-RdRp fusion. Moreover, several ScV-L-A-like sequences were also found by searches of the transcriptome shotgun assembly (TSA) libraries from rust fungi, plants and insects. Phylogenetic analyses show that nine ScV-L-A-like RPaTVs along with ScV-L-A-like sequences derived from TSA libraries are clustered with most established members of the genus Totivirus, while one RPaTV forms a new distinct clade with UmV-H1, possibly establishing an additional genus in the family. Taken together, our results indicate the presence of diverse, novel totiviruses in the powdery mildew fungus populations infecting red clover plants in the field. en-copyright= kn-copyright= en-aut-name=KondoHideki en-aut-sei=Kondo en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HisanoSakae en-aut-sei=Hisano en-aut-mei=Sakae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=ChibaSotaro en-aut-sei=Chiba en-aut-mei=Sotaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MaruyamaKazuyuki en-aut-sei=Maruyama en-aut-mei=Kazuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=AndikaIda Bagus en-aut-sei=Andika en-aut-mei=Ida Bagus kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=ToyodaKazuhiro en-aut-sei=Toyoda en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=FujimoriFumihiro en-aut-sei=Fujimori en-aut-mei=Fumihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=SuzukiNobuhiro en-aut-sei=Suzuki en-aut-mei=Nobuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=4 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=5 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= affil-num=6 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil= affil-num=7 en-affil=Department of Environmental Education, Tokyo Kasei University kn-affil= affil-num=8 en-affil=Institute of Plant Science and Resources (IPSR), Okayama University kn-affil= en-keyword=Deep sequencing kn-keyword=Deep sequencing en-keyword=Double stranded RNA virus kn-keyword=Double stranded RNA virus en-keyword= Powdery mildew kn-keyword= Powdery mildew en-keyword=Saccharomyces cerevisiae virus L-A kn-keyword=Saccharomyces cerevisiae virus L-A en-keyword=Totivirus kn-keyword=Totivirus en-keyword=Ustilago maydis virus H1 kn-keyword=Ustilago maydis virus H1 END start-ver=1.4 cd-journal=joma no-vol=73 cd-vols= no-issue=5 article-no= start-page=379 end-page=382 dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=201910 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Histidine-rich Glycoprotein Modulates the Blood-vascular System in Septic Condition en-subtitle= kn-subtitle= en-abstract= kn-abstract= Histidine-rich glycoprotein (HRG) is a 75 kDa glycoprotein synthesized in the liver whose plasma concentration is 100-150 ƒΚg/ml. HRG has been shown to modulate sepsis-related biological reactions by binding to several substances and cells, including heparin, factor XII, fibrinogen, thrombospondin, plasminogen, C1q, IgG, heme, LPS, dead cells, bacteria, and fungi. Therefore, reduction of plasma HRG levels in sepsis leads to dysregulation of coagulation, fibrinolysis, and immune response, resulting in disseminated intravascular coagulation and multiple organ failure. This review summarizes the binding and functional properties of HRG in sepsis. en-copyright= kn-copyright= en-aut-name=WakeHidenori en-aut-sei=Wake en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil=Department of Pharmacology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences kn-affil= en-keyword=htidine-rich glycoprotein kn-keyword=htidine-rich glycoprotein en-keyword=septic pathogenesis kn-keyword=septic pathogenesis en-keyword=immunothrombosis kn-keyword=immunothrombosis END start-ver=1.4 cd-journal=joma no-vol=73 cd-vols= no-issue=3 article-no= start-page=189 end-page=195 dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=201906 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Anti-N-Methyl-D-Aspartate Receptor Encephalitis in Psychiatry en-subtitle= kn-subtitle= en-abstract= kn-abstract=Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a recently-discovered autoimmune disorder in which antibodies target NMDAR in the brain. The number of reported cases of anti-NMDAR encephalitis has increased rapidly. Anti-NMDAR encephalitis can be mistakenly diagnosed as psychiatric disorders because many patients present with prominent psychiatric symptoms and visit psychiatric institutions first. Thus, psychiatrists should cultivate a better understanding of anti-NMDAR encephalitis. In this review, we present the mechanisms, epidemiology, symptoms and clinical course, diagnostic tests, treatment and outcomes of patients with anti-NMDAR encephalitis. Furthermore, we discuss the diversity of clinical spectra of anti-NMDAR encephalitis, and demonstrate a differential diagnosis of psychiatric disease from the perspective of psychiatry. en-copyright= kn-copyright= en-aut-name=SakamotoShinji en-aut-sei=Sakamoto en-aut-mei=Shinji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KawaiHiroki en-aut-sei=Kawai en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OkahisaYuko en-aut-sei=Okahisa en-aut-mei=Yuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TsutsuiKo en-aut-sei=Tsutsui en-aut-mei=Ko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KanbayashiTakashi en-aut-sei=Kanbayashi en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TanakaKeiko en-aut-sei=Tanaka en-aut-mei=Keiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MizukiYutaka en-aut-sei=Mizuki en-aut-mei=Yutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TakakiManabu en-aut-sei=Takaki en-aut-mei=Manabu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=YamadaNorihito en-aut-sei=Yamada en-aut-mei=Norihito kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Neuropsychiatry, Akita University Graduate School of Medicine kn-affil= affil-num=5 en-affil=Department of Neuropsychiatry, Akita University Graduate School of Medicine kn-affil= affil-num=6 en-affil=Department of Animal Model Development, Brain Research Institute, Niigata University kn-affil= affil-num=7 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=9 en-affil=Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=NMDAR kn-keyword=NMDAR en-keyword=encephalitis kn-keyword=encephalitis en-keyword=psychiatric symptom kn-keyword=psychiatric symptom en-keyword=schizophrenia kn-keyword=schizophrenia en-keyword=mood disorder kn-keyword=mood disorder END start-ver=1.4 cd-journal=joma no-vol=32 cd-vols= no-issue=supplment 1 article-no= start-page=A20 end-page=A28 dt-received= dt-revised= dt-accepted= dt-pub-year=2014 dt-pub=20140811 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Hospital based surveillance and genetic characterization of rotavirus strains in children (<5 years) with acute gastroenteritis in Kolkata, India, revealed resurgence of G9 and G2 genotypes during 2011-2013 en-subtitle= kn-subtitle= en-abstract= kn-abstract=INTRODUCTION:@ India accounts for an estimated 457,000-884,000 hospitalizations and 2 million outpatient visits for diarrhea. In spite of the huge burden of rotavirus (RV) disease, RV vaccines have not been introduced in national immunization programme of India. Therefore, continuous surveillance for prevalence and monitoring of the circulating genotypes is needed to assess the disease burden prior to introduction of vaccines in this region.@ METHODS:@ During January 2011 through December 2013, 830 and 1000 stool samples were collected from hospitalized and out-patient department (OPD) patients, respectively, in two hospitals in Kolkata, Eastern India. After primary screening, the G-P typing was done by multiplex semi-nested PCR using type specific primers followed by sequencing. Phylogenetic analysis for the VP7 gene of 25 representative strains was done.@ RESULTS:@ Among hospitalized and OPD patients, 53.4% and 47.5% cases were positive for rotaviruses, respectively. Unlike previous studies where G1 was predominant, in hospitalized cases G9 rotavirus strains were most prevalent (40%), followed by G2 (39.6%) whereas G1 and G12 occurred at 16.4% and 5.6% frequency. In OPD cases, the most prevalent strain was G2 (40.3%), followed by G1, G9 and G12 at 25.5%, 22.8%, 9.3%, respectively. Phylogenetically the G1, G2 and G9 strains from Kolkata did not cluster with corresponding genotypes of Rotarix, RotaTeq and Rotavac (116E) vaccine strains.@ CONCLUSION:@ The study highlights the high prevalence of RV in children with gastroenteritis in Kolkata. The circulating genotypes have changed over the time with predominance of G9 and G2 strains during 2011-2013. The current G2, G9 and G1 Kolkata strains shared low amino acid homologies with current vaccine strains. Although there is substantial evidence for cross protection of vaccines against a variety of strains, still the strain variation should be monitored post vaccine introduction to determine if it has any impact on vaccine effectiveness. en-copyright= kn-copyright= en-aut-name=MullickSatarupa en-aut-sei=Mullick en-aut-mei=Satarupa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MandalPaulami en-aut-sei=Mandal en-aut-mei=Paulami kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=Mukti Kant Nayak en-aut-sei=Mukti Kant Nayak en-aut-mei= kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=GhoshSouvik en-aut-sei=Ghosh en-aut-mei=Souvik kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=DePapiya en-aut-sei=De en-aut-mei=Papiya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=RajendranK. en-aut-sei=Rajendran en-aut-mei=K. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=BhattacharyaMihir K. en-aut-sei=Bhattacharya en-aut-mei=Mihir K. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MitraUtpala en-aut-sei=Mitra en-aut-mei=Utpala kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=RamamurthyThandavarayan en-aut-sei=Ramamurthy en-aut-mei=Thandavarayan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=KobayashiNobumichi en-aut-sei=Kobayashi en-aut-mei=Nobumichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=Chawla-SarkarMamta en-aut-sei=Chawla-Sarkar en-aut-mei=Mamta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=2 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=3 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=4 en-affil=Department of Hygiene, Sapporo Medical University School of Medicine kn-affil= affil-num=5 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=6 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=7 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=8 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=9 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= affil-num=10 en-affil=Department of Hygiene, Sapporo Medical University School of Medicine kn-affil= affil-num=11 en-affil=National Institute of Cholera and Enteric Diseases kn-affil= en-keyword=Diarrhea kn-keyword=Diarrhea en-keyword=Rotavirus kn-keyword=Rotavirus en-keyword=India kn-keyword=India en-keyword=Kolkata kn-keyword=Kolkata en-keyword=G9 strains kn-keyword=G9 strains en-keyword=G2 strains kn-keyword=G2 strains END start-ver=1.4 cd-journal=joma no-vol=73 cd-vols= no-issue=1 article-no= start-page=29 end-page=39 dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=201902 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Histidine-rich Glycoprotein Could Be an Early Predictor of Vasospasm after Aneurysmal Subarachnoid Hemorrhage en-subtitle= kn-subtitle= en-abstract= kn-abstract= Cerebral vasospasm (CVS) is a major contributor to the high morbidity and mortality of aneurysmal subarachnoid hemorrhage (aSAH) patients. We measured histidine-rich glycoprotein (HRG), a new biomarker of aSAH, in cerebrospinal fluid (CSF) to investigate whether HRG might be an early predictor of CVS. A total of seven controls and 14 aSAH patients (8 males, 6 females aged 53.4}15.4 years) were enrolled, and serial CSF and serum samples were taken. We allocated these samples to three phases (T1-T3) and measured HRG, interleukin (IL)-6, fibrinopeptide A (FpA), and 8-hydroxy-2f-deoxyguanosine (8OHdG) in the CSF, and the HRG in serum. We also examined the release of HRG in rat blood incubated in artificial CSF. In contrast to the other biomarkers examined, the change in the CSF HRG concentration was significantly different between the nonspasm and spasm groups (p<0.01). The rat blood/CSF model revealed a time course similar to that of the human CSF samples in the non-spasm group. HRG thus appears to have the potential to become an early predictor of CVS. In addition, the interaction of HRG with IL-6, FpA, and 8OHdG may form the pathology of CVS. en-copyright= kn-copyright= en-aut-name=MatsumotoAtsushi en-aut-sei=Matsumoto en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NakamuraTakehiro en-aut-sei=Nakamura en-aut-mei=Takehiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=ShinomiyaAya en-aut-sei=Shinomiya en-aut-mei=Aya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KawakitaKenya en-aut-sei=Kawakita en-aut-mei=Kenya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KawanishiMasahiko en-aut-sei=Kawanishi en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MiyakeKeisuke en-aut-sei=Miyake en-aut-mei=Keisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KurodaYasuhiro en-aut-sei=Kuroda en-aut-mei=Yasuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KeepRichard F. en-aut-sei=Keep en-aut-mei=Richard F. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=TamiyaTakashi en-aut-sei=Tamiya en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Department of Neurological Surgery, Kagawa University Faculty of Medicine kn-affil= affil-num=2 en-affil=Department of Medical Technology, Kagawa Prefectural University of Health Sciences kn-affil= affil-num=3 en-affil=Department of Neurological Surgery, Kagawa University Faculty of Medicine kn-affil= affil-num=4 en-affil=Emergency Medical Center, Kagawa University Hospital kn-affil= affil-num=5 en-affil=Department of Neurological Surgery, Kagawa University Faculty of Medicine kn-affil= affil-num=6 en-affil=Department of Neurological Surgery, Kagawa University Faculty of Medicine kn-affil= affil-num=7 en-affil=Emergency Medical Center, Kagawa University Hospital kn-affil= affil-num=8 en-affil=Department of Neurosurgery, University of Michigan kn-affil= affil-num=9 en-affil=Department of Neurological Surgery, Kagawa University Faculty of Medicine kn-affil= en-keyword=biomarker kn-keyword=biomarker en-keyword=histidine-rich glycoprotein kn-keyword=histidine-rich glycoprotein en-keyword=predictor kn-keyword=predictor en-keyword=subarachnoid hemorrhage kn-keyword=subarachnoid hemorrhage en-keyword=vasospasm kn-keyword=vasospasm END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue= article-no= start-page=e35122 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2018 dt-pub=20180331 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Metabolic co-dependence drives the evolutionarily ancient Hydra-Chlorella symbiosis en-subtitle= kn-subtitle= en-abstract= kn-abstract= Many multicellular organisms rely on symbiotic associations for support of metabolic activity, protection, or energy. Understanding the mechanisms involved in controlling such interactions remains a major challenge. In an unbiased approach we identified key players that control the symbiosis between Hydra viridissima and its photosynthetic symbiont Chlorella sp. A99. We discovered significant up-regulation of Hydra genes encoding a phosphate transporter and glutamine synthetase suggesting regulated nutrition supply between host and symbionts. Interestingly, supplementing the medium with glutamine temporarily supports in vitro growth of the otherwise obligate symbiotic Chlorella, indicating loss of autonomy and dependence on the host. Genome sequencing of Chlorella sp. A99 revealed a large number of amino acid transporters and a degenerated nitrate assimilation pathway, presumably as consequence of the adaptation to the host environment. Our observations portray ancient symbiotic interactions as a codependent partnership in which exchange of nutrients appears to be the primary driving force. en-copyright= kn-copyright= en-aut-name=HamadaMayuko en-aut-sei=Hamada en-aut-mei=Mayuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=Schr?derKatja en-aut-sei=Schr?der en-aut-mei=Katja kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=BathiaJay en-aut-sei=Bathia en-aut-mei=Jay kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=K?rnUlrich en-aut-sei=K?rn en-aut-mei=Ulrich kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=FrauneSebastian en-aut-sei=Fraune en-aut-mei=Sebastian kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KhalturinaMariia en-aut-sei=Khalturina en-aut-mei=Mariia kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KhalturinKonstantin en-aut-sei=Khalturin en-aut-mei=Konstantin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=ShinzatoChuya en-aut-sei=Shinzato en-aut-mei=Chuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=SatohNori en-aut-sei=Satoh en-aut-mei=Nori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=BoschThomas CG en-aut-sei=Bosch en-aut-mei=Thomas CG kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil=Ushimado Marine Institute, Okayama University kn-affil= affil-num=2 en-affil= Interdisciplinary Research Center, Kiel Life Science, Kiel University kn-affil= affil-num=3 en-affil= Interdisciplinary Research Center, Kiel Life Science, Kiel University kn-affil= affil-num=4 en-affil= Interdisciplinary Research Center, Kiel Life Science, Kiel University kn-affil= affil-num=5 en-affil= Interdisciplinary Research Center, Kiel Life Science, Kiel University kn-affil= affil-num=6 en-affil=Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University kn-affil= affil-num=7 en-affil=Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University kn-affil= affil-num=8 en-affil=Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University kn-affil= affil-num=9 en-affil=Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University kn-affil= affil-num=10 en-affil= Interdisciplinary Research Center, Kiel Life Science, Kiel University kn-affil= en-keyword=Chlorella kn-keyword=Chlorella en-keyword=Hydra kn-keyword=Hydra en-keyword=evolutionary biology kn-keyword=evolutionary biology en-keyword=genome kn-keyword=genome en-keyword=nitrogen metabolism kn-keyword=nitrogen metabolism en-keyword=symbiosis kn-keyword=symbiosis END start-ver=1.4 cd-journal=joma no-vol=72 cd-vols= no-issue=3 article-no= start-page=275 end-page=282 dt-received= dt-revised= dt-accepted= dt-pub-year=2018 dt-pub=201806 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Analysis of All 34 Exons of the SPINK5 Gene in Japanese Atopic Dermatitis Patients en-subtitle= kn-subtitle= en-abstract= kn-abstract= Lympho-epithelial Kazal-type-related inhibitor (LEKTI) is a large multidomain serine protease inhibitor that is expressed in epidermal keratinocytes. Nonsense mutations of the SPINK5 gene, which codes for LEKTI, cause Netherton syndrome, which is characterized by hair abnormality, ichthyosis, and atopy. A single nucleotide polymorphism (SNP) of SPINK5, p.K420E, is reported to be associated with the pathogenesis of atopic dermatitis (AD). We studied all 34 exons of the SPINK5 gene in Japanese 57 AD patients and 50 normal healthy controls. We detected nine nonsynonymous variants, including p.K420E; these variants had already been registered in the SNP database. Among them, p.R654H (n=1) was found as a heterozygous mutation in the AD patients, but not in the control. No new mutation was detected. We next compared the data of the AD patients with data from the Human Genetic Variation Database provided by Kyoto University; a significant difference was found in the frequency of the p.S368N genotype distribution. PolyPhen-2 and SIFT, two algorithms for predicting the functional effects of amino acid substitutions, showed significant scores for p.R654H. Therefore, R654H might be a risk factor for epidermal barrier dysfunction in some Japanese AD patients. en-copyright= kn-copyright= en-aut-name=MorizaneShin en-aut-sei=Morizane en-aut-mei=Shin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OuchidaMamoru en-aut-sei=Ouchida en-aut-mei=Mamoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SunagawaKo en-aut-sei=Sunagawa en-aut-mei=Ko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SugimotoSaeko en-aut-sei=Sugimoto en-aut-mei=Saeko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KobashiMina en-aut-sei=Kobashi en-aut-mei=Mina kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SugiharaSatoru en-aut-sei=Sugihara en-aut-mei=Satoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NomuraHayato en-aut-sei=Nomura en-aut-mei=Hayato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TsujiKazuhide en-aut-sei=Tsuji en-aut-mei=Kazuhide kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=SatoAtsushi en-aut-sei=Sato en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=MiuraYoshihiro en-aut-sei=Miura en-aut-mei=Yoshihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=HattoriHiroaki en-aut-sei=Hattori en-aut-mei=Hiroaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=TadaKotaro en-aut-sei=Tada en-aut-mei=Kotaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=HuhWook-Kang en-aut-sei=Huh en-aut-mei=Wook-Kang kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=SenoAkemi en-aut-sei=Seno en-aut-mei=Akemi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=IwatsukiKeiji en-aut-sei=Iwatsuki en-aut-mei=Keiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= affil-num=1 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil=Department of Molecular Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Nishigawara Dermatology Clinic kn-affil= affil-num=9 en-affil=Sato Dermatology Clinic kn-affil= affil-num=10 en-affil=Miura Dermatology Clinic kn-affil= affil-num=11 en-affil=Hattori Dermatology and Allergy Clinic kn-affil= affil-num=12 en-affil=Tada Dermatology Clinic kn-affil= affil-num=13 en-affil=Dr. Huh's Dermatology Clinic kn-affil= affil-num=14 en-affil=Department of Dermatology, Mitoyo General Hospital kn-affil= affil-num=15 en-affil=Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= en-keyword=atopic dermatitis kn-keyword=atopic dermatitis en-keyword=SPINK5 kn-keyword=SPINK5 en-keyword=LEKTI kn-keyword=LEKTI en-keyword=serine protease inhibitor kn-keyword=serine protease inhibitor en-keyword=epidermal barrier dysfunction kn-keyword=epidermal barrier dysfunction END start-ver=1.4 cd-journal=joma no-vol=9 cd-vols= no-issue=12 article-no= start-page=371 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2017 dt-pub=20171204 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Characterization of a Novel Bat Adenovirus Isolated from Straw-Colored Fruit Bat (Eidolon helvum). en-subtitle= kn-subtitle= en-abstract= kn-abstract= Bats are important reservoirs for emerging zoonotic viruses. For extensive surveys of potential pathogens in straw-colored fruit bats (Eidolon helvum) in Zambia, a total of 107 spleen samples of E. helvum in 2006 were inoculated onto Vero E6 cells. The cell culture inoculated with one of the samples (ZFB06-106) exhibited remarkable cytopathic changes. Based on the ultrastructural property in negative staining and cross-reactivity in immunofluorescence assays, the virus was suspected to be an adenovirus, and tentatively named E. helvum adenovirus 06-106 (EhAdV 06-106). Analysis of the full-length genome of 30,134 bp, determined by next-generation sequencing, showed the presence of 28 open reading frames. Phylogenetic analyses confirmed that EhAdV 06-106 represented a novel bat adenovirus species in the genus Mastadenovirus. The virus shared similar characteristics of low G + C contents with recently isolated members of species Bat mastadenoviruses E, F and G, from which EhAdV 06-106 diverged by more than 15% based on the distance matrix analysis of DNA polymerase amino acid sequences. According to the taxonomic criteria, we propose the tentative new species name "Bat mastadenovirus H". Because EhAdV 06-106 exhibited a wide in vitro cell tropism, the virus might have a potential risk as an emerging virus through cross-species transmission. en-copyright= kn-copyright= en-aut-name=OgawaHirohito en-aut-sei=Ogawa en-aut-mei=Hirohito kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KajiharaMasahiro en-aut-sei=Kajihara en-aut-mei=Masahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NaoNaganori en-aut-sei=Nao en-aut-mei=Naganori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ShigenoAsako en-aut-sei=Shigeno en-aut-mei=Asako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=FujikuraDaisuke en-aut-sei=Fujikura en-aut-mei=Daisuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HangfombeBernard M. en-aut-sei=Hangfombe en-aut-mei=Bernard M. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MweeneAaron S. en-aut-sei=Mweene en-aut-mei=Aaron S. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MutemwaAlisheke en-aut-sei=Mutemwa en-aut-mei=Alisheke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=SquarreDavid en-aut-sei=Squarre en-aut-mei=David kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=YamadaMasao en-aut-sei=Yamada en-aut-mei=Masao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=HigashiHideaki en-aut-sei=Higashi en-aut-mei=Hideaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=SawaHirofumi en-aut-sei=Sawa en-aut-mei=Hirofumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=TakadaAyato en-aut-sei=Takada en-aut-mei=Ayato kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= affil-num=1 en-affil=Department of Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=2 en-affil= Division of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=3 en-affil= Division of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=4 en-affil= Division of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=5 en-affil=Division of Infection and Immunity, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=6 en-affil=Department of Paraclinical Studies, School of Veterinary Medicine, University of Zambia kn-affil= affil-num=7 en-affil=Department of Disease Control, School of Veterinary Medicine, University of Zambia kn-affil= affil-num=8 en-affil= Provincial Veterinary Office, Department of Veterinary Services, Ministry of Fisheries and Livestock kn-affil= affil-num=9 en-affil=Department of National Parks and Wildlife, Ministry of Tourism and Arts kn-affil= affil-num=10 en-affil=Department of Virology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=11 en-affil=Division of Infection and Immunity, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=12 en-affil= Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University kn-affil= affil-num=13 en-affil=Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University kn-affil= en-keyword=Eidolon helvum kn-keyword=Eidolon helvum en-keyword=Zambia kn-keyword=Zambia en-keyword=adenovirus kn-keyword=adenovirus en-keyword=bat kn-keyword=bat END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue= article-no= start-page=12138 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=201607 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A heavy metal P-type ATPase OsHMA4 prevents copper accumulation in rice grain en-subtitle= kn-subtitle= en-abstract= kn-abstract= Rice is a major source of calories and mineral nutrients for over half the world's human population. However, little is known in rice about the genetic basis of variation in accumulation of copper (Cu), an essential but potentially toxic nutrient. Here we identify OsHMA4 as the likely causal gene of a quantitative trait locus controlling Cu accumulation in rice grain. We provide evidence that OsHMA4 functions to sequester Cu into root vacuoles, limiting Cu accumulation in the grain. The difference in grain Cu accumulation is most likely attributed to a single amino acid substitution that leads to different OsHMA4 transport activity. The allele associated with low grain Cu was found in 67 of the 1,367 rice accessions investigated. Identification of natural allelic variation in OsHMA4 may facilitate the development of rice varieties with grain Cu concentrations tuned to both the concentration of Cu in the soil and dietary needs. en-copyright= kn-copyright= en-aut-name=HuangXin-Yuan en-aut-sei=Huang en-aut-mei=Xin-Yuan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=DengFenglin en-aut-sei=Deng en-aut-mei=Fenglin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YamajiNaoki en-aut-sei=Yamaji en-aut-mei=Naoki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=PinsonShannon R.M. en-aut-sei=Pinson en-aut-mei=Shannon R.M. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=Fujii-KashinoMiho en-aut-sei=Fujii-Kashino en-aut-mei=Miho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=DankuJohn en-aut-sei=Danku en-aut-mei=John kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=DouglasAlex en-aut-sei=Douglas en-aut-mei=Alex kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=GuerinotMary Lou en-aut-sei=Guerinot en-aut-mei=Mary Lou kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=SaltDavid E. en-aut-sei=Salt en-aut-mei=David E. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=MaJian Feng en-aut-sei=Ma en-aut-mei=Jian Feng kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil=Institute of Biological and Environmental Sciences, University of Aberdeen kn-affil= affil-num=2 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=3 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=4 en-affil= USDA-ARS Dale Bumpers National Rice Research Center kn-affil= affil-num=5 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= affil-num=6 en-affil=Institute of Biological and Environmental Sciences, University of Aberdeen kn-affil= affil-num=7 en-affil= kn-affil= affil-num=8 en-affil=Department of Biological Sciences, Dartmouth College kn-affil= affil-num=9 en-affil=Institute of Biological and Environmental Sciences, University of Aberdeen kn-affil= affil-num=10 en-affil=Institute of Plant Science and Resources, Okayama University kn-affil= en-keyword=Genetic variation kn-keyword=Genetic variation en-keyword=Natural variation in plants kn-keyword=Natural variation in plants en-keyword=Quantitative trait kn-keyword=Quantitative trait en-keyword=Rice kn-keyword=Rice END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4 article-no= start-page=325 end-page=332 dt-received= dt-revised= dt-accepted= dt-pub-year=2017 dt-pub=201708 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Further Molecular Analysis of G6PD Deficiency Variants in Southern Vietnam and a Novel Variant Designated as G6PD Ho Chi Minh (173 A>G; 58 Asp>Gly): Frequency Distributions of Variants Compared with Those in Other Southeast Asian Countries en-subtitle= kn-subtitle= en-abstract= kn-abstract= We conducted a survey of glucose-6-phosphate dehydrogenase (G6PD) deficiency among newborn babies at Tu Du Hospital, Ho Chi Minh, southern Vietnam. A total of 90 deficient babies were detected, including 85 in the Kinh ethnic group, 4 Chinese, and 1 in the KfHo minority group. In the Kinh ethnic group, G6PD variants such as G6PD Viangchan (n=32), Kaiping (n=11), Canton (n=8), Chinese-5 (n=7), Union (n=5) and Quing Yuan (n=4) were detected. A variant with silent mutations at 1311 C>T and IVS11 nt 93 T>C was also detected in 17 cases. A novel mutation (173 A>G) in exon 4 with a predicted amino acid change of 58 Asp>Gly was also found in a Kinh newborn girl and her father, and it was designated as G6PD Ho Chi Minh. These findings demonstrated that the Kinh ethnic group in southern Vietnam has 8 different G6PD variants, indicating that the members of this group have many ancestors in terms of G6PD variants from Southeast Asia, China, and Oceania. We compared the frequency distribution of G6PD variants in the Kinh population with those of other Southeast Asian populations, and the Kinh populationfs distribution was quite similar to that in the Thai population, but differed from it by the absence of G6PD Mahidol. en-copyright= kn-copyright= en-aut-name=KawamotoFumihiko en-aut-sei=Kawamoto en-aut-mei=Fumihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsuokaHiroyuki en-aut-sei=Matsuoka en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=PhamdNghiem Minh en-aut-sei=Phamd en-aut-mei=Nghiem Minh kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HayashiTaeko en-aut-sei=Hayashi en-aut-mei=Taeko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KasaharaYuichi en-aut-sei=Kasahara en-aut-mei=Yuichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=DungNguyen The en-aut-sei=Dung en-aut-mei=Nguyen The kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KidoYasutoshi en-aut-sei=Kido en-aut-mei=Yasutoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KanbeToshio en-aut-sei=Kanbe en-aut-mei=Toshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=TantularaIndah S. en-aut-sei=Tantulara en-aut-mei=Indah S. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Institute of Tropical Disease, Airlangga University Campus C kn-affil= affil-num=2 en-affil=Division of Medical Zoology, Jichi Medical University kn-affil= affil-num=3 en-affil=Tu Du Hospital kn-affil= affil-num=4 en-affil=Division of Medical Zoology, Jichi Medical University kn-affil= affil-num=5 en-affil=Division of Medical Zoology, Jichi Medical University kn-affil= affil-num=6 en-affil=Vietnam National University School of Medicine kn-affil= affil-num=7 en-affil=Department of Environmental & Preventive Medicine, Oita University Faculty of Medicine kn-affil= affil-num=8 en-affil=Division of Omics Analysis, Nagoya University Graduate School of Medicine kn-affil= affil-num=9 en-affil=Department of Parasitology, Airlangga University Faculty of Medicine kn-affil= en-keyword=G6PD deficiency kn-keyword=G6PD deficiency en-keyword=G6PD variant kn-keyword=G6PD variant en-keyword=southern Vietnam kn-keyword=southern Vietnam en-keyword=Kinh population kn-keyword=Kinh population en-keyword=Southeast Asia kn-keyword=Southeast Asia END start-ver=1.4 cd-journal=joma no-vol=106 cd-vols= no-issue= article-no= start-page=43 end-page=49 dt-received= dt-revised= dt-accepted= dt-pub-year=2017 dt-pub=20170201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Fruit quality control using water management and simple water status detection methods in fruit tree leaves kn-title=‰ΚŽχ‚Ι‚¨‚―‚鐅•ͺƒXƒgƒŒƒX‚Ι‚ζ‚ι ‰ΚŽΐ¬•ͺ‚̐§Œδ‚Ζ—t“ΰ…•ͺ‚ΜŒv‘ͺ en-subtitle= kn-subtitle= en-abstract= kn-abstract=@In fruit production, water status during the maturation season has a significant effect on fruit quality, influencing not only sugar but also organic acid and amino acid contents. Water management technology, therefore, is important for improving fruit quality and maintaining yield in Japan. This study firstly discusses extensive research into the effect of irrigation control on each component of grape fruit juice. Secondly, given that real time monitoring of leaf moisture content is essential to controlling water status, a simple estimation method is described. This method involved fixing a gwater stress indicatorh to the abaxial side of a leaf; the indicator changed color depending on the water status, which could then be evaluated. The water status was assessed against an indicator color scale, based on a property of cobalt iIIj chloride that causes it to change color, from blue to light pink, through a hydration reaction between the indicator sheet and the water evaporating from the leaf to which the indicator sheet is affixed. By using this method, estimates were made of decreases under water stress conditions in the water evaporation rate from satsuma mandarin, grapes, peaches, and Japanese pear, based on the time required for the indicator sheet to change color. Thirdly, a new electrical sensor method to investigate water status in fruit tree leaves was developed, and used to measure electro pulse period; the relationship with transpiration rate was then evaluated using a leaf porometer. Pulse period was found to be consistently correlated with transpiration rate. The results indicate that the water status of fruit tree leaves can be estimated by measuring pulse period. This provides an accurate and quick method for detecting water stress, which could potentially be used for other crops that are particularly sensitive to water stress. en-copyright= kn-copyright= en-aut-name=MorinagaKunihisa en-aut-sei=Morinaga en-aut-mei=Kunihisa kn-aut-name=X‰i–M‹v kn-aut-sei=X‰i kn-aut-mei=–M‹v aut-affil-num=1 ORCID= affil-num=1 en-affil=Graduate School of Environmental and Life Science, Okayama University kn-affil=‰ͺŽR‘εŠw‘εŠw‰@ŠΒ‹«Ά–½‰ΘŠwŒ€‹†‰Θ en-keyword=fruit quality kn-keyword=fruit quality en-keyword=water status kn-keyword=water status en-keyword=patch test kn-keyword=patch test en-keyword=pulse period kn-keyword=pulse period END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=20160325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=CCN2‚Μ“ξœΧ–EƒAƒ~ƒmŽ_‘γŽΣ‚Ι‚¨‚―‚ι–πŠ„ kn-title=Role of CCN2 in Amino Acid Metabolism of Chondrocytes en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=MuraseYurika en-aut-sei=Murase en-aut-mei=Yurika kn-aut-name=‘ΊϋP—F—’ kn-aut-sei=‘ΊϋP kn-aut-mei=—F—’ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue=11 article-no= start-page=e50480 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2013 dt-pub=20130322 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A Catalytic Role of XoxF1 as La3+-Dependent Methanol Dehydrogenase in Methylobacterium extorquens Strain AM1 en-subtitle= kn-subtitle= en-abstract= kn-abstract=In the methylotrophic bacterium Methylobacterium extorquens strain AM1, MxaF, a Ca2+-dependent methanol dehydrogenase (MDH), is the main enzyme catalyzing methanol oxidation during growth on methanol. The genome of strain AM1 contains another MDH gene homologue, xoxF1, whose function in methanol metabolism has remained unclear. In this work, we show that XoxF1 also functions as an MDH and is La3+-dependent. Despite the absence of Ca2+ in the medium strain AM1 was able to grow on methanol in the presence of La3+. Addition of La3+ increased MDH activity but the addition had no effect on mxaF or xoxF1 expression level. We purified MDH from strain AM1 grown on methanol in the presence of La3+, and its N-terminal amino acid sequence corresponded to that of XoxF1. The enzyme contained La3+ as a cofactor. The ƒ’mxaF mutant strain could not grow on methanol in the presence of Ca2+, but was able to grow after supplementation with La3+. Taken together, these results show that XoxF1 participates in methanol metabolism as a La3+-dependent MDH in strain AM1. en-copyright= kn-copyright= en-aut-name=NakagawaTomoyuki en-aut-sei=Nakagawa en-aut-mei=Tomoyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MitsuiRyoji en-aut-sei=Mitsui en-aut-mei=Ryoji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=TaniAkio en-aut-sei=Tani en-aut-mei=Akio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SasaKentaro en-aut-sei=Sasa en-aut-mei=Kentaro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TashiroShinya en-aut-sei=Tashiro en-aut-mei=Shinya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=IwamaTomonori en-aut-sei=Iwama en-aut-mei=Tomonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=HayakawaTakashi en-aut-sei=Hayakawa en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KawaiKeiichi en-aut-sei=Kawai en-aut-mei=Keiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Faculty of Applied Biological Science, Gifu University affil-num=2 en-affil= kn-affil=Department of Biochemistry, Faculty of Science, Okayama University of Science affil-num=3 en-affil= kn-affil=Institute of Plant Science and Resources, Okayama University affil-num=4 en-affil= kn-affil=Faculty of Applied Biological Science, Gifu University affil-num=5 en-affil= kn-affil=Faculty of Applied Biological Science, Gifu University affil-num=6 en-affil= kn-affil=Faculty of Applied Biological Science, Gifu University affil-num=7 en-affil= kn-affil=Faculty of Applied Biological Science, Gifu University affil-num=8 en-affil= kn-affil=Tokai Gakuin University END start-ver=1.4 cd-journal=joma no-vol=105 cd-vols= no-issue= article-no= start-page=21 end-page=27 dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=20160201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Functional analysis of plant immune regulator OsPti1a in rice kn-title=ƒCƒl‘Ο•a«§ŒδˆφŽqOsPti1a ‚Μ §Œδ‹@\‚Μ‰π–Ύ en-subtitle= kn-subtitle= en-abstract= kn-abstract=@An understanding of plant immune systems is important for crop breeding with enhanced disease resistance against pathogen infection. Previous studies reveal that plant has evolved two types of defense mechanisms, which are called gbasal resistanceh and gR?gene mediated resistanceh, for protecting thewrselves from pathogen attack. Recent studies suggest that both defense systems use a common pathway to activate defense responses, however, the downstream components in both pathways are still obscure. @OsPto-interacting protein 1a (OsPti1a), which is a functional ortholog of tomato Pti1, negatively regulates both basal resistance and R?gene mediated resistance in rice. ospti1a mutant shows lesion formation and accompanying defense responses without pathogen infection. OsPti1a is phosphorylated by upstream kinase oxidative signal inducible1 (OsOxi1) and the phosphorylation of OsPti1a has an important role in activating basal resistance against pathogen infection. Additionally, OsOxi1 is phosphorylated by upstream kinase 3?phosphoinotiside-dependent protein kinase 1 (OsPdk1). OsPdk1 has an important role for activating basal resistance against compatible pathogen infection. Therefore, OsPdk1-OsOxi1-OsPti1a phosphorylation cascade regulates proper activation of basal resistance in rice. @Interestingly, OsPti1a localizes at plasma membrane, and cellular localization of OsPti1a has an important function in suppvessing lesion formation. Especially, N?terminal amino acid sequences of OsPti1a have a post-translational modification for binding to plasma membrane. Further, OsPti1a forms complexes with potentially plant immune related proteins at plasma membrane, suggesting that plasma membrane localized OsPti1a probably regulates plant immune complex through its phosphorylation during pathogen infection. en-copyright= kn-copyright= en-aut-name=MatsuiHidenori en-aut-sei=Matsui en-aut-mei=Hidenori kn-aut-name=Όˆδ‰pχ kn-aut-sei=Όˆδ kn-aut-mei=‰pχ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw”_Šw•” en-keyword=lesion mimic mutant kn-keyword=lesion mimic mutant en-keyword=plant immunity kn-keyword=plant immunity en-keyword=phosphorylation kn-keyword=phosphorylation en-keyword=signaling kn-keyword=signaling END start-ver=1.4 cd-journal=joma no-vol=104 cd-vols= no-issue= article-no= start-page=1 end-page=4 dt-received= dt-revised= dt-accepted= dt-pub-year=2015 dt-pub=20150201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Improvement of dye-mediated dehydrogenase activity of pyranose oxidase by site-directed mutagenesis kn-title=•”ˆΚ“ΑˆΩ“I•ΟˆΩ‚Ι‚ζ‚ιƒsƒ‰ƒm[ƒXŽ_‰»y‘f‚̐F‘fˆΛ‘Ά«’E…‘fy‘fŠˆ«‚ΜŒόγ en-subtitle= kn-subtitle= en-abstract= kn-abstract=@Pyranose oxidase (EC 1.1.3.10 ; PROD) catalyzes the oxidation of aldopyranoses at the position C?2 to yield the corresponding 2?keto-aldoses and H2O2 , using oxygen as an electron acceptor. The enzyme shows broad substrate specificity as well as reactivity for 1,5?anhydro?d?glucitol (1,5?AG), which is known as a clinical glycemic marker. It is considered that the reactivity of PROD for 1,5?AG is useful in the development of an amperometric-type biosensor, which is a convenient diagnostic device for selfmonitoring blood glucose (SMBG). However, the levels of dissolved oxygen in blood affect biosensor systems that are equipped with an artificial electron mediator. In the present study, we attempted to develop an O2?insensitive oxidase that would improve the dye-mediated dehydrogenase activity. We performed site-directed mutagenesis on PROD isolated from basidiomycetous fungus No. 52, which generated 11 mutants. The amino acid substitution Q421A exhibited a significant decrease (8.8% of wild type) in its oxidase activity, whereas it maintained its dehydrogenase activity (67% of wild type). In this study, we characterized PROD mutants from basidiomycetous fungus No. 52, which showed improved dye-mediated dehydrogenase activity. en-copyright= kn-copyright= en-aut-name=ArakiToshio en-aut-sei=Araki en-aut-mei=Toshio kn-aut-name=r–؏r—Y kn-aut-sei=r–Ψ kn-aut-mei=r—Y aut-affil-num=1 ORCID= en-aut-name=NakatsukaTomoko en-aut-sei=Nakatsuka en-aut-mei=Tomoko kn-aut-name=’†•Ώ•όŽq kn-aut-sei=’†•Ώ kn-aut-mei=•όŽq aut-affil-num=2 ORCID= en-aut-name=TamuraTakashi en-aut-sei=Tamura en-aut-mei=Takashi kn-aut-name=“c‘Ί—² kn-aut-sei=“c‘Ί kn-aut-mei=—² aut-affil-num=3 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name=ˆξŠ_Œ«“ρ kn-aut-sei=ˆξŠ_ kn-aut-mei=Œ«“ρ aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=’r“cHŒ€Š”Ž‰οŽΠ affil-num=2 en-affil= kn-affil=’r“cHŒ€Š”Ž‰οŽΠ affil-num=3 en-affil= kn-affil=‰ͺŽR‘εŠw”_Šw•” affil-num=4 en-affil= kn-affil=‰ͺŽR‘εŠw”_Šw•” en-keyword=pyranose oxidase kn-keyword=pyranose oxidase en-keyword=1,5-anhydro-d-glucitol kn-keyword=1,5-anhydro-d-glucitol en-keyword=biosensor kn-keyword=biosensor en-keyword=site-directed mutagenesis kn-keyword=site-directed mutagenesis en-keyword=SMBG kn-keyword=SMBG END start-ver=1.4 cd-journal=joma no-vol=61 cd-vols= no-issue=130 article-no= start-page=349 end-page=353 dt-received= dt-revised= dt-accepted= dt-pub-year=2014 dt-pub=201403 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Management Of Peritoneal Effusion by Sealing with a Self-Assembling Nanofiber Polypeptide Following Pelvic Surgery en-subtitle= kn-subtitle= en-abstract= kn-abstract=Background/Aims: PuraMatrix is a synthetic material consisting of 16-amino acid peptides that self-assemble into nanofibers, previously used as a scaffold for functional cell cultures. We conducted a clinical study to determine the safety and sealing properties of PuraMatrix in post-operative lymphorrhea following pelvic surgery in humans. Methodology: A total of 20 patients who underwent rectal cancer resection were analyzed. The study group (n = 10) consisted of patients who received PuraMatrix, matched with a control group (n = 10) of patients operated on conventionally. Results: During the 2 to 3 month follow-up period, there were no abnormal findings or adverse events in any the patients who received PuraMatrix. We found that the patients who received PuraMatrix had significantly reduced post-operative drainage volumes compared with the patients in the control group. Conclusions: PuraMatrix is a safe and effective bio-compatible sealing material for the management of post-operative peritoneal effusion following pelvic surgery. en-copyright= kn-copyright= en-aut-name=KondoYoshitaka en-aut-sei=Kondo en-aut-mei=Yoshitaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NagasakaTakeshi en-aut-sei=Nagasaka en-aut-mei=Takeshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KobayashiSatoru en-aut-sei=Kobayashi en-aut-mei=Satoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KobayashiNaoya en-aut-sei=Kobayashi en-aut-mei=Naoya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=FujiwaraToshiyoshi en-aut-sei=Fujiwara en-aut-mei=Toshiyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Okayama Univ, Dept Surg Gastroenterol, Grad Sch Med Dent & Pharmaceut Sci affil-num=2 en-affil= kn-affil=Okayama Univ, Dept Surg Gastroenterol, Grad Sch Med Dent & Pharmaceut Sci affil-num=3 en-affil= kn-affil=3D Matrix Ltd affil-num=4 en-affil= kn-affil=Okayama Univ, Dept Surg Gastroenterol, Grad Sch Med Dent & Pharmaceut Sci affil-num=5 en-affil= kn-affil=Okayama Univ, Dept Surg Gastroenterol, Grad Sch Med Dent & Pharmaceut Sci en-keyword=Peritoneal effusion kn-keyword=Peritoneal effusion en-keyword=Nanofiber polypeptide kn-keyword=Nanofiber polypeptide en-keyword=Lymphorrhea kn-keyword=Lymphorrhea en-keyword=Pelvic surgery kn-keyword=Pelvic surgery END start-ver=1.4 cd-journal=joma no-vol=126 cd-vols= no-issue=3 article-no= start-page=203 end-page=208 dt-received= dt-revised= dt-accepted= dt-pub-year=2014 dt-pub=20141201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Modulation of neuronal function and neuroprotection by astrocytes kn-title=ƒAƒXƒgƒƒTƒCƒg‚Ι‚ζ‚ι_Œo‹@”\Cό‚Ζƒp[ƒLƒ“ƒ\ƒ“•a‚ł̐_Œo•ΫŒμ en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=AsanumaMasato en-aut-sei=Asanuma en-aut-mei=Masato kn-aut-name=σΐŠ²l kn-aut-sei=σΐ kn-aut-mei=Š²l aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw‘εŠw‰@ˆγŽ•–ςŠw‘‡Œ€‹†‰Θ@_ŒoƒQƒmƒ€Šw en-keyword=ƒAƒXƒgƒƒTƒCƒg kn-keyword=ƒAƒXƒgƒƒTƒCƒg en-keyword=RŽ_‰»–hŒδ‹@\ kn-keyword=RŽ_‰»–hŒδ‹@\ en-keyword=ƒp[ƒLƒ“ƒ\ƒ“•a kn-keyword=ƒp[ƒLƒ“ƒ\ƒ“•a en-keyword=ƒƒ^ƒƒ`ƒIƒlƒCƒ“ kn-keyword=ƒƒ^ƒƒ`ƒIƒlƒCƒ“ en-keyword=Nrf2 kn-keyword=Nrf2 END start-ver=1.4 cd-journal=joma no-vol=68 cd-vols= no-issue=2 article-no= start-page=63 end-page=78 dt-received= dt-revised= dt-accepted= dt-pub-year=2014 dt-pub=201404 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Molecular Simulation Analysis of the Structure Complex of C2 Domains of DKK Family Members and ƒΐ-propeller Domains of LRP5/6:Explaining Why DKK3 Does Not Bind to LRP5/6 en-subtitle= kn-subtitle= en-abstract= kn-abstract=Dickkopf (DKK) proteins interact with low-density lipoprotein receptor-related protein 5/6 (LRP5/6) to modulate WNT signaling. The interaction is mediated by a cysteine-rich domain (C2) in the DKK protein and ƒΐ-propeller domains (PD) of LRP5/6. However, the third member of the DKK family (DKK3) does not bind to LRP5/6. To determine why DKK3 does not bind to the receptor domains, we performed a molecular modeling simulation study including homology modeling, protein-protein docking and molecular dynamics (MD). The computed affinities (ƒ’Gbinding) between the C2 and PD models were consistent with the previously reported experimental results. The C2 model of DKK3 showed the lowest affinity for PD models. Multiple sequence alignment of C2 domains revealed that the DKK3 genes have a unique 7-amino-acid insertion (L249-E255 in human DKK3) and P258 in a finger loop 1 (FL1). Interestingly, the insertion sequence is evolutionally conserved. MD simulations of high-affinity complex models of C2 and PD showed that FL1 directly interacts with the PD models and stabilizes the complex models. We also built a 7-amino-acid-deletion/P258G mutant model of DKK3C2 and estimated its affinities for the PD models. The affinity for human LRP5PD2 was increased by the substitution (ƒ’Gbinding|48.9kcal/mol) and the affinity was compatible with that of high-affinity ligands. The results suggested that the lack of affinity between human DKK3 and human LRP5/6 results from: i) insertion of the 7 amino acids, and ii) P258 in human DKK3. The sequence differences thus suggest an explanation for this unique property of DKK3. en-copyright= kn-copyright= en-aut-name=FujiiYasuyuki en-aut-sei=Fujii en-aut-mei=Yasuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HoshinoTyuji en-aut-sei=Hoshino en-aut-mei=Tyuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KumonHiromi en-aut-sei=Kumon en-aut-mei=Hiromi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil= kn-affil=Innovation Center Okayama for Nanobio-Targeted Therapy, Okayama University Graduate School of Medicine affil-num=2 en-affil= kn-affil=Department of Physical Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University affil-num=3 en-affil= kn-affil=Innovation Center Okayama for Nanobio-Targeted Therapy, Okayama University Graduate School of Medicine en-keyword=DKK3 kn-keyword=DKK3 en-keyword=molecular modeling kn-keyword=molecular modeling en-keyword=protein-protein docking kn-keyword=protein-protein docking en-keyword=LRP5/6 kn-keyword=LRP5/6 END start-ver=1.4 cd-journal=joma no-vol=49 cd-vols= no-issue=1 article-no= start-page=1 end-page=8 dt-received= dt-revised= dt-accepted= dt-pub-year=2012 dt-pub=201201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Ca2+-independent syntaxin binding to the C2B effector region of synaptotagmin en-subtitle= kn-subtitle= en-abstract= kn-abstract=Although synaptotagmin I, which is a calcium (Ca2+)-binding synaptic vesicle protein, may trigger soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-mediated synaptic vesicle exocytosis, the mechanisms underlying the interaction between these proteins remain controversial, especially with respect to the identity of the protein(s) in the SNARE complex that bind(s) to synaptotagmin and whether Ca2+ is required for their highly effective binding. To address these questions, native proteins were solubilized, immunoprecipitated from rat brain extracts, and analyzed by immunoblotting. SNARE complexes comprising syntaxin 1, 25-kDa synaptosomal-associated protein (SNAP-25), and synaptobrevin 2 were coprecipitzted with synaptotagmin I in the presence of ethylene glycol tetraacetic acid. The amount of cop recipitated proteins was significantly unaltered by the addition of Ca2+ to the brain extract. To identify the component of the SNARE complex that bound to synaptotagmin, SNARE was coexpressed with synaptotagmin in HEK293 cells and immunoprecipitated. Syntaxin, but not SNAP-25 and synaptobrevin, bound to synaptotagmin in a Ca2+-independent manner, and the binding was abolished in the presence of 1 M NaCl. Synaptotagmin contains 2 Ca2+-binding domains (C(2)A, C2B). Mutating the positively charged lysine residues in the putative effector-binding region of the C2B domain, which are critical for transmitter release, markedly inhibited synaptotagmin-syntaxin binding, while similar mutations in the C(2)A domain had no effect on binding. Synaptotagmin-syntaxin binding was reduced by mutating multiple negatively charged glutamate residues in the amino-terminal half of the syntaxin SNARE motif. These results indicate that synaptotagmin I binds to syntaxin 1 electrostatically through its C2B domain effector region in a Ca2+-independent fashion, providing biochemical evidence that synaptotagmin I binds SNARE complexes before Ca2+ influx into presynaptic nerve terminals. en-copyright= kn-copyright= en-aut-name=MasumotoToshio en-aut-sei=Masumoto en-aut-mei=Toshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SuzukiKoichiro en-aut-sei=Suzuki en-aut-mei=Koichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OhmoriIori en-aut-sei=Ohmori en-aut-mei=Iori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MichiueHiroyuki en-aut-sei=Michiue en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TomizawaKazuhito en-aut-sei=Tomizawa en-aut-mei=Kazuhito kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=FujimuraAtsushi en-aut-sei=Fujimura en-aut-mei=Atsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=NishikiTei-ichi en-aut-sei=Nishiki en-aut-mei=Tei-ichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MatsuiHideki en-aut-sei=Matsui en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=2 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=3 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=4 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=5 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=6 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=7 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol affil-num=8 en-affil= kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Physiol en-keyword=Neurotransmitter release kn-keyword=Neurotransmitter release en-keyword=Synaptic vesicle kn-keyword=Synaptic vesicle en-keyword=Exocytosis kn-keyword=Exocytosis en-keyword=SNAP-25 kn-keyword=SNAP-25 en-keyword=Synaptobrevin kn-keyword=Synaptobrevin END start-ver=1.4 cd-journal=joma no-vol=16 cd-vols= no-issue=2 article-no= start-page=81 end-page=89 dt-received= dt-revised= dt-accepted= dt-pub-year=2009 dt-pub=200904 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Development of 5006 Full-Length CDNAs in Barley: A Tool for Accessing Cereal Genomics Resources en-subtitle= kn-subtitle= en-abstract= kn-abstract=A collection of 5006 full-length (FL) cDNA sequences was developed in barley. Fifteen mRNA samples from various organs and treatments were pooled to develop a cDNA library using the CAP trapper method. More than 60% of the clones were confirmed to have complete coding sequences, based on comparison with rice amino acid and UniProt sequences. Blastn homologies (E < 1E-5) to rice genes and Arabidopsis genes were 89 and 47%, respectively. Of the 5028 possible amino acid sequences derived from the 5006 FLcDNAs, 4032 (80.2%) were classified into 1678 GreenPhyl multigenic families. There were 555 cDNAs showing low homology to both rice and Arabidopsis. Gene ontology annotation by InterProScan indicated that many of these cDNAs (71%) have no known molecular functions and may be unique to barley. The cDNAs showed high homology to Barley 1 GeneChip oligo probes (81%) and the wheat gene index (84%). The high homology between FLcDNAs (27%) and mapped barley expressed sequence tag enabled assigning linkage map positions to 151-233 FLcDNAs on each of the seven barley chromosomes. These comprehensive barley FLcDNAs provide strong platform to connect preexisting genomic and genetic resources and accelerate gene identification and genome analysis in barley and related species. en-copyright= kn-copyright= en-aut-name=SatoKazuhiro en-aut-sei=Sato en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=Shin-ITadasu en-aut-sei=Shin-I en-aut-mei=Tadasu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SekiMotoaki en-aut-sei=Seki en-aut-mei=Motoaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ShinozakiKazuo en-aut-sei=Shinozaki en-aut-mei=Kazuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YoshidaHideya en-aut-sei=Yoshida en-aut-mei=Hideya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TakedaKazuyoshi en-aut-sei=Takeda en-aut-mei=Kazuyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=YamazakiYukiko en-aut-sei=Yamazaki en-aut-mei=Yukiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=ConteMatthieu en-aut-sei=Conte en-aut-mei=Matthieu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KoharaYuji en-aut-sei=Kohara en-aut-mei=Yuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil= kn-affil=Okayama Univ, Bioresources Res Inst affil-num=2 en-affil= kn-affil=Natl Inst Genet affil-num=3 en-affil= kn-affil=RIKEN, Plant Sci Ctr affil-num=4 en-affil= kn-affil=RIKEN, Plant Sci Ctr affil-num=5 en-affil= kn-affil=Okayama Univ, Bioresources Res Inst affil-num=6 en-affil= kn-affil=Okayama Univ, Bioresources Res Inst affil-num=7 en-affil= kn-affil=Natl Inst Genet affil-num=8 en-affil= kn-affil=Int Rice Res Inst, Crop Res Informat Lab affil-num=9 en-affil= kn-affil=Natl Inst Genet en-keyword=full-length cDNA kn-keyword=full-length cDNA en-keyword=Hordeum vulgare kn-keyword=Hordeum vulgare en-keyword=mRNA kn-keyword=mRNA en-keyword=gene ontology kn-keyword=gene ontology END start-ver=1.4 cd-journal=joma no-vol=103 cd-vols= no-issue= article-no= start-page=5 end-page=9 dt-received= dt-revised= dt-accepted= dt-pub-year=2014 dt-pub=20140201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Characterization of L-Arginine Oxidase Made from L-Glutamate Oxidase kn-title=‚ŠξŽΏ“ΑˆΩ«L-ƒOƒ‹ƒ^ƒ~ƒ“Ž_ƒIƒLƒVƒ_[ƒ[‚ζ‚θμ¬‚΅‚½ŠξŽΏ“ΑˆΩ«‰ό•Οy‘fL-ƒAƒ‹ƒMƒjƒ“ƒIƒLƒVƒ_[ƒ[‚̐«ŽΏŒŸ“’ en-subtitle= kn-subtitle= en-abstract= kn-abstract=@L?Glutamate oxidase (LGOX) from Streptomyces sp. X?119?6 has strict substrate specificity toward L?glutamate. Recently, we solved the X?ray crystal structure of LGOX and this revealed that Arg305 in the active site is the key residue involved in substrate recognition. Therefore, we created 19 mutant enzymes of R305X?LGOX by saturation mutagenesis. One of them R305D?LGOX, Arg305 substituted with Asp exhibited oxidase activity for L?Arg. Optimum pH of R305D?LGOX mutant enzyme was pH 8.5. Interestingly, the activity of R305D?LGOX toward L?Arg was inhibited by phosphate. And furthermore, the substrate specificity of R305D?LGOX was affected by using buffer. The results of inhibition analysis suggest, that phosphate is a competitive inhibitor of R305D?LGOX when L?Arg is used as substrate. Kinetic analysis of R305D?LGOX showed that Km value and kcat value of R305D?LGOX toward l-Arg were 0.68 mM and 6.7 s-1 respectively. In this study, we showed that R305D?LGOX mutant enzyme is a novel l-arginine oxidase and useful for l-arginine biosensor. en-copyright= kn-copyright= en-aut-name=NakaiRyuichiro en-aut-sei=Nakai en-aut-mei=Ryuichiro kn-aut-name=’†ˆδ—²ˆκ˜Y kn-aut-sei=’†ˆδ kn-aut-mei=—²ˆκ˜Y aut-affil-num=1 ORCID= en-aut-name=FujinoShihoko en-aut-sei=Fujino en-aut-mei=Shihoko kn-aut-name=“‘–μŽu•ΫŽq kn-aut-sei=“‘–μ kn-aut-mei=Žu•ΫŽq aut-affil-num=2 ORCID= en-aut-name=UtsumiTomohiro en-aut-sei=Utsumi en-aut-mei=Tomohiro kn-aut-name=“ΰŠC—FG kn-aut-sei=“ΰŠC kn-aut-mei=—FG aut-affil-num=3 ORCID= en-aut-name=TamuraTakashi en-aut-sei=Tamura en-aut-mei=Takashi kn-aut-name=“c‘Ί—² kn-aut-sei=“c‘Ί kn-aut-mei=—² aut-affil-num=4 ORCID= en-aut-name=KusakabeaHitoshi en-aut-sei=Kusakabea en-aut-mei=Hitoshi kn-aut-name=“ϊ‰Ί•”‹Ο kn-aut-sei=“ϊ‰Ί•” kn-aut-mei=‹Ο aut-affil-num=5 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name=ˆξŠ_Œ«“ρ kn-aut-sei=ˆξŠ_ kn-aut-mei=Œ«“ρ aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw affil-num=2 en-affil= kn-affil=‰ͺŽR‘εŠw affil-num=3 en-affil= kn-affil=‰ͺŽR‘εŠw affil-num=4 en-affil= kn-affil=‰ͺŽR‘εŠw affil-num=5 en-affil= kn-affil=‡ŠƒGƒ“ƒUƒCƒ€ƒZƒ“ƒT affil-num=6 en-affil= kn-affil=‰ͺŽR‘εŠw en-keyword=L-glutamate oxidase kn-keyword=L-glutamate oxidase en-keyword=L-arginine oxidase kn-keyword=L-arginine oxidase en-keyword=biosensor kn-keyword=biosensor en-keyword=modified substrate specificity kn-keyword=modified substrate specificity en-keyword=L-amino acid oxidase kn-keyword=L-amino acid oxidase END start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=2 article-no= start-page=93 end-page=98 dt-received= dt-revised= dt-accepted= dt-pub-year=2013 dt-pub=201304 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The Genetic Diversity of Helicobacter pylori Virulence Genes Is Not Associated with Gastric Atrophy Progression en-subtitle= kn-subtitle= en-abstract= kn-abstract=Atrophy of the gastric mucosa is a precursor of intestinal-type gastric cancer, and Helicobacter pylori infection causes atrophic gastritis. The aim of this study was to determine whether the genetic diversity of H. pylori virulence genes is associated with the development and progression of gastric atrophy in humans. We isolated and cultured H. pylori strains from patients with gastric ulcer and duodenal ulcer accompanied by atrophic gastritis in background mucosa. H. pylori strains were stored at |80Ž prior to the experiments being carried out. We analyzed iceA, babA, vacA, cagA, and cagE genes by PCR. The cagA gene was analyzed through sequencing of the C-terminal region containing the EPIYA motif, which is related to tyrosine phosphorylation. Severe atrophy was observed in patients with gastric ulcer. The major phenotype of the vacA gene was s1c/m1 (93΅). The cagA gene was detected in all strains. The cagE gene was not detected in 2 and 5 strains from the mild cases and severe cases, respectively. The major cagA EPIYA motif, which is amino acids repeat in the C terminus, was the A-B-D type (44 of 58 strains). The virulence genes were not statistically associated with the severity of atrophy in the background gastric mucosa in humans. Not only identification of bacterial virulence factors but also studies of the host response will be necessary to investigate the progression of gastric atrophy and subsequent cancer development in humans. en-copyright= kn-copyright= en-aut-name=KitaMasahide en-aut-sei=Kita en-aut-mei=Masahide kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 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=2 ORCID= en-aut-name=OkadaHiroyuki en-aut-sei=Okada en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TakeSusumu en-aut-sei=Take en-aut-mei=Susumu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TakenakaRyuta en-aut-sei=Takenaka en-aut-mei=Ryuta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KawaharaYoshiro en-aut-sei=Kawahara en-aut-mei=Yoshiro 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= 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=YamamotoKazuhide en-aut-sei=Yamamoto en-aut-mei=Kazuhide kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil= kn-affil=Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=2 en-affil= kn-affil=Graduate School of Health Sciences, Okayama University affil-num=3 en-affil= kn-affil=Department of Endoscopy, Okayama University Hospital affil-num=4 en-affil= kn-affil=Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=5 en-affil= kn-affil=Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=6 en-affil= kn-affil=cDepartment of Endoscopy, Okayama University Hospital affil-num=7 en-affil= kn-affil=Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 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 Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences en-keyword=Helicobacter pylori kn-keyword=Helicobacter pylori en-keyword=virulence genes kn-keyword=virulence genes en-keyword=chronic atrophic gastritis kn-keyword=chronic atrophic gastritis END 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=2011 cd-vols= no-issue=1 article-no= start-page=5 end-page=11 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20110809 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Detection of in situ cleaved p115 with the cut specific antibodies in rapid protein inactivation system by tobacco etch viral protease cleavage en-subtitle= kn-subtitle= en-abstract= kn-abstract=Gene perturbation methods are commonly used in the study of gene and protein function. The authors of this paper recently developed a rapid protein inactivation technique utilizing tobacco etch virus (TEV)-derived protease. TEV protease recognizes the ENLYFQG (Glu-Asn-Leu-Tyr-Phe-Gln-Gly) amino acid sequence and specifically cleaves between Q and G. The authors developed antibodies that recognize the cleaved TEV (ENLYFQ) sequence, both in vitro and in vivo, but do not bind to uncleaved TEV (ENLYFQG). Using these antibodies, in situ protein cleavage was successfully detected. These antibodies used in combination with the TEV protease may be a useful complement to other perturbation methods. en-copyright= kn-copyright= en-aut-name=KoreishiMayuko en-aut-sei=Koreishi en-aut-mei=Mayuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HonjoYasuko en-aut-sei=Honjo en-aut-mei=Yasuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SatohAyano en-aut-sei=Satoh en-aut-mei=Ayano kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University kn-affil= affil-num=2 en-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University kn-affil= affil-num=3 en-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University kn-affil= en-keyword=TEV protease kn-keyword=TEV protease en-keyword=Golgi kn-keyword=Golgi en-keyword=golgins kn-keyword=golgins en-keyword=microinjection kn-keyword=microinjection en-keyword=recombinant proteins kn-keyword=recombinant proteins END start-ver=1.4 cd-journal=joma no-vol=92 cd-vols= no-issue=2 article-no= start-page=361 end-page=369 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=201102 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Whole-genome characterization of human group C rotaviruses: identification of two lineages in the VP3 gene en-subtitle= kn-subtitle= en-abstract= kn-abstract=Group C rotavirus (GCRV) is distributed worldwide as an enteric pathogen in humans and animals. However, to date, whole-genome sequences are available only for a human strain (Bristol) and a porcine strain (Cowden). To investigate the genetic diversity of human GCRVs, nearly full-length sequences of all 11 RNA segments were determined for human GCRVs detected recently in India (v508), Bangladesh (BS347), China (Wu82 and YNR001) and Japan (OH567 and BK0830) and analysed phylogenetically with sequence data for GCRVs published previously. All the RNA segments of human GCRV strains except for the VP3 gene showed high levels of conservation (>93?% nucleotide sequence identity, >92?% amino acid sequence identity), belonging to a single genetic cluster distinct from those of animal GCRVs. In contrast, the VP3 genes of human GCRVs could be discriminated into two clusters, designated M2 and M3, that were distinguished phylogenetically from those of porcine and bovine GCRVs (clusters M1 and M4, respectively). Between M2 and M3, amino acid sequence identity of the VP3 gene was 84.1?84.7?%, whereas high identities were observed within each cluster (92.3?97.6?% for M2, 98.2?99.3?% for M3). Sequence divergence among the four VP3 clusters was observed throughout the amino acid sequence except for conserved motifs, including those possibly related to enzyme functions of VP3. The presence of obvious genetic diversity only in the VP3 gene among human GCRVs suggested that either the M2 or M3 VP3 gene of human GCRVs might have been derived through reassortment from an animal GCRV or from an unidentified human GCRV strain belonging to a novel genogroup. en-copyright= kn-copyright= en-aut-name=YamamotoDai en-aut-sei=Yamamoto en-aut-mei=Dai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=GhoshSouvik en-aut-sei=Ghosh en-aut-mei=Souvik kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KuzuyaMitsutaka en-aut-sei=Kuzuya en-aut-mei=Mitsutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=WangYuan-Hong en-aut-sei=Wang en-aut-mei=Yuan-Hong kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=ZhouXuan en-aut-sei=Zhou en-aut-mei=Xuan kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=Chawla-SarkarMamta en-aut-sei=Chawla-Sarkar en-aut-mei=Mamta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=PaulShyamal Kumar en-aut-sei=Paul en-aut-mei=Shyamal Kumar kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=IshinoMasaho en-aut-sei=Ishino en-aut-mei=Masaho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KobayashiNobumichi en-aut-sei=Kobayashi en-aut-mei=Nobumichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine affil-num=2 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine affil-num=3 en-affil= kn-affil=Okayama Prefectural Institute for Environmental Science and Public Health affil-num=4 en-affil= kn-affil=Wuhan Centers for Disease Prevention and Control affil-num=5 en-affil= kn-affil=Wuhan Centers for Disease Prevention and Control affil-num=6 en-affil= kn-affil=National Institute of Cholera and Enteric Diseases affil-num=7 en-affil= kn-affil=Mymensingh Medical College affil-num=8 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine affil-num=9 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine END start-ver=1.4 cd-journal=joma no-vol=91 cd-vols= no-issue=7 article-no= start-page=1772 end-page=1781 dt-received= dt-revised= dt-accepted= dt-pub-year=2010 dt-pub=201007 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Analysis of genetic diversity and molecular evolution of human group B rotaviruses based on whole genome segments en-subtitle= kn-subtitle= en-abstract= kn-abstract=Group B rotavirus (GBR) is a rare enteric pathogen that causes severe diarrhoea, primarily in adults. Nearly full-length sequences of all 11 RNA segments were determined for human GBRs detected recently in India (IDH-084 in 2007, IC-008 in 2008), Bangladesh (Bang117 in 2003) and Myanmar (MMR-B1 in 2007), and analysed phylogenetically with the sequence data of GBRs reported previously. All RNA segments of GBR strains from India, Bangladesh and Myanmar showed >95?% nucleotide sequence identities. Among the 11 RNA segments, the VP6 and NSP2 genes showed the highest identities (>98?%), whilst the lowest identities were observed in the NSP4 gene (96.1?%), NSP5 gene (95.6?%) and VP8*-encoding region of the VP4 gene (95.9?%). Divergent or conserved regions in the deduced amino acid sequences of GBR VP1?VP4 and NSP1?NSP5 were similar to those in group A rotaviruses (GARs), and the functionally important motifs and structural characteristics in viral proteins known for GAR were conserved in all of the human GBRs. These findings suggest that, whilst the degree of genetic evolution may be dependent on each RNA segment, human GBR may have been evolving in a similar manner to GAR, associated with the similar functional roles of individual viral proteins. en-copyright= kn-copyright= en-aut-name=YamamotoDai en-aut-sei=Yamamoto en-aut-mei=Dai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=GhoshSouvik en-aut-sei=Ghosh en-aut-mei=Souvik kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=GaneshBalasubramanian en-aut-sei=Ganesh en-aut-mei=Balasubramanian kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KrishnanTriveni en-aut-sei=Krishnan en-aut-mei=Triveni kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=Chawla-SarkarMamta en-aut-sei=Chawla-Sarkar en-aut-mei=Mamta kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=AlamMohammed Mahbub en-aut-sei=Alam en-aut-mei=Mohammed Mahbub kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=AungTin Sabai en-aut-sei=Aung en-aut-mei=Tin Sabai kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KobayashiNobumichi en-aut-sei=Kobayashi en-aut-mei=Nobumichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine affil-num=2 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine affil-num=3 en-affil= kn-affil=National Institute of Cholera and Enteric Diseases affil-num=4 en-affil= kn-affil=National Institute of Cholera and Enteric Diseases affil-num=5 en-affil= kn-affil=National Institute of Cholera and Enteric Diseases affil-num=6 en-affil= kn-affil=Bangladesh Agricultural University affil-num=7 en-affil= kn-affil=National Health Laboratory affil-num=8 en-affil= kn-affil=Department of Hygiene, Sapporo Medical University School of Medicine END start-ver=1.4 cd-journal=joma no-vol=57 cd-vols= no-issue=6 article-no= start-page=904 end-page=908 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=201105 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Inactivation of Vibrio vulnificus hemolysin through mutation of the N- or C-terminus of the lectin-like domain en-subtitle= kn-subtitle= en-abstract= kn-abstract=Vibrio vulnificus is an etiological agent causing serious systemic infections in the immunocompromised humans or cultured eels. This species commonly produces a hemolytic toxin consisting of the cytolysin domain and the lectin-like domain. For hemolysis, the lectin-like domain specifically binds to cholesterol in the erythrocyte membrane, and to form a hollow oligomer, the toxin is subsequently assembled on the membrane. The cytolysin domain is essential for the process to form the oligomer. Three-dimensional structure model revealed that two domains connected linearly and the C-terminus was located near to the joint of the domains. Insertion of amino acid residues between two domains was found to cause inactivation of the toxin. In the C-terminus, deletion, substitution or addition of an amino acid residue also elicited reduction of the activity. However, the cholesterol-binding ability was not affected by the mutations. These results suggest that mutation of the C- or N-terminus of the lectin-like domain may result in blockage of the toxin assembly. en-copyright= kn-copyright= en-aut-name=MiyoshiShin-ichi en-aut-sei=Miyoshi en-aut-mei=Shin-ichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AbeYuki en-aut-sei=Abe en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SenohMitsutoshi en-aut-sei=Senoh en-aut-mei=Mitsutoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MizunoTamaki en-aut-sei=Mizuno en-aut-mei=Tamaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MaeharaYoko en-aut-sei=Maehara en-aut-mei=Yoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=NakaoHiroshi en-aut-sei=Nakao en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=2 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=3 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=4 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=5 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=6 en-affil= kn-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University en-keyword=Vibrio vulnificus kn-keyword=Vibrio vulnificus en-keyword=Hemolysin kn-keyword=Hemolysin en-keyword=Cell-free translation kn-keyword=Cell-free translation en-keyword=Site-directed mutagenesis kn-keyword=Site-directed mutagenesis END start-ver=1.4 cd-journal=joma no-vol=16 cd-vols= no-issue=10 article-no= start-page=8815 end-page=8832 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20111020 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Studies on the Synthesis of DMAP Derivatives by Diastereoselective Ugi Reactions en-subtitle= kn-subtitle= en-abstract= kn-abstract=Diastereoselective Ugi reactions of DMAP-based aldehydes with ƒΏ-amino acids and tert-butyl isocyanide were examined. The reactions of 4-(dimethylamino)-2-pyridine-carboxaldehyde with various ƒΏ-amino acids afforded 2-substituted DMAP derivatives with low diastereoselectivity. On the contrary, reactions with 4-(dimethylamino)-3-pyridine-carboxaldehyde delivered 3-substituted DMAP derivatives with moderate to high diastereoselectivity. The combination of ƒΏ-amino acid and DMAP-based aldehyde is thus important to achieve high diastereoselectivity. Kinetic resolution of a secondary alcohol using a chiral DMAP derivative obtained through these reactions was also examined. en-copyright= kn-copyright= en-aut-name=MandaiHiroki en-aut-sei=Mandai en-aut-mei=Hiroki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IrieShunsuke en-aut-sei=Irie en-aut-mei=Shunsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MitsudoKoichi en-aut-sei=Mitsudo en-aut-mei=Koichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SugaSeiji en-aut-sei=Suga en-aut-mei=Seiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University affil-num=2 en-affil= kn-affil=Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University affil-num=3 en-affil= kn-affil=Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University affil-num=4 en-affil= kn-affil=Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University en-keyword=multicomponent reaction kn-keyword=multicomponent reaction en-keyword=Ugi reaction kn-keyword=Ugi reaction en-keyword=chiral DMAP kn-keyword=chiral DMAP en-keyword=kinetic resolution kn-keyword=kinetic resolution END start-ver=1.4 cd-journal=joma no-vol= cd-vols= no-issue= article-no= start-page= end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20111231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=•ͺŠς½ƒAƒ~ƒmŽ_‚Šά—L‰h—{ά‚͐Ά‘ΜŠΜˆΪApŒγ‘Šϊ‚Ι‚¨‚―‚ι‰h—{αŠQE‘γŽΣˆΩν‚π‰ό‘P‚·‚ι kn-title=Branched-chain amino acid-enriched nutrients improve nutritional and metabolic abnormalities in the early post-transplant period after living donor liver transplantation en-subtitle= kn-subtitle= en-abstract= kn-abstract= en-copyright= kn-copyright= en-aut-name=YoshidaRyuichi en-aut-sei=Yoshida en-aut-mei=Ryuichi kn-aut-name=‹g“c—΄ˆκ kn-aut-sei=‹g“c kn-aut-mei=—΄ˆκ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠw END start-ver=1.4 cd-journal=joma no-vol=101 cd-vols= no-issue= article-no= start-page=1 end-page=6 dt-received= dt-revised= dt-accepted= dt-pub-year=2012 dt-pub=20120201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Purification and Characterization of Thermostable Amidase from Thermus sp.O-3-1 kn-title=D”M«Χ‹ΫThermus sp.O-3-1—R—ˆ‘Ο”M«ƒAƒ~ƒ_[ƒ[‚̐Έ»‹y‚ѐ«ŽΏŒŸ“’ en-subtitle= kn-subtitle= en-abstract=D”M«Χ‹ΫThermus sp.O-3-1 —R—ˆ‚Μ‘Ο”M«ƒAƒ~ƒ_[ƒ[ˆβ“`Žq‚π‘ε’°‹Ϋ’†‚ΙƒNƒ[ƒjƒ“ƒO‚΅C‚»‚Μ‰–Šξ”z—ρ‚πŒˆ’θ‚΅‚½Dami ˆβ“`Žq‚Ν930 bp ‚©‚η‚Θ‚θC310ƒAƒ~ƒmŽ_‚πƒR[ƒh‚΅‚Δ‚’‚½D–{y‘f‚Μ•ͺŽq—Κ‚Ν33C089 Da‚Ε‚ ‚ι‚Ζ—\‘z‚³‚κ‚½DThermus sp.O-3-1 —R—ˆƒAƒ~ƒ_[ƒ[‚π‘ε’°‹Ϋ‚ŐΆŽY‚³‚ΉC”Mˆ—‚ΖDEAE-ƒgƒˆƒp[ƒ‹650M‰AƒCƒIƒ“ŒπŠ·ƒJƒ‰ƒ€“™‚Ι‚ζ‚萸»‚΅‚½DƒQƒ‹ΰh‰ίƒNƒƒ}ƒgƒOƒ‰ƒtƒB[‚ΖSDS-PAGE ‚ΜŒ‹‰Κ‚©‚η–{y‘f‚Ν•ͺŽqŽΏ—Κ33 kDa ‚ΜƒTƒuƒ†ƒjƒbƒg2•ͺŽq‚©‚η‚Θ‚ιƒ_ƒCƒ}[\‘’‚π—L‚΅‚Δ‚’‚ι‚±‚Ζ‚ͺ–Ύ‚η‚©‚Ζ‚Θ‚Α‚½DΈ»y‘f‚Μ”Mˆΐ’萫‚Ν80Ž‚܂ŁCpH ˆΐ’萫‚Ν7.0`10.0‚Ε‚ ‚θCˆΐ’萫‚Μ ‚‚’y‘f‚Ε‚ ‚Α‚½DΕ“K‰·“x‚Ν90ŽCΕ“K pH ‚Ν9.0‚Ε‚  ‚Α‚½DEDTA ‚Ι‚ζ‚θŠˆ«‚ͺ’˜‚΅‚­‘jŠQ‚³‚κCCo(2+)‚βNi(2+)CMn(2+)‚Ι‚ζ‚Α‚ΔŠˆ«‚Μ‰ρ•œCŒόγ‚ͺŒ©‚η‚κ‚½‚½‚߁C–{y‘f‚Ν‹ΰ‘y‘f‚Ε‚ ‚ι‚±‚Ζ‚ͺŽ¦΄‚³‚κ‚½DŠξŽΏ“ΑˆΩ«‚ΜŒŸ“’ ‚ΜŒ‹‰ΚCL-Leu-pNA ‚ζ‚θ‚ΰD-Leu-pNA ‚Ι‘Ξ‚΅‚č‚‚’Šˆ«‚πŽ¦‚΅‚½‚½‚߁C–{y‘f‚ͺ‚c-ƒAƒ~ƒmŽ_ŠξŽΏ‚Ι“ΑˆΩ«‚πŽ‚ΒƒAƒ~ƒ_[ƒ[‚Ε‚ ‚ι‚±‚Ζ‚ͺ”»–Ύ‚΅‚½D–{y‘f‚Ν‘Ο”M«‚π—L‚·‚郆ƒj[ƒN‚Θ‚c-ƒAƒ~ƒmŽ_ƒAƒ~ƒ_[ƒ[‚Ε‚ ‚θC‘ŒγŽY‹Ζ—˜—p‚ͺŠϊ‘‚³‚κ‚ιD kn-abstract=The gene encoding a thermostable amidase (EC 3.5.1.4) from thermophilic bacterium Thermus sp.O-3-1, was cloned and expressed in Escherichia coli JM109. The cloned amidase gene (ami) is 930 bp and encodes a protein composed of 310 amino acids. The protein is predicted to have a molecular mass of 33,089 Da. The amidase from Thermus sp.O-3-1 was purified by heat treatment and DEAE Toyopearl 650M column chromatography. The molecular mass of the native enzyme was estimated to be about 70 kDa by gel filtration chromatography, indicating that the enzyme has a homodimeric structure. The purified enzyme was stable up to 80‹C and within a pH range from 7.0 to 10.0. The optimum temperature and pH for enzyme activity were 90‹C, and 9.0, respectively. The enzyme was strongly inhibited by the metal-chelating compound EDTA. The activity of the EDTA-treated enzyme was reactivated by the addition of Co(2+), Ni(2+) and Mn(2+) ions. Therefore the enzyme was predicted to be metalloenzyme. Finally, as a result of investigation into substrate specificity, the purified enzyme was suggested to be D-amino acid specific amidase, as it showed higher activity toward D-Leu-pNA than L-Leu-pNA. en-copyright= kn-copyright= en-aut-name=KobayashiFumiaki en-aut-sei=Kobayashi en-aut-mei=Fumiaki kn-aut-name=¬—ΡŽj–Ύ kn-aut-sei=¬—Ρ kn-aut-mei=Žj–Ύ aut-affil-num=1 ORCID= en-aut-name=AomineHiroki en-aut-sei=Aomine en-aut-mei=Hiroki kn-aut-name=Β•τO‹N kn-aut-sei=Β•τ kn-aut-mei=O‹N aut-affil-num=2 ORCID= en-aut-name=MizunashiWataru en-aut-sei=Mizunashi en-aut-mei=Wataru kn-aut-name=…–³Β kn-aut-sei=…–³ kn-aut-mei=Β aut-affil-num=3 ORCID= en-aut-name=YuFujio en-aut-sei=Yu en-aut-mei=Fujio kn-aut-name=“’•s“ρ•v kn-aut-sei=“’ kn-aut-mei=•s“ρ•v aut-affil-num=4 ORCID= en-aut-name=TamuraTakashi en-aut-sei=Tamura en-aut-mei=Takashi kn-aut-name=“c‘Ί—² kn-aut-sei=“c‘Ί kn-aut-mei=—² aut-affil-num=5 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name=ˆξŠ_Œ«“ρ kn-aut-sei=ˆξŠ_ kn-aut-mei=Œ«“ρ aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil= affil-num=2 en-affil= kn-affil= affil-num=3 en-affil= kn-affil=(Š”)ŽO•HƒŒƒCƒˆƒ“ affil-num=4 en-affil= kn-affil= affil-num=5 en-affil= kn-affil=‰ͺŽR‘εŠw affil-num=6 en-affil= kn-affil=‰ͺŽR‘εŠw en-keyword=amidase kn-keyword=amidase en-keyword=thermostable enzyme kn-keyword=thermostable enzyme en-keyword=Thermus kn-keyword=Thermus en-keyword=D-amino acid specific amidase kn-keyword=D-amino acid specific amidase END start-ver=1.4 cd-journal=joma no-vol=20 cd-vols= no-issue=11 article-no= start-page=1347 end-page=1354 dt-received= dt-revised= dt-accepted= dt-pub-year=2003 dt-pub=200311 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Phase Shifts of the Circadian Locomotor Rhythm Induced by Pigment-Dispersing Factor in the Cricket Gryllus bimaculatus en-subtitle= kn-subtitle= en-abstract= kn-abstract=Pigment-dispersing factors (PDFs) are octadeca-peptides widely distributed in insect optic lobes and brain. In this study, we have purified PDF and determined its amino acid sequence in the cricket Gryllus bimaculatus. Its primary structure was NSEIINSLLGLPKVLNDA-NH2, homologous to other PDH family members so far reported. When injected into the optic lobe of experimentally blinded adult male crickets, Gryllus-PDF induced phase shifts in their activity rhythms in a phase dependent and dose dependent manner. The resulted phase response curve (PRC) showed delays during the late subjective night to early subjective day and advances during the mid subjective day to mid subjective night. The PRC was different in shape from those for light, serotonin and temperature. These results suggest that PDF plays a role in phase regulation of the circadian clock through a separate pathway from those of other known phase regulating agents. en-copyright= kn-copyright= en-aut-name=SingaravelMuniyandi en-aut-sei=Singaravel en-aut-mei=Muniyandi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=FujisawaYuko en-aut-sei=Fujisawa en-aut-mei=Yuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HisadaMiki en-aut-sei=Hisada en-aut-mei=Miki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SaifullahASM en-aut-sei=Saifullah en-aut-mei=ASM kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TomiokaKenji en-aut-sei=Tomioka en-aut-mei=Kenji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Department of Physics, Biology and Informatics, Faculty of Science, Research Institute of Time Studies, Yamaguchi University affil-num=2 en-affil= kn-affil=Suntory Institute for Bioorganic Research affil-num=3 en-affil= kn-affil=Suntory Institute for Bioorganic Research affil-num=4 en-affil= kn-affil=Department of Physics, Biology and Informatics, Faculty of Science, Research Institute of Time Studies, Yamaguchi University affil-num=5 en-affil= kn-affil=Department of Physics, Biology and Informatics, Faculty of Science, Research Institute of Time Studies, Yamaguchi University en-keyword=pigment-dispersing factor kn-keyword=pigment-dispersing factor en-keyword=circadian rhythm kn-keyword=circadian rhythm en-keyword=crickets kn-keyword=crickets en-keyword=phase shifts kn-keyword=phase shifts en-keyword=phase response curve kn-keyword=phase response curve END start-ver=1.4 cd-journal=joma no-vol=37 cd-vols= no-issue=2 article-no= start-page=67 end-page=72 dt-received= dt-revised= dt-accepted= dt-pub-year=2003 dt-pub=200303 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The role of helices of domain I for the insecticidal activity of Bacillus thuringiensis Cry4A toxin en-subtitle= kn-subtitle= en-abstract= kn-abstract=An active form of Cry4A is a heterodimer of the 20- and 45-kDa fragments that are derived from the 130-kDa Cry4A protoxin. To investigate the function of these two fragments, several deletion mutants were constructed and expressed in E.coli as the GST (glutathione-S-transferase) fusion proteins. The results of the bioassay against Culex pipiens larvae showed that the interaction of two fragments of Cry4A was necessary for the toxicity, and that the C-terminal 67 amino acids of the 20-kDa fragment corresponding to the helices ƒΏ4 and ƒΏ5 were involved in determining the insecticidal activity. Surprisingly the lack of helix ƒΏ5 did not affect the toxicity to C. pipiens, suggesting that the role of helix ƒΏ5 of Cry4A was different from that postulated in the case of Cry4A toxins. en-copyright= kn-copyright= en-aut-name=YamagiwaMasashi en-aut-sei=Yamagiwa en-aut-mei=Masashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=SakaiHiroshi en-aut-sei=Sakai en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= affil-num=1 en-affil= kn-affil=Department of Bioscience and Biotechnology Okayama University affil-num=2 en-affil= kn-affil=Department of Bioscience and Biotechnology Okayama University END start-ver=1.4 cd-journal=joma no-vol=38 cd-vols= no-issue=1-2 article-no= start-page=97 end-page=100 dt-received= dt-revised= dt-accepted= dt-pub-year=2004 dt-pub=200403 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The cytotoxicity of Bacillus thuringiensis subsp. coreanensis A2316 strain against the human leukemic T cell en-subtitle= kn-subtitle= en-abstract= kn-abstract=Bacillus thuringiensis subsp. coreanensis A2316 is a newly isolated strain from Yonakunijima Island in Japan. It produces the proteinaceous inclusion body (crystal) which has no insecticidal and hemolytic activities. When the crystal proteins were digested by proteinase K, they exhibited the strong cytotoxicity against human leukemic T cell, MOLT-4. The proteinase K-digested A2316 crystal proteins have little damage upon the cell membrane of MOLT-4, suggesting that the cell death of MOLT-4 was induced through a mechanism other than the colloid-osmotic swelling and cell lysis as caused by hitherto known B. thuringiensis crystal proteins. The 29-kDa polypeptide proved to be an active component of the proteinase K-digested A2316 crystal proteins. EC(50) of the purified 29-kDa polypeptide was 0.0579 ƒΚg/ml. The N-terminal amino acid sequence of the 29-kDa polypeptide was identical with that of p29 produced by B. thuringiensis A1519 strain and shared no significant homology with all the known proteins, suggesting that this polypeptide belong to a new family of B. thuringiensis crystal proteins. en-copyright= kn-copyright= en-aut-name=YamagiwaMasashi en-aut-sei=Yamagiwa en-aut-mei=Masashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HiraoTaichi en-aut-sei=Hirao en-aut-mei=Taichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KiyomiMasaaki en-aut-sei=Kiyomi en-aut-mei=Masaaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=AkaoTetsuyuki en-aut-sei=Akao en-aut-mei=Tetsuyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MizukiEiichi en-aut-sei=Mizuki en-aut-mei=Eiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=OhbaMichio en-aut-sei=Ohba en-aut-mei=Michio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SakaiHiroshi en-aut-sei=Sakai en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= affil-num=1 en-affil= kn-affil=Department of Bioscience and Biotechnology Okayama University affil-num=2 en-affil= kn-affil=Department of Bioscience and Biotechnology Okayama University affil-num=3 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University affil-num=4 en-affil= kn-affil=Biotechnology and Food Research Institute Fukuoka Industrial Technology Center affil-num=5 en-affil= kn-affil=Biotechnology and Food Research Institute Fukuoka Industrial Technology Center affil-num=6 en-affil= kn-affil=Graduate School of Agriculture Kyushu University affil-num=7 en-affil= kn-affil=Department of Bioscience and Biotechnology Okayama University END start-ver=1.4 cd-journal=joma no-vol=46 cd-vols= no-issue=5 article-no= start-page=761 end-page=763 dt-received= dt-revised= dt-accepted= dt-pub-year=2010 dt-pub=2010 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A novel method for screening peptides that bind to proteins by using multiple fluorescent amino acids as fluorescent tags en-subtitle= kn-subtitle= en-abstract= kn-abstract=We describe a new screening method for simultaneously detecting peptides that bind to a target protein by fluorescence obtained from fluorescent amino acid-modified peptides. en-copyright= kn-copyright= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=FutamiMidori en-aut-sei=Futami en-aut-mei=Midori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University affil-num=2 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University affil-num=3 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University END start-ver=1.4 cd-journal=joma no-vol=20 cd-vols= no-issue=20 article-no= start-page=5976 end-page=5978 dt-received= dt-revised= dt-accepted= dt-pub-year=2010 dt-pub=20101015 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Quantitative screening of EGF receptor-binding peptides by using a peptide library with multiple fluorescent amino acids as fluorescent tags en-subtitle= kn-subtitle= en-abstract= kn-abstract=EGF receptor-binding peptides could be found by a peptide screening method using fifteen fluorescent amino acids as fluorescent tags. Of 225 peptides, we found an 8-mer peptide containing a dipeptide unit, Y-F, which was the strongest binding peptide to the EGF receptor. en-copyright= kn-copyright= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YamamotoTakahiro en-aut-sei=Yamamoto en-aut-mei=Takahiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=FutamiMidori en-aut-sei=Futami en-aut-mei=Midori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University affil-num=2 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University affil-num=3 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University affil-num=4 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology, Okayama University en-keyword=Peptide kn-keyword=Peptide en-keyword=Peptide library kn-keyword=Peptide library en-keyword=Peptide screening kn-keyword=Peptide screening en-keyword=EGFR kn-keyword=EGFR en-keyword=Fluorescent amino acid kn-keyword=Fluorescent amino acid END start-ver=1.4 cd-journal=joma no-vol=123 cd-vols= no-issue=1 article-no= start-page=27 end-page=31 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20110401 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Renato Dulbecco: Dulbeccofs modified Eaglefs medium kn-title=Renato Dulbecco F ƒ_ƒ‹ƒxƒbƒR”|’n en-subtitle= kn-subtitle= en-abstract= kn-abstract=Œ»έCˆγŠw•ͺ–μ‚Μ‘½‚­‚ΜŒ€‹†‚Ν”|—{Χ–E‚β”|—{‘gD‚π—p‚’‚Δ‚Θ‚³‚κ‚Δ‚’‚ιD‚»‚κ‚η‚ΜŒ€‹†¬‰Κ‚ͺ•ρ‚³‚κ‚Δ‚’‚ι˜_•Ά‚́gή—Ώ‚Ζ•ϋ–@h‚̍€‚ŁCƒ_ƒ‹ƒxƒbƒR”|’n‚πŽg—p‚΅‚čזE‚π”|—{‚΅‚½‚Ζ‚’‚Α‚½‹LΪ‚π“Η‚ρ‚ΎŒ€‹†Ž‚͏­‚Θ‚­‚Θ‚’‚Ε‚ ‚λ‚€D‚ά‚½CŽΐΫ‚Ιƒ_ƒ‹ƒxƒbƒR”|’n‚πŽg—p‚΅‚½Œ€‹†Ž‚ΰ‚’‚ι‚ΖŽv‚€D‘‰ρ‚́C‚±‚Μ”|’n‚π•ρ‚΅‚½Dulbeccoi1975”NC‚ͺ‚ρƒEƒCƒ‹ƒX‚ΜŒ€‹†‚Ι‚ζ‚Α‚Δƒm[ƒxƒ‹άj‚Μ”g—–œδ‚ΜŒo—πC”ނ̐l–¬i‘½‚­‚̐l•¨‚ͺƒm[ƒxƒ‹ά‚πŽσάjC”ή‚ΜŒ€‹†ŠT—ͺ‚Θ‚Η‚Ι‚Β‚’‚ďЉ‚½‚’D‚»‚΅‚āC•MŽ‚Μƒ_ƒ‹ƒxƒbƒR”|’n‚Ι‘Ξ‚·‚ιŒ©‰π‚Ζ‚»‚Μ”|’n‚πŽg—p‚΅‚½ŒoŒ±‚Ι‚Β‚’‚ďq‚Χ‚ιD•MŽ‚ΜŒ‹˜_‚́C‰‘γΧ–E”|—{‚Ι‚Νƒ_ƒ‹ƒxƒbƒR”|’n‚ͺŠ©‚ί‚η‚κ‚ι‚±‚ƁC‚ά‚½C”|—{Χ–E‚π—p‚’‚Ă̓Ő«ŒŸ’θ‚ɍۂ΅‚ẮCŽg—p‚·‚ι”|’n‚Ι‚ζ‚Α‚ΔŒ‹‰Κ‚ͺˆΩ‚Θ‚ι‚±‚Ζ‚ͺ‚ ‚ι‚Μ‚Ε’ˆΣ‚·‚ι•K—v‚ͺ‚ ‚ι‚±‚Ζ‚Ε‚ ‚ιD en-copyright= kn-copyright= en-aut-name=NambaMasayoshi en-aut-sei=Namba en-aut-mei=Masayoshi kn-aut-name=“ο”g³‹` kn-aut-sei=“ο”g kn-aut-mei=³‹` aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=VŒ©Œφ—§‘εŠwC‰ͺŽRˆγŠwU‹»‰ο en-keyword=Renato Dulbecco kn-keyword=Renato Dulbecco en-keyword=ƒ_ƒ‹ƒxƒbƒR”|’n kn-keyword=ƒ_ƒ‹ƒxƒbƒR”|’n en-keyword=ƒm[ƒxƒ‹ά kn-keyword=ƒm[ƒxƒ‹ά en-keyword=Dulbecco‚̐l–¬ kn-keyword=Dulbecco‚̐l–¬ END start-ver=1.4 cd-journal=joma no-vol=100 cd-vols= no-issue= article-no= start-page=3 end-page=7 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20110201 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Purification and Characterization of l-Methionine Decarboxylase from Streptomyces sp. 590 kn-title=•ϊό‹ΫStreptomyces sp.590—R—ˆl-ƒƒ`ƒIƒjƒ“’E’YŽ_y‘f‚̐Έ»‚¨‚ζ‚ѐ«ŽΏŒŸ“’ en-subtitle= kn-subtitle= en-abstract= kn-abstract=L-Methionine decarboxylase [EC 4.1.1.57] catalyzes the decarboxylation of L-methionine and is a pyridoxal 5f-phosohate(PLP)-dependent enzyme. L-Methionine decarboxylase has been purified 630-fold by DEAE-Toyopearl 650M, Phenyl-Toyopearl 650M and Sephacryl S-300 column chromatographies from Streptomyces sp.590. The enzyme has a dimeric structure with identical subunits of Mr 60,000. This enzyme shows optimum activity at pH7.0 and 45‹C, and is stable between pH5.7 and pH9.0. L-Methionine decarboxylase has antitumor activity against RERF-LC-AI and HeLa cells. Ten N-terminal amino acid sequence of L-methionine decarboxylase was determined, and the sequence showed no homology with other reported proteins. en-copyright= kn-copyright= en-aut-name=MaemuraTomomi en-aut-sei=Maemura en-aut-mei=Tomomi kn-aut-name=‘O‘Ί’m”ό kn-aut-sei=‘O‘Ί kn-aut-mei=’m”ό aut-affil-num=1 ORCID= en-aut-name=UchitomiKumiko en-aut-sei=Uchitomi en-aut-mei=Kumiko kn-aut-name=“ΰ•x‹v”όŽq kn-aut-sei=“ΰ•x kn-aut-mei=‹v”όŽq aut-affil-num=2 ORCID= en-aut-name=KusakaChika en-aut-sei=Kusaka en-aut-mei=Chika kn-aut-name=“ϊ‰Ί’m kn-aut-sei=“ϊ‰Ί kn-aut-mei=’m aut-affil-num=3 ORCID= en-aut-name=InagakiJunko en-aut-sei=Inagaki en-aut-mei=Junko kn-aut-name=ˆξŠ_ƒŽq kn-aut-sei=ˆξŠ_ kn-aut-mei=ƒŽq aut-affil-num=4 ORCID= en-aut-name=TamuraTakashi en-aut-sei=Tamura en-aut-mei=Takashi kn-aut-name=“c‘Ί—² kn-aut-sei=“c‘Ί kn-aut-mei=—² aut-affil-num=5 ORCID= en-aut-name=SodaKenji en-aut-sei=Soda en-aut-mei=Kenji kn-aut-name=Ά‰E“cŒ’ŽŸ kn-aut-sei=Ά‰E“c kn-aut-mei=Œ’ŽŸ aut-affil-num=6 ORCID= en-aut-name=InagakiKenji en-aut-sei=Inagaki en-aut-mei=Kenji kn-aut-name=ˆξŠ_Œ«“ρ kn-aut-sei=ˆξŠ_ kn-aut-mei=Œ«“ρ aut-affil-num=7 ORCID= affil-num=1 en-affil= kn-affil=”_Œ|‰»ŠwƒR[ƒX affil-num=2 en-affil= kn-affil=”_Œ|‰»ŠwƒR[ƒX affil-num=3 en-affil= kn-affil=”_Œ|‰»ŠwƒR[ƒX affil-num=4 en-affil= kn-affil=‰ͺŽR‘εŠw‘εŠw‰@ˆγŽ•–ςŠw‘‡Œ€‹†‰Θ affil-num=5 en-affil= kn-affil=”_Œ|‰»ŠwƒR[ƒX affil-num=6 en-affil= kn-affil=‹ž“s‘εŠw affil-num=7 en-affil= kn-affil=”_Œ|‰»ŠwƒR[ƒX en-keyword=L-methionine decarboxylase kn-keyword=L-methionine decarboxylase en-keyword=pyridoxal 5f-phosohate kn-keyword=pyridoxal 5f-phosohate en-keyword=Streptomyces kn-keyword=Streptomyces en-keyword=decarboxylation of L-methionine kn-keyword=decarboxylation of L-methionine END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=12 article-no= start-page=2491 end-page=2499 dt-received= dt-revised= dt-accepted= dt-pub-year=1954 dt-pub=19541230 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=STUDIES ON THE TYROSINE LEVELS OF NON-HEAT PRECIPITATED, PROTEIN-LIKE MATERIALS OF HUMAN SERUM AND GASTRIC JUICE IN CANCER PATIENTS Chapter II Studies on the Tyrosine Levels of Non-heat Precipitated Protein-like Materials of Human Gastric Juice in Gastric Cancer Patients kn-title=ˆέŠΰŠ³Ž‚Μˆέ‰t‚ΙŠά‚ά‚κ‚ι‘Ξ”M”ρ‹ΓŒΕ•¨ŽΏŠe•ͺ‹ϋ’†‚Μƒ`ƒƒWƒ“—Κ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The protein-like materials of gastric juice consists of many kinds of non-heat precipitated mucous materials. By means of the quantitative analysis of the tyrosine levels contained in these materials and the isolated tyrosine level of gastric juice, the resolution of protein in cancer patients has been examined. In the case of the quantitative analysis of the tyrosine in total dissolved mucin and isolated tyrosine using trichlor acetic acid filtrate of gastric juice which was gathered after the subcutaneous injection of histamin, the tyrosine levels in total dissolved mucin in the cases of gastric cancer, gastric ulcer and gastritis were almost all the same. The isolated tyrosine levels in the cases of ulcer and gastritis, however, were 2-11 mg%, while that in the case of the gastric cancer was 9-26 mg%. It means the increase of isolated amino acid in gastric juice of cancer patients. According to Glass, Boyd and so forth, dissolved mucin was classified into mucoprotein and mucoproteose, and quantified the tyrosine levels of each of them. In the case of gastric cancer the tyrosine level of mucoprotein evidently decreased, but that of mucoproteose increased rather than that in the case of ulcer and gastritis. On the other hand the secretory volme of gastric juice showed clear decrease in patients with cancer. Consequently the tyrosine level of secretory mucoprotein (the tyrosine level of mucoprotein mg% ~ the secretory volume cc.) markedly decreased in the case of gastric cancer in comparison with the cases of ulcer and gastritis: that is, in the cases of gastric cancer the average level was 19 mg. in half an hour and 5 mg in 30 to 60 minutes after histamin injection, while in the cases of ulcer and gastritis the average levels were 201 mg and 107 mg respectively. The tyrosine levels of secretory mucoproteose did not show so clear distinction as that of mucoprotein Then fifteen cases of gastric cancer were classified into the following four groups. hydrochloric acid the tyrosine level of mucoprotein Group I (1 case) (+) normal Group II (3 cases) (-) normal Group III (7 cases) (-) decrease Group IV (4 cases) (-) little and impossible to quantitative analysis Comparing with clinical appearance, the anemia level was the highest in the group IV, which was followed by the group III, that of the group II was the lowest, and no anemia was noticed in the group I. In the view-points of the growth of cancer and indication of gastric resection, radical operations were possible in all the cases of group I, II and in the 4 cases of group III. On the other hand only the exploratory laparotomy was performed in the remaining 3 cases of group III and all the cases of group IV. According to Glass, Boyd, Rubinstein and so forth, mucoprotein is secreted from so-called gmucoid cellsh in the neck of gastric gland, and it is similar to Castle's intrinsic factor from the physical and physiological points of view. We often become aware in daily clinical experiences that the degree of anemia is often higher in the case of cancer of stomack than the other viscera such as breast, lung, esophagus, colon, rectum, and so forth. Therefore the deficiency of gastric mucoprotein may be related to the anemia level in gastric cancer patients. en-copyright= kn-copyright= en-aut-name=TaninoJyunzo en-aut-sei=Tanino en-aut-mei=Jyunzo kn-aut-name=’J–쏇‘’ kn-aut-sei=’J–μ kn-aut-mei=‡‘’ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”’Γ“cŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=12 article-no= start-page=2469 end-page=2476 dt-received= dt-revised= dt-accepted= dt-pub-year=1954 dt-pub=19541230 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=STUDIES ON THE QUANTITATIVE DETERMINATION OF PROCAINE, AND SERUM PROCAINE-ESTERASE IN SURGICAL REGION Chapter II The Distribution and the Output of Procaine and its Hydrolyzed Products kn-title=ŠO‰Θ—Μˆζ‚Ι‚¨‚―‚ι‰–Ž_ƒvƒƒJƒCƒ“‚Μ’θ—Κ‚ΖŒŒ΄ƒvƒƒJƒCƒ“EƒGƒXƒeƒ‰[ƒ[ ‘ζ2• ‰–Ž_ƒvƒƒJƒCƒ“i‹y‚Ρ‚»‚̐Ά‘Μ“ΰ•ͺ‰πŽY•¨j‚Μ‹zΎ•ΐ‚Ι”rŸ•‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The distribution or the output in blood or urine of procaine used on surgical operation and its hydrolyzed products has been examined by the method described above. On using 0.875g of procaine hydrochloride as local and splanchnic anesthesia, the blood total amount of the compounds which is showing a diazo-reaction on basis of procaine was maximum during the first two hours, and decreased gradully after then. On the other hand, during these wo hours while the laparotomy was performed, the amount of procaine and its hydrolyzed products showing a diazo-reaction which was discharged into urine was only 0.5% of the total amount having been used here. P-amino benzoic acid, which is one of its hydrolyzed products, almost changed into its acethyl compound. The amount of non-hydrolyzed procaine discharged into urine was about 1% of the total amount which has been used, and almost all of them was discharged within the first six hours after used. The ratio of non-acethyl type of p-amino benzoic acid to acethyl type of p-amino benzoic acid was 1:9 in the first six hours. After the six hours almost all the p-amino benzoic acid was discharged in acetyl ype. The total amount of p-amino benzoic acid and its acethyl type outpoured into urine was 50% of the amount having been used or more in the first six hours, and about 80% in twenty-four hours. en-copyright= kn-copyright= en-aut-name=TaninoJyunzo en-aut-sei=Tanino en-aut-mei=Jyunzo kn-aut-name=’J–쏇‘’ kn-aut-sei=’J–μ kn-aut-mei=‡‘’ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”’Γ“cŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=12 article-no= start-page=2461 end-page=2468 dt-received= dt-revised= dt-accepted= dt-pub-year=1954 dt-pub=19541230 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=STUDIES ON THE QUANTITATIVE DETERMINATION OF PROCAINE, AND SERUM PROCAINE-ESTERASE IN SURGICAL REGION Chapter I The Method of the Quanitative Determination of Procaine, and the Subsistence of Procaine-Esterase in Man kn-title=ŠO‰Θ—Μˆζ‚Ι‚¨‚―‚ι‰–Ž_ƒvƒƒJƒCƒ“‚Μ’θ—Κ‚ΖŒŒ΄ƒvƒƒJƒCƒ“EƒGƒXƒeƒ‰[ƒ[ ‘ζ1• ‰–Ž_ƒvƒƒJƒCƒ“‚Μ’θ—Κ‚¨‚ζ‚ΡƒvƒƒJƒCƒ“EƒGƒXƒeƒ‰[ƒ[‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The method of the quantitative determination of procaine that has been adopted here is the colorimetric one which modified Brodie's; that is, in place of N (1-naphthyl) ethylendiamine in Brodie's method, more excellent Tsuda's reagent, 1-(?-diethyl amino ethylamino) naphthalene dihydrochloride, has been used in ours, and buffer of boric acid (pH 9.0) has been used to separate procaine chemically from p-amino benzoic acid, hydrolyzed product of procaine produced by procainc-esterase in our method as well as in Brodie's. The procaine-esterase in 1 ml. of fresh human serum has the activity to he capable of hydrolyzing almost all the procaine hydrochloride 0.01 mg into p-amino benzoic acid and diethylamino ethanol in 4 to 5 minutes at shade temperature (10‹C). The older the serum is in an ice stockroom, the more this activity of procaine-esterase in serum decreases. Procaine-esterase is scarcely existing in blood-corpuscles and extracellular fluid, such as fluid in bulla by burns or pleural exudation. Futhermore it is said that the narcotic action of diethylamino ethanol produced by the hydrolysis of procaine is weaker than that of procaine. Therefore it is more suitable for procaine to be used as local anesthesia than intravenous one. en-copyright= kn-copyright= en-aut-name=TaninoJyunzo en-aut-sei=Tanino en-aut-mei=Jyunzo kn-aut-name=’J–쏇‘’ kn-aut-sei=’J–μ kn-aut-mei=‡‘’ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”’Γ“cŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=7-12 article-no= start-page=1399 end-page=1402 dt-received= dt-revised= dt-accepted= dt-pub-year=1955 dt-pub=19551231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Biochemical research on the experimental shock Part 6. On the mutual relation Found among the free aminoacids, especially glutamine, glutamic acid and ƒΑ-amino-butyric acid, in the cats-brains treated by insuline shock kn-title=ŽΐŒ±“IΥŒ‚‚̐Ά‰»Šw“IŒ€‹† ‘ζ6•Ρ ƒCƒ“ƒVƒ…ƒŠƒ“ΥŒ‚Ž{s”L”]‘‚Μ—V—£ƒAƒ~ƒmŽ_C“Α‚ΙƒOƒ‹ƒ^ƒ~ƒ“EƒOƒ‹ƒ^ƒ~ƒ“Ž_EƒΑ-ƒAƒ~ƒm—Ž_‚Μ‘ŠŒέŠΦŒW‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Have measured the amount of the free amino acids found with in cat's brain that has undergone the repeated Insulin shock, same as often used in curing psychotic patient: and which. according to paper chromatography, proved as follows: 1) As seen from the point of quality, no definite change whatever has happened. 2) Glutamic acid and ƒΑ-aminobutyric acid increased in quantity. 3) The ratio ƒΑ-aminobutyric acid to glutamic acid showed an increasing tendency. en-copyright= kn-copyright= en-aut-name=NishimonTakashi en-aut-sei=Nishimon en-aut-mei=Takashi kn-aut-name=Ό–δF kn-aut-sei=Ό–δ kn-aut-mei=F aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=7-12 article-no= start-page=1395 end-page=1398 dt-received= dt-revised= dt-accepted= dt-pub-year=1955 dt-pub=19551231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Biochemical research on the experimental shock Part 5. On the mutual relation Existing among the free amino acids, especially glutamine, glutamic acid and ƒΑ-aminobutyric acid, in the cat-brains treated by electric shocks kn-title=ŽΐŒ±“IΥŒ‚‚̐Ά‰»Šw“IŒ€‹† ‘ζ5•Ρ “dŒ‚Ž{s”L”]ι’‚Μ—V—£ƒAƒ~ƒmŽ_“Α‚ΙƒOƒ‹ƒ^ƒ~ƒ“EƒOƒ‹ƒ^ƒ~ƒ“Ž_EƒΑ-ƒAƒ~ƒm—Ž_‚Μ‘ŠŒέŠΦŒW‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The change of the free aminoacids in the cat-brains treated repeatedly by electric shocks was in vestigated. Both the systems of glutamic acid¨ƒΑ-aminobutyric acid and glutamic acid¨glutamine were influenced by the treatment. Especially the change in the former proved more remarkable than in the latter. In other words, glutamic acid decarboxylase has been activated and in the latter system, glutamine, has been formulated, though in a slight degree. These facts are considered to show certain rise in the brain function as results of E. C. T. en-copyright= kn-copyright= en-aut-name=NishimonTakashi en-aut-sei=Nishimon en-aut-mei=Takashi kn-aut-name=Ό–δF kn-aut-sei=Ό–δ kn-aut-mei=F aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=67 cd-vols= no-issue=7-12 article-no= start-page=1389 end-page=1393 dt-received= dt-revised= dt-accepted= dt-pub-year=1955 dt-pub=19551231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Biochemical research on the experimental shock Part 4. On the free amino acids contained in the brains of the normal cats kn-title=ŽΐŒ±“IΥŒ‚‚̐Ά‰»Šw“IŒ€‹† ‘ζ4•Ρ ³ν”L”]ι’‚Μ—V—£ƒAƒ~ƒmŽ_‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The present work was carried on by means of 2 dimensional paper chromatography. ewhere phenol and lutidine-collidine were used. The following results were obtained. 1) Aspartic acid, glutamic acid, glutamine, ƒΑ-aminobntpric acid and taurine were remarkably demonstrated, glycine and alanine moderately, while serine, ƒΐ-alanine, valine as well as leucine, slightly. 2) The free amino acids found in the brain are almost non-essential amino acids. 3) Certain fixed relation in quantitative ratio has been recognizable among glutamic acid, ƒΑ-aminobutyric acid and glutamine. en-copyright= kn-copyright= en-aut-name=NishimonTakashi en-aut-sei=Nishimon en-aut-mei=Takashi kn-aut-name=Ό–δF kn-aut-sei=Ό–δ kn-aut-mei=F aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=12 article-no= start-page=3175 end-page=3179 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19571231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Nitrogen Metabolism of the Brain Part 3 The Ammonia and Amino Acid Contents of the Rat Brain in case of the Injection of Chlorpromazine kn-title=”]‚Μ’‚‘f‘γŽΣ ‘ζ3• ƒNƒ[ƒ‹ƒvƒƒ}ƒaƒ““Š—^‘单‘l”]ι’ƒAƒ“ƒ‚ƒjƒA•ΐ‚ΙƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Ammonia of the rat brain in case of the injection of chlorpromazine was measured by Conway's microdiffusion method, and amino acid by the paperchromatography, and the following results have been obtained in the respective measurements. 1) Following the injection of chlorpromazine (50 mg/kg per once), the ammonia contents of the rat brain decreased to 0.34 mg. per cent. 2) In case of repeated injection of chlorpromazine (20 mg/kg every day), the ammonia contents of rat brain increased to 0.93 mg per cent, and the simultaneous decrease of glutamic acid was not found. 3) The amounts of ammonia formed by above mentioned brain slices were found to be 6.2 ƒΚM/g. for the first one hour, 7.1 ƒΚM/g in two hours, 9.3 ƒΚM/g. in three hours and 12.7 ƒΚM/g in four hours, respectively. Namely, in this case, the increase of ammonia formation was found during the first one hour and the decrease of it was found thereafter. en-copyright= kn-copyright= en-aut-name=KawaiKiyoshi en-aut-sei=Kawai en-aut-mei=Kiyoshi kn-aut-name=‰Νˆδ΄ kn-aut-sei=‰Νˆδ kn-aut-mei=΄ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=12 article-no= start-page=3135 end-page=3143 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19571231 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Biological Nature of Sh. sonnei Newly Isolated Part 2 On the Amino Acid Activity, Especially on the l-histidine decarboxylase Activity Induced by Sh. sonnei kn-title=V‚΅‚­•ͺ—£‚΅‚½Sh. sonnei‚̐Ά•¨Šw“I«σ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ2• Sh. sonnei‚Ι‚ζ‚ιaminoŽ_“Α‚Ιl-Histidine’E’YŽ_y‘fŠˆ«‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=There are just as many reports on the l-histidine decarboxylase activity of dysentery bacilli as those on the other bacteria. In the course of elucidation of the causative factors of violent food poisoning that occurred in the Okayama Prefecture in 1953, the author has made a report on the S-R dissociation and the virulency of Sh. sonnei in the Report 1. In the preaent report the biological nature of the virulent strain is described. 1) Sh. sonnei grow most rapidly in the medium whose optimal pH is 7.0-7.6, but when the pH falls under 4.3 the growth of bacteris is completely inhibited: and in addition, the peaks of the growth are correlated with changes in the pH of the medium. 2) In the cultures performed in test-tubes and flasks, R-type bacteia grew in the flask much better than S-type, while on the contrary in the test-tubes R-type bacteria growth was inferior to that of the S-type. 3) As for the growth of Sh. sonnei in soft gruel of rice malt, although the growth does not reach as high degree as observed in the peptone medium but it reaches the maximum growth within 12 hours. After 24-hour culture, however, the growth tends to decline. 4) On examining by paper chromatography and by Warburg's apparatus the l-histidine decarboxylase activity of the isolated strain that has been cultured in the media of peptone solution loaded with l-histidine and with additional various sugars, the excretion of carbon dioxide gas and the production of histamine are observable only in the cases where these bacteria are grown in the media containing additional glucose, maltose, or mannose. 5) Acetone powder of Sh. sonnei that has been cultured in the medium of glucose-peptone solution loaded with l-histidine possesses the l-histidine decarboxylase activity and acts specifically to the substrate. en-copyright= kn-copyright= en-aut-name=KobayashiRyosuke en-aut-sei=Kobayashi en-aut-mei=Ryosuke kn-aut-name=¬—Ρ—Η•γ kn-aut-sei=¬—Ρ kn-aut-mei=—Η•γ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=10 article-no= start-page=2497 end-page=2504 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19571031 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=The Characteristics of Staphylococci Isolated from Lesions in Human Mouth Part 2. On the Enzymatic Nature kn-title=Œϋo“ΰ•a‘ƒ‚ζ‚θ•ͺ—£‚΅‚½ƒuƒh[‹…‹Ϋ‚̐«σ‚Ι‚Β‚’‚Δ ‘ζ2• y‘f“I«σ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Similar to the previous report, 4 strains belonging to St. aureus and albus selected from the isolated bacteria, and each standard strain of aureus and albus and these bacteria passing through the animal were used as test bacteria. In the present experiment N-source requirement during the growth of these bacteria, and oxidation and production of amino acids by resting cells were studied; and glucose oxidation by growing bacteria and resting cells was compared. The results are as follows. 1. The N-source requirement in the case of albus compared with aureus in general tended to be greater but no significant difference could be recognized between standard strains and those passing through the animal or the isolated strains. 2. With respect to the oxidation and production of amino acids by resting cells, no distinct difference could be found either between standard strains and the bacteria passing through the animal or the isolated strains. 3. On comparing the glucose oxidation by growing bacteria and resting cells of each strain, during the process of glucose metabolism both the strains that passed through the animal and the isolated strains tend to show a weaker oxidation of pyruvate and of those that follow than the oxidation by our laboratory strains. en-copyright= kn-copyright= en-aut-name=YasuiRitsuya en-aut-sei=Yasui en-aut-mei=Ritsuya kn-aut-name=ˆΐˆδ—§œ\ kn-aut-sei=ˆΐˆδ kn-aut-mei=—§œ\ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=10 article-no= start-page=2485 end-page=2495 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19571031 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=The Characteristics of Staphylococci Isolated from Lesions in Human Mouth Part 1. Production of Hemolysins kn-title=Œϋo“ΰ•a‘ƒ‚ζ‚θ•ͺ—£‚΅‚½ƒuƒh[‹…‹Ϋ‚̐«σ‚Ι‚Β‚’‚Δ ‘ζ1• —nŒŒ“ΕŽYΆ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Using 4 strains considered to belong to Staphylococcus aureus and albus, selected from various bacteria isolated from the human mouth as test bacteria and one strain each of the standard-strain bacteria kept in our laboratory as the control, the author made comparative studies of the productivity of hemolysins of both growing bacteria and resting cells as well as investigated effects of various conditions on the production of hemolysins. The results are presented in the following. 1. By passage through the animal the productivity of hemolysin increases markedly in the case of standard strain of aureus while with albus the increase is only slight. 2. On the whole the productivity of hemolysin of the bacteria belonging to aureus strain is greater than those belonging to the albus, and No.2 and standard strain of aureus presents the hemolyzat the highest degree. 3. Nitrogen source is required in the production of hemolysins by resting cells, and the most effective one is pepton; while among amino acids, glutamate, aspartate, alanine and glycine are effective, but there is no amino acid known to be indispensable to the hemolysin production. Though hemolysin production does not occur when carbon source alone is added, when C-source is added in combination with N-source, the hemolysin production is enhanced. However, in the case of sugars, pH is lowered during the oxidation so there is a tendency to inhibit the hemolysin production. 4. Hemolysin production is inhibited by such metal ions as Fe(++), Mg(++), Co(++) and Cu(++), and it is likewise inhibited by monoiodoacetate, aureomycine, and chloromycetine. 5. Hemolysin produced by the resting cells are all unstable against heat with exception of standar strain of albus. en-copyright= kn-copyright= en-aut-name=YasuiRitsuya en-aut-sei=Yasui en-aut-mei=Ritsuya kn-aut-name=ˆΐˆδ—§œ\ kn-aut-sei=ˆΐˆδ kn-aut-mei=—§œ\ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=10 article-no= start-page=2477 end-page=2483 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19571031 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Nitrogen Metabolism of the Brain Part 2 The Ammonia and Amino Acid Contents of the Normal Rat Brain kn-title=”]‚Μ’‚‘f‘γŽΣ ‘ζ2•Ρ ³ν‘单‘l”]‘ƒAƒ“ƒ‚ƒjƒA•ΐ‚ΙƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Ammonia of the normal rat brain was measured by Conway's microdiffusion method, and amino acid by the paper chromatography, and the following data have been obtained in respective measurements. 1) In the case of the normal rat brain immersed in liquid nitrogen, the percentage of the contents of the following substances proved to be as follows: ammonia, 1.05 mg. per cent; glutamic acid, 140.9 mg. per cent; glutamine, 92.3 mg. per cent; and ƒΑ aminobutylic acid, 27.7 mg. per cent. 2) Trepanation and Enuculation of the Brain. Contents of the substances found in the brain frozen solid by dry ice acetone gave the following percentage; ammonia, 0.64 mg. per cent; glutamic acid, 159.9 mg. per cent; glutamine, 94.6 mg. per cent; and ƒΑ aminobutylic acid, 37.2 mg. per cent. 3) The amounts of ammonia formed by brain slices were found to be 5.1 ƒΚ M/g. for the first one hour, and 7.3 ƒΚ M/g. in two hours; and 13.8 ƒΚ M/g. in three hours, respectively. In this case, the decrease in the contents of glutamic acid and glutamine during the first two hours paralleled the increase in the contents of ammonia. en-copyright= kn-copyright= en-aut-name=KawaiKiyoshi en-aut-sei=Kawai en-aut-mei=Kiyoshi kn-aut-name=‰Νˆδ΄ kn-aut-sei=‰Νˆδ kn-aut-mei=΄ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=5 article-no= start-page=1299 end-page=1309 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19570531 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Experimental Study on the Acute Pancreatic Necrosis Chapter III. Paper chromatographic study on the change of free amino acid in the diseased pancreatic tissue kn-title=‹}«δX‘Ÿ‰σŽ€‚ΙŠΦ‚·‚ιŽΐŒ±“IŒ€‹† ‘ζ3• ΰhŽ†ƒNƒƒ}ƒgƒOƒ‰ƒtƒC‚Ι‚ζ‚ι‰σŽ€δX“ΰ—V—£ƒAƒ~ƒmŽ_‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The method of experiment of acute pancreatic necrosis in dogs was same as the described in Chapter I. The measurement was performed by the following method. Free amino acid in the pancreatic tissue was extracted by Dent's method. Chromatography was performed by a two-dimensional method using phenol and lutidincollidine solution, and phenol and butanol-acetic acid solution. The following results were obtained. In the normal pancreatic tissue the follwing 8 free amino acids were found: cystine, aspartic acid, glutamic acid, serine, glycine, lysine, arginine and threonine. In the diseased pancreatic tissues, leucine, valine and tyrosine were found besides the above eight. en-copyright= kn-copyright= en-aut-name=YakushijiMitsugu en-aut-sei=Yakushiji en-aut-mei=Mitsugu kn-aut-name=–ςŽtŽ›v kn-aut-sei=–ςŽtŽ› kn-aut-mei=v aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”’Γ“cŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=5 article-no= start-page=1247 end-page=1255 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19570531 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Metabolism of Mycobacterium tuberculosis III. Influences of various substrates on the oxygen consumption of Mycobacterium tuberculosis kn-title=Œ‹Šj‹Ϋ•¨ŽΏ‘γŽΣ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ3• Ž_‘fΑ”ο‚Ι‹y‚Ϊ‚·”Žν•¨ŽΏ‚ΜŒψ‰Κ en-subtitle= kn-subtitle= en-abstract= kn-abstract=With the use of intact cells aerated 10 hours and cell-free extracts of virulent and avirulent strains of both human and bovine tubercle bacilli, the oxygen consumption has been examined comparatively, adding each of the following substrates; sugars, organic acids and amino acids. The results are as follows: 1) Even after 10 hours aeration, in both of the human and bovine tubercle bacilli (including BCG), high endogenous oxygen consumption is observed, but when made into the cell-free extracts the rate of oxygen consumption of these bacteria decreases to 1/10 to 1/20 of that of the intact cells, thus making it quite easy to study the enzymic activities of bacteria to various substrates. 2) Sugars have very little influence on the oxygen consumption. 3) As for the influence of pyruvate on the oxygen consumption, it is makedly greater in H37Rv strain than H37Ra strain; but no significant difference can be seen between bovine 263 strain and BCG. 4) In the virulent strain, lactate increases the oxygen consumption markedly greater than in avirulent strain. 5) Although citrate increases the oxygen consnmption of BCG, it has no effect on that of other strains. 6) Amino acids have little accelerating effect on the oxygen consumption of bacteria except alnine which shows some effect in the case of H37Ra strain. en-copyright= kn-copyright= en-aut-name=OkaYoshikazu en-aut-sei=Oka en-aut-mei=Yoshikazu kn-aut-name=‰ͺD–œ kn-aut-sei=‰ͺ kn-aut-mei=D–œ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”’₯Ά•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=3 article-no= start-page=619 end-page=625 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19570331 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of Sh. flexneri II. Degradation of Cysteine and Methionine by Resting Cells kn-title=Sh. flexneri‚Ι‘‚·‚ι‹Ϋ‚ΜaminoŽ_‘γŽΣ ‘ζ2•Ρ ΓŽ~‹Ϋ‚Ι‚ζ‚ιcysteine, methionine‚Μ•ͺ‰π en-subtitle= kn-subtitle= en-abstract= kn-abstract=This experiment continued from Report I was conducted to examine the breakdown of cysteine and methionine by the resting cells of some strains of Sh. flexneri, and the following results were obtained. 1. When cysteine or methionine is added independently to the suspension of these resting cells, the breakdown of these amino acids does not occur, but when added with glucose it is analyzed: in the case of cysteine desulfydration occurs, H(2)S is formed and deamination can be observed, and in the case of methionine dethiomethylation occurs, CH(3)S-SCH(3) or relating compound is formed and deamination can be also observed. 2. Such breakdown of cysteine and methionine is accelerated by VB(6). 3. Compared with the ability of analysis of these S-containing amino acids of each strain, cysteine is more analyzed than methionine by the suitable cells as S source and methionine is more analyzed by the suitable cells as S source. en-copyright= kn-copyright= en-aut-name=UshidaSeishi en-aut-sei=Ushida en-aut-mei=Seishi kn-aut-name=‹“c½”V kn-aut-sei=‹“c kn-aut-mei=½”V aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=69 cd-vols= no-issue=3 article-no= start-page=611 end-page=618 dt-received= dt-revised= dt-accepted= dt-pub-year=1957 dt-pub=19570331 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of Sh. flexneri I. Relation between Growth and Amino-acid Metabolism kn-title=Sh. flexneri‚Ι‘‚·‚ι‹Ϋ‚ΜaminoŽ_‘γŽΣ ‘ζ1•Ρ ‹Ϋ‚Μ”­ˆη‚ΖaminoŽ_‘γŽΣ”\‚ΜŠΦŒW en-subtitle= kn-subtitle= en-abstract= kn-abstract=This experiment was conducted to examine the effect of aspartate and glutamate as N source and cysteine and methionine as S source for the growth of some strains belonging to Sh. flexneri, and the ability of amino acids syntheses by the resting cells of each strain. Thus the following results were obtained. 1. These organisms generally need aspartate or glutamate as N source and cysteine or methionine as S source, and many of them require nicotinamide. It is interesting that in each strain belonging to Sh. flexneri la, 3a, when growth medium is glutamate-cysteine system the organisms require nicotinamide, but when glutamate-methionine system, they do not require it. 2. If each of amino acids, such as aspartate, glutamate, alanine, is added to resting cells suspension independently, the formation of the other kinds of amino acids is little, but if each of them is added with glucose, the formation is great. At this time formed amino acids are mostly aspartate, glutamate and alanine, but to form valine from glutamate. and glucose with a strain in Sh. flex. 2b is peculiar. The above mentioned synthetic reaction of amino acids is remarkably accelerated by the addition of cysteine and methionine. en-copyright= kn-copyright= en-aut-name=UshidaSeishi en-aut-sei=Ushida en-aut-mei=Seishi kn-aut-name=‹“c½”V kn-aut-sei=‹“c kn-aut-mei=½”V aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=12-2 article-no= start-page=8401 end-page=8410 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591130 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Experimental Studies on Aspirin Allergy Part 1. Properties of Aspirin-protein kn-title=ƒAƒXƒsƒŠƒ“ƒAƒŒƒ‹ƒM[‚ΙŠΦ‚·‚ιŽΐŒ±“IŒ€‹† ‘ζ1• ƒAƒXƒsƒŠƒ“’`”’‚̐«σ‚ɏA‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Research on the biological activity of proteins has developed on the basis of studies carried on the chemical derivatives of proteins produced after a selective reaction between some chemical reagents and specific amino acid residual group in proteins. Following the techniques of G. C. Butler, C. R. Harington and M. E. Yuill (1940) the author produced aspirin-protein by chemically combining amino group of serum protein (ƒΓ-Amino group of lysine residual group) with aspirin, and carried out serological studies with this aspirin-protein, especially on the homogeneity between aspirin-protein and serum protein, and on the change in the property of proteins by the introduction of aspirin, and obtained the following results. 1. The free amino acid group consumed in the chemical combination of aspirin with protein can be confirmed by the ninhydrin reaction with reagent hydrindantin. 2. The viscosity of aspirin-protein is higher than that of normal serum protein. 3. And aspirin-protein has moved its isoelectric point towards the acidic side. 4. The aspirin group of aspirin-protein is possible to isolate and extract with toluene, after hydrolyzis aspirin-protein, and to be determined colorimeterically by Gerhardt's iron perchloride reaction. en-copyright= kn-copyright= en-aut-name=KimuraSusumu en-aut-sei=Kimura en-aut-mei=Susumu kn-aut-name=–Ψ‘ΊW kn-aut-sei=–Ψ‘Ί kn-aut-mei=W aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”ŒφO‰qΆŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=12-1 article-no= start-page=8003 end-page=8016 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591120 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acids Metabolism of Vibrio Cholera kn-title=ƒRƒŒƒ‰‹Ϋ‚ΜƒAƒ~ƒmŽ_‘γŽΣ‚Ι‚Β‚’‚Δ‘ζ1•Ρ ”­ˆη‚Ι‰—‚―‚ιNŒΉ, CŒΉ‚Μ‰e‹Ώ ‘ζ2•Ρ ΓŽ~‹Ϋ‚ΜƒAƒ~ƒmŽ_‘γŽΣ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Part I Effet of N and C-sources on Growth, of Vibrio Cholera Using the 3 strains of Vibrio cholera as test organisms, the original strain (INABA's strain), the intermediate variant strain (HIKOZIMA's strain) and the variant strain (OGAWA's strain), the author studied the effect of N- and C-sources on growth of these microorganisms, and obtained the following results. 1) The intermediate variant strain could be cultured by serial transfers on the madia containing glutamate as N-source and did not need other C-source or vitamins for its growth. While the other 2 strains, the original and the variant strains, could not be cultred successfully without the addition of yest extract into the media. Also these 2 strains could be cultured by serial transfers on the media containing peptone or casein hydrolysate instead of yeast extract, but failed to grow on the madia containing some dozen species of amino acids. 2) An acceleration effect on growth of the microorganisms was not so remarkable as in the case of other bacteria by the addition of C-sources except lactate. Although the addition of lactate showed the acceleration effect fairly well, contrarily that of glucose acted rather inhibitory on the growth. This evidence was possibly arisen from a decrease of pH of the media being caused by the oxidation of glucose. Part 2 Amino Acids Metabolism of Resting Cells Using the 3 strains of Vibrio chlera as in the preceding paper, Part I, the author studied on the oxidation and the convertion of amino acids by these microorganisms and obtained the following results. 1) All the microorganisms tested showed an accelerated O2-uptake at the expense of aspartic acid, glutamic acid, alanine and cysteine as substrate; but showed rather small O(2)-uptake at the other amino acids. 2) The optimum pH was found to be at about 7.0 on the oxidation of aspartic acid, glutamic acid and alanine. And the amino acids mentioned here were oxidized through deamination at the pH ranging 5.0 to 8.0. 3) The convertion of the amino acids from these mentioned above to the others was carried out more successfully by these microorganisms compared with the other. 4) Concerns about the fate of oysteine, it was supposed this amino acid would be undergone in the first place a deamination and a desulfhydration resulting in pyruvate. 5) Though tryptophanase activity was observed to some extent on the bacterial cells harvested from media just before test, the activity was raised adaptatively by the shaking of the resting cell suspension in the presence of tryptophane. However, this adaptation of microorganisms was inhibited with the addition of glucose. en-copyright= kn-copyright= en-aut-name=NakaoYasuo en-aut-sei=Nakao en-aut-mei=Yasuo kn-aut-name=’†”φ•Ϋ˜Y kn-aut-sei=’†”φ kn-aut-mei=•Ϋ˜Y aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=12-1 article-no= start-page=7989 end-page=8002 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591120 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Enzymatic Properties of Candida (I) kn-title=Candida‚̍y‘f“I«σ(I)‘ζ1• Ά‹Ϋ‚ΜO2Α”ο ‘ζ2• “€Œ‹Š£‘‡‹Ϋ‘Μ‹y‚ѐΆ‹Ϋ–›Σ’Šo‰t‚̐«σ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Part I O(2) Uptake of the Fresh Cells Using Candida albicans, C. krusei, C. parakrusei and C. tropicalis, the author studied the O(2) uptakes of the organisms at the expense of various carbon compounds or amino acids as substrate and the environmental factors to this. The following results were obtained. 1) The endogenous respiration of each strain was fairly high, and this respiration tended to decrease by the shaking of cell suspension without addition of substrates. Since prolonged shaking of the cell suspension may cause the inactivation of enzyme activity, the most advantage were given on the cell suspension that was previously shaken for 1-2 hrs. in order to study the enzymatic properties of the organisms. 2) Generally, the O(2) uptakes of Candida were high at the expense of glucose, acetate and citrte. Besides this, the O(2) uptake of C. albicans was also high at the expense of lactate and pyruvate. 3) As a whole, it could say that the greater enzyme activity was found on the cells of shorter cultivation compared with that of longer cultivation. And the organism cultured by shaking method showed more accelerated O(2) uptake at the oxidation of lactate, pyruvate and acetate than the cultured in still-standing method, this fact possibly implied that the metabolism of the former organism was carried out very satisfactory. Part II Properties of Freezing-dried Cell Preparation andFraction of Ground Fresh Cell As in the previous report, part I, using 4 strains of Candida, C. albicans, C. krusei, C. krusei, C. parakrusei and C. tropicalis, the author prepared the freezing-dried cell preparation and the fractions of ground fresh cells, studied O(2) uptake, catalase activity and action of inhibitors to these. The results obtained were following. 1) The freezing-dried cells showed an catalase activity in a same extent as the living cells, and a large amount of O(2) uptake with glucose, lactate, citrate and succinate; while a marked decrease of O(2) uptakes were found with pyruvate and acetate. 2) The supernatant fraction obtained from ground fresh cell at 40,000 rpm showed greater catalase activity, and greater oxidative capacity for pyruvate, acetate and citrate, and also accelerated endogenous respiration. On the other hand, the sediment separated by centrifugation at 40,000 rpm had no catalase activity; but revealed specifically much greater oxidative capacity for lactate and succinate. 3) It was confirmed that the action of inhibiter was more effective on the freezing-dried cells and the cell free extract than on the intact living cells; by an addition of KCN the O(2) upteak was not affected on the living cells, but was serionsly inhibited on the freezing-dried cells and the cell free extract. en-copyright= kn-copyright= en-aut-name=NishiEmiko en-aut-sei=Nishi en-aut-mei=Emiko kn-aut-name=ΌˆδΞ”όŽq kn-aut-sei=Όˆδ kn-aut-mei=Ξ”όŽq aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-2 article-no= start-page=7555 end-page=7557 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591030 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Decarboxylase in the Brain of Catfish kn-title=ƒiƒ}ƒY”]‚Ι‚¨‚―‚ιƒAƒ~ƒmŽ_’E’YŽ_y‘f‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=With the purpose to elucidate the mechanism of amino acids in the brain of fish, we have studied the glutamic decarboxylase in the fish brains following the techniques devise by Okumura et all, and examined the functions of various amino acid decarboxylase in the catfish brain. However, we did not find any decarboxylase activity in the catfish brain. en-copyright= kn-copyright= en-aut-name=YamadaTatsuo en-aut-sei=Yamada en-aut-mei=Tatsuo kn-aut-name=ŽR“c—΄—Y kn-aut-sei=ŽR“c kn-aut-mei=—΄—Y aut-affil-num=1 ORCID= en-aut-name=ImaiAkimasa en-aut-sei=Imai en-aut-mei=Akimasa kn-aut-name=‘ˆδΊ³ kn-aut-sei=‘ˆδ kn-aut-mei=Ί³ aut-affil-num=2 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ affil-num=2 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-2 article-no= start-page=7551 end-page=7554 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591030 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of the Brains. (IX) Transamination in the Chicken Brain kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(IX) ¬nƒjƒƒgƒŠ”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“ en-subtitle= kn-subtitle= en-abstract= kn-abstract=As a link in the studies on the transamination in the brain the author made a series of systematic studies on the transamination reactions in the brain of growing chick embryos, chicks at various stage of their growth, using 15 ƒΏ-amino acids, 4 diamino acids, cysteic acid and ƒΏ-ketoglutaric acid, and reported the result of such studies in previous papers. In the present experiment the transamination in the adult chicken brain was estimated in the similar way with the use of the same amino acids as in the previous experiment. 1. Of all the amino acids that showed the transaminase activity in the chick embryo and the chick brains at every developemental stage, lysine did not shows the activity in the adult chicken brains. 2. The grade of the transaminase activity of aspartic acid, isoleucine, ƒΐ-alanine and cysteic acid is lower than the peak of the same observed in the eavly stage of the chick brains. As for the other amino acids with an exception of alanine which showed the greatest activity in the adult chicken brain, none showed the activity surpassing the maximum shown in the chick brains at every developemental stage. 3. Spcaking relatively and biochemically at least in the chicken brain the transamination reactions in the chicken brains, especially to that of the mouse brain. en-copyright= kn-copyright= en-aut-name=YamadaTatsuo en-aut-sei=Yamada en-aut-mei=Tatsuo kn-aut-name=ŽR“c—΄—Y kn-aut-sei=ŽR“c kn-aut-mei=—΄—Y aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-2 article-no= start-page=7547 end-page=7550 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591030 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of the Brain. (VIII) Transamination in the Growing Chick Brains kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(VIII) ”­ˆη’†‚ΜƒjƒƒgƒŠƒqƒi”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“ en-subtitle= kn-subtitle= en-abstract= kn-abstract=With the growing chick brains take out at 9 different times during the period immediately after hatching to 40 days later the author studied the transaminase activity in these brains. The results are follows. 1. In ƒΏ-amino acids group aspartic acid, alanine, isoleucine, lencine, valine, norvaline. and tyrosine show the transaminase activity as the continuatoin from embryo and phenylalanine commences its transaminase activity. 2. In ƒΦ-amino acid group ƒΑ-aminobutylic acid, ƒΐ-alanine, ƒΐ-oxy-ƒΑ-aminobutylic acid, in the diamino acid group ornithine show the transaminase activity, while cysteic acid also as the continuation from the embryo shows the activity in every period examined. 3. Those amino acids showing the transaminase activity have individual peaks during the period from 4 to 10 days after hatching, and thereafter these amino acids take two different courses; namely, on that slightly falls folowing the respective Peak; and one that stays unchanged. en-copyright= kn-copyright= en-aut-name=YamadaTatsuo en-aut-sei=Yamada en-aut-mei=Tatsuo kn-aut-name=ŽR“c—΄—Y kn-aut-sei=ŽR“c kn-aut-mei=—΄—Y aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-2 article-no= start-page=7541 end-page=7545 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591030 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of the Brain. (VII) Transamination in the Growing Chick Embryo Brain kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(VII) ”­ˆη’†‚ΜƒjƒƒgƒŠγσ‘Ω”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Using the chick-embryo brains in its various developmental stage and by means of paperchromatography. the author studied transamination of 15 different ƒΏ-amino acids, 4 ƒΦ-amino acids, and ƒΏ-ketoglutaric acid from embryo-chemical standpoint, and investigated the relationship between the germination, functional activity and amino acid metabolism in the growing chick embryo brains. The following are the resnlts. 1. In the early stage of chick embryo brain which begins to germinate and develope, only a minimal transaminase activity can be recognized, but on about 11th embryonic day the activity grown gradually and consistently. On the day of hatching the transaminase activity is almost the same as in a chick. 2, Aspartic acid, alanine, isolecine, leucine, valine, norvalin, tyrosine, ornithine and ƒΐ-oxy-ƒΑ-amino acid, alanine, isoleuciue, leucine, valine, norvaline and cysteic acid immediately before hatching reaches about the same as found in other maturede animals such as mous (8) and dog (10). en-copyright= kn-copyright= en-aut-name=YamadaTatsuo en-aut-sei=Yamada en-aut-mei=Tatsuo kn-aut-name=ŽR“c—΄—Y kn-aut-sei=ŽR“c kn-aut-mei=—΄—Y aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-1 article-no= start-page=7317 end-page=7325 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591020 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Experimental Studies on the Influence of ƒΑ-Amino-ƒΐ-Hydroxybutyric Acid upon the Metabolism in Rabbit Brain after Cardiazol Convulsion Part 1. Infuence of ƒΑ-Amino-ƒΐ-Hydroxybutyric Acid upon the Free Amino Acid Nitrogen Content and Choliesterase Activity in Cerebral Cortex of Rabbits after Metrazcl Convulsion kn-title=CardiazolαzΉ‰Ζ“e”]‘‘γŽΣ‚Ι‚¨‚ζ‚Ϊ‚·ƒΑ-amino-ƒΐ-hydroxybutyric acid‚Μ‰e‹Ώ‚ΙŠΦ‚·‚ιŽΐŒ±“IŒ€‹† ‘ζ2• cardiazolαzΉ‰Ζ“e‘ε”]”ηŽΏ‚ΜNa. K.Šά—L—Κ‚Ι‚¨‚ζ‚Ϊ‚·ƒΑ-amino-ƒΐ-hydroxybutyric acid‚Μ‰e‹Ώ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The influence of GABOB and metrazol convulsion upon Na and K content in cerebral cortex of rabbits was studied, and then the influence of GABOB upon metrazol convulsion was investigated. Results were as follows: 1) The intravenous injection of 100 mg GABOB causes decrease of Na content in cerebral cortex of rabbits. The intravenous injection of 10 mg GABOB effects stronger than that of 100 mg in spite of its small dosis. 2) Na content in cerebral cortex of rabbits is increased by metrazol convulsion markedly in its first stadium and decreases from the medium of convulsion. But it does not recover completely even after 30 minutes from the end of the convulsion. 3) If 100 mg GABOB is given intravenously before metrazol convulsion, GABOB restraines the Na content, which had increased temporary by convulsion but which decreases with lapse of time. This level still continues 30 minutes after the end of convulsion. 4) The intravenous injection of 100 mg GABOB decreases markedly the K content in cerebral cortex of rabbits. In case of 10 mg intravenous injection the decrease is also found but it is weaker than that in the case of 100 mg. 5) K content in cerbral cortex of rabbits decreases markedly by metrazol convulsion and it does not recover completely still after 30 minutes from the end of convulsion. 6) The influnence of intravenous injection of 100 mg GABOB upon K content in cerebral cortex of rabbits during metrazol convulsion is seen neither in the first stadium nor the maximum stadium of convulsion but it accelerates the recover after the end of convulsion and let it recover to the normal level 30 minutes after the end of convulsion. en-copyright= kn-copyright= en-aut-name=KawakamiHirosi en-aut-sei=Kawakami en-aut-mei=Hirosi kn-aut-name=μγ—T kn-aut-sei=μγ kn-aut-mei=—T aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=11-1 article-no= start-page=7305 end-page=7315 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19591020 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Experimental Studies on the Influence of ƒΑ-Amino-ƒΐ-Hydroxybutyric Acid upon the Metabolism in Rabbit Brain after Cardiazol Convulsion Part 1. Infuence of ƒΑ-Amino-ƒΐ-Hydroxybutyric Acid upon the Free Amino Acid Nitrogen Content and Choliesterase Activity in Cerebral Cortex of Rabbits after Metrazcl Convulsion kn-title=CardiazolαzΉ‰Ζ“e”]‘‘γŽΣ‚Ι‚¨‚ζ‚Ϊ‚·ƒΑ-amino-ƒΐ-hydroxybutyric acid‚Μ‰e‹Ώ‚ΙŠΦ‚·‚ιŽΐŒ±“IŒ€‹† ‘ζ1• cardiazolαzΉ‰Ζ“e‘ε”]”ηŽΏ‚Μ—V—£ƒAƒ~ƒmŽ_Šά—L—Κ‚¨‚ζ‚ΡcholinesteraseŠˆ«’l‚Ι‚¨‚ζ‚Ϊ‚·ƒΑ-amino-ƒΐ-hydroxybutyric acid‚Μ‰e‹Ώ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The influences of GABOB (ƒΑ-amino-ƒΐ-hydroxybutric acid) upon free amino acid content and ChE activity in cerebral cortex of normal rabbits and those of rabbits after metrazol convulsin were studied. 1) After the injection of 100 mg GABOB, free amino acid nitrogen decreases temporary. Also after that of 10 mg GABOB, it shows a slight decrease. 2) During metrazol convulsion, free amino acid nitrogen content decreases markedly. 3) The intravenous injection of 100 mg GABOB before metrazol convulsion does not influence the variation of free amino acid nitrogen content during convulsion. Positively after that of 10 mg, the decrease of free amino acid nitrogen content by cardiazol convulsion is restrained slightly. 4) By intravenous injection of 100 mg GABOB., ChE activity is accelerated but by that of 10 mg, it is rather restrained. 5) During convulsion, the ChE activity in cerebral cortex decreases markedly. 6) In this case, if 100 mg GABOB is given intravenously before the intravenous injection of metrazol, in the maximum stadium of convulsion antagonistic function is fairly found, but in the latter stadium of convultion it is scarcely influenced. In case of 10 mg GABOB intravenous injection, ChE activity decreases by metrazol convulsion while it shows no sign of being influenced in the maximum stadium of convulsion. en-copyright= kn-copyright= en-aut-name=KawakamiHirosi en-aut-sei=Kawakami en-aut-mei=Hirosi kn-aut-name=μγ—T kn-aut-sei=μγ kn-aut-mei=—T aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=10-2 article-no= start-page=7057 end-page=7064 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590930 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Reduction of Bilirubin by Fecal Flora Part 2 A Study on the Bile Pigment Reduction Capacity of B. Coli Communis kn-title=’°“ΰΧ‹Ϋ‘p‚Ι‚ζ‚ιbilirubin‚ΜŠŒ³‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ2•Ρ E. coli communis‚Μ’_`F‘fŠŒ³”\‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The reduction of biliverdin was attempted by dissolving biliverdin in various media with the cultivation of either E. coli communis and the filtered bacterium solution. And the results were as follows. 1. In the non-proteinous media or in the pepton aqueous solution as the medium provided in the presence of 0.1% glucose biliverdin is reduced to bilirubin. However, in this instance the reduction takes place only at the central methin group but not at the vinyl group of side chain. 2. Even in the presence of pentose or hexose besides glucose similar result can be obtained, though the reduction power is less. In this instance sugar alone can not produce bilirubin. 3. Even under the condition unfavorable to the growth of bacterium, in the presence of glucose a small quantity of bilirubin is produced. 4. When various amino-acid sulfur compounds, l-ascorbic acid or egg-white exudate solution are added to the medium along with glucose, such addition only affected the velocity of the reduction and gave the same result. 5. In the presence of these substances added to various media a pigment possessing the characteristics of direct birect bilirubin has been isolated simultaneously. 6. Under the aerobic condition no effect other than the slowing-down of the reduction velocity can be recognized. 7. The factor that plays a role in the reduction of biliverdin is also excreted outside the body of E. coli communis during the culture. Furthermore, this factor is heatresistant and its reduction capacity shows itself only in the presence of glucose. en-copyright= kn-copyright= en-aut-name=MitsudaToshihiro en-aut-sei=Mitsuda en-aut-mei=Toshihiro kn-aut-name=Œυ“c—˜O kn-aut-sei=Œυ“c kn-aut-mei=—˜O aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζˆκ“ΰ‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=10-1 article-no= start-page=6455 end-page=6461 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590920 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Conversion of (14)C-Labelled Glucose to Amino Acid in Brain Tissue Part 2. Conversion of (14)C-labelled Glucose to Free Amino Acids in Brain Tissues of Idiopathic Epileptics and Rabbits with Latent Cerebral Local Anaphylaxis (LCLA) kn-title=”]‘‘gD‚Ι‚¨‚―‚ι(14)C-glucose‚ΜƒAƒ~ƒmŽ_‚Φ‚Μ“]‰»‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ2• ^³αŸα’Š³Ž‚Θ‚η‚тɐφέ«”]‹ΗŠƒAƒiƒtƒBƒ‰ƒLƒV[‰Ζ“e‚Μ”]‘‚¨‚Ι‚―‚ι(14)C-glucose‚ζ‚θ”]—V—£ƒAƒ~ƒmŽ_‚Φ‚Μ“]‰»‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The conversion of (14)C-labelled glucose to free amino acids by homogenates of brain tissues from idiopathic epleptics, non-epileptics as well as normal and LCLA rabbits was studied. 1) The turnover of (14)C-labelled glucose to free amino acids in idiopathic epileptic brain was more disturbed than that in non-epileptic brain: to asparagine and glutamine no significant differences wee seen, but especially to gluamic acid it was slightly disturbed and to aspartic acid and ƒΑ-aminobutyric acid they were intensively disturbed. In idiopathic epileptic brain no ƒΑ-aminobutyric acid was proved. 2) In one case of focal epileptic brain, the conversion of aspartic acid was more restrained in the focus than in the contrast part. 3) In LCLA rabbit brain, the turnover of free amino acids were generally more disturbed than that by normal one. To glutamine and asparagine no significant difference was found but to glutamic acid it was slightly and to ƒΑ-aminobutyric acid it was markedly disturbed. en-copyright= kn-copyright= en-aut-name=KurodaTakaaki en-aut-sei=Kuroda en-aut-mei=Takaaki kn-aut-name=•“c‘Έ–Ύ kn-aut-sei=•“c kn-aut-mei=‘Έ–Ύ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=10-1 article-no= start-page=6449 end-page=6454 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590920 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Conversion of (14)C-Labelled Glucose to Amino Acid in Brain Tissue Part 1. Conversion of (14)C-labelled Glucose to Amino Acid by Homogenate of human and Rabbits Brain kn-title=”]‘‘gD‚Ι‚¨‚―‚ι(14)C-glucose‚ΜƒAƒ~ƒmŽ_‚Φ‚Μ“]‰»‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ1• l”]‚¨‚ζ‚Ρ‰Ζ“e”]homogenate‚Ι‚ζ‚ι(14)C-glucose‚ζ‚θƒAƒ~ƒmŽ_‚Φ‚Μ“]‰»‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The conversion of (14)C-labelled glucose to free amino acids by homogenate of human and rabbit brain was studied by paper chromatgraphy and radioautography with X-ray film. Results were: 1) In both human and rabbit brain glutamine, asparagine, glutamic acid, aspartic acid and ƒΑ-aminobutyric acid with radioactivity were derived from (14)C-labelled glucose. Moreover in rabbit brain alanine was also derived. 2) In normal rabbit brain the derived amino acids were investigated. During short incubate time the precoursors such as aspartic acid and glutamic acid appeared quickly and intensively. Next asparagine and glutamine showed up and then alanine and ƒΑ-aminobutyric acid were proved. 3) It needed 24 hours for all free amino acids to appear sharply. 4) The optimal pH of this reaction was 8.5-9.0 en-copyright= kn-copyright= en-aut-name=KurodaTakaaki en-aut-sei=Kuroda en-aut-mei=Takaaki kn-aut-name=•“c‘Έ–Ύ kn-aut-sei=•“c kn-aut-mei=‘Έ–Ύ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=9-1 article-no= start-page=5651 end-page=5657 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590830 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of the Brain Part 2. Acitivity of Transaminase in the Brain kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζII• ”]ƒgƒ‰ƒ“ƒXƒAƒ~ƒi[ƒ[‚ΙŠΦ‚·‚ιŒ€‹† en-subtitle= kn-subtitle= en-abstract= kn-abstract=The activities of aspartic-glutamic transaminase and ƒΑ-aminobutyric-glutamic transaminate in rabbit brain and human cerbral cortex were determined. Results were: 1) In rabbit brain with repeated convulsions caused by electro-shocks for 10 days (5 times a day), the activity of aspartic-glutamic transaminase was less than that in the normal rabbit, but the activity of ƒΑ-aminobatyric-glutamic transaminase increased. However, there were no significant differences. 2) The activities of aspartic-glutamic transaminase and ƒΑ-aminobutyric-glutamic transaminase in the cerebral cortex of epilepsia decreased as compared with non-epileptic Iatients (patients with bran tumor), however no significant differencae were found. en-copyright= kn-copyright= en-aut-name=KokudoTadao en-aut-sei=Kokudo en-aut-mei=Tadao kn-aut-name=‘“y’‰’j kn-aut-sei=‘“y kn-aut-mei=’‰’j aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=9-1 article-no= start-page=5643 end-page=5650 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590830 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Amino Acid Metabolism of the Brain Part 1. Influences of Various Conditions on the Contents of Glutamic Acid and ƒΑ-Aminobutyric Acid in the Brain kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζI• ƒOƒ‹ƒ^ƒ~ƒ“Ž_‚¨‚ζ‚ΡƒΑ-ƒAƒ~ƒm—Ž_‚Μ”]“ΰ•Ο“‚ΙŠΦ‚·‚ιŒ€‹† en-subtitle= kn-subtitle= en-abstract= kn-abstract=The amount of glutamic acid and ƒΑ-aminobutyric acid in rabbit brain and human cerebral cortex under various conditions were determined by paperchromatography. The results were as follows: 1) After convulsion caused by electro-shock, the contents of glutamic acid and ƒΑ-aminobutyric acid in the rabbit brain markedly decreased. 2) In rabbits with repeated convulsions caused by electroshocks for 10 days (5 times every day), the contents of glutamic acid and ƒΑ-aminobutyric acid showed a decrease. 3) After the injection of chlorpromazin (50 mg/kg per time), the contents of glutamic acid and ƒΑ-aminobutyric acid decreased. 4) The contents of glutamic acid and ƒΑ-aminobutyric acid in the cerebral cortex of epilepsia were less as compared with non-epileptic patients (patients with brain tumor). en-copyright= kn-copyright= en-aut-name=KokudoTadao en-aut-sei=Kokudo en-aut-mei=Tadao kn-aut-name=‘“y’‰’j kn-aut-sei=‘“y kn-aut-mei=’‰’j aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=9-1 article-no= start-page=5513 end-page=5517 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590830 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Free Amino Acids in the Brains. X Free Amino Acids and Related Substances in the Brain of Hen (Gallus domesticus) and Common Turtle (Geoclemys Reevesii) in Active Stage kn-title=”]‚Μ—V—£ƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ(X) ƒjƒƒgƒŠ”]CŠˆ“ŠϊƒNƒTƒKƒ”]‚Μ—V—£ ƒAƒ~ƒmŽ_‚¨‚ζ‚Ρ‚»‚ΜŠΦ˜A•¨ŽΏ en-subtitle= kn-subtitle= en-abstract= kn-abstract=From the comparative biochmical standpoint the author isolated and estimated free amino acids and related substances in the brains of Hen (Gallus domesticus) and Common turtle (Geoclemys reevesii) in active stage, and obtaind the following results: 1. The free amino acid pattern of the hen brain is on the whole similar to that of the albino rat brain, containing X(3) and X(6) but no urea. 2. The free amino acid pattern of the common turtle brain reveals characteristics as the cold blooded animal and contains markedly greater volumes of aspartic acid, glutamic acid, glycine, alanine, glycerophosphoethanolamine, phosphoethanolamine and taurine as compared with those in the hibernating turtle. As for the volum of ƒΑ-amino butyric acid there can be recognized no difference between the brain of active turtle and that of hibernating one. en-copyright= kn-copyright= en-aut-name=AoyamaTatsuya en-aut-sei=Aoyama en-aut-mei=Tatsuya kn-aut-name=ΒŽR’B–η kn-aut-sei=ΒŽR kn-aut-mei=’B–η aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=8-2 article-no= start-page=5087 end-page=5099 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590815 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Influences of Various Non-Essential Amino Acids on the Hematopoiesis Part 3. Influences of hydroxyproline on the erythropoiesis and application of this substance in hypoplastic anemia kn-title=ŠeŽν‰ΒŒ‡ƒAƒ~ƒmŽ_‚̍œ‘‘’ŒŒ‹@”\‚Ι‹y‚Ϊ‚·‰e‹Ώ ‘ζ3• ƒqƒhƒƒLƒVƒvƒƒŠƒ“‚ΜΤŒŒ‹…Œn‘’ŒŒ‚Ι‹y‚Ϊ‚·‰e‹Ώ•ΐ‚ɍĐΆ•s—ǐ«•nŒŒ‚Φ‚Μ‰ž—p en-subtitle= kn-subtitle= en-abstract= kn-abstract=The author studied influences of hydroxyproline on erythropoiesis by means of rabbit bone-marrow tissue culture and also used this substance in the treatment of hypoplastic anemia; and obtained the following results. 1. In loading hydroxyproline directly to rabbit bone marrow tissue culture(fluid medium), no significant change can be recognized in the number of erythrocytes and reticulocytes. As for the change in hemoglobin (Hb) content with addition of this substance, Hb decreases in the control whereas it increases slightly when added in a high concentration. 2. In the tissue culture of the bone marrow (cover slip method) obtained from sternum of 5 cases of hypoplastic anemia loaded with hydroxyproline, the tissue growth has been accelerated in three cases. The two unaffected cases turned out to be of panmyelophthisis type. 3. Of 5 cases with hypoplastic anemia to three cases daily intravenous injection of 150-300 mg hydroxyproline was given. As the result all of them showed an increase the leucocyte count, but the erythrocyte count and Hb increased only in one case and it proved to be ineffective in other two cases. en-copyright= kn-copyright= en-aut-name=LeeBin Zen en-aut-sei=Lee en-aut-mei=Bin Zen kn-aut-name=—›•q‘R kn-aut-sei=—› kn-aut-mei=•q‘R aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”•½–Ψ“ΰ‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=8-2 article-no= start-page=5079 end-page=5085 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590815 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Influences of Various Non-Essential Amino Acids on the Hematopoiesis Part 2. Influences of various non-essential amino acids on the wandering velocity of pseudoeosinophils in rabbit bone-marrow tissue culture kn-title=ŠeŽν‰ΒŒ‡ƒAƒ~ƒmŽ_‚̍œ‘‘’ŒŒ‹@”\‚Ι‹y‚Ϊ‚·‰e‹Ώ ‘ζ2• ‰Ζ“eœ‘‘gD”|—{‚Ι‰—‚―‚ι‹UDŽ_‹…—V‘–”\‚Ι‹y‚Ϊ‚·ŠeŽν‰ΒŒ‡ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ en-subtitle= kn-subtitle= en-abstract= kn-abstract=By adding polytamine, by-product of cow's milk hydrolysis and 10 different non-essential amino acids as already mentioned in Part 1 directly to rabbit bone-marrow tissue culture, the author studied influences of these amino acids on the wandering velocity of mature pseudoeosinophils in the bone marrow. 1. Polytamine and by-product of cow's milk caseous hydrolysis do not at all affect the wandering velocity of pseudoeosinophils. 2. Hydroxyproline, proline and cysteine, when added in a high concentration, all accelerate the wandering velocity but they do not at all affect the wandering velocity when added in a low concentration. Moreover, cysteine has been found to accelerate the wandering velocity even 12 hours after the start of culture just as much as at the third hour. The other two accelerate the wandering velocity markedly up to the sixth hour of culture. 3. Glutamic acid accelerates the wandering velocity slightly. 4. Arginine, serine, glycine, tyrodine, aspartic acid, and ornithine have neither accelerating action nor inhibitory action on the wandering velocity of pseudoeosinophils. en-copyright= kn-copyright= en-aut-name=LeeBin Zen en-aut-sei=Lee en-aut-mei=Bin Zen kn-aut-name=—›•q‘R kn-aut-sei=—› kn-aut-mei=•q‘R aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”•½–Ψ“ΰ‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=8-2 article-no= start-page=5067 end-page=5078 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590815 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Influences of Various Non-Essential Amino Acids on the Hematopoiesis Part 1. Influences of various non-essential amino acids on leucocyte growth in rabbit bone marrow tissue culture kn-title=ŠeŽν‰ΒŒ‡ƒAƒ~ƒmŽ_‚̍œ‘‘’ŒŒ‹@”\‚Ι‹y‚Ϊ‚·‰e‹Ώ ‘ζ1• ‰Ζ“eœ‘‘gD”|—{‚Ι‰—‚―‚ι”’ŒŒ‹…‘Ά‚Ι‹y‚Ϊ‚·ŠeŽν‰ΒŒ‡ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ en-subtitle= kn-subtitle= en-abstract= kn-abstract=By adding such substances as polytamine, a derivative of amino acids (a by-product of caseous fermentation of cow's milk), by-product of cow's milk caseous hydrolysis with acid, and ten different non-essential amino acids contained in polytamine directly to rabbit bonemarrow tissue culture, the author estimated the tissue growth area and cell density and studied influences of these substances on the production of the leucocyte series in the bone marrow. The results are as follows. 1. Both polytamine and by-product of cow's milk caseous hydrolysis with acid can increase the growth area slightly more than the control only in a high concentration. 2. With addition of L-hydroxyproline in the concentration from 0.1 to 0.0001 per cent, this substance in any concentration within this range increases the growth area and cell density 2-3 times that of the control, showing the greatest values among amino acids used in the present experiment. 3. Both L-proline and L-arginine in high concentration increase the growth area twice the control, and slightly at low concentration. 4. L-Cysteine, L-tyrodine, and DL-serine each in a proximal concentration can only increase the growth area slightly more than the control. 5. L-Aspartic acid and L-glutamic acid, glycine, and L-ornithine show hardly any significant difference as compared with the control. en-copyright= kn-copyright= en-aut-name=LeeBin Zen en-aut-sei=Lee en-aut-mei=Bin Zen kn-aut-name=—›•q‘R kn-aut-sei=—› kn-aut-mei=•q‘R aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”•½–Ψ“ΰ‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=8-1 article-no= start-page=4679 end-page=4685 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590810 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=The Influence of Amino Acid upon Cholinestrase Activity in the Brain Part 2. The influence of amino acid upon cholinestrase acitivity in the cerebral cortex of rabbits with latent local anaphylaxis kn-title=”]ƒRƒŠƒ“ƒGƒXƒeƒ‰[ƒ[‚Ι‚¨‚ζ‚Ϊ‚·ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ2• φέ«”]‹ΗŠƒAƒiƒtƒCƒ‰ƒLƒV[‰Ζ“e‘ε”]”ηŽΏ‚ΜƒRƒŠƒ“ƒGƒXƒeƒ‰[ƒ[Šˆ«‚Ι‚¨‚ζ‚Ϊ‚·ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Various influecnes of amino acids upon ChE activity in the brain of latent cerebral local anaphylactic rabbits (LCLA rabbits) and normal ones were studied. 1) The ChE activity in the brain of LCLA rabbits accelerated stronger than that of normal rabbits, but after adding amino acid it fell to normal value or less. 2) By addition of amino acid the ChE activity redueces both in normal rabbits and LCLA rabblts, but it was more marked in the latter. 3) The intensity of the straint of amino acids for the ChE activity is the largest by glutamine to normal rabbits. Aspartic acid, ƒΑ-aminobutyric acid, asparagine, glutamic acid follows. To LCLA rabbits glutamine effects excellent and then asparagine, ƒΑ-aminobutyric acid, aspartic acid, glutamic acid. 4) If the density of adding amino acid solution is changed in glutamine and glutamic acid, they show a maximum straint at M/10. In glutamine it shows respectable straint even at M/200. 5) The influence of glutamine upon the ChE activity in the brain of LCLA rabbits showed straint even at M/100 and at M/200 it became equal with the level of normal rabbit brain. This is found to be almost the same value as the deficient value of amino nitrogen in LCLA rabbit brain. en-copyright= kn-copyright= en-aut-name=YamaguchiMichiya en-aut-sei=Yamaguchi en-aut-mei=Michiya kn-aut-name=ŽRŒϋœNΖ kn-aut-sei=ŽRŒϋ kn-aut-mei=œNΖ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=8-1 article-no= start-page=4671 end-page=4677 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590810 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=The Influence of Amino Acid upon Cholinestrase Activity in the Brain Part 1. The influence of amino acid upon cholinesterase activity in cerebral cortex of idiopathic epileptics kn-title=”]ƒRƒŠƒ“ƒGƒXƒeƒ‰[ƒ[‚Ι‚¨‚ζ‚Ϊ‚·ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ1• ^«αŸα’‘ε”]”ηŽΏ‚ΜƒRƒŠƒ“ƒGƒXƒeƒ‰[ƒ[Šˆ«‚Ι‚¨‚ζ‚Ϊ‚·ƒAƒ~ƒmŽ_‚Μ‰e‹Ώ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The ChE activity of cerebral cortex resected from idiopathic epileptics and non-epilcptics and the influences of glutamic acid, glutamine, aspartic acid, asparagine, ƒΑ-aminobutyric acid and ƒΑ-amino-ƒΐ-hydrooxybutyric acid were studied. 1) In the brain of idiopathic epileptics, ChE activity is more markedly accelerated than that in the brain of non-epileptics. 2) By adding the amino-acids mentioned above, restraint occurs in each case. The intensity of these restraint for the non-epilcptic brain has the order of asparagine, glutamine, ƒΑ-amino-ƒΐ-hydroxybutyric acid, glutamic acid, aspartic acid, glutamine. ƒΑ-aminobutyric acid, while that for idiopathic epileptic brain it is asparagine, aspartic acid, glutamine, ƒΑ-amino-ƒΐ-hydroxybutyric acid, glutamic acid ƒΑ-aminobutyric acid. 3) The reduction of ChE activity by adding amino acids is found in each case of epileptic and non-epileptic brain. But it is not so definite in epileptic brain as in none-epileptic brain. 4) The ascending of ChE activity in idiopathic epileptic brain is regarded to be caused by the reduction of free amino acids in idiopathic brain. en-copyright= kn-copyright= en-aut-name=YamaguchiMichiya en-aut-sei=Yamaguchi en-aut-mei=Michiya kn-aut-name=ŽRŒϋœNΖ kn-aut-sei=ŽRŒϋ kn-aut-mei=œNΖ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”‘ζ1iw“ΰjŠO‰Θ‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=6-2 article-no= start-page=3187 end-page=3191 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590515 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Free Amino-Acids in the Brain (X) on the free amino-acids and related substances in various parts in the mature and foetal human brains kn-title=”]‚Μ—V—£ƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ(X) ¬nƒqƒg”]‚¨‚ζ‚Ρƒqƒg‘ΩŽ™”]Še•”ˆΚ‚Ι‚¨‚―‚ι—V—£ƒAƒ~ƒmŽ_‚¨‚ζ‚Ρ‚»‚ΜŠΦ˜A•¨ŽΏ‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract=With the brains of foetuses on the fifth gravid month and on the eighth gravid month the author isolated and estimated the free amino-acids and related substances at various parts of these brains by means of the ion-exchange chromatography, and conducted the same investigations with the normal tissues surrounding the tumor of the frontal lobe in the brain of an 18-year-old patient with tubosclerosis. 1. The quantities of free amino-acids in the foetal brains differ significantly from those in the mature human brain. 2. In the foetal brain, with some variations according to sites, amino-acids such as taurine, phosphoethanolamine, glycine and alanine, generally tend to decrease with the advance in age, while glutaminic, aspartic and ƒΑ-amino-butyric acids tend to increase. 3. In either one of the foetal brains ƒΑ-amino-butyric acid is detected in a largest quantity in the hypothalamus. 4. Cystathionine which is often detected in a large quantity in the human brain after birth can hardly be recognized in the foetal brain. kn-abstract=ƒCƒIƒ“ŒπŠ·ƒNƒƒ}ƒgƒOƒ‰ƒtƒC[‚π‚ΰ‚Ώ‚’‚Δ,”DP5ƒPŒŽ‚¨‚ζ‚Ρ8ƒPŒŽ‘ΩŽ™”]‚ΜŠe•”ˆΚ‚Θ‚η‚Ρ‚Ι18ΛŒ‹ί«d‰»ΗŠ³Ž”]‚Μ‘O“ͺ—t,Žξα‡Žό•Σ‚Μ”ρŒ‹ί«‘gD‚Ι‚Β‚’‚Δ—V—£ƒAƒ~ƒmŽ_‚¨‚ζ‚Ρ‚»‚ΜŠΦ˜A•¨ŽΏ‚πŠeX•ͺ—£’θ—Κ‚΅‚½. 1) ”DP5ƒPŒŽ‚Θ‚€‚Ρ‚Ι8ƒPŒŽ‘ΩŽ™”]‚Ɛ¬nƒqƒg”]‚π”δŠr‚·‚ι‚Ζ,‘ΩŽ™”]‚ƐΆŒγƒqƒg”]‚Ζ‚ΜŠΤ‚Ι‚Ν‚»‚Μ—V—£ƒAƒ~ƒmŽ_—Κ‚Ι‚©‚Θ‚θ’˜–Ύ‚ȍ·ˆΩ‚π”F‚ί‚ι. 2) ‘ΩŽ™”]‚Ε‚Ν,”]Še•”ˆΚ‚Ζ‚ΰ‚Ι‘½­‚Μ—αŠO‚Ν‚ ‚ι‚ͺ,Άˆη‚Μ’iŠK‚Ι‚Β‚κ‚Δƒ^ƒEƒŠƒ“,ƒzƒXƒzƒGƒ^ƒmƒ‰ƒ~ƒ“,ƒOƒŠƒVƒ“,ƒAƒ‰ƒjƒ““™‚ΝŒΈ­‚΅,ƒOƒ‹ƒ^ƒ~ƒ“Ž_,ƒAƒXƒpƒ‰ƒMƒ“Ž_, ƒΑ-ƒAƒ~ƒm—Ž_“™‚Ν‘‰Α‚·‚ιŒXŒό‚πŽ¦‚΅‚Δ‚’‚ι. 3) ƒΑ-ƒAƒ~ƒm—Ž_‚Ν‘ΩŽ™”]‚Μ‚’‚Έ‚κ‚Ε‚ΰ,Ž‹°‰Ί•”‚ōō‚’l‚πŽ¦‚΅‚½. 4) ΆŒγ‚Μƒqƒg”]‚Ε‘½—Κ‚Ι‘ͺ’θ‚³‚κ“Ύ‚ιƒVƒXƒ^ƒ`ƒIƒjƒ“‚Ν‘ΩŽ™”]‚Ι‚Ν‚Ω‚Ζ‚ρ‚Η‘Άέ‚΅‚Θ‚’. en-copyright= kn-copyright= en-aut-name=FukaiNobuhiro en-aut-sei=Fukai en-aut-mei=Nobuhiro kn-aut-name=[ˆδ‰„_ kn-aut-sei=[ˆδ kn-aut-mei=‰„_ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=6-1 article-no= start-page=2997 end-page=3007 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590501 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Tryptophan Metabolism of Bacteria II. Tryptophan metabolism of B. typhosus kn-title=Χ‹Ϋ‚ΜƒgƒŠƒvƒgƒtƒ@ƒ“‘γŽΣ‚ΙŠΦ‚·‚ιŒ€‹† ‘ζ2• ’°ƒ`ƒtƒX‹Ϋ‚ΜƒgƒŠƒvƒgƒtƒ@ƒ“‘γŽΣ en-subtitle= kn-subtitle= en-abstract= kn-abstract=It is well known that tryptophan is necessary for the growth of B. typhosus. The anthor analyzed this requirement of tryptophan from the point of view of the nutritive requirement of B. typhosus. The results were as follows: 1) Of ten strains, seven strains require tryptophan for growth, three strains do not. 2) S-58 which requires tryptophan utilizes no amino acid singlely as N source and S-60 which does not require tryptophan utilizes singlely glutamic acid, cystine and aspartic acid. Glutamic acid is especially utilized by S-60. 3) S-58 does not grow in the absence of tryptophan and grows scarcely in the absence of cystine and grows a little in the absence of aspartic acid. On the contrary S-60 grows in the presence or absence of tryptophan and grows scarcely in the absence of cystine and grows a little in fhe absence of aspartic acid. 4) Tryptophan has the effect on growth at 10(-8) Mol. From this and the fact that above mensioned the strain which requires tryptophan could not grow in the absence of tryptophan tryptophan is fouud to be ggrowth factorh fot the strain. 5) Indole is found to be approximately equally effective as tryptophan. 6) Indole acetic acid and skatole is more weak effective compared with indole. en-copyright= kn-copyright= en-aut-name=InadaMinoru en-aut-sei=Inada en-aut-mei=Minoru kn-aut-name=ˆξ“cŽΐ kn-aut-sei=ˆξ“c kn-aut-mei=Žΐ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””χΆ•¨Šw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1651 end-page=1654 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Metabolism of Amino Acids in the Brain (VI) Studies on the Transamination in the Brain of Frog (Rana Nigromacnlata) kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(VI) ƒgƒmƒTƒ}ƒKƒGƒ‹”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=With a view to make biochemical comparison the author studied the transamination of various amino acids, and ƒΏ-ketoglutaric acid in the brain of frog (Rana Nigromaculata), and compared these results with those obtained with the brains of mice, catfish and common turtle. 1. ƒΏ-amino acids in the brains of frog show on the whole a lower transaminase activity as compared with those in the brains of catfish and mice, but glycine and methionine that show no transamination other than a slight transaminase activity in the brain of catfish reveal a fairly active transamination in the frog brain. 2. In the frog brain threonine and tyrodine are not at all transaminated. 3. Phenylalanine is not at all transaminated in the brains of cold-blooded animals. 4. Of ƒΦ-amino acids with an exception of ƒΑ-aminobutyric acid in the brain of catfish, all other show a low transaminase activity in the brains of cold-blooded animals. 5. The degree of the enzyme activity of di-amino acids does not differ to any marked extent in different animals. en-copyright= kn-copyright= en-aut-name=ImaiAkimasa en-aut-sei=Imai en-aut-mei=Akimasa kn-aut-name=‘ˆδΊ³ kn-aut-sei=‘ˆδ kn-aut-mei=Ί³ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1647 end-page=1650 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Metabolism of Amino Acids in the Brain (V) Studies on the Transamination in the Brain of Common Turtle (Clemmys japonica) kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(V) ƒCƒVƒKƒ”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=From the comparative biochemical viewpoint the author studied the transamination of 16 different ƒΏ-amino acids, 4 of ƒΦ-amino acids, di-amino acids, cysteic acid and ƒΏ-ketoglutaric acid in the brain of common turtle (Clemmys japonica); and obtained the following results: 1. When compared with the brains of catfish and mice the brain of common turtle reveals generally a lower transaminase activity, especially it is markedly low in the case of alanine, leucine, valine, norvaline, tyrodine, ƒΑ-aminobutyric acid, and cysteic acid. 2. The same as in the case of the brain of catfish no transaminase activity of phenylalanine can be observed in the brain of common turtle; and also methionine and glycine are not transaminated just as in the mouse brain. 3. In the brain of common turtle ƒΐ-alanine is not at all transaminated. 4. Arginine shows the transaminase activity in the brain of common turtle. en-copyright= kn-copyright= en-aut-name=ImaiAkimasa en-aut-sei=Imai en-aut-mei=Akimasa kn-aut-name=‘ˆδΊ³ kn-aut-sei=‘ˆδ kn-aut-mei=Ί³ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1641 end-page=1645 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Metabolism of Amino Acids in the Brain (IV) Studies on the Transamination in the Brain of Catfish (Parasilurus asotus) kn-title=”]‚ΜƒAƒ~ƒmŽ_‘γŽΣ(IV) ƒiƒ}ƒY”]‚Ι‚¨‚―‚ιƒgƒ‰ƒ“ƒXƒAƒ~ƒl[ƒVƒ‡ƒ“‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=Using the brains of catfish (Parasilurus asotus) and mouse as the test materials and with the aid of paper chromatography the author compared the transamination of 16 kinds of ƒΏ-amino acids, 4 different ƒΦ-amino acids, 4 di-amino acids, cysteic acid and ƒΏ-ketoglutaric acid and studied the possibility of the amino acid metabolism in the brain of catfish. The results are as follows: 1. The brains of catfish demonstrate generally a high transaminase activity as compared with the brains of mice, especially the activity of aspartic acid, alanine, leucine and threonine is marked. 2. Glycine and methionine that are not transaminated in the mouse brain demonstrate their activity in the brain of catfish. 3. Phenylalanine is not transaminated in the brain of catfish. en-copyright= kn-copyright= en-aut-name=ImaiAkimasa en-aut-sei=Imai en-aut-mei=Akimasa kn-aut-name=‘ˆδΊ³ kn-aut-sei=‘ˆδ kn-aut-mei=Ί³ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1635 end-page=1639 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Free Amino Acids in the Brains IX The Influence of Hypophysectomy on the Free Amino Acids and Related Compounds in the Brain of Rat kn-title=”]‚Μ—V—£ƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ(IX) ‰Ί‚‘Μ™‘œ‚Μ”]—V—£ƒAƒ~ƒmŽ_‚¨‚ζ‚Ρ‚»‚ΜŠΦ˜A•¨ŽΏ‚Ι‚¨‚ζ‚Ϊ‚·‰e‹Ώ en-subtitle= kn-subtitle= en-abstract= kn-abstract=With the aid of the chromatography on the 150 x 0.9 cm column of Dowex 50-x4 the author estimated 20 kinds of free amino acids and their related compounds isolated from the brain of the hypophysectomized Rattus; and obtained the following results: 1. A marked decrease has been observed in the metabolically active amino acid groups such as aspartic acid, glutamic acid, ƒΑ-aminobutyric acid, alanine and serine. 2. Likewise a decrease can clearly be noticed in the content of N-acetylaspartic acid that exists specifically in the brain and in the content of phosphor containing amine, glycerophosphoethanolamine as well as in the glutathione content, the only peptide which has been successfully estimated. 3. In the sutdy of 6 indispensable amino acids such as threonine, isoleucine, leucine, lysine, histidine, and arginine no noteworthy changes can be recognized. en-copyright= kn-copyright= en-aut-name=NishiokaHirosuke en-aut-sei=Nishioka en-aut-mei=Hirosuke kn-aut-name=Ό‰ͺ”Ž•γ kn-aut-sei=Ό‰ͺ kn-aut-mei=”Ž•γ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1629 end-page=1634 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on the Free Amino-Acids in the Brains VIII On the Free Amino-Acids and Related Compounds in the Brain of Dog kn-title=”]‚Μ—V—£ƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ(VIII) ƒCƒk”]Še•”ˆΚ‚Μ—V—£ƒAƒ~ƒmŽ_‚¨‚ζ‚Ρ‚»‚ΜŠΦ˜A•¨ŽΏ en-subtitle= kn-subtitle= en-abstract= kn-abstract=By means of the chromatography on the 150~0.9cm column of Dowex 50-x 4 author estimated various free amino-acids and related compounds isolated from such sites of the brains of dog as the cerebral cortex, white matter, cerebellar hemisphere, caudal nucleus, thalamus, hypothalamus, medulla oblongata, and cerebellar worm; and obtained the following results: 1. It has been found that the content of glutamic acid is low in the white matter, hypothalamus and medulla oblongata, while the aspartic acid content is likewise somewhat low in the white matter but quite high in the medulla oblongata. The content of ƒΑ-aminobutyric acid is markedly high in the hypothalamus, followed by that in the cerebellar worm, but in is low in the white matter. 2. The contents of cystathionin and glutathion are markedly high in the thalamus and hypothalamus. 3. The content of N-acetylaspartic acid is high in the cerebral cortex while it is low in the white matter and medulla oblongata. en-copyright= kn-copyright= en-aut-name=FukaiNobuhiro en-aut-sei=Fukai en-aut-mei=Nobuhiro kn-aut-name=[ˆδ‰„_ kn-aut-sei=[ˆδ kn-aut-mei=‰„_ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=71 cd-vols= no-issue=4-1 article-no= start-page=1591 end-page=1594 dt-received= dt-revised= dt-accepted= dt-pub-year=1959 dt-pub=19590325 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Studies on Free Amino Acids in the Brain Part VII Estimation of Free Amino Acids in the Human Brain by the Ion-Exchange Chromatography kn-title=”]‚Μ—V—£ƒAƒ~ƒmŽ_‚Ι‚Β‚’‚Δ (VII) ƒCƒIƒ“ŒπŠ·ƒNƒƒ}ƒgƒOƒ‰ƒtƒC[‚Ι‚ζ‚ιƒqƒg”]—V—£ƒAƒ~ƒmŽ_‚Μ’θ—Κ en-subtitle= kn-subtitle= en-abstract= kn-abstract=By means of the ion-exchange chromatography free amino acids and their related compounds in the fresh brain tissue have been estimated. As for the test material, pieces of the cerebral cortex sectioned at the time of the surgical operation in the patients with cerebral tumor or with epilepsy are used. As the result we have obtained the following notworthy differences when compared with the results previously reported by us concerning several species of mammalians. They are: (1) In the human brain there is a conciderable amount of cystathionine. Namely, 19.4 to 43.2mg/100g wet weight, and this substance is far smaller or not recognizable in other animals; and (2) the amounts of ƒΑ-aminobutric acid and taurine are a good deal smaller than animals. Moreover, in general the pattern of the free amino acids in the human cerebral cortex may be said to resemble closely that in dog. en-copyright= kn-copyright= en-aut-name=YunoueShigeru en-aut-sei=Yunoue en-aut-mei=Shigeru kn-aut-name=“’”Vγ–Ξ kn-aut-sei=“’”Vγ kn-aut-mei=–Ξ aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”_ŒoΈ_ˆγŠw‹³ŽΊ END start-ver=1.4 cd-journal=joma no-vol=101 cd-vols= no-issue=7-8 article-no= start-page=687 end-page=698 dt-received= dt-revised= dt-accepted= dt-pub-year=1989 dt-pub=19890831 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Purification and characterization of HBs antigen from hepatoma huGK-14 cell line kn-title=ƒqƒg”|—{ŠΜŠΰΧ–E‚ΜŽYΆ‚·‚ιHBsRŒ΄‚̐Έ»•ϋ–@‚Ζ‚»‚Μ•¨—‰»Šw“I“Α«‚Ι‚Β‚’‚Δ en-subtitle= kn-subtitle= en-abstract= kn-abstract=HBs antigen was purified from the culture fluid of hepatoma huGK-14 cell line and its physico-chemical properties were studied. The purification consists of following steps: concentration of culture fluid by membrane filtration, affinity column chromatography (anti-HBs monoclonal antibody column and anti-human serum albumin antibody column), and ultracentrifugation (isopycnic centrifugation in CsCl density gradient and rate zonal centrifugation on sucrose gradient). Highly purified (purity>99%) HBs antigen was isolated with an overall yield of about 40%. The HBs antigen showed uniform spherical particles (diameter: 23.2}2.9nm) and had a specific gravity of 1.20g/cm3. The purified HBs antigen yielded, in SDS-PAGE (under reducing conditions), four protein bands with apparent molecular weights of 22,000 and 26,000 (the two major bands), and 44,000 and 47,000. The two proteins of molecular weights of 26,000 and 47,000 are likely to be glycosylated, as these were several fold reduced when the cells were cultured in the presence of Tunicamycin. Amino acid analysis, Edman degradation, carboxypeptidase digestion, and ultraviolet absorption spectrum indicated that the HBs antigen from hepatoma cells is very similar to that derived from human plasma. en-copyright= kn-copyright= en-aut-name=OdaMunehiro en-aut-sei=Oda en-aut-mei=Munehiro kn-aut-name=¬“c@G kn-aut-sei=¬“c kn-aut-mei=@G aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”ŠΰŒΉ•a—Šw‹³ŽΊ en-keyword=Hepatoma kn-keyword=Hepatoma en-keyword=Cell culture kn-keyword=Cell culture en-keyword=HBs antigen kn-keyword=HBs antigen en-keyword=Purification kn-keyword=Purification en-keyword=Characterization kn-keyword=Characterization END start-ver=1.4 cd-journal=joma no-vol=101 cd-vols= no-issue=5-6 article-no= start-page=659 end-page=672 dt-received= dt-revised= dt-accepted= dt-pub-year=1989 dt-pub=198906 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Expression and purification of the HIV-1 env gene products in Escherichia coli kn-title=HIV-1ƒGƒ“ƒxƒ[ƒv’`”’ŽΏ‚Μ‘ε’°‹Ϋ‚Ε‚Μ”­Œ»‚ƐΈ» en-subtitle= kn-subtitle= en-abstract= kn-abstract=To purify the HIV-1 envelope protein with antigenic reactivity, the Pvu II-Bgl II fragment of the HIV-1 env gene, from the Pvu II site to the second Bgl II site, encoding the carboxyl terminal 180 amino acids of the viral surface protein (SU, gp120) was molecularly cloned in Escherichia coli strain HB101 using protein A expression-shuttle vector pRIT5. The pRIT5 contains the protein A gene, encoding the secretion signal and IgG binding domain of protein A with the upstream promoter and the downstream multicloning sites, as well as the two replication sites for Escherichia coli and Staphylococcus aureus. A fused protein with the molecular weight of about 55 kilodaltons was produced, which showed the same reactivity as the native protein A against rabbit serum IgG on Western blotting analysis. Most of the fused protein in the periplasmic space was degraded, while the complete fused protein inside the cells was recovered as an insoluble protein. The fused protein was solubilized with sodium dodecylsulfate (SDS), partially purified by IgG sepharose affinity chromatography, and completely purified by SDS-polyacrylamide gel electrophoresis. The quantity of the expressed fused protein was estimated about 1% of the total proteins. The purified fused protein contained 516 amino acids with Mr54, 976, consisting of 305 amino acids of the IgG binding domain of protein A, 5 amino acids derived from polylinker, a carboxyl terminal 180 amino acids of the HIV-1 envelope surface protein gp120, and 26 amino acids derived from the pUC19 sequence. en-copyright= kn-copyright= en-aut-name=ZhangBo en-aut-sei=Zhang en-aut-mei=Bo kn-aut-name=’£”g kn-aut-sei=’£ kn-aut-mei=”g aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”ŠΰŒΉŒ€‹†Ž{έΆ‰»Šw•”–ε en-keyword=HIV-1 kn-keyword=HIV-1 en-keyword=ƒGƒ“ƒxƒ[ƒv’`”’ŽΏ kn-keyword=ƒGƒ“ƒxƒ[ƒv’`”’ŽΏ en-keyword=”­Œ»ƒxƒNƒ^[ kn-keyword=”­Œ»ƒxƒNƒ^[ en-keyword=Protein A kn-keyword=Protein A en-keyword=IgG-SepharoseƒJƒ‰ƒ€ kn-keyword=IgG-SepharoseƒJƒ‰ƒ€ END start-ver=1.4 cd-journal=joma no-vol=102 cd-vols= no-issue=11-12 article-no= start-page=1373 end-page=1384 dt-received= dt-revised= dt-accepted= dt-pub-year=1990 dt-pub=199012 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Effect of N-methyl-D-aspartic acid on amino acids levels in the mouse brain kn-title=N-ƒƒ`ƒ‹-D-ƒAƒXƒpƒ‰ƒMƒ“Ž_”]ŽΊ“ΰ“Š—^‚Μƒ}ƒEƒX”]“ΰƒAƒ~ƒmŽ_‚Φ‚Μ‰e‹Ώ en-subtitle= kn-subtitle= en-abstract= kn-abstract=The naturally-occurring dicarboxylic amino acids, L-glutamate (Glu) and L-aspartate, are the principal neurotransmitter candidates for excitatory synaptic transmission in vertebrate central nervous systems. The receptors activated by these amino acids are classified by their most selective and potent agonists, i.e., N-methyl-D-aspartic acid (NMDA), kainic acid, and quisqualic acid. In this study, I examined the effects of NMDA on behavior, electroencephalogram (EEG), and brain amino acids levels after intraventricular injection in mice. When NMDA was intraventricularly injected into mice, running fits were observed 10-30 seconds after injection, followed by a sedative phase and returned to a normal behavior within 15-20 minutes after injection. In the EEG, middle voltage fast waves were observed 10-20 seconds after injection, followed by EEG suppression for a few minutes and the appearance of high voltage slow waves 4-5 minutes after injection. About 20 minutes after the injection the EEG was normal. No spike discharge was observed during this observation. Glu levels increased in the hippocampus during running fits, while GABA levels decreased in the cerebellum and hippocampus before running fits, and increased in the cerebellum 10 minutes after NMDA injection. The taurine level decreased in the striatum before running fits. All amino acids observed recovered to control levels 60 minutes after NMDA injection. These results indicate that the NMDA-induced running fits are not accompanied by spike discharges in the EEG, and are related to Glu and GABA neurons. en-copyright= kn-copyright= en-aut-name=OkamuraYuji en-aut-sei=Okamura en-aut-mei=Yuji kn-aut-name=‰ͺ‘Ί—TŽi kn-aut-sei=‰ͺ‘Ί kn-aut-mei=—TŽi aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””]‘γŽΣŒ€‹†Ž{έ‹@”\Ά‰»Šw•”–ε en-keyword=N-methyl-D-aspartic acid kn-keyword=N-methyl-D-aspartic acid en-keyword=excitatory amino acids kn-keyword=excitatory amino acids en-keyword=inhibitory amino acids kn-keyword=inhibitory amino acids en-keyword=running fits kn-keyword=running fits en-keyword=mouse EEG kn-keyword=mouse EEG END start-ver=1.4 cd-journal=joma no-vol=102 cd-vols= no-issue=3-4 article-no= start-page=419 end-page=434 dt-received= dt-revised= dt-accepted= dt-pub-year=1990 dt-pub=199004 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=A physiological study of guanidinoethanesulfonic acid-induced seizure activity kn-title=Guanidinoethanesulfonic acid‚Μ‚―‚’‚κ‚ρ—U”­μ—p‚ΙŠΦ‚·‚鐢—Šw“IŒ€‹† en-subtitle= kn-subtitle= en-abstract= kn-abstract=The effect of guanidinoethanesulfonic acid (amidino-taurine, GES) on electrocorticograms (ECoG) and the effects of a 4-aminobutanoic acid (GABA)-agonist and anticonvulsants on the GES-induced ECoG changes were studied. Sporadic spike discharges began 2-5 min after GES (1 ƒΚ mol) application to the pia mater of the sensorimotor cortex of a rat with a frequency of 5-10 spike discharges/min. Spike discharges in the ECoG of the opposite cerebral hemisphere were observed 60 min after the onset of the spike discharges. The spike discharges lasted until the end of recording after 3 h. The GES-induced spike discharges were completely suppressed with the supplementation of taurine (1-3 ƒΚ mol) on the pia mater following the completion of the spike discharges. GABA (1 ƒΚ mol) and its agonists, (3 R)-(-)-4-amino-3-hydroxybutanoic acid and muscimol (10 nmol) when applied topically, also suppressed the GES-induced spike discharges. While diazepam (DZP) (10 mg/kg, i. p.), which acts as a functional GABA-agonist, showed suppressive effects on the GES-induced spike discharges following their completion, phenobarbital (PB) (20 mg/kg, i. p.), which also acts as a functional GABA-agonist, increased the frequency and voltage of spike discharges. While ethosuximide (100 mg/kg, i. p.) showed weak suppressive effects on spike discharges, intraperitoneal injection of either valproate (200 mg/kg) or phenytoin (25 mg/kg) did not affect them. These findings suggest that neurotransmission or neuromodulatory effects of taurine might participate in the induction mechanism of GES-induced seizure activities, and that GABA and DZP receptors might play a role in the mechanism which suppresses GES-induced seizures. en-copyright= kn-copyright= en-aut-name=TodaHiroko en-aut-sei=Toda en-aut-mei=Hiroko kn-aut-name=ŒΛ“c—mŽq kn-aut-sei=ŒΛ“c kn-aut-mei=—mŽq aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•””]‘γŽΣŒ€‹†Ž{έ‹@”\Ά‰»Šw•”–ε en-keyword=guanidinoethanesulfonic acid kn-keyword=guanidinoethanesulfonic acid en-keyword=guanidino compound kn-keyword=guanidino compound en-keyword=experimental seizures kn-keyword=experimental seizures en-keyword=GABA-receptor antagonist kn-keyword=GABA-receptor antagonist en-keyword=anticonvulsant kn-keyword=anticonvulsant END start-ver=1.4 cd-journal=joma no-vol=102 cd-vols= no-issue=3-4 article-no= start-page=347 end-page=356 dt-received= dt-revised= dt-accepted= dt-pub-year=1990 dt-pub=199004 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Antagonistic activity of L-histidine to the methamphetamine-induced hyperactivity in mice kn-title=Methamphetamine‚Ι‚ζ‚ιƒ}ƒEƒX‚Μ‰^“—Κ‘‰Α‚Ι‘Ξ‚·‚ιL-histidine‚̝hRμ—p‚ΙŠΦ‚·‚ιŒ€‹† en-subtitle= kn-subtitle= en-abstract= kn-abstract=The mechanism by which L-histidine inhibits the methamphetamine (MAP)-induced hyperactivity was examined in mice. L-histidine (1g/kg, i. p.) significantly reduced the locomotor hyperactivity induced by MAP (1mg/kg, i. p.) in either experiment using Open field or Animex apparatus, although L-histidine did not affect the locomotor activity of control mice. L-histidine elevated brain levels of histamine and tele-methylhistamine. Pretreatment with ƒΏ-fluoromethylhistidine, a histidine decarboxylase inhibitor, suppressed both behavioral and biochemical effects of L-histidine. Metoprine, a histamine-N-methyltransferase inhibitor, increased brain histamine level and suppressed the MAP-induced locomotor hyperactivity. Although L-histidine (1g/kg) markedly changed brain tyrosine levels, this amino acid alone had no significant influence on the monoamine metabolism. It did not modify the change in the monoamine metabolism induced by MAP (1mg/kg), either. The inhibition by L-histidine of the MAP-induced hyperactivity was observed in the mice pretreated with para-chlorphenylalanine or atropine. These results suggest that central histaminergic systems may have an inhibitory influence on the MAP-induced locomotor hyperactivity. However, serotonergic and cholinergic mechanisms seem not to be involved in this phenomenon. en-copyright= kn-copyright= en-aut-name=SenohShin en-aut-sei=Senoh en-aut-mei=Shin kn-aut-name=–…”φa kn-aut-sei=–…”φ kn-aut-mei=a aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=‰ͺŽR‘εŠwˆγŠw•”–ς—Šw‹³ŽΊ en-keyword=methamphetamine kn-keyword=methamphetamine en-keyword=L-histidine kn-keyword=L-histidine en-keyword=histamine kn-keyword=histamine en-keyword=‰^“—Κ kn-keyword=‰^“—Κ END start-ver=1.4 cd-journal=joma no-vol=13 cd-vols= no-issue= article-no= start-page=1 end-page=5 dt-received= dt-revised= dt-accepted= dt-pub-year=1953 dt-pub=195312 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=PHOTOMETRIC DETERMINATION OF MAGNESIUM IN NATURAL WATERS en-subtitle= kn-subtitle= en-abstract= kn-abstract=Various colormetric methods for determining magnesium in natural waters have been studied, and the methods using 8-oxyquinolin, ammonium molybdate and titan yellow were studied most frequently(1)), following the studies on the interfering ions(2)). Brilliant yellow(3)), l-amino-2-naphthol-6-sulfonic acid(4)) and other new reagents were also used. E. D. T. A. was used, but the determination by using this reagent is not exact. Present reagent already reported by T. Ashizawa(5)), magneson II, is insoluble in water, soluble in alkali and hardly soluble in ethanol. In the existence of magnesium ion, the color of solution varies from pink-violet (in alkali) and orange (in ethanol) to blue-violet. This variation of color was evaluated photometrically by Shimadzu photoelectric spectrophotometer, and moreover the grades of interference by interfering ions were clarified. en-copyright= kn-copyright= en-aut-name=UmemotoShunji en-aut-sei=Umemoto en-aut-mei=Shunji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=CHEMICAL DIVISION, BALNEOLOGICAL LABORATORY, OKAYAMA UNIVERSITY END start-ver=1.4 cd-journal=joma no-vol=68 cd-vols= no-issue=11 article-no= start-page=1530 end-page=1536 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20076 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Serine racemases from barley, Hordeum vulgare L., and other plant species represent a distinct eukaryotic group: gene cloning and recombinant protein characterization en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Several D-amino acids have been identified in plants. However, the biosynthetic pathway to them is unclear. In this study, we cloned and sequenced a cDNA encoding a serine racemase from barley which contained an open reading frame encoding 337 amino acid residues. The deduced amino acid sequence showed significant identity to plant and mammalian serine racemases and contained conserved pyridoxal 5-phosphate (PLP)-binding lysine and PLP?interacting amino acid residues. The purified gene product catalyzed not only racemization of serine but also dehydration of serine to pyruvate. The enzyme requires PLP and divalent cations such as Ca2+, Mg2+, or Mn2+, but not ATP, whereas mammalian serine racemase activity is increased by ATP. In addition to the results regarding the effect of ATP on enzyme activity and the phylogenetic analysis of eukaryotic serine racemases, the antiserum against Arabidopsis serine racemase did not form a precipitate with barley and rice serine racemases. This suggests that plant serine racemases represent a distinct group in the eukaryotic serine racemase family and can be clustered into monocot and dicot types.

en-copyright= kn-copyright= en-aut-name=FujitaniYoshiyuki en-aut-sei=Fujitani en-aut-mei=Yoshiyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HoriuchiTerumi en-aut-sei=Horiuchi en-aut-mei=Terumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=ItoKazutoshi en-aut-sei=Ito en-aut-mei=Kazutoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SugimotoManabu en-aut-sei=Sugimoto en-aut-mei=Manabu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Plant Bioengineering Research Laboratories, Sapporo Breweries Ltd. affil-num=4 en-affil= kn-affil=Okayama University en-keyword=Hordeum vulgare L. kn-keyword=Hordeum vulgare L. en-keyword=Oryza sativa kn-keyword=Oryza sativa en-keyword=Gramineae kn-keyword=Gramineae en-keyword=Pyridoxal 5-phosphate kn-keyword=Pyridoxal 5-phosphate en-keyword=Serine racemase kn-keyword=Serine racemase en-keyword=Serine dehydratase kn-keyword=Serine dehydratase en-keyword=d-Amino acid kn-keyword=d-Amino acid END start-ver=1.4 cd-journal=joma no-vol=15 cd-vols= no-issue=19 article-no= start-page=6336 end-page=6352 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20071001 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Antitumor studies. Part 3: Design, synthesis, antitumor activity, and molecular docking study of novel 2-methylthio-, 2-amino-, and 2-(N-substituted amino)-10-alkyl-2-deoxo-5-deazaflavins en-subtitle= kn-subtitle= en-abstract= kn-abstract=Various novel 10-alkyl-2-deoxo-2-methylthio-5-deazaflavins have been synthesized by reaction of 6-(N-alkylanilino)-2-methylthiopyrimidin-4(3H)-ones with Vilsmeier reagent. The similar 2-(N-substituted amino) derivatives were prepared by nucleophilic replacement reaction of the 2-methylthio moiety by appropriate amines. The 2-oxo derivatives (i.e., 5-deazaflavins) were obtained by acidic hydrolysis of the 2-methylthio derivatives. The antitumor activities against CCRF-HSB-2 and KB cells and the antiviral activities against HSV-1 and HSV-2 have been investigated in vitro, and many compounds showed promising antitumor activities. Furthermore, AutoDock molecular docking into PTK has been done for lead optimization of these compounds as potential PTK inhibitors. Whereas, the designed 2-deoxo-5-deazaflavins connected with amino acids at the 2-position exhibited the good binding affinities into PTK with more hydrogen bonds. en-copyright= kn-copyright= en-aut-name=AliHamed I. en-aut-sei=Ali en-aut-mei=Hamed I. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=AshidaNoriyuki en-aut-sei=Ashida en-aut-mei=Noriyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NagamatsuTomohisa en-aut-sei=Nagamatsu en-aut-mei=Tomohisa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= affil-num=1 en-affil= kn-affil=Department of Drug Discovery and Development, Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University affil-num=2 en-affil= kn-affil=Biology Laboratory, Research and Development Division, Yamasa Shoyu Co. affil-num=3 en-affil= kn-affil=aDepartment of Drug Discovery and Development, Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University en-keyword=antitumor activity kn-keyword=antitumor activity en-keyword=5-deazaflavin kn-keyword=5-deazaflavin en-keyword=AutoDock kn-keyword=AutoDock en-keyword=protein tyrosine kn-keyword=protein tyrosine en-keyword=kinase kn-keyword=kinase END start-ver=1.4 cd-journal=joma no-vol=73 cd-vols= no-issue=4 article-no= start-page=281 end-page=285 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20070705 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=N-terminal domain including conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Flagellin in Pseudomonas syringae is a potent elicitor of defense responses including hypersensitive cell death in dicot plants. The oligopeptides flg22 consisting of 22 conserved amino acids near the N-terminus of flagellins is reported to induce plant defense responses. Because glycosylation of the central domain of flagellin affects its elicitor activity, we investigated whether any peptide sequence in addition to flg22 is required for flagellin-induced hypersensitive reaction. A study of recombinant flagellin polypeptides indicated that the N-terminal domain including the conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana.

en-copyright= kn-copyright= en-aut-name=KanaNaito en-aut-sei=Kana en-aut-mei=Naito kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=YasuhiroIshiga en-aut-sei=Yasuhiro en-aut-mei=Ishiga kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KazuhiroToyoda en-aut-sei=Kazuhiro en-aut-mei=Toyoda kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=TomonoriShiraishi en-aut-sei=Tomonori en-aut-mei=Shiraishi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YukiIchinose en-aut-sei=Yuki en-aut-mei=Ichinose kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Graduate School of Natural Science and Technology, Okayama University affil-num=2 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Graduate School of Natural Science and Technology, Okayama University affil-num=3 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Graduate School of Natural Science and Technology, Okayama University affil-num=4 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Graduate School of Natural Science and Technology, Okayama University affil-num=5 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Graduate School of Natural Science and Technology, Okayama University en-keyword=Arabidopsis thaliana kn-keyword=Arabidopsis thaliana en-keyword=Flagellin kn-keyword=Flagellin en-keyword=Flg22 kn-keyword=Flg22 en-keyword=FLS2 kn-keyword=FLS2 en-keyword=HR cell death kn-keyword=HR cell death END start-ver=1.4 cd-journal=joma no-vol=189 cd-vols= no-issue=19 article-no= start-page=6945 end-page=6956 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=200710 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Flagellin Glycans from two pathovars of Pseudomonas syringae contain rhamnose in D and L configurations in different ratios and modified 4-amino-4,6-dideoxyglucose en-subtitle= kn-subtitle= en-abstract= kn-abstract=Flagellins from Pseudomonas syringae pv. glycinea race 4 and Pseudomonas syringae pv. tabaci 6605 have been found to be glycosylated. Glycosylation of flagellin is essential for bacterial virulence and is also involved in the determination of host specificity. Flagellin glycans from both pathovars were characterized, and common sites of glycosylation were identified on six serine residues (positions 143, 164, 176, 183, 193, and 201). The structure of the glycan at serine 201 (S201) of flagellin from each pathovar was determined by sugar composition analysis, mass spectrometry, and H-1 and C-13 nuclear magnetic resonance spectroscopy. These analyses showed that the S201 glycans from both pathovars were composed of a common unique trisaccharide consisting of two rhamnosyl (Rha) residues and one modified 4-amino-4,6-dideoxyglucosyl (Qui4N) residue, beta-D-Quip4N(3-hydroxy-1-oxobutyl)2Me-(1 -> 3)-alpha-L-Rhap-(1 -> 2)-alpha-L-Rhap. Furthermore, mass analysis suggests that the glycans on each of the six serine residues are composed of similar trisaccharide units. Determination of the enantiomeric ratio of Rha from the flagellin proteins showed that flagellin from P. syringae pv. tabaci 6605 consisted solely Of L-Rha, whereas P. syringae pv. glycinea race 4 flagellin contained both L-Rha and D-Rha at a molar ratio of about 4:1. Taking these findings together with those from our previous study, we conclude that these flagellin glycan structures may be important for the virulence and host specificity of P. syringae. en-copyright= kn-copyright= en-aut-name=TakeuchiKasumi en-aut-sei=Takeuchi en-aut-mei=Kasumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OnoHiroshi en-aut-sei=Ono en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YoshidaMitsuru en-aut-sei=Yoshida en-aut-mei=Mitsuru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=IshiiTadashi en-aut-sei=Ishii en-aut-mei=Tadashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=KatohEtsuko en-aut-sei=Katoh en-aut-mei=Etsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TaguchiFumiko en-aut-sei=Taguchi en-aut-mei=Fumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MikiRyuji en-aut-sei=Miki en-aut-mei=Ryuji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=MurataKatsuyoshi en-aut-sei=Murata en-aut-mei=Katsuyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KakuHanae en-aut-sei=Kaku en-aut-mei=Hanae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= affil-num=1 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=2 en-affil= kn-affil=National Food Research Institute affil-num=3 en-affil= kn-affil=National Food Research Institute affil-num=4 en-affil= kn-affil=Forestry and Forest Products Research Institute affil-num=5 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=6 en-affil= kn-affil=Graduate School of Natural Science and Technology, Okayama University affil-num=7 en-affil= kn-affil=Graduate School of Natural Science and Technology, Okayama University affil-num=8 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=9 en-affil= kn-affil=Faculty of Agriculture, Meiji University affil-num=10 en-affil= kn-affil=Graduate School of Natural Science and Technology, Okayama University en-keyword=INNATE IMMUNE-RESPONSE kn-keyword=INNATE IMMUNE-RESPONSE en-keyword=TOLL-LIKE RECEPTOR-5 kn-keyword=TOLL-LIKE RECEPTOR-5 en-keyword=PV. TABACI kn-keyword=PV. TABACI en-keyword=POSTTRANSLATIONAL MODIFICATION kn-keyword=POSTTRANSLATIONAL MODIFICATION en-keyword=BACTERIAL FLAGELLIN kn-keyword=BACTERIAL FLAGELLIN en-keyword=STRUCTURAL-ANALYSIS kn-keyword=STRUCTURAL-ANALYSIS en-keyword=AMINO-ACIDS kn-keyword=AMINO-ACIDS en-keyword=GLYCOSYLATION kn-keyword=GLYCOSYLATION en-keyword=AERUGINOSA kn-keyword=AERUGINOSA en-keyword=IDENTIFICATION kn-keyword=IDENTIFICATION END start-ver=1.4 cd-journal=joma no-vol=73 cd-vols= no-issue=5 article-no= start-page=831 end-page=837 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20070522 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Functionalization of chitosan with 3-nitro-4-amino benzoic acid moiety and its application to the collection/concentration of molybdenum in environmental water samples en-subtitle= kn-subtitle= en-abstract= kn-abstract=

A chitosan resin functionalized with 3-nitro-4-amino benzoic acid moiety (CCTS-NABA resin) was newly synthesized for the collection/concentration of trace molybdenum by using cross-linked chitosan (CCTS) as base material. The carboxyl group of the moiety was chemically attached to amino group of cross-linked chitosan through amide bond formation. The adsorption behavior of molybdenum as well as other 60 elements on the resin was examined by passing the sample solutions through a mini-column packed with the resin. After the elution of the elements collected on the resin with 1 M HNO3, the eluates were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) and atomic emission spectrometry (ICP-AES).
The CCTS-NABA resin can adsorb several metal ions, such as vanadium, gallium, arsenic, selenium, silver, bismuth, thorium, tungsten, tin, tellurium, copper, and molybdenum at appropriate pHs. Among these metal ions, only molybdenum could be adsorbed almost completely on the resin at acidic regions. An excellent selectivity toward molybdenum could be obtained at pH 3-4. The adsorption capacity of CCTS-NABA resin for Mo(VI) was 380 mg g(-1) resin. Through the column pretreatment, alkali and alkaline earth metals in river water and seawater samples were successfully removed.
The CCTS-NABA resin was applied to the adsorption/collection of molybdenum in river water and seawater samples. The concentrations of molybdenum in river water samples were found in the range of 0.84 and 0.95 ppb (ng g(-1)), whereas molybdenum in seawater was about 9 ppb. The validation of the proposed method was carried out by determining molybdenum in the certified reference materials of SLRS-4, CASS-4, and NASS-5 after passing through the CCTS-NABA resin; the results showed good agreement with the certified-values.

en-copyright= kn-copyright= en-aut-name=SabarudinAkhmad en-aut-sei=Sabarudin en-aut-mei=Akhmad kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OshimaMitsuko en-aut-sei=Oshima en-aut-mei=Mitsuko kn-aut-name=‘哇ŒυŽq kn-aut-sei=‘哇 kn-aut-mei=ŒυŽq aut-affil-num=2 ORCID= en-aut-name=NoguchiOsamu en-aut-sei=Noguchi en-aut-mei=Osamu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MotomizuShoji en-aut-sei=Motomizu en-aut-mei=Shoji kn-aut-name=–{…Ή“ρ kn-aut-sei=–{… kn-aut-mei=Ή“ρ aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Brawijaya University affil-num=2 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University en-keyword=chitosan resin kn-keyword=chitosan resin en-keyword=3-nitro-4-amino benzoic acid kn-keyword=3-nitro-4-amino benzoic acid en-keyword=molybdenum kn-keyword=molybdenum en-keyword=adsorption behavior kn-keyword=adsorption behavior en-keyword=water and seawater kn-keyword=water and seawater en-keyword=ICP-MS kn-keyword=ICP-MS en-keyword=ICP-AES kn-keyword=ICP-AES en-keyword=certified reference materials kn-keyword=certified reference materials END start-ver=1.4 cd-journal=joma no-vol=72 cd-vols= no-issue=5 article-no= start-page=1609 end-page=1617 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20070118 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Sequential-injection on-line preconcentration using chitosan resin functionalized with 2-amino-5-hydroxy benzoic acid for the determination of trace elements in environmental water samples by inductively coupled plasma-atomic emission spectrometry en-subtitle= kn-subtitle= en-abstract= kn-abstract=

A new chelating resin using chitosan as a base material was synthesized. Functional moiety of 2-amino-5-hydroxy benzoic acid (AHBA) was chemically bonded to the amino group of cross-linked chitosan (CCTS) through the arm of chloromethyloxirane (CCTS-AHBA resin). Several elements, such as Ag, Be, Cd, Co, Cu, Ni, Ph, U, V, and rare earth elements (REEs), could be adsorbed on the resin. To use the resin for on-line pretreatment, the resin was packed in a mini-column and installed into a sequential-injection/automated pretreatment system (Auto-Pret System) coupled with inductively coupled plasma-atomic emission spectrometry (ICP-AES). The sequential-injection/automated pretreatment system was a laboratory-assembled, and the program was written using Visual Basic software. This system can provide easy operation procedures, less reagent consumption, as well as less waste production. Experimental variables considered as effective factors in the improvement sensitivity, such as an eluent concentration, a sample and an eluent flow rate, pH of samples, and air-sandwiched eluent were carefully optimized. The proposed system provides excellent on-line collection efficiency, as well as high concentration factors of analytes in water samples, which results in highly sensitive detection of ultra-trace and trace analysis. Under the optimal conditions, the detection limits of 24 elements examined are in the range from ppt to sub-ppb levels. The proposed method was validated by using the standard reference material of a river water, SLRS-4, and the applicability was further demonstrated to the on-line collection/concentration of trace elements, such as Ag, Be, Cd, Co, Cu, Ni, Ph, U, V, and REEs in water samples.

en-copyright= kn-copyright= en-aut-name=SabarudinAkhmad en-aut-sei=Sabarudin en-aut-mei=Akhmad kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=LenghorNarong en-aut-sei=Lenghor en-aut-mei=Narong kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OshimaMitsuko en-aut-sei=Oshima en-aut-mei=Mitsuko kn-aut-name=‘哇ŒυŽq kn-aut-sei=‘哇 kn-aut-mei=ŒυŽq aut-affil-num=3 ORCID= en-aut-name=HakimLukman en-aut-sei=Hakim en-aut-mei=Lukman kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TakayanagiToshio en-aut-sei=Takayanagi en-aut-mei=Toshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=GaoYun-Hua en-aut-sei=Gao en-aut-mei=Yun-Hua kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=MotomizuShoji en-aut-sei=Motomizu en-aut-mei=Shoji kn-aut-name=–{…Ή“ρ kn-aut-sei=–{… kn-aut-mei=Ή“ρ aut-affil-num=7 ORCID= affil-num=1 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=2 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=5 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=6 en-affil= kn-affil=Technical Institute of Physics and Chemistry, Chinese Academy of Sciences (CAS) affil-num=7 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University en-keyword=sequential-injection kn-keyword=sequential-injection en-keyword=on-line preconcentration kn-keyword=on-line preconcentration en-keyword=trace elements kn-keyword=trace elements en-keyword=ICP-AES kn-keyword=ICP-AES en-keyword=chitosan resin kn-keyword=chitosan resin END start-ver=1.4 cd-journal=joma no-vol=23 cd-vols= no-issue=12 article-no= start-page=1431 end-page=1434 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=20071211 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Adsorption behavior of cationic and anionic species on chitosan resins possessing amino acid moieties en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Chitosan resins modified with amino acids, such as glycine, valine, leucine, and serine, were synthesized for investigating the adsorption behavior of cationic and anionic species, and showed good abilities for the adsorption of trace elements in aquatic media as follows: glycine for lanthanoids at pH 7, leucine for molybdenum at pH 1 - 5, serine for uranium at pH 2 - 7, and amino acids for bismuth at pH 1 - 7. Cationic and anionic species could be adsorbed by a chelating mechanism and an anion-exchange mechanism.

en-copyright= kn-copyright= en-aut-name=OshitaKoji en-aut-sei=Oshita en-aut-mei=Koji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TakayanagiToshio en-aut-sei=Takayanagi en-aut-mei=Toshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OshimaMitsuko en-aut-sei=Oshima en-aut-mei=Mitsuko kn-aut-name=‘哇ŒυŽq kn-aut-sei=‘哇 kn-aut-mei=ŒυŽq aut-affil-num=3 ORCID= en-aut-name=MotomizuShoji en-aut-sei=Motomizu en-aut-mei=Shoji kn-aut-name=–{…Ή“ρ kn-aut-sei=–{… kn-aut-mei=Ή“ρ aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of International Conservation Studies for Cultural Properties, Faculty of Cultural Properties, Kibi International University affil-num=2 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University en-keyword=CHEMICALLY-MODIFIED CHITOSAN kn-keyword=CHEMICALLY-MODIFIED CHITOSAN en-keyword=SELECTIVE ADSORPTION kn-keyword=SELECTIVE ADSORPTION en-keyword=ICP-MS kn-keyword=ICP-MS en-keyword=COLUMN COLLECTION/CONCENTRATION kn-keyword=COLUMN COLLECTION/CONCENTRATION en-keyword=COMPLEXANE TYPES kn-keyword=COMPLEXANE TYPES en-keyword=PRECIOUS METALS kn-keyword=PRECIOUS METALS en-keyword=DERIVATIVES kn-keyword=DERIVATIVES en-keyword=ION kn-keyword=ION en-keyword=COPRECIPITATION kn-keyword=COPRECIPITATION en-keyword=COPPER(II) kn-keyword=COPPER(II) END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=1 article-no= start-page=136 end-page=144 dt-received= dt-revised= dt-accepted= dt-pub-year=2004 dt-pub=20041208 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Synthesis of cross-linked chitosan possessing N-methyl-D-glucamine moiety (CCTS-NMDG) for adsorption/concentration of boron in water samples and its accurate measurement by ICP-MS and ICP-AES en-subtitle= kn-subtitle= en-abstract= kn-abstract=

A chitosan resin derivatized with N-methyl-(D)-glucamine (CCTS-NMDG) was synthesized by using a cross-linked chitosan (CCTS) as base material. The N-methyl-D-glucamine (NMDG) moiety was attached to the amino group of CCTS through the arm of chloromethyloxirane. The adsorption behavior of 59 elements on the synthesized resin was systematically examined by using the resin packed in a mini-column, passing water samples through it and measuring the adsorbed elements in eluates by ICP-MS. The CCTS-NMDG resin shows high ability in boron sorption with the capacity of 0.61 mmol ml(-1) (= 2.1 mmol g(-1)). The sorption kinetics of this resin was faster than that of the commercially available resins. Other advantages of the synthesized resin are: (1) quantitative collection of boron at neutral pH regions; (2) complete removal of large amounts of matrices; (3) no loss of efficiency over prolonged usage; (4) effective collection of boron in wide range concentration using a mini column containing 1 ml resin; (5) complete elution of boron with 1 mol 1(-1) nitric acid. The resin was applied to the collection/concentration of boron in water samples. Boron in tap water and river water was found to be in the range of 6-8 mu g 1(-1). The limit of detection (LOD) of boron after pretreatment with CCTS-NMDG resin and measurement by ICP-MS was 0.07 mu g 1(-1) and the limit of quantification (LOQ) was 0. 14 mu g 1(-1) when the volume of each sample and eluent was 10 ml.

en-copyright= kn-copyright= en-aut-name=SabarudinAkhmad en-aut-sei=Sabarudin en-aut-mei=Akhmad kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OshitaKoji en-aut-sei=Oshita en-aut-mei=Koji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OshimaMitsuko en-aut-sei=Oshima en-aut-mei=Mitsuko kn-aut-name=‘哇ŒυŽq kn-aut-sei=‘哇 kn-aut-mei=ŒυŽq aut-affil-num=3 ORCID= en-aut-name=MotomizuShoji en-aut-sei=Motomizu en-aut-mei=Shoji kn-aut-name=–{…Ή“ρ kn-aut-sei=–{… kn-aut-mei=Ή“ρ aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=2 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Chemistry, Faculty of Science, Okayama University en-keyword=chitosan resin kn-keyword=chitosan resin en-keyword=N-methyl-D-glucamine kn-keyword=N-methyl-D-glucamine en-keyword=boron kn-keyword=boron en-keyword=adsorption kn-keyword=adsorption en-keyword=ICP-MS/AES kn-keyword=ICP-MS/AES END start-ver=1.4 cd-journal=joma no-vol=125 cd-vols= no-issue=16 article-no= start-page=3133 end-page=3141 dt-received= dt-revised= dt-accepted= dt-pub-year=1998 dt-pub=19988 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The ich1 gene of the mushroom Coprinus cinereus is essential for pileus formation in fruiting en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The formation of the pileus in homobasidiomycete fungi is essential for sexual reproduction, because the pileus bears the hymenium, a layer of cells that includes the specialised basidia in which nuclear fusion, meiosis and sporulation occur. The developmental mutant ichijiku of Coprinus cinereus fails to develop a differentiated pileus at the apex of the primordial shaft, which is the basal part of the fruit-body primordia and formed in an early stage of fruit-body differentiation. Genetic analysis indicates that this phenotype is caused by a recessive mutation in a single gene (ich1). The ich1 gene was mapped to chromosome XII using restriction fragment length polymorphism markers and the marker chromosome method, and cloned by complementation using a chromosome-XII-specific cosmid library. The ich1 gene encodes a novel protein of 1,353 amino acids. The Ich1 amino-acid sequence contains nuclear targeting signals, suggesting that the Ich1 protein would function in the nucleus. Northern blot analysis indicates that the ich1 gene is specifically expressed in the pileus of the wild-type fruit-body. No ich1 mRNA was detected in the ichijiku mutant, consistent with loss of the promoter region of ich1 in the mutant genome. These data demonstrate that the ick1 gene product is essential for pileus formation.

en-copyright= kn-copyright= en-aut-name=MuraguchiHajime en-aut-sei=Muraguchi en-aut-mei=Hajime kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KamadaTakashi en-aut-sei=Kamada en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University en-keyword=Developmental mutant kn-keyword=Developmental mutant en-keyword=Ich1 kn-keyword=Ich1 en-keyword=Pileus kn-keyword=Pileus en-keyword=Fruit-body kn-keyword=Fruit-body en-keyword=morphogenesis kn-keyword=morphogenesis en-keyword=Basidiomycete kn-keyword=Basidiomycete en-keyword=Coprinus kn-keyword=Coprinus END start-ver=1.4 cd-journal=joma no-vol=271 cd-vols= no-issue=1 article-no= start-page=1 end-page=10 dt-received= dt-revised= dt-accepted= dt-pub-year=2004 dt-pub=20041 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Structure and expression of 12-oxophytodienoate reductase (OPR) subgroup I gene in pea and oxidoreductase activity of their recombinant proteins en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Recently, we observed that expression of a pea gene (S64) encoding an oxophytodienoic acid reductase (OPR) was induced by a suppressor of pea defense responses, secreted by the pea pathogen Mycosphaerella pinodes. Because it is known that OPRs are usually encoded by families of homologous genes, we screened for genomic and cDNA clones encoding members of this putative OPR family in pea. We isolated five members of the OPR gene family from a pea genomic DNA library, and amplified six cDNA clones, including S64, by RT-PCR (reverse transcriptase-PCR). Sequencing analysis revealed that S64 corresponds to PsOPR2, and the amino acid sequences of the predicted products of the six OPR-like genes shared more than 80% identity with each other. Based on their sequence similarity, all these OPR-like genes code for OPRs of subgroup I, i.e., enzymes which are not required for jasmonic acid biosynthesis. However, the genes varied in their exon/intron organization and in their promoter sequences. To investigate the expression of each individual OPR-like gene, RT-PCR was performed using gene-specific primers. The results indicated that the OPR-like gene most strongly induced by the inoculation of pea plants with a compatible pathogen and by treatment with the suppressor from M. pinodes was PsOPR2. Furthermore, the ability of the six recombinant OPR-like proteins to reduce a model substrate, 2-cyclohexen-1-one (2-CyHE), was investigated. The results indicated that PsOPR1, 4 and 6 display robust activity, and PsOPR2 has a most remarkable ability to reduce 2-CyHE, whereas PsOPR3 has little and PsOPR5 does not reduce this compound. Thus, the six OPR-like proteins can be classified into four types. Interestingly, the gene structures, expression profiles, and enzymatic activities used to classify each member of the pea OPR-like gene family are clearly correlated, indicating that each member of this OPR-like family has a distinct function.

en-copyright= kn-copyright= en-aut-name=MatsuiH en-aut-sei=Matsui en-aut-mei=H kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NakamuraG en-aut-sei=Nakamura en-aut-mei=G kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IshigaY en-aut-sei=Ishiga en-aut-mei=Y kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=ToshimaH en-aut-sei=Toshima en-aut-mei=H kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=InagakiY en-aut-sei=Inagaki en-aut-mei=Y kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=ToyodaK en-aut-sei=Toyoda en-aut-mei=K kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=ShiraishiT en-aut-sei=Shiraishi en-aut-mei=T kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=2 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=3 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=4 en-affil= kn-affil=Ibaraki University affil-num=5 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=6 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=7 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University affil-num=8 en-affil= kn-affil=Laboratory of Plant Pathology and Genetic Engineering, Faculty of Agriculture, Okayama University en-keyword=coronatine kn-keyword=coronatine en-keyword=flavoproteins kn-keyword=flavoproteins en-keyword=Jasmonic acid kn-keyword=Jasmonic acid en-keyword=oxophytodienoic acid reductase kn-keyword=oxophytodienoic acid reductase en-keyword=OPR kn-keyword=OPR en-keyword=suppressor kn-keyword=suppressor END start-ver=1.4 cd-journal=joma no-vol=207 cd-vols= no-issue=4 article-no= start-page=621 end-page=632 dt-received= dt-revised= dt-accepted= dt-pub-year=2004 dt-pub=20042 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Purification and cDNA cloning of the ovigerous-hair stripping substance (OHSS) contained in the hatch water of an estuarine crab Sesarma haematocheir en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The egg attachment system of an estuarine crab Sesarma haematocheir is formed on the maternal ovigerous hairs just after egg laying, and slips off these hairs just after hatching. The stripping is caused by an active factor that we call OHSS (ovigerous-hair stripping substance), which is released by the embryo upon hatching. OHSS was purified, and its active form had a molecular mass of 25?kDa. The cDNA of OHSS cloned from an embryonic cDNA library was 1759?bp long, encoding 492 amino acids in a single open reading frame (ORF). The C-terminal part of the predicted protein was composed of a trypsin-like serine protease domain, with homology to counterparts in other animals of 33?38%. The predicted protein (54.7?kDa) secreted as a zymogen may be cleaved post-translationally, separating the Cterminal from the N-terminal region. The OHSS gene was expressed in the embryo at least 2 weeks before hatching. Expression was also detected in the zoea larva 1 day after hatching and in the brain of the female. However, it was not detected in the muscle, hepatopancreas or ovigerous seta of the female. Ultrastructural analysis indicated that the material investing maternal ovigerous hair, i.e. the outermost layer (E1) of the egg case, is attached at the special sites (attachment sites) arranged at intervals of 130?160?nm on the hair. It is suggested that OHSS acts specifically at these sites, lysing the bond with the coat, thus disposing of the embryo attachment system. This enables the female to prepare the next clutch of embryos without ecdysis.

en-copyright= kn-copyright= en-aut-name=GusevOleg en-aut-sei=Gusev en-aut-mei=Oleg kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IkedaHideki en-aut-sei=Ikeda en-aut-mei=Hideki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OkochiTetsushi en-aut-sei=Okochi en-aut-mei=Tetsushi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=LeeJae Min en-aut-sei=Lee en-aut-mei=Jae Min kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=HatakeyamaMasatsugu en-aut-sei=Hatakeyama en-aut-mei=Masatsugu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KobayashiChiyoko en-aut-sei=Kobayashi en-aut-mei=Chiyoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=AgataKiyokazu en-aut-sei=Agata en-aut-mei=Kiyokazu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=YamadaHidenori en-aut-sei=Yamada en-aut-mei=Hidenori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=SaigusaMasayuki en-aut-sei=Saigusa en-aut-mei=Masayuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama Univeristy affil-num=4 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=5 en-affil= kn-affil=National Institute of Agrobiological Sciences, Owashi affil-num=6 en-affil= kn-affil=RIKEN, Hyougo affil-num=7 en-affil= kn-affil=RIKEN, Hyougo affil-num=8 en-affil= kn-affil=Okayama Univresity affil-num=9 en-affil= kn-affil=Okayama University en-keyword=crab kn-keyword=crab en-keyword=Sesarma (or Chiromantes) haematocheir kn-keyword=Sesarma (or Chiromantes) haematocheir en-keyword=ovigerous hair kn-keyword=ovigerous hair en-keyword=embryo attachment system kn-keyword=embryo attachment system en-keyword=investment coat kn-keyword=investment coat en-keyword=stripping kn-keyword=stripping en-keyword= ovigerous-hair stripping substance (OHSS) kn-keyword= ovigerous-hair stripping substance (OHSS) en-keyword= serine protease. kn-keyword= serine protease. END start-ver=1.4 cd-journal=joma no-vol=8 cd-vols= no-issue=6 article-no= start-page=923 end-page=938 dt-received= dt-revised= dt-accepted= dt-pub-year=2006 dt-pub=20066 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Identification of glycosylation genes and glycosylated amino acids of flagellin in Pseudomonas syringae pv. tabaci en-subtitle= kn-subtitle= en-abstract= kn-abstract=A glycosylation island is a genetic region required for glycosylation. The glycosylation island of flagellin in Pseudomonas syringae pv. tabaci 6605 consists of three orfs: orf1, orf2 and orf3. Orf1 and orf2 encode putative glycosyltransferases, and their deletion mutants, Delta orf1 and Delta orf2, exhibit deficient flagellin glycosylation or produce partially glycosylated flagellin respectively. Digestion of glycosylated flagellin from wild-type bacteria and non-glycosylated flagellin from Delta orf1 mutant using aspartic N-peptidase and subsequent HPLC analysis revealed candidate glycosylated amino acids. By generation of site-directed Ser/Ala-substituted mutants, all glycosylated amino acid residues were identified at positions 143, 164, 176, 183, 193 and 201. Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) analysis revealed that each glycan was about 540 Da. While all glycosylation-defective mutants retained swimming ability, swarming ability was reduced in the Delta orf1, Delta orf2 and Ser/Ala-substituted mutants. All glycosylation mutants were also found to be impaired in the ability to adhere to a polystyrene surface and in the ability to cause disease in tobacco. Based on the predicted tertiary structure of flagellin, S176 and S183 are expected to be located on most external surface of the flagellum. Thus the effect of Ala-substitution of these serines is stronger than that of other serines. These results suggest that glycosylation of flagellin in P. syringae pv. tabaci 6605 is required for bacterial virulence. It is also possible that glycosylation of flagellin may mask elicitor function of flagellin molecule. en-copyright= kn-copyright= en-aut-name=TaguchiFumiko en-aut-sei=Taguchi en-aut-mei=Fumiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TakeuchiKasumi en-aut-sei=Takeuchi en-aut-mei=Kasumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KatohEtsuko en-aut-sei=Katoh en-aut-mei=Etsuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MurataKatsuyoshi en-aut-sei=Murata en-aut-mei=Katsuyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SuzukiTomoko en-aut-sei=Suzuki en-aut-mei=Tomoko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=MarutaniMizuri en-aut-sei=Marutani en-aut-mei=Mizuri kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=KawasakiTakayuki en-aut-sei=Kawasaki en-aut-mei=Takayuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=EguchiMinako en-aut-sei=Eguchi en-aut-mei=Minako kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=KatohShizue en-aut-sei=Katoh en-aut-mei=Shizue kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=kakuHanae en-aut-sei=kaku en-aut-mei=Hanae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=YasudaChihiro en-aut-sei=Yasuda en-aut-mei=Chihiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=InagakiYoshishige en-aut-sei=Inagaki en-aut-mei=Yoshishige kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= en-aut-name=ToyodaKazuhiro en-aut-sei=Toyoda en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=13 ORCID= en-aut-name=ShiraishiTomonori en-aut-sei=Shiraishi en-aut-mei=Tomonori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=14 ORCID= en-aut-name=IchinoseYuki en-aut-sei=Ichinose en-aut-mei=Yuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=15 ORCID= affil-num=1 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=2 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=3 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=4 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=5 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=6 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=7 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=8 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=9 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=10 en-affil= kn-affil=National Institute of Agrobiological Sciences affil-num=11 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=12 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=13 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=14 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University affil-num=15 en-affil= kn-affil=The Graduate School of Natural Science and Technology, Okayama University en-keyword=Gram-Negative bacteria kn-keyword=Gram-Negative bacteria en-keyword=Posttranslational modification kn-keyword=Posttranslational modification en-keyword=Protein Glycosylation kn-keyword=Protein Glycosylation en-keyword=Perception kn-keyword=Perception en-keyword=Aeruginosa kn-keyword=Aeruginosa en-keyword=Cells kn-keyword=Cells en-keyword=Specificity kn-keyword=Specificity en-keyword=Expression kn-keyword=Expression en-keyword=Plasmids kn-keyword=Plasmids en-keyword=Pathways kn-keyword=Pathways END start-ver=1.4 cd-journal=joma no-vol=162 cd-vols= no-issue=1 article-no= start-page=73 end-page=79 dt-received= dt-revised= dt-accepted= dt-pub-year=2004 dt-pub=20042 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Separation into polar and hydrogen-bonding factors of the effects of alcohols on the emission spectrum of 4-phenyl-1-N,N-dimethylaminobutane in THF en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The effects of the additions of protic and aprotic polar solvents on the emission spectrum of 4-phenyl-1-N,N-dimethylaminobutane (PDAB) in THF have been studied under conditions of steady-state illumination. The fluorescence spectrum of PDAB in THF was reported to consist of three component bands (band A at 285 nm (fluorescence of the phenyl group), band B at 343 nm (fluorescence of the amino group) and band C at 385 nm (emission from an intramolecular exciplex)). The intensities of bands B and C decreased with increasing solvent polarity. They also decreased owing to the hydrogen-bonding interaction between the amino group in PDAB and protic solvents, but in this case the intensity of band A was found to increase. Acetonitrile has only a polar effect and trichloroacetic acid only a hydrogen-bonding (or protonation) effect, while alcohols have both effects. The equilibrium constants for the formation of intermolecular hydrogen-bonded complexes of the amino group with alcohols were estimated from the intensity change of band A. The hydrogen-bonding and polar effects of alcohols on the intensities of bands B and C could be separately evaluated. The decrease in the intensities of bands B and C with increasing solvent polarity in THF-AN and THF-alcohol mixtures is considered to be caused by the conversion of the exciplex to an ion-pair enhanced by the increase in solvent polarity.

en-copyright= kn-copyright= en-aut-name=GuobinXie en-aut-sei=Guobin en-aut-mei=Xie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=NakamuraMayuko en-aut-sei=Nakamura en-aut-mei=Mayuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=SueishiYoshimi en-aut-sei=Sueishi en-aut-mei=Yoshimi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamamotoShunzo en-aut-sei=Yamamoto en-aut-mei=Shunzo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama Univresity affil-num=4 en-affil= kn-affil=Okayama University en-keyword=solvent effect kn-keyword=solvent effect en-keyword=polar effect kn-keyword=polar effect en-keyword=hydrogen-bonding kn-keyword=hydrogen-bonding en-keyword=fluorescence kn-keyword=fluorescence en-keyword=exciplex kn-keyword=exciplex END start-ver=1.4 cd-journal=joma no-vol=64 cd-vols= no-issue=1 article-no= start-page=49 end-page=54 dt-received= dt-revised= dt-accepted= dt-pub-year=2010 dt-pub=201002 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The First Case of a Class I Glucose-6-phosphate Dehydrogenase Deficiency, G6PD Santiago de Cuba (1339 GA), in a Chinese Population as Found in a Survey for G6PD Deficiency in Northeastern and Central China en-subtitle= kn-subtitle= en-abstract= kn-abstract=

In Liaoning Province in northeastern China, we found a G6PD-deficient patient at the age of 3. By the classification of the World Health Organization, this patient was categorized as class I (very severe G6PD deficiency). When we investigated the G6PD gene of the patient, we found that he had a replacement of G to A at nucleotide 1339. As a result, the amino acid at position 447 should change from Gly to Arg. This replacement is known as G6PD Santiago de Cuba, because it was first discovered in a Cuban boy who showed heavy chronic anemia. Today, 28 G6PD variants have been reported in the Chinese population, and all are categorized as class II (severe deficiency) or class III (mild deficiency);in class II or III deficiency, anemia is not present in daily life, but hemolytic attack can occur when the carrier ingests certain oxidative medicines or foods. This is the first report of a G6PD-deficient Chinese patient in the category of class I. We intended to find other G6PD-deficient cases in northeastern China and tested several hundred blood samples, but no cases of G6PD deficiency were found (0/414). In central China, where falciparum malaria was endemic from the 1950s to 1970s, we found two G6PD-deficient cases (2/27) and the other members from their families whose variant type was G6PD Kaiping (1388GT), which is a common variant in the Chinese population.

en-copyright= kn-copyright= en-aut-name=WangJichun en-aut-sei=Wang en-aut-mei=Jichun kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MatsuokaHiroyuki en-aut-sei=Matsuoka en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=HiraiMakoto en-aut-sei=Hirai en-aut-mei=Makoto kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MuLing en-aut-sei=Mu en-aut-mei=Ling kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YangLiandi en-aut-sei=Yang en-aut-mei=Liandi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=LuoEnjie en-aut-sei=Luo en-aut-mei=Enjie kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=Department of Pathogen Biology, China Medical University affil-num=2 en-affil= kn-affil=Division of Medical Zoology, Department of Infection and Immunity, Jichi Medical University affil-num=3 en-affil= kn-affil=Division of Medical Zoology, Department of Infection and Immunity, Jichi Medical University affil-num=4 en-affil= kn-affil=Shenyang Infectious Disease Hospital affil-num=5 en-affil= kn-affil=Hubei Center for Disease Control and Prevention affil-num=6 en-affil= kn-affil=Department of Pathogen Biology, China Medical University en-keyword=hemolytic anemia kn-keyword=hemolytic anemia en-keyword=Chinese kn-keyword=Chinese en-keyword=glucose-6-phosphate dehydrogenase kn-keyword=glucose-6-phosphate dehydrogenase en-keyword=G6PD Santiago de Cuba kn-keyword=G6PD Santiago de Cuba en-keyword=malaria kn-keyword=malaria END start-ver=1.4 cd-journal=joma no-vol=57 cd-vols= no-issue=3 article-no= start-page=117 end-page=122 dt-received= dt-revised= dt-accepted= dt-pub-year=2003 dt-pub=200306 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effect of ornithine on the ileal histology, nitric oxide production and lipid peroxidation in LPS-induced endotoxemia. en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Effect of ornithine which is known to inhibit L-arginine uptake via cationic amino acid transport system has been tested, and compared to aminoguanidine, an iNOS inhibitor in lypopolysaccharide (LPS)-induced endotoxemia in rats. Serum nitrite/nitrate and malondialdehyde (MDA) level have been measured, and ileal histology has also been examined. Endotoxin increased serum nitrite/nitrate and MDA levels from 15.7+/- 2.4 micromol/ml and 2.1 +/-0.2 nmol/ml to 23.1 +/- 1.0 micromol/ml and 5.2+/- 0.3 nmol/ml (both P<0.05), respectively. In addition, LPS caused ileal degeneration. L-ornithine (500 mg/kg) did not improve septic manifestations, i.e., serum nitrite/nitrate and MDA levels did not differ from those in endotoxemia. Neither does it have an improving action on ileal histology. However, higher dose of L-ornithine (2,500 mg/kg) lowered the increased level of nitrite/nitrate and MDA by LPS. Moreover, it restored ileal histology from grade 3 (median) to 0 (median) (P<0.05). On the other hand, aminoguanidine (100 mg/kg) normalized serum nitrite/nitrate and MDA levels but not ileal histology in endotoxemic rats. In conclusion, high dose of L-ornithine could improve endotoxemic parameters in LPS-treated rats.

en-copyright= kn-copyright= en-aut-name=DirlikMusa en-aut-sei=Dirlik en-aut-mei=Musa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=BuyukafsarKansu en-aut-sei=Buyukafsar en-aut-mei=Kansu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=CinelIsmail en-aut-sei=Cinel en-aut-mei=Ismail kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=CinelLeyla en-aut-sei=Cinel en-aut-mei=Leyla kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=CaglikulekciMehmet en-aut-sei=Caglikulekci en-aut-mei=Mehmet kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TamerLulufer en-aut-sei=Tamer en-aut-mei=Lulufer kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=AydinSuha en-aut-sei=Aydin en-aut-mei=Suha kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=OralUgur en-aut-sei=Oral en-aut-mei=Ugur kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Mersin University affil-num=2 en-affil= kn-affil=Mersin University affil-num=3 en-affil= kn-affil=Mersin University affil-num=4 en-affil= kn-affil=Mersin University affil-num=5 en-affil= kn-affil=Mersin University affil-num=6 en-affil= kn-affil=Mersin University affil-num=7 en-affil= kn-affil=Mersin University affil-num=8 en-affil= kn-affil=Mersin University en-keyword=LPS kn-keyword=LPS en-keyword=ornithine kn-keyword=ornithine en-keyword=aminoguanidine kn-keyword=aminoguanidine en-keyword=endotoxemia kn-keyword=endotoxemia en-keyword=lipid peroxidation kn-keyword=lipid peroxidation END start-ver=1.4 cd-journal=joma no-vol=24 cd-vols= no-issue=3 article-no= start-page=333 end-page=342 dt-received= dt-revised= dt-accepted= dt-pub-year=1970 dt-pub=197006 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Glutamic acid metabolism in perfused cat brain studied with 14C-labelled glutamic acid en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The rate of transport of blood glutamic acid into the brain and the rate of metabolic conversion of the amino acid in the brain were derived by the use of the brain perfution method in vivo and in situ with [D.HC] ?Lglutamic acid. The net uptake of glutamic acid by the brain was observed. Most of the radioactivity released from the brain into the cerebral venous blood was found to consist of glutamine. Small but significant amounts of output as radioactive GSH and CO2 were also found. Glutamic acid transport and its rate of metabolism were lowered in the glucose. free condition. The size of the compartment of the small glutamic acid pool, which was related closely to the blood glutamic acid, and that of the large glutamic acid pool, which was related closely to the blood glucose, were calculated and compared with each other.

en-copyright= kn-copyright= en-aut-name=WatanabeShosuke en-aut-sei=Watanabe en-aut-mei=Shosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OtsukiSaburo en-aut-sei=Otsuki en-aut-mei=Saburo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NakashimaYoshio en-aut-sei=Nakashima en-aut-mei=Yoshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=EdamatsuKazuyasu en-aut-sei=Edamatsu en-aut-mei=Kazuyasu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=MitsunobuKatsusuke en-aut-sei=Mitsunobu en-aut-mei=Katsusuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama University affil-num=4 en-affil= kn-affil=Okayama University affil-num=5 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=28 cd-vols= no-issue=4 article-no= start-page=243 end-page=246 dt-received= dt-revised= dt-accepted= dt-pub-year=1974 dt-pub=197408 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Effect of pyridoxine treatment of a homocystinuric patient on the urinary excretion of some sulfur-containing amino acids en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The effect of pyridoxine treatment of a homocystinuric patient on the urinary excretion of some sulfur-containing amino acids was studied and the following results were obtained. As a result of pyridoxine treatment, urinary homocystine decreased to a fairly great extent, and its unusual metabolites S.(3-hydroxy-3-carboxyn- propylthio) homocysteine (HCPTHC) and S-C8-carboxyethylthio homocysteine (j3-CETHC) increased to some extent. But its oxidation product (homocysteic acid) showed a tendency to decrease slightly. Urinary methionine and cystine increased to some extent, but cysteinehomocysteine mixed disulfide showed no remarkable change.

en-copyright= kn-copyright= en-aut-name=KodamaH. en-aut-sei=Kodama en-aut-mei=H. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=IkegamiT. en-aut-sei=Ikegami en-aut-mei=T. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=YaoK. en-aut-sei=Yao en-aut-mei=K. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=OuraT. en-aut-sei=Oura en-aut-mei=T. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama University affil-num=4 en-affil= kn-affil=Children's Medical Center oj Osaka City END start-ver=1.4 cd-journal=joma no-vol=46 cd-vols= no-issue=3 article-no= start-page=169 end-page=174 dt-received= dt-revised= dt-accepted= dt-pub-year=1992 dt-pub=199206 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Concentration and uptake of taurine in umbilical blood platelets. en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The concentration and uptake of taurine in the umbilical and adult blood platelets were studied. Taurine was the most abundant free amino acid in both umbilical and adult blood platelets. The taurine concentration in umbilical blood platelets (2.30 pmoles/10(4) cells) was significantly lower than that of adult blood platelets (3.27 pmoles/10(4) cells) in contrast to the reverse relationship in taurine concentrations in umbilical and adult blood plasma. No other amino acid showed such significant difference in the concentrations between umbilical and adult blood platelets. Taurine uptake into umbilical blood platelets was temperature sensitive and sodium-dependent in a manner similar to that of adult blood platelets. The uptake conformed well to Hanes-plot. The Vmax of the uptake into adult blood platelets was about 3.6 times higher than that of umbilical blood platelets, but no significant difference was seen in the Km value between the two groups.

en-copyright= kn-copyright= en-aut-name=KanemoriHirofumi en-aut-sei=Kanemori en-aut-mei=Hirofumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=EjiriKohei en-aut-sei=Ejiri en-aut-mei=Kohei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AkahoriShuichiro en-aut-sei=Akahori en-aut-mei=Shuichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KuboTakafumi en-aut-sei=Kubo en-aut-mei=Takafumi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SekibaKaoru en-aut-sei=Sekiba en-aut-mei=Kaoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama Univerisity affil-num=4 en-affil= kn-affil=Okayama University affil-num=5 en-affil= kn-affil=Okayama University en-keyword=umbilical blood kn-keyword=umbilical blood en-keyword=platelet kn-keyword=platelet en-keyword=taurine concentration kn-keyword=taurine concentration en-keyword=faurine uptake kn-keyword=faurine uptake END start-ver=1.4 cd-journal=joma no-vol=46 cd-vols= no-issue=5 article-no= start-page=337 end-page=343 dt-received= dt-revised= dt-accepted= dt-pub-year=1992 dt-pub=199210 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=NG-nitro-L-arginine attenuates flow debt repayment in the reactive hyperemic response of the open-chest dog coronary artery: contribution of endothelium-derived relaxing factor. en-subtitle= kn-subtitle= en-abstract= kn-abstract=

To test the hypothesis that the endothelium-derived relaxing factor (EDRF) contributes to coronary vasodilation induced by myocardial ischemia, we examined the effect of NG-nitro-L-arginine (a potent and selective inhibitor of EDRF release) on the coronary reactive hyperemic response in the open-chest dogs. Intracoronary infusion of NG-nitro-L-arginine at a coronary plasma concentration of 5 x 10(-5) M had no effect on hemodynamics and myocardial oxygen metabolism, but attenuated repayment of the flow debt by an average of 20.4% and 20.0% following coronary occlusion for 10 sec and 20 sec, respectively. Concomitant infusion of NG-nitro-L-arginine at the same concentration and 8-phenyltheophylline (a potent adenosine receptor blocker) at a coronary plasma concentration of 10(-5) M further attenuated flow debt repayment following 10 sec and 20 sec of coronary occlusion by 47.7 and 59.4%, respectively. These results indicate that EDRF plays a significant role in the coronary reactive hyperemic response and may cause vasodilation independently of adenosine-mediated vasodilation following coronary occlusion.

en-copyright= kn-copyright= en-aut-name=MarutaniMorio en-aut-sei=Marutani en-aut-mei=Morio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KusachiShouzo en-aut-sei=Kusachi en-aut-mei=Shouzo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KajikawaYutaka en-aut-sei=Kajikawa en-aut-mei=Yutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=YamasakiSatoshi en-aut-sei=Yamasaki en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TsujiTakao en-aut-sei=Tsuji en-aut-mei=Takao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama University affil-num=4 en-affil= kn-affil=Okayama University affil-num=5 en-affil= kn-affil=Okayama University en-keyword=myocardial reactive hyperemia kn-keyword=myocardial reactive hyperemia en-keyword=nitric oxide kn-keyword=nitric oxide en-keyword=amino acids kn-keyword=amino acids en-keyword=metabolic vasodilation kn-keyword=metabolic vasodilation END start-ver=1.4 cd-journal=joma no-vol=46 cd-vols= no-issue=6 article-no= start-page=401 end-page=405 dt-received= dt-revised= dt-accepted= dt-pub-year=1992 dt-pub=199212 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Sulfur Amino Acid Levels and Related Enzyme Activities in Various Brain Regions (and Other Tissues) in Normal Mice and Rolling Mice Nagoya en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The contents of the sulfur amino acids, and the activities of cystathionine beta-synthase and cystathionine gamma-lyase were measured in various regions of the brain and several other tissues in both normal mice and rolling mice Nagoya. The cystathionine content and cystathionine beta-synthase activity were found to be unevenly distributed in various regions of the brain in both normal mice and rolling mice Nagoya, being highest in the cerebellum. Except for the mesencephalon and thalamus plus hypothalamus, the cystathionine content and cystathionine beta-synthase activity in the brain regions of rolling mice Nagoya were much higher than those of the normal mice. The cystathionine content after D,L-propargylglycine treatment was also found to be unevenly distributed in various brain regions in both normal mice and rolling mice Nagoya. The concentrations of cystine and methionine were also higher in all regions of the brain of rolling mice Nagoya than those of normal mice, while the concentration of taurine in the various regions of the brain was almost the same in normal mice and rolling mice Nagoya. Cystathionine beta-synthase and cystathionine gamma-lyase activities in the liver, kidney, and pancreas were almost the same in both the normal mice and rolling mice Nagoya.

en-copyright= kn-copyright= en-aut-name=MyojinKazuhiro en-aut-sei=Myojin en-aut-mei=Kazuhiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HiroiTadashi en-aut-sei=Hiroi en-aut-mei=Tadashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=IkedaHisao en-aut-sei=Ikeda en-aut-mei=Hisao kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KodamaHiroyuki en-aut-sei=Kodama en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Kochi Medical School affil-num=2 en-affil= kn-affil=Kochi Medical School affil-num=3 en-affil= kn-affil=Kochi Medical School affil-num=4 en-affil= kn-affil=Kochi Medical School en-keyword=cystathionine kn-keyword=cystathionine en-keyword=rolling mouse Nagoya kn-keyword=rolling mouse Nagoya en-keyword=brain kn-keyword=brain en-keyword=propargylglycine kn-keyword=propargylglycine END start-ver=1.4 cd-journal=joma no-vol=17 cd-vols= no-issue=6 article-no= start-page=273 end-page=278 dt-received= dt-revised= dt-accepted= dt-pub-year=1963 dt-pub=1963 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Experimental isovalthinuria. III. Induction by bile acids, and hypocholesterolemic agents en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Some bile acids (dehydrocholic, cholic, chenodeoxycholic, ursodeoxycholic, and deoxycholic acids), and some hypocholesterolemic agents (22, 25 diazacholestanol, 20,25-diazacholesterol, triparanol, and SKF 525-A) are the inducers of isovalthinuria in guinea pig. Administration of methionine appears to increase the pool of sulfur compound which participates in the formation of isovalthine. Cholesterol appears to have no enhancing effect on the induction activity of isovalthinuria inducers. The mechanism of isovalthine formation and the role of sulfur amino acids in lowering blood cholesterol are discussed.

en-copyright= kn-copyright= en-aut-name=UbukaToshihiko en-aut-sei=Ubuka en-aut-mei=Toshihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=22 cd-vols= no-issue=2 article-no= start-page=101 end-page=112 dt-received= dt-revised= dt-accepted= dt-pub-year=1968 dt-pub=196804 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Nucleic acids and protein synthesis in cancer cell mitochondria. II. Amino acid incorporation into proteins of rat liver and hepatoma cell mitochondria en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The energy source required for the amino acid incorporation into mitochondrial proteins has been investigated and comparative study has also been made on the rate of the amino acid incorporation in rat liver and rat hepatoma cell mitochondria. 1. The incorporation of amino acid into the protein in intact mitochondria of rat liver increased by about 40% on the addition of α-ketoglutarate and ADP, but no significant increase in the amino acid incorporation was observed on the addition of succinate and ADP. 2. The incorporation of amino acids into mitochondrial proteins was remarkably inhibited by the addition of respiratory inhibitors (cyanide, DNP at a high concentration). 3. The amino acid incorporation into mitochondrial proteins was scarcely or slightly inhibited by the addition of DNP at the concentration of 1~10-4M and insensitive to oligomycin (5 to 10 μg/ml). 4. The amino acid incorporation into the protein in the endogenous substrate system of the mitochondria was considerably inhibited by the addition of arsenite, and this inhibition somewhat recovered on the addition of ADP plus succinate. 5. The rate of the amino acid incorporations between rat liver and hepatoma cell mitochondria was at the same level. 6. Discussions were made on the energy source required for the amino acid incorporation into mitochondrial proteins, on the rate of protein synthesis per mitochondrion isolated from rat liver- and hepatoma cells, and on the possibilities of contamination of bacteria or microsomes and of the adsorption of amino acids onto the mitochondria.

en-copyright= kn-copyright= en-aut-name=InabaKozo en-aut-sei=Inaba en-aut-mei=Kozo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=23 cd-vols= no-issue=6 article-no= start-page=497 end-page=503 dt-received= dt-revised= dt-accepted= dt-pub-year=1969 dt-pub=196912 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Experimental isovalthinuria. VII. Experiments with radioactive acetate, valine, and leucine en-subtitle= kn-subtitle= en-abstract= kn-abstract=

1. For the settlement of carbon origin of urinary isovalthine, acetic acid-2-C14, valine-U-C14 or leucine-U-C14 was administered to rats together with isovaleric acid as an isovalthinuria inducer, and urinary isovalthine excreted was tested by autoradiography. As the results of which, it was found that these isotopic compounds were not the precursor of urinary isovalthine. Although the isovalthinuria inducing effect of isovaleric acid was fairly diminished by these isotopic compounds, urinary isovalthine was detected by paper electrophoresis. 2. Some metabolic products of these isotopic compounds were also inquired in urine and some tissues. The results were as follows: a) Acetic acid incorporated into urea, aspartate, serine, glutamate, proline, glycine, alanine, ornithine, ethanolamine, r-amino-buthyric acid (brain only), cholesterol and fatty acids. b) Valine incorporated into urinary glutamate and glycine, and tissue cholesterol and fatty acids. Valine was rapidly excreted in urine and found in a very small amount in liver digest. c) Leucine incorporated into urinary aspartate, serine, glutamate and glycine, and tissue cholesterol and fatty acids. 3. Several important problems of isovalthine studies to be elucidated were discussed.

en-copyright= kn-copyright= en-aut-name=FjiiYoshio en-aut-sei=Fjii en-aut-mei=Yoshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=21 cd-vols= no-issue=6 article-no= start-page=321 end-page=326 dt-received= dt-revised= dt-accepted= dt-pub-year=1967 dt-pub=196712 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Studies on nitrogen pool of animal tissues. 3. Ox liver and bile. IV. Ox kidney and lung en-subtitle= kn-subtitle= en-abstract= kn-abstract=

The compositions of nitrogen pools of ox liver, bladder bile, kidney and lung were analyzed with an especial bearing on their minor components, and some distinctive features of these tissues were described. DCEC and CMC were found in ox liver and kidney. Liver was low in free arginine and lysine, but high in ornithine, ethanolamine, and glutathione. Glycine was only a predominant amino acid in ox bile. All amino acids were contained moderately in kidney, but glutathione content was low. The concentrations of arginine and lysine were relatively high in lung.

en-copyright= kn-copyright= en-aut-name=AzumiTsukasa en-aut-sei=Azumi en-aut-mei=Tsukasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=21 cd-vols= no-issue=6 article-no= start-page=315 end-page=320 dt-received= dt-revised= dt-accepted= dt-pub-year=1967 dt-pub=196712 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Studies on nitrogen pool of animal tissues. I. Ox ocular tissues. II. Ox nervous tissues en-subtitle= kn-subtitle= en-abstract= kn-abstract=

Concentrations of ampholytes in the nitrogen pool of ox ocular tissues and nervous tissues were analyzed systematically by an automatic amino acid analyzer with a special reference to their minor components. DCEC was found in lens and also in nervous tissues. Ophthalmic acid was found in lens (highest), in retina (moderate), and in vitreous humor and spinal cord (trace). Glutathione content was extremely high in lens, and moderate in nervous tissues, retina and cornea. Carnosine content was moderate in cornea and in retina, but hemocarnosine may be rather high in nervous tissues. Anserine-like compound was found only in spinal cord, but free 1- and 3-methylhistidine were detected in most ocular tissues. Ethanolamine and γ-aminobutyric acid were high in retina and their concentrations were comparable to those of nervous tissues.

en-copyright= kn-copyright= en-aut-name=AzumiTsukasa en-aut-sei=Azumi en-aut-mei=Tsukasa kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= affil-num=1 en-affil= kn-affil=Okayama University END start-ver=1.4 cd-journal=joma no-vol=21 cd-vols= no-issue=6 article-no= start-page=279 end-page=296 dt-received= dt-revised= dt-accepted= dt-pub-year=1967 dt-pub=196712 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Regulatory effects of blood constituents on the function and metabolism of the cat brain in perfusion ezperiments. Brain perfusion with artificial blood containing low molecular dextran and amino acids en-subtitle= kn-subtitle= en-abstract= kn-abstract=

As a link in a series of studies on the effects of blood constituents on the brain function by means of brain perfusion, we used four kinds of artificial blood; namely, the blood containing a low molecular dextran, one containing glutamic acid, one containing essential amino acid group and the one containing both essential amino acid group and glutamic acid. During the perfusion experiments we observed the effects of blood constituents on the function and metabolism of the perfused brain and obtained the following results. 1. When a low molecular dextran is used as the colloid osmotic pressure agent instead of hydrodextran, the amount of the blood flow in the brain is maintained roughly at a certain fixed level throughout the experiment, showing no gradual decreasing tendency. 2. When using the artificial blood supplemented with glutamic acid, EEG of the perfused brain shows an increase in the appearance rate of β32 and β33 bands, approaching closely to the pattern of EEG of unrestrained controls at arousal state. 3. In the case of the blood added with essential amino acids similar to the case using the blood with glutamic acid, EEG approaches towards the alert pattern of the controls. 4. When the perfusion is done with the artificial blood lacking in amino acids, about one hour after the start of the perfusion the amount of glutamic acid and its related compounds in the brain can no longer be maintained at normal level and the decrease, being so marked, brings about a marked decrease also in total amino acid content. 5. When the perfusion blood contains glutamic acid, essential amino acid group or both, the concentrations of amino acids of the brain glutamic acid group and the total amino acid can be maintained approximately at normal level for the duration of over one hour.

en-copyright= kn-copyright= en-aut-name=OtsukiSaburo en-aut-sei=Otsuki en-aut-mei=Saburo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=WatanabeShosuke en-aut-sei=Watanabe en-aut-mei=Shosuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MorimitsuJunsuke en-aut-sei=Morimitsu en-aut-mei=Junsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=EdamatsuKazuyasu en-aut-sei=Edamatsu en-aut-mei=Kazuyasu kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=NakashimaYoshihiko en-aut-sei=Nakashima en-aut-mei=Yoshihiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=OkumuraNikichi en-aut-sei=Okumura en-aut-mei=Nikichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=Okayama University affil-num=2 en-affil= kn-affil=Okayama University affil-num=3 en-affil= kn-affil=Okayama University affil-num=4 en-affil= kn-affil=Okayama University affil-num=5 en-affil= kn-affil=Okayama University affil-num=6 en-affil= kn-affil=Okayama University END