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Journal of Biomedical Science and Engineering 2022年第2期15卷 83-88页

作者： tadashi miura Yukari Oda Yasuhiko Shizawa Oral Health Science Center Tokyo Dental College Tokyo Japan Department of Oral and Maxillofacial Implantology Tokyo Dental College Tokyo Japan Department of Liberal Arts College of Bioresource Sciences Nihon University Kanagawa Japan

Newly synthesized membrane and secretory proteins in cells undergo folding in the endoplasmic reticulum with the introduction of disulfide bonds and acquire the correct three-dimensional structure. Disulfide bonds are especially important for protein folding. It has been thought that formation of protein disulfide bonds in eukaryotes is mainly carried out by an enzyme called protein disulfide isomerase. Proteins, bearing the C-terminus of amino acids sequences with His-Asp-Glu-Leu (HDEL) sequence in yeast, in the endoplasmic reticulum (ER), which is a eukaryotic cellular organelle involved in protein synthesis, processing, and transport, have been considered to recycle between ER and Golgi apparatus. The proposal for this recycling model derives from the study of an HDEL-tagged fusion protein. Here, the localization and oligosaccharide modification of protein disulfide isomerase were investigated in yeast, and showed the first direct evidence that this intrinsic ER protein transports from ER to Golgi. Results suggest that this native protein is also accessible to post-ER enzymes, and yet accumulates in the ER.

关键词： Protein Disulfide Isomerase Endoplasmic Reticulum Golgi Apparatus Oligosaccharide Modification Protein Localization

Ca (II)-EDTA shows antimicrobial activity against periodontopathic bacteria

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Journal of Biomedical Science and Engineering 2012年第1期5卷 10-14页

作者： tadashi miura Koji Tanabe Masao Yoshinari 不详

Ethylenediamine-N,N,N’,N’-tetraacetic acid calcium (II) disodium salt (Ca (II)-EDTA) was investigated for its antibacterial activity against the periodontal pathogens Aggregatibacter actinomycetemcomitans, Prevotella intermedia and Porphyromonas gingivalis. We determined the minimum inhibitory concentrations (MIC) of Ca (II)-EDTA and its bactericidal activity. Ethylenediamine-N,N,N’,N’-tetraacetic acid calcium (II) disodium salt inhibited the growth of all the periodontopathic bacteria tested in broth. An MIC of 60 mM was the most effective against P. gingivalis. An adenosine triphosphate bioluminescence assay revealed that Ca (II)-EDTA showed bactericidal activity against the bacteria tested in a time-dependent manner. To determine its safety in mammalian cells, we investigated the viability of murine L929 cells subjected to Ca (II)-EDTA treatment. At 75 mM, 93% ± 0.13% of the cells survived. These results indicate that Ca (II)-EDTA is a candidate chelating agent for prevention of infection by periodontopathic bacteria.

关键词： Chelating Agent Periodontopathic Bacteria Peri-Implantitis Antimicrobial Activity

Effect of zeta potentials on bovine serum albumin adsorption on crown composite resin surfaces <i>in vitro</i>

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Journal of Biomedical Science and Engineering 2013年第3期6卷 273-276页

作者： Nahoko Miyake tadashi miura Toru Sato Masao Yoshinari Department of Crown & Bridge Prosthodontics Tokyo Dental College Chiba Japan Division of Oral Implants Research Oral Health Science Center Tokyo Dental College Chiba Japan Division of Prosthodontics Department of Clinical Oral Health Science Tokyo Dental College Tokyo Japan

We previously studied the mechanism underlying the adsorption of oral bacteria on the surfaces of dental prosthetic materials such as ceramics and resins in vitro. The aim of the present study was to examine bovine serum albumin (BSA) adsorption on crown composite resin surfaces by means of zeta potential. We measured the zeta potentials of resins alone, BSA alone, and resins after BSA adsorption. Eight resins were pulverized into powders (300 - 1000 nm). All experiments were conducted in 10 mM sodium chloride solution (pH 6.5). BSA was dissolved in 10 mM NaCl with a concentration of 2.0 × 10-5 mol/l. An adsorption assay was performed for one hour at 37°C under continuous rotation (6 rpm). The zeta potentials of both resins and BSA were negative, with BSA itself less negative than the resins themselves as an absolute value (p < 0.0001). The zeta potentials of seven resin surfaces after BSA adsorption were significantly less negative than were those of the resins without BSA adsorption (p < 0.0001). Eight resins were divided into two classes based on the size of the surface potential difference between each resin and the BSA. The difference in surface potential between the resins and the BSA were small, leading to the theory that particles with identical charges repulse each other, and the amounts of adsorbed BSA on these resins might be less. On the other, when the differences between the other resins and BSA are large, so that the repulsive force between two nonidentical particles becomes zero and an attractive force might be generated, then more BSA might be adsorbed on those resins. Therefore, the zeta potentials were affected by BSA adsorption and became less negative. These results suggested that electrostatic interactions play an important role in the adsorption of BSA on resin surfaces.

关键词： Zeta Potential Crown Composite Resins Bovine Serum Albumin Adsorption Electrostatic Interactions

Basic peptide protamine exerts antimicrobial activity against periodontopathic bacteria——Growth inhibition of periodontopathic bacteria by protamine

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Journal of Biomedical Science and Engineering 2010年第11期3卷 1069-1072页

作者： tadashi miura Keishi Iohara Tetsuo Kato Kazuyuki Ishihara Masao Yoshinari 不详

Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibitory concentrations of protamine and its hydrolysate and their bactericidal activity. Protamine inhibited the growth of all periodontopathic bacteria tested on agar plates. Protamine, which MIC was 6.3 × 10-7 g L-1, was most effective against P. gingivalis. The antibacterial effect of native protamine was higher than that of its hydrolysate. An ATP bioluminescence assay revealed that protamine showed bactericidal activity against P. gingivalis in a time-dependent manner. These results indicate that protamine could be candidate peptide for prevention of P. gingivalis infection.

关键词： Antimicrobial Peptide Periodontopathic Bacteria Implantitis Protamine

Antimicrobial Effects of Gatifloxacin on Canine Periodontopathic Bacteria In Vitro

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Journal of Biomedical Science and Engineering 2022年第2期15卷 89-96页

作者： tadashi miura Yukari Oda Koji Tanabe Yasuhiko Shizawa Taichi Ito Longqiang Yang Oral Health Science Center Tokyo Dental College Tokyo Japan Department of Oral and Maxillofacial Implantology Tokyo Dental College Tokyo Japan Department of Biomedical Engineering Iwate Medical University Iwate Japan Department of Liberal Arts College of Bioresource Sciences Nihon University Kanagawa Japan Department of Pediatric Dentistry Sichuan Hospital of Stomatology Chengdu China

The aim of this study was to identify the potential antibacterial effects of gatifloxacin on one of canine periodontal pathogens, Porphyromonas gulae. The minimum inhibitory concentrations (MIC) of gatifloxacin and its bactericidal effects were investigated. Gatifloxacin inhibited the growth of the canine periodontopathic bacteria tested in broth. An MIC of 50 nM was found to be effective in inhibiting P. gulae. On performing adenosine triphosphate bioluminescence assay, gatifloxacin was found to exhibit bactericidal effects on the tested bacteria in a concentration-dependent manner. The safety of gatifloxacin in mammalian cells was evaluated by assessing the viability of rat bone marrow mesenchymal stem (BMMS) cells treated with gatifloxacin. Almost over 80% of BMMS cells survived after a 3-day culture when treated with 100 nM of gatifloxacin. These results indicate that locally administered gatifloxacin has the potential for being used to prevent canine periodontal infection.

关键词： Periodontal Disease Antibiotics Dog Porphyromonas gulae Veterinary Medicine

Debridement Effect on Periodontal Pathogen <i>Porphyromonas gingivalis</i>Cultured on Titanium by Application of Atmospheric-Pressure Plasma

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Journal of Biomedical Science and Engineering 2017年第2期10卷 51-59页

作者： tadashi miura Masahiro Egawa Taichi Ito Toru Eguro Koji Tanabe Masao Yoshinari Oral Health Science Center Tokyo Dental College Tokyo Japan Department of Periodontology Tokyo Dental College Chiba Japan Department of Oral and Maxillofacial Implantology Tokyo Dental College Chiba Japan Japan Institute for Advanced Dentistry Tokyo Japan Department of Pharmacology Tokyo Dental College Tokyo Japan

The debridement treatments of dental implants are very important in long-term maintenance after implant placement in a patient. Deposition of periodontal pathogens around the implant surface has a high risk of causing periimplantitis. The aim of this study was to evaluate the extent of elimination of Porphyromonas gingivalis, known as representative periodontopathic bacteria, from titanium, which has been the main material used for dental implants. Assuming that the debridement processing of dental implants removes periodontal bacteria, one of the methods for removing bacteria deposited on titanium is considered to be plasma irradiation. Irradiation with atmospheric-pressure plasma was carried out against periodontopathic bacteria cultured and deposited on the surface of a titanium disk. After the plasma irradiation, the reduction of the number of bacteria re-cultured for 24 hours was evaluated. The number of viable bacteria on the titanium surface was estimated by an ATP-bioluminescent assay. Viable cells after the plasma irradiation were reduced to 1.5% or less compared to the untreated group. As one of the methods of debridement in general dental treatments, atmospheric-pressure plasma has proved to be an effective method to remove adverse prognostic factors in dental patients.

关键词： Porphyromonas gingivalis Atmospheric-Pressure Plasma Debridement Peri-Implantitis Titanium

Potential Antimicrobial Effects of Gatifloxacin on Periodontopathic Bacteria in Vitro

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Journal of Biomedical Science and Engineering 2016年第7期9卷 354-359页

作者： tadashi miura Koji Tanabe Eri Tsukagoshi Koichi Kida Yasuhiko Shizawa Nahoko Miyake Masataka Kasahara Masao Yoshinari Division of Oral Implants Research Oral Health Science Center Tokyo Dental College Tokyo Japan Department of Pharmacology Tokyo Dental College Tokyo Japan Department of Oral and Maxillofacial Implantology Tokyo Dental College Tokyo Japan Department of Liberal Arts College of Bioresource Sciences Nihon University Kanagawa Japan Department of Removable Partial Prosthodontics Tokyo Dental College Tokyo Japan

The aim of this study was to identify the potential antibacterial effects of gatifloxacin on periodontal pathogens including Aggregatibacter actinomycetemcomitans, Porphyromonas gingi-valis, and Prevotella intermedia. The minimum inhibitory concentrations (MIC) of gatifloxacin and its bactericidal effects were investigated. Gatifloxacin inhibited the growth of all three kinds of periodontopathic bacteria tested in broth. The MIC value of 2.5 nM was found to be the most effective in inhibiting A. actinomycetemcomitans. An adenosine triphosphate biolumi-nescence assay revealed that gatifloxacin exhibited bactericidal effects on the tested bacteria in a time-dependent manner. The safety of gatifloxacin in mammalian cells was evaluated by assessing the viability of normal human dermal fibroblast (NHDF) cells treated with gatifloxacin. Almost all NHDF cells survived after 2-d culture, while 81% of the cells survived after 4-d culture when treated with 1.0 × 103 nM gatifloxacin. These results indicate that gatifloxacin is a possible drug for local administration to prevent periodontal infection.

关键词： Local Administration Peri-Implantitis Periodontopathic Bacteria Antimicrobial Activity Gatifloxacin

PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells(HDDPCs) and HDDPC-derived iPS cells

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International Journal of Oral Science 2015年第3期7卷 144-154页

作者： Emi Inada Issei Saitoh Satoshi Watanabe Reiji Aoki Hiromi miura Masato Ohtsuka Tomoya Murakami tadashi Sawami Youichi Yamasaki Masahiro Sato Department of Pediatric Dentistry Kagoshima University Graduate School of Medical and Dental Sciences Division of Pediatric Dentistry Graduate School of Medical and Dental Science Niigata University Animal Genome Research Unit Division of Animal Science National Institute of Agrobiological Sciences Functional Biomolecules Research Group National Agriculture and Food Research Organization Division of Basic Molecular Science and Molecular Medicine School of Medicine Tokai University Section of Gene Expression Regulation Frontier Science Research Center Kagoshima University

The ability of human deciduous tooth dental pulp cells（HDDPCs） to differentiate into odontoblasts that generate mineralized tissue holds immense potential for therapeutic use in the field of tooth regenerative medicine. Realization of this potential depends on efficient and optimized protocols for the genetic manipulation of HDDPCs. In this study, we demonstrate the use of a Piggy Bac（PB）-based gene transfer system as a method for introducing nonviral transposon DNA into HDDPCs and HDDPC-derived inducible pluripotent stem cells. The transfection efficiency of the PB-based system was significantly greater than previously reported for electroporation-based transfection of plasmid DNA. Using the neomycin resistance gene as a selection marker, HDDPCs were stably transfected at a rate nearly 40-fold higher than that achieved using conventional methods. Using this system, it was also possible to introduce two constructs simultaneously into a single cell. The resulting stable transfectants, expressing td Tomato and enhanced green fluorescent protein, exhibited both red and green fluorescence. The established cell line did not lose the acquired phenotype over three months of culture. Based on our results, we concluded that PB is superior to currently available methods for introducing plasmid DNA into HDDPCs. There may be significant challenges in the direct clinical application of this method for human dental tissue engineering due to safety risks and ethical concerns. However, the high level of transfection achieved with PB may have significant advantages in basic scientific research for dental tissue engineering applications, such as functional studies of genes and proteins. Furthermore, it is a useful tool for the isolation of genetically engineered HDDPC-derived stem cells for studies in tooth regenerative medicine.

关键词： drug selection electroporation genetically modified human deciduous tooth dental pulp cells Piggy Bac

Non-cooperative optimization games in market-oriented overlay networks： an integrated model of resource pricing and network formation

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Frontiers of Computer Science 2011年第4期5卷 496-505页

作者： Yutaka OKAIE tadashi NAKANO Graduate School of Engineering Osaka University Suita Osaka 565-0871 Japan

In this paper, we formulate a non-cooperative optimization game in market-oriented overlay networks where participating peers share their own computing resources to earn virtual money called energy. We model an overlay network as a set of non-cooperative resource providing peers, called platforms, that perform resource pricing and topology management to maximize their own energy gains. Resource consuming peers, called agents, are simply designed to migrate platform-to-platform to find the least expensive resources in the network. Simulation results are presented to demonstrate the market dynamics as well as the global properties of the network, i.e., resource price and network topology, that emerge from local interactions among the group of peers.

关键词： Overlay networks market mechanisms non-cooperative games mobile agents

Amelanotic malignant melanoma of the esophagus:Report of two cases with immunohistochemical and molecular genetic study of KIT and PDGFRA

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World Journal of Gastroenterology 2009年第21期15卷 2679-2683页

作者： tadashi Terada Department of Pathology Shizuoka City Shimizu Hospital

The author reports herein two cases of amelanotic malignant melanoma of the esophagus. Case 1 is an 87-year-old woman who was admitted to our hospital because of nausea and vomiting. Endoscopic examination revealed an ulcerated tumor of the distal esophagus, and a biopsy was taken. The biopsy showed malignant polygonal and spindle cells. No melanin pigment was recognized. Immunohistochemically, the tumor cells were positive for melanosome （HMB45）, S100 protein, KIT and Platelet derived growth factor receptor-α （PDG- FRA）. The patient was treated by chemotherapy and radiation, but died of systemic metastasis 12 mo after the presentation. Case 2 is a 56-year-old man presenting with dysphagia. Endoscopic examination revealed a polypoid tumor in the middle esophagus, and a biopsy was obtained. The biopsy showed malignant spindle cells without melanin pigment. Immunohistochemically, the tumor cells were positively labeled for melanosome,S100 protein, KIT and PDGFRA. The patient refused operation, and was treated by palliative chemotherapy and radiation. He died of metastasis 7 mo aEer the admission. In both cases, molecular genetic analyses of gene （exons 9, 11, 13 and 17） and PDGFRA gene （exons 12 and 18） were performed by the PCR direct sequencing method, which showed no mutations of KIT and PDGFRA genes. This is the first report of esophageal malignant melanoma with an examination of the expression of KIT and PDGFRA and the mutational status of K/T and PDGFRA genes.

关键词： Esophagus Amelanotic melanoma KIT Platelet derived growth factor receptor-c~