Elsevier Science Acta Medica Okayama 0969-8051 43 6 2016 Noninvasive evaluation of nicotinic acetylcholine receptor availability in mouse brain using single-photon emission computed tomography with [(123)I]5IA. 372 378 EN Yuki Matsuura Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Masashi Ueda Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Yusuke Higaki Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Keiko Watanabe Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Shogo Habara Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Shinichiro Kamino Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University Hideo Saji Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto University Shuichi Enomoto Department of Pharmaceutical Analytical Chemistry, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University INTRODUCTION: Nicotinic acetylcholine receptors (nAChRs) are of great interest because they are implicated in higher brain functions. Nuclear medical imaging is one of the useful techniques for noninvasive evaluation of physiological and pathological function in living subjects. Recent progress in nuclear medical imaging modalities enables the clear visualization of the organs of small rodents. Thus, translational research using nuclear medical imaging in transgenic mice has become possible and helps to elucidate human disease pathology. However, imaging of α4β2 nAChRs in the mouse brain has not yet been performed. The purpose of this study was to assess the feasibility of single-photon emission computed tomography (SPECT) with 5-[(123)I]iodo-3-[2(S)-azetidinylmethoxy]pyridine ([(123)I]5IA) for evaluating α4β2 nAChR availability in the mouse brain. METHODS: A 60-min dynamic SPECT imaging session of α4β2 nAChRs in the mouse brain was performed. The regional distribution of radioactivity in the SPECT images was compared to the density of α4β2 nAChRs measured in an identical mouse. Alteration of nAChR density in the brains of Tg2576 mice was also evaluated. RESULTS: The mouse brain was clearly visualized by [(123)I]5IA-SPECT and probe accumulation was significantly inhibited by pretreatment with (-)-nicotine. The regional distribution of radioactivity in SPECT images showed a significant positive correlation with α4β2 nAChR density measured in an identical mouse brain. Moreover, [(123)I]5IA-SPECT was able to detect the up-regulation of α4β2 nAChRs in the brains of Tg2576 transgenic mice. CONCLUSIONS: [(123)I]5IA-SPECT imaging would be a promising tool for evaluating α4β2 nAChR availability in the mouse brain and may be useful in translational research focused on nAChR-related diseases. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 0304-4165 1863 4 2019 Regulation of Ca2+/calmodulin-dependent protein kinase kinase beta by cAMP signaling 672 680 EN Shota Takabatake Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Satomi Ohtsuka Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Takeyuki Sugawara Department of Anatomy, Kitasato University School of Medicine Naoya Hatano Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Naoki Kanayama Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Masaki Magari Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Hiroyuki Sakagami Department of Anatomy, Kitasato University School of Medicine Hiroshi Tokumitsu Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University BACKGROUND: Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) is a pivotal activator of CaMKI, CaMKIV and 5'-AMP-activated protein kinase (AMPK), controlling Ca2+-dependent intracellular signaling including various neuronal, metabolic and pathophysiological responses. Recently, we demonstrated that CaMKKβ is feedback phosphorylated at Thr144 by the downstream AMPK, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme. However, the regulatory phosphorylation of CaMKKβ at Thr144 in intact cells and in vivo remains unclear. METHODS: Anti-phosphoThr144 antibody was used to characterize the site-specific phosphorylation of CaMKKβ in immunoprecipitated samples from mouse cerebellum and in transfected mammalian cells that were treated with various agonists and protein kinase inhibitors. CaMKK activity assay and LC-MS/MS analysis were used for biochemical characterization of phosphorylated CaMKKβ. RESULTS: Our data suggest that the phosphorylation of Thr144 in CaMKKβ is rapidly induced by cAMP/cAMP-dependent protein kinase (PKA) signaling in CaMKKβ-transfected HeLa cells, that is physiologically relevant in mouse cerebellum. We confirmed that the catalytic subunit of PKA was capable of directly phosphorylating CaMKKβ at Thr144 in vitro and in transfected cells. In addition, the basal phosphorylation of CaMKKβ at Thr144 in transfected HeLa cells was suppressed by AMPK inhibitor (compound C). PKA-catalyzed phosphorylation reduced the autonomous activity of CaMKKβ in vitro without significant effect on the Ca2+/CaM-dependent activity, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme. CONCLUSION: cAMP/PKA signaling may confer Ca2+-dependency to the CaMKKβ-mediated signaling pathway through direct phosphorylation of Thr144 in intact cells. GENERAL SIGNIFICANCE: Our results suggest a novel cross-talk between cAMP/PKA and Ca2+/CaM/CaMKKβ signaling through regulatory phosphorylation. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 00201693 484 2018 Syntheses and crystal structures of neodymium(III) and europium(III) complexes bearing dimethyl-, pyrrolidine-, or S-prolinol- dithiocarbamato ligands and their natural and magnetic circular dichroism spectra 394 401 EN Abdallah Yakubu Graduate School of Natural Science and Technology, Okayama University Takayoshi Suzuki Graduate School of Natural Science and Technology, Okayama University Masakazu Kita Faculty of Education, Okayama University A series of Nd-III and Eu-III complexes containing achiral or chiral dithiocarbamato (dtc) ligands, [Ln(Xdtc)(3)(NN)] {Ln = Nd or Eu; X = dimethyl- (Me-2), pyrrolidine- (pyr), or (S)-prolinol- (S-proOH); NN = 1,10-phenanthroline (phen) or 2,2'-bipyridine (bpy)}, were prepared and their crystal structures and spectroscopic properties, in particular the natural circular dichroism (CD) and magnetic circular dichroism (MCD), were investigated. The crystal structures of the complexes analyzed by the X-ray diffraction method showed an 8-coordinate geometry around the Ln III center with comparable structural parameters to one another and to the related complexes reported previously. These complexes exhibited similar spectral patterns in their absorption, natural CD and MCD spectra in solution. Weak but characteristic sharp f-f transition bands were observed in the absorption and MCD spectra, but no CD signals associated with these transitions were observed even in the S-proOHdtc complexes. The MCD spectral pattern of the Eu-III complexes revealed a local C-2v symmetry around the Ln(III) center in solution, in contrast to the aqua and the analogous beta-diketonato Eu-III complexes. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 23523964 25 2017 Attenuation of CD4+ CD25+ Regulatory T Cells in the Tumor Microenvironment by Metformin, a Type 2 Diabetes Drug 154 164 EN Yuki Kunisada Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shingo Eikawa Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Nahoko Tomonobu Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shohei Domae Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takenori Uehara Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shohei Hori Laboratory of Immunology and Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo Yukihiro Furusawa Division of Biochemistry, Keio University Graduate School of Pharmaceutical Science Koji Hase Division of Biochemistry, Keio University Graduate School of Pharmaceutical Science Akira Sasaki Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Heiichiro Udono Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences CD4+CD25+ regulatory T cells (Treg), an essential subset for preventing autoimmune diseases, is implicated as a negative regulator in anti-tumor immunity. We found that metformin (Met) reduced tumor-infiltrating Treg (Ti-Treg), particularly the terminally-differentiated CD103+KLRG1+ population, and also decreased effector molecules such as CTLA4 and IL-10. Met inhibits the differentiation of naïve CD4+ T cells into inducible Treg (iTreg) by reducing forkhead box P3 (Foxp3) protein, caused by mTORC1 activation that was determined by the elevation of phosphorylated S6 (pS6), a downstream molecule of mTORC1. Rapamycin and compound C, an inhibitor of AMP-activated protein kinase (AMPK) restored the iTreg generation, further indicating the involvement of mTORC1 and AMPK. The metabolic profile of iTreg, increased Glut1-expression, and reduced mitochondrial membrane-potential and ROS production of Ti-Treg aided in identifying enhanced glycolysis upon Met-treatment. The negative impact of Met on Ti-Treg may help generation of the sustained antitumor immunity. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 09201211 105 1-2 2013 Inhalation of 10% carbon dioxide rapidly terminates Scn1a mutation-related hyperthermia-induced seizures 220 224 EN Iori Ohmori Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Keiichiro Hayashi Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Haijiao Wang Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Mamoru Ouchida Department of Molecular Genetics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Naohiro Fujita Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takushi Inoue Department of Child Neurology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroyuki Michiue Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Teiichi Nishiki Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hideki Matsui Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University The aim of this study was to assess the anticonvulsant effect of carbon dioxide (CO2) on Scn1a mutation-related febrile seizures. We examined physiological changes in the blood gas levels after the induction of hyperthermia-induced seizures (HISs), which were associated with the Scn1a missense mutation. We determined the efficacy of inhalation of 5% or 10% CO2 to treat HISs. HISs were evoked in Scn1a mutant and wild-type (WT) rats by hot water baths. To determine the anticonvulsant effect of CO2 inhalation, rats were placed in a chamber filled with air or mixed gas containing 5% CO2 or 10% CO2 for 3 min, immediately after the induction of HISs. We also analyzed the blood gas levels at the end of inhalation of CO2. Hot water bathing induced a significant reduction in the partial pressure of CO2 (pCO2) and respiratory alkalosis in the WT and Scn1a mutant rats. HISs were evoked in 100% of the Scn1a mutant rats within 5 min, but in none of the WT rats. The Scn1a mutant rats demonstrated a higher HISs susceptibility associated with respiratory alkalosis than the WT rats. Inhalation of 10% CO2 shortened the seizure duration from 62.6±12.1 s to 15.5±1.0 s. Blood gas analysis after the inhalation of 10% CO2 demonstrated an elevated pCO2 level and respiratory acidosis. Inhalation of 10% CO2 demonstrated a potent and fast-acting anticonvulsant effect against HISs. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 01689002 913 2019 Energy response of X-rays under high flux conditions using a thin APD for the energy range of 6–33 keV 72 77 EN T. Masuda Research Institute for Interdisciplinary Science, Okayama University T. Hiraki Research Institute for Interdisciplinary Science, Okayama University H. Kaino Research Institute for Interdisciplinary Science, Okayama University S. Kishimoto High Energy Accelerator Research Organization Y. Miyamoto Research Institute for Interdisciplinary Science, Okayama University K. Okai Research Institute for Interdisciplinary Science, Okayama University S. Okubo Research Institute for Interdisciplinary Science, Okayama University R. Ozaki Research Institute for Interdisciplinary Science, Okayama University N. Sasao Research Institute for Interdisciplinary Science, Okayama University K. Suzuki Research Institute for Interdisciplinary Science, Okayama University S. Uetake Research Institute for Interdisciplinary Science, Okayama University A. Yoshimi Research Institute for Interdisciplinary Science, Okayama University K. Yoshimura Research Institute for Interdisciplinary Science, Okayama University This paper reports on the demonstration of a high-rate energy measurement technique using a thin depletion layer silicon avalanche photodiode (Si-APD). A dedicated amplitude-to-time converter is developed to realize simultaneous energy and timing measurement in a high rate condition. The energy response of the system is systematically studied by using monochromatic X-ray beam with an incident energy ranging from 6 to 33 keV. The obtained energy spectra contain clear peaks and tail distributions. The peak fraction monotonously decreases as the incident photon energy increases. This phenomenon can be explained by considering the distribution of the energy deposit in silicon, which is investigated by using a Monte Carlo simulation. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 01677322 200 Part.A 2014 A model-free method for extracting interaction potential between protein molecules using small-angle X-ray 42 46 EN Tomonari Sumi Department of Chemistry, Faculty of Science, Okayama University Hiroshi Imamura Graduate School of Advanced integration Science, Chiba University Takeshi Morita Graduate School of Advanced integration Science, Chiba University Keiko Nishikawa Graduate School of Advanced integration Science, Chiba University A small-angle X-ray scattering has been used to probe protein–protein interaction in solution. Conventional methods need to input modeled potentials with variable/invariable parameters to reproduce the experimental structure factor. In the present study, a model-free method for extracting the excess part of effective interaction potential between protein molecules in solutions over an introduced hard-sphere potential by using experimental data of small-angle X-ray scattering is presented on the basis of liquid-state integral equation theory. The reliability of the model-free method is tested by the application to experimentally derived structure factors for dense lysozyme solutions with different solution conditions [Javid et al., Phys. Rev. Lett. 99, 028101 (2007), Schroer et al., Phys. Rev. Lett. 106, 178102 (2011)]. The structure factors calculated from the model-free method agree well with the experimental ones. The model-free method provides the following picture of the lysozyme solution: these are the stabilization of contact-pair configurations, large activation barrier against their formations, and screened Coulomb repulsion between the charged proteins. In addition, the model-free method will be useful to verify whether or not a model for colloidal system is acceptable to describing protein–protein interaction. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 01681702 177 1 2013 Characterization of burdock mottle virus, a novel member of the genus Benyvirus, and the identification of benyvirus-related sequences in the plant and insect genomes. 75 86 EN Hideki Kondo Institute of Plant Science and Resources (IPSR), Okayama University Shuichi Hirano Institute of Plant Science and Resources (IPSR), Okayama University Sotaro Chiba Institute of Plant Science and Resources (IPSR), Okayama University Ida Bagus Andika Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences Makoto Hirai Department of Parasitology, Graduate School of Medicine, Gunma University Takanori Maeda Formerly College of Bioresource Sciences, Nihon University Tetsuo Tamada Institute of Plant Science and Resources (IPSR), Okayama University The complete nucleotide sequence of the burdock mottle virus (BdMoV) isolated from an edible burdock plant (Arctium lappa) in Japan has been determined. BdMoV has a bipartite genome, whose organization is similar to RNA1 and RNA2 of benyviruses, beet necrotic yellow vein virus (BNYVV), beet soil-borne mosaic virus (BSBMV), and rice stripe necrosis virus (RSNV). BdMoV RNA1 (7038 nt) contains a single open reading frame (ORF) encoding a 249-kDa polypeptide that consists of methyl-transferase, helicase, papain-like protease, AlkB-like, and RNA-dependent RNA polymerase domains. The AlkB-like domain sequence is not present in the proteins encoded by other known benyviruses, but is found in replication-associated proteins of viruses mainly belonging to the families Alfaflexiviridae and Betaflexiviridae. BdMoV RNA2 (4315 nt) contains six ORFs that are similar to those of benyviruses: these are coat protein (CP), CP readthrough, triple gene block movement and cysteine-rich proteins. Phylogenetic analyses showed that BdMoV is more closely related to BNYVV and BSBMV than to RSNV. Database searches showed that benyvirus replicase-related sequences are present in the chromosomes of a chickpea plant (Cicer arietinum) and a blood-sucking insect (Rhodnius prolixus). Some other benyvirus-related sequences are found in the transcriptome shotgun libraries of a few species of plants and a bark beetle. Our results show that BdMoV is a distinct species of the genus Benyvirus and that ancestral and extant benyviruses may have infected or currently infect a wide range of hosts, including plants and insects. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 01681702 213 2016 Sequence and phylogenetic analyses of novel totivirus-like double-stranded RNAs from field-collected powdery mildew fungi 353 364 EN Hideki Kondo Institute of Plant Science and Resources (IPSR), Okayama University Sakae Hisano Institute of Plant Science and Resources (IPSR), Okayama University Sotaro Chiba Institute of Plant Science and Resources (IPSR), Okayama University Kazuyuki Maruyama Institute of Plant Science and Resources (IPSR), Okayama University Ida Bagus Andika Institute of Plant Science and Resources (IPSR), Okayama University Kazuhiro Toyoda Graduate School of Environmental and Life Science, Okayama University Fumihiro Fujimori Department of Environmental Education, Tokyo Kasei University Nobuhiro Suzuki Institute of Plant Science and Resources (IPSR), Okayama University 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. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 24058440 5 8 2019 Effectiveness and safety of low-concentrated ozonized water for the reduction of contamination in dental unit water lines e02306 EN Keisuke Okubo Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takashi Ito Center for Innovative Clinical Medicine, Okayama University Hospital Yasuyoshi Shiota Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yusuke Kawata Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tadashi Yamamoto Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shogo Takashiba Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Contamination of dental unit waterlines (DUWL) with heterotrophic bacteria can cause problems in immune compromised patients (aged, tumor and organ transplantation-patients). We focused on the use of low-concentrated ozonized water (OZW) as the biofilm formation restraint system for DUWL. Here, we examined the effects of low-concentrated OZW on the growth of bacteria and related biofilm formation and harmfulness to dental unit components (DUCs) in vitro. Objectives To evaluate the bactericidal effects of OZW on biofilms in DUWL and DUC in vitro. Methods Low-concentrated OZW (0.4 mg/L) was generated using an OZS-PTDX generator. Heterotrophic bacterial biofilms in old DUWL tubes and Candia albicans solution (control microbe) were treated with OZW for 1 h with gentle agitation before static culturing for 96 h in Reasoner's 2A liquid media. The control solutions were 0.1% cetylpyridinium chloride (CPC), chlorinated tap water (TW), and phosphate-buffered saline (PBS). Adenosine triphosphate (ATP) amounts of the microbes were measured and the biofilms of these microbes were observed using scanning electron microscopy (SEM). Moreover, surfaces of DUC soaked in OZW and TW were observed by SEM. Results The OZW reduced ATP levels in microbes to 50% compared to TW and PBS treatment, although CPC reduced it below detection limits. SEM observation revealed deformation of microbes cultured with OZW, whereas no changes were seen on DUC surfaces. Conclusions Low-concentrated OZW is bactericidal against heterotrophic bacteria biofilms and it is not harmful to DUC, suggesting that it might be useful in preventing DUWL contamination. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 24058440 5 6 2019 Vision evaluation by functional observational battery, operant behavior test, and light/dark box test in retinal dystrophic RCS rats versus normal rats e01936 EN Toshihiko Matsuo Ophthalmology, Okayama University Medical School and Okayama University Hospital Tetsuya Uchida Polymer Materials Science, Okayama University Graduate School of Natural Science and Technology Koichiro Yamashita Polymer Materials Science, Okayama University Graduate School of Natural Science and Technology Shigiko Takei Ina Research Daisuke Ido Ina Research Atsushi Fujiwara Ina Research Masahiko Iino Ina Research Masao Oguchi Ina Research BACKGROUND: Vision plays a key role in some behavior tests for rats. Okayama University-type retinal prosthesis (OUReP) is a photoelectric dye-coupled polyethylene film which generates electric potential in response to light and stimulates nearby neurons. This study aims to assess vision in retinal dystrophic (RCS) rats, in comparison with normal rats, by selected behavior tests. We also examined whether the tests could detect vision changes in RCS rats with dye-coupled film implantation. METHODS: Data sets were 5 normal rats, 4 untreated RCS rats, 7 RCS rats with dye-coupled films implanted at the age of 7 weeks after excluding unsuccessful implantation at autopsy. Behavior tests chosen were landing foot splay and visual forelimb-placing response in the menu of functional observational battery, operant-conditioning lever-press response and light/dark box test. RESULTS: Normal visual placing response was significantly less frequent in untreated RCS rats at the age of 9 and 11 weeks, compared with normal rats (P = 0.0027, chi-square test) while normal response was significantly more frequent at the age of 9 weeks in RCS rats with dye-coupled film implantation, compared with untreated RCS rats (P = 0.0221). In operant-conditioning lever-press test, the correct response rate was significantly lower in untreated RCS rats than in normal rats at the age of 9 weeks (P < 0.05, Tukey-Kramer test) while the rate was not significantly different between normal rats and RCS rats with dye-coupled film implantation. In light/dark box test, the time to enter dark box was significantly shorter in normal rats, compared with untreated RCS rats or RCS rats with dye-coupled film implantation (P < 0.05, Tukey-Kramer test). CONCLUSIONS: Behavior tests of functional observational battery, operant-conditioning lever-press response and light/dark box test discriminated vision between normal rats and RCS rats. The visual placing response and operant-conditioning lever-press test might have sensitivity to detect vision recovery in RCS rats with OUReP implantation. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 15423565 17 6 2019 Myoepithelial Hamartoma in the Ampulla of Vater e56 e57 EN Takeshi Tomoda Department of Gastroenterology, Okayama University Hospital Hironari Kato Department of Gastroenterology, Okayama University Hospital Hiroyuki Okada Department of Gastroenterology, Okayama University Hospital No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 01681702 262 2019 A novel insect-infecting virga/nege-like virus group and its pervasive endogenization into insect genomes 37 47 EN Hideki Kondo Institute of Plant Science and Resources (IPSR), Okayama University Sotaro Chiba Asian Satellite Campuses Institute, Nagoya University Kazuyuki Maruyama Institute of Plant Science and Resources (IPSR), Okayama University Ida Bagus Andika Institute of Plant Science and Resources (IPSR), Okayama University Nobuhiro Suzuki Institute of Plant Science and Resources (IPSR), Okayama University Insects are the host and vector of diverse viruses including those that infect vertebrates, plants, and fungi. Recent wide-scale transcriptomic analyses have uncovered the existence of a number of novel insect viruses belonging to an alphavirus-like superfamily (virgavirus/negevirus-related lineage). In this study, through an in silico search using publicly available insect transcriptomic data, we found numerous virus-like sequences related to insect virga/nege-like viruses. Phylogenetic analysis showed that these novel viruses and related virus-like sequences fill the major phylogenetic gaps between insect and plant virga/negevirus lineages. Interestingly, one of the phylogenetic clades represents a unique insect-infecting virus group. Its members encode putative coat proteins which contained a conserved domain similar to that usually found in the coat protein of plant viruses in the family Virgaviridae. Furthermore, we discovered endogenous viral elements (EVEs) related to virga/nege-like viruses in the insect genomes, which enhances our understanding on their evolution. Database searches using the sequence of one member from this group revealed the presence of EVEs in a wide range of insect species, suggesting that there has been prevalent infection by this virus group since ancient times. Besides, we present detailed EVE integration profiles of this virus group in some species of the Bombus genus of bee families. A large variation in EVE patterns among Bombus species suggested that while some integration events occurred after the species divergence, others occurred before it. Our analyses support the view that insect and plant virga/nege-related viruses might share common virus origin(s). No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 15671348 54 2017 Characterization of Vibrio cholerae O1 strains that trace the origin of Haitian-like genetic traits 47 53 EN Priyanka Ghosh Division of Bacteriology, National Institute of Cholera and Enteric Diseases Dhirendra Kumar Maharishi Valmiki Infectious Diseases Hospital Goutam Chowdhury Division of Bacteriology, National Institute of Cholera and Enteric Diseases Puneeta Singh Maharishi Valmiki Infectious Diseases Hospital Prosenjit Samanta Division of Bacteriology, National Institute of Cholera and Enteric Diseases Shanta Dutta Division of Bacteriology, National Institute of Cholera and Enteric Diseases T. Ramamurthy Division of Bacteriology, National Institute of Cholera and Enteric Diseases N. C. Sharma Maharishi Valmiki Infectious Diseases Hospital Preety Sinha Department of Zoology, A.N. College Yogendra Prasad Department of Animal Science, MJP Rohilkhand University Sumio Shinoda Collaborative Research Center of Okayama University for Infectious Diseases at NICED Asish K. Mukhopadhyay Division of Bacteriology, National Institute of Cholera and Enteric Diseases Vibrio cholerae O1 is the etiological agent of the severe diarrheal disease cholera. The bacterium has recently been causing outbreaks in Haiti with catastrophic effects. Numerous mutations have been reported in V. cholerae O1 strains associated with the Haitian outbreak. These mutations encompass among other the genes encoding virulence factors such as the pilin subunit of the toxin-co-regulated pilus (tcpA), cholera toxin B subunit (ctxB), repeat in toxins (rtxA), and other genes such as the quinolone resistance-determining region (QRDR) of gyrase A (gyrA), rstB of RS element along with the alteration in the number of repeat sequences at the promoter region of ctxAB. Given the numerous genetic changes in those Haitian isolates, we decided to investigate the possible origins of those variations in the Indian subcontinent. Thus, we determined the genetic traits among V. cholerae O1 strains in Delhi, India. A total of 175 strains isolated from cholera patients during 2004 to 2012 were analysed in the present study. Our results showed that all the tested strains carried Haitian type tcpA (tcpACIRS) and variant gyrA indicating their first appearance before 2004 in Delhi. The Haitian variant rtxA and ctxB7 were first detected in Delhi during 2004 and 2006, respectively. Interestingly, not a single strain with the combination of El Tor rtxA and ctxB7 was detected in this study. The Delhi strains carried four heptad repeats (TTTTGAT) in the CT promoter region whereas Haitian strains carried 5 such repeats. Delhi strains did not have any deletion mutations in the rstB like Haitian strains. Overall, our study demonstrates the sequential accumulation of Haitian-like genetic traits among V. cholerae O1 strains in Delhi at different time points prior to the Haitian cholera outbreak. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 0264410X 32 supplment 1 2014 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 A20 A28 EN Satarupa Mullick National Institute of Cholera and Enteric Diseases Paulami Mandal National Institute of Cholera and Enteric Diseases Mukti Kant Nayak National Institute of Cholera and Enteric Diseases Souvik Ghosh Department of Hygiene, Sapporo Medical University School of Medicine Papiya De National Institute of Cholera and Enteric Diseases K. Rajendran National Institute of Cholera and Enteric Diseases Mihir K. Bhattacharya National Institute of Cholera and Enteric Diseases Utpala Mitra National Institute of Cholera and Enteric Diseases Thandavarayan Ramamurthy National Institute of Cholera and Enteric Diseases Nobumichi Kobayashi Department of Hygiene, Sapporo Medical University School of Medicine Mamta Chawla-Sarkar National Institute of Cholera and Enteric Diseases 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. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 0925-5710 105 6 2017 Predictors of vasovagal reactions during preoperative autologous blood donation: a single-institution analysis 812 818 EN Hisakazu Nishimori Department of Transfusion Medicine, Okayama University Hospital Nobuharu Fujii Department of Transfusion Medicine, Okayama University Hospital Keiko Fujii Department of Transfusion Medicine, Okayama University Hospital Tohru Ikeda Department of Transfusion Medicine, Okayama University Hospital Naomi Asano Department of Transfusion Medicine, Okayama University Hospital Hiroaki Ogo Department of Transfusion Medicine, Okayama University Hospital Miwa Yamakawa Department of Nursing, Okayama University Hospital Naoe Takagi Department of Nursing, Okayama University Hospital Fumio Otsuka Department of Transfusion Medicine, Okayama University Hospital Kazuma Ikeda Department of Transfusion Medicine, Okayama University Hospital Studies examining risk factors associated with vasovagal reactions (VVRs) during autologous blood donations, especially in younger subjects, have been limited. The aim of the present study was to define risk factors for VVRs during preoperative autologous blood donation in patients, including those younger than 18 years old. We retrospectively analyzed 4192 autologous, preoperative blood donations between 2007 and 2015 at Okayama University Hospital. Eighty-seven (2.08%) of the patients experienced VVRs. VVRs occurred approximately three times as often in patients 0-17 years old (16/320, 5.0%) than in patients 18 years and older (71/3872, 1.8%). In particular, VVRs occurred more frequently in those 10-13 years old, and decreased with older age (P = 0.006). In a univariate analysis, younger age, lower body mass index, lower systolic blood pressure, lower body weight, lower total blood volume, female gender, first-time collection, and higher heart rate were associated with a higher incidence of VVRs. In a multivariate analysis, lower systolic blood pressure (P < 0.001), higher heart rate (P = 0.007), and first-time collection (P = 0.015), remained independent predictors of VVRs. These results emphasize the need for careful attention during blood collection. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 0109-5641 33 3 2017 Sandblasting may damage the surface of composite CAD-CAM blocks e124 e135 EN Kumiko Yoshihara Centrer for Innovative Clinical Medicine, Okayama University Hospital Noriyuki Nagaoka Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School Yukinori Maruo Department of Occlusion and Removable Prosthodontics, Okayama University Hospital Goro Nishigawa Department of Occlusion and Removable Prosthodontics, Okayama University Hospital Masao Irie Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuhiro Yoshida Department of Biomaterials and Bioengineering, Graduate School of Dental Medicine, Hokkaido University Bart Van Meerbeekf KU Leuven (University of Leuven), Department of Oral Health Sciences, BIOMAT & University Hospitals 　OBJECTIVE:　 CAD-CAM blocks to fabricate semi-direct and indirect restorations are available in different sorts of ceramics as well as composite. In order to bond restorations prepared out of composite blocks into tooth cavities, it is recommended to gently sandblast the surface prior to the application of a primer/adhesive. Today, the effect of sandblasting composite block surfaces has not thoroughly been investigated. In this study, the ultra-structure of composite CAD-CAM blocks was investigated with special attention to the effect of sandblasting on the surface topography and of silanization on the bonding performance. 　METHODS:　 Five different composite CAD-CAM blocks were involved. We correlatively investigated their structural and chemical composition using X-ray diffraction (XRD), energy dispersion spectroscopy (EDS), scanning electron microscopy (SEM) and (scanning) transmission electron microscopy ((S)TEM). The effect of sandblasting was also imaged in cross-section and at the interface with composite cement. Finally, we measured the shear bond strength to the sandblasted block surface with and without silanization. 　RESULTS:　 All composite blocks revealed a different ultra-structure. Sandblasting increased surface roughness and resulted in an irregular surface with some filler exposure. Sandblasting also damaged the surface. When the sandblasted composite blocks were silanized, superior bonding receptiveness in terms of higher bond strength was achieved except for Shofu Block HC. 　SIGNIFICANCE:　 Sandblasting followed by silanization improved the bond strength to composite CAD-CAM blocks. However, sandblasting may also damage the composite CAD-CAM block surface. For the composite CAD-CAM block Shofu Block HC, the damage was so severe that silanization did not improve bond strength. No potential conflict of interest relevant to this article was reported.

Elsevier Science Acta Medica Okayama 0168-1702 167 1 2012 Identification of host genes showing differential expression profiles with cell-based long-term replication of hepatitis C virus RNA 74 85 EN Hiroe Sejima Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Kyoko Mori Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yasuo Ariumi Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Masanori Ikeda Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Nobuyuki Kato Department of Tumor Virology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences 　Persistent hepatitis C virus (HCV) infection frequently causes hepatocellular carcinoma. However, the mechanisms of HCV-associated hepatocarcinogenesis and disease progression are unclear. Although the human hepatoma cell line, HuH-7, has been widely used as the only cell culture system for robust HCV replication, we recently developed new human hepatoma Li23 cell line-derived OL, OL8, OL11, and OL14 cells, in which genome-length HCV RNA (O strain of genotype 1b) efficiently replicates. OL, OL8, OL11, and OL14 cells were cultured for more than 2 years. We prepared cured cells from OL8 and OL11 cells by interferon-γ treatment. The cured cells were also cultured for more than 2 years. cDNA microarray and RT-PCR analyses were performed using total RNAs prepared from these cells. We first selected several hundred highly or moderately expressed probes, the expression levels of which were upregulated or downregulated at ratios of more than 2 or less than 0.5 in each set of compared cells (e.g., parent OL8 cells versus OL8 cells cultured for 2 years). From among these probes, we next selected those whose expression levels commonly changed during a 2-year culture of genome-length HCV RNA-replicating cells, but which did not change during a 2-year culture period in cured cells. We further examined the expression levels of the selected candidate genes by RT-PCR analysis using additional specimens from the cells cultured for 3.5 years. Reproducibility of the RT-PCR analysis using specimens from recultured cells was also confirmed. Finally, we identified 5 upregulated genes and 4 downregulated genes, the expression levels of which were irreversibly altered during 3.5-year replication of HCV RNA. These genes may play roles in the optimization of the environment in HCV RNA replication, or may play key roles in the progression of HCV-associated hepatic diseases. No potential conflict of interest relevant to this article was reported.