Frontiers MediaActa Medica Okayama1662-5196162023A method for reconstruction of interpretable brain networks from transient synchronization in resting-state BOLD fluctuations960607ENYusukeNoroDepartment of Biosciences and Informatics, Keio UniversityRuixiangLiDepartment of Physiology, The University of Tokyo School of MedicineTeppeiMatsuiDepartment of Biology, Okayama UniversityKojiJimuraDepartment of Informatics, Gunma UniversityResting-state (rs) fMRI has been widely used to examine brain-wide large-scale spatiotemporal architectures, known as resting-state networks (RSNs). Recent studies have focused on the temporally evolving characteristics of RSNs, but it is unclear what temporal characteristics are reflected in the networks. To address this issue, we devised a novel method for voxel-based visualization of spatiotemporal characteristics of rs-fMRI with a time scale of tens of seconds. We first extracted clusters of dominant activity-patterns using a region-of-interest approach and then used these temporal patterns of the clusters to obtain voxel-based activation patterns related to the clusters. We found that activation patterns related to the clusters temporally evolved with a characteristic temporal structure and showed mutual temporal alternations over minutes. The voxel-based representation allowed the decoding of activation patterns of the clusters in rs-fMRI using a meta-analysis of functional activations. The activation patterns of the clusters were correlated with behavioral measures. Taken together, our analysis highlights a novel approach to examine brain activity dynamics during rest.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-302X122021Comprehensive Comparative Genomics and Phenotyping of Methylobacterium Species740610ENOlaAlessaInstitute of Plant Science and Resources, Okayama UniversityYoshitoshiOguraDivision of Microbiology, Department of Infectious Medicine, Kurume University School of MedicineYoshikoFujitaniInstitute of Plant Science and Resources, Okayama UniversityHidetoTakamiAtmosphere and Ocean Research Institute, The University of TokyoTetsuyaHayashiDepartment of Bacteriology, Graduate School of Medical Sciences, Kyushu UniversityNurettinSahinEgitim Fakultesi, Mugla Sitki Kocman UniversityAkioTaniInstitute of Plant Science and Resources, Okayama UniversityThe pink-pigmented facultative methylotrophs (PPFMs), a major bacterial group found in the plant phyllosphere, comprise two genera: Methylobacterium and Methylorubrum. They have been separated into three major clades: A, B (Methylorubrum), and C. Within these genera, however, some species lack either pigmentation or methylotrophy, which raises the question of what actually defines the PPFMs. The present study employed a comprehensive comparative genomics approach to reveal the phylogenetic relationship among the PPFMs and to explain the genotypic differences that confer their different phenotypes. We newly sequenced the genomes of 29 relevant-type strains to complete a dataset for almost all validly published species in the genera. Through comparative analysis, we revealed that methylotrophy, nitrate utilization, and anoxygenic photosynthesis are hallmarks differentiating the PPFMs from the other Methylobacteriaceae. The Methylobacterium species in clade A, including the type species Methylobacterium organophilum, were phylogenetically classified into six subclades, each possessing relatively high genomic homology and shared phenotypic characteristics. One of these subclades is phylogenetically close to Methylorubrum species; this finding led us to reunite the two genera into a single genus Methylobacterium. Clade C, meanwhile, is composed of phylogenetically distinct species that share relatively higher percent G+C content and larger genome sizes, including larger numbers of secondary metabolite clusters. Most species of clade C and some of clade A have the glutathione-dependent pathway for formaldehyde oxidation in addition to the H4MPT pathway. Some species cannot utilize methanol due to their lack of MxaF-type methanol dehydrogenase (MDH), but most harbor an XoxF-type MDH that enables growth on methanol in the presence of lanthanum. The genomes of PPFMs encode between two and seven (average 3.7) genes for pyrroloquinoline quinone-dependent alcohol dehydrogenases, and their phylogeny is distinctly correlated with their genomic phylogeny. All PPFMs were capable of synthesizing auxin and did not induce any immune response in rice cells. Other phenotypes including sugar utilization, antibiotic resistance, and antifungal activity correlated with their phylogenetic relationship. This study provides the first inclusive genotypic insight into the phylogeny and phenotypes of PPFMs.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1662-5196152022Counterfactual Explanation of Brain Activity Classifiers Using Image-To-Image Transfer by Generative Adversarial Network802938ENTeppeiMatsuiDepartment of Biology, Okayama UniversityMasatoTaki Graduate School of Artificial Intelligence and Science, Rikkyo UniversityTrung QuangPhamSupportive Center for Brain Research, National Institute for Physiological SciencesJunichiChikazoeSupportive Center for Brain Research, National Institute for Physiological SciencesKojiJimuraDepartment of Biosciences and Informatics, Keio UniversityDeep neural networks (DNNs) can accurately decode task-related information from brain activations. However, because of the non-linearity of DNNs, it is generally difficult to explain how and why they assign certain behavioral tasks to given brain activations, either correctly or incorrectly. One of the promising approaches for explaining such a black-box system is counterfactual explanation. In this framework, the behavior of a black-box system is explained by comparing real data and realistic synthetic data that are specifically generated such that the black-box system outputs an unreal outcome. The explanation of the system's decision can be explained by directly comparing the real and synthetic data. Recently, by taking advantage of advances in DNN-based image-to-image translation, several studies successfully applied counterfactual explanation to image domains. In principle, the same approach could be used in functional magnetic resonance imaging (fMRI) data. Because fMRI datasets often contain multiple classes (e.g., multiple behavioral tasks), the image-to-image transformation applicable to counterfactual explanation needs to learn mapping among multiple classes simultaneously. Recently, a new generative neural network (StarGAN) that enables image-to-image transformation among multiple classes has been developed. By adapting StarGAN with some modifications, here, we introduce a novel generative DNN (counterfactual activation generator, CAG) that can provide counterfactual explanations for DNN-based classifiers of brain activations. Importantly, CAG can simultaneously handle image transformation among all the seven classes in a publicly available fMRI dataset. Thus, CAG could provide a counterfactual explanation of DNN-based multiclass classifiers of brain activations. Furthermore, iterative applications of CAG were able to enhance and extract subtle spatial brain activity patterns that affected the classifier's decisions. Together, these results demonstrate that the counterfactual explanation based on image-to-image transformation would be a promising approach to understand and extend the current application of DNNs in fMRI analyses.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-462X112021Cytosolic Free N-Glycans Are Retro-Transported Into the Endoplasmic Reticulum in Plant Cells610124ENMakotoKatsubeDepartment of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama UniversityNatsukiEbaraDepartment of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama UniversityMegumiMaedaDepartment of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama UniversityYoshinobuKimuraDepartment of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama UniversityDuring endoplasmic reticulum (ER)-associated degradation, free N-glycans (FNGs) are produced from misfolded nascent glycoproteins via the combination of the cytosolic peptide N-glycanase (cPNGase) and endo-beta-N-acetylglucosaminidase (ENGase) in the plant cytosol. The resulting high-mannose type (HMT)-FNGs, which carry one GlcNAc residue at the reducing end (GN1-FNGs), are ubiquitously found in developing plant cells. In a previous study, we found that HMT-FNGs assisted in protein folding and inhibited beta-amyloid fibril formation, suggesting a possible biofunction of FNGs involved in the protein folding system. However, whether these HMT-FNGs occur in the ER, an organelle involved in protein folding, remained unclear. On the contrary, we also reported the presence of plant complex type (PCT)-GN1-FNGs, which carry the Lewis(a) epitope at the non-reducing end, indicating that these FNGs had been fully processed in the Golgi apparatus. Since plant ENGase was active toward HMT-N-glycans but not PCT-N-glycans that carry beta 1-2xylosyl and/or alpha 1-3 fucosyl residue(s), these PCT-GN1-FNGs did not appear to be produced from fully processed glycoproteins that harbored PCT-N-glycans via ENGase activity. Interestingly, PCT-GN1-FNGs were found in the extracellular space, suggesting that HMT-GN1-FNGs formed in the cytosol might be transported back to the ER and processed in the Golgi apparatus through the protein secretion pathway. As the first step in elucidating the production mechanism of PCT-GN1-FNGs, we analyzed the structures of free oligosaccharides in plant microsomes and proved that HMT-FNGs (Man(9-7)GlcNAc(1) and Man(9-8)GlcNAc(2)) could be found in microsomes, which almost consist of the ER compartments.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224112021Deletion of Mir223 Exacerbates Lupus Nephritis by Targeting S1pr1 in Fas(lpr/lpr) Mice616141ENSumieHiramatsu-AsanoDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityKatsueSunahori-WatanabeDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversitySoniaZeggarDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityEriKatsuyamaDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTomoyukiMukaiDepartment of Rheumatology, Kawasaki Medical SchoolYoshitakaMoritaDepartment of Rheumatology, Kawasaki Medical SchoolJunWadaDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityObjective: The micro RNAs (miRNAs) and their target mRNAs are differentially expressed in various immune-mediated cells. Here, we investigated the role of Mir223 and sphingosine-1-phosphate receptor 1 (S1pr1) in the pathogenesis of systemic lupus erythematosus.</br>
Methods: We analyzed miRNA and mRNA profiling data of CD4+ splenic T cells derived from MRL/MpJ-Faslpr/J mice. We performed 3′ untranslated region (UTR) luciferase reporter gene assay using human umbilical vein endothelial cells (HUVECs). We generated the B6-Mir223−/−Faslpr/lpr mice and the lupus phenotypes were analyzed.</br>
Results: In CD4+ splenic T cells, we identified upregulation of miR-223-3p and downregulation of the possible target, S1pr1 by RNA sequencing of MRL/MpJ-Faslpr/J mice. The transfection with miR-223-3p mimic significantly suppressed a luciferase activity in HUVEC treated with a Lentivirus vector containing 3′ UTR of S1pr1. The mRNA levels of S1pr1 were significantly decreased after miR-223-3p overexpression. In B6-Mir223−/−Faslpr/lpr mice, the proportion of CD3+ T cells, CD3+CD4-CD8− cells, B cells, plasma cells, and S1PR1+CD4+ T cells in the spleen was significantly increased compared with that in B6-Mir223+/+Faslpr/lpr mice by flow cytometry. B6-Mir223−/−Faslpr/lpr mice demonstrated the elevation of glomerular and renal vascular scores associated with enhanced intraglomerular infiltration of S1PR1+CD4+ T cells.</br>
Conclusion: Unexpectedly, the deletion of Mir223 exacerbated the lupus phenotypes associated with increased population of S1PR1+CD4+ T in spleen and the enhanced infiltration of S1PR1+CD4+ T cells in inflamed kidney tissues, suggesting compensatory role of Mir223 in the pathogenesis of lupus nephritis.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1662-5196142020Development of a Non-invasive Deep Brain Stimulator With Precise Positioning and Real-Time Monitoring of Bioimpedance574189ENHengWangSchool of Mechatronic Engineering, Beijing Institute of TechnologyZhongyanShiSchool of Life Science, Beijing Institute of TechnologyWeiqianSunSchool of Life Science, Beijing Institute of TechnologyJianxuZhangSchool of Mechatronic Engineering, Beijing Institute of TechnologyJingWangDepartment of Health Management, Aerospace Center Hospital, Peking University Aerospace School of Clinical MedicineYueShiBeijing Big-IQ Medical Equipment Co., Ltd.RuoshuiYangSchool of Mechatronic Engineering, Beijing Institute of TechnologyChunlinLiSchool of Biomedical Engineering, Capital Medical UniversityDuanduanChenSchool of Life Science, Beijing Institute of TechnologyJinglongWuSchool of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityGuoGongyaoSchool of Life Science, Beijing Institute of TechnologyYifeiXuSchool of Life Science, Beijing Institute of TechnologyMethods by which to achieve non-invasive deep brain stimulation via temporally interfering with electric fields have been proposed, but the precision of the positioning of the stimulation and the reliability and stability of the outputs require improvement. In this study, a temporally interfering electrical stimulator was developed based on a neuromodulation technique using the interference modulation waveform produced by several high-frequency electrical stimuli to treat neurodegenerative diseases. The device and auxiliary software constitute a non-invasive neuromodulation system. The technical problems related to the multichannel high-precision output of the device were solved by an analog phase accumulator and a special driving circuit to reduce crosstalk. The function of measuring bioimpedance in real time was integrated into the stimulator to improve effectiveness. Finite element simulation and phantom measurements were performed to find the functional relations among the target coordinates, current ratio, and electrode position in the simplified model. Then, an appropriate approach was proposed to find electrode configurations for desired target locations in a detailed and realistic mouse model. A mouse validation experiment was carried out under the guidance of a simulation, and the reliability and positioning accuracy of temporally interfering electric stimulators were verified. Stimulator improvement and precision positioning solutions promise opportunities for further studies of temporally interfering electrical stimulation.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-302X112020Diverse Partitiviruses From the Phytopathogenic Fungus,Rosellinia necatrix1064 ENPaulTelengechInstitute of Plant Science and Resources, Okayama UniversitySakaeHisanoInstitute of Plant Science and Resources, Okayama UniversityCyrusMugambiInstitute of Plant Science and Resources, Okayama UniversityKiwamuHyodoInstitute of Plant Science and Resources, Okayama UniversityJuan ManuelArjona-LopezInstitute of Plant Science and Resources, Okayama UniversityCarlos JoseLopez-HerreraInstitute for Sustainable Agriculture,Spanish Research CouncilSatokoKanematsuInstitute of Fruit Tree Science, National Agriculture and Food Research Organization (NARO)HidekiKondoInstitute of Plant Science and Resources, Okayama UniversityNobuhiroSuzukiInstitute of Plant Science and Resources, Okayama UniversityPartitiviruses (dsRNA viruses, familyPartitiviridae) are ubiquitously detected in plants and fungi. Although previous surveys suggested their omnipresence in the white root rot fungus,Rosellinia necatrix, only a few of them have been molecularly and biologically characterized thus far. We report the characterization of a total of 20 partitiviruses from 16R. necatrixstrains belonging to 15 new species, for which "Rosellinia necatrix partitivirus 11-Rosellinia necatrix partitivirus 25" were proposed, and 5 previously reported species. The newly identified partitiviruses have been taxonomically placed in two genera,Alphapartitivirus, andBetapartitivirus. Some partitiviruses were transfected into reference strains of the natural host,R. necatrix, and an experimental host,Cryphonectria parasitica, using purified virions. A comparative analysis of resultant transfectants revealed interesting differences and similarities between the RNA accumulation and symptom induction patterns ofR. necatrixandC. parasitica. Other interesting findings include the identification of a probable reassortment event and a quintuple partitivirus infection of a single fungal strain. These combined results provide a foundation for further studies aimed at elucidating mechanisms that underly the differences observed.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-665X102022Does ESG investment reduce carbon emissions in China?977049ENYingnanCongBusiness School, China University of Political Science and LawChenZhuBusiness School, China University of Political Science and LawYufeiHouBusiness School, China University of Political Science and LawShuairuTianResearch Center of Finance, Shanghai Business SchoolXiaojingCaiGraduate School of Humanities and Social Sciences, Okayama UniversityThis study explores the relationship between ESG investments and carbon emissions in China. Our results show that 1% increase in environmental investments would cause 0.246% decrease in CO2 emissions and 0.558% decrease in carbon emission intensity. The impact of ESG investment is heterogeneous across the developed and underdeveloped regions. Environmental investments in the advanced eastern region have significantly improved carbon productivity. In contrast, environmental investments in the central and western regions significantly reduced carbon emissions, but they have little impact on carbon productivity.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224142023E3-ubiquitin ligases and recent progress in osteoimmunology1120710ENYosukeAsanoDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical SciencesYoshinoriMatsumotoDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical SciencesJunWadaDepartment of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical SciencesRobertRottapelPrincess Margaret Cancer Center, University Health Network, University of TorontoUbiquitin-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.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1663-4365122020Enhancing Working Memory Based on Mismatch Negativity Neurofeedback in Subjective Cognitive Decline Patients: A Preliminary Study263ENGuangyingPeiSchool of Life Science, Beijing Institute of TechnologyRuoshuiYangSchool of Mechatronical Engineering, Beijing Institute of TechnologyZhongyanShiSchool of Life Science, Beijing Institute of TechnologyGuoxinGuoSchool of Life Science, Beijing Institute of TechnologyShujieWangSchool of Life Science, Beijing Institute of TechnologyMiaomiaoLiuGraduate School of Natural Science and Technology, Okayama UniversityYuxiangQiuSchool of Life Science, Beijing Institute of TechnologyJinglongWuFaculty of Engineering, Okayama UniversityRitsuGoSchool of Mechatronical Engineering, Beijing Institute of TechnologyYinHanDepartment of Neurology, Xuanwu Hospital, Capital Medical UniversityTianyiYanSchool of Life Science, Beijing Institute of TechnologyMismatch negativity (MMN) is suitable for studies of preattentive auditory discriminability and the auditory memory trace. Subjective cognitive decline (SCD) is an ideal target for early therapeutic intervention because SCD occurs at preclinical stages many years before the onset of Alzheimer's disease (AD). According to a novel lifespan-based model of dementia risk, hearing loss is considered the greatest potentially modifiable risk factor of dementia among nine health and lifestyle factors, and hearing impairment is associated with cognitive decline. Therefore, we propose a neurofeedback training based on MMN, which is an objective index of auditory discriminability, to regulate sensory ability and memory as a non-pharmacological intervention (NPI) in SCD patients. Seventeen subjects meeting the standardized clinical evaluations for SCD received neurofeedback training. The auditory frequency discrimination test, the visual digital N-back (1-, 2-, and 3-back), auditory digital N-back (1-, 2-, and 3-back), and auditory tone N-back (1-, 2-, and 3-back) tasks were used pre- and post-training in all SCD patients. The intervention schedule comprised five 60-min training sessions over 2 weeks. The results indicate that the subjects who received neurofeedback training had successfully improved the amplitude of MMN at the parietal electrode (Pz). A slight decrease in the threshold of auditory frequency discrimination was observed after neurofeedback training. Notably, after neurofeedback training, the working memory (WM) performance was significantly enhanced in the auditory tone 3-back test. Moreover, improvements in the accuracy of all WM tests relative to the baseline were observed, although the changes were not significant. To the best of our knowledge, our preliminary study is the first to investigate the effects of MMN neurofeedback training on WM in SCD patients, and our results suggest that MMN neurofeedback may represent an effective treatment for intervention in SCD patients and the elderly with aging memory decline.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-302X142023Etiology of recurrent cystitis in postmenopausal women based on vaginal microbiota and the role of Lactobacillus vaginal suppository1187479ENTakanoriSekitoDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceKoichiroWadaDepartment of Urology, Shimane University Faculty of MedicineAyanoIshiiDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceTakehiroIwataDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceTakehiroMatsubaraOkayama University Hospital Biobank, Okayama University HospitalShutaTomidaCenter for Comprehensive Genomic Medicine, Okayama University HospitalMasamiWatanabeDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceMotooArakiDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceTakuyaSadahiraDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceBackground: The vaginal microbiota can be altered by uropathogenic bacteria associated with recurrent cystitis (RC), and the vaginal administration of Lactobacillus have suggested certain effects to prevent RC. The relationship between vaginal microbiota and the development of RC has not been elucidated. We aimed to clarify the etiology of RC from vaginal microbiota and importance of vaginal Lactobacillus. <br>
Methods: Vaginal samples obtained from 39 postmenopausal women were classified into four groups: healthy controls; uncomplicated cystitis; RC; and prevention (prevented RC by Lactobacillus crispatus-containing vaginal suppositories). Principal coordinate analysis and beta-diversity analysis was used to assess 16S rRNA gene sequencing data from the vaginal microbiome. <br>
Results: Cluster analysis divided the vaginal bacterial communities among 129 vaginal samples into three clusters (A, B, and C). Fourteen of 14 (100%) samples from the RC group and 51 of 53 (96%) samples from the prevention group were in clusters B and C, while 29 of 38 (76%) samples from the healthy group and 14 of 24 (58%) samples from the uncomplicated cystitis group were in cluster A. The principal coordinate analysis showed that plots in the uncomplicated cystitis group were similar to the healthy group, indicating a large separation between the RC group and the uncomplicated cystitis group. On beta-diversity analysis, there were significant differences between the healthy group and the uncomplicated cystitis group (p = 0.045), and between the RC group and the uncomplicated cystitis group or the healthy group (p = 0.001, p = 0.001, respectively). There were no significant differences between the RC group and the prevention group (p = 0.446). The top six taxa were as follows: Prevotella, Lactobacillus, Streptococcus, Enterobacteriaceae, Anaerococcus, and Bifidobacterium. Among patients with RC, Lactobacillus was undetectable before administration of suppositories, while the median relative abundance of Lactobacillus was 19% during administration of suppositories (p = 0.0211), reducing the average cystitis episodes per year (6.3 vs. 2.4, p = 0.0015). <br>
Conclusion: The vaginal microbiota of postmenopausal women with RC is differed from healthy controls and uncomplicated cystitis in terms of lack of Lactobacillus and relatively dominant of Enterobacteriaceae. Vaginal administration of Lactobacillus-containing suppositories can prevent RC by stabilizing vaginal dysbiosis and causing a loss of pathogenic bacteria virulence.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-2360112023Evaluation of the association of birth order and group childcare attendance with Kawasaki disease using data from a nationwide longitudinal survey1127053ENTakahiroNambaDepartment of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesAkihitoTakeuchiDepartment of Neonatology, National Hospital Organization Okayama Medical CenterNaomiMatsumotoDepartment of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMitsuruTsugeDepartment of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMasatoYashiroDepartment of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHirokazuTsukaharaDepartment of Pediatrics, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesTakashiYorifujiDepartment of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesBackground: Kawasaki disease (KD) is a form of pediatric systemic vasculitis. Although the etiology remains unclear, infections have been identified as possible triggers. Children with a later birth order and those who attend childcare are at a higher risk of infections due to exposure to pathogens from their older siblings and other childcare attendees. However, longitudinal studies exploring these associations are limited. Thus, we aimed to elucidate the relationship between birth order, group childcare attendance, and KD, using a nationwide longitudinal survey in Japan. <br>
Methods: In total, 36,885 children born in Japan in 2010 were included. The survey used questionnaires to identify hospitalized cases of KD. We evaluated the relationship between birth order classification, group childcare attendance, and KD prevalence every year, from 6 to 66 months of age. For each outcome, odds ratios (ORs), and 95% confidence intervals (CIs) were estimated after adjusting for child factors, parental factors, and region of residence. <br>
Results: Children with higher birth orders were more likely to be hospitalized with KD at 6-18 months of age (second child OR: 1.77, 95% CI: 1.25-2.51; third child OR: 1.70, 95% CI: 1.08-2.65). This trend was stronger for children who did not attend group childcare (second child OR: 2.51, 95% CI: 1.57-4.01; third child OR: 2.41, 95% CI: 1.30-4.43). An increased risk of KD hospitalization owing to the birth order was not observed in any age group for children in the childcare group. <br>
Conclusions: Children with higher birth orders were at high risk for hospitalization due to KD at 6-18 months of age. The effect of birth order was more prominent among the children who did not attend group childcare.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-462X142023FZL, a dynamin-like protein localized to curved grana edges, is required for efficient photosynthetic electron transfer in Arabidopsis1279699ENYuOgawaInstitute of Plant Science and Resources, Okayama UniversityMegumiIwanoGraduate School of Biostudies, Kyoto UniversityToshiharuShikanaiDepartment of Botany, Graduate School of Science, Kyoto UniversityWataruSakamotoInstitute of Plant Science and Resources, Okayama UniversityPhotosynthetic electron transfer and its regulation processes take place on thylakoid membranes, and the thylakoid of vascular plants exhibits particularly intricate structure consisting of stacked grana and flat stroma lamellae. It is known that several membrane remodeling proteins contribute to maintain the thylakoid structure, and one putative example is FUZZY ONION LIKE (FZL). In this study, we re-evaluated the controversial function of FZL in thylakoid membrane remodeling and in photosynthesis. We investigated the sub-membrane localization of FZL and found that it is enriched on curved grana edges of thylakoid membranes, consistent with the previously proposed model that FZL mediates fusion of grana and stroma lamellae at the interfaces. The mature fzl thylakoid morphology characterized with the staggered and less connected grana seems to agree with this model as well. In the photosynthetic analysis, the fzl knockout mutants in Arabidopsis displayed reduced electron flow, likely resulting in higher oxidative levels of Photosystem I (PSI) and smaller proton motive force (pmf). However, nonphotochemical quenching (NPQ) of chlorophyll fluorescence was excessively enhanced considering the pmf levels in fzl, and we found that introducing kea3-1 mutation, lowering pH in thylakoid lumen, synergistically reinforced the photosynthetic disorder in the fzl mutant background. We also showed that state transitions normally occurred in fzl, and that they were not involved in the photosynthetic disorders in fzl. We discuss the possible mechanisms by which the altered thylakoid morphology in fzl leads to the photosynthetic modifications.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224112020High Mobility Group Box 1 Expression in Oral Inflammation and Regeneration1461ENKeisukeYamashiroDepartment of Periodontics and Endodontics, Okayama University HospitalHidetakaIdeguchiDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHiroakiAoyagiDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesChiakiYoshihara-HirataDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesAnnaHiraiDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesRisaSuzuki-KyoshimaDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYaoZhangDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHidenoriWakeDepartment of Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceMasahiroNishiboriDepartment of Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical ScienceTadashiYamamotoDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesShogoTakashibaDepartment of Pathophysiology-Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHigh mobility group box 1 (HMGB1) is a non-histone DNA-binding protein of about 30 kDa. It is released from a variety of cells into the extracellular milieu in response to inflammatory stimuli and acts on specific cell-surface receptors, such as receptors for advanced glycation end-products (RAGE), Toll-like receptor (TLR)2, TLR4, with or without forming a complex with other molecules. HMGB1 mediates various mechanisms such as inflammation, cell migration, proliferation, and differentiation. On the other hand, HMGB1 enhances chemotaxis acting through the C-X-C motif chemokine ligand (CXCL)12/C-X-C chemokine receptor (CXCR)4 axis and is involved in regeneration. In the oral cavity, high levels of HMGB1 have been detected in the gingival tissue from periodontitis and peri-implantitis patients, and it has been shown that secreted HMGB1 induces pro-inflammatory cytokine expression, such as interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha, which prolong inflammation. In contrast, wound healing after tooth extraction or titanium dental implant osseointegration requires an initial acute inflammation, which is regulated by secreted HMGB1. This indicates that secreted HMGB1 regulates angiogenesis and bone remodeling by osteoclast and osteoblast activation and promotes bone healing in oral tissue repair. Therefore, HMGB1 can prolong inflammation in the periodontal tissue and, conversely, can regenerate or repair damaged tissues in the oral cavity. In this review, we highlight the role of HMGB1 in the oral cavity by comparing its function and regulation with its function in other diseases. We also discuss the necessity for further studies in this field to provide more specific scientific evidence for dentistry.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-1078142023Home literacy environment and early reading skills in Japanese Hiragana and Kanji during the transition from kindergarten to primary school1052216ENTakayukiTanjiOkayama UniversityTomohiroInoueThe Chinese University of Hong KongWe examined the reciprocal associations between home literacy environment (HLE) and children's early reading skills in syllabic Hiragana and morphographic Kanji in a sample of Japanese parent-child dyads. Eighty-three children were followed from kindergarten to Grade 3 and tested on Hiragana reading accuracy in kindergarten, Hiragana word reading fluency in kindergarten and Grade 1, and Kanji reading accuracy in Grade 1 to Grade 3. Their parents answered a questionnaire about HLE [parent teaching (PT) in Hiragana and Kanji, shared book reading (SBR), and access to literacy resources (ALR)], parents' needs for early literacy support by teachers, parents' expectations for children's reading skills, parents' worry about children's homework, and mother's education level. Results showed first that ALR, but not PT and SBR, was associated with reading skills in Hiragana and Kanji. Second, whereas Hiragana reading in kindergarten was not associated with PT in Hiragana in kindergarten, it negatively predicted PT in Hiragana in Grade 1. However, Kanji reading accuracy was not associated with PT in Kanji across Grades 1 to 3. Third, parents' worry was negatively associated with children's reading performance across Grades 1 to 3 but positively associated with PT in Hiragana and Kanji. Finally, while parents' expectations were positively associated with children's reading performance across Grades 1 to 3, they were negatively associated with PT in Hiragana and Kanji in Grades 1 and 2. These results suggest that Japanese parents may be sensitive to both their children's reading performance and social expectations for school achievement and adjust their involvement accordingly during the transition period from kindergarten to early primary grades. ALR may be associated with early reading development in both Hiragana and Kanji.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224112020Identification and Modification ofPorphyromonas gingivalisCysteine Protease, Gingipain, Ideal for Screening Periodontitis1017ENKimitoHiraiDepartment of Pathophysiology—Periodontal Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTomokoYamaguchi-TomikawaDepartment of Pathophysiology—Periodontal Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityToruEguchiR&D, Sunstar Inc.HiroshiMaedaDepartment of Endodontology, Osaka Dental UniversityShogoTakashibaDepartment of Pathophysiology—Periodontal Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityChronic periodontitis is an inflammatory disease caused by the formation of oral microbial biofilms. Periodontitis is associated with general health and not only oral diseases.Porphyromonas gingivalisis a well-known keystone pathogen for periodontitis and is associated with several systemic diseases, such as diabetes mellitus and Alzheimer's disease. We previously developed a system for screening periodontitis usingP. gingivalis-specific serum immunoglobulin G (IgG) in an enzyme-linked immunosorbent assay with a sensitivity of 0.774 and a specificity of 0.586 and an area under the receiver operating characteristic curve of 0.708. However, the antigens elicited non-specific responses, since they were obtained from whole extracts of sonicated cultured bacteria. The purpose of this study was to identify antigens ideal for a sensitive and specific serum test. We identified the specific antigens using immunoaffinity columns immobilized with IgG antibodies from periodontitis patients. Liquid chromatography-tandem mass spectrometry identified 29 antigens from the elutes. Recombinant proteins for these candidates were synthesized using the wheat germ cell-free translation system and screened by dot blot analysis with serum from the columns. Three of the 16 candidates that reacted showed strongest affinities upon dot blot analysis; they included outer membrane protein 28, cysteine proteases, lysine gingipain Kgp, and arginine gingipain RgpA. Outer membrane protein 28 was not suitable for screeningP. gingivalisinfection because of its high false-negative rates. Kgp and RgpA were unstable antigens since they underwent self-digestion. They were made stable by substituting the active cysteine residues in Kgp and RgpA with alanine using site-directed mutagenesis. Using the modified antigens, we demonstrated that the patient serum IgG level against RgpA was the highest among all the antigens expressed inP. gingivalis. Moreover, the N-terminus of recombinant RgpA was excellent in differentiating between diseased and non-diseased states (with sensitivity of 0.85, specificity of 0.9, and area under the curve of 0.915). Although dot blot analysis was the only experiment used, the N-terminus of RgpA is an excellent antigen to immunologically test forP. gingivalisinfection, especially for estimating the risks for periodontitis-associated systemic diseases. In conclusion, we have developed aP. gingivalisantigen for screening periodontitis.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2234-943X132023Impact of cancer-associated fibroblasts on survival of patients with ampullary carcinoma1072106ENKoseiTakagiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesKazuhiroNomaDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesYasuoNagaiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesSatoruKikuchiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesYuzoUmedaDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesRyuichiYoshidaDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesTomokazuFujiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesKazuyaYasuiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesTakehiroTanakaDepartment of Pathology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesHajimeKashimaDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesTakahitoYagiDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesToshiyoshiFujiwaraDepartment of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesBackground: Cancer-associated fibroblasts (CAFs) reportedly enhance the progression of gastrointestinal surgery; however, the role of CAFs in ampullary carcinomas remains poorly examined. This study aimed to investigate the effect of CAFs on the survival of patients with ampullary carcinoma. <br>
Materials and methods: A retrospective analysis of 67 patients who underwent pancreatoduodenectomy between January 2000 and December 2021 was performed. CAFs were defined as spindle-shaped cells that expressed alpha-smooth muscle actin (alpha-SMA) and fibroblast activation protein (FAP). The impact of CAFs on survival, including recurrence-free (RFS) and disease-specific survival (DSS), as well as prognostic factors associated with survival, was analyzed. <br>
Results: The high-alpha-SMA group had significantly worse 5-year RFS (47.6% vs. 82.2%, p = 0.003) and 5-year DSS (67.5% vs. 93.3%, p = 0.01) than the low-alpha-SMA group. RFS (p = 0.04) and DSS (p = 0.02) in the high-FAP group were significantly worse than those in the low-FAP group. Multivariable analyses found that high alpha-SMA expression was an independent predictor of RFS [hazard ratio (HR): 3.68; 95% confidence intervals (CI): 1.21-12.4; p = 0.02] and DSS (HR: 8.54; 95% CI: 1.21-170; p = 0.03). <br>
Conclusions: CAFs, particularly alpha-SMA, can be useful predictors of survival in patients undergoing radical resection for ampullary carcinomas.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224142023Impact of oral microbiota on pathophysiology of GVHD1132983ENAkiraYamamotoDepartment of Hematology and Oncology, Okayama University HospitalYuiKambaraDepartment of Hematology and Oncology and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHideakiFujiwaraDepartment of Hematology and Oncology, Okayama University HospitalAllogeneic transplantation of hematopoietic cells is the only curative therapy for several hematopoietic disease in which patients receive cytotoxic conditioning regimens followed by infusion of hematopoietic stem cells. Although the outcomes have improved over the past decades, graft-versus-host-disease (GVHD), the most common life-threatening complication, remains a major cause of non-relapse morbidity and mortality. Pathophysiology of acute GVHD characterized by host antigen-presenting cells after tissue damage and donor T-cells is well studied, and additionally the importance of recipient microbiota in the intestine is elucidated in the GVHD setting. Oral microbiota is the second most abundant bacterial flora in the body after the intestinal tract, and it is related to chronic inflammation and carcinogenesis. Recently, composition of the oral microbiome in GVHD related to transplantation has been characterized and several common patterns, dysbiosis and enrichment of the specific bacterial groups, have been reported. This review focuses on the role of the oral microbiota in the context of GVHD.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-634X82020In vitroNeo-Genesis of Tendon/Ligament-Like Tissue by Combination of Mohawk and a Three-Dimensional Cyclic Mechanical Stretch Culture System307ENKensukeKataokaDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityRyotaKurimotoDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityHirokiTsutsumiDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityTomokiChibaDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityTomomiKatoDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityKanaShishidoDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityMarikoKatoDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityYoshiakiItoDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityYuichiroChoAnatomy and Physiological Science, Tokyo Medical and Dental UniversityOsamuHoshiAnatomy and Physiological Science, Tokyo Medical and Dental UniversityAyakoMimataResearch Core, Tokyo Medical and Dental UniversityYurikoSakamakiResearch Core, Tokyo Medical and Dental UniversityRyoNakamichiDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityMartin K.LotzDepartment of Molecular Medicine, The Scripps Research InstituteKeijiNaruseGraduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityHiroshiAsaharaDepartment of Systems BioMedicine, Tokyo Medical and Dental UniversityTendons and ligaments are pivotal connective tissues that tightly connect muscle and bone. In this study, we developed a novel approach to generate tendon/ligament-like tissues with a hierarchical structure, by introducing the tendon/ligament-specific transcription factor Mohawk (MKX) into the mesenchymal stem cell (MSC) line C3H10T1/2 cells, and by applying an improved three-dimensional (3D) cyclic mechanical stretch culture system. In our developed protocol, a combination of stableMkxexpression and cyclic mechanical stretch synergistically affects the structural tendon/ligament-like tissue generation and tendon related gene expression. In a histological analysis of these tendon/ligament-like tissues, an organized extracellular matrix (ECM), containing collagen type III and elastin, was observed. Moreover, we confirmed thatMkxexpression and cyclic mechanical stretch, induced the alignment of structural collagen fibril bundles that were deposited in a fibripositor-like manner during the generation of our tendon/ligament-like tissues. Our findings provide new insights for the tendon/ligament biomaterial fields.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2297-055X102023In vivo tracking transplanted cardiomyocytes derived from human induced pluripotent stem cells using nuclear medicine imaging1261330ENYukihiroSaitoDepartment of Cardiovascular Medicine, Okayama University HospitalNaokoNoseMolecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityToshihiroIidaDepartment of Cardiovascular Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityKaoruAkazawaDepartment of Cardiovascular Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTakayukiKannoMolecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityYukiFujimotoMolecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTakanoriSasakiOkayama Medical Innovation Center, Faculty of Medicine, Dentistry and Pharmaceutical SciencesMasaruAkehiOkayama Medical Innovation Center, Faculty of Medicine, Dentistry and Pharmaceutical SciencesTakahiroHiguchiMolecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversitySatoshiAkagiDepartment of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityMasashiYoshidaDepartment of Chronic Kidney Disease and Cardiovascular Disease, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityToruMiyoshiDepartment of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityHiroshiItoDepartment of General Internal Medicine 3, Kawasaki Medical SchoolKazufumiNakamuraDepartment of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityIntroduction: Transplantation of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) is a promising treatment for heart failure. Information on long-term cell engraftment after transplantation is clinically important. However, clinically applicable evaluation methods have not yet been established.<br>
Methods: In this study, to noninvasively assess transplanted cell engraftment, human SLC5A5, which encodes a sodium/iodide symporter (NIS) that transports radioactive tracers such as 125I, 18F-tetrafluoroborate (TFB), and 99mTc-pertechnetate (99mTcO4−), was transduced into human induced pluripotent stem cells (iPSCs), and nuclear medicine imaging was used to track engrafted human iPSC-CMs.<br>
Results: To evaluate the pluripotency of NIS-expressing human iPSCs, they were subcutaneously transplanted into immunodeficient rats. Teratomas were detected by 99mTcO4− single photon emission computed tomography (SPECT/CT) imaging. NIS expression and the uptake ability of 125I were maintained in purified human iPSC-CMs. NIS-expressing human iPSC-CMs transplanted into immunodeficient rats could be detected over time using 99mTcO4− SPECT/CT imaging. Unexpectedly, NIS expression affected cell proliferation of human iPSCs and iPSC-derived cells.<br>
Discussion: Such functionally designed iPSC-CMs have potential clinical applications as a noninvasive method of grafted cell evaluation, but further studies are needed to determine the effects of NIS transduction on cellular characteristics and functions.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2234-943X132023LOXL1 and LOXL4 are novel target genes of the Zn2+-bound form of ZEB1 and play a crucial role in the acceleration of invasive events in triple-negative breast cancer cells1142886ENDaisukeHirabayashiDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesKen-ichiYamamotoDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesAkihiroMaruyamaDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesNahokoTomonobuDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesRieKinoshitaDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYouyiChenDepartment of General Surgery & Bio-Bank of General Surgery, The Fourth Affiliated Hospital of Harbin Medical UniversityNi Luh Gede YoniKomalasariDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHitoshiMurataDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYumaGoharaDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesFanJiangDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesJinZhouMedical Oncology Department of Gastrointestinal Tumors, Liaoning Cancer Hospital & Institute, Cancer Hospital of the Dalian University of TechnologyI. Made WinarsaRumaFaculty of Medicine, Udayana UniversityI. WayanSumardikaFaculty of Medicine, Udayana UniversityAkiraYamauchiDepartment of Biochemistry, Kawasaki Medical SchoolFutoshiKuribayashiDepartment of Biochemistry, Kawasaki Medical SchoolShinichiToyookaDepartment of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYusukeInoueFaculty of Science and Technology, Division of Molecular Science, Gunma UniversityMasakiyoSakaguchiDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesBackground: EMT has been proposed to be a crucial early event in cancer metastasis. EMT is rigidly regulated by the action of several EMT-core transcription factors, particularly ZEB1. We previously revealed an unusual role of ZEB1 in the S100A8/A9-mediated metastasis in breast cancer cells that expressed ZEB1 at a significant level and showed that the ZEB1 was activated on the MCAM-downstream pathway upon S100A8/A9 binding. ZEB1 is well known to require Zn2+ for its activation based on the presence of several Zn-finger motifs in the transcription factor. However, how Zn2+-binding works on the pleiotropic role of ZEB1 through cancer progression has not been fully elucidated. <br>
Methods: We established the engineered cells, MDA-MB-231 MutZEB1 (MDA-MutZEB1), that stably express MutZEB1 (Delta Zn). The cells were then evaluated in vitro for their invasion activities. Finally, an RNA-Seq analysis was performed to compare the gene alteration profiles of the established cells comprehensively. <br>
Results: MDA-MutZEB1 showed a significant loss of the EMT, ultimately stalling the invasion. Inclusive analysis of the transcription changes after the expression of MutZEB1 (Delta Zn) in MDA-MB-231 cells revealed the significant downregulation of LOX family genes, which are known to play a critical role in cancer metastasis. We found that LOXL1 and LOXL4 remarkably enhanced cancer invasiveness among the LOX family genes with altered expression. <br>
Conclusions: These findings indicate that ZEB1 potentiates Zn2+-mediated transcription of plural EMT-relevant factors, including LOXL1 and LOXL4, whose upregulation plays a critical role in the invasive dissemination of breast cancer cells.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1663-4365122020Long-Term Continuous Cervical Spinal Cord Stimulation Exerts Neuroprotective Effects in Experimental Parkinson's Disease164ENKenKuwaharaDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTatsuyaSasakiDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTakaoYasuharaDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityMasahiroKamedaDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityYosukeOkazakiDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityKakeruHosomotoDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityIttetsuKinDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityMihokoOkazakiDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversitySatoruYabunoDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversitySatoshiKawauchiDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityYousukeTomitaDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityMichiariUmakoshiDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityKyoheiKinDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityJunMorimotoDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityJea-YoungLeeDepartment of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South FloridaNaokiTajiriDepartment of Neurophysiology and Brain Science, Graduate School of Medical Sciences, Nagoya City UniversityCesar V.BorlonganDepartment of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South FloridaIsaoDateDepartment of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityBackground: Spinal cord stimulation (SCS) exerts neuroprotective effects in animal models of Parkinson’s disease (PD). Conventional stimulation techniques entail limited stimulation time and restricted movement of animals, warranting the need for optimizing the SCS regimen to address the progressive nature of the disease and to improve its clinical translation to PD patients.</br>
Objective: Recognizing the limitations of conventional stimulation, we now investigated the effects of continuous SCS in freely moving parkinsonian rats.</br>
Methods: We developed a small device that could deliver continuous SCS. At the start of the experiment, thirty female Sprague-Dawley rats received the dopamine (DA)-depleting neurotoxin, 6-hydroxydopamine, into the right striatum. The SCS device was fixed below the shoulder area of the back of the animal, and a line from this device was passed under the skin to an electrode that was then implanted epidurally over the dorsal column. The rats were divided into three groups: control, 8-h stimulation, and 24-h stimulation, and behaviorally tested then euthanized for immunohistochemical analysis.</br>
Results: The 8- and 24-h stimulation groups displayed significant behavioral improvement compared to the control group. Both SCS-stimulated groups exhibited significantly preserved tyrosine hydroxylase (TH)-positive fibers and neurons in the striatum and substantia nigra pars compacta (SNc), respectively, compared to the control group. Notably, the 24-h stimulation group showed significantly pronounced preservation of the striatal TH-positive fibers compared to the 8-h stimulation group. Moreover, the 24-h group demonstrated significantly reduced number of microglia in the striatum and SNc and increased laminin-positive area of the cerebral cortex compared to the control group.</br>
Conclusions: This study demonstrated the behavioral and histological benefits of continuous SCS in a time-dependent manner in freely moving PD animals, possibly mediated by anti-inflammatory and angiogenic mechanisms.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2234-943X132023Lysyl oxidase-like 4 exerts an atypical role in breast cancer progression that is dependent on the enzymatic activity that targets the cell-surface annexin A21142907ENNi Luh Gede YoniKomalasariDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesNahokoTomonobuDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesRieKinoshitaDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYouyiChenDepartment of General Surgery & Bio-Bank of General Surgery, TheFourth Affiliated Hospital of Harbin Medical UniversityYoshihikoSakaguchiDepartment of Microbiology, Kitasato University School of MedicineYumaGoharaDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesFanJiangDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesKen-IchYamamotoDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHitoshiMurataDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesI Made WinarsaRumaFaculty of Medicine, Udayana UniversityI WayanSumardikaFaculty of Medicine, Udayana UniversityJinZhouMedical Oncology Department of Gastrointestinal Tumors, Liaoning Cancer Hospital & Institute, Cancer Hospital of Dalian University of TechnologyAkiraYamauchiDepartment of Biochemistry, Kawasaki Medical SchoolFutoshiKuribayashiDepartment of Biochemistry, Kawasaki Medical SchoolYusukeInoueFaculty of Science and Technology, Division of Molecular Science, Gunma UniversityShinichiToyookaDepartment of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMasakiyoSakaguchiDepartment of Cell Biology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesBackground: LOX family members are reported to play pivotal roles in cancer. Unlike their enzymatic activities in collagen cross-linking, their precise cancer functions are unclear. We revealed that LOXL4 is highly upregulated in breast cancer cells, and we thus sought to define an unidentified role of LOXL4 in breast cancer.<br>
Methods: We established the MDA-MB-231 sublines MDA-MB-231-LOXL4 mutCA and -LOXL4 KO, which stably overexpress mutant LOXL4 that loses its catalytic activity and genetically ablates the intrinsic LOXL4 gene, respectively. In vitro and in vivo evaluations of these cells’ activities of cancer outgrowth were conducted by cell-based assays in cultures and an orthotopic xenograft model, respectively. The new target (s) of LOXL4 were explored by the MS/MS analytic approach.<br>
Results: Our in vitro results revealed that both the overexpression of mutCA and the KO of LOXL4 in cells resulted in a marked reduction of cell growth and invasion. Interestingly, the lowered cellular activities observed in the engineered cells were also reflected in the mouse model. We identified a novel binding partner of LOXL4, i.e., annexin A2. LOXL4 catalyzes cell surface annexin A2 to achieve a cross-linked multimerization of annexin A2, which in turn prevents the internalization of integrin β-1, resulting in the locking of integrin β-1 on the cell surface. These events enhance the promotion of cancer cell outgrowth.<br>
Conclusions: LOXL4 has a new role in breast cancer progression that occurs via an interaction with annexin A2 and integrin β-1 on the cell surface.<br>No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-302X112020Molecular Characterization of a Novel Polymycovirus From Penicillium janthinellum With a Focus on Its Genome-Associated PASrp592789ENYukiyoSatoInstitute of Plant Science and Resources, Okayama UniversityAtifJamalCrop Diseases Research Institute, National Agricultural Research CentreHidekiKondoInstitute of Plant Science and Resources, Okayama UniversityNobuhiroSuzukiInstitute of Plant Science and Resources, Okayama UniversityThe genus Polymycovirus of the family Polymycoviridae accommodates fungal RNA viruses with different genomic segment numbers (four, five, or eight). It is suggested that four members form no true capsids and one forms filamentous virus particles enclosing double-stranded RNA (dsRNA). In both cases, viral dsRNA is associated with a viral protein termed "proline-alanine-serine-rich protein" (PASrp). These forms are assumed to be the infectious entity. However, the detailed molecular characteristics of PASrps remain unclear. Here, we identified a novel five-segmented polymycovirus, Penicillium janthinellum polymycovirus 1 (PjPmV1), and characterized its purified fraction form in detail. The PjPmV1 had five dsRNA segments associated with PASrp. Density gradient ultracentrifugation of the PASrp-associated PjPmV1 dsRNA revealed its uneven structure and a broad fractionation profile distinct from that of typical encapsidated viruses. Moreover, PjPmV1-PASrp interacted in vitro with various nucleic acids in a sequence-non-specific manner. These PjPmV1 features are discussed in view of the diversification of genomic segment numbers of the genus Polymycovirus.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-462X112020Molecular Mechanism Underlying Derepressed Male Production in Hexaploid Persimmon567249 ENKanaeMasudaGraduate School of Environmental and Life Science, Okayama UniversityNaokoFujitaGraduate School of Environmental and Life Science, Okayama UniversityHo-WenYangDepartment of Crop Sciences, University of Illinois at Urbana-ChampaignKoichiroUshijimaGraduate School of Environmental and Life Science, Okayama UniversityYasutakaKuboGraduate School of Environmental and Life Science, Okayama UniversityRyutaroTaoGraduate School of Agriculture, Kyoto UniversityTakashiAkagiGraduate School of Environmental and Life Science, Okayama UniversitySex expression in plants is often flexible and contributes to the maintenance of genetic diversity within a species. In diploid persimmons (the genus Diospyros), the sexuality is controlled by the Y chromosome-encoded small-RNA gene, OGI, and its autosomal counterpart, MeGI. Hexaploid Oriental persimmon (Diospyros kaki) evolved more flexible sex expression, where genetically male individuals carrying OGI can produce both male and female flowers (monoecy). This is due to (semi-)inactivation of OGI by the Kali-SINE retrotransposon insertion on the promoter region and the resultant DNA methylations. Instead, flower sex determination in Oriental persimmon is also dependent on DNA methylation states of MeGI. Here, we focused on a cultivar, Kumemaru, which shows stable male flower production. Our results demonstrated that cv. Kumemaru carries OGI with Kali-SINE, which was highly methylated as well as in other monoecious cultivars; nevertheless, OGI gene could have a basal expression level. Transcriptomic analysis between cv. Kumemaru and 14 cultivars that predominantly produce female flowers showed differentially expressed genes (DEGs) specific to cv. Kumemaru, which is mainly involved in stress responses. Co-expression gene networks focusing on the DEGs also suggested the involvement of stress signals, mainly via gibberellin (GA), salicylic acid (SA), and especially jasmonic acid (JA) signal pathways. We also identified potential regulators of this co-expression module, represented by the TCP4 transcription factor. Furthermore, we attempted to identify cv. Kumemaru-specific transcript polymorphisms potentially contributing to derepressed OGI expression by cataloging subsequences (k-mers) in the transcriptomic reads from cv. Kumemaru and the other 14 female cultivars. Overall, although the direct genetic factor to activate OGI remains to be solved, our results implied the involvement of stress signals in the release of silenced OGI and the resultant continuous male production.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-634X112023Mutagenic analysis of actin reveals the mechanism of His161 flipping that triggers ATP hydrolysis1105460ENMitsusadaIwasaGraduate School of Informatics, Nagoya UniversityShuichiTakedaResearch Institute for Interdisciplinary Science (RIIS), Okayama UniversityAkihiroNaritaStructural Biology Research Center, Graduate School of Science, Nagoya UniversityYuichiroMaedaResearch Institute for Interdisciplinary Science (RIIS), Okayama UniversityToshiroOdaFaculty of Health and Welfare, Tokai Gakuin UniversityThe dynamic assembly of actin is controlled by the hydrolysis of ATP, bound to the center of the molecule. Upon polymerization, actin undergoes a conformational change from the monomeric G-form to the fibrous F-form, which is associated with the flipping of the side chain of His161 toward ATP. His161 flipping from the gauche-minus to gauche-plus conformation leads to a rearrangement of the active site water molecules, including ATP attacking water (W1), into an orientation capable of hydrolysis. We previously showed that by using a human cardiac muscle a-actin expression system, mutations in the Pro-rich loop residues (A108G and P109A) and in a residue that was hydrogen-bonded to W1 (Q137A) affect the rate of polymerization and ATP hydrolysis. Here, we report the crystal structures of the three mutant actins bound to AMPPNP or ADP-P-i determined at a resolution of 1.35-1.55( )angstrom, which are stabilized in the F-form conformation with the aid of the fragmin F1 domain. In A108G, His161 remained non-flipped despite the global actin conformation adopting the F-form, demonstrating that the side chain of His161 is flipped to avoid a steric clash with the methyl group of A108. Because of the non-flipped His161, W1 was located away from ATP, similar to G-actin, which was accompanied by incomplete hydrolysis. In P109A, the absence of the bulky proline ring allowed His161 to be positioned near the Pro-rich loop, with a minor influence on ATPase activity. In Q137A, two water molecules replaced the side-chain oxygen and nitrogen of Gln137 almost exactly at their positions; consequently, the active site structure, including the W1 position, is essentially conserved. This seemingly contradictory observation to the reported low ATPase activity of the Q137A filament could be attributed to a high fluctuation of the active site water. Together, our results suggest that the elaborate structural design of the active site residues ensures the precise control of the ATPase activity of actin.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-462X102020Novel Insights Into N-Glycan Fucosylation and Core Xylosylation in C. reinhardtii1686ENAnneOltmannsInstitute of Plant Biology and Biotechnology, University of MünsterLaraHoepfnerInstitute of Plant Biology and Biotechnology, University of MünsterMartinScholzInstitute of Plant Biology and Biotechnology, University of MünsterKarenZinziusInstitute of Plant Biology and Biotechnology, University of MünsterStefanSchulzeInstitute of Plant Biology and Biotechnology, University of MünsterMichaelHipplerInstitute of Plant Science and Resources, Okayama UniversityChlamydomonas reinhardtii (C. reinhardtii) N-glycans carry plant typical beta 1,2-core xylose, alpha 1,3-fucose residues, as well as plant atypical terminal beta 1,4-xylose and methylated mannoses. In a recent study, XylT1A was shown to act as core xylosyltransferase, whereby its action was of importance for an inhibition of excessive Man1A dependent trimming. N-Glycans found in a XylT1A/Man1A double mutant carried core xylose residues, suggesting the existence of a second core xylosyltransferase in C. reinhardtii. To further elucidate enzymes important for N-glycosylation, novel single knockdown mutants of candidate genes involved in the N-glycosylation pathway were characterized. In addition, double, triple, and quadruple mutants affecting already known N-glycosylation pathway genes were generated. By characterizing N-glycan compositions of intact N-glycopeptides from these mutant strains by mass spectrometry, a candidate gene encoding for a second putative core xylosyltransferase (XylT1B) was identified. Additionally, the role of a putative fucosyltransferase was revealed. Mutant strains with knockdown of both xylosyltransferases and the fucosyltransferase resulted in the formation of N-glycans with strongly diminished core modifications. Thus, the mutant strains generated will pave the way for further investigations on how single N-glycan core epitopes modulate protein function in C. reinhardtii.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-646372019Organic Matter Preservation and Incipient Mineralization of Microtubules in 120 Ma Basaltic GlassUNSP 149ENMatthew R. M.IzawaInstitute for Planetary Materials, Okayama UniversityJames J.DynesCanadian Light Source, Inc., University of SaskatchewanNeil R.BanerjeeDepartment of Earth Sciences, University of Western OntarioRoberta L.FlemmingDepartment of Earth Sciences, University of Western OntarioLachlan C. W.MacLeanCanadian Light Source, Inc., University of SaskatchewanCallum J.HetheringtonDepartment of Geosciences, Texas Tech UniversitySergeiMatveevDepartment of Earth and Atmospheric Sciences, University of AlbertaGordonSouthamDepartment of Earth Sciences, University of Western OntarioHollow tubular structures in subaqueously-emplaced basaltic glass may represent trace fossils caused by microbially-mediated glass dissolution. Mineralized structures of similar morphology and spatial distribution in ancient, metamorphosed basaltic rocks have widely been interpreted as ichnofossils, possibly dating to similar to 3.5 Ga or greater. Doubts have been raised, however, regarding the biogenicity of the original hollow tubules and granules in basaltic glass. In particular, although elevated levels of biologically-important elements such as C, S, N, and P as well as organic compounds have been detected in association with these structures, a direct detection of unambiguously biogenic organic molecules has not been accomplished. In this study, we describe the direct detection of proteins associated with tubular textures in basaltic glass using synchrotron X-ray spectromicroscopy. Protein-rich organic matter is shown to be associated with the margins of hollow and partly-mineralized tubules. Furthermore, a variety of tubule-infilling secondary minerals, including Ti-rich oxide phases, were observed filling and preserving the microtextures, demonstrating a mechanism whereby cellular materials may be preserved through geologic time.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2296-9144102023Parameter search of a CPG network using a genetic algorithm for a snake robot with tactile sensors moving on a soft floor1138019ENHajimeTamuraGraduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityTetsushiKamegawaGraduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityWhen a snake robot explores a collapsed house as a rescue robot, it needs to move through various obstacles, some of which may be made of soft materials, such as mattresses. In this study, we call mattress-like environment as a soft floor, which deforms when some force is added to it. We focused on the central pattern generator (CPG) network as a control for the snake robot to propel itself on the soft floor and constructed a CPG network that feeds back contact information between the robot and the floor. A genetic algorithm was used to determine the parameters of the CPG network suitable for the soft floor. To verify the obtained parameters, comparative simulations were conducted using the parameters obtained for the soft and hard floor, and the parameters were confirmed to be appropriate for each environment. By observing the difference in snake robot's propulsion depending on the presence or absence of the tactile sensor feedback signal, we confirmed the effectiveness of the tactile sensor considered in the parameter search.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-462X112020Postharvest Properties of Ultra-Late Maturing Peach Cultivars and Their Attributions to Melting Flesh (M) Locus: Re-evaluation of M Locus in Association With Flesh Texture554158ENRyoheiNakanoExperimental Farm of Graduate School of Agriculture, Kyoto UniversityTakashiKawaiGraduate School of Environmental and Life Science, Okayama UniversityYosukeFukamatsuGraduate School of Environmental and Life Science, Okayama UniversityKagariAkitaGraduate School of Environmental and Life Science, Okayama UniversitySakineWatanabeGraduate School of Environmental and Life Science, Okayama UniversityTakahiroAsanoGraduate School of Environmental and Life Science, Okayama UniversityDaisukeTakataFaculty of Food and Agricultural Sciences, Fukushima UniversityMamoruSatoFaculty of Food and Agricultural Sciences, Fukushima UniversityFumioFukudaGraduate School of Environmental and Life Science, Okayama UniversityKoichiroUshijimaGraduate School of Environmental and Life Science, Okayama UniversityThe postharvest properties of two ultra-late maturing peach cultivars, "Tobihaku" (TH) and "Daijumitsuto" (DJ), were investigated. Fruit were harvested at commercial maturity and held at 25 degrees C. TH exhibited the characteristics of normal melting flesh (MF) peach, including rapid fruit softening associated with appropriate level of endogenous ethylene production In contrast, DJ did not soften at all during 3 weeks experimental period even though considerable ethylene production was observed. Fruit of TH and DJ were treated with 5,000 ppm of propylene, an ethylene analog, continuously for 7 days. TH softened rapidly whereas DJ maintained high flesh firmness in spite of an increase in endogenous ethylene production, suggesting that DJ but not TH lacked the ability to be softened in response to endogenous and exogenous ethylene/propylene. DNA-seq analysis showed that tandem endo-polygalacturonase (endoPG) genes located at melting flesh (M) locus, Pp-endoPGM (PGM), and Pp-endoPGF (PGF), were deleted in DJ. The endoPG genes at M locus are known to control flesh texture of peach fruit, and it was suggested that the non-softening property of DJ is due to the lack of endoPG genes. On the other hand, TH possessed an unidentified M haplotype that is involved in determination of MF phenotype. Structural identification of the unknown M haplotype, designated as M-0, through comparison with previously reported M haplotypes revealed distinct differences between PGM on M-0 haplotype (PGM-M-0) and PGM on other haplotypes (PGM-M-1). Peach M haplotypes were classified into four main haplotypes: M-0 with PGM-M-0; M-1 with both PGM-M-1 and PGF; M-2 with PGM-M-1; and M-3 lacking both PGM and PGF. Re-evaluation of M locus in association with MF/non-melting flesh (NMF) phenotypes in more than 400 accessions by using whole genome shotgun sequencing data on database and/or by PCR genotyping demonstrated that M-0 haplotype was the common haplotype in MF accessions, and M-0 and M-1 haplotypes were dominant over M-2 and M-3 haplotypes and co-dominantly determined the MF trait. It was also assumed on the basis of structural comparison of M haplotypes among Prunus species that the ancestral haplotype of M-0 diverged from those of the other haplotypes before the speciation of Prunus persica.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2235-2988122022Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation992198ENHiroshiYamadaDepartment of Neuroscience, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityTadashiAbeDepartment of Neuroscience, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityHikaruNagaokaDivision of Malaria Research, Proteo-Science Center, Ehime UniversityEizoTakashimaDivision of Malaria Research, Proteo-Science Center, Ehime UniversityRyoNittaDivision of Structural Medicine and Anatomy, Department of Physiology and Cell Biology, Kobe University Graduate School of MedicineMasahiroYamamotoDepartment of Immunoparasitology, Research Institute for Microbial Diseases, Osaka UniversityKohjiTakeiDepartment of Neuroscience, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityIrgb6 is a member of interferon gamma-induced immunity related GTPase (IRG), and one of twenty "effector" IRGs, which coordinately attack parasitophorous vacuole membrane (PVM), causing death of intracellular pathogen. Although Irgb6 plays a pivotal role as a pioneer in the process of PVM disruption, the direct effect of Irgb6 on membrane remained to be elucidated. Here, we utilized artificial lipid membranes to reconstitute Irgb6-membrane interaction in vitro, and revealed that Irgb6 directly deformed the membranes. Liposomes incubated with recombinant Irgb6 were drastically deformed generating massive tubular protrusions in the absence of guanine nucleotide, or with GMP-PNP. Liposome deformation was abolished by incubating with Irgb6-K275A/R371A, point mutations at membrane targeting residues. The membrane tubules generated by Irgb6 were mostly disappeared by the addition of GTP or GDP, which are caused by detachment of Irgb6 from membrane. Binding of Irgb6 to the membrane, which was reconstituted in vitro using lipid monolayer, was stimulated at GTP-bound state. Irgb6 GTPase activity was stimulated by the presence of liposomes more than eightfold. Irgb6 GTPase activity in the absence of membrane was also slightly stimulated, by lowering ionic strength, or by increasing protein concentration, indicating synergistic stimulation of the GTPase activity. These results suggest that membrane targeting of Irgb6 and resulting membrane deformation does not require GTP, but converting into GTP-bound state is crucial for detaching Irgb6 from the membrane, which might coincident with local membrane disruption.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224132022Responses of regulatory and effector T-cells to low-dose interleukin-2 differ depending on the immune environment after allogeneic stem cell transplantation891925ENYusukeMeguriDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesTakeruAsanoDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesTakanoriYoshiokaDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMikiIwamotoDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesShuntaroIkegawaDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHiroyukiSugiuraDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYurikoKishiDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMakotoNakamuraDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYasuhisaSandoDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesTakumiKondoDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYuichiSumiiDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYoshinobuMaedaDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesKen-IchiMatsuokaDepartment of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesCD4(+)Foxp3(+) regulatory T cells (Tregs) play a central role in the maintenance of immune tolerance after allogeneic hematopoietic stem cell transplantation (HSCT). Tregs promptly respond to low concentrations of IL-2 through the constitutive expression of high-affinity IL-2 receptors. It has been reported that low-dose IL-2 therapy increased circulating Tregs and improved clinical symptoms of chronic GVHD. Clinical studies of IL-2 therapy so far have mainly targeted patients in the chronic phase of transplantation when acute immune responses has subsided. However, the biological and clinical effects of exogenous IL-2 in an acute immune environment have not been well investigated. In the current study, we investigated the impact of exogenous IL-2 therapy on the post-transplant homeostasis of T cell subsets which influence the balance between GVHD and GVL in the acute phase, by setting the various immune environments early after HSCT in murine model. We initially found that 5,000 IU of IL-2 was enough to induce the active proliferation of Treg without influencing other conventional T cells (Tcons) when administered to normal mice. However, activated Tcons showed the response to the same dose of IL-2 in recipients after allogeneic HSCT. In a mild inflammatory environment within a threshold, exogenous IL-2 could effectively modulate Treg homeostasis with just limited influence to activated T cells, which resulted in an efficient GVHD suppression. In contrast, in a severely inflammatory environment, exogenous IL-2 enhanced activated T cells rather than Tregs, which resulted in the exacerbation of GVHD. Of interest, in an immune-tolerant state after transplant, exogenous IL-2 triggered effector T-cells to exert an anti-tumor effect with maintaining GVHD suppression. These data suggested that the responses of Tregs and effector T cells to exogenous IL-2 differ depending on the immune environment in the host, and the mutual balance of the response to IL-2 between T-cell subsets modulates GVHD and GVL after HSCT. Our findings may provide useful information in the optimization of IL-2 therapy, which may be personalized for each patient having different immune status.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2624-955342022Sea Surface Temperature and Salinity in Lombok Strait Reconstructed From Coral Sr/Ca and δ18O, 1962–2012918273ENAiGendaGraduate School of Natural Science and Technology, Okayama UniversityMinoruIkeharaCenter for Advanced Marine Core Research, Kochi UniversityAtsushiSuzukiGeological Survey of Japan, National Institute of Advanced Industrial Science and Technology (AIST)AliArmanResearch and Technology Center for Application of Isotope and Radiation, National Research and Innovation AgencyMayuriInoueGraduate School of Natural Science and Technology, Okayama UniversityCoral geochemical tracers have been used in studies of the paleoclimatology and paleoceanography of the tropics and subtropics. We measured Sr/Ca and oxygen isotope ratios (δ18O) in a coral sample collected from the southern part of Lombok Strait, a significant outlet of the Indonesian Throughflow (ITF) to the Indian Ocean, to reconstruct the historical record of sea surface temperature (SST) and seawater δ18O. Seawater δ18O can be used to approximate sea surface salinity (SSS) because it reflects the balance of evaporation and precipitation. The resulting time series reconstructed SST and SSS, covering the period 1962–2012, shows no clear trend of global warming, although the record includes a large cooling event (~4°C) during 1996–1997. Although neither SST nor SSS shows a systematic relationship with El Niño–Southern Oscillation and Indian Ocean Dipole (IOD), weak but significant correlations are found partly. In addition, the coral data show signals of major IOD and El Niño events in 1994 and 1997, respectively, although climatic trends recorded in the coral are not consistent with those found along the Java-Sumatra coast. To evaluate other influences on the ITF in Lombok Strait, we compared our coral record with coral records from sites in the Java Sea, the southern part of Makassar Strait, and Ombai Strait. During the northwest monsoon (December–January–February), variations in SST and SSS at Lombok Strait site are similar to those at the Java Sea and southern Makassar sites for the period 1962–1995, which suggests that low-salinity water from the Java Sea is carried at least to the southern part of Makassar Strait where it suppresses the ITF upstream from Lombok Strait. However, the SST and SSS records differ at the three sites during the southeast monsoon (June–July–August), indicating that surface conditions in Lombok Strait vary separately from those in the Java Sea. In the longer term, although global warming has been widely identified in the Indonesian Seas, the coral record shows no clear warming trend in the southern part of Lombok Strait, where fluctuations in the ITF may be modulating the distribution of heat in the surface waters of the western Pacific and eastern Indian Ocean.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1662-5196142021Stimulus Intervals Modulate the Balance of Brain Activity in the Human Primary Somatosensory Cortex: An ERP Study571369ENYangLiuDepartment of Psychology, Suzhou University of Science and TechnologyBoDongDepartment of Psychology, Suzhou University of Science and TechnologyJiajiaYangCognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama UniversityYoshimichiEjimaCognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama UniversityJinglongWuCognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama UniversityQiongWuCognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama UniversityMingZhangCognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama UniversityNeuronal excitation and inhibition occur in the brain at the same time, and brain activation reflects changes in the sum of excitation and inhibition. This principle has been well-established in lower-level sensory systems, including vision and touch, based on animal studies. However, it is unclear how the somatosensory system processes the balance between excitation and inhibition. In the present ERP study, we modified the traditional spatial attention paradigm by adding double stimuli presentations at short intervals (i.e., 10, 30, and 100 ms). Seventeen subjects participated in the experiment. Five types of stimulation were used in the experiment: a single stimulus (one raised pin for 40 ms), standard stimulus (eight pins for 40 ms), and double stimuli presented at intervals of 10, 30, and 100 ms. The subjects were asked to attend to a particular finger and detect whether the standard stimulus was presented to that finger. The results showed a clear attention-related ERP component in the single stimulus condition, but the suppression components associated with the three interval conditions seemed to be dominant in somatosensory areas. In particular, we found the strongest suppression effect in the ISI-30 condition (interval of 30 ms) and that the suppression and enhancement effects seemed to be counterbalanced in both the ISI-10 and ISI-100 conditions (intervals of 10 and 100 ms, respectively). This type of processing may allow humans to easily discriminate between multiple stimuli on the same body part.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama2504-284X82023Sustainable development goals in teacher education: comparing syllabi in a Japanese and a Slovenian university1215500ENKhalifatullohFiel'ardhGraduate School of Education, Okayama UniversityGregorTorkarFaculty of Education, University of LjubljanaHanaRožmanFaculty of Education, University of LjubljanaHirokiFujiiGraduate School of Education, Okayama UniversityIntroduction: This research aims to explore the integration of Sustainable Development Goals (SDGs) within teacher education programs, focusing on the Faculty of Education at Okayama University, Japan and the University of Ljubljana, Slovenia.<br>
Methods: We employed a qualitative content analysis of the syllabi (n = 2,079 from Okayama University; n = 504 from University of Ljubljana) and combined it with insights from semi-structured interviews.<br>
Results: The analysis illuminated a strong emphasis on Quality Education (SDG 4) in both institutions. However, certain SDGs, like Climate Action (SDG 13), were less represented, marking potential areas for enhancement. Differences were also identified in the distribution of SDGs-related content between compulsory and elective courses, indicating institutional priorities. Interview reflections emphasized the pivotal role of educators in realizing SDGs and highlighted the necessity of collaboration to achieve these global objectives.<br>
Discussion: The insights from interviews and syllabi content analysis underscore the urgency to bridge the identified gaps in SDG coverage. Disparities in emphasis between the two Education for Sustainable Development (ESD)-committed universities were noted, suggesting the importance of fostering strategy exchange and partnerships across institutions.<br>
Conclusion: Enhancing the alignment of teacher education programs with SDGs requires collective efforts. By addressing the gaps and promoting effective collaboration, these programs can achieve greater relevance and efficacy in promoting the SDGs.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-302X152024Tetrathionate hydrolase from the acidophilic microorganisms1338669ENTadayoshiKanaoDepartment of Agricultural and Biological Chemistry, Graduate School of Environment, Life, Natural Science, and Technology, Okayama UniversityTetrathionate hydrolase (TTH) is a unique enzyme found in acidophilic sulfur-oxidizing microorganisms, such as bacteria and archaea. This enzyme catalyzes the hydrolysis of tetrathionate to thiosulfate, elemental sulfur, and sulfate. It is also involved in dissimilatory sulfur oxidation metabolism, the S-4-intermediate pathway. TTHs have been purified and characterized from acidophilic autotrophic sulfur-oxidizing microorganisms. All purified TTHs show an optimum pH in the acidic range, suggesting that they are localized in the periplasmic space or outer membrane. In particular, the gene encoding TTH from Acidithiobacillus ferrooxidans (Af-tth) was identified and recombinantly expressed in Escherichia coli cells. TTH activity could be recovered from the recombinant inclusion bodies by acid refolding treatment for crystallization. The mechanism of tetrathionate hydrolysis was then elucidated by X-ray crystal structure analysis. Af-tth is highly expressed in tetrathionate-grown cells but not in iron-grown cells. These unique structural properties, reaction mechanisms, gene expression, and regulatory mechanisms are discussed in this review.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-2392112020Transcriptomic Analysis of the Kuruma PrawnMarsupenaeus japonicusReveals Possible Peripheral Regulation of the Ovary541ENNaoakiTsutsuiFaculty of Science, Ushimado Marine Institute, Okayama UniversityYasuhisaKobayashiFaculty of Science, Ushimado Marine Institute, Okayama UniversityKouichiIzumikawaResearch Institute for Fisheries Science, Okayama Prefectural Technology Center for Agriculture, Forestry, and FisheriesTatsuyaSakamotoFaculty of Science, Ushimado Marine Institute, Okayama UniversityCrustacean reproduction has been hypothesized to be under complex endocrinological regulation by peptide hormones. To further improve our understanding of the mechanisms underlying this complex regulation, knowledge is needed regarding the hormones not only of the central nervous system (CNS) such as the X-organ/sinus gland (XOSG), brain, and thoracic ganglia, but also the peripheral gonadal tissues. For example, in vertebrates, some gonadal peptide hormones including activin, inhibin, follistatin, and relaxin are known to be involved in the reproductive physiology. Therefore, it is highly likely that some peptide factors from the ovary are serving as the signals among peripheral tissues and central nervous tissues in crustaceans. In this work, we sought to find gonadal peptide hormones and peptide hormone receptors by analyzing the transcriptome of the ovary of the kuruma prawnMarsupenaeus japonicus. The generated ovarian transcriptome data led to the identification of five possible peptide hormones, including bursicon-alpha and -beta, the crustacean hyperglycemic hormone (CHH)-like peptide, insulin-like peptide (ILP), and neuroparsin-like peptide (NPLP). Dominant gene expressions for the bursicons were observed in the thoracic ganglia and the ovary, in the CNS for the CHH-like peptide, in the heart for NPLP, and in the ovary for ILP. Since the gene expressions of CHH-like peptide and NPLP were affected by a CHH (Penaeus japonicussinus gland peptide-I) from XOSG, we produced recombinant peptides for CHH-like peptide and NPLP usingEscherichia coliexpression system to examine their possible peripheral regulation. As a result, we found that the recombinant NPLP increased vitellogenin gene expression in incubated ovarian tissue fragments. Moreover, contigs encoding putative receptors for insulin-like androgenic gland factor, insulin, neuroparsin, and neuropeptide Y/F, as well as several contigs encoding orphan G-protein coupled receptors and receptor-type guanylyl cyclases were also identified in the ovarian transcriptome. These results suggest that reproductive physiology in crustaceans is regulated by various gonadal peptide hormones, akin to vertebrates.No potential conflict of interest relevant to this article was reported.Frontiers MediaActa Medica Okayama1664-3224142023“Input/output cytokines” in epidermal keratinocytes and the involvement in inflammatory skin diseases1239598ENShinMorizaneDepartment of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesTomoyukiMukaiDepartment of Immunology and Molecular Genetics, Kawasaki Medical SchoolKoSunagawaDepartment of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesKotaTachibanaDepartment of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYoshioKawakamiDepartment of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMamoruOuchidaDepartment of Molecular Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesConsidering the role of epidermal keratinocytes, they occupy more than 90% of the epidermis, form a physical barrier, and also function as innate immune barrier. For example, epidermal keratinocytes are capable of recognizing various cytokines and pathogen-associated molecular pattern, and producing a wide variety of inflammatory cytokines, chemokines, and antimicrobial peptides. Previous basic studies have shown that the immune response of epidermal keratinocytes has a significant impact on inflammatory skin diseases. The purpose of this review is to provide foundation of knowledge on the cytokines which are recognized or produced by epidermal keratinocytes. Since a number of biologics for skin diseases have appeared, it is necessary to fully understand the relationship between epidermal keratinocytes and the cytokines. In this review, the cytokines recognized by epidermal keratinocytes are specifically introduced as "input cytokines", and the produced cytokines as "output cytokines". Furthermore, we also refer to the existence of biologics against those input and output cytokines, and the target skin diseases. These use results demonstrate how important targeted cytokines are in real skin diseases, and enhance our understanding of the cytokines.No potential conflict of interest relevant to this article was reported.