Elsevier BV Acta Medica Okayama 1572-1000 58 2026 Pathological features of residual head and neck cancer after near-infrared photoimmunotherapy: implications of an unfavorable tumor immune microenvironment 105422 EN Takuma Makino Department of Otolaryngology - Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuharu Sato Department of Hematopathology, Okayama University Graduate School of Health Sciences Asami Nishikori Department of Hematopathology, Okayama University Graduate School of Health Sciences Yuto Naoi Department of Otolaryngology - Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mizuo Ando Department of Otolaryngology - Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Purpose: Near-infrared photoimmunotherapy has been approved in Japan for unresectable locally advanced or recurrent head and neck cancer. While this therapy theoretically induces selective necrosis of epidermal growth factor receptor-expressing tumor cells following administration of cetuximab sarotalocan sodium and irradiation with 690-nm light, residual disease is not uncommon in clinical practice. The mechanisms underlying such persistence remain poorly understood.<br> Methods: We examined two patients with head and neck squamous cell carcinoma who underwent salvage resection due to residual disease after four cycles of near-infrared photoimmunotherapy. Resected specimens were subjected to histopathological evaluation, including immunohistochemistry for epidermal growth factor receptor, cluster of differentiation 8, programmed cell death protein 1, and forkhead box P3 proteins.<br> Results: In both cases, residual tumor cells were predominantly localized in the superficial mucosal layers, while deeper layers were replaced by fibrosis. All residual tumors retained expression of epidermal growth factor receptor. Lymphocytic infiltration was sparse, with scattered cluster of differentiation 8-positive cells but few programmed cell death protein 1-positive or forkhead box P3-positive lymphocytes. These findings suggest that the tumor microenvironment surrounding residual tumors may have been unfavorable for the induction of robust antitumor immune responses.<br> Conclusion: Residual tumors after near-infrared photoimmunotherapy can occur despite adequate laser irradiation and epidermal growth factor receptor expression. Our observations imply that insufficient immune activation within the tumor microenvironment may contribute to treatment resistance. Further characterization of the post-photoimmunotherapy tumor microenvironment will be essential to better understand treatment mechanisms and to optimize therapeutic efficacy. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1755-4330 18 6 2026 Two-dimensional metal–organic frameworks offer all-in-one cocatalysts for photocatalytic overall water splitting 1053 1061 EN Jingyan Guan Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University Hajime Suzuki Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University Kazuhide Kamiya Research Center for Solar Energy Chemistry, Graduate School of Engineering Science, The University of Osaka Takashi Harada Research Center for Solar Energy Chemistry, Graduate School of Engineering Science, The University of Osaka Rintaro Adachi Graduate School of Natural Science and Technology, Okayama University Osamu Tomita Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University Hirofumi Kurokawa Institute of Multidisciplinary Research for Advanced Materials, Tohoku University Daisuke Unabara Institute of Multidisciplinary Research for Advanced Materials, Tohoku University Koji Yonekura Institute of Multidisciplinary Research for Advanced Materials, Tohoku University Naoya Fukui Research Institute for Science and Technology, Tokyo University of Science Hiroaki Maeda Research Institute for Science and Technology, Tokyo University of Science Kunihisa Sugimoto Department of Chemistry, Kindai University Yuichi Yamaguchi Department of Applied Chemistry, Faculty of Science, Tokyo University of Science Akinori Saeki Innovative Catalysis Science Division, Institute for Open and Transdisciplinary Research Initiatives, The University of Osaka Akira Yamakata Graduate School of Natural Science and Technology, Okayama University Akihiko Kudo Department of Applied Chemistry, Faculty of Science, Tokyo University of Science Ryu Abe Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University Ryota Sakamoto Department of Chemistry, Graduate School of Science, Tohoku University Photocatalytic overall water splitting holds great promise for sustainable hydrogen production. For overall water splitting performed by one-step excitation, both the hydrogen evolution and oxygen evolution reactions are hindered kinetically; hence, it is necessary to employ site-selective modification of photocatalysts with hydrogen-evolving and oxygen-evolving cocatalysts. However, critical challenges remain, including the need for cumbersome, multi-step photodeposition processes and durable blocking layers to inhibit the reverse reactions. Here we find a conductive, two-dimensional metal–organic framework to serve as a simple, multifunctional cocatalyst. We loaded this framework on SrTiO3:Al, an overall water-splitting photocatalyst, using a one-step self-assembly method. The framework cocatalyst promoted steady photocatalysis with an apparent quantum efficiency of 31.5% at 350 nm, free from the reverse reaction even without blocking layers. We proposed the operational principles for the cocatalyst activity using spectroscopic, electrochemical and theoretical analyses. This two-dimensional metal–organic framework offers an all-in-one approach for designing efficient and practical one-step excitation overall water-splitting systems. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1557-1963 21 2026 Surface Plasmon Resonances in Silver Nanodendrites : Trunk Length and Branch Connectivity Dependence 2247 2258 EN Qingyuan Ma Graduate School of Informatics and Engineering, The University of Electro-communications Yuki Kishida Graduate School of Informatics and Engineering, The University of Electro-communications Nobuyuki Takeyasu Graduate School of Natural Science and Technology, Okayama University Satoru Shoji Graduate School of Informatics and Engineering, The University of Electro-communications This study systematically investigates how trunk length and branch connectivity govern surface plasmon resonances in silver nanodendrites in the infrared (IR) region using a computational modeling strategy. We show that a continuous conductive trunk is essential for exciting long-wavelength collective plasmon modes. In a simulated bottom-up construction scheme, the trunk length is gradually increased to conductively connect additional branches to the backbone. Our results reveal that the fundamental δ mode resonance can be deterministically tuned across the mid-infrared spectrum (from 3840 nm to 4360 nm) primarily by controlling the trunk connectivity. As the number of connected branches grows, the lowest-order collective resonance peak exhibits a systematic redshift, and its resonance wavelength scales linearly with the effective dipole length Leff of the electron oscillation path. Concurrently, new higher-order modes emerge as local resonances of the connected substructures. These observations indicate that interrupting the conductive pathway causes a global collective mode to decompose into multiple resonances associated with more weakly coupled subsystems. The established linear scaling relationship provides a highly predictable design rule for this “programmable” connectivity, offering a robust platform for advanced applications such as multi-spectral infrared imaging, selective chemical sensing, and surface-enhanced infrared absorption (SEIRA) spectroscopy, where precise, a priori control over narrow-band infrared resonances is essential. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 2731-5894 2026 Comparative study of Ni–CeO2 catalysts prepared by impregnation and coprecipitation for CO2 methanation EN Nobuko Fukuda Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Shuichi Imano Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Nozomi Nakahara Graduate School of Environmental, Life, Natural Science and Technology, Okayama University This study explores how synthesis methods affect the structure and CO2 methanation performance of Ni–CeO2 catalysts prepared by coprecipitation and impregnation under identical conditions. Coprecipitation generated particles below 100 nm with uniform elemental distribution, together with large bulk-like particles exhibiting locally concentrated Ni species, attributed to differences in hydroxide solubility. Impregnation, by contrast, produced very large particles (> 500 nm) with smaller particles attached, while maintaining relatively homogeneous elemental distribution. Coprecipitated catalysts showed slightly higher surface area and oxygen vacancy concentration, resulting in higher apparent turnover frequencies (TOFapp) below 300 °C due to enhanced CO2 adsorption and high Ni site density. However, at temperatures above 350 °C, impregnated catalysts displayed higher CH4 selectivity and TOFapp, indicating reduced kinetic limitations and more efficient active-site utilization. These results provide insights for rational design of efficient CO2 methanation catalysts. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 80 2 2026 Impact of Proteinuria on Postoperative Complications Following Colorectal Cancer Surgery 131 139 EN Shunsuke Nakata Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Fumiaki Takatsu Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Yoshihiro Mikuriya Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Tomokazu Kakishita Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Shinji Hato Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Koji Ohta Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Takaya Kobatake Department of Gastroenterological Surgery, National Hospital Organization Shikoku Cancer Center Original Article 10.18926/AMO/70455 Colorectal surgery is associated with a high incidence of postoperative complications regardless of the advances in surgical techniques and multidisciplinary treatment. Proteinuria is common in patients with malignancies, but few studies have investigated the association between preoperative proteinuria and patient prognoses, especially postoperative complications. We investigated the impact of proteinuria on patients undergoing colorectal surgery in a single-center, retrospective cohort study of 767 patients who underwent surgical resection for colorectal cancer between January 2016 and December 2022 at the National Hospital Organization Shikoku Cancer Center. Among them, 81 patients with preoperative proteinuria were compared with the control group of 686 patients without proteinuria. Our analyses revealed that the patients with proteinuria had malnutrition with a significantly lower prognostic nutritional index compared to the no-proteinuria control group (p<0.001). The proteinuria group had a significantly advanced tumor stage (p=0.005), experienced more bleeding during the surgery (p=0.002), and required more transfusions (p<0.001). Postoperative complications were significantly more frequent in the proteinuria group (p=0.03), thus demonstrating that proteinuria was independently associated with postoperative complications (p=0.045). Proteinuria in patients undergoing colorectal cancer surgery can therefore be considered a risk factor for postoperative complications. No potential conflict of interest relevant to this article was reported.

MDPI AG Acta Medica Okayama 2079-6447 5 2 2026 Solution-Processable Near-Infrared-Absorbing Dye: Thiophene-Substituted N-Phenylphenothiazine Radical Cations for Stable Thin Films 14 EN Masafumi Yano Faculty of Chemistry, Material and Bioengineering, Kansai University Kengo Sakai Faculty of Chemistry, Material and Bioengineering, Kansai University Minami Ueda Faculty of Chemistry, Material and Bioengineering, Kansai University Koichi Mitsudo Division of Applied Chemistry, Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Yukiyasu Kashiwagi Osaka Research Institute of Industrial Science and Technology We report a π-extended N-phenylphenothiazine dye bearing thiophene substituents, designed to address the practical compromise between long-wavelength near-infrared (NIR) absorption and the isolability of a stable radical cation state. The target compound was synthesized via Suzuki–Miyaura cross-coupling and exhibited good solubility in common organic solvents. Cyclic voltammetry in dichloromethane showed a reversible one-electron oxidation at E0 = 0.19 V vs. Fc/Fc+. Chemical oxidation afforded the corresponding radical cation, which showed an intense NIR absorption maximum at 910 nm. DFT calculations support thiophene-induced narrowing of the HOMO–SOMO gap and predict a pronounced bathochromic shift of the main absorption band. The radical cation was isolated as a stable PF6− salt and readily processed into spin-coated films, which retained strong NIR absorption and remained stable for months under ambient conditions. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0956-7135 183 2026 Monitoring postharvest water loss in eggplants (Solanum melongena L.) using UV-induced fluorescence imaging and multivariate analysis 111902 EN Vincent Rotich Laboratory of Biosensing Engineering, Graduate School of Agriculture, Kyoto University Tianqi Gao Laboratory of Biosensing Engineering, Graduate School of Agriculture, Kyoto University Panintorn Prempree Laboratory of Biosensing Engineering, Graduate School of Agriculture, Kyoto University Takahiro Hayashi Laboratory of Biosensing Engineering, Graduate School of Agriculture, Kyoto University Kazuhiko Namba Faculty of Environmental, Life, Natural Science and Technology, Okayama University Mitsuji Monta Faculty of Environmental, Life, Natural Science and Technology, Okayama University Motomi Nishimoto Technology and Innovation Center, Daikin Industries, Ltd. Naoshi Kondo Laboratory of Biosensing Engineering, Graduate School of Agriculture, Kyoto University Eggplant (Solanum melongena L.) is susceptible to significant postharvest losses primarily due to water loss during storage, which affects market quality by causing texture and glossiness degradation. We investigated whether UV-induced fluorescence imaging and EEM (Excitation-Emission Matrix) fluorescence spectroscopy can non-destructively monitor WL under four storage regimes (10 °C/95 % RH, 20 °C/95 % RH, 20 °C/75 % RH, 10 °C/75 % RH). EEMs exhibited three regions; a 365/420 nm blue emission increased most under warm, low-humidity storage and is consistent with phenolic/lignin-related fluorescence. Side-view fluorescence (FL) images showed progressive blue-white emission and surface textural changes that tracked gravimetric water loss (WL). A PLSR model using combined color and texture features from FL and reflectance (CL) images achieved R2CV = 0.88 (RMSECV = 3.47 %) with only six features. To test a minimal predictor, we fit an Analysis of Covariance (ANCOVA) using Day-1 FL MeanBlue as a covariate and storage category as a factor with Leave One Out Cross-validation (LOOCV); this forecasted cumulative WL with R2LOOCV = 0.92 and MAE = 1.88 %. Importantly, this ANCOVA model using Day-1 blue-band fluorescence as a covariate was predictive only under 20 °C/75 % RH; under the other conditions, its contribution was weak. Linear Discriminant Analysis (LDA) and Support Vector Machine (SVM) models achieved accuracies of 94.4 % and 85.2 %, respectively, in differentiating storage conditions. These results support low-cost FL imaging as a practical tool to monitor WL and storage stress. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 80 1 2026 Changes in Prescribing Patterns of Antiviral Drugs before and after Public Coverage Termination among Hospitalized COVID-19 Patients in Regional Hospitals in Japan: A Retrospective, Multicenter Study 55 62 EN Hidemasa Akazawa Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Hideharu Hagiya Department of Infectious Diseases, Okayama University Hospital Shinnosuke Fukushima Department of Infectious Diseases, Okayama University Hospital Shohei Yamamoto Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Yasuhiro Nakano Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Fumio Otsuka Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Original Article 10.18926/AMO/70073 In Japan, antiviral agents for COVID-19 were freely available until September 2023 as part of national policy. This study evaluated changes in these agents’ prescribing patterns and the patient outcomes following the policy shift. We conducted a multicenter retrospective study at four hospitals in Japan’s Okayama and Kagawa prefectures from January 2022 to March 2024. The study period was divided into the public-expenditure phase (January 2022 to September 2023) and the post-expenditure phase (October 2023 to March 2024). We extracted the hospitalized patients’ clinical data from the electronic database. The study’s primary outcome was the antiviral prescription rate; the secondary outcome was in-hospital mortality. Among the 302 hospitalized patients (median age 85 years), 52.0% were classified as having a mild condition. Of the patients with mild conditions, 37.7% were diagnosed in outpatient settings prior to hospitalization. During the public-expenditure phase, 47.4% of the patients received antivirals as outpatients, mainly molnupiravir (80.9%). In the post-expenditure period, 80.0% of the patients were prescribed antivirals, mostly molnupiravir (91.7%). The antiviral prescription rate was significantly higher after the policy change. The overall in-hospital mortality was 15.8%, with no significant difference between the two periods (17.0% vs. 10.5%). Despite the termination of government funding, antiviral prescriptions remained frequent at community hospitals located in highly aging regions of western Japan such as Okayama and Kagawa prefectures. Mortality remains high among the elderly, highlighting the need for continued antiviral therapy and booster vaccinations. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 6 2025 COVID-19 and the Risks of Migraine and Headache: A Mendelian Randomization Study 413 419 EN Zhiyun Jiang Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University Ying Xi Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University Original Article 10.18926/AMO/69843 Several observational studies suggested that migraine headache attacks were associated with coronavirus disease 2019 (COVID-19). We investigated genetic causal links between COVID-19 phenotypes and the development of headache and migraine, including migraine with aura (MA) and migraine without aura (MO). We conducted a two-sample Mendelian randomization (MR) analysis to estimate the genetic association in European populations. The inverse-variance weighted (IVW) method was used as the main approach in the MR analyses, together with weighted median and MR-Egger methods. We also performed a series of sensitivity tests to assess the robustness of the MR results. The MR results demonstrated that COVID-19 severity, hospitalization, and susceptibility had no causal effect on the risks of headache, migraine, MA, or MO. No horizontal pleiotropy was detected, and the results were robust as supported by the sensitivity analysis findings. Our analyses identified no casual effect of COVID-19 severity, hospitalization, or susceptibility on the risks of headache or migraine in European populations. No potential conflict of interest relevant to this article was reported.

American Chemical Society (ACS) Acta Medica Okayama 2155-5435 15 14 2025 Mesoporous Oxyhalide Aggregates Exhibiting Improved Photocatalytic Activity for Visible-Light H2 Evolution and CO2 Reduction 12551 12562 EN Hiroto Ueki Department of Chemistry, School of Science, Institute of Science Tokyo Toshiya Tanaka Department of Chemistry, School of Science, Institute of Science Tokyo Shuji Anabuki Graduate School of Natural Science and Technology, Okayama University Ryuichi Nakada Department of Chemistry, School of Science, Institute of Science Tokyo Megumi Okazaki Department of Chemistry, School of Science, Institute of Science Tokyo Kenta Aihara Department of Chemistry, School of Science, Institute of Science Tokyo Masashi Hattori Materials and Structures Laboratory, Institute of Integrated Research, Institute of Science Tokyo Fumitaka Ishiwari Department of Applied Chemistry, Graduate School of Engineering, Osaka University Rie Haruki Institute of Materials Structure Science, High Energy Accelerator Research Organization Shunsuke Nozawa Institute of Materials Structure Science, High Energy Accelerator Research Organization Toshiyuki Yokoi Nanospace Catalysis Unit, Institute of Integrated Research, Institute of Science Tokyo Michikazu Hara Materials and Structures Laboratory, Institute of Integrated Research, Institute of Science Tokyo Osamu Ishitani Department of Chemistry, Graduate School of Advanced Science and Engineering, Hiroshima University Akinori Saeki Department of Applied Chemistry, Graduate School of Engineering, Osaka University Akira Yamakata Graduate School of Natural Science and Technology, Okayama University Kazuhiko Maeda Department of Chemistry, School of Science, Institute of Science Tokyo Oxyhalides are promising visible-light photocatalysts for water splitting and CO2 conversion; however, those exhibiting high activity for these reactions have rarely been reported. Here, we show that using water-soluble Ti complexes as precursors in the microwave-assisted hydrothermal synthesis of the oxyhalide photocatalyst Pb2Ti2O5.4F1.2 (PTOF) resulted in the production of nanoparticulate PTOF. The primary particle size of the synthesized PTOF ranged from several tens of nanometers to several hundreds of nanometers. Using Ti-citric acid or Ti-tartaric acid complexes as precursors, the PTOF was formed as mesoporous aggregates, compared with a bulky analogue (0.5–1 μm) prepared using a TiCl4 precursor. The PTOF prepared from Ti-citric acid complex had a particle size of 50–100 nm and showed a one-order-of-magnitude greater activity for H2 evolution from an aqueous ethylenediaminetetraacetic acid solution with the aid of a Rh cocatalyst. An apparent quantum yield (AQY) of 15.4 ± 1.0% at 420 nm, which is the highest among the reported oxyhalide photocatalysts, was achieved under optimal conditions. Although excess particle size reduction of PTOF lowered the H2 evolution activity, the PTOF with the smallest possible primary particle size of 15–30 nm, prepared from Ti-tartaric acid complex, showed the highest activity toward the selective reduction of CO2 into formate in a nonaqueous environment when combined with a binuclear Ru(II) complex. The CO2 reduction AQY was 10.4 ± 1.8% at 420 nm, a record-high value among metal-complex/semiconductor binary hybrid photocatalysts. This study highlights the importance of morphological control of oxyhalides for realizing their full potential as photocatalysts for artificial photosynthesis. No potential conflict of interest relevant to this article was reported.

American Chemical Society (ACS) Acta Medica Okayama 2574-0962 8 6 2025 Effects of Metal-Cation Doping on Photocatalytic H2 Evolution Activity of Layered Perovskite Oxynitride K2LaTa2O6N 3541 3552 EN Hideya Tsuchikado Department of Chemistry, School of Science, Institute of Science Tokyo Shuji Anabuki Graduate School of Natural Science and Technology, Okayama University Ovidiu Cretu Electron Microscopy Group, National Institute for Materials Science (NIMS) Yuki Kinoshita Department of Chemical Science and Engineering, School of Materials and Chemical Technology, Institute of Science Tokyo Masashi Hattori Institute of Integrated Research, Institute of Science Tokyo Yuta Shiroma Department of Chemistry, School of Science, Institute of Science Tokyo Dongxiao Fan Institute of Materials Structure Science High Energy Accelerator Research Organization Megumi Okazaki Department of Chemistry, School of Science, Institute of Science Tokyo Takuto Soma Department of Chemical Science and Engineering, School of Materials and Chemical Technology, Institute of Science Tokyo Fumitaka Ishiwari Department of Applied Chemistry, Graduate School of Engineering, Osaka University Shunsuke Nozawa Institute of Materials Structure Science High Energy Accelerator Research Organization Toshiyuki Yokoi Institute of Integrated Research, Institute of Science Tokyo Michikazu Hara Institute of Integrated Research, Institute of Science Tokyo Koji Kimoto Electron Microscopy Group, National Institute for Materials Science (NIMS) Akira Yamakata Graduate School of Natural Science and Technology, Okayama University Akinori Saeki Department of Applied Chemistry, Graduate School of Engineering, Osaka University Kazuhiko Maeda Department of Chemistry, School of Science, Institute of Science Tokyo Aliovalent cation doping into a heterogeneous photocatalyst affects several of its physicochemical properties, including its morphological characteristics, optical absorption behavior, and charge carrier dynamics, causing a drastic change in its photocatalytic activity. In the present work, we investigated the effects of aliovalent cation doping on the visible-light H2-evolution photocatalytic activity of the Ruddlesden–Popper layered perovskite oxynitride K2LaTa2O6N. The photocatalytic activity toward H2 evolution from an aqueous NaI solution was found to be enhanced by an increase in the specific surface area of the K2LaTa2O6N photocatalyst, which could be realized upon doping with lower-valence cations (e.g., Mg2+, Al3+, and Ga3+). Among the dopants examined at 1 mol % doping, Ga resulted in the highest activity. The activity of the Ga-doped specimen was further improved with increasing Ga concentration, where the maximal activity was obtained at 10 mol %, corresponding to an apparent quantum yield of 2.7 ± 0.4% at 420 nm from aqueous methanol. This number is the highest reported for a layered oxynitride photocatalyst. In the Ga-doped K2LaTa2O6N, a trade-off was observed between the Ga concentration and the photocatalytic activity. Although doping with Ga reduced the particle size of K2LaTa2O6N and suppressed undesirable charge recombination, it led to an enlarged bandgap, unsuitable for visible-light absorption. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 2041-1723 16 1 2025 Efficient and stable n-type sulfide overall water splitting with separated hydrogen production 8786 EN Haolin Luo Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University Zhixi Liu Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University Haifeng Lv State Key Laboratory of Precision and Intelligent Chemistry, School of Chemistry and Material Sciences, and Collaborative Innovation Center of Chemistry for Energy Materials (iChEM), University of Science and Technology of China Junie Jhon M. Vequizo Institute of Aqua Regeneration, Shinshu University Mengting Zheng College of Chemical and Biological Engineering, Zhejiang University Feng Han Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University Zhen Ye Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University Akira Yamakata Faculty of Natural Science and Technology, Okayama University Wenfeng Shangguan Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University Adam F. Lee Centre for Catalysis and Clean Energy, School of Environment and Science, Griffith University Xiaojun Wu State Key Laboratory of Precision and Intelligent Chemistry, School of Chemistry and Material Sciences, and Collaborative Innovation Center of Chemistry for Energy Materials (iChEM), University of Science and Technology of China Domen Kazunari Institute of Aqua Regeneration, Shinshu University Jun Lu College of Chemical and Biological Engineering, Zhejiang University Zhi Jiang Research Center for Combustion and Environment Technology, Shanghai Jiao Tong University N-type sulfide semiconductors are promising photocatalysts due to their broad visible-light absorption, facile synthesis and chemical diversity. However, photocorrosion and limited electron transport in one-step excitation and solid-state Z-scheme systems hinder efficient overall water splitting. Liquid-phase Z-schemes offer a viable alternative, but sluggish mediator kinetics and interfacial side reactions impede their construction. Here we report a stable Z-scheme system integrating n-type CdS and BiVO₄ with a [Fe(CN)₆]³⁻/[Fe(CN)₆]⁴⁻ mediator, achieving 10.2% apparent quantum yield at 450 nm with stoichiometric H₂/O₂ evolution. High activity reflects synergies between Pt@CrOx and Co3O4 cocatalysts on CdS, and cobalt-directed facet asymmetry in BiVO₄, resulting in matched kinetics for hydrogen and oxygen evolution in a reversible mediator solution. Stability is dramatically improved through coating CdS and BiVO4 with different oxides to inhibit Fe4[Fe(CN)6]3 precipitation and deactivation by a hitherto unrecognized mechanism. Separate hydrogen and oxygen production is also demonstrated in a two-compartment reactor under visible light and ambient conditions. This work unlocks the long-sought potential of n-type sulfides for efficient, durable and safe solar-driven hydrogen production. No potential conflict of interest relevant to this article was reported.

MDPI AG Acta Medica Okayama 2073-4344 14 10 2024 Photocatalytic Ammonia Decomposition Using Dye-Encapsulated Single-Walled Carbon Nanotubes 715 EN Tomoyuki Tajima Graduate School of Environmental and Life Science, Okayama University Kotone Yano Graduate School of Environmental and Life Science, Okayama University Kazushi Mukai Department of Materials Design and Engineering, University of Toyama Yutaka Takaguchi Department of Materials Design and Engineering, University of Toyama The photocatalytic decomposition of ammonia to produce N2 and H2 was achieved using single-walled carbon nanotube (SWCNT) nanohybrids. The physical modification of ferrocene-dye-encapsulated CNTs by amphiphilic C60-dendron yielded nanohybrids with a dye/CNT/C60 coaxial heterojunction. Upon irradiation with visible light, an aqueous solution of NH3 and dye@CNT/C60-dendron nanohybrids produced both N2 and H2 in a stoichiometric ratio of 1/3. The action spectra of this reaction clearly demonstrated that the encapsulated dye acted as the photosensitizer, exhibiting an apparent quantum yield (AQY) of 0.22% at 510 nm (the λmax of the dye). This study reports the first example of dye-sensitized ammonia decomposition and provides a new avenue for developing efficient and sustainable photocatalytic hydrogen production systems. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 1616-301X 2025 A Biologically-Architected Wear and Damage-Resistant Nanoparticle Coating From the Radular Teeth of Cryptochiton stelleri e21664 EN Taifeng Wang Department of Materials Science and Engineering, University of California Yu Chen Lyles School of Civil and Construction Engineering, Purdue University Ezra Sarmiento Department of Materials Science and Engineering, University of California Taige Hao Materials and Manufacturing Technologies Program, University of California Atsushi Arakaki Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology Michiko Nemoto Graduate School of Environmental and Life Science, Okayama University Pablo Zavattieri Lyles School of Civil and Construction Engineering, Purdue University David Kisailus Department of Materials Science and Engineering, University of California Nature utilizes simple building blocks to construct mechanically robust materials that demonstrate superior performance under extreme conditions. These exquisite structures result from the controlled synthesis and hierarchical assembly of nanoscale organic and mineral components that have provided critical evolutionary advantages to ensure survival. One such example is the ultrahard radular teeth found in mollusks, which are used to scrape against rock to feed on algae. Here, it is reported that the leading edges of these teeth consist of a wear-resistant coating that is comprised of densely packed ≈65 nm magnetic nanoparticles integrated within an organic matrix of chitin and protein. These mesocrystalline magnetite-based structures are assembled from smaller, highly aligned nanocrystals with inter/intracrystalline organics introduced during the crystallization process. Nanomechanical testing reveals that this multi-scale, nano-architected coating has a combination of increased hardness and a slight decrease in modulus versus geologic magnetite provides the surface of the chiton tooth with superior abrasion resistance. The mesocrystalline structures fracture at primary domain interfaces, corroborated by computational models, providing significant toughening to the tooth under extreme contact stresses. The design features revealed provide insight for the design and fabrication of next-generation advanced wear- and impact-resistant coatings for tooling, machinery, wind turbines, armor, etc. No potential conflict of interest relevant to this article was reported.

Royal Society of Chemistry (RSC) Acta Medica Okayama 1477-0520 23 27 2025 Fluorescence detection of DNA with a single-base mismatch by a Tm-independent peptide nucleic acid (PNA) twin probe 6557 6563 EN Koki Ishii Department of Applied Chemistry, Kindai University Hajime Shigeto Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) Shohei Yamamura Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST) Yoshitane Imai Department of Applied Chemistry, Kindai University Takashi Ohtsuki Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University Mizuki Kitamatsu Department of Applied Chemistry, Kindai University There is a need to develop efficient methods for detecting target nucleic acids to enable the rapid diagnosis and early treatment of diseases. We previously demonstrated that a peptide nucleic acid (PNA) twin probe, consisting of two PNAs each containing a fluorescent dye, with pyrene at one end, detects target DNA sequence-specifically through pyrene excimer emission. In this study, to advance the development of this probe system, we further investigated the fluorescence properties of the PNA twin probe P1 and P2, and found that the excimer fluorescence was significantly reduced when a mismatched base in the DNA sequence was present at the site of P1 closest to the pyrene. In other words, this probe was found to detect single-base mismatches without taking into account the thermal stability of the PNA/DNA hybrid. The detection limit of this PNA twin probe for the single-base-mismatched DNA was 2.7 nM. In the future, this probe should lead to a method to detect point mutations in endogenous nucleic acids within cells. No potential conflict of interest relevant to this article was reported.

Royal Society of Chemistry (RSC) Acta Medica Okayama 2398-4902 2025 Activation of barium titanate for photocatalytic overall water splitting via low-valence cation codoping EN Shigeru Ikeda Faculty of Science and Engineering, Konan University Kaori Takagi Faculty of Science and Engineering, Konan University Ryota Tomizawa Carbon Neutral Energy Development Division, Toyota Motor Corporation Tomoya Nagano Carbon Neutral Energy Development Division, Toyota Motor Corporation Koji Hayashi Carbon Neutral Energy Development Division, Toyota Motor Corporation Akira Yamakata Research Institute for Interdisciplinary Science, Okayama University Yoshitaro Nose Department of Materials Science and Engineering, Kyoto University Barium titanate (BaTiO3) has long been regarded as inactive for photocatalytic overall water splitting, in stark contrast to its perovskite counterparts SrTiO3 and CaTiO3. Here we report that BaTiO3 codoped with Al3+ and Sc3+ at Ti4+ sites under flux synthesis conditions is activated as a robust photocatalyst for overall water splitting. This material achieves apparent quantum yields of 29.8% at 310 nm and 27.5% at 365 nm, representing the first demonstration of efficient overall water splitting on BaTiO3. Comparative analyses show that BaTiO3 doped only with Al3+ suffers from severe band-edge disorder, whereas BaTiO3 codoped with Al3+ and Mg2+ exhibits clear activation with moderate efficiency. In contrast, BaTiO3 codoped with Al3+ and Sc3+ achieves the critical defect and structural control required to push the material across the threshold from inactive to highly active. These findings overturn the long-standing perception of BaTiO3 as unsuitable for water splitting and establish a general design principle for activating previously inactive perovskite oxides, thereby expanding the materials palette for solar-to-hydrogen energy conversion. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 5 2025 Current Status of Extracorporeal Membrane Oxygenation as a Treatment Strategy for Primary Graft Dysfunction after Lung Transplantation 329 337 EN Kei Matsubara Department of Thoracic Surgery, Hiroshima City Hiroshima Citizens Hospital Kentaroh Miyoshi Department of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shinichi Toyooka Department of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Review 10.18926/AMO/69433 Primary graft dysfunction (PGD) is one of the major risk factors affecting patients’ short- and long-term survival after lung transplantation. No particular management strategy has been established for PGD; supportive care is the mainstay of PGD treatment. When a supportive strategy fails, the patient may require the introduction of extracorporeal membrane oxygenation (ECMO) as the last-resort measure for severe PGD. A variety of study of ECMO as a PGD treatment was reported and the management of PGD patients developed so far. Early recognition of a patient’s need for ECMO and its prompt initiation are critical to improved outcomes. The use of venovenous-ECMO became the preferred procedure for PGD rather than venoarterial-ECMO. However, the current ECMO strategy has limitations, and using ECMO to manage patients with PGD is not sufficiently effective. Further studies are required to develop this promising technology. No potential conflict of interest relevant to this article was reported.

Ceramic Society of Japan Acta Medica Okayama 1348-6535 133 9 2025 Preparation and structural characterization of nanoporous silica/magnesium(II)-whitlockite composite particles 555 561 EN Takuya Kataoka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Daiki Hirota Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Eiji Fujii Industrial Technology Center of Okayama Prefecture Tomohiko Yoshioka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Satoshi Hayakawa Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University The preparation of particles composed of nanoporous silica (NS) and Mg2+-whitlockite (Mg-WH) would provide valuable insights for designing particles for biomedical applications. In this study, NS and Mg-WH composite particles were successfully synthesized. The addition of chitosan during synthesis possibly promoted the crystallization of calcium phosphate phases in the composite particles. Pore size distribution analysis of the particles showed a maximum at 3.2 nm. Investigating the adsorption of methylene blue onto the particles in a phosphate buffer (pH 7.4) showed that the saturated adsorption amount of methylene blue on the particles was significantly higher than that on commercial hydroxyapatite. The composite particles provided important results for potential applications as drug carriers for bone regeneration and repair. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 2590-1974 27 2025 Development of a technique to identify μm-sized organic matter in asteroidal material: An approach using machine learning 100277 EN Rahul Kumar The Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University Katsura Kobayashi The Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University Christian Potiszil The Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University Tak Kunihiro The Pheasant Memorial Laboratory, Institute for Planetary Materials, Okayama University Asteroidal materials contain organic matter (OM), which records a number of extraterrestrial environments and thus provides a record of Solar System processes. OM contain essential compounds for the origin of life. To understand the origin and evolution of OM, systematic identification and detailed observation using in-situ techniques is required. While both nm- and μm-sized OM were studied previously, only a small portion of a given sample surface was investigated in each study. Here, a novel workflow was developed and applied to identify and classify μm-sized OM on mm-sized asteroidal materials. The workflow involved image processing and machine learning, enabling a comprehensive and non-biased way of identifying, classifying, and measuring the properties of OM. We found that identifying OM is more accurate by classification with machine learning than by clustering. On the approach of classification with machine learning, five algorithms were tested. The random forest algorithm was selected as it scored the highest in 4 out of 5 accuracy parameters during evaluation. The workflow gave modal OM abundances that were consistent with those identified manually, demonstrating that the workflow can accurately identify 1-15 μm-sized OM. The size distribution of OM was modeled using the power-law distribution, giving slope α values that were consistent with fragmentation processes. The shape of the OM was quantified using circularity and solidity, giving a positive correlation and indicating these properties are closely related. Overall, the workflow enabled identification of many OM quickly and accurately and the obtainment of chemical and petrographic information. Such information can help the selection of OM for further in-situ techniques, and elucidate the origin and evolution of OM preserved in asteroidal materials. No potential conflict of interest relevant to this article was reported.

Oxford University Press (OUP) Acta Medica Okayama 1751-7362 18 1 2024 Cyanorhodopsin-II represents a yellow-absorbing proton-pumping rhodopsin clade within cyanobacteria wrae175 EN Masumi Hasegawa-Takano Atmosphere and Ocean Research Institute, The University of Tokyo Toshiaki Hosaka Laboratory for Protein Functional and Structural Biology, RIKEN Center for Biosystems Dynamics Research Keiichi Kojima Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yosuke Nishimura Atmosphere and Ocean Research Institute, The University of Tokyo Marie Kurihara Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yu Nakajima Atmosphere and Ocean Research Institute, The University of Tokyo Yoshiko Ishizuka-Katsura Laboratory for Protein Functional and Structural Biology, RIKEN Center for Biosystems Dynamics Research Tomomi Kimura-Someya Laboratory for Protein Functional and Structural Biology, RIKEN Center for Biosystems Dynamics Research Mikako Shirouzu Laboratory for Protein Functional and Structural Biology, RIKEN Center for Biosystems Dynamics Research Yuki Sudo Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Microbial rhodopsins are prevalent in many cyanobacterial groups as a light-energy-harvesting system in addition to the photosynthetic system. It has been suggested that this dual system allows efficient capture of sunlight energy using complementary ranges of absorption wavelengths. However, the diversity of cyanobacterial rhodopsins, particularly in accumulated metagenomic data, remains underexplored. Here, we used a metagenomic mining approach, which led to the identification of a novel rhodopsin clade unique to cyanobacteria, cyanorhodopsin-II (CyR-II). CyR-IIs function as light-driven outward H+ pumps. CyR-IIs, together with previously identified cyanorhodopsins (CyRs) and cyanobacterial halorhodopsins (CyHRs), constitute cyanobacterial ion-pumping rhodopsins (CyipRs), a phylogenetically distinct family of rhodopsins. The CyR-II clade is further divided into two subclades, YCyR-II and GCyR-II, based on their specific absorption wavelength. YCyR-II absorbed yellow light (λmax = 570 nm), whereas GCyR-II absorbed green light (λmax = 550 nm). X-ray crystallography and mutational analysis revealed that the difference in absorption wavelengths is attributable to slight changes in the side chain structure near the retinal chromophore. The evolutionary trajectory of cyanobacterial rhodopsins suggests that the function and light-absorbing range of these rhodopsins have been adapted to a wide range of habitats with variable light and environmental conditions. Collectively, these findings shed light on the importance of rhodopsins in the evolution and environmental adaptation of cyanobacteria. No potential conflict of interest relevant to this article was reported.

MDPI AG Acta Medica Okayama 1422-0067 26 10 2025 Stem Cell Factors BAM1 and WOX1 Suppressing Longitudinal Cell Division of Margin Cells Evoked by Low-Concentration Auxin in Young Cotyledon of Arabidopsis 4724 EN Yuli Jiang Institute for Translational Brain Reaearch, Fudan University Jian Liang Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences Chunyan Wang Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences Li Tan Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences Yoji Kawano Institute of Plant Science and Resources, Okayama University Shingo Nagawa Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences Highly differentiated tissues and organs play essential biological functions in multicellular organisms. Coordination of organ developmental process with tissue differentiation is necessary to achieve proper development of mature organs, but mechanisms for such coordination are not well understood. We used cotyledon margin cells from Arabidopsis plant as a new model system to investigate cell elongation and cell division during organ growth and found that margin cells endured a developmental phase transition from the “elongation” phase to the “elongation and division” phase at the early stage in germinating seedlings. We also discovered that the stem cell factors BARELY ANY MERISTEM 1 (BAM1) and WUSCHEL-related homeobox1 (WOX1) are involved in the regulation of margin cell developmental phase transition. Furthermore, exogenous auxin treatment (1 nanomolar,nM) promotes cell division, especially longitudinal cell division. This promotion of cell division did not occur in bam1 and wox1 mutants. Based on these findings, we hypothesized a new “moderate auxin concentration” model which emphasizes that a moderate auxin concentration is the key to triggering the developmental transition of meristematic cells. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1939-5981 2025 MATERIAL PROPERTIES OF DIE-CASTING DIE AROUND HEAT-CHECKING CREATED BY A HIGH-PRESSURE ALUMINUM ALLOY DIE-CASTING OPERATION EN Mitsuhiro Okayasu Department of Mechanical Systems Engineering, Okayama University Junya Shimazu Department of Mechanical Systems Engineering, Okayama University In this study, the material properties of a nitride die-casting die exhibiting heat-checking after the die-casting process were experimentally investigated using various methods. Based on the obtained results, the authors believe that several possible mechanisms underlying the formation of heat-checking can be identified. The microstructure of the die-casting die near the heat-checking region is characterized by equiaxed grains along the vicinity of the prior γ-grain boundaries, resulting from the lath martensitic formation. Additionally, numerous Cr–Mo–V-based nitride particles, approximately 100 nm in diameter, are precipitated. The surface hardness of the die-casting die, enhanced by nitriding, induces compressive residual stresses and increases adhesive forces. As a result of changes in microstructural characteristics and crack formation, the stress state near the die-casting die is altered, where compressive residual stresses, observed in the die-casting die, are released, leading to the tensile residual stresses. This phenomenon could accelerate the formation of a large number of heat-checking cracks. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0947-8396 131 9 2025 Optical and chemical properties of silver tree-like structure treated with gold galvanic substitution 744 EN Kazushi Honda Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Nobuyuki Takeyasu Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Galvanic gold substitution was executed in the presence of trisodium citrate on silver tree-like structures. No discernible difference in geometry was observed between the pre- and post-gold substitution phases, which benefited from the presence of citrate ions. The extent of gold substitution was regulated by the amount of gold ion solution added. After the gold substitution, an increase in extinction was observed in the ultraviolet region, indicating that gold was deposited at the surface. Raman scattering of para-toluenethiol was measured on the gold/silver tree-like structures at 488 nm excitations, where a decrease in the Raman peak intensity was observed as the quantity of gold ion solution increased. The results indicated that the optical property of silver was lost due to the increase of the amount of gold deposition. Concurrently, an investigation was conducted into the chemical resistance of the gold/silver tree-like structures, which was evaluated by measuring the resistivity inverse-proportional to the amount of silver ions dissolved by the diluted nitric acid. As the amount of gold ion solution added increased, the resistivity increased and became constant. The result implied that the surface chemical property had undergone a complete transformation into gold. No potential conflict of interest relevant to this article was reported.

Japan Society on Water Environment Acta Medica Okayama 1348-2165 23 2 2025 Study on the Removal Technology of Trichloramine from Drinking Water Using Ultraviolet Light 71 81 EN Ayumi Hashiguchi Faculty of Environmental, Life, Natural Science and Technology, Okayama University Shiho Yoshida Graduate School of Natural Science and Technology, Shimane University Shinya Echigo Graduate School of Global Environmental Studies, Kyoto University Ryohei Takanami Faculty of Design Technology, Osaka Sangyo University Hideaki Nagare Faculty of Environmental, Life, Natural Science and Technology, Okayama University Trichloramine (NCl3) is an inorganic chloramine that causes a pungent chlorine-like odor, and it is difficult to remove its precursors (nitrogen organic compounds and/or ammonia) completely from water. Powdered activated carbon, ozonation, and UV treatment have been applied for decomposing NCl3, but free chlorine was also decomposed. So, it is necessary to develop a technique that can selectively control NCl3 without losing free chlorine. UV light-emitting diodes (265, 280, and 300 nm) and plasma emission UV sheet (347 ± 52 nm, hereafter 350 nm) were compared to find the optimal wavelengths that decompose NCl3 but not free chlorine. As a result, 90.6, 96.7, 92.5, and 77.8% of NCl3 were removed at 265, 280, 300 (3,600 mJ/cm2), and 350 nm (14,400 mJ/cm2), respectively. On the other hand, free chlorine at neutral pH (hypochlorous acid is dominant) and slightly alkaline pH (hypochlorite ion is dominant) was not decomposed at 350 nm, but at other wavelengths (i.e., 265, 280, and 300 nm) the removals were more than 64%. Therefore, UV radiation at 350 nm can be candidates to remove NCl3 while maintaining free chlorine. However, this method requires high input energy, and further study is needed for evaluating the practical applicability of this method by considering optimal reactor design. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 1613-6810 2025 Polyglycerol‐Grafted Graphene Oxide with pH‐Responsive Charge‐Convertible Surface to Dynamically Control the Nanobiointeractions for Enhanced in Vivo Tumor Internalization 2503029 EN Yajuan Zou Research Institute for Interdisciplinary Science, Okayama University Alberto Bianco Research Institute for Interdisciplinary Science, Okayama University Yuta Nishina Research Institute for Interdisciplinary Science, Okayama University pH-responsive charge-convertible nanomaterials (NMs) ameliorate the treatment of cancer via simultaneously reducing nonspecific interactions during systemic circulation and improving targeted uptake within solid tumors. While promising, little is known about how the pH-responsiveness of charge-convertible NMs directs their interactions with biological systems, leading to compromised performance, including off-target retention and low specificity to tumor cells. In the present study, polyglycerol-grafted graphene oxide bearing amino groups (GOPGNH2) at different densities are reacted with dimethylmaleic anhydride (DMMA), a pH-responsive moiety, to generate a set of charge-convertible GOPGNH-DMMA variants. This permits the assessment of a quantitative correlation between the structure of GOPGNH-DMMA to their pH-responsiveness, their dynamic interactions with proteins and cells, as well as their in vivo biological fate. Through a systematic investigation, it is revealed that GOPGNH115-DMMA prepared from GOPGNH2 with higher amine density experienced fast charge conversion at pH 7.4 to induce non-specific interactions at early stages, whereas GOPGNH60-DMMA and GOPGNH30-DMMA prepared from lower amine density retarded off-target charge conversion to enhance tumor accumulation. Notably, GOPGNH60-DMMA is also associated with enough amounts of proteins under acidic conditions to promote in vivo tumor internalization. The findings will inform the design of pH-responsive NMs for enhanced treatment accuracy and efficacy. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1432-1068 35 1 2024 Association between lower limb muscle strength and musculoskeletal ambulation disability symptom complex in patients with medial meniscus posterior root tears 40 EN Mikao Fukuba Department of Physical Medicine and Rehabilitation, Okayama University Hospital Koki Kawada Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yuki Okazaki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yoshimi Katayama Department of Physical Medicine and Rehabilitation, Okayama University Hospital Masanori Hamada Department of Physical Medicine and Rehabilitation, Okayama University Hospital Toshifumi Ozaki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Takayuki Furumatsu Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Purpose In this study, we aimed to evaluate the changes in and the relationship between lower limb muscle strength and physical function before and after medial meniscus posterior root (MMPR) repair.<br> Methods Thirty-three patients who underwent MMPR repair were evaluated. Pain was evaluated with the numerical rating scale (NRS), and knee flexor/extensor muscle strength was assessed using a handheld dynamometer. Physical function was evaluated using a timed up and go (TUG) test. The NRS, knee flexor/extensor muscle strength, and TUG were compared preoperatively and 1 year postoperatively using the Wilcoxon signed-rank test. The correlation of patient characteristics, NRS score, knee flexor/extensor muscle strength, and preoperative TUG with the postoperative TUG was analyzed using Spearman’s correlation coefficient.<br> Results NRS (3.5 ± 2.1 to 0.1 ± 0.5 points), knee flexor strength (111.9 ± 50.2 to 146.7 ± 51.5 Nm), knee extensor strength (181.9 ± 92.8 to 256.9 ± 107.1 Nm), and TUG (12.3 ± 5.7 to 9.2 ± 2.2 s) all improved significantly from preoperatively to 1 year postoperatively (p < 0.001). The postoperative TUG was negatively correlated with the preoperative TUG (r = 0.578, p < 0.001), preoperative knee flexor muscle strength (r = − 0.355, p = 0.042), preoperative knee extensor muscle strength (r = − 0.437, p = 0.010), and postoperative knee extensor muscle strength (r = − 0.478, p = 0.004).<br> Conclusion In patients undergoing MMPR repair, surgery and rehabilitation significantly improve lower limb muscle strength and physical function. There was a significant correlation between lower limb muscle strength and TUG, and further strengthening of the lower limb muscles from the preoperative level is desirable to improve patients’ physical function further.<br> Level of evidence IV. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 4 2025 A Study of Periprosthetic Femoral Stem Fractures in Hip Arthroplasty for Femoral Neck Fracture 253 259 EN Yoshiaki Miyake Department of Orthopaedic Surgery, Japanese Red Cross Okayama Hospital Toru Takagi Department of Orthopaedic Surgery, Japanese Red Cross Okayama Hospital Taizo Konishiike Department of Orthopaedic Surgery, Japanese Red Cross Okayama Hospital Original Article 10.18926/AMO/69150 This study investigated the risk factors for bone fragility and perioperative periprosthetic femoral stem fractures in patients undergoing hip arthroplasty for femoral neck fractures. The records of 215 patients (42 male, 173 female; mean age, 84.4 years) were analyzed to assess correlations among periprosthetic fracture rates and sex, age, body mass index (BMI), Dorr classification, femoral stem fixation type (cemented/cementless), and bone mineral density (BMD) of the contralateral proximal femur. The overall prevalence of perioperative periprosthetic fractures was 4.7%. All patients with periprosthetic fractures were female, and all but one were ≥ 80 years of age. Fracture rates were higher in patients with lower BMI, although this difference was not significant. The fracture rates were 0%, 4.7%, and 7.9% for Dorr types A, B, and C, respectively, and 0% and 5.3% for patients who received cemented and cementless stems, respectively. The findings indicated that female patients, those of advanced age, those with lower BMI, and those with Dorr type C had lower BMDs. Although BMD was significantly lower in patients who received cemented stems compared to those who received cementless stems, no fractures were observed in the former group, suggesting that the use of cemented stems is safe for this high-risk population. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 4 2025 Bloodstream Infections Caused by Gram-Negative Bacteria in Geriatric Patients: Epidemiology, Antimicrobial Resistance and The Factors Affecting Mortality 231 242 EN M Enes Kardan Department of Infectious Diseases, Faculty of Medicine, Namik Kemal University Ilknur Erdem Department of Infectious Diseases, Faculty of Medicine, Namik Kemal University Emre Yildiz Department of Infectious Diseases, Faculty of Medicine, Namik Kemal University Nuri Kiraz Department of Medical Microbiology, Faculty of Medicine, Namik Kemal University Aliye Çelikkol Department of Biochemistry, Faculty of Medicine, Namik Kemal University Original Article 10.18926/AMO/69148 Bloodstream infections (BSIs) are an important cause of morbidity and mortality in geriatric patients. We retrospectively analyzed the cases of geriatric patients who developed BSIs due to gram-negative bacteria in order to evaluate the epidemiology, antimicrobial resistance, and the factors affecting mortality. The cases of 110 patients aged ≥ 65 years admitted to our hospital between January 1, 2017, and December 31, 2022 were assessed; 70 (63.6%) of the BSIs were healthcare-associated BSIs. The urinary system was the most common detectable source of infection at 43.6%. The most frequently isolated bacteria were Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae, in that order. Carbapenem resistance was detected in 17 patients (15.5%), and extended-spectrum beta-lactamase (ESBL) production from Enterobacterales family members was detected in 37 (51.4%) patients. Multivariate analysis revealed that (i) the probability of mortality in the patients with total bilirubin was increased by approx. sixfold and (ii) the likelihood of mortality for those with a Pitt bacteremia score (PBS) ≥ 4 points was approx. 17 times higher. PBS and simplified qPitt scores can help predict mortality and manage geriatric patients. There is a significant increase in mortality among patients with procalcitonin (PCT) levels at ≥ 2 nm/ml. No potential conflict of interest relevant to this article was reported.

MDPI AG Acta Medica Okayama 1999-4923 17 7 2025 Development of an Antimicrobial Coating Film for Denture Lining Materials 902 EN Kumiko Yoshihara National Institute of Advanced Industrial Science and Technology (AIST), Health and Medical Research Institute Takeru Kameyama Graduate School of Natural Science and Technology, Okayama University Noriyuki Nagaoka Dental School, Advanced Research Center for Oral and Craniofacial Science, Okayama University Yukinori Maruo Department of Prosthodontics, Okayama University Yasuhiro Yoshida Department of Biomaterials and Bioengineering, Faculty of Dental Medicine, Hokkaido University Bart Van Meerbeek BIOMAT, Department of Oral Health Sciences, KU Leuvem Takumi Okihara Graduate School of Natural Science and Technology, Okayama University Background/Objectives: Denture hygiene is essential for the prevention of oral candidiasis, a condition frequently associated with Candida albicans colonization on denture surfaces. Cetylpyridinium chloride (CPC)-loaded montmorillonite (CPC-Mont) has demonstrated antimicrobial efficacy in tissue conditioners and demonstrates potential for use in antimicrobial coatings. In this study, we aimed to develop and characterize CPC-Mont-containing coating films for dentures, focusing on their physicochemical behaviors and antifungal efficacies. Methods: CPC was intercalated into sodium-type montmorillonite to prepare CPC-Mont; thereafter, films containing CPC-Mont were fabricated using emulsions of different polymer types (nonionic, cationic, and anionic). CPC loading, release, and recharging behaviors were assessed at various temperatures, and activation energies were calculated using Arrhenius plots. Antimicrobial efficacy against Candida albicans was evaluated for each film using standard microbial assays. Results: X-ray diffraction analysis confirmed the expansion of montmorillonite interlayer spacing by approximately 3 nm upon CPC loading. CPC-Mont showed temperature-dependent release and recharging behavior, with higher temperatures enhancing its performance. The activation energy for CPC release was 38 kJ/mol, while that for recharging was 26 kJ/mol. Nonionic emulsions supported uniform CPC-Mont dispersion and successful film formation, while cationic and anionic emulsions did not. CPC-Mont-containing coatings maintained antimicrobial activity against Candida albicans on dentures. Conclusions: CPC-Mont can be effectively incorporated into nonionic emulsion-based films to create antimicrobial coatings for denture applications. The films exhibited temperature-responsive, reversible CPC release and recharging behaviors, while maintaining antifungal efficacy, findings which suggest the potential utility of CPC-Mont-containing films as a practical strategy to prevent denture-related candidiasis. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0021-9258 301 7 2025 A repertoire of visible light–sensitive opsins in the deep-sea hydrothermal vent shrimp Rimicaris hybisae 110291 EN Yuya Nagata Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Norio Miyamoto Institute for Extra-Cutting-Edge Science and Technology Avant-Garde Research (X-Star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Keita Sato Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yosuke Nishimura Research Center for Bioscience and Nanoscience (CeBN), Research Institute for Marine Resources Utilization, Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Yuki Tanioka School of Pharmaceutical Sciences, Okayama University Yuji Yamanaka School of Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Kuto Takahashi Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Kohei Obayashi Department of Biology, Graduate School of Science, Kobe University Hisao Tsukamoto Department of Biology, Graduate School of Science, Kobe University Ken Takai Institute for Extra-Cutting-Edge Science and Technology Avant-Garde Research (X-Star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Hideyo Ohuchi Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takahiro Yamashita Department of Biophysics, Graduate School of Science, Kyoto University Yuki Sudo Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Keiichi Kojima Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Unlike terrestrial environments, where humans reside, there is no sunlight in the deep sea. Instead, dim visible light from black-body radiation and bioluminescence illuminates hydrothermal vent areas in the deep sea. A deep-sea hydrothermal vent shrimp, Rimicaris hybisae, is thought to detect this dim light using its enlarged dorsal eye; however, the molecular basis of its photoreception remains unexplored. Here, we characterized the molecular properties of opsins, universal photoreceptive proteins in animals, found in R. hybisae. Transcriptomic analysis identified six opsins: three Gq-coupled opsins, one Opn3, one Opn5, and one peropsin. Functional analysis revealed that five of these opsins exhibited light-dependent G protein activity, whereas peropsin exhibited the ability to convert all-trans-retinal to 11-cis-retinal like photoisomerases. Notably, all the R. hybisae opsins, including Opn5, convergently show visible light sensitivity (around 457–517 nm), whereas most opsins categorized as Opn5 have been demonstrated to be UV sensitive. Mutational analysis revealed that the unique visible light sensitivity of R. hybisae Opn5 is achieved through the stabilization of a protonated Schiff base by a counterion residue at position 83 (Asp83), which differs from the position identified in other opsins. These findings suggest that the vent shrimp R. hybisae has adapted its photoreceptive devices to dim deep-sea hydrothermal light by selectively maintaining a repertoire of visible light–sensitive opsins, including the uniquely tuned Opn5. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 2 2025 Effectiveness of Postoperative Irradiation in Patients with cN0 Early Breast Cancer Treated with Sentinel Lymph Node Surgery 101 107 EN Hiroshi Isozaki Department of Surgery, Oomoto Hospital Sasau Matsumoto Department of Surgery, Oomoto Hospital Takehiro Takama Department of Surgery, Oomoto Hospital Yuka Isozaki Department of Surgery, Oomoto Hospital Original Article 10.18926/AMO/68648 To evaluate the effectiveness of postoperative irradiation (POI) for patients with cN0 early breast cancer, we retrospectively analyzed the cases of 650 consecutive breast cancer patients who underwent sentinel lymph node (SLN)-guided surgery (2005-2022) at our hospital. In this cohort, 53% (278/521) of the patients who underwent breast conservative surgery (BCS) and 96% (124/129) of those treated with mastectomy did not receive POI. The patients who underwent BCS were treated with POI using opposing tangential field irradiation. A false negative (FN) SLN was retrospectively defined as a negative metastasis in SLN plus positive recurrence in the axillary lymph nodes. Recurrence was detected in 83 patients. A logistic regression analysis revealed that the nuclear grade (odds ratio [OR] 1.69), POI (OR 0.41), and postoperative hormone therapy (OR 0.40) were each significantly related to recurrence. The 26.1% (12/46) FN rate of the non-POI patients decreased to 5.8% (1/17) compared to those treated with POI. The rate of axillary recurrence was significantly lower in the POI group (0.4%) versus the non-POI group (2.7%) (p=0.0355). The rate of locoregional recurrence was also significantly lower in the POI group (2.0%) versus the non-POI group (13.4%) (p<0.0001). No significant difference was observed in the rate of distant recurrence between the POI (4.0%) and non-POI (3.3%) (p=0.831) groups. These results indicated that the postoperative opposing tangential field irradiation of conserved breast tissue inhibited recurrence in the axillary lymph nodes. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 2 2025 Association between the Pretreatment Body Mass Index and Anamorelin’s Efficacy in Patients with Cancer Cachexia: A Retrospective Cohort Study 65 73 EN Masatoshi Maki Department of Hospital Pharmacy, NHO Fukuyama Medical Center Ryo Takada Department of Hospital Pharmacy, NHO Fukuyama Medical Center Tomoyuki Ishigo Department of Pharmacy, Sapporo Medical University Hospital Miki Fujiwara Department of Hospital Pharmacy, NHO Fukuyama Medical Center Yoko Takahashi Department of Hospital Pharmacy, NHO Fukuyama Medical Center Shinya Otsuka Department of Surgery, NHO Fukuyama Medical Center Koji Tamura Department of Hospital Pharmacy, NHO Fukuyama Medical Center Terutaka Hamaoka Department of Hospital Pharmacy, NHO Fukuyama Medical Center Original Article 10.18926/AMO/68644 Anamorelin (ANAM) is used to treat cancer-associated cachexia, a syndrome involving muscle loss and anorexia. The timing of the initiation of ANAM treatment is crucial to its efficacy. Although the body mass index (BMI) is a diagnostic criterion for cancer cachexia, no studies have explored its association with ANAM efficacy. We conducted a single-center, retrospective cohort study to investigate the association between the pre-treatment BMI and ANAM efficacy in patients with cancer-associated cachexia (n=47). The ANAM treatment was considered effective if the patient’s appetite improved within 30 days of treatment initiation. We calculated a BMI cutoff value (19.5 kg/m2) and used it to divide the patients into high- and low-BMI groups. Their background, clinical laboratory values, cancer types, and treatment lines were investigated. Twenty (42.6%) had a high BMI (≥ 19.5 kg/m2) and 27 (57.4%) had a low BMI (< 19.5 kg/m2). High BMI was significantly associated with ANAM effectiveness (odds ratio 7.86, 95% confidence interval 1.99-31.00, p=0.003). Together these results indicate that it is beneficial to initiate ANAM treatment before a patient’s BMI drops below 19.5 kg/m2. Our findings will help advance cancer cachexia treatment and serve as a reference for clinicians to predict ANAM’s efficacy. No potential conflict of interest relevant to this article was reported.

Beilstein-Institut Acta Medica Okayama 1860-5397 21 2025 Photochemically assisted synthesis of phenacenes fluorinated at the terminal benzene rings and their electronic spectra 670 679 EN Yuuki Ishii Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Minoru Yamaji Division of Molecular Science, Graduate School of Science and Engineering, Gunma University Fumito Tani Institute for Materials Chemistry and Engineering, Kyushu University Kenta Goto Institute for Materials Chemistry and Engineering, Kyushu University Yoshihiro Kubozono Research Institute for Interdisciplinary Science, Okayama University Hideki Okamoto Graduate School of Environmental, Life, Natural Science and Technology, Okayama University [n]Phenacenes ([n] = 5-7), octafluorinated at the terminal benzene rings (F8-phenacenes: F8PIC, F8FUL, and F87PHEN), were photochemically synthesized, and their electronic spectra were investigated to reveal the effects of the fluorination on the electronic features of phenacene molecules. F8-Phenacenes were conveniently synthesized by the Mallory photoreaction of the corresponding fluorinated diarylethenes as the key step. Upon fluorination on the phenacene cores, the absorption and fluorescence bands of the F8-phenacenes in CHCl3 systematically red-shifted by ca. 3-5 nm compared to those of the corresponding parent phenacenes. The vibrational progressions of the absorption and fluorescence bands were little affected by the fluorination in the solution phase. In the solid state, the absorption band of F8-phenacenes appeared in the similar wavelength region for the corresponding parent phenacenes whereas their fluorescence bands markedly red-shifted and broadened. These observations suggest that the intermolecular interactions of excited-state F8-phenacene molecules are significantly different from those of the corresponding parent molecules, most likely due to different crystalline packing motifs. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2076-3417 14 23 2024 Preparation of Nano- and Microparticles Obtained from Polymerization Reaction and Their Application to Surface Coating of Woody Materials 11326 EN Toshinori Shimanouchi Department of Environmental Chemistry and Materials, Okayama University, Daichi Hirota Department of Environmental Chemistry and Materials, Okayama University, Masafumi Yoshida Department of Environmental Chemistry and Materials, Okayama University, Kazuma Yasuhara Division of Materials Science, Nara Institute of Science and Technology (NAIST) Yukitaka Kimura Department of Environmental Chemistry and Materials, Okayama University, A surface coating of polymer particles of different hydrophobicity and wide-ranged size is helpful for the surface modification of materials such as woody thin board (WTB) derived from biomass. A preparation method for polymer particles was, in this study, proposed using a capillary-type flow system. Under hydrothermal conditions, the refinement of dispersed oil droplets in water (O/W emulsions) and the polymerization reaction could be simultaneously advanced, and polymer particles of polystyrene (PS), polyvinyl alcohol (PVA), polymethyl methacrylate (PMMA), and poly-L-lactic acid (PLLA) with a particle size of about 100 nm could be synthesized. The coating of polymer particles gave an improved effect on the water repellency of WTBs due to the hydrophobicity of polymer particles and an alteration of surface roughness, and it also provided long-term stability (more than 6 years). No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1661-6596 25 22 2024 Distribution and Incorporation of Extracellular Vesicles into Chondrocytes and Synoviocytes 11942 EN Takashi Ohtsuki Department of Medical Technology, Graduate School of Health Sciences, Okayama University Ikumi Sato Department of Medical Technology, Graduate School of Health Sciences, Okayama University Ren Takashita Department of Medical Technology, Graduate School of Health Sciences, Okayama University Shintaro Kodama Department of Medical Technology, Graduate School of Health Sciences, Okayama University Kentaro Ikemura Department of Medical Technology, Graduate School of Health Sciences, Okayama University Gabriel Opoku Department of Medical Technology, Graduate School of Health Sciences, Okayama University Shogo Watanabe Department of Medical Technology, Graduate School of Health Sciences, Okayama University Takayuki Furumatsu Department of Orthopedic Surgery, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Hiroshi Yamada Department of Neuroscience, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Mitsuru Ando Laboratory of Biomaterials, Institute for Life and Medical Sciences, Kyoto University Kazunari Akiyoshi Department of Immunology, Graduate School of Medicine, Kyoto University Keiichiro Nishida Department of Orthopedic Surgery, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Satoshi Hirohata Department of Medical Technology, Graduate School of Health Sciences, Okayama University Osteoarthritis (OA) is a chronic disease affecting over 500 million people worldwide. As the population ages and obesity rates rise, the societal burden of OA is increasing. Pro-inflammatory cytokines, particularly interleukin-1β, are implicated in the pathogenesis of OA. Recent studies suggest that crosstalk between cartilage and synovium contributes to OA development, but the mechanisms remain unclear. Extracellular vesicles (EVs) were purified from cell culture-conditioned medium via ultracentrifugation and confirmed using transmission electron microscopy, nanoparticle tracking analysis, and western blotting. We demonstrated that EVs were taken up by human synoviocytes and chondrocytes in vitro, while in vivo experiments revealed that fluorescent-labelled EVs injected into mouse joints were incorporated into chondrocytes and synoviocytes. EV uptake was significantly inhibited by dynamin-mediated endocytosis inhibitors, indicating that endocytosis plays a major role in this process. Additionally, co-culture experiments with HEK-293 cells expressing red fluorescent protein (RFP)-tagged CD9 and the chondrocytic cell line OUMS-27 confirmed the transfer of RFP-positive EVs across a 600-nm but not a 30-nm filter. These findings suggest that EVs from chondrocytes are released into joint fluid and taken up by cells within the cartilage, potentially facilitating communication between cartilage and synovium. The results underscore the importance of EVs in OA pathophysiology. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 0947-6539 30 70 2024 MoSe2-Sensitized Water Splitting Assisted by C60-Dendrons on the Basal Surface e202402690 EN Tomoyuki Tajima Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Tomoki Matsuura Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Arif Efendi Department of Materials Design and Engineering, University of Toyama Mariko Yukimoto Department of Materials Design and Engineering, University of Toyama Yutaka Takaguchi Department of Materials Design and Engineering, University of Toyama To facilitate water splitting using MoSe2 as a light absorber, we fabricated water-dispersible MoSe2/C60-dendron nanohybrids via physical modification of the basal plane of MoSe2. Upon photoirradiation, the mixed-dimension MoSe2/C60 (2D/0D) heterojunction generates a charge-separated state (MoSe2⋅+/C60⋅−) through electron extraction from the exciton in MoSe2 to C60. This process is followed by the hydrogen evolution reaction (HER) from water in the presence of a sacrificial donor (1-benzyl-1,4-dihydronicotinamide) and co-catalyst (Pt-PVP). The apparent quantum yields of the HER were estimated to be 0.06 % and 0.27 % upon photoexcitation at the A- and B-exciton absorption peaks (λmax=800 and 700 nm), respectively. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2079-4991 14 20 2024 Colossal Dielectric Constant of Nanocrystalline/Amorphous Homo-Composite BaTiO3 Films Deposited via Pulsed Laser Deposition Technique 1677 EN Shinya Kondo Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Taichi Murakami Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Loick Pichon Institut National de la Recherche Scientifique (INRS), Centre Énergie, Matériaux et Télécommunications Joel Leblanc-Lavoie Institut National de la Recherche Scientifique (INRS), Centre Énergie, Matériaux et Télécommunications Takashi Teranishi Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Akira Kishimoto Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Khakani, My Ali El Khakani Institut National de la Recherche Scientifique (INRS), Centre Énergie, Matériaux et Télécommunications We report the pulsed laser deposition (PLD) of nanocrystalline/amorphous homo-composite BaTiO3 (BTO) films exhibiting an unprecedented combination of a colossal dielectric constant (epsilon(r)) and extremely low dielectric loss (tan delta). By varying the substrate deposition temperature (T-d) over a wide range (300-800 degrees C), we identified T-d = 550 degrees C as the optimal temperature for growing BTO films with an epsilon(r) as high as similar to 3060 and a tan delta as low as 0.04 (at 20 kHz). High-resolution transmission electron microscopy revealed that the PLD-BTO films consist of BTO nanocrystals (similar to 20-30 nm size) embedded within an otherwise amorphous BTO matrix. The impressive dielectric behavior is attributed to the combination of highly crystallized small BTO nanograins, which amplify interfacial polarization, and the surrounding amorphous matrix, which effectively isolates the nanograins from charge carrier transport. Our findings could facilitate the development of next-generation integrated dielectric devices. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 78 5 2024 Effect of Radon Inhalation on Murine Brain Proteins: Investigation Using Proteomic and Multivariate Analyses 387 399 EN Shota Naoe Graduate School of Health Sciences, Okayama University Ayumi Tanaka Graduate School of Health Sciences, Okayama University Norie Kanzaki Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency Reiju Takenaka Graduate School of Health Sciences, Okayama University Akihiro Sakoda Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency Takaaki Miyaji Advanced Science Research Center, Okayama University Kiyonori Yamaoka Faculty of Health Sciences, Okayama University Takahiro Kataoka Faculty of Health Sciences, Okayama University Original Article 10.18926/AMO/67663 Radon is a known risk factor for lung cancer; however, it can be used beneficially, such as in radon therapy. We have previously reported the enhancement of antioxidant effects associated with trace amounts of oxidative stress as one of the positive biological effects of radon inhalation. However, the biological effects of radon inhalation are incompletely understood, and more detailed and comprehensive studies are required. Although several studies have used proteomics to investigate the effects of radon inhalation on body proteins, none has focused on brain proteins. In this study, we evaluated the expression status of proteins in murine brains using proteomic and multivariate analyses to identify those whose expressions changed following two days of radon inhalation at a concentration of 1,500 Bq/m3. We found associations of radon inhalation with the expressions of seven proteins related to neurotransmission and heat shock. These proteins may be proposed as biomarkers indicative of radon inhalation. Although further studies are required to obtain the detailed biological significance of these protein alterations, this study contributes to the elucidation of the biological effects of radon inhalation as a low-dose radiation. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 78 4 2024 Multicenter Remote-Access Simulation of Vaginal Delivery for High-Flexibility Medical Education during the Coronavirus Pandemic 313 322 EN Eriko Eto Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Jota Maki Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Noriyuki Yamashita Center for Education in Medicine and Health Sciences, Okayama University Toru Hasegawa Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ayano Suemori Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hikari Nakato Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hikaru Oba Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomohiro Mitoma Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Sakurako Mishima Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Satoe Kirino Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Akiko Ohira Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hisashi Masuyama Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/67547 During the coronavirus pandemic, face-to-face simulation education became impossible. Therefore, we aimed to develop remote-access simulation education with a sense of realism through Information and Communication Technology (ICT) using a perinatal whole-body management and delivery simulator. In September 2021, we administered a multi-center simultaneous remote simulation based on our developed model. Ten universities in the Chugoku–Shikoku region were connected via a web-conferencing system to a live broadcast of a virtual vaginal birth in which a fictional hospitalized pregnant woman experienced accelerated labor and gave birth through vacuum delivery for fetal distress. A Video on Demand (VOD) was made beforehand using a new simulator that allowed for a visual understanding of the process of the inter-vaginal examination. We provided a participatory program that enhanced the sense of realism by combining VOD and real-time lectures on each scenario, with two-way communication between participants and trainee doctors using a chat function. Most participants answered “satisfied” or “very satisfied” with the content, level of difficulty, and level of understanding. From November 2021, we have used the videos of all processes in face-to-face classes. Our construction of a high-flexibility education system using remote simulation in the field of obstetrics and gynecology, especially in the vaginal delivery module, is unique, creative, and sustainable. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 78 4 2024 Can Pelvic Lymph Node Dissection in Prostate Cancer Patients with a 5% Briganti Nomogram Cut-off Value Provide an Oncological Benefit? A Large Multi-Institutional Cohort Study in Japan 307 312 EN Naoya Sugihara Department of Urology, Ehime University Katsuyoshi Hashine Department of Urology, National Hospital Organization Shikoku Cancer Center Natsumi Yamashita Division of Epidemiology, National Hospital Organization Shikoku Cancer Center Miki Sakamoto Department of Urology, Ehime University Masato Terashita Department of Urology, Ehime University Keisuke Funaki Department of Urology, Ehime University Kaori Saiki Department of Urology, Ehime University Takatora Sawada Department of Urology, Ehime University Toshio Kakuda Department of Urology, Ehime University Kenichi Nishimura Department of Urology, Ehime University Tetsuya Fukumoto Department of Urology, Ehime University Noriyosi Miura Department of Urology, Ehime University Yuki Miyauchi Department of Urology, Ehime University Tadahiko Kikugawa Department of Urology, Ehime University Takashi Saika Department of Urology, Ehime University Original Article 10.18926/AMO/67546 The Briganti nomogram (cut-off value 5%) is commonly used to determine the indications for pelvic lymph node dissection (PLND) in patients with prostate cancer. We retrospectively analyzed the potential oncological benefit of PLND based on the 5% cut-off value on the Briganti nomogram. We obtained the data from the Medical Investigation Cancer Network (MICAN) Study, which included 3,463 patients who underwent a radical prostatectomy (RP) at nine institutions in Japan between 2010 and 2020. We included patients with Briganti scores ≥ 5% and a follow-up period ≥6 months and excluded patients categorized in the very high-risk group (based on NCCN categories); a final total of the cases of 1,068 patients were analyzed. The biochemical recurrence (BCR)-free survival was significantly worse in the patients who underwent PLND compared to those who did not (p=0.019). A multivariate analysis showed that high prostate-specific antigen (PSA) levels (p<0.001) and an advanced T-stage (p=0.018) were significant prognostic factors for BCR, whereas PLND had no effect on BCR (p=0.059). Thus, PLND in patients with prostate cancer whose Briganti score was 5% did not provide any oncological benefit. Further research is necessary to determine the indication criteria for conducting PLND. No potential conflict of interest relevant to this article was reported.

Nature Portfolio Acta Medica Okayama 2041-1723 15 1 2024 Controlling 229Th isomeric state population in a VUV transparent crystal 5536 EN Takahiro Hiraki Research Institute for Interdisciplinary Science, Okayama University Koichi Okai Research Institute for Interdisciplinary Science, Okayama University Michael Bartokos Institute for Atomic and Subatomic Physics, TU Wien Kjeld Beeks Institute for Atomic and Subatomic Physics, TU Wien Hiroyuki Fujimoto National Institute of Advanced Industrial Science and Technology (AIST) Yuta Fukunaga Research Institute for Interdisciplinary Science, Okayama University Hiromitsu Haba RIKEN Yoshitaka Kasamatsu Graduate School of Science, Osaka University Shinji Kitao Institute for Integrated Radiation and Nuclear Science, Kyoto University Adrian Leitner Institute for Atomic and Subatomic Physics, TU Wien Takahiko Masuda Research Institute for Interdisciplinary Science, Okayama University Ming Guan Research Institute for Interdisciplinary Science, Okayama University Nobumoto Nagasawa Japan Synchrotron Radiation Research Institute Ryoichiro Ogake Research Institute for Interdisciplinary Science, Okayama University Martin Pimon Institute for Atomic and Subatomic Physics, TU Wien Martin Pressler Institute for Atomic and Subatomic Physics, TU Wien Noboru Sasao Research Institute for Interdisciplinary Science, Okayama University Fabian Schaden Institute for Atomic and Subatomic Physics, TU Wien Thorsten Schumm Institute for Atomic and Subatomic Physics, TU Wien Makoto Seto Institute for Integrated Radiation and Nuclear Science, Kyoto University Yudai Shigekawa RIKEN Kotaro Shimizu Research Institute for Interdisciplinary Science, Okayama University Tomas Sikorsky Institute for Atomic and Subatomic Physics, TU Wien Kenji Tamasaku RIKEN SPring-8 Center Sayuri Takatori Research Institute for Interdisciplinary Science, Okayama University Tsukasa Watanabe National Institute of Advanced Industrial Science and Technology (AIST) Atsushi Yamaguchi RIKEN Yoshitaka Yoda Japan Synchrotron Radiation Research Institute Akihiro Yoshimi Research Institute for Interdisciplinary Science, Okayama University Koji Yoshimura Research Institute for Interdisciplinary Science, Okayama University The radioisotope thorium-229 (Th-229) is renowned for its extraordinarily low-energy, long-lived nuclear first-excited state. This isomeric state can be excited by vacuum ultraviolet (VUV) lasers and Th-229 has been proposed as a reference transition for ultra-precise nuclear clocks. To assess the feasibility and performance of the nuclear clock concept, time-controlled excitation and depopulation of the Th-229 isomer are imperative. Here we report the population of the Th-229 isomeric state through resonant X-ray pumping and detection of the radiative decay in a VUV transparent Th-229-doped CaF2 crystal. The decay half-life is measured to 447(25) s, with a transition wavelength of 148.18(42) nm and a radiative decay fraction consistent with unity. Furthermore, we report a new "X-ray quenching" effect which allows to de-populate the isomer on demand and effectively reduce the half-life. Such controlled quenching can be used to significantly speed up the interrogation cycle in future nuclear clock schemes. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1474-905X 23 8 2024 Intramolecular [π4s + π4s] photocycloaddition of carbon- and nitrogen-bridged [32](1,4)naphthalenophanes 1509 1519 EN Yukiko Oguma Division of Earth, Life, and Molecular Sciences, Graduate School of Natural Science and Technology, Okayama University Masanori Yamamoto Division of Earth, Life, and Molecular Sciences, Graduate School of Natural Science and Technology, Okayama University Yukinari Sunatsuki Division of Earth, Life, and Molecular Sciences, Graduate School of Natural Science and Technology, Okayama University Hiromi Ota Department of Instrumental Analysis, Advanced Science Research Center, Okayama University Minoru Yamaji Division of Molecular Science, Graduate School of Science and Engineering, Gunma University Hideki Okamoto Division of Earth, Life, and Molecular Sciences, Graduate School of Natural Science and Technology, Okayama University [32](1,4)Naphthalenophanes, bearing carbon-bridge chains (syn- and anti-NPs) and nitrogen-bridge chains (syn- and anti-ANPs), were synthesized, and their X-ray structures and photoreactions were investigated. The intramolecular separation distance between the naphthalene cores for ANPs was shorter than that for NPs, suggesting that intramolecular interactions between the naphthalene rings were more efficient for ANPs compared to NPs. Upon photoirradiation at 300 nm, anti-NP, syn-ANP and anti-ANP produced the corresponding intramolecular [π4s + π4s] cycloadducts, whereas syn-NP gave an unidentified complex product mixture. Quantum yields for the photo-consumption (ΦPC) of NPs and ANPs were evaluated to quantitatively compare their photoreactivity. The ΦPC values of ANPs were approximately two-fold higher than those of ANPs.Noteworthily, the ΦPC value of syn-ANP was estimated to be unity. Based on these results we discuss the effects of the alignments of the naphthalene cores (anti vs. syn) and the bridging elements (C-bridge vs. N-bridge) on the photoreaction efficiencies of [32](1,4)naphthalenophanes. No potential conflict of interest relevant to this article was reported.

American Chemical Society (ACS) Acta Medica Okayama 1520-6106 128 27 2024 Bidirectional Optical Control of Proton Motive Force in Escherichia coli Using Microbial Rhodopsins 6509 6517 EN Kotaro Nakanishi Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Keiichi Kojima Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yoshiyuki Sowa Department of Frontier Bioscience and Research Center for Micro-Nano Technology, Hosei University Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Proton (H+) motive force (PMF) serves as the energy source for the flagellar motor rotation, crucial for microbial motility. Here, to control PMF using light, we introduced light-driven inward and outward proton pump rhodopsins, RmXeR and AR3, into Escherichia coli. The motility of E. coli cells expressing RmXeR and AR3 significantly decreased and increased upon illumination, respectively. Tethered cell experiments revealed that, upon illumination, the torque of the flagellar motor decreased to nearly zero (28 pN nm) with RmXeR, while it increased to 1170 pN nm with AR3. These alterations in PMF correspond to +146 mV (RmXeR) and −140 mV (AR3), respectively. Thus, bidirectional optical control of PMF in E. coli was successfully achieved by using proton pump rhodopsins. This system holds a potential for enhancing our understanding of the roles of PMF in various biological functions. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 78 3 2024 The Role of the Lipid Profile and Oxidative Stress in Fatigue, Sleep Disorders and Cognitive Impairment in Patients with Multiple Sclerosis 259 270 EN Gonul Vural Department of Neurology, Faculty of Medicine, Ankara Yildirim Beyazit University Esra Demir Department of Neurology, Ankara City Hospital Sadiye Gumusyayla Department of Neurology, Faculty of Medicine, Ankara Yildirim Beyazit University Funda Eren Department of Clinical Biochemistry, Ankara City Hospital Serdar Barakli Department of Neurology, Ankara City Hospital Salim Neselioglu Department of Clinical Biochemistry, Ankara City Hospital Ozcan Erel Department of Clinical Biochemistry, Ankara City Hospital Original Article 10.18926/AMO/67201 The aim of this study is to investigate the relationship of the lipid profile, dysfunctional high-density lipoprotein, ischaemia-modified albumin and thiol–disulfide homeostasis with cognitive impairment, fatigue and sleep disorders in patients with multiple sclerosis. The cognitive functions of patients were evaluated with the Brief International Cognitive Assessment for Multiple Sclerosis battery. Fatigue was evaluated with the Fatigue Severity Scale and the Fatigue Impact Scale. The Pittsburgh Sleep Quality Index and the Epworth Sleepiness Scale were used to assess patients’ sleep disturbance. Peripheral blood samples were collected, and lipid levels and myeloperoxidase and paraoxonase activity were measured. The myeloperoxidase/paraoxonase ratio, which indicates dysfunctional high-density lipoprotein, was calculated. Thiol–disulfide homeostasis and ischaemia-modified albumin were measured.<br> We did not identify any relationship between dysfunctional high-density lipoprotein and the physical disability, cognitive decline, fatigue and sleep problems of multiple sclerosis. Thiol–disulfide homeostasis was associated with cognitive scores. The shift of the balance towards disulfide was accompanied by a decrease in cognitive scores. On the other hand, we did not detect any relationship between fatigue and sleep disorders and thiol–disulfide homeostasis. Our findings revealed a possible correlation between cognitive dysfunction and thiol–disulfide homeostasis in multiple sclerosis patients. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 1010-6030 452 2024 Photochemical synthesis and solvatochromic fluorescence behavior of imide-fused phenacenes 115613 EN Keito Nose Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Minoru Yamaji Division of Molecular Science, Graduate School of Science and Engineering, Gunma University Fumito Tani Institute for Materials Chemistry and Engineering, Kyushu University Kenta Goto Institute for Materials Chemistry and Engineering, Kyushu University Hideki Okamoto Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Chrysenes, picene, fulminene, modified with imide, bromo, and amino functionalities, were synthesized through Mallory photoreaction as the key step, and their electronic spectra were investigated. Fluorescence spectra of chrysene-diimide CHRDI and bromo-substituted phencanene-imides, BrCHRI, BrPICI, BrFULI were dependent on solvent polarity to display appreciable fluorescence color changes. The solvatofluorochromic behavior was analyzed by conventional relationships between Stokes shift and solvent polarity parameters, such as Lippert-Mataga and Bilot-Kawski equations. The results indicated that the solvatofluorochromism was derived from the intramolecular charge transfer (ICT) nature in the excited state. Theoretical studies using time-dependent density-functional theory revealed that the phenacene-imide molecules in the fluorescent state possessed ICT characters between the strongly electron-withdrawing imide moiety and moderately electron-donating phenacene cores. Amino-substituted chrysene-imide NH2CHRI showed fluorescence band in a red region (λFL = 618 nm) in toluene with a very large Stokes shift (Δ nu= 7630 cm−1) suggesting that the molecule in the fluorescent state was highly polarized. The present results indicate that phenacenes would provide potential platforms for constructing future functional fluorophores through an appropriate functionalization. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0045-6535 358 2024 Size, polyglycerol grafting, and net surface charge of iron oxide nanoparticles determine their interaction and toxicity in Caenorhabditis elegans 142060 EN Yajuan Zou Graduate School of Natural Science and Technology, Okayama University Yutaka Shikano Institute of Systems and Information Engineering, University of Tsukuba Yuta Nishina Graduate School of Natural Science and Technology, Okayama University Naoki Komatsu Graduate School of Human and Environmental Studies, Kyoto University Eriko Kage-Nakadai Department of Nutrition, Graduate School of Human Life and Ecology, Osaka Metropolitan University Masazumi Fujiwara Graduate School of Natural Science and Technology, Okayama University The widespread application of engineered nanoparticles (NPs) in environmental remediation has raised public concerns about their toxicity to aquatic organisms. Although appropriate surface modification can mitigate the ecotoxicity of NPs, the lack of polymer coating to inhibit toxicity completely and the insufficient knowledge about charge effect hinder the development of safe nanomaterials. Herein, we explored the potential of polyglycerol (PG) functionalization in alleviating the environmental risks of NPs. Iron oxide NPs (ION) of 20, 100, and 200 nm sizes (IONS, IONM and IONL, respectively) were grafted with PG to afford ION-PG. We examined the interaction of ION and ION-PG with Caenorhabditis elegans (C. elegans) and found that PG suppressed non-specific interaction of ION with C. elegans to reduce their accumulation and to inhibit their translocation. Particularly, IONS-PG was completely excluded from worms of all developmental stages. By covalently introducing sulfate, carboxyl and amino groups onto IONS-PG, we further demonstrated that positively charged IONS-PG-NH3+ induced high intestinal accumulation, cuticle adhesion and distal translocation, whereas the negatively charged IONS-PG-OSO3– and IONS-PG-COO– were excreted out. Consequently, no apparent deleterious effects on brood size and life span were observed in worms treated by IONS-PG and IONS-PG bearing negatively charged groups. This study presents new surface functionalization approaches for developing ecofriendly nanomaterials. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 78 2 2024 p53-Armed Oncolytic Virotherapy Improves Radiosensitivity in Soft-Tissue Sarcoma by Suppressing BCL-xL Expression 151 161 EN Tadashi Komatsubara Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroshi Tazawa Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Joe Hasei Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshinori Omori Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kazuhisa Sugiu Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yusuke Mochizuki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Koji Demiya Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Aki Yoshida Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomohiro Fujiwara Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiyuki Kunisada Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuo Urata Oncolys BioPharma, Inc. Shunsuke Kagawa Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshifumi Ozaki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiyoshi Fujiwara Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/66924 Soft-tissue sarcoma (STS) is a heterogeneous group of rare tumors originating predominantly from the embryonic mesoderm. Despite the development of combined modalities including radiotherapy, STSs are often refractory to antitumor modalities, and novel strategies that improve the prognosis of STS patients are needed. We previously demonstrated the therapeutic potential of two telomerase-specific replication-competent oncolytic adenoviruses, OBP-301 and tumor suppressor p53-armed OBP-702, in human STS cells. Here, we demonstrate in vitro and in vivo antitumor effects of OBP-702 in combination with ionizing radiation against human STS cells (HT1080, NMS-2, SYO-1). OBP-702 synergistically promoted the antitumor effect of ionizing radiation in the STS cells by suppressing the expression of B-cell lymphoma-X large (BCL-xL) and enhancing ionizing radiation-induced apoptosis. The in vivo experiments demonstrated that this combination therapy significantly suppressed STS tumors’ growth. Our results suggest that OBP-702 is a promising antitumor reagent for promoting the radiosensitivity of STS tumors. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 77 6 2023 Fine Particulate Matter and Diabetes Prevalence in Okayama, Japan 607 612 EN Yasunari Tani Department of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Saori Kashima Environmental Health Sciences Laboratory, Graduate School of Advanced Science and Engineering, Center for the Planetary Health and Innovation Science, The IDEC Institute, Hiroshima University Toshiharu Mitsuhashi Center for Innovate Clinical Medicine, Okayama University Hospital Etsuji Suzuki Department of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Soshi Takao Department of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takashi Yorifuji Department of Epidemiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/66152 Many studies have shown an association between long-term exposure to particulate matter having an aerodynamic diameter of 2.5 μm or less (PM2.5) and diabetes mellitus (DM), but few studies have focused on Asian subjects. We thus examined the association between long-term exposure to PM2.5 and DM prevalence in Okayama City, Japan. We included 76,591 participants who had received basic health checkups in 2006 and 2007. We assigned the census-level modeled PM2.5 data from 2006 and 2007 to each participant and defined DM using treatment status and the blood testing. PM2.5 was associated with DM prevalence, and the prevalence ratio (95% confidence interval) was 1.10 (1.00-1.20) following each interquartile range increase (2.1 μg/m3) in PM2.5. This finding is consistent with previous results and suggests that long-term exposure to PM2.5 is associated with an increased prevalence of DM in Okayama City, Japan, where the PM2.5 level is lower than in other cities in Asian countries. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0889-1575 123 2023 Fluorometric assay of laccase in mushroom extracts and comparisons with absorption spectrophotometry 105627 EN Jianchao Ren Department of Chemistry, Okayama University Kaewta Danchana Department of Chemistry, Okayama University Keiko Sasaki Department of Earth Resources Engineering, Graduate School of Engineering, Kyushu University Takashi Kaneta Department of Chemistry, Okayama University Laccase is a lignin-degrading enzyme that is expected to move industrial applications to a greener form of biotechnology. Here, we used 2,2'-azinobis(3-ethylbenzthiazolin-6-sulfonic acid) (ABTS) as a mediator and N-benzoyl leucomethylene blue (BLMB) as a substrate to develop a fluorometric assay that we used to measure laccase activity in mushroom extracts. We then compared this novel approach to conventional absorption spectrophotometry. With this novel approach, laccase oxidizes ABTS to produce ABTS radicals that show an absorption maximum at 415 nm. The ABTS radicals oxidize BLMB to generate fluorescent methylene blue that is measured by fluorometry while absorption spectrophotometry directly measures the absorbance of the ABTS radicals at 415 nm. Under the optimal conditions, the fluorometric assay showed a linear calibration curve with limits of detection and quantification of 1.0 × 10-2 mg mL-1 and 3.2 × 10-2 mg mL-1, respectively, and those values are 1.4-fold lower than the results using conventional absorption spectrophotometry to measure ABTS radicals. Laccase activity of extracts from species of mushrooms that include eryngii and shiitake were successfully determined via both fluorometry and absorption spectrophotometry. The eryngii extract showed the highest level of activity, which was followed by the shiitake extract, but laccase activity was not observed in the shimeji extract. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 77 5 2023 Association between BRCA Gene Variants and the Response to Modified FOLFIRINOX in Patients with Unresectable Pancreatic Cancer 517 525 EN Shigeru Horiguchi Department of Gastroenterology and Hepatology, Okayama University Hospital Kazuyuki Matsumoto Department of Gastroenterology and Hepatology, Okayama University Hospital Kosaku Morimoto Department of Gastroenterology and Hepatology, Okayama University Hospital Akihiro Matsumi Department of Gastroenterology and Hepatology, Okayama University Hospital Hiroyuki Terasawa Department of Gastroenterology and Hepatology, Okayama University Hospital Yuki Fujii Department of Gastroenterology and Hepatology, Okayama University Hospital Tatsuhiro Yamazaki Department of Gastroenterology and Hepatology, Okayama University Hospital Koichiro Tsutsumi Department of Gastroenterology and Hepatology, Okayama University Hospital Hironari Kato Department of Gastroenterology and Hepatology, Okayama University Hospital Original Article 10.18926/AMO/65974 We investigated the effect of modified FOLFIRINOX (mFFX) in unresectable pancreatic cancer by retrospectively analyzing the cases of 43 patients who underwent BRCA testing (germline, n=11; somatic, n=26; both germline and somatic, n=6). The association between BRCA mutations and therapeutic effect was clarified. Six patients tested positive for germline pathogenic variants. Familial pancreatic cancer (33% vs. 3%, p=0.006) and peritoneal disseminated lesions (66% vs. 8%, p<0.001) were significantly more common in patients with germline pathogenic variants. The partial response (PR) rate was 100% in the germline BRCA-positive patients, and 27% in the germline BRCA-negative patients (p<0.001). The median progression-free survival (PFS) was not reached for any germline BRCA-positive patients but was 9.0 months for the germline BRCA-negative patients (p=0.042). Patients with stage IV BRCA-associated pancreatic cancer had better overall survival than those with non-BRCA-associated pancreatic cancer, although the difference was nonsignificant (not reached vs. 655 days, p=0.061). Our results demonstrate that a PR and prolonged PFS can be expected in germline BRCA-positive patients after treatment with mFFX. Our findings also suggest that germline BRCA pathogenic variants may be useful as biomarkers for the therapeutic effect of mFFX in patients with pancreatic cancer. No potential conflict of interest relevant to this article was reported.

Pharmaceutical Society of Japan Acta Medica Okayama 0009-2363 71 2 2023 Identification of a Functionally Efficient and Thermally Stable Outward Sodium-Pumping Rhodopsin (BeNaR) from a Thermophilic Bacterium 154 164 EN Marie Kurihara Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Vera Thiel Department of Biological Sciences, Tokyo Metropolitan University Hirona Takahashi Department of Chemistry, Graduate School of Science, Okayama University of Science Keiichi Kojima Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University David M. Ward Department of Land Resources and Environmental Sciences, Montana State University Donald A. Bryant Department of Biochemistry and Molecular Biology, The Pennsylvania State University Makoto Sakai Department of Chemistry, Graduate School of Science, Okayama University of Science Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Yuki Sudo Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Rhodopsins are transmembrane proteins with retinal chromophores that are involved in photo-energy conversion and photo-signal transduction in diverse organisms. In this study, we newly identified and characterized a rhodopsin from a thermophilic bacterium, Bellilinea sp. Recombinant Escherichia coli cells expressing the rhodopsin showed light-induced alkalization of the medium only in the presence of sodium ions (Na+), and the alkalization signal was enhanced by addition of a protonophore, indicating an outward Na+ pump function across the cellular membrane. Thus, we named the protein Bellilinea Na+-pumping rhodopsin, BeNaR. Of note, its Na+-pumping activity is significantly greater than that of the known Na+-pumping rhodopsin, KR2. We further characterized its photochemical properties as follows: (i) Visible spectroscopy and HPLC revealed that BeNaR has an absorption maximum at 524 nm with predominantly (>96%) the all-trans retinal conformer. (ii) Time-dependent thermal denaturation experiments revealed that BeNaR showed high thermal stability. (iii) The time-resolved flash-photolysis in the nanosecond to millisecond time domains revealed the presence of four kinetically distinctive photointermediates, K, L, M and O. (iv) Mutational analysis revealed that Asp101, which acts as a counterion, and Asp230 around the retinal were essential for the Na+-pumping activity. From the results, we propose a model for the outward Na+-pumping mechanism of BeNaR. The efficient Na+-pumping activity of BeNaR and its high stability make it a useful model both for ion transporters and optogenetics tools. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0924-4247 361 2023 A soft rotary actuator with a flexible shaft using flexible pneumatic actuators 114603 EN So Shimooka Faculty of Natural Science and Technology, Okayama University Miku Kawanaka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Akio Gofuku Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University This paper proposes a soft rotary actuator that can rotate even when its shaft is bent. The tested rotary actuator consists of three Extension-type Flexible Pneumatic Actuators (EFPA), flexible plates for restraining the EFPAs geometrically, and a polyurethane tube as a shaft. The EFPA consists of a silicone rubber tube covered with a sleeve that can expand significantly in the axial direction when the tube is pressurized. By restraining the EFPA to a helical shape using plates, the proposed rotary actuator can rotate when the three EFPAs are extended in the rotational direction upon the application of pressure. It is confirmed that the tested actuator could rotate even if the shaft is bent, because the shaft and EFPAs consist of flexible materials. The maximum rotation angle and torque are approximately 400° and 0.5 Nm, respectively, for an input pressure of 500 kPa. An analytical model of the tested actuator is proposed to predict the relationship between the rotation angle and the input pressure. A comparison between the calculated and experimental rotation angles reveals that the experimental results can be accurately predicted using the proposed analytical model, which considers the effects of EFPA friction and restraining. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0006-291X 677 2023 Demonstration of iodide-dependent UVA-triggered growth inhibition in Saccharomyces cerevisiae cells and identification of its suppressive molecules 1 5 EN Ryota Ono Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Nozomu Saeki Research Core for Interdisciplinary Sciences, Okayama University Keiichi Kojima Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hisao Moriya Research Core for Interdisciplinary Sciences, Okayama University Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Upon white light illumination, the growth of the budding yeast Saccharomyces cerevisiae was extremely impaired only in the presence of iodide ions, but not fluoride, chloride and bromide ions. Action spectroscopy revealed that the maximum wavelength of the light is around at 373 nm, corresponding to the UVA region. Using a genetic approach, several genes, including OPY1, HEM1, and PAU11, were identified as suppressors of this growth inhibition. This iodide-dependent UVA-triggered growth inhibition method, along with its suppressive molecules, would be beneficial for understanding cell growth processes in eukaryotes and can be utilized for medium sterilization using UVA light. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 77 4 2023 Relationship between the Arthroscopic Findings and Pathology of Greater Trochanteric Pain Syndrome 371 375 EN Yosuke Iwamoto Department of Orthopedic Surgery, Akita University Graduate School of Medicine Mitsunori Kaya Kaya Orthopedic Surgery Sports Clinic Hiroaki Kijima Department of Orthopedic Surgery, Akita University Graduate School of Medicine Masashi Fujii Akita Hip Research Group Itsuki Nagahata Akita Hip Research Group Naohisa Miyakoshi Department of Orthopedic Surgery, Akita University Graduate School of Medicine Original Article 10.18926/AMO/65746 In recent publications on greater trochanteric pain syndrome (GTPS), the pathology receiving the most attention has been gluteus medius muscle tendinous injury, and surgical techniques such as gluteus medius tendon repair and their outcomes for GTPS have been reported. In our department-related facilities, arthroscopic surgeries are routinely performed for the patients with recalcitrant GTPS. A total of 51 patients were diagnosed with GTPS. Surgical treatment was carried out 22 patients (24 joints; 4 males and 18 females; mean age at surgery of 52.0 years). Arthroscopic findings confirmed bursitis in all 24 joints. In all cases, debridement of the greater trochanter bursa provided rapid relief of greater trochanter pain. The Numerical Rating Scale showed significant improvement, from the preoperative mean of 7.8 (range, 6-10) to the postoperative day 7 mean of 1.6 (range, 0-3). The modified Harris Hip Score was significantly improved from the preoperative mean of 65.5 (range, 52.5-78.3) to the final follow-up (average 2.9 months) mean of 96.0 (range, 85.2-100). Fascial damage of the gluteus medius muscle was observed in 21 joints while only 2 patients had a gluteus medius tendinous injury. Greater trochanteric bursitis and fascia or muscle-fiber injury of the gluteus medius muscle are the most common pathologies in patients with lateral hip pain. No potential conflict of interest relevant to this article was reported.

nature portfolio Acta Medica Okayama 2045-2322 13 1 2023 A blue-shifted anion channelrhodopsin from the Colpodellida alga Vitrella brassicaformis 6974 EN Keiichi Kojima Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Shiho Kawanishi Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yosuke Nishimura Research Center for Bioscience and Nanoscience (CeBN), Research Institute for Marine Resources Utilization, Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Masumi Hasegawa Institute for Extra‑Cutting‑Edge Science and Technology Avant‑Garde Research (X‑Star) Shin Nakao Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuya Nagata Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Yuki Sudo Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Microbial rhodopsins, a family of photoreceptive membrane proteins containing the chromophore retinal, show a variety of light-dependent molecular functions. Channelrhodopsins work as light-gated ion channels and are widely utilized for optogenetics, which is a method for controlling neural activities by light. Since two cation channelrhodopsins were identified from the chlorophyte alga Chlamydomonas reinhardtii, recent advances in genomic research have revealed a wide variety of channelrhodopsins including anion channelrhodopsins (ACRs), describing their highly diversified molecular properties (e.g., spectral sensitivity, kinetics and ion selectivity). Here, we report two channelrhodopsin-like rhodopsins from the Colpodellida alga Vitrella brassicaformis, which are phylogenetically distinct from the known channelrhodopsins. Spectroscopic and electrophysiological analyses indicated that these rhodopsins are green- and blue-sensitive pigments (lambda(max) = similar to 550 and similar to 440 nm) that exhibit light-dependent ion channeling activities. Detailed electrophysiological analysis revealed that one of them works as a monovalent anion (Cl-, Br- and NO3-) channel and we named it V. brassicaformis anion channelrhodopsin-2, VbACR2. Importantly, the absorption maximum of VbACR2 (similar to 440 nm) is blue-shifted among the known ACRs. Thus, we identified the new blue-shifted ACR, which leads to the expansion of the molecular diversity of ACRs. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0032-3896 2023 Preparation of crystalline polyimide nanofibers via solution crystallization EN Shota Kumano Graduate School of Natural Science and Technology, Okayama University Tomoyasu Takaki Graduate School of Natural Science and Technology, Okayama University Tetsuya Uchida Graduate School of Natural Science and Technology, Okayama University Two crystalline polyimide nanofibers (PINFs) with different morphologies were prepared. The crystalline unit cells of the aromatic PI crystals and the crystal morphologies of the fabricated PINFs were examined. PINF-I (lengths = 305 ± 152 nm and diameters = 12 ± 2 nm) was crystallized from crystalline PI dissolved in a concentrated sulfuric acid solution. The resulting PINF-I was isolated from this solution, and it did not aggregate in water. PINF-II with diameters of 105 ± 99 nm was prepared by dispersing PINF-I in a mixed water and t-butanol (TBA) solution (water:TBA = 4:1), followed by freeze-drying. Then, the PINF-II was heated to enhance its crystallinity. X-ray diffraction and transmission electron microscopy studies of the heat-treated PINF-II revealed a PI crystalline unit cell [orthorhombic, a = 1.21 nm, b = 0.88 nm, and c = 2.23 nm (molecular chain axis direction)]. The crystal structure of the heat-treated PINF-II suggested that highly crystalline PINFs were fabricated in which the PI molecular chains were oriented along the direction of the fiber lengths. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2072-6694 15 3 2023 Therapeutic Strategies to Overcome Fibrotic Barriers to Nanomedicine in the Pancreatic Tumor Microenvironment 724 EN Hiroyoshi Y. Tanaka Department of Pharmaceutical Biomedicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takuya Nakazawa Department of Pharmaceutical Biomedicine, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Atsushi Enomoto Department of Pathology, Graduate School of Medicine, Nagoya University Atsushi Masamune Division of Gastroenterology, Graduate School of Medicine, Tohoku University Mitsunobu R. Kano Department of Pharmaceutical Biomedicine, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Simple Summary Pancreatic cancer is difficult to treat. Novel treatment strategies are urgently needed to improve the survival rate, which is approximately 10% five years after diagnosis. The use of nanomedicines, which are formulated within a characteristic size range that favors its specific delivery to the diseased tissue, is being actively explored in cancer treatment. However, fibrosis (the abnormal accumulation of a cell type called fibroblasts and the fibrous protein network that they create) is characteristically seen in pancreatic cancer and hinders the delivery of nanomedicines into cancerous tissue. The decreased efficiency of delivery limits the therapeutic effects of nanomedicine in pancreatic cancer. We call this the "fibrotic barrier" to nanomedicine. To overcome the fibrotic barrier, we could target the fibrotic process and/or optimize the nanomedicine design. In this review, we give a detailed overview of strategies to overcome the fibrotic barriers in pancreatic cancer and highlight key gaps in our understanding. Pancreatic cancer is notorious for its dismal prognosis. The enhanced permeability and retention (EPR) effect theory posits that nanomedicines (therapeutics in the size range of approximately 10-200 nm) selectively accumulate in tumors. Nanomedicine has thus been suggested to be the "magic bullet"-both effective and safe-to treat pancreatic cancer. However, the densely fibrotic tumor microenvironment of pancreatic cancer impedes nanomedicine delivery. The EPR effect is thus insufficient to achieve a significant therapeutic effect. Intratumoral fibrosis is chiefly driven by aberrantly activated fibroblasts and the extracellular matrix (ECM) components secreted. Fibroblast and ECM abnormalities offer various potential targets for therapeutic intervention. In this review, we detail the diverse strategies being tested to overcome the fibrotic barriers to nanomedicine in pancreatic cancer. Strategies that target the fibrotic tissue/process are discussed first, which are followed by strategies to optimize nanomedicine design. We provide an overview of how a deeper understanding, increasingly at single-cell resolution, of fibroblast biology is revealing the complex role of the fibrotic stroma in pancreatic cancer pathogenesis and consider the therapeutic implications. Finally, we discuss critical gaps in our understanding and how we might better formulate strategies to successfully overcome the fibrotic barriers in pancreatic cancer. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 77 1 2023 Prevalence of Inducible Macrolide, Lincosamide, and Streptogramin B (inducible MLSB) Resistance in Clindamycin-Susceptible Staphylococcus aureus at Okayama University Hospital 1 9 EN Lutfun Nahar Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hideharu Hagiya Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takahiro Nada Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Koji Iio Microbiology Division, Clinical Laboratory, Okayama University Hospital Kazuyoshi Gotoh Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Osamu Matsushita Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Fumio Otsuka Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/64355 Inducible resistance to the macrolide, lincosamide, and streptogramin B (iMLSB) antibiotic family is a latent mechanism for antimicrobial resistance in Staphylococcus aureus. We here investigated the frequency and genotypic profiles of iMLSB resistance in clindamycin (CLDM)-susceptible S. aureus isolated in Okayama University Hospital from June 2020 to June 2021. We phenotypically screened the iMLSB resistance via D-zone test and performed PCR testing for the erythromycin ribosomal methylase (erm) genes: ermA and ermC. Among 432 CLDM-susceptible S. aureus isolates, 138 (31.9%) exhibited an iMLSB-resistance phenotype, with methicillinresistant S. aureus isolates (MRSA; 61 isolates: 58.6%) exhibiting higher positivity than methicillin-sensitive S. aureus isolates (MSSA; 77 isolates: 23.5%) (p<0.001). Male patients had a higher frequency of iMLSB resistance than females (OR [95%CI]: 1.8 [1.2-2.8]; p=0.007). Genotypically, ermA predominated in both MSSA (70.1%) and MRSA (86.9%) compared to ermC (14.3% in MSSA and 11.5% in MRSA). A single strain of MRSA possessed both ermA and ermC, while 12 (15.6%) MSSA isolates were negative for both ermA and ermC, suggesting the presence of other genetic mechanisms. Collectively, these results show that approximately 33% of CLDM-susceptible S. aureus isolates at our university hospital exhibited iMLSB resistance, predominantly caused by ermA in both MSSA and MRSA. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 6 2022 A Rare Case of Idiopathic Spinal Cord Herniation Treated by DuraGen® Collagen Matrix Graft 731 736 EN Maho Kamamura Department of Radiology, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Fumiyo Higaki Department of Radiology, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Susumu Sasada Department of Neurological Surgery, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Toshi Matsushita Division of Radiological Technology, Okayama University Hospital Takao Yasuhara Department of Neurological Surgery, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Isao Date Department of Neurological Surgery, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Takao Hiraki Department of Radiology, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Case Report 10.18926/AMO/64124 We report a rare case of idiopathic spinal cord herniation (ISCH) with a history of cerebrospinal fluid (CSF) leakage. ISCH is a protrusion of the spinal cord through a dural defect. Thin constructive interference in steady-state (CISS) images clearly demonstrated the herniated cord in the present case. The myelopathy worsened and the patient underwent surgery for reduction of herniated spinal cord; the dural defect was filled by placing collagen matrix graft (DuraGen^®) between the inner and outer dural layers. The patient’s symptoms have improved without relapse for 8 months since surgery. This method may be a good surgical option for cases of spinal cord herniation. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 6 2022 Handling of Germline Findings in Clinical Comprehensive Cancer Genomic Profiling 673 678 EN Mika Okazawa-Sakai Department of Clinical Genomic Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuko Yamamoto Department of Cancer Genomic Medicine, National Hospital Organization Shikoku Cancer Center Mashu Futagawa Department of Clinical Genomic Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Miki Okamura Department of Hereditary Tumors, National Hospital Organization Shikoku Cancer Center Satoko Miyawaki Department of Cancer Genomic Medicine, National Hospital Organization Shikoku Cancer Center Tomohiro Nishina Department of Cancer Genomic Medicine, National Hospital Organization Shikoku Cancer Center Kazuhiro Takehara Department of Gynecologic Oncology, National Hospital Organization Shikoku Cancer Center Toshiyuki Kozuki Department of Clinical Research Center, National Hospital Organization Shikoku Cancer Center Shuta Tomida Center for Comprehensive Genomic Medicine, Okayama University Hospital Ichinosuke Hyodo Department of Cancer Genomic Medicine, National Hospital Organization Shikoku Cancer Center Shozo Ohsumi Department of Hereditary Tumors, National Hospital Organization Shikoku Cancer Center Akira Hirasawa Department of Clinical Genomic Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/64117 Patients found to have presumed germline pathogenic variants (PGPVs) during comprehensive genomic profiling (CGP) require genetic counseling (GC) referrals. We retrospectively investigated the outcomes of patients with PGPVs. Among 159 patients who underwent CGP, we recommended GC for the 16 patients with PGPVs (3 with [FG group] and 13 without [G Group] a family/personal history of hereditary cancer) as well as for the 8 patients with no PGPVs, but a history (F group); 2 (67%), 5 (38%), and 3 (38%) patients received GC in the FG, G, and F groups, respectively. Germline testing results were positive in 1 and 2 patients of the FG and G groups, respectively. Among the patients recommended for GC, 58% did not receive GC due to lack of interest, poor performance status, or death. CGP contributes to the identification of germline variants in patients without a history of hereditary cancer. However, the proportion of patients who undergo GC should be improved. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 5 2022 Surgery Outcomes for Pulmonary Metastases from Renal Cell Carcinoma 585 591 EN Haruki Choshi Department of Thoracic Surgery, National Hospital Organization Iwakuni Clinical Center Mototsugu Watanabe Department of Thoracic Surgery, National Hospital Organization Iwakuni Clinical Center Shinichi Furukawa Department of Thoracic Surgery, National Hospital Organization Iwakuni Clinical Center Hiroyuki Ujike Department of Thoracic Surgery, National Hospital Organization Iwakuni Clinical Center Kazuhiko Kataoka Department of Thoracic Surgery, National Hospital Organization Iwakuni Clinical Center Original Article 10.18926/AMO/64040 Pulmonary metastatic resection is a standard therapy for renal cell carcinoma (RCC). Although patients with pulmonary metastases who do not undergo any treatment have poor prognoses, it has been reported that resection for pulmonary metastases yields good clinical outcomes. We investigated the prognoses of the 10 Japanese patients (eight males, two females) who underwent a surgical resection of pulmonary metastasectomy from RCC at our institution between April 1, 2012 and March 31, 2020 and analyzed the prognostic factors. We determined the prognoses and calculated the 5-year overall survival (OS) and disease-free survival (DFS) rates. To identify prognostic factors, we compared the median DFS duration for each factor. Elderly patients (median age, 75.5 years) were more predominant compared to previous studies, and all 10 patients underwent a complete resection. The 5-year DFS rate was 30.5% (95%CI: 0.045-0.63) and the 5-year OS rate was 80% (95%CI: 0.20-0.97). The following factors were associated with better prognosis: female, disease-free interval≥36 months, and metastases size<12 mm. These results indicate that complete resection for pulmonary metastases from RCC resulted in good clinical outcomes, particularly for patients with better prognostic factors. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 5 2022 Relationship between Wearing a Lead Apron and Work-related Musculoskeletal Disorders: A Questionnaire Survey of Japanese Radiological Technologists 541 545 EN Toru Akebi Department of Physical Therapy, Faculty of Health Sciences, Okayama Healthcare Professional University Ryutaro Matsugaki Department of Preventive Medicine and Community Health, School of Medicine, University of Occupational and Environmental Health Toshiro Ono Department of Physical Therapy, Faculty of Health Sciences, Okayama Healthcare Professional University Original Article 10.18926/AMO/64035 The purpose of this study was to conduct a self-reported questionnaire survey of work-related musculoskeletal disorders (WMSDs) among Japanese radiological technologists (RTs) and to report on the relationship between wearing a lead apron and WMSDs. Between February and April of 2021, RTs in Okayama Prefecture, Japan, were surveyed by mail and through a website. Information on individual characteristics, physical factors at work, and the presence of WMSDs were collected. All participants were also asked whether they frequently wore lead aprons. A multiple logistic regression analysis was used to assess the relationship between wearing a lead apron and WMSDs. The model was adjusted for age, sex, body mass index (BMI), and working hours. Of the 123 participants, 67 (54.5%) had WMSDs. Multiple logistic regression analysis revealed that WMSDs were significantly associated with wearing a lead apron. Compared to the “Never wear” group, the odds ratios for the “Always/Frequently wear” and “Sometimes/Rarely wear” groups were 7.87 (95% confidence interval [CI]=1.28-48.46; p=0.026) and 7.80 (95% CI=1.43-42.44; p=0.017), respectively. Our analysis suggests that wearing a lead apron is associated with WMSDs, and thus design modifications in lead aprons may improve the occupational health management of RTs. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 5 2022 Viral Sequences Are Repurposed for Controlling Antiviral Responses as Non-Retroviral Endogenous Viral Elements 503 510 EN Hirohito Ogawa Department of Virology, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Tomoyuki Honda Department of Virology, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Review 10.18926/AMO/64025 Eukaryotic genomes contain numerous copies of endogenous viral elements (EVEs), most of which are considered endogenous retrovirus (ERV) sequences. Over the past decade, non-retroviral endogenous viral elements (nrEVEs) derived from ancient RNA viruses have been discovered. Several functions have been proposed for these elements, including antiviral defense. This review summarizes the current understanding of nrEVEs derived from RNA viruses, particularly endogenous bornavirus-like elements (EBLs) and endogenous filovirus-like elements (EFLs). EBLs are one of the most extensively studied nrEVEs. The EBL derived from bornavirus nucleoprotein (EBLN) is thought to function as a non-coding RNA or protein that regulates host gene expression or inhibits virus propagation. Ebolavirus and marburgvirus, which are filoviruses, induce severe hemorrhagic fever in humans and nonhuman primates. Although the ecology of filoviruses remains unclear, bats are believed to be potential reservoirs. Based on the knowledge from EBLs, it is postulated that EFLs in the bat genome help to maintain the balance between filovirus infection and the bat’s defense system, which may partially explain why bats act as potential reservoirs. Further research into the functions of nrEVEs could reveal novel antiviral systems and inspire novel antiviral approaches. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 4 2022 Molecular-targeted Therapy for Metastatic Renal Cell Carcinoma As First-line Therapy: A Single Institution 13-year Experience 465 472 EN Kensuke Bekku Department of Urology, Hiroshima City Hiroshima Citizens Hospital Takuji Tsugawa Department of Urology, Hiroshima City Hiroshima Citizens Hospital Kazuma Tsuboi Department of Urology, Hiroshima City Hiroshima Citizens Hospital Gaku Noda Department of Urology, Hiroshima City Hiroshima Citizens Hospital Yousuke Inoue Department of Urology, Hiroshima City Hiroshima Citizens Hospital Wataru Murao Department of Urology, Hiroshima City Hiroshima Citizens Hospital Shin Ebara Department of Urology, Hiroshima City Hiroshima Citizens Hospital Original Article 10.18926/AMO/63906 We aimed to identify the role of first-line monotherapy with vascular endothelial growth factor receptor tyrosine kinase inhibitors (VEGFR-TKI) in patients with metastatic RCC. Eligible patients were categorized into three groups (favorable, intermediate, and poor risk) according to the International Metastatic RCC Database Consortium risk criteria. Overall survival (OS) was the primary endpoint. Survival was compared using the log-rank test. A total of 108 patients were retrospectively analyzed. The numbers of patients in the favorable-, intermediate-, and poor-risk groups were 32 (30%), 66 (61%), and 10 (9%), repestively. The median OS values in the entire cohort was 36 months (95% confidence interval [CI] 29-53). The median OS in the favorable, intermediate, and poor risk groups were 94 months (95% CI: 43-Not reached), 30 months (95% CI: 20-38), and 8 months (95% CI: 0-Not reached), respectively (p<0.05). Prior nephrectomy, clear cell histology, clinical T stage ≤2, no metastasis at the time of diagnosis, nivolumab beyond first-line therapy, and objective response to VEGFR-TKIs were factors significantly prolonging OS on univariate analysis. VEGFR-TKI monotherapy as first-line therapy was an effective treatment option for patients with metastatic clear cell RCC with favorable risk. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 4 2022 Effects of Exercise Therapy and Nutrition Therapy on Patients with Possible Malnutrition and Sarcopenia in a Recovery Rehabilitation Ward 423 428 EN Satoshi Takahashi Department of Rehabilitation Medicine, Saiseikai Imabari Daini Hospital Takuya Kushibe Department of Rehabilitation Medicine, Saiseikai Imabari Daini Hospital Yoshiteru Akezaki Division of Physical Therapy, Kochi Professional University of Rehabilitation Norio Horiike Department of Internal Medicine, Saiseikai Imabari Daini Hospital Original Article 10.18926/AMO/63901 We compared the effects of an exercise intervention with that of exercise combined with nutrition therapy in patients with possible malnutrition and sarcopenia admitted to a recovery rehabilitation ward, and we examined the differences in the patients’ physical function and activities of daily living (ADLs). There were 16 patients in the Exercise group with exercise therapy and ADL exercises, and 14 patients in the Combined intervention group with exercise therapy, ADL exercises, and nutrition therapy. The survey items were body weight, body mass index, grip strength, lower-leg circumference, gait speed, and ADLs, each of which was measured at the baseline and at 2 weeks, 4 weeks, and at discharge. Significant improvements in grip strength were observed in the Combined intervention group as follows: at 4 weeks>at 2 weeks (p<0.05), and at discharge>baseline and 2 weeks (p<0.05). There were no significant changes in the Exercise group, and an interaction was recognized in both groups. Comprehensive rehabilitation including nutrition therapy is necessary for patients with possible malnutrition and/or sarcopenia, as our results indicate that nutrition therapy in addition to exercise therapy has the effect of promoting improvements of physical function in such patients. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 4 2022 Gene Expression Profiling between Patient Groups with High and Low Ki67 Levels after Short-term Preoperative Aromatase Inhibitor Treatment for Breast Cancer 399 408 EN Yukiko Kajiwara Department of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takayuki Iwamoto Departments of Breast and Endocrine Surgery, Okayama University Hospital Yidan Zhu Department of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mariko Kochi Departments of Breast and Endocrine Surgery, Okayama University Hospital Tadahiko Shien Departments of Breast and Endocrine Surgery, Okayama University Hospital Naruto Taira Departments of Breast and Endocrine Surgery, Okayama University Hospital Hiroyoshi Doihara Departments of Breast and Endocrine Surgery, Okayama University Hospital Shinichi Toyooka Department of General Thoracic Surgery and Breast and Endocrinological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/63894 According to a recent report, a low Ki67 level after short-term preoperative hormone therapy (post-Ki67) might suggest a more favorable prognosis compared with a high post-Ki67 level in patients with hormone receptorpositive/human epidermal growth factor 2-negative (HR+/HER2−) breast cancer with high levels of Ki67. This study aimed to evaluate the pre-treatment genetic differences between these two patient groups. Forty-five luminal B-like patients were stratified into two groups, namely, a group with high (H→H) and one with low (H→L) Ki67 levels after short-term preoperative aromatase inhibitor (AI) treatment. We compared pre-treatmentgene expression profiles between the two groups. In gene level analysis, there was no significant difference between the two groups by the class comparison test. In pathway analysis, five metabolism-related gene sets were significantly upregulated in the H→L group (p≤0.05). In the search for novel targets, five genes (PARP, BRCA2, FLT4, CDK6, and PDCD1LG2) showed significantly higher expression in the H→H group (p≤0.05). Several metabolism-related pathways were associated with sensitivity to AI. In the future, it will be necessary to seek out new therapeutic strategies for the poor prognostic group with high post-Ki67. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1999-4915 14 8 2022 A Transfectable Fusagravirus from a Japanese Strain of Cryphonectria carpinicola with Spherical Particles 1722 EN Subha Das Institute of Plant Science and Resources, Okayama University Sakae Hisano Institute of Plant Science and Resources, Okayama University Ana Eusebio-Cope Institute of Plant Science and Resources, Okayama University Hideki Kondo Institute of Plant Science and Resources, Okayama University Nobuhiro Suzuki Institute of Plant Science and Resources, Okayama University A novel dsRNA virus (Cryphonectria carpinicola fusagravirus 1, CcFGV1), isolated from a Japanese strain (JS13) of Cryphonectria carpinicola, was thoroughly characterized. The biological comparison of a set of isogenic CcFGV1-infected and -free (JS13VF) strains indicated asymptomatic infection by CcFGV1. The sequence analysis showed that the virus has a two open reading frame (ORF) genome of 9.6 kbp with the RNA-directed RNA polymerase domain encoded by ORF2. The N-terminal sequencing and peptide mass fingerprinting showed an N-terminally processed or degraded product (150 kDa) of the 5'-proximal ORF1-encoded protein (1462 amino acids) to make up the CcFGV1 spherical particles of similar to 40 nm in diameter. Interestingly, a portion of CcFGV1 dsRNA co-fractionated with a host protein of 70 kDa. The purified CcFGV1 particles were used to transfect protoplasts of JS13VF as well as the standard strain of an experimental model filamentous fungal host Cryphonectria parasitica. CcFGV1 was confirmed to be associated with asymptomatic infection of both fungi. RNA silencing was shown to target the virus in C. parasitica, resulting in reduced CcFGV1 accumulation by comparing the CcFGV1 content between RNA silencing-competent and -deficient strains. These results indicate the transfectability of spherical particles of a fusagravirus associated with asymptomatic infection. No potential conflict of interest relevant to this article was reported.

American Chemical Society Acta Medica Okayama 2470-1343 2022 Characteristics of Vertical Ga2O3 Schottky Junctions with the Interfacial Hexagonal Boron Nitride Film 26021 26028 EN Venkata Krishna Rao Rama Graduate School of Natural Science and Technology, Okayama University Ajinkya K. Ranade Department of Physical Science and Engineering, Nagoya Institute of Technology Pradeep Desai Department of Physical Science and Engineering, Nagoya Institute of Technology Bhagyashri Todankar Department of Physical Science and Engineering, Nagoya Institute of Technology Golap Kalita Department of Physical Science and Engineering, Nagoya Institute of Technology Hiroo Suzuki Graduate School of Natural Science and Technology Masaki Tanemura Department of Physical Science and Engineering, Nagoya Institute of Technology Yasuhiko Hayashi We present the device properties of a nickel (Ni)- gallium oxide (Ga2O3) Schottky junction with an interfacial hexagonal boron nitride (hBN) layer. A vertical Schottky junction with the configuration Ni/hBN/Ga2O3/In was created using a chemical vapor-deposited hBN film on a Ga(2)O(3 )substrate. The current-voltage characteristics of the Schottky junction were investigated with and without the hBN interfacial layer. We observed that the turn-on voltage for the forward current of the Schottky junction was significantly enhanced with the hBN interfacial film. Furthermore, the Schottky junction was analyzed under the illumination of deep ultraviolet light (254 nm), obtaining a photoresponsivity of 95.11 mA/W under an applied bias voltage (-7.2 V). The hBN interfacial layer for the Ga2O3-based Schottky junction can serve as a barrier layer to control the turn-on voltage and optimize the device properties for deep-UV photosensor applications. Furthermore, the demonstrated vertical heterojunction with an hBN layer has the potential to be significant for temperature management at the junction interface to develop reliable Ga2O3-based Schottky junction devices. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 3 2022 Sequential Development of Putaminal Hemorrhage and Corona Radiata Infarction in the Same Lenticulostriate Arterial Territory 329 332 EN Yuki Inomata Department of Neurosurgery, Shinshu University School of Medicine Yoshiki Hanaoka Department of Neurosurgery, Shinshu University School of Medicine Yu Fujii Department of Neurosurgery, Shinshu University School of Medicine Toshihiro Ogiwara Department of Neurosurgery, Shinshu University School of Medicine Tetsuyoshi Horiuchi Department of Neurosurgery, Shinshu University School of Medicine Case Report 10.18926/AMO/63743 Putaminal hemorrhage is a common type of spontaneous cerebral hemorrhage. However, to our knowledge, there have been no reports of sequential cerebral hemorrhage and infarction in the same perforating arterial territory. Herein, we present the first reported case of the sequential development of putaminal hemorrhage and corona radiata infarction in the same lenticulostriate arterial (LSA) territory. Early intensive blood pressure lowering treatment may have aggravated ischemic damage. If a patient presents with motor weakness that cannot be explained by putaminal hemorrhage, the sequential development of corona radiata infarction in the same LSA territory should be ruled out. No potential conflict of interest relevant to this article was reported.

Institute of Electrical and Electronics Engineers (IEEE) Acta Medica Okayama 0093-9994 58 3 2022 SMC Development Guidelines for Axial Flux PM Machines Employing Coreless Rotor Structure for Enhancing Efficiency Based on Experimental Results 3470 3485 EN Ren Tsunata Division of Industrial Innovation Sciences Graduate School of Natural Science and Technology, Okayama University Masatsugu Takemoto Division of Industrial Innovation Sciences Graduate School of Natural Science and Technology, Okayama University Satoshi Ogasawara Graduate School of Information Science and Technology, Hokkaido University Tatsuya Saito Sumitomo Electric Industries Ltd. Tomoyuki Ueno Sumitomo Electric Industries Ltd. In recent years, there has been growing demand for flat electrical machines in order to minimize system size while also maintaining high efficiency. In general, axial flux machines (AFMs) are more suitable for flat form than radial flux machines (RFMs). AFMs usually employ a soft magnetic composite (SMC) for the stator core, and their efficiency can be improved by employing an SMC that has low iron loss. However, this reduces the average torque because there is generally a tradeoff relationship between iron loss and magnetic permeability in SMCs. This article, therefore, proposes an AFM with a coreless rotor structure in which the torque performance is not easily affected by the permeability of the SMC. This article aims to reveal the ideal SMC characteristics for high efficiency in AFMs. First, many virtual SMC materials with different iron loss and permeability are used for simulations of AFMs in order to investigate sensitivity to material properties. The simulations by virtual SMCs take into account the experimental increase in the iron loss. As a result, guidelines for developing SMCs are constructed based on the experimental results. Finally, prototype AFMs employing new SMCs developed by following the guidelines are shown, and it is found that they offer higher efficiency in all operating area than conventional AFMs. In particular, one prototype achieves an extremely high efficiency of over 96% at 6000 r/min, 0.8 Nm in experiments. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 2 2022 The Combination of D-dimer and Glasgow Prognostic Score Can Be Useful in Predicting VTE in Patients with Stage IIIC and IVA Ovarian Cancer 129 135 EN Kotaro Kubo Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Keiichiro Nakamura Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kazuhiro Okamoto Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hirofumi Matsuoka Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Naoyuki Ida Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomoko Haruma Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Chikako Ogawa Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hisashi Masuyama Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/63406 Cancer patients have increased risk of venous thromboembolism (VTE) that must be assessed before treatment. This study aimed to determine effective VTE biomarkers in gynecologic cancer (GC). We investigated the correlation between D-dimer levels, Khorana risk score (KRS), Glasgow prognostic score (GPS), and VTE in 1499 GC patients (583 cervical cancer (CC), 621 endometrial cancer (EC), and 295 ovarian cancer (OC) patients) treated at our institution between January 2008 and December 2019. χ2 and Mann–Whitney U-tests were used to determine statistical significance. We used receiver operating characteristic-curve analysis to evaluate the discriminatory ability of each parameter. D-dimer levels were significantly correlated with KRS and GPS in patients with GC. VTE was diagnosed in 11 CC (1.9%), 27 EC (4.3%), and 39 OC patients (13.2%). Optimal D-dimer cut-off values for VTE were 3.1, 3.2, and 3.9 μg/ml in CC, EC and OC patients, respectively. D-dimer could significantly predict VTE in all GC patients. Furthermore, D-dimer combined with GPS was more accurate in predicting VTE than other VTE biomarkers in stage IIIC and IVA OC (AUC: 0.846; p<0.001). This study demonstrates that combined D-dimer and GPS are useful in predicting VTE in patients with OC. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1735-1472 2022 Fuel spray impingement and liquid film formation in a gasoline direct-injection spark-ignition engine EN Yasutaka Tomomatsu Graduate School of Natural Science and Technology, Okayama University Nobuyuki Kawahara Graduate School of Natural Science and Technology, Okayama University Eiji Tomita Graduate School of Natural Science and Technology, Okayama University It is important to improve the thermal efficiency and reduce the harmful exhaust emissions of the direct-injection spark-ignition engine. However, this engine has problems such as the emission of particulate matter, including soot, from pool fire with luminous flames. Pool fire is caused by the thermal decomposition of a liquid film, which is created by fuel spray impinging on a piston surface. An understanding of liquid film formation process is important to reduce particulate matter. The purpose of this investigation was to evaluate the effects of injection pressure on fuel spray impingement and liquid film formation process, under engine motoring conditions, using the laser-induced fluorescence method. The fuel consisted of isooctane, 1-octanol and rhodamine B. 1-Octanol was the solvent for rhodamine B, which was illuminated with a neodymium-doped yttrium aluminum garnet laser, causing it to emit red fluorescence at a wavelength of 580 nm; the second harmonic of the laser is at 532 nm. Liquid film images were captured using a high-speed camera. Using image processing, the liquid film area, thickness and mass were estimated. It was found that increasing injection pressure increased the liquid film area, thinned the film and decreased the mass of fuel that remained. In total, 35.6% and 32.5% of the injection mass remained on the piston surface at an injection pressure of 5 and 13 MPa, respectively. In addition, the in-cylinder flow affected the liquid film formation process, stretching the film in the direction of the flow. No potential conflict of interest relevant to this article was reported.

IOP Publishing Ltd Acta Medica Okayama 2631-8644 4 2 2022 High-quality micro-shape fabrication of monocrystalline diamond by nanosecond pulsed laser and acid cleaning 025301 EN Yasuhiro Okamoto Graduate School of Natural Science and Technology, Okayama University Tubasa Okubo Graduate School of Natural Science and Technology, Okayama University Atsuya Kajitani Graduate School of Natural Science and Technology, Okayama University Akira Okada Graduate School of Natural Science and Technology, Okayama University The flat plane of small surface roughness below 0.1 mu m average roughness was obtained for monocrystalline diamond by nanosecond pulsed laser irradiation of 1060 nm and post-process acid cleaning, at a laser fluence around the material removal threshold value. The glossy and flat plane at the bottom of the micro-groove was parallel to the top surface of the specimen, although the round beam of Gaussian mode was irradiated in the direction perpendicular to the top surface of specimen. The square beam of top-hat mode produced a shallower micro-groove with a wider, flatter bottom compared with the round beam in Gaussian mode. The creation method of the flat plane with small surface roughness was discussed in the arrangement strategy of linear micro-grooving by the square beam of top-hat mode. Normal side-by-side repetition of linear micro-grooving did not create a flat plane with constant depth. Therefore, a two-step scanning method was proposed in order to overcome the problem in the normal side-by-side repetition of liner micro-grooving. Non-removal areas were partly retained between the processing lines in the first step, and the laser scanning was conducted on the retained area in the second step. The newly proposed two-step scanning method was practical and useful to create a widely flat plane with small surface roughness, and the two-step scanning method provided superior control over the micro-groove depth. This proposed method can reduce the surface roughness in addition to the shape creation of monocrystalline diamond, and it can be used as a high-quality micro-shape fabrication method of monocrystalline diamond. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0304-8608 167 2022 A novel victorivirus from the phytopathogenic fungus Neofusicoccum parvum 923 929 EN Haris Ahmed Khan Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST) Yukiyo Sato Institute of Plant Science and Resources, Okayama University Hideki Kondo Institute of Plant Science and Resources, Okayama University Atif Jamal Crop Diseases Research Institute, National Agricultural Research Centre Muhammad Faraz Bhatti Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST) Nobuhiro Suzuki Institute of Plant Science and Resources, Okayama University Neofusicoccum parvum is an important plant-pathogenic ascomycetous fungus that causes trunk diseases in a variety of plants. A limited number of reports on mycoviruses from this fungus are available. Here, we report the characterization of a novel victorivirus, Neofusicoccum parvum victorivirus 3 (NpVV3). An agarose gel dsRNA profile of a Pakistani strain of N. parvum, NFN, showed a band of similar to 5 kbp that was not detectable in Japanese strains of N. parvum. Taking a high-throughput and Sanger sequencing approach, the complete genome sequence of NpVV3 was determined to be 5226 bp in length with two open reading frames (ORF1 and ORF2) that encode a capsid protein (CP) and an RNA-dependent RNA polymerase (RdRP). The RdRP appears to be translated by a stop/restart mechanism facilitated by the junction sequence AUGucUGA, as is found in some other victoriviruses. BLASTp searches showed that NpVV3 CP and RdRP share the highest amino acid sequence identity (80.5% and 72.4%, respectively) with the corresponding proteins of NpVV1 isolated from a French strain of N. parvum. However, NpVV3 was found to be different from NpVV1 in its terminal sequences and the stop/restart facilitator sequence. NpVV3 particles similar to 35 nm in diameter were partially purified and used to infect an antiviral-RNA-silencing-deficient strain (Delta cl2) of an experimental ascomycetous fungal host, Cryphonectria parasitica. NpVV3 showed symptomless infection in the new host strain. No potential conflict of interest relevant to this article was reported.

Frontiers Media SA Acta Medica Okayama 2296-889X 8 2021 Exploring the Retinal Binding Cavity of Archaerhodopsin-3 by Replacing the Retinal Chromophore With a Dimethyl Phenylated Derivative 794948 EN Taichi Tsuneishi Laboratory of Biophysical Chemistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Masataka Takahashi Laboratory of Synthetic and Medicinal Chemistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Masaki Tsujimura Department of Applied Chemistry, The University of Tokyo Keiichi Kojima Laboratory of Biophysical Chemistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroshi Ishikita Department of Applied Chemistry, The University of Tokyo Yasuo Takeuchi Laboratory of Synthetic and Medicinal Chemistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuki Sudo Laboratory of Biophysical Chemistry, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Rhodopsins act as photoreceptors with their chromophore retinal (vitamin-A aldehyde) and they regulate light-dependent biological functions. Archaerhodopsin-3 (AR3) is an outward proton pump that has been widely utilized as a tool for optogenetics, a method for controlling cellular activity by light. To characterize the retinal binding cavity of AR3, we synthesized a dimethyl phenylated retinal derivative, (2E,4E,6E,8E)-9-(2,6-Dimethylphenyl)-3,7-dimethylnona-2,4,6,8-tetraenal (DMP-retinal). QM/MM calculations suggested that DMP-retinal can be incorporated into the opsin of AR3 (archaeopsin-3, AO3). Thus, we introduced DMP-retinal into AO3 to obtain the non-natural holoprotein (AO3-DMP) and compared some molecular properties with those of AO3 with the natural A1-retinal (AO3-A1) or AR3. Light-induced pH change measurements revealed that AO3-DMP maintained slow outward proton pumping. Noteworthy, AO3-DMP had several significant changes in its molecular properties compared with AO3-A1 as follows; 1) spectroscopic measurements revealed that the absorption maximum was shifted from 556 to 508 nm and QM/MM calculations showed that the blue-shift was due to the significant increase in the HOMO-LUMO energy gap of the chromophore with the contribution of some residues around the chromophore, 2) time-resolved spectroscopic measurements revealed the photocycling rate was significantly decreased, and 3) kinetical spectroscopic measurements revealed the sensitivity of the chromophore binding Schiff base to attack by hydroxylamine was significantly increased. The QM/MM calculations show that a cavity space is present at the aromatic ring moiety in the AO3-DMP structure whereas it is absent at the corresponding beta-ionone ring moiety in the AO3-A1 structure. We discuss these alterations of the difference in interaction between the natural A1-retinal and the DMP-retinal with binding cavity residues. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 6 2021 EG-VEGF Induces Invasion of a Human Trophoblast Cell Line via PROKR2 677 684 EN Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takashi Mitsui Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Sakurako Mishima Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Akiko Ohira Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Jota Maki Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Eriko Eto Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kei Hayata Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Keiichiro Nakamura Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hisashi Masuyama Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/62806 Extravillous trophoblast (EVT) invasion is important for embryo implantation, placental development, and successful remodeling of the uterine spiral artery. Endocrine gland derived-vascular endothelial growth factor (EG-VEGF) and matrix metalloproteinases (MMPs) are implicated in EVT invasion; however, the high con-centrations found in pregnancy pathologies have not been investigated in non-tumor trophoblasts. The roles of EG-VEGF, prokineticin receptors (PROKR1/2), MMP-2, and MMP-9 in EVT invasion during spiral artery remodeling were evaluated using human EVT from HTR-8/SVneo cell lines. The expression of MMP-2, MMP-9, and mitogen-activated protein kinase (MAPK), and Akt pathways in HTR-8/SVneo cells treated with recom-binant EG-VEGF alongside anti-PROKR1 and/or anti-PROKR2 antibodies was evaluated using quantitative reverse transcription-PCR and western blotting. Wound-healing and cell invasion assays were performed to assess the migration and invasion of these treated cells. Interestingly, 20 nM EG-VEGF activated ERK1/2 sig-naling and upregulated MMP-2 and MMP-9. This effect was suppressed by anti-PROKR2 antibody via ERK1/2 downregulation. Anti-PROKR2 antibody inhibited the migration and invasion of EG-VEGF-stimulated HTR-8/SVneo cells. Elevated concentrations of EG-VEGF enhance EVT invasion in a human trophoblast cell line by upregulating MMP-2 and MMP-9 via PROKR2. These new insights into the regulation of epithelial cell invasion may help in developing therapeutic interventions for placental-related diseases during pregnancy. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 5 2021 Histologic Transformation from Follicular Lymphoma to Diffuse Large B-cell Lymphoma Detected during Colonoscopy 625 629 EN Masaya Iwamuro Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasushi Yamasaki Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takehiro Tanaka Department of Pathology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Noboru Asada Department of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ken-ichi Matsuoka Department of Hematology and Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Sakiko Hiraoka Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yoshiro Kawahara Department of Practical Gastrointestinal Endoscopy, Okayama University Hospital Hiroyuki Okada Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Case Report 10.18926/AMO/62775 A 77-year-old Japanese woman who had been treated for follicular lymphoma for 8 years developed abdominal pain and intra-abdominal lymphadenopathies. Colonoscopy revealed an elevated lesion in the rectum, which presented as two humps with erosions. A diagnosis of histologic transformation of follicular lymphoma to diffuse large B-cell lymphoma was made by endoscopic biopsy. This case underscores the importance of endoscopy examinations and biopsy of newly emerged gastrointestinal lesions for the prompt diagnosis of histologic transformation, since salvage chemotherapy must be initiated quickly in such cases. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 5 2021 The Effect of Peer Instruction Lectures on Learning Attitudes in Epidemiology Education 601 609 EN Toshiharu Mitsuhashi Center for Innovative Clinical Medicine, Okayama University Hospital Original Article 10.18926/AMO/62773 Research suggests that the fundamental concepts of epidemiology cannot be sufficiently learned in traditional lectures, and interactive learning is necessary. However, few studies have investigated interactive epidemiology education in general, or peer instruction (PI) in particular. This study investigated the effect of PI. Study par-ticipants were fourth-year medical students. The attitude of participants in regard to PI learning was examined in a non-PI and a PI group. The Survey of Attitudes Toward Statistics (SATS) (containing six sub-categories) was conducted as a learning-attitudes index. The pre- and post-lecture scores were compared between the non-PI and PI groups using double robust (DR) estimation. The non-PI and PI groups consisted of 20 and 121 student participants, respectively. In DR estimation, affect exhibited the lowest SATS score changes, at −0.51 (95% confidence interval −0.78 to −0.24; p-value < 0.001), whereas effort exhibited the highest score changes of 0.01 (95% confidence interval −0.30 to 0.32; p-value = 0.952). The epidemiology lecture with PI did not increase the SATS scores. This might be due to issues related to the experimental design. Further research investigating the effects of interactive epidemiology education, it will be necessary to develop tools for assessing the learning of epidemiological concepts and to improve the research design. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 5 2021 Glial Cells as Possible Targets of Neuroprotection through Neurotrophic and Antioxidative Molecules in the Central and Enteric Nervous Systems in Parkinson’s Disease 549 556 EN Nami Isooka Department of Medical Neurobiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ikuko Miyazaki Department of Medical Neurobiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masato Asanuma Department of Medical Neurobiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Review 10.18926/AMO/62767 Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide. The loss of nigrostriatal dopaminergic neurons produces its characteristic motor symptoms, but PD patients also have non-motor symptoms such as constipation and orthostatic hypotension. The pathological hallmark of PD is the presence of α-synuclein-containing Lewy bodies and neurites in the brain. However, the PD pathology is observed in not only the central nervous system (CNS) but also in parts of the peripheral nervous system such as the enteric nervous system (ENS). Since constipation is a typical prodromal non-motor symptom in PD, often preceding motor symptoms by 10-20 years, it has been hypothesized that PD pathology propagates from the ENS to the CNS via the vagal nerve. Discovery of pharmacological and other methods to halt this progression of neurodegeneration in PD has the potential to improve millions of lives. Astrocytes protect neurons in the CNS by secretion of neurotrophic and antioxidative factors. Similarly, astrocyte-like enteric glial cells (EGCs) are known to secrete neuroprotective factors in the ENS. In this article, we summarize the neuroprotective function of astrocytes and EGCs and discuss therapeutic strategies for the prevention of neurodegeneration in PD targeting neurotrophic and antioxidative molecules in glial cells. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0026-265X 171 2021 Portable two-color photometer based on paired light emitter detector diodes and its application to the determination of paraquat and diquat 106777 EN Sasikarn Seetasang Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University Takashi Kaneta Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University Here we describe a methodology for the determination of paraquat and diquat using a newly developed portable photometer equipped with two colors of paired light emitter detector diodes (PEDD). The colorimetric measurements employed in this work include the redox reactions between 1) dithiothreitol and diquat to produce the red color characteristic of a diquat radical and 2) between sodium dithionite and either diquat or paraquat that results in the green and blue colors of diquat and paraquat radicals, respectively. The addition of sodium dithionite or dithiothreitol in a solid-state provides reproducible absorbance of the radicals, prevents decomposition of the reagents in a solution, and simplifies handling of the reagents. The diquat radical produced by dithiothreitol (λmax = 495 nm) was successfully detected by using a pair of blue LEDs with a maximum emission wavelength at 472 nm while the radicals of paraquat (λmax = 603 nm) and diquat (λmax = 771 nm) reduced by sodium dithionite were measured by a pair of orange LEDs with a maximum emission wavelength of 609 nm. The proposed method consists of measuring diquat radicals at 472 nm, estimating the absorbance of diquat radicals at 609 nm, and subtracting the estimated absorbance of diquat radicals from the total absorbance at 609 nm to determine paraquat radicals. The developed method yielded examples of excellent linear regression (r2) of more than 0.99 in three calibration curves of the radicals measured at 472 nm for diquat radicals and measured at 609 nm for both diquat and paraquat radicals. The intra-day (n = 3) and inter-day (n = 3) precision of three calibration curves were less than or equal to 5%. By comparison with the standard method of high-performance liquid chromatography, the reliability of the proposed method was proven via the analysis of paraquat and diquat radicals in a commercially available herbicide. No potential conflict of interest relevant to this article was reported.

Elsevier B.V. Acta Medica Okayama 09240136 299 2021 Effects of Superposition of 532 nm and 1064 nm Wavelengths in Copper Micro-welding by Pulsed Nd:YAG Laser 117388 EN Martin Ruthandi Maina Nontraditional Machining Laboratory, Okayama University Yasuhiro Okamoto Nontraditional Machining Laboratory, Okayama University Kazuki Hamada Nontraditional Machining Laboratory, Okayama University Akira Okada Nontraditional Machining Laboratory, Okayama University Shin-ichi Nakashiba Kataoka Corporation Norio Nishi Kataoka Corporation Unstable and low absorption of laser energy is experienced in copper welding at around 1000 nm wavelength. At 532 nm wavelength, there is stable and high laser absorption by copper. Past researches have shown that transitional processing condition between keyhole and heat conduction welding results in a stable micro-welding process characterized by good surface quality and deep penetration. In order to adapt laser welding to copper using pulsed Nd:YAG lasers, investigations of welding quality and efficiency were addressed. Processing was done under transitional processing condition between heat conduction and keyhole welding. Copper C1020 specimens were processed using superposed laser wavelengths of 1064 nm and 532 nm. Effects of irradiation delay and power density on the process were clarified by taking measurements of absorption rates and molten volumes, and by analyzing the weld beads. In addition, the dynamics of molten area and keyhole formation were investigated through three-dimensional FEM analysis. A stabilized laser absorption and increased molten volume was achieved by superposition using 532 nm laser of an appropriate high power density coupled with a short irradiation delay for the 1064 nm laser, which resulted in high-efficiency welding of copper. No potential conflict of interest relevant to this article was reported.

Nature Portfolio Acta Medica Okayama 2045-2322 11 1 2021 Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization 14765 EN Masuzu Kikuchi Division of Pharmaceutical Sciences, Okayama University Keiichi Kojima Division of Pharmaceutical Sciences, Okayama University Shin Nakao Division of Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Shiho Kawanishi Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Atsushi Shibukawa Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takashi Kikukawa Faculty of Advanced Life Science, Hokkaido University Yuki Sudo Division of Pharmaceutical Sciences, Okayama University Microbial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pK(a)=3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 4 2021 Arrhythmogenic Right Ventricular Cardiomyopathy Diagnosed during Hospitalization for Cardiac Arrest 517 521 EN Masahiko Ochi Department of Cardiology, Japan Red Cross Okayama Hospital Atsuyoshi Iida Department of Emergency Medicine, Japan Red Cross Okayama Hospital Yuka Takahashi Division of Pathology and Clinical Laboratories, National Cancer Center Hospital Masamichi Tanaka Department of Cardiology, Japan Red Cross Okayama Hospital Hironori Saito Department of Cardiology, Japan Red Cross Okayama Hospital Hiromichi Naito Department of Emergency, Critical Care, and Disaster Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takeshi Mikane Department of Emergency Medicine, Japan Red Cross Okayama Hospital Soichiro Fuke Department of Cardiology, Japan Red Cross Okayama Hospital Case Report 10.18926/AMO/62405 Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a genetically mediated cardiomyopathy charac-terized by progressive myocardial loss of the right ventricle and its replacement by fibrofatty tissue, causing dyskinesia, aneurysm, and/or arrhythmia. The prevalence of ARVC is estimated to be 1 in 2,000-5,000, with the condition accounting for up to 20% of sudden cardiac deaths in individuals < 35 years old. This report describes the case of 61-year-old Japanese who was diagnosed with ARVC after cardiac arrest (CA) and successful resusci-tation. After the sudden CA, the restoration of spontaneous circulation was achieved with appropriate resusci-tation, followed by the introduction of target temperature management in the intensive care unit. He was diag-nosed with ARVC based on angiography and histology results. An ICD (implantable cardioverter-defibrillator) was implanted, and he was discharged without neurological sequelae 1 month post-CA. ARVC is an important cause of sudden CA, and successfully resuscitated patients with right ventricular dilation should undergo testing to rule out ARVC. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2073-4409 10 6 2021 Exosome-Based Molecular Transfer Activity of Macrophage-Like Cells Involves Viability of Oral Carcinoma Cells: Size Exclusion Chromatography and Concentration Filter Method 1328 EN Yanyin Lu Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takanori Eguchi Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Chiharu Sogawa Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Eman A. Taha Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Manh Tien Tran Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiki Nara Research Program for Undergraduate Students, Okayama University Dental School Penggong Wei Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shiro Fukuoka Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takuya Miyawaki Department of Dental Anesthesiology and Special Care Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kuniaki Okamoto Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Extracellular vesicles (EV) heterogeneity is a crucial issue in biology and medicine. In addition, tumor-associated macrophages are key components in cancer microenvironment and immunology. We developed a combination method of size exclusion chromatography and concentration filters (SEC-CF) and aimed to characterize different EV types by their size, cargo types, and functions. A human monocytic leukemia cell line THP-1 was differentiated to CD14-positive macrophage-like cells by stimulation with PMA (phorbol 12-myristate 13-acetate) but not M1 or M2 types. Using the SEC-CF method, the following five EV types were fractionated from the culture supernatant of macrophage-like cells: (i) rare large EVs (500-3000 nm) reminiscent of apoptosomes, (ii) EVs (100-500 nm) reminiscent of microvesicles (or microparticles), (iii) EVs (80-300 nm) containing CD9-positive large exosomes (EXO-L), (iv) EVs (20-200 nm) containing unidentified vesicles/particles, and (v) EVs (10-70 nm) containing CD63/HSP90-positive small exosomes (EXO-S) and particles. For a molecular transfer assay, we developed a THP-1-based stable cell line producing a GFP-fused palmitoylation signal (palmGFP) associated with the membrane. The THP1/palmGFP cells were differentiated into macrophages producing palmGFP-contained EVs. The macrophage/palmGFP-secreted EXO-S and EXO-L efficiently transferred the palmGFP to receiver human oral carcinoma cells (HSC-3/palmTomato), as compared to other EV types. In addition, the macrophage-secreted EXO-S and EXO-L significantly reduced the cell viability (ATP content) in oral carcinoma cells. Taken together, the SEC-CF method is useful for the purification of large and small exosomes with higher molecular transfer activities, enabling efficient molecular delivery to target cells. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 3 2021 A Case of Metastatic Fumarate Hydratase-Deficient–like Renal Cell Carcinoma Successfully Managed by Ipilimumab plus Nivolumab 397 402 EN Takanori Sekito Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Atsushi Takamoto Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuyuki Kobayashi Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masao Mitsui Department of Urology, Kurashiki Medical Center Shogo Watari Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Risa Kubota Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takuya Sadahira Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takehiro Iwata Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shingo Nishimura Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kohei Edamura Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomoko Sako Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Motoo Araki Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masami Watanabe Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toyohiko Watanabe Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Rei Shibata Department of Pathology, Okayama University Hospital Daisuke Ennishi Department of Hematology and Oncology, Okayama University Hospital Yasutomo Nasu Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Case Report 10.18926/AMO/62237 We report a 62-year-old male with metastatic fumarate hydratase-deficient renal cell carcinoma (FH-deficient RCC) without fumarate hydratase (FH) mutation (FH-deficient–like RCC). The International Metastatic RCC Database Consortium risk score was intermediate, and immunotherapy with nivolumab and ipilimumab (Ipi/ Nivo) was initiated. Four cycles of Ipi/Nivo and 5 cycles of nivolumab resulted in a complete response of the metastases. Hypophysitis occurred as an immune-related adverse event after four cycles of Ipi/Nivo. The prognosis of patients with FH-deficient RCC is generally poor. Few reports of FH-deficient RCC successfully treated with Ipi/Nivo have been published. Ipi/Nivo can be effective for treating FH-deficient RCC. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 3 2021 Texture Indices of 18F-FDG PET/CT for Differentiating Squamous Cell Carcinoma and Non-Hodgkin’s Lymphoma of the Oropharynx 351 356 EN Katsuya Mitamura Department of Radiology, Faculty of Medicine, Kagawa University Takashi Norikane Department of Radiology, Faculty of Medicine, Kagawa University Yuka Yamamoto Department of Radiology, Faculty of Medicine, Kagawa University Ayumi Ihara-Nishishita Department of Radiology, Faculty of Medicine, Kagawa University Takuya Kobata Department of Radiology, Faculty of Medicine, Kagawa University Kengo Fujimoto Department of Radiology, Faculty of Medicine, Kagawa University Yasukage Takami Department of Radiology, Faculty of Medicine, Kagawa University Nobuyuki Kudomi Department of Medical Physics, Faculty of Medicine, Kagawa University Hiroshi Hoshikawa Department of Otolaryngology, Faculty of Medicine, Kagawa University Yoshihiro Nishiyama Department of Radiology, Faculty of Medicine, Kagawa University Original Article 10.18926/AMO/62230 We assessed the role of 18F-FDG PET/CT texture indices for the differentiation of squamous cell carcinoma (SCC) and non-Hodgkin’s lymphoma (NHL) in the oropharynx. 18F-FDG PET/CT data for 27 patients with SCC and 25 patients with NHL in the oropharynx were investigated. The maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV), total lesion glycolysis (TLG), and six texture indices (homogeneity, entropy, short-run emphasis, long-run emphasis, low gray-level zone emphasis [LGZE], and high graylevel zone emphasis [HGZE]) were derived from PET images. PET/CT parameters of the SCC patients were compared with those of the NHL patients. The diagnostic accuracy of the indices for differentiating SCC from NHL was calculated by a receiver operating characteristic curve analysis. 18F-FDG uptake in the oropharynx was observed in all of the patients. The SUVmax, MTV, and TLG did not differ significantly between the SCC and NHL groups, but two of the six texture indices (LGZE [p=0.004] and HGZE [p=0.03]) showed significant differences between the groups. LGZE was the best discriminative index for the differentiation of SCC and NHL (55.6% sensitivity, 88.0% specificity). The LGZE and HGZE texture indices derived from 18F-FDG PET/CT images may be useful in differentiating SCC and NHL in the oropharynx. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 3 2021 Dental Treatment for Special Needs Patients Under General Anaesthesia: A 14-year Experience from South Bosnia and Herzegovina 261 268 EN Lidija Lasic Arapovic Health Care Center Mostar, University of Mostar Zoran Karlovic School of Medicine, University of Mostar Valentina Rajic Brzovic School of Medicine, University of Mostar Amer Bukvic Primary Health Care Center Novi Travnik Anka Coric Health Care Center Mostar, University of Mostar Katarina Vukojevic School of Medicine, University of Mostar Zeljko Verzak School of Medicine, University of Mostar Original Article 10.18926/AMO/62217 We conducted a retrospective analysis of records of special needs patients (SNPs) who received dental treatment under orotracheal-intubation general anaesthesia (OIGA) at Caritas Centre St. Family in Mostar, Bosnia and Herzegovina during the 14-year period from January 2005 to December 2018. Of the 7,085 SNPs who received dental treatment, 1,220 (17.2%) received dental treatment under OIGA: 829 (67.9%) males and 391 (32.1%) females. The patients’ mean age was 18.3±10.9 years (747 paediatric and 473 adult patients). Mental retardation and psychiatric problems were the most common medical conditions (81.22%). The most common indication for dental treatment under OIGA was behaviour management (87.21%), and 81% of the patients had an urgent need for treatment. Many of the patients had restorative treatment (3,833) and tooth extractions (3,681). From 2011 onwards, the number of tooth extractions decreased significantly. Annual trends revealed a rapid increase of patients every year. The mean dental treatment duration was 95.3±12.1 min; the mean time under OIGA was 98±8.5 min. No serious adverse effects occurred. There was increase of annual trend of SNP in OIGA. The number of extractions decreased while the number of preventive and restorative dental treatments increased. No potential conflict of interest relevant to this article was reported.

Public Library Science Acta Medica Okayama 1932-6203 16 4 2021 Oncolytic virotherapy promotes radiosensitivity in soft tissue sarcoma by suppressing anti-apoptotic MCL1 expression e0250643 EN Toshinori Omori Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroshi Tazawa Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuaki Yamakawa Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shuhei Osaki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Joe Hasei Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kazuhisa Sugiu Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tadashi Komatsubara Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomohiro Fujiwara Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Aki Yoshida Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiyuki Kunisada Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuo Urata Oncolys BioPharma, Inc. Shunsuke Kagawa Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshifumi Ozaki Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiyoshi Fujiwara Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Soft tissue sarcoma (STS) is a rare cancer that develops from soft tissues in any part of the body. Despite major advances in the treatment of STS, patients are often refractory to conventional radiotherapy, leading to poor prognosis. Enhancement of sensitivity to radiotherapy would therefore improve the clinical outcome of STS patients. We previously revealed that the tumor-specific, replication-competent oncolytic adenovirus OBP-301 kills human sarcoma cells. In this study, we investigated the radiosensitizing effect of OBP-301 in human STS cells. The in vitro antitumor effect of OBP-301 and ionizing radiation in monotherapy or combination therapy was assessed using highly radiosensitive (RD-ES and SK-ES-1) and moderately radiosensitive (HT1080 and NMS-2) STS cell lines. The expression of markers for apoptosis and DNA damage were evaluated in STS cells after treatment. The therapeutic potential of combination therapy was further analyzed using SK-ES-1 and HT1080 cells in subcutaneous xenograft tumor models. The combination of OBP-301 and ionizing radiation showed a synergistic antitumor effect in all human STS cell lines tested, including those that show different radiosensitivity. OBP-301 was found to enhance irradiation-induced apoptosis and DNA damage via suppression of anti-apoptotic myeloid cell leukemia 1 (MCL1), which was expressed at higher levels in moderately radiosensitive cell lines. The combination of OBP-301 and ionizing radiation showed a more profound antitumor effect compared to monotherapy in SK-ES-1 (highly radiosensitive) and HT1080 (moderately radiosensitive) subcutaneous xenograft tumors. OBP-301 is a promising antitumor reagent to improve the therapeutic potential of radiotherapy by increasing radiation-induced apoptosis in STS. No potential conflict of interest relevant to this article was reported.

Dove Medical Press Ltd. Acta Medica Okayama 1178-2013 16 2021 Dual-Functional PLGA Nanoparticles Co-Loaded with Indocyanine Green and Resiquimod for Prostate Cancer Treatment 2775 2787 EN Wenfeng Lin Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Chaoming Li Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Naijin Xu Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masami Watanabe Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ruizhi Xue Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Abai Xu Department of Urology, Zhujiang Hospital, Southern Medical University Motoo Araki Okayama University Hospital Ruifen Sun Center for Scientific Research, Yunnan University of Chinese Traditional Medicine Chunxiao Liu Department of Urology, Zhujiang Hospital, Southern Medical University Yasutomo Nasu Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Peng Huang Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Purpose: With the advance of screening techniques, there is a growing number of low-risk or intermediate-risk prostate cancer (PCa) cases, remaining a serious threat to men's health. To obtain better efficacy, a growing interest has been attracted to develop such emerging treatments as immunotherapy and focal therapy. However, few studies offer guidance on whether and how to combine these modalities against PCa. This study was designed to develop dual-functional nanoparticles (NPs) which combined photothermal therapy (PTT) with immunotherapy and determine the anti-tumor efficacy for PCa treatment. Methods: By a double emulsion technique, the drug nanocarrier, poly(lactic-co-glycolic acid) or PLGA, was applied for co-loading of a fluorescent dye, indocyanine green (ICG) and a toll-like receptor 7/8 (TLR7/8) agonist resiquimod (R848) to synthesize PLGA-ICG-R848 NPs. Next, we determined their characteristic features and evaluated whether they inhibited the cell viability in multiple PCa cell lines. After treatment with PLGA-ICG-R848, the maturation markers of bone marrow-derived dendritic cells (BMDCs) were detected by flow cytometry. By establishing a subcutaneous xenograft model of mouse PCa, we explored both the anti-tumor effect and immune response following the NPs-based laser ablation. Results: With a mean diameter of 157.7 nm, PLGA-ICG-R848 exhibited no cytotoxic effect in PCa cells, but they significantly decreased RM9 cell viability to (3.9 +/- 1.0)% after laser irradiation. Moreover, PLGA-ICG-R848 promoted BMDCs maturation with the significantly elevated proportions of CD11c+CD86+ and CD11c+CD80+ cells. Following PLGA-ICG-R848-based laser ablation in vivo, the decreased bioluminescent signals indicated a significant inhibition of PCa growth, while the ratio of splenic natural killer (NK) cells in PLGA-ICG-R848 was (3.96 +/- 1.88)% compared with (0.99 +/- 0.10)% in PBS group, revealing the enhanced immune response against PCa. Conclusion: The dual-functional PLGA-ICG-R848 NPs under laser irradiation exhibit the anti-tumor efficacy for PCa treatment by combining PTT with immunotherapy. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 2 2021 Pyogenic Ventriculitis After Anterior Skull Base Surgery Treated With Endoscopic Ventricular Irrigation And Reconstruction Using a Vascularized Flap 243 248 EN Yusuke Tomita Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yosuke Shimazu Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Masato Kawakami Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Hiroshi Matsumoto Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Kentaro Fujii Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Masahiro Kameda Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Takao Yasuhara Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yasuki Suruga Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Tomoyuki Ota Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Yoshihiro Kimata Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Kazuhiko Kurozumi Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Isao Date Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Case Report 10.18926/AMO/61908 Ventriculitis is a rare, serious complication of neurosurgery. A 59-year-old man who had undergone a craniotomy for a paranasal adenocarcinoma, developed a right frontal cystic lesion. We performed a bifrontal craniotomy to remove the lesion. The dura was repaired with non-vascularized free fascia lata in watertight fashion. Ventriculitis occurred 3 days postoperatively. Ventricular drainage, craniectomy, and endoscopic irrigation were undertaken to remove an abscess. The dura and the resection cavity were reconstructed using a vascularized anterolateral thigh adipofascial flap. His symptoms disappeared, indicating that endoscopic irrigation and reconstruction can effectively address ventriculitis even in patients in critical clinical condition. No potential conflict of interest relevant to this article was reported.

AIP Publishing Acta Medica Okayama 0021-9606 154 9 2021 Formation of hot ice caused by carbon nanobrushes. II. Dependency on the radius of nanotubes 094502 EN Masakazu Matsumoto Research Institute for Interdisciplinary Science, Okayama University Takuma Yagasaki Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University Hideki Tanaka Research Institute for Interdisciplinary Science, Okayama University Stable crystalline structures of confined water can be different from bulk ice. In Paper I [T. Yagasaki et al., J. Chem. Phys. 151, 064702 (2019)] of this study, it was shown, using molecular dynamics (MD) simulations, that a zeolite-like ice structure forms in nanobrushes consisting of (6,6) carbon nanotubes (CNTs) when the CNTs are located in a triangle arrangement. The melting temperature of the zeolite-like ice structure is much higher than the melting temperature of ice Ih when the distance between the surfaces of CNTs is ∼0.94 nm, which is the best spacing for the bilayer structure of water. In this paper, we perform MD simulations of nanobrushes of CNTs that are different from (6,6) CNTs in radius. Several new porous ice structures form spontaneously in the MD simulations. A stable porous ice forms when the radius of its cavities matches the radius of the CNTs well. All cylindrical porous ice structures found in this study can be decomposed into a small number of structural blocks. We provide a new protocol to classify cylindrical porous ice crystals on the basis of this decomposition. No potential conflict of interest relevant to this article was reported.

IOP Publishing Acta Medica Okayama 1367-2630 21 2019 Superconductivity in a new layered triangular-lattice system Li2IrSi2 093056 EN K Horigane Research Institute for Interdisciplinary Science, Okayama University, K Takeuchi Graduate School of natural science and technology, Okayama University D Hyakumura Department of Physics and Mathematics, Aoyama Gakuin University R Horie Research Institute for Interdisciplinary Science, Okayama University T Sato Institute for Materials Research, Tohoku University T Muranaka Department of Engineering Science, University of Electro-Communications K Kawashima Department of Physics and Mathematics, Aoyama Gakuin University H Ishii National Synchrotron Radiation Research Center Y Kubozono Research Institute for Interdisciplinary Science, Okayama University S Orimo Institute for Materials Research, Tohoku University M Isobe National Institute for Materials Science (NIMS) J Akimitsu Research Institute for Interdisciplinary Science, Okayama University We report on the crystal structure and superconducting properties of a novel iridium-silicide, namely Li2IrSi2. It has a Ag2NiO2-type structure (space group R-3m) with the lattice parameters a = 4.028 30(6) Å and c = 13.161 80(15) Å. The crystal structure comprises IrSi2 and double Li layers stacked alternately along the c-axis. The IrSi2 layer includes a two-dimensional Ir equilateral-triangular lattice. Electrical resistivity and static magnetic measurements revealed that Li2IrSi2 is a type-II superconductor with critical temperature (Tc) of 3.3 K. We estimated the following superconducting parameters: lower critical field Hc1(0) ~ 42 Oe, upper critical field Hc2(0) ~ 1.7 kOe, penetration depth λ0 ~ 265 nm, coherence length ξ0 ~ 44 nm, and Ginzburg–Landau parameter κGL ~ 6.02. The specific-heat data suggested that superconductivity in Li2IrSi2 could be attributed to weak-coupling Cooper pairs. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 22 4 2021 A Novel Artificially Humanized Anti-Cripto-1 Antibody Suppressing Cancer Cell Growth 1709 EN Hiroko Ishii GSP Enterprise, Inc. Maram H. Zahra Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Atushi Takayanagi GSP Enterprise, Inc. Masaharu Seno Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Cripto-1 is a member of the EGF-CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti-Cripto-1 artificial antibody and assessed the recognition to the antigen and the potential to suppress the growth of cancer stem cells. First, single chain antibody clones were isolated by bio-panning with the affinity to recombinant Cripto-1 protein from our original phage-display library. Then, the variable regions of heavy chain VH and light chain VL in each clone were fused to constant regions of heavy chain CH and light chain CL regions respectively. These fused genes were expressed in ExpiCHO-S cells to produce artificial humanized antibodies against Cripto-1. After evaluation of the expression levels, one clone was selected and the anti-Cripto-1 antibody was produced and purified. The purified antibody showed affinity to recombinant Cripto-1 at 1.1 pmol and immunoreactivity to cancer tissues and cell lines. The antibody was available to detect the immunoreactivity in tissue microarrays of malignant tumors as well as in Cripto-1 overexpressing cells. Simultaneously, the antibody exhibited the potential to suppress the growth of human colon cancer derived GEO cells overexpressing Cripto-1 with IC50 at approximately 110 nM. The artificially humanized antibody is proposed to be a good candidate to target cancer cells overexpressing Cripto-1. No potential conflict of interest relevant to this article was reported.

Nature Research Acta Medica Okayama 2045-2322 10 1 2020 Lokiarchaeota archaeon schizorhodopsin-2 (LaSzR2) is an inward proton pump displaying a characteristic feature of acid-induced spectral blue-shift 20857 EN Keiichi Kojima Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Masumi Hasegawa Atmosphere and Ocean Research Institute, The University of Tokyo Masaki Nakama Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Marie Kurihara Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takashi Kikukawa Faculty of Advanced Life Science, Hokkaido University Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University The photoreactive protein rhodopsin is widespread in microorganisms and has a variety of photobiological functions. Recently, a novel phylogenetically distinctive group named 'schizorhodopsin (SzR)' has been identified as an inward proton pump. We performed functional and spectroscopic studies on an uncharacterised schizorhodopsin from the phylum Lokiarchaeota archaeon. The protein, LaSzR2, having an all-trans-retinal chromophore, showed inward proton pump activity with an absorption maximum at 549 nm. The pH titration experiments revealed that the protonated Schiff base of the retinal chromophore (Lys188, pK(a)=12.3) is stabilised by the deprotonated counterion (presumably Asp184, pK(a)=3.7). The flash-photolysis experiments revealed the presence of two photointermediates, K and M. A proton was released and uptaken from bulk solution upon the formation and decay of the M intermediate. During the M-decay, the Schiff base was reprotonated by the proton from a proton donating residue (presumably Asp172). These properties were compared with other inward (SzRs and xenorhodopsins, XeRs) and outward proton pumps. Notably, LaSzR2 showed acid-induced spectral 'blue-shift' due to the protonation of the counterion, whereas outward proton pumps showed opposite shifts (red-shifts). Thus, we can distinguish between inward and outward proton pumps by the direction of the acid-induced spectral shift. No potential conflict of interest relevant to this article was reported.

Springer Acta Medica Okayama 0168-8162 30 1-3 2003 Orchid Fleck Virus: Brevipalpus californicus Mite Transmission, Biological Properties and Genome Structure 215 223 EN Hideki Kondo Research Institute for Bioresources, Okayama University Takanori Maeda Research Institute for Bioresources, Okayama University Tetsuo Tamada Research Institute for Bioresources, Okayama University Orchid fleck virus (OFV) causes necrotic or chlorotic ring spots and fleck symptoms in many orchid species world-wide. The virus has non-enveloped, bacilliform particles of about 40 nm × 100–150 nm and is sap-transmissible to several plant species. OFV is transmitted by the mite Brevipalpus californicus (Banks) in a persistent manner and efficiently transmitted by both adults and nymphs, but not by larvae. Viruliferous mites retain their infectivity for 3 weeks on a virus-immune host. The genome of OFV consists of two molecules of 6431 (RNA1) and 6001 nucleotides (RNA2). The RNAs have conserved and complementary terminal sequences. RNA1 contains five open reading frames (ORF), and RNA2 encodes a single ORF. Although some of the encoded proteins of OFV have sequences similar to those of proteins of plant rhabdoviruses, OFV differs from viruses in the family Rhabdoviridae in having a bipartite genome. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 5 2020 Combined Laparoscopic and CT Monitoring of the Ice-Ball Margin during Cryoablation for Renal Cell Carcinoma Associated with von Hippel-Lindau Disease: First Case 443 448 EN Takanori Sekito Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Motoo Araki Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takao Hiraki Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mayu Uka Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiyuki Komaki Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yusuke Matsui Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshihiro Iguchi Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Satoshi Katayama Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kasumi Yoshinaga Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shogo Watari Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yuki Maruyama Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yosuke Mitsui Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Risa Kubota Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takuya Sadahira Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shingo Nishimura Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Koichiro Wada Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Atsushi Takamoto Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kohei Edamura Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomoko Sako Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuyuki Kobayashi Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toyohiko Watanabe Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Susumu Kanazawa Department of Radiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasutomo Nasu Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Case Report 10.18926/AMO/60806 We report a 47-year-old Japanese female with 10 previous treatments for multiple bilateral renal cell carcinoma (RCC) associated with von Hippel-Lindau disease. The 14-mm right lower pole renal tumor was in contact with the right ureter. Laparoscopic cryoablation was performed to protect the ureter wrapped with gauze. Computed tomography (CT) monitoring was used to confirm the precise ≥ 6 mm ice-ball margin. There was no local progression at 6-months post-surgery. The serum creatinine has been stable. This is apparently the first report of combined laparoscopic and CT monitoring of an ice-ball formation and its margin during cryoablation for RCC. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 5 2020 Oblique Surface Dose Calculation in High-Energy X-ray Therapy 415 422 EN Naomasa Narihiro Graduate School of Health Sciences, Department of Radiological Technology, Okayama University Masataka Oita Department of Graduate School of Interdisciplinary Sciences and Engineering in Health Systems, Okayama University Yoshihiro Takeda Graduate School of Health Sciences, Department of Radiological Technology, Okayama University Original Article 10.18926/AMO/60801 During radiation therapy, incident radiation oblique to the skin surface is high and may cause severe skin damage. Understanding the dose of radiation absorbed by the skin is important for predicting skin damage due to radiation. In this study, we used a high-energy (4 MV) X-ray system and an optically stimulated luminescence dosimeter (OSLD) that was developed for personal exposure dosimetry. We determined the dose variation and angular dependence, which are the characteristics of a small OSLD required to derive the calculation formula for the oblique surface dose. The dose variation was determined using the coefficient of variation. The maximum coefficient of variation for 66 small-field OSLDs was 1.71%. The angular dependence, obtained from the dose ratio of the dosimeter in the vertical direction, had a maximum value of 1.37. We derived a new equation in which the oblique surface dose can be calculated within the error range of −7.7-5.1%. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 5 2020 Evaluation of Prognosis of Juvenile Differentiated Thyroid Carcinoma 401 406 EN Takahiro Tsukioki Department of Breast and Endocrine Surgery, Okayama University Hospital Tadahiko Shien Department of Breast and Endocrine Surgery, Okayama University Hospital Yusuke Ohtani Department of Breast and Endocrine Surgery, Okayama University Hospital Miwa Fujihara Department of Breast and Endocrine Surgery, Okayama University Hospital Yoko Suzuki Department of Breast and Endocrine Surgery, Okayama University Hospital Yukiko Kajihara Department of Breast and Endocrine Surgery, Okayama University Hospital Minami Hatono Department of Breast and Endocrine Surgery, Okayama University Hospital Kengo Kawada Department of Breast and Endocrine Surgery, Okayama University Hospital Mariko Kochi Department of Breast and Endocrine Surgery, Okayama University Hospital Takayuki Iwamoto Department of Breast and Endocrine Surgery, Okayama University Hospital Hirokuni Ikeda Department of Breast and Endocrine Surgery, Okayama University Hospital Naruto Taira Department of Breast and Endocrine Surgery, Okayama University Hospital Hiroyoshi Doihara Department of Breast and Endocrine Surgery, Okayama University Hospital Original Article 10.18926/AMO/60799 Differentiated thyroid carcinoma (DTC) in juvenile patients is often an extensive and aggressive disease with a high frequency of recurrence. However, the prognosis is excellent, with a low mortality rate even when advanced disease is present, although prognostic factors and treatment strategy remain uncertain. Between April 2004 and March 2017, 33 juvenile patients (< 30 years old) were diagnosed with DTC and treated at our institution. We retrospectively investigated prognosis and factors including sex, reason for discovery, treatment, pathological factors and treatment progress to clarify the risk factors. All patients underwent curative surgical treatment. Pathologically, lymph node metastasis was identified in 25 patients (75%). Thirteen patients (39%) had bilateral cervical metastasis. In addition, 9 (27%) had more than 10 metastatic lymph nodes. The 2 patients with more than 20 metastatic lymph nodes were treated with radioactive iodine (RAI). Five patients (15%) had local recurrences and received surgery. There have been no further recurrences or deaths. However, no factors were determined to significantly predict the recurrence of juvenile DTC. Local recurrent disease was treated with surgery and/or RAI until remission, and survival was excellent in juvenile DTC. No potential conflict of interest relevant to this article was reported.

Elsevier Acta Medica Okayama 0005-2728 1861 7 2020 Spectral tuning of light-harvesting complex II in the siphonous alga Bryopsis corticulans and its effect on energy transfer dynamics 148191 EN Parveen Akhtar Biological Research Centre Paweł J. Nowakowski ivision of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University Wenda Wang Photosynthesis Research Centre, Chinese Academy of Sciences Thanh Nhut Do Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University Songhao Zhao Photosynthesis Research Centre, Chinese Academy of Sciences Giuliano Siligardi Diamond Light Source Ltd., Harwell Science and Innovation Campus Győző Garab Biological Research Centre Jian-Ren Shen Research Institute for Interdisciplinary Science, Graduate School of Natural Science and Technology, Okayama University Howe-Siang Tan Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University Petar H. Lambrev Biological Research Centre Light-harvesting complex II (LHCII) from the marine green macroalga Bryopsis corticulans is spectroscopically characterized to understand the structural and functional changes resulting from adaptation to intertidal environment. LHCII is homologous to its counterpart in land plants but has a different carotenoid and chlorophyll (Chl) composition. This is reflected in the steady-state absorption, fluorescence, linear dichroism, circular dichroism and anisotropic circular dichroism spectra. Time-resolved fluorescence and two-dimensional electronic spectroscopy were used to investigate the consequences of this adaptive change in the pigment composition on the excited-state dynamics. The complex contains additional Chl b spectral forms – absorbing at around 650 nm and 658 nm – and lacks the red-most Chl a forms compared with higher-plant LHCII. Similar to plant LHCII, energy transfer between Chls occurs on timescales from under hundred fs (mainly from Chl b to Chl a) to several picoseconds (mainly between Chl a pools). However, the presence of long-lived, weakly coupled Chl b and Chl a states leads to slower exciton equilibration in LHCII from B. corticulans. The finding demonstrates a trade-off between the enhanced absorption of blue-green light and the excitation migration time. However, the adaptive change does not result in a significant drop in the overall photochemical efficiency of Photosystem II. These results show that LHCII is a robust adaptable system whose spectral properties can be tuned to the environment for optimal light harvesting. No potential conflict of interest relevant to this article was reported.

Elsevier Acta Medica Okayama 00032670 1135 2020 On-site analysis of paraquat using a completely portable photometric detector operated with small, rechargeable batteries 99 106 EN Sasikarn Seetasang Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University Takashi Kaneta Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University This work describes a methodology that can be used to achieve on-site analysis of paraquat in water samples by using a miniaturized portable photometer consisting of a couple of light-emitting diodes (LEDs). Paraquat produces a colored radical via a redox reaction with sodium dithionite, which is unstable against oxygen in solution. The steps taken to stabilize the reagent solution included control of the pH and the addition of organic solvents, but the most effective was the formation of an oil layer. Together, these steps stabilized the reagent solution for two days. An increase in the duration of reagent stability, however, is necessary in order to transport the reagent for on-site applications in remote locales. For the time being, an excess amount of solid sodium dithionite can be added directly to sample solutions because the unreacted dithionite shows no influence on absorbance of the paraquat radical. Orange LEDs with a maximum emission wavelength of 609 nm were employed in the portable photometer to measure the absorbance of paraquat radical produced by a redox reaction that has an absorption maximum of 603 nm. The developed photometer showed excellent performance with a linear range of from 2.0 mg L−1 to 40.0 mg L−1 and a linear regression (r2 = 1). The limits of detection and quantification were 0.5 mg L−1 and 1.5 mg L−1, respectively, intra-day precision (n = 3) and inter-day precision (n = 5) were both less than 5%, and accuracy based on the percentage of sample recovery ranged from 89 ± 0 to 105 ± 0% (n = 3). The proposed method was applied to the analysis of paraquat in water samples taken from rice fields. The results showed no paraquat in all thirteen samples, which could have been due to strong adsorption of paraquat by soil particles and/or to complications with the sampling conditions. To confirm the adsorption onto soil of paraquat contained in water, we constructed an artificial rice field where water containing paraquat was impounded above the soil layer. The results showed that paraquat in water gradually decreased within three days and could be measured in the soil on the fourth day. These results were confirmed by HPLC analysis, which underscores the utility of this portable photometer for the on-site monitoring of paraquat in water samples. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 21 15 2020 Cellular Fragments as Biomaterial for Rapid In Vitro Bone-Like Tissue Synthesis 5327 EN Mst Nahid Akhter Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Emilio Satoshi Hara Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Koichi Kadoya Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masahiro Okada Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takuya Matsumoto Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Current stem cell-based techniques for bone-like tissue synthesis require at least two to three weeks. Therefore, novel techniques to promote rapid 3D bone-like tissue synthesis in vitro are still required. In this study, we explored the concept of using cell nanofragments as a substrate material to promote rapid bone formation in vitro. The methods for cell nanofragment fabrication were ultrasonication (30 s and 3 min), non-ionic detergent (triton 0.1% and 1%), or freeze-dried powder. The results showed that ultrasonication for 3 min allowed the fabrication of homogeneous nanofragments of less than 150 nm in length, which mineralized surprisingly in just one day, faster than the fragments obtained from all other methods. Further optimization of culture conditions indicated that a concentration of 10 mM or 100 mM of beta-glycerophosphate enhanced, whereas fetal bovine serum (FBS) inhibited in a concentration-dependent manner, the mineralization of the cell nanofragments. Finally, a 3D collagen-cell nanofragment-mineral complex mimicking a bone-like structure was generated in just two days by combining the cell nanofragments in collagen gel. In conclusion, sonication for three min could be applied as a novel method to fabricate cell nanofragments of less than 150 nm in length, which can be used as a material for in vitro bone tissue engineering. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2076-3417 10 14 2020 Finite Element Study of the Effect of Internal Cracks on Surface Profile Change due to Low Loading of Turbine Blade 4883 EN Junji Sakamoto Graduate School of Natural Science and Technology, Okayama University Naoya Tada Graduate School of Natural Science and Technology, Okayama University Takeshi Uemori Graduate School of Natural Science and Technology, Okayama University Hayato Kuniyasu Shimadzu Corporation Turbine blades for thermal power plants are exposed to severe environments, making it necessary to ensure safety against damage, such as crack formation. A previous method detected internal cracks by applying a small load to a target member. Changes in the surface properties of the material were detected before and after the load using a digital holographic microscope and a digital height correlation method. In this study, this technique was applied in combination with finite element analysis using a 2D and 3D model simulating the turbine blades. Analysis clarified that the change in the surface properties under a small load varied according to the presence or absence of a crack, and elucidated the strain distribution that caused the difference in the change. In addition, analyses of the 2D model considering the material anisotropy and thermal barrier coating were conducted. The difference in the change in the surface properties and strain distribution according to the presence or absence of cracks was elucidated. The difference in the change in the top surface height distribution of the materials with and without a crack was directly proportional to the crack length. As the value was large with respect to the vertical resolution of 0.2 nm of the digital holographic microscope, the change could be detected by the microscope. No potential conflict of interest relevant to this article was reported.

Elsevier Acta Medica Okayama 0927-7757 520 2017 Indocyanine green-laden poly(ethylene glycol)-block-polylactide (PEG-b-PLA) nanocapsules incorporating reverse micelles: Effects of PEG-b-PLA composition on the nanocapsule diameter and encapsulation efficiency 764 770 EN Takaichi Watanabe Department of Applied Chemistry and Biotechnology, Graduate School of Natural Science and Technology, Okayama University Yui Sakamoto Department of Applied Chemistry and Biotechnology, Graduate School of Natural Science and Technology, Okayama University Tetsuya Inooka Division of Material and Energy Science, Graduate School of Environmental and Life Science, Okayama University Yukitaka Kimura Division of Material and Energy Science, Graduate School of Environmental and Life Science, Okayama University Tsutomu Ono Department of Applied Chemistry and Biotechnology, Graduate School of Natural Science and Technology, Okayama University Reverse micelles are thermodynamically stable systems, with a capacity to encapsulate hydrophilic molecules in their nanosized core, which is smaller than the core generally obtained with water-in-oil-emulsion droplets. Herein, we present a simple technique for the preparation of poly(ethylene glycol)-block-polylactide (PEG-b-PLA) nanocapsules encapsulating a hydrophilic photosensitizer (indocyanine green, ICG), which exploits reverse micelle formation and subsequent emulsion-solvent diffusion. We establish the effect of the PEG-b-PLA composition and the co-surfactant volume on the diameter and water content of the reverse micelles. We demonstrate that the composition of PEG-b-PLA affects also the diameter and encapsulation efficiency of the resulting nanocapsules. We show that the ICG-laden nanocapsules fabricated under the most optimal conditions have a diameter of approximately 100 nm and an ICG encapsulation efficiency of 58%. We believe that the method proposed here is a promising step towards the preparation of hydrophilic drug-laden polymer nanocapsules with a small diameter and therefore suitable for use in drug delivery applications based on enhanced permeability and retention (EPR) effect-driven passive targeting. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2072-6694 12 6 2020 Paclitaxel and Sorafenib: The Effective Combination of Suppressing the Self-Renewal of Cancer Stem Cells 1360 EN Hend M. Nawara Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Said M. Afify Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Ghmkin Hassan Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Maram H. Zahra Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Marwa N. Atallah Vertebrates Embryology and Comparative Anatomy, Zoology Department, Faculty of Science, Menoufia University Hager Mansour Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hagar A. Abu Quora Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Md Jahangir Alam Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Amira Osman Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Hiroki Kakuta Division of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroki Hamada Department of Life Science, Faculty of Science, Okayama University of Science Akimasa Seno Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Masaharu Seno Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Combination therapy, which is a treatment modality combining two or more therapeutic agents, is considered a cornerstone of cancer therapy. The combination of anticancer drugs, of which functions are different from the other, enhances the efficiency compared to the monotherapy because it targets cancer cells in a synergistic or an additive manner. In this study, the combination of paclitaxel and sorafenib in low concentration was evaluated to target cancer stem cells, miPS-BT549cmP and miPS-Huh7cmP cells, developed from mouse induced pluripotent stem cells. The synergistic effect of paclitaxel and sorafenib on cancer stem cells was assessed by the inhibition of proliferation, self-renewal, colony formation, and differentiation. While the IC(50)values of paclitaxel and sorafenib were approximately ranging between 250 and 300 nM and between 6.5 and 8 mu M, respectively, IC(50)of paclitaxel reduced to 20 and 25 nM, which was not toxic in a single dose, in the presence of 1 mu M sorafenib, which was not toxic to the cells. Then, the synergistic effect was further assessed for the potential of self-renewal of cancer stem cells by sphere formation ability. As a result, 1 mu M of sorafenib significantly enhanced the effect of paclitaxel to suppress the number of spheres. Simultaneously, paclitaxel ranging in 1 to 4 nM significantly suppressed not only the colony formation but also the tube formation of the cancer stem cells in the presence of 1 mu M sorafenib. These results suggest the combination therapy of paclitaxel and sorafenib in low doses should be an attractive approach to target cancer stem cells with fewer side effects. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2072-6694 12 5 2020 Knockout of MMP3 Weakens Solid Tumor Organoids and Cancer Extracellular Vesicles 1260 EN Eman A. Taha Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Chiharu Sogawa Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yuka Okusha Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hotaka Kawai Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences May Wathone Oo Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Abdellatif Elseoudi Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yanyin Lu Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hitoshi Nagatsuka Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Satoshi Kubota Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ayano Satoh Department of Medical Bioengineering, Okayama University Graduate School of Natural Science and Technology Kuniaki Okamoto Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takanori Eguchi Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (200-1000 nm) and small EVs (50-200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 led to a significant reduction in tumoroid size and the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. Moreover, CD63, another member of the tetraspanin family, was significantly reduced only in the EVs fractions of the MMP3-KO cells compared to their counterpart. These weakened phenotypes of MMP3-KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3, resulting in increasing the proliferation and tumorigenesis, indicating crucial roles of MMP3 in tumor progression. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2073-4409 9 3 2020 Cell Stress Induced Stressome Release Including Damaged Membrane Vesicles and Extracellular HSP90 by Prostate Cancer Cells 755 EN Takanori Eguchi Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Chiharu Sogawa Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Kisho Ono Department of Oral and Maxillofacial Surgery, Okayama University Hospital Masaki Matsumoto Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University Tran Manh Tien Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuka Okusha Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Benjamin J. Lang Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School Kuniaki Okamoto Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Stuart K. Calderwood Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School Tumor cells exhibit therapeutic stress resistance-associated secretory phenotype involving extracellular vesicles (EVs) such as oncosomes and heat shock proteins (HSPs). Such a secretory phenotype occurs in response to cell stress and cancer therapeutics. HSPs are stress-responsive molecular chaperones promoting proper protein folding, while also being released from cells with EVs as well as a soluble form known as alarmins. We have here investigated the secretory phenotype of castration-resistant prostate cancer (CRPC) cells using proteome analysis. We have also examined the roles of the key co-chaperone CDC37 in the release of EV proteins including CD9 and epithelial-to-mesenchymal transition (EMT), a key event in tumor progression. EVs derived from CRPC cells promoted EMT in normal prostate epithelial cells. Some HSP family members and their potential receptor CD91/LRP1 were enriched at high levels in CRPC cell-derived EVs among over 700 other protein types found by mass spectrometry. The small EVs (30-200 nm in size) were released even in a non-heated condition from the prostate cancer cells, whereas the EMT-coupled release of EVs (200-500 nm) and damaged membrane vesicles with associated HSP90 alpha was increased after heat shock stress (HSS). GAPDH and lactate dehydrogenase, a marker of membrane leakage/damage, were also found in conditioned media upon HSS. During this stress response, the intracellular chaperone CDC37 was transcriptionally induced by heat shock factor 1 (HSF1), which activated the CDC37 core promoter, containing an interspecies conserved heat shock element. In contrast, knockdown of CDC37 decreased EMT-coupled release of CD9-containing vesicles. Triple siRNA targeting CDC37, HSP90 alpha, and HSP90 beta was required for efficient reduction of this chaperone trio and to reduce tumorigenicity of the CRPC cells in vivo. Taken together, we define "stressome" as cellular stress-induced all secretion products, including EVs (200-500 nm), membrane-damaged vesicles and remnants, and extracellular HSP90 and GAPDH. Our data also indicated that CDC37 is crucial for the release of vesicular proteins and tumor progression in prostate cancer. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 2 2020 The Efficacy of Mirabegron for the Relief of Ureteral Stent-Related Symptoms 145 150 EN Hideo Otsuki Department of Urology, Abiko Toho Hospital Tomoya Yamasaki Department of Urology, Abiko Toho Hospital Shunsuke Hori Department of Urology, Abiko Toho Hospital Hiroshi Aoki Department of Urology, Abiko Toho Hospital Takeo Kosaka Department of Urology, Keio University Shinya Uehara Department of Urology, Kawasaki Medical School General Medical Center Kei Fujio Department of Urology, Abiko Toho Hospital Original Article 10.18926/AMO/58273 To investigate the efficacy of mirabegron for lower urinary tract symptoms in patients with an indwelling ureteral stent after ureterorenoscopic lithotripsy. This was a prospective follow-up study of 76 patients with stent-related symptoms (SRSs). Patients with upper urinary calculi who were pre-stented for > 2 weeks before lithotripsy were examined for the presence of SRSs by tests including the International Prostate Symptom Score (IPSS), OAB Symptom Score (OABSS), and urinary bother and pain measured by a Visual Analogue Scale (VAS) before lithotripsy. Mirabegron (50 mg/day) was prescribed post-lithotripsy for 2 weeks. SRSs were assessed at the time of stent removal. The IPSS scores improved significantly from 16.2 to 14.3 (p<0.001) and the IPSS-QoL scores decreased significantly from 5.0 to 4.6 (p=0.012). The OABSS scores improved significantly from 7.7 to 6.8 (p=0.006), and the urinary urgency scores (OABSS-Q3) decreased significantly from 3.24 to 2.68 (p<0.001). The number of nocturia episodes decreased significantly from 2.5 to 2.2 (p=0.045). Urinary bother and pain assessed by the VAS declined from 4.2 and 3.1 to 3.8 (p=0.15) and 2.5 (p=0.075), respectively. Mirabegron significantly improved SRSs and the number of nocturia episodes due to a ureteral stent. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 2 2020 Increased Plasma Levels of Platelet Factor 4 and β-thromboglobulin in Women with Recurrent Pregnancy Loss 115 122 EN Sayoko Kotani Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yasuhiko Kamada Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Keiko Shimizu Okayama Rosai Hospital Ai Sakamoto Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mikiya Nakatsuka Graduate School of Health Sciences, Okayama University Yuji Hiramatsu Okayama City Hospital Hisashi Masuyama Department of Obstetrics and Gynecology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/58269 Thrombosis in decidual vessels is one of the mechanisms of pregnancy loss. However, few studies have assessed the relation between platelet activation, which is known to cause of thrombosis, and recurrent pregnancy loss (RPL). We investigated platelet activation in women with RPL compared to controls by measuring plasma levels of platelet factor 4 (PF4) and β-thromboglobulin (βTG), and assessed correlations between PF4/βTG and coagulative risk factors associated with RPL. The study group included 135 women who had experienced two or more consecutive pregnancy losses. The control group included 28 age-matched healthy women who had never experienced pregnancy loss. PF4 and βTG plasma levels were significantly higher in the women with RPL than controls (PF4: 14.0 [8.0-20.0] vs. 9.0 [6.0-12.0] ng/ml, p=0.043; βTG: 42.0 [24.3-59.8] vs. 31.5 [26.6-36.4] ng/ml, p=0.002). There was a significant association between βTG and anti-phosphatidylethanolamine antibody immunoglobulin M (aPE IgM) (p=0.048). Among the women with RPL, 18 of those who were positive for PF4 (45%) and 18 of those who were positive for βTG (37%) were negative for all known coagulative risk factors associated with RPL. Measurements of PF4 and βTG may be important because they help identify women who are at risk of RPL. No potential conflict of interest relevant to this article was reported.

IOP Publishing Acta Medica Okayama 2053-1591 7 3 2020 Inhomogeneous superconductivity in thin crystals of FeSe1-xTex (x=1.0, 0.95, and 0.9) 036001 EN Ritsuko Eguchi Research Institute for Interdisciplinary Science, Okayama University Megumi Senda Graduate School of Natural Science and Technology, Okayama University Eri Uesugi Graduate School of Natural Science and Technology, Okayama University Hidenori Goto Research Institute for Interdisciplinary Science, Okayama University Akihiko Fujiwara Department of Nanotechnology for Sustainable Energy, Kwansei Gakuin University Yasuhiko Imai Japan Synchrotron Radiation Research Institute (JASRI) Shigeru Kimura Japan Synchrotron Radiation Research Institute (JASRI) Takashi Noji Department of Applied Physics, Tohoku University Yoji Koike Department of Applied Physics, Tohoku University Yoshihiro Kubozono Research Institute for Interdisciplinary Science, Okayama University We investigated the temperature dependence of resistivity in thin crystals of FeSe1-xTex (x = 1.0, 0.95, and 0.9), though bulk crystals with 1.0 x 0.9 are known to be non-superconducting. With decreasing thickness of the crystals, the resistivity of x = 0.95 and 0.9 decreases and reaches zero at a low temperature, which indicates a clear superconducting transition. The anomaly of resistivity related to the structural and magnetic transitions completely disappears in 55- to 155-nm-thick crystals of x = 0.9, resulting in metallic behavior in the normal state. Microbeam x-ray diffraction measurements were performed on bulk single crystals and thin crystals of FeSe1-xTex. A significant difference of the lattice constant, c, was observed in FeSe1-xTex, which varied with differing Te content (x), and even in crystals with the same x, which was mainly caused by inhomogeneity of the Se/Te distribution. It has been found that the characteristic temperatures causing the structural and magnetic transition (T-t), the superconducting transition (T-c), and the zero resistivity (T-c(zero)) are closely related to the value of c in thin crystals of FeSe1-xTex. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 1 2020 Metabolic Profiling of the Cerebrospinal Fluid in Pediatric Epilepsy 65 72 EN Tomoyuki Akiyama Department of Child Neurology, Okayama University Hospital Daisuke Saigusa Tohoku Medical Megabank Organization, Tohoku University Yuki Hyodo Department of Child Neurology, Okayama University Hospital Keiko Umeda Tohoku Medical Megabank Organization, Tohoku University Reina Saijo Tohoku Medical Megabank Organization, Tohoku University Seizo Koshiba Tohoku Medical Megabank Organization, Tohoku University Katsuhiro Kobayashi Department of Child Neurology, Okayama University Hospital Original Article 10.18926/AMO/57955 To characterize metabolic profiles within the central nervous system in epilepsy, we performed gas chromatography-tandem mass spectrometry (GC-MS/MS)-based metabolome analysis of the cerebrospinal fluid (CSF) in pediatric patients with and without epilepsy. The CSF samples obtained from 64 patients were analyzed by GC-MS/MS. Multivariate analyses were performed for two age groups, 0-5 years of age and 6-17 years of age, to elucidate the effects of epilepsy and antiepileptic drugs on the metabolites. In patients aged 0-5 years (22 patients with epilepsy, 13 without epilepsy), epilepsy patients had reduced 2-ketoglutaric acid and elevated pyridoxamine and tyrosine. In patients aged 6-17 years (12 with epilepsy, 17 without epilepsy), epilepsy patients had reduced 1,5-anhydroglucitol. Valproic acid was associated with elevated 2-aminobutyric acid, 2-ketoisocaproic acid, 4-hydroxyproline, acetylglycine, methionine, N-acetylserine, and serine. Reduced energy metabolism and alteration of vitamin B6 metabolism may play a role in epilepsy in young children. The roles of 1,5-anhydroglucitol in epilepsy in older children and in levetiracetam and zonisamide treatment remain to be explained. Valproic acid influenced the levels of amino acids and related metabolites involved in the metabolism of serine, methionine, and leucine. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 20 5 2019 Targeting Ovarian Cancer Cells Overexpressing CD44 with Immunoliposomes Encapsulating Glycosylated Paclitaxel 1042 EN Apriliana Cahya Khayrani Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hafizah Mahmud Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Maram H. Zahra Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Aung Ko Ko Oo Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Miharu Oze Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Juan Du Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Md Jahangir Alam Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Said M. Afify Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hagar A. Abu Quora Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Tsukasa Shigehiro Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Anna Sanchez Calle Division of Molecular and Cellular Medicine, National Cancer Center Research Institute Nobuhiro Okada Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Akimasa Seno Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Koki Fujita Ensuiko Sugar Refining Co., Ltd. Hiroki Hamada Faculty of Science, Okayama University of Science Yuhki Seno Graduate School of Pharmaceutical Science, Tokushima University Tadakatsu Mandai Faculty of Life Science, Kurashiki University of Science and the Arts Masaharu Seno Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Paclitaxel (PTX) is one of the front-line drugs approved for the treatment of ovarian cancer. However, the application of PTX is limited due to the significant hydrophobicity and poor pharmacokinetics. We previously reported target-directed liposomes carrying tumor-selective conjugated antibody and encapsulated glycosylated PTX (gPTX-L) which successfully overcome the PTX limitation. The tubulin stabilizing activity of gPTX was equivalent to that of PTX while the cytotoxic activity of gPTX was reduced. In human ovarian cancer cell lines, SK-OV-3 and OVK18, the concentration at which cell growth was inhibited by 50% (IC50) for gPTX range from 15⁻20 nM, which was sensitive enough to address gPTX-L with tumor-selective antibody coupling for ovarian cancer therapy. The cell membrane receptor CD44 is associated with cancer progression and has been recognized as a cancer stem cell marker including ovarian cancer, becoming a suitable candidate to be targeted by gPTX-L therapy. In this study, gPTX-loading liposomes conjugated with anti-CD44 antibody (gPTX-IL) were assessed for the efficacy of targeting CD44-positive ovarian cancer cells. We successfully encapsulated gPTX into liposomes with the loading efficiency (LE) more than 80% in both of gPTX-L and gPTX-IL with a diameter of approximately 100 nm with efficacy of enhanced cytotoxicity in vitro and of convenient treatment in vivo. As the result, gPTX-IL efficiently suppressed tumor growth in vivo. Therefore gPTX-IL could be a promising formulation for effective ovarian cancer therapies. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 9 7 2019 Influence of Numerical Aperture on Molten Area Formation in Fusion Micro-Welding of Glass by Picosecond Pulsed Laser 1412 EN Zhiyong Ouyang Graduate School of Natural Science and Technology, Okayama University Yasuhiro Okamoto Graduate School of Natural Science and Technology, Okayama University Yuta Ogino Graduate School of Natural Science and Technology, Okayama University Tomokazu Sakagawa Akira Okada Graduate School of Natural Science and Technology, Okayama University Focusing condition such as numerical aperture (N.A.) has a great influence on the creation of molten area and the stable welding process in fusion micro-welding of glass. In this study, a picosecond pulsed laser of 1064 nm in wavelength and 12.5 ps in pulse duration was tightly focused inside a borosilicate glass using objective lenses of numerical apertures 0.45, 0.65, and 0.85 with spherical aberration correction. Influence of numerical aperture on molten area formation was experimentally investigated through analysis of focusing situation in glass, and movement of absorption point, and then molten area characteristics were discussed. It is concluded that N.A. of 0.65 with superior focusing characteristics can form a large and continuous molten area without cracks, which enables achievement of stable joining of glass material by picosecond pulsed laser. No potential conflict of interest relevant to this article was reported.

American Chemical Society Acta Medica Okayama 00062960 58 26 2019 Photochemical Characterization of a New Heliorhodopsin from the Gram-Negative Eubacterium Bellilinea caldifistulae (BcHeR) and Comparison with Heliorhodopsin-48C12 2934 2943 EN Atsushi Shibukawa Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , Okayama University Keiichi Kojima Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , Okayama University Yu Nakajima Atmosphere and Ocean Research Institute , The University of Tokyo Yosuke Nishimura Atmosphere and Ocean Research Institute , The University of Tokyo Susumu Yoshizawa Atmosphere and Ocean Research Institute , The University of Tokyo Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , Okayama University Many microorganisms express rhodopsins, pigmented membrane proteins capable of absorbing sunlight and harnessing that energy for important biological functions such as ATP synthesis and phototaxis. Microbial rhodopsins that have been discovered to date are categorized as type-1 rhodopsins. Interestingly, researchers have very recently unveiled a new microbial rhodopsin family named the heliorhodopsins, which are phylogenetically distant from type-1 rhodopsins. Among them, only heliorhodopsin-48C12 (HeR-48C12) from a Gram-positive eubacterium has been photochemically characterized [Pushkarev, A., et al. (2018) Nature 558, 595-599]. In this study, we photochemically characterize a purple-colored heliorhodopsin from Gram-negative eubacterium Bellilinea caldifistulae (BcHeR) as a second example and identify which properties are or are not conserved between BcHeR and HeR-48C12. A series of photochemical measurements revealed several conserved properties between them, including a visible absorption spectrum with a maximum at around 550 nm, the lack of ion-transport activity, and the existence of a second-order O-like intermediate during the photocycle that may activate an unidentified biological function. In contrast, as a property that is not conserved, although HeR-48C12 shows the light adaptation state of retinal, BcHeR showed the same retinal configuration under both dark- and light-adapted conditions. These comparisons of photochemical properties between BcHeR and HeR-48C12 are an important first step toward understanding the nature and functional role of heliorhodopsins. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 5 2019 Feasibility of Laparoscopic Radical Cystectomy in Elderly Patients: A Comparative Analysis of Clinical Outcomes in a Single Institution 417 418 EN Yutaka Yanagihara Department of Urology, Ehime University Medical School Keigo Nishida Department of Urology, Ehime University Medical School Ryuta Watanabe Department of Urology, Ehime University Medical School Kanae Koyama Department of Urology, Ehime University Medical School Yuichiro Sawada Department of Urology, Ehime University Medical School Terutaka Noda Seiji Asai Department of Urology, Ehime University Medical School Tetsuya Fukumoto Department of Urology, Ehime University Medical School Noriyoshi Miura Department of Urology, Ehime University Medical School Yuki Miyauchi Department of Urology, Ehime University Medical School Tadahiko Kikugawa Department of Urology, Ehime University Medical School Takashi Saika Department of Urology, Ehime University Medical School Original Article 10.18926/AMO/57371 Laparoscopic radical cystectomy (LRC) is a standard surgical treatment for muscle-invasive bladder cancer and high-risk non-muscle-invasive bladder cancer. LRC is a less invasive modality than conventional open surgery. Therefore, even elderly patients with invasive bladder cancer may be candidates for LRC. In this study, a comparative analysis of perioperative/oncological outcomes between elderly patients and younger patients who underwent LRC was performed to assess the feasibility of LRC in elderly patients. Sixty-eight consecutive patients who underwent LRC between October 2013 and March 2018 were enrolled and stratified into those younger than 75 years (n=37) and those ≥ 75 years old (n=31). The median follow-up period was 28.2 months. The preoperative and operative parameters and complications were similar in both groups. The 2-year overall survival (OS) was 64.4% in the younger vs. 76.4% in the elderly group (p=0.053), cancer-specific survival (CSS) was 79.3% vs. 81.7% (p=0.187), and recurrence-free survival (RFS) was 58.2% vs. 75.7% (p=0.174), respectively. No significant differences were observed in OS, CSS, or RFS between the groups. No significant differences were found between the groups with respect to peri-surgical/oncological outcomes. We conclude that LRC is feasible in elderly patients. No potential conflict of interest relevant to this article was reported.

Elsevier B.V Acta Medica Okayama 24055808 19 2019 Localization of the ultraviolet-sensor Opn5m and its effect on myopia-related gene expression in the late-embryonic chick eye 100665 EN Mutsuko Kato Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Keita Sato Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Munenori Habuta Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hirofumi Fujita Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tetsuya Bando Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yuki Morizane Department of Ophthalmology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science Fumio Shiraga Department of Ophthalmology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science Satoru Miyaishi Department of Legal Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hideyo Ohuchi Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Recent studies show that exposure to ultraviolet (UV) light suppresses ocular elongation, which causes myopia development. However, the specific mechanisms of this process have not been elucidated. A UV-sensor, Opsin 5 (Opn5) mRNA was shown to be present in extraretinal tissues. To test the possibility that UV-signals mediated by Opn5 would have a direct effect on the outer connective tissues of the eye, we first examined the expression patterns of a mammalian type Opn5 (Opn5m) in the late-embryonic chicken eye. Quantitative PCR showed Opn5m mRNA expression in the cornea and sclera. The anti-Opn5m antibody stained a small subset of cells in the corneal stroma and fibrous sclera. We next assessed the effect of UV-A (375 nm) irradiation on the chicken fibroblast cell line DF-1 overexpressing chicken Opn5m. UV-A irradiation for 30 min significantly increased the expression of Early growth response 1 (Egr1), known as an immediate early responsive gene, and of Matrix metalloproteinase 2 (Mmp2) in the presence of retinal chromophore 11-cis-retinal. In contrast, expression of Transforming growth factor beta 2 and Tissue inhibitor of metalloproteinase 2 was not significantly altered. These results indicate that UV-A absorption by Opn5m can upregulate the expression levels of Egr1 and Mmp2 in non-neuronal, fibroblasts. Taken together with the presence of Opn5m in the cornea and sclera, it is suggested that UV-A signaling mediated by Opn5 in the extraretinal ocular tissues could influence directly the outer connective tissues of the chicken late-embryonic eye. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 4 2019 Evaluation of Treatment Response in Prostate Cancer and Renal Cell Carcinoma Patients Using 11C-choline PET/CT Findings 341 347 EN Kazuhiro Kitajima Department of Radiology, Division of Nuclear Medicine and PET Center, Hyogo College of Medicine Shingo Yamamoto Department of Urology, Hyogo College of Medicine Yukako Nakanishi Department of Urology, Hyogo College of Medicine Yusuke Yamada Department of Urology, Hyogo College of Medicine Takahiko Hashimoto Department of Urology, Hyogo College of Medicine Toru Suzuki Department of Urology, Hyogo College of Medicine Shuken Go Department of Urology, Hyogo College of Medicine Akihiro Kanematsu Department of Urology, Hyogo College of Medicine Michio Nojima Department of Urology, Hyogo College of Medicine Masayuki Fujiwara Department of Radiology, Hyogo College of Medicine Hayato Kaida Department of Radiology, Kindai University Faculty of Medicine Masakatsu Tsurusaki Department of Radiology, Kindai University Faculty of Medicine Tomonori Kanda Department of Radiology, Kobe University Graduate School of Medicine Yukihisa Tamaki Department of Radiation Oncology, Shimane University School of Medicine Koichiro Yamakado Department of Radiology, Hyogo College of Medicine Original Article 10.18926/AMO/56936 We investigated the effectiveness of 11C-choline-positron emission tomography/computed tomography (PET/CT) for evaluating treatment response in patients with prostate cancer or renal cell carcinoma. We performed 34 11C-choline PET/CT scans before/after a combined total of 17 courses of treatment in 6 patients with prostate cancer and 2 with renal cell carcinoma. The 17 treatments including hormonal therapy, radiotherapy, chemotherapy, radium-223, molecular target therapy, radiofrequency ablation, transcatheter arterial embolization, and cancer immunotherapy yielded 1 (5.9%) complete metabolic response (CMR), 3 (17.6%) partial metabolic responses (PMRs), 2 (11.8%) stable metabolic diseases (SMDs), and 11 (64.7%) progressive metabolic diseases (PMDs). Target lesions were observed in bone (n=14), lymph nodes (n=5), lung (n=2), prostate (n=2), and pleura (n=1), with CMR in 4, PMR in 10, SMD in 8 and PMD in 2 lesions. SUVmax values of the target lesions before and after treatment were 7.87±2.67 and 5.29±3.98, respectively, for a mean reduction of −35.4±43.6%. The response for the 8 prostate cancer-treatment courses was PMD, which correlated well with changes in serum prostatic specific antigen (PSA) (7 of 8 cases showed increased PSA). 11C-choline-PET/CT may be an effective tool for detecting viable residual tumors and evaluating treatment response in prostate cancer and renal cell carcinoma patients. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 4 2019 Construction and Characterization of a PGN_0297 Mutant of Porphyromonas gingivalis: Evidence of the Contribution of PGN_0297 to Gingipain Activity 315 323 EN Shintaro Ono Department of Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masaaki Nakayama Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Masato Tachibana Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Abu Saleh Muhammad Shahriar Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Wang Heling Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shogo Takashiba Department of Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Naoya Ohara Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/56933 The periodontal pathogen Porphyromonas gingivalis shows colonial pigmentation on blood agar and produces gingipains (Kgp, RgpA, and RgpB), cysteine proteases involved in an organism’s virulence and pigmentation. We showed previously that deletion of the PGN_0300 gene abolished the pigmentation activity and reduced the proteolytic activity of gingipains. The role of the PGN_0297 gene, which consists of an operon with the PGN_0300 gene, is unclear. Herein we examined the effect of PGN_0297 gene deletion on the pigmentation and proteolytic activities and transcriptional levels of gingipains. A PGN_0297 gene deletion mutant (ΔPGN_0297) did not exhibit the pigmentation. The proteolytic activity of the gingipains was decreased in the culture supernatant and on the cell surface of ΔPGN_0297. The mutant ΔPGN_0297 failed to attenuate Akt phosphorylation at Thr308 and Ser473, but both phosphorylations were attenuated in the wild-type and its complementation strain. The deletion of PGN_0297 gene did not substantially affect the transcriptional levels of the gingipain genes kgp, rgpA, and rgpB. Taken together, these results indicate that PGN_0297 is closely involved in the secretion and maturation of gingipains. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 4 2019 Dynamic Reorganization of Microtubule and Glioma Invasion 285 297 EN Yoshihiro Otani Department of Neurosurgery, The University of Texas Health Science Center at Houston Tomotsugu Ichikawa Department of Neurosurgery, Kagawa Prefectural Central Hospital Kazuhiko Kurozumi Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Isao Date Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Review 10.18926/AMO/56930 Gliomas are characterized as highly diffuse infiltrating tumors, and currently available treatments such as surgery, radiation and chemotherapy are unfeasible or show limited efficacy against these tumors. Recent genetic and epigenetic analyses of glioma have revealed increasing evidence of the role of driver genetic alterations in glioma development and led to the identification of prognostic factors. Despite these findings, the survival rates of glioma patients remain low, and alternative treatments and novel targets are needed. Recent studies identified neural stem cells as the possible origin of gliomas, and some evidence has revealed shared functions and mechanisms between glioma cells and neurons, also supporting their similarity. The cytoskeleton plays important roles in the migration of normal cells as well as cancer cells. Recent reports have described a role for microtubules, a component of the cytoskeleton, in glioma invasion. Notably, several factors that regulate microtubule functions, such as microtubule-associated proteins, plus-end tracking proteins, or motor proteins, are upregulated in glioma tissues compared with normal tissue, and upregulation of these factors is associated with high invasiveness of glioma cells. In this review, we describe the mechanism of microtubules in glioma invasion and discuss the possibility of microtubule-targeted therapy to inhibit glioma invasion. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 3 2019 Anti-N-Methyl-D-Aspartate Receptor Encephalitis in Psychiatry 189 195 EN Shinji Sakamoto Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroki Kawai Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yuko Okahisa Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ko Tsutsui Department of Neuropsychiatry, Akita University Graduate School of Medicine Takashi Kanbayashi Department of Neuropsychiatry, Akita University Graduate School of Medicine Keiko Tanaka Department of Animal Model Development, Brain Research Institute, Niigata University Yutaka Mizuki Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Manabu Takaki Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Norihito Yamada Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Review 10.18926/AMO/56860 Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a recently-discovered autoimmune disorder in which antibodies target NMDAR in the brain. The number of reported cases of anti-NMDAR encephalitis has increased rapidly. Anti-NMDAR encephalitis can be mistakenly diagnosed as psychiatric disorders because many patients present with prominent psychiatric symptoms and visit psychiatric institutions first. Thus, psychiatrists should cultivate a better understanding of anti-NMDAR encephalitis. In this review, we present the mechanisms, epidemiology, symptoms and clinical course, diagnostic tests, treatment and outcomes of patients with anti-NMDAR encephalitis. Furthermore, we discuss the diversity of clinical spectra of anti-NMDAR encephalitis, and demonstrate a differential diagnosis of psychiatric disease from the perspective of psychiatry. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 2 2019 Thyroid Function Decline and Diet in Female High School Long-distance Runners 127 133 EN Yukari Iwasaki Graduate School of Health Sciences, Okayama University Kimiko Miyahara Faculty of Health Care, Department of Nutrition, Kiryu University Nobuyuki Miyatake Department of Hygiene, Faculty of Medicine, Kagawa University Mikiya Nakatsuka Graduate School of Health Sciences, Okayama University Original Article 10.18926/AMO/56648 We aimed to clarify the state of thyroid function in female high school long-distance runners. We evaluated the associations between thyroid function and menstrual condition, bone mineral density (BMD), nutritious status, and body composition. The subjects’ height and weight were measured, along with fat percentage, fat mass, muscle mass, and BMD with dual-energy X-ray absorptiometry. A nutrition and dietary survey measured the subjects’ intake of energy and nutrients based on meals provided at the subjects’ dorm for 3 days in July of 2016 and 2017. Blood parameters including thyroid hormone and estradiol were measured. Most of the subjects (81.3%) were underweight (body mass index <18.5). The thyroid hormone free T3 value was decreased, but TSH was not increased and was similar to that observed in individuals with anorexia nervosa. In our subjects, thyroid hormone was associated with BMD and nutritional intake. To improve the menstruation abnormality of female athletes and to increase their bone density, the athletes’ weight should be managed by proper nutrient intake and the maintenance of their thyroid function. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 2 2019 Factors Affecting the Absorption of Midazolam to the Extracorporeal Membrane Oxygenation Circuit 101 107 EN Atsuyoshi Iida Department of Emergency and Critical Care and Disaster Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiromichi Naito Department of Emergency and Critical Care and Disaster Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takashi Yorifuji Department of Human Ecology, Okayama University Graduate School of Environmental and Life Science Yoshito Zamami Department of Clinical Pharmacy, Institute of Biomedical Sciences, Tokushima University Graduate School Akane Yamada Department of Pharmacy, Okayama University Hospital Tadashi Koga Drug Safety Research Laboratories, Shin Nippon Biomedical Laboratories, Ltd. Toru Imai Department of Pharmacy, Nihon University Itabashi Hospital Toshiaki Sendo Department of Pharmacy, Okayama University Hospital Atsunori Nakao Department of Emergency and Critical Care and Disaster Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shingo Ichiba Department of Surgical Intensive Care Medicine, Nippon Medical School Hospital Original Article 10.18926/AMO/56645 Sedatives are administered during extracorporeal membrane oxygenation (ECMO) therapy to ensure patient safety, reduce the metabolic rate and correct the oxygen supply-demand balance. However, the concentrations of sedatives can be decreased due to absorption into the circuit. This study examined factors affecting the absorption of a commonly used sedative, midazolam (MDZ). Using multiple ex vivo simulation models, three factors that may influence MDZ levels in the ECMO circuit were examined: polyvinyl chloride (PVC) tubing in the circuit, use of a membrane oxygenator in the circuit, and heparin coating of the circuit. We also assessed changes in drug concentration when MDZ was re-injected in a circuit. The MDZ level decreased to approximately 60% of the initial concentration in simulated circuits within the first 30 minutes. The strongest factor in this phenomenon was contact with the PVC tubing. Membrane oxygenator use tended to increase MDZ loss, whereas heparin circuit coating had no influence on MDZ absorption. Similar results were obtained when a second dose of MDZ was injected to the second-use circuits. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 1 2019 Efficacy of Oral Care Provided by Interprofessional Collaboration for a Patient with Esophageal Cancer Associated with Post-polio Syndrome during Neoadjuvant Chemotherapy 71 76 EN Keiko Takahashi-Arimasa Dental Hygienist Team, Division of Medical Technology, Okayama University Hospital Reiko Kohno-Yamanaka Division of Hospital Dentistry, Central Clinical Department, Okayama University Hospital Yoshihiko Soga Division of Hospital Dentistry, Central Clinical Department, Okayama University Hospital Rumi Miura Dental Hygienist Team, Division of Medical Technology, Okayama University Hospital Manabu Morita Department of Preventive Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Case Report 10.18926/AMO/56461 Preoperative oral care is helpful to prevent postoperative complications in patients who are undergoing esophagectomy. Here, we report the case of an 81-year-old Japanese man with an upper limb disability caused by post-polio syndrome who was receiving neoadjuvant chemotherapy for esophageal cancer. He had poor oral health status and developed oral complications as a side effect of chemotherapy. He could not brush his teeth by himself. However, infection control by oral care provided by an interprofessional collaboration successfully improved his oral hygiene, and his follow-up involved no severe complications. Interprofessional collaboration is useful especially for patients with upper limb disability. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 1 2019 Reduction of Postoperative Pain by Addition of Intravenous Acetaminophen after Total Hip Arthroplasty: A Retrospective Cohort Study 7 14 EN Norio Fukumori Community Medical Support Institute, Faculty of Medicine, Saga University Motoki Sonohata Department of Orthopaedic Surgery, Faculty of Medicine, Saga University Masaru Kitajima Department of Orthopaedic Surgery, Faculty of Medicine, Saga University Shunsuke Kawano Department of Orthopaedic Surgery, Faculty of Medicine, Saga University Tsuyoshi Kurata Community Medical Support Institute, Faculty of Medicine, Saga University Yuta Sakanishi Community Medical Support Institute, Faculty of Medicine, Saga University Takashi Sugioka Community Medical Support Institute, Faculty of Medicine, Saga University Masaaki Mawatari Department of Orthopaedic Surgery, Faculty of Medicine, Saga University Original Article 10.18926/AMO/56453 We evaluated the analgesic effects of multimodal pain control in which intravenous acetaminophen (IV APAP) was added to the standard protocol for Japanese patients who had undergone a total hip arthroplasty (THA). We performed a retrospective cohort study of 180 patients aged 66.4±10.5 years (30% male) who had undergone a THA (Oct. 2014 to Feb. 2015) at our hospital. The control patients were administered the standard analgesic protocol: flurbiprofen axetil as a continuous intravenous infusion and oral celecoxib (NAPAP; n=109). The patients in the new analgesic protocol group received IV APAP in addition to the standard analgesic protocol (APAP; n=71). The primary outcome was the maximum value of postoperative pain the patients reported on a numerical rating scale (NRS) during the first 24 h post-surgery. A univariate analysis and multivariate analyses adjusted for age, sex, the stage of hip osteoarthritis, preoperative pain, and surgical time showed that the maximum postoperative pain NRS scores during the first 24 h after surgery was significantly lower when the APAP protocol was used. The addition of IV APAP to the current standard multimodal analgesia protocol for Japanese patients who have undergone a THA may decrease the patients’ postoperative pain. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 73 1 2019 The Role of Kallikrein-Related Peptidases in Atopic Dermatitis 1 6 EN Shin Morizane Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Review 10.18926/AMO/56452 Excessive protease activity is a characteristic abnormality that affects the epidermal barrier in patients with atopic dermatitis (AD). Kallikrein-related peptidases (KLKs) are excessively expressed in AD lesions, and it is suggested that the abnormal action of KLKs is involved in the skin barrier dysfunction in AD. In other words, overexpressed KLKs disrupt the normal barrier function, and due to that breakdown, external substances that can become antigens of AD easily invade the epidermis, resulting in dermatitis, coupled with the induction of Th2 cytokines. Further investigations are required to elucidate the role of KLKs in AD; this knowledge could contribute to the design of new therapeutic and prophylactic drugs for AD. No potential conflict of interest relevant to this article was reported.

Biophysical Society Acta Medica Okayama 00063495 115 1 2018 Retinal Configuration of ppR Intermediates Revealed by Photoirradiation Solid-State NMR and DFT 72 83 EN Yoshiteru Makino Graduate School of Engineering, Yokohama National University Izuru Kawamura Graduate School of Engineering, Yokohama National University Takashi Okitsu Laboratory of Organic Chemistry for Life Science, Kobe Pharmaceutical University Akimori Wada Laboratory of Organic Chemistry for Life Science, Kobe Pharmaceutical University Naoki Kamo Faculty of Advanced Life Science, Hokkaido University Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Kazuyoshi Ueda Graduate School of Engineering, Yokohama National University Akira Naito Graduate School of Engineering, Yokohama National University Pharanois phoborhodopsin (ppR) from Natronomonas pharaonis is a transmembrane photoreceptor protein involved in negative phototaxis. Structural changes in ppR triggered by photoisomerization of the retinal chromophore are transmitted to its cognate transducer protein (pHtrII) through a cyclic photoreaction pathway involving several photointermediates. This pathway is called the photocycle. It is important to understand the detailed configurational changes of retinal during the photocycle. We previously observed one of the photointermediates (M-intermediates) by in situ photoirradiation solid-state NMR experiments. In this study, we further observed the 13C cross-polarization magic-angle-spinning NMR signals of late photointermediates such as O- and N'-intermediates by illumination with green light (520 nm). Under blue-light (365 nm) irradiation of the M-intermediates, 13C cross-polarization magic-angle-spinning NMR signals of 14- and 20-13C-labeled retinal in the O-intermediate appeared at 115.4 and 16.4 ppm and were assigned to the 13-trans, 15-syn configuration. The signals caused by the N'-intermediate appeared at 115.4 and 23.9 ppm and were assigned to the 13-cis configuration, and they were in an equilibrium state with the O-intermediate during thermal decay of the M-intermediates at -60°C. Thus, photoirradiation NMR studies revealed the photoreaction pathways from the M- to O-intermediates and the equilibrium state between the N'- and O-intermediate. Further, we evaluated the detailed retinal configurations in the O- and N'-intermediates by performing a density functional theory chemical shift calculation. The results showed that the N'-intermediate has a 63° twisted retinal state due to the 13-cis configuration. The retinal configurations of the O- and N'-intermediates were determined to be 13-trans, 15-syn, and 13-cis, respectively, based on the chemical shift values of [20-13C] and [14-13C] retinal obtained by photoirradiation solid-state NMR and density functional theory calculation. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2041-1723 7 2016 Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs 12501 EN Takashi Ohtsuki Department of Biomedical Engineering, Okayama University Shigeto Kanzaki Department of Biomedical Engineering, Okayama University Sae Nishimura Department of Biomedical Engineering, Okayama University Yoshio Kunihiro Department of Biomedical Engineering, Okayama University Masahiko Sisido Department of Biomedical Engineering, Okayama University Kazunori Watanabe Department of Biomedical Engineering, Okayama University The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4 h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at ∼405 nm at 125 mW cm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19 s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells. No potential conflict of interest relevant to this article was reported.

American Chemical Society Acta Medica Okayama 00027863 139 12 2017 Demonstration of a Light-Driven SO42- Transporter and Its Spectroscopic Characteristics. 4376 4389 EN Akiko Niho Faculty of Pharmaceutical Sciences, Okayama University Susumu Yoshizawa Atmosphere and Ocean Research Institute, The University of Tokyo Takashi Tsukamoto Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Marie Kurihara Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Shinya Tahara Molecular Spectroscopy Laboratory, RIKEN Yu Nakajima Atmosphere and Ocean Research Institute, The University of Tokyo Misao Mizuno Department of Chemistry, Graduate School of Science, Osaka University Hikaru Kuramochi Ultrafast Spectroscopy Research Team, RIKEN Center for Advanced Photonics Tahei Tahara Ultrafast Spectroscopy Research Team, RIKEN Center for Advanced Photonics Yasuhisa Mizutani Department of Chemistry, Graduate School of Science, Osaka University Yuki Sudo Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University In organisms, ion transporters play essential roles in the generation and dissipation of ion gradients across cell membranes. Microbial rhodopsins selectively transport cognate ions using solar energy, in which the substrate ions identified to date have been confined to monovalent ions such as H+, Na+, and Cl-. Here we report a novel rhodopsin from the cyanobacterium Synechocystis sp. PCC 7509, which inwardly transports a polyatomic divalent sulfate ion, SO42-, with changes of its spectroscopic properties in both unphotolyzed and photolyzed states. Upon illumination, cells expressing the novel rhodopsin, named Synechocystis halorhodopsin (SyHR), showed alkalization of the medium only in the presence of Cl- or SO42-. That alkalization signal was enhanced by addition of a protonophore, indicating an inward transport of Cl- and SO42- with a subsequent secondary inward H+ movement across the membrane. The anion binding to SyHR was suggested by absorption spectral shifts from 542 to 536 nm for Cl- and from 542 to 556 nm for SO42-, and the affinities of Cl- and SO42- were estimated as 0.112 and 5.81 mM, respectively. We then performed time-resolved spectroscopic measurements ranging from femtosecond to millisecond time domains to elucidate the structure and structural changes of SyHR during the photoreaction. Based on the results, we propose a photocycle model for SyHR in the absence or presence of substrate ions with the timing of their uptake and release. Thus, we demonstrate SyHR as the first light-driven polyatomic divalent anion (SO42-) transporter and report its spectroscopic characteristics. No potential conflict of interest relevant to this article was reported.

Elsevier B.V. Acta Medica Okayama 1742-7061 57 2017 Biocompatible nanostructured solid adhesives for biological soft tissues 404 413 EN Masahiro Okada Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Akira Nakai Department of Biomaterials, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Satoshi Hara Emilio Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Tetsushi Taguchi Polymeric Biomaterials Group, RCFM, National Institute for Materials Science Takayoshi Nakano Division of Materials and Manufacturing Science, Graduate School of Engineering, Osaka University Takuya Matsumoto Department of Biomaterials, Okayama University Over the past few years, the development of novel adhesives for biological soft tissue adhesion has gained significant interest. Such adhesives should be non-toxic and biocompatible. In this study, we synthesized a novel solid adhesive using nanostructured hydroxyapatite (HAp) and evaluated its physical adhesion properties through in vitro testing with synthetic hydrogels and mouse soft tissues. The results revealed that HAp-nanoparticle dispersions and HAp-nanoparticle-assembled nanoporous plates showed efficient adhesion to hydrogels. Interestingly, the HAp plates showed different adhesive properties depending upon the shape of their nanoparticles. The HAp plate made up of 17 nm-sized nanoparticles showed an adhesive strength 2.2 times higher than that of the conventional fibrin glue for mouse skin tissues. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2041-1723 7 2016 Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs 12501 12501 EN Takashi Ohtsuki Department of Biomedical Engineering, Okayama University Shigeto Kanzaki Department of Biomedical Engineering, Okayama University Sae Nishimura Department of Biomedical Engineering, Okayama University Yoshio Kunihiro Department of Biomedical Engineering, Okayama University Masahiko Sisido Department of Biomedical Engineering, Okayama University Kazunori Watanabe Department of Biomedical Engineering, Okayama University The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4 h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at ∼405 nm at 125 mW cm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19 s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 128 1 2016 薬物相互作用 (35―オピオイド鎮痛薬の薬物相互作用） 53 59 EN Hikaru Sada Makoto Kajizono Yoshihisa Kitamura Toshiaki Sendo No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 128 1 2016 原発性十二指腸粘液癌の一例― 本邦報告16例の検討― 21 25 EN Ikumi Hamano Yusuke Matsumoto Yoshikatsu Endo Naoki Watanabe Kyouhei Kai Shizou Sato Yoji Wani 　Primary mucinous adenocarcinoma of the duodenum is rare. Here we report a case we recently encountered, and we review 16 cases reported in Japan. An 82-year-old Japanese woman was admitted to our hospital complaining of abdominal pain and heartburn. An endoscopic examination revealed a Type 2 tumor in the descending limb of the duodenum, and endoscopically obtained specimens revealed a poorly differentiated adenocarcinoma. We performed a curative pancreatoduodenectomy with lymph node resection, and the surgical specimen revealed that the duodenum was the primary site of the mucinous adenocarcinoma. The patient is currently alive ＞ 1 year after the operation without any evidence of recurrence. Of the 16 patients reviewed, all patients had advanced tumors those depth were T3-T4. 9 patients had lymph node metastasis and 4 patients had peritoneal dissemination at the time of surgery. Since mucinous adenocarcinoma of the duodenum is often progressive cancer at a diagnosis, which is tend to have a worse prognosis than other histological types. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2015 Modification of the cell adhesion and hydrophilic characteristics of poly (ether-ether-ketone) by 172-nm Xenon excimer radiation EN Yukimasa Okada No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0029-0203 95 10 1991 網膜内境界膜の超微細構造 第2報 ポリエチレンイミンによる網膜内境界膜･陰性荷電部位の観察 951 958 EN 陽イオントレーサーのPEIによる浸漬法を用いて,家兎の網膜内境界膜の陰性荷電部位(anionic sites)の透過型電子顕微鏡で観察した.1)この方法は手技が簡単で,網膜内境界膜のanionic sitesを正確にかつ短時間に観る上で有効である.2)網膜後極部(視放部,髄翼部)ではPEI粒子が網膜内境界膜の透明層中にほぼ均等に付着していた.3) PEI粒子の直径は平均20 nmで,40〜50 nmの間隔で並んでおり,透明層の細線維の間隔と一致した.4)網膜周辺部では透明層中のPEI粒子の分布は疎で,不均一に付着していた.5)眼球の全部位において,網膜内境界膜の緻密層にはPEI粒子の付着が観られなかった.6)残存硝子体線維にはPEI粒子が付着していた.以上網膜内境界膜のanionic sitesは荷電関門(charge barrier)として網膜硝子体間の物質移動の関門の役割を果たす No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0029-0203 93 4 1989 網膜内境界膜の超微細構造 第1報 臨界点乾燥表面レプリカ法による網膜内境界膜の観察 429 438 EN コラゲナーゼ処理と臨界点乾燥表面レプリカ法を用いてウサギの網膜内境界膜の超微細構造を透過型電子顕微鏡で観察した.1)この方法は手技が比較的簡便で,再現性が高く,広範囲,かつ鳥瞰図的に網膜内境界膜を観察できる点で優れている.2)網膜内境界膜は基底膜の特徴である網目構造をとり,篩が三次元的に重なり合って広がっていた.3)孔の直径は10〜25 nmに集中し,視放部(直径の平均値は13.43 nm),髄翼部(13.59 nm),網膜周辺部(13.40 nm)で,統計学的な部位による有意差は認められなかった.4)網膜周辺部では硝子体線維が密に残り,網膜内境界膜の透見できる部分が非常に少なく,篩の分布が疎でその形も他と多少異なっていた.これらの網膜内境界膜の篩はsize barrierとして網膜硝子体間の物質移動の関門の役割を果すと考えられる.篩の分布が疎で硝子体線維が密な網膜周辺部より網膜後極部,特に視放部で物質移動がより多く行われていると推察した No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 126 1 2014 PET/CT SUVmaxは子宮体癌の予後不良因子になりうる 11 15 EN Keiichiro Nakamura Ikuo Joja Chikako Fukushima Tomoko Haruma Chiaki Hayashi Tomoyuki Kusumoto Noriko Seki Atsushi Hongo Yuji Hiramatsu No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 125 3 2013 Th２サイトカインはアトピー性皮膚炎患者における カリクレイン７の発現と機能を増強する 217 220 EN Shin Morizane Kenshi Yamasaki Ai Kajita Kazuko Ikeda Maosheng Zhan Yumi Aoyama Richard L Gallo Keiji Iwatsuki No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 125 3 2013 NASHに対する水素分子の有用性 201 204 EN Daisuke Kawai Akinobu Takaki Kazuhide Yamamoto No potential conflict of interest relevant to this article was reported.

Elsevier Ltd Acta Medica Okayama 1063-4584 21 1 2013 Histone deacetylase inhibitors suppress mechanical stress-induced expression of RUNX-2 and ADAMTS-5 through the inhibition of the MAPK signaling pathway in cultured human chondrocytes 165 174 EN T. Saito K. Nishida T. Furumatsu A. Yoshida M. Ozawa T. Ozaki Objective: To investigate the inhibitory effects and the regulatory mechanisms of histone deacetylase (HDAC) inhibitors on mechanical stress-induced gene expression of runt-related transcription factor (RUNX)-2 and a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS)-5 in human chondrocytes. Methods: Human chondrocytes were seeded in stretch chambers at a concentration of 5 x 10(4) cells/chamber. Cells were pre-incubated with or without HDAC inhibitors (MS-275 or trichostatin A; TSA) for 12 h, followed by uniaxial cyclic tensile strain (CTS) (0.5 Hz, 10% elongation), which was applied for 30 min using the ST-140-10 system (STREX, Osaka, Japan). Total RNA was extracted and the expression of RUNX-2, ADAMTS-5, matrix metalloproteinase (MMP)-3, and MMP-13 at the mRNA and protein levels were examined by real-time polymerase chain reaction (PCR) and immunocytochemistry, respectively. The activation of diverse mitogen-activated protein kinase (MAPK) pathways with or without HDAC inhibitors during CTS was examined by western blotting. Results: HDAC inhibitors (TSA: 10 nM, MS-275: 100 nM) suppressed CTS-induced expression of RUNX-2, ADAMTS-5, and MMP-3 at both the mRNA and protein levels within 1 h. CTS-induced activation of p38 MAPK (p38), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (INK) MAPKs was downregulated by both HDAC inhibitors. Conclusion: The CTS-induced expression of RUNX-2 and ADAMTS-5 was suppressed by HDAC inhibitors via the inhibition of the MAPK pathway activation in human chondrocytes. The results of the current study suggested a novel therapeutic role for HDAC inhibitors against degenerative joint disease such as osteoarthritis. No potential conflict of interest relevant to this article was reported.

WILEY-VCH Acta Medica Okayama 01730835 34 16 2013 Discrimination of glycoproteins via two-color laser-induced fluorescence detection coupled with postcolumn derivatization in capillary electrophoresis 2316 2322 EN Ayumi Tabara Takashi Kaneta Here, we report a novel method consisting of capillary electrophoretic separation followed by two-color LIF detection with postcolumn derivatization. The method can be used to discriminate glycoproteins in a protein mixture containing both glycosylated and unglycosylated proteins. The detector permitted simultaneous measurements of two electropherograms obtained by 450 nm (diode laser) and 532 nm (Nd:YAG laser) lasers excited native proteins following postcolumn derivatization with naphthalene-2,3-dicarboxaldehyde and concanavalin A (Con A) labeled with tetramethylrhodamine (rhodamine-labeled Con A), respectively. So, a protein can be assigned as glycosylated if it shows a peak at the same migration time in both electropherograms. According to the proposed principle, in a single run we discriminated a glycosylated protein (thyroglobulin) from an unglycosylated protein (albumin) in the presence of rhodamine-labeled Con A. Because the methodology permits the simultaneous detection of native proteins and their complexes with a fluorescently labeled probe, it should have broad applicability to binding assays. No potential conflict of interest relevant to this article was reported.

Elsevier Ireland Ltd. Acta Medica Okayama 0169-5002 76 1 2012 Strong anti-tumor effect of NVP-AUY922, a novel Hsp90 inhibitor, on non-small cell lung cancer 26 31 EN Tsuyoshi Ueno Kazunori Tsukuda Shinichi Toyooka Midori Ando Munenori Takaoka Junichi Soh Hiroaki Asano Yuho Maki Takayuki Muraoka Norimitsu Tanaka Kazuhiko Shien Masashi Furukawa Tomoki Yamatsuji Katsuyuki Kiura Yoshio Naomoto Shinichiro Miyoshi The anti-tumor activity of a newly developed Hsp90 inhibitor, NVP-AUY922 (AUY922), against non-small cell lung cancer (NSCLC) was examined. Twenty-one NSCLC cell lines were used, the somatic alterations of which were characterized. Cell proliferation was analyzed using a modified MTS assay. Expression of the client proteins was assessed using Western blotting. The cell cycle was analyzed using flow cytometry. The IC50 value of AUY922 for the NSCLC cell lines ranged from 5.2 to 860 nM (median, 20.4 nM). Based on previous data, cells with an IC50 of less than 50 nM were classified as sensitive cells and 19 of the 21 NSCLC cell lines were judged to be sensitive. The IC50 of five malignant pleural mesothelioma (MPM) cell lines revealed that the MPM cells had a significantly higher IC50 value (median, 89.2 nM; range, 22.2-24, 100 nM) than the NSCLC cells (p = 0.015). There was significant depletion of both the total and phosphorylated client proteins - EGFR, MET, HERZ and ART - at low drug concentrations (50-100 nM) in drug-sensitive cell lines. Cell-cycle analysis was performed for two sensitive cell lines, H1975 and H838. Following AUY922 treatment, an increase in the sub-G(0)-G(1) cell population, as well as appearance of cleaved PARP expression, indicated the induction of apoptosis. In conclusion, AUY922 was effective against most NSCLC cell lines, independent of the type of known molecular alteration, and appears to be a promising new drug for the treatment of NSCLC. (C) 2011 Elsevier Ireland Ltd. All rights reserved. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 124 3 2012 糖尿病における移植療法の現状 249 252 EN Hirofumi Noguchi Toshiyoshi Fujiwara No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 124 3 2012 合成レチノイドAm80はTh1とTh17を抑制することにより慢性移植片対宿主病を改善する 197 201 EN Hisakazu Nishimori Yoshinobu Maeda Mitsune Tanimoto No potential conflict of interest relevant to this article was reported.

Hindawi Publishing Corporation Acta Medica Okayama 2012 2012 Theoretical Study on the Detection of Tilted Lipid Bilayers Using Surface Plasmon Resonance Techniques EN Koyo Watanabe Koji Matsuura Effective refractive indices detected using surface plasmon resonance techniques are calculated as a function of the tilt angle of lipid bilayers in a multilayered model. The changes in the effective refractive indices are derived from the shift of an excitation angle of surface plasmon. To obtain effective refractive index plots, we determined refractive index changes in the lipid bilayers with 3 and 5 nm thicknesses as a function of tilt angle and obtained a relationship between the effective refractive index and tilt angle. We also showed that the effective refractive index depended on the lipid bilayers thickness and anisotropic permittivities, which vary with interchain distance. No potential conflict of interest relevant to this article was reported.

American Institute of Physics. Acta Medica Okayama 0021-8979 109 4 2011 Spectroscopic evidence of the formation of (V,Ti)O₂ solid solution in VO₂ thinner films grown on TiO₂(001) substrates 043702-1 043702-6 EN Y. Muraoka K. Saeki R. Eguchi T. Wakita M. Hirai T. Yokoya S. Shin We have prepared VO2 thin films epitaxially grown on TiO2(001) substrates with thickness systematically varied from 2.5 to 13 nm using a pulsed laser deposition method, and studied the transport property and electronic states of the films by means of resistivity and in situ synchrotron photoemission spectroscopy (SRPES). In resistivity measurements, the 13-nm-thick film exhibits a metal-insulator transition at around 290 K on cooling with change of three orders of magnitudes in resistivity. As the film thickness decreases, the metal-insulator transition broadens and the transition temperature increases. Below 4 nm, the films do not show the transition and become insulators. In situ SRPES measurements of near the Fermi level valence band find that the electronic state of the 2.5-nm-thick film is different than that of the temperature-induced insulator phase of VO2 itself although these two states are insulating. Ti 2p core-level photoemission measurements reveal that Ti ions exist near the interface between the films and TiO2 substrates, with a chemical state similar to that in (V,Ti)O-2 solid solution. These results indicate that insulating (V,Ti)O-2 solid solution is formed in the thinner films. We propose a simple growth model of a VO2 thin film on a TiO2(001) substrate. Near the interface, insulating (V,Ti) O-2 solid solution is formed due to the diffusion of Ti ions from the TiO2 substrate into the VO2 film. The concentration of Ti in (V,Ti) O-2 is relatively high near the interface and decreases toward the surface of the film. Beyond a certain film thickness (about 7 nm in the case of the present 13-nm-thick film), the VO2 thin film without any Ti ions starts to grow. Our work suggests that developing a technique for preparing the sharp interface between the VO2 thin films and TiO2 substrates is a key issue to study the physical property of an ultrathin film of "pure" VO2, especially to examine the presence of the novel electronic state called a semi-Dirac point phase predicted by calculations. No potential conflict of interest relevant to this article was reported.

Elsevier Science BV. Acta Medica Okayama 0926-3373 102 3-4 2011 Improvement in sulfur desorption of NOX storage and reduction catalysts using a Ba–Ti composite oxide 620 626 EN Toshiyuki Tanaka Ichirou Tajima Yuichi Kato Yasushi Nishihara Hirofumi Shinjoh A Ba-Ti composite oxide was formed on a NOx storage and reduction catalyst via impregnation of a Ba-Ti precursor solution composed of H(2)O(2) added to a complex prepared using the citric acid method. The structure of the Ba-Ti composite in solution was analyzed by chemical composition analysis and FT-Raman and UV-vis spectroscopy. MM2 calculations were performed to propose its chemical structure. Both Ba and Ti together were found to form a composite molecule in the solution. Furthermore, TEM-EDX and XRD analyses of the Ba-Ti composite oxide on the catalyst prepared by impregnation with the Ba-Ti composite aqueous solution revealed that Ba and Ti in the catalyst were highly dispersed at the nm scale. The formation of the Ba-Ti composite oxide on the NSR catalyst enhanced sulfur desorption efficiency and led to high-performance NO(X) conversion as a NO(X) storage and reduction activity catalyst after desulfation treatment. It was assumed that the existence of nano-scaled Ba compounds combined with Ti was efficient for the inhibition of the sintering of barium sulfate and its facile decomposition. It was found that dispersion of Ba compounds for NO(X) storage materials using a Ba-Ti complex solution is an efficient way to improve the durability of NSR catalysts. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 123 3 2011 薬物相互作用（22―オピオイドの薬物相互作用） 239 241 EN Rieko Kawashima Hisashi Matsunaga Toshiaki Sendo No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0021-9673 1218 25 2011 Rapid determination of multidrug resistance-associated protein in cancer cells by capillary electrophoresis immunoassay 3923 3927 EN Julius Mbuna Takashi Kaneta Totaro Imasaka The adenosine triphosphate (ATP) binding-cassette (ABC) transporters are a superfamily of cellular proteins that have been partly implicated as a cause of multidrug resistance (MDR) in cancer cells. The ABC superfamily consists of P-glycoprotein, multidrug resistance-associated proteins (MRP) and breast cancer-related proteins, of which MRP is of particular interest because of its ability to efflux a broader range of substrates. Since MRP1 is the most prominent member of the MRP family, a simple technique is needed for its quantification. We developed a simple, fast (total analysis time of 3 h) capillary electrophoresis immunoassay (CEIA) for the quantification of MRP1 in cancer cells. MRP1 antibody was labeled with fluorescein isothiocyanate. The labeled antibody was incubated with the cell lysate for a fixed interval (1 h), after which the cell lysate mixture was directly injected into the capillary to separate the complex of MRP1 and its antibody from free antibody. The noncompetitive CEIA method had a limit of detection of 0.2 nM and a good linear range (1.7-14.9 x 10(4) cells), and was fairly reproducible (RSD < 10%). The results showed that two cell lines. A549 and RDES, expressed MRP1 in the absence of doxorubicin (DOX), with A549 registering a higher expression. Compared to DOX-free cancer cells, there was an acceleration of MRP1 expression during the 12 h-exposure to DOX, after which the level of expression remained nearly constant as the intracellular accumulation of DOX decreased. The results obtained in this work indicate that the developed CEIA method is useful for relative quantification of MRPs in cancer cells. No potential conflict of interest relevant to this article was reported.

Pergamon-Elsevier Science Ltd. Acta Medica Okayama 0008-6223 49 10 2011 Superhydrophilicity on nano-rough carbon surfaces achieved by hyperthermal oxygen-atom beam exposure 3388 3391 EN Hiroshi Kinoshita Takashi Nakayama Naohiro Matsumoto Nobuo Ohmae In order to investigate a method to increase hydrophilicity on nano-rough carbon surfaces, a nano-rough surface of C(60) film and an atomically flat surface of highly oriented pyrolytic graphite (HOPG) were oxidized by hyperthermal oxygen-atom beam exposure and the hydrophilicities of the surfaces were investigated. Superhydrophilicity were achieved on these exposed carbon surfaces, which had low O/C ratio of approximately 28% and surface roughness (Ra) of approximately 3 nm. The direct oxidations on sp(2) bonded carbon atoms (basal plane) of these two carbon materials by the exposure of hyperthermal O-atom beam would contribute the superhydrophilicity. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 64 5 2010 Glycocalyx Degradation in Retinal and Choroidal Capillary Endothelium in Rats with Diabetes and Hypertension 277 283 EN Fumiaki Kumase Yuki Morizane Satoshi Mohri Ippei Takasu Aiji Ohtsuka Hiroshi Ohtsuki Original Article 10.18926/AMO/40502 Endothelial glycocalyx (GCX) has been reported as a protective factor for vascular endothelial cells (VEC) in diabetes and hypertension. However, the involvement of GCX impairment in ocular vasculopathy remains unclear. We evaluated the changes in the GCX thicknesses of the retinal and choroidal capillaries in rats with diabetes and hypertension by cationic colloidal iron staining using a transmission electron microscope. In the control group, the mean (standard error of the mean) thicknesses of retinal and choroidal GCX were 60.2 (1.5) nm and 84.3 (3.1) nm, respectively. The diabetic rats showed a significant decrease of GCX thickness in the retina, but not in the choroid, compared to controls (28.3 (0.3) nm, p＜0.01 and 77.8 (1.4) nm, respectively). In the hypertensive rats, both retinal and choroidal GCX were significantly decreased compared to the control values (10.9 (0.4) nm and 13.2 (1.0) nm, respectively, both p＜0.01). Moreover, we could visualize the adhesion of leukocytes and platelets on the luminal surface of VEC, at the site where the GCX was markedly degraded. These findings suggest that the GCX prevents adhesion of leukocytes and platelets to the VEC surface, and this impairment may lead to ocular vasculopathy in diabetes and hypertension. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 73 1-2 2003 Alternative methods to evaluate the protective ability of sunscreen against photo-genotoxicity 59 66 EN Megumi Toyoshima Kanako Hosoda Miho Hanamura Keinosuke Okamoto Hiroshi Kobayashi Tomoe Negishi <p>Numerous epidemiological investigations show that sunlight is carcinogenic to humans and that the use of sunscreen may be effective in decreasing the risk of skin cancer. The biological activity of a sunscreen is evaluated by its ability to protect human skin from erythema as represented by a Sun Protection Factor (SPF). We propose that the sunscreen's protective effect against sunlight-induced genotoxicity, including mutation, should also be taken into account. In this study we examined the protective ability of sunscreens against natural sunlight and UV-induced genotoxicity in Drosophila somatic cells. We prepared three kinds of sunscreen samples, each with an SPF value of 20, 40 or 60 and compared their protective activities with commercial sunscreens. When a sunscreen of SPF 20, 40 or 60 was pasted on the plastic cover of a petri dish in which Drosophila larvae were exposed to the sun or UV lamps, genotoxicity decreased as the SPF of the sunscreen increased, relative to levels of genotoxicity observed in samples without sunscreen. However, the protective abilities of sunscreens were unexpectedly not so different from each other. To reveal the relationship between the protective activity of sunscreen and the wavelength of light with which larvae were irradiated through the sunscreen, we measured the transmittance of light through the petri dish cover on which the sunscreen was pasted. Effective protection was demonstrated by removing components of light whose wavelengths were below 315 nm. We suggest, that the measurement of anti-genotoxic activity and the determination of the wavelengths of light transmitted through the sunscreen should be an alternative method for evaluating the effectiveness of a sunscreen.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 152 10 2005 Effect of annealing and hydrogen on properties of electrodeposited platinum electrode and lead-zirconate-titanate films for ferroelectric random access memory applications 688 691 EN Kazuo Kondo Keiji Watanabe Yuji Abe Junko Haji Masaru Shimizu <p>The selection of capacitor electrode materials for the nonvolatile ferroelectric random access memory is one of the most important issues because capacitor electrical characteristics are strongly influenced by the electrode materials. The lower Pt electrode was electrodeposited on the Ti seed/Pt seed layer. Two different thicknesses of Ti seed layer (5 and 15 nm) were adopted, and lead-zirconate-titanate (PZT) was deposited on the electrodeposited Pt. The Pt crystal orientation with a 5 nm Ti seed layer is much better than that with a 15 nm Ti seed layer, and the deposited PZT shows much better crystal orientation. Due to better crystal orientation of the PZT layer in the case of a 5 nm Ti seed layer, a Pt/PZT/Pt capacitor well-saturated D-V hysteresis loop was obtained whereas little current was observed in the large electric field. With the 15 nm Ti seed layer, numerous several-mu m-sized voids formed on the lower Pt electrode surface. With the 5 nm Ti seed layer, fewer voids formed on the lower Pt electrode surface. Glow discharge spectrometry measurement with a 15 nm Ti seed layer shows much higher H intensity than that with a 5 nm Ti seed layer, and the H peak coincides with the Ti peak. The H existing in the Ti seed layer must have transmitted into the PZT layer and deteriorated the PZT crystal orientation. (c) 2005 The Electrochemical Society. </p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 34 10 2005 Polar and hydrogen-bonding effects of alcohols on the emission spectrum of styrene-triethylamine system 1109 1119 EN Shunzo Yamamoto <p>The emission spectra of styrene (ST)-triethylamine (TEA) systems were measured under steady-state illumination conditions in some THF-protic solvent mixtures. The fluorescence spectrum of the ST-TEA system in THF consists of two bands (band A at 304 nm (fluorescence of ST) and band B at 460 nm (emission from an exciplex)). The intensity of band A increased and that of band B decreased with increasing amounts of protic solvents in THF-protic solvent mixtures. The increase in the intensity of band A was explained by the decrease in the concentration of free amine owing to the hydrogen-bonding interaction (or protonation) between TEA and protic solvents. The decrease in the intensity of band B was considered to be caused by the decrease in the concentration of free amine on the addition of protic solvents and the enhanced conversion of the exciplex to an ion pair with increasing solvent polarity. The polar effect was expressed as a function of the relative permittivity of the solution.</p> No potential conflict of interest relevant to this article was reported.

Pergamon-Elsevier Science Ltd. Acta Medica Okayama 0008-6223 47 8 2009 The use of graphite oxide to produce mesoporous carbon supporting Pt, Ru, or Pd nanoparticles 2120 2124 EN Kazuma Gotoh Koji Kawabata Eiji Fujii Kunimitsu Morishige Taro Kinumoto Yuki Miyazaki Hiroyuki Ishida Mesoporous carbon having platinum, ruthenium or palladium nanoparticles on exfoliated graphene sheets were produced from graphite oxide (GO) and metal complexes. The Pt included carbon was made by heating of the intercalation compound including tetraammineplatinum (II) chloride monohydrate. Samples having Ru or Pd are producible by heating in nitrogen gas atmosphere using hexaammineruthenium (III) chloride or tetraamminepalladium (II) chloride monohydrate instead of Pt complex. The particle sizes of platinum, ruthenium, and palladium were, respectively, 1–3, 1–2, and 3–7 nm. The platinum- or palladium-containing sample showed catalytic activity for oxygen reduction. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 134 11 1987 Photoluminescence spectra and vibrational structures of the srs:ce³⁺ and srse:ce³⁺ phosphors 2932 2934 EN N Yamashita Y Michitsuji S Asano <p>The details of photoluminescence and excitation spectra of powder phosphors were obtained at various temperatures between 6 and 300 K. At 300 K, two emission bands originating from the ²T_2g(5d)→²F_7/2,²F_5/2(4f) transitions are observed at 480 and 535 nm for SrS:Ce³⁺ and at 470 and 527 nm for SrSe:Ce³⁺, respectively. In the excitation spectra, bands corresponding to the ²F_5/2(4f)→²2T_2g(5d) transition are observed at 433 (SrS:Ce³⁺) and 430 nm (SrSe:Ce³⁺), and plateaus due to the fundamental absorption of the host crystals in higher energy region. The vibrational structures on the emission and excitation bands are analyzed by the use of energy matrices in order to determine the coupling constant of spin-orbit interaction and the crystal field parameters V₄^(f), V₆^(f) for the 4f orbit.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 69 1 2008 Observation of micropores in hard-carbon using Xe-129 NMR porosimetry 147 152 EN Kazuma Gotoh Takahiro Ueda Hironori Omi Taro Eguchi Mariko Maeda Michihisa Miyahara Nagai Aisaku Hiroyuki Ishida <p>The existence of micropores and the change of surface structure in pitch-based hard-carbon in xenon atmosphere were demonstrated using Xe-129 NMR. For high-pressure (4.0 MPa) Xe-129 NMR measurements, the hard-carbon samples in Xe gas showed three peaks at 27, 34 and 210 ppm. The last was attributed to the xenon in micropores (<1 nm) in hard-carbon particles. The NMR spectrum of a sample evacuated at 773 K and exposed to 0.1 MPa Xe gas at 773 K for 24 h showed two peaks at 29 and 128 ppm, which were attributed, respectively, to the xenon atoms adsorbed in the large pores (probably mesopores) and micropores of hard-carbon. With increasing annealing time in Xe gas at 773 K, both peaks shifted and merged into one peak at 50 ppm. The diffusion of adsorbed xenon atoms is very slow, probably because the transfer of molecules or atoms among micropores in hard-carbon does not occur readily. Many micropores are isolated from the outer surface. For that reason, xenon atoms are thought to be adsorbed only by micropores near the surface, which are easily accessible from the surrounding space.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 95 23 2005 Opacity effect on extreme ultraviolet radiation from laser-produced tin plasmas EN Shinsuke Fujioka Hiroaki Nishimura Katsunobu Nishihara Akira Sasaki Atsushi Sunahara Tomoharu Okuno Nobuyoshi Ueda Tsuyoshi Ando Yezheng Tao Yoshinori Shimada Kazuhisa Hashimoto Michiteru Yamaura Keisuke Shigemori Mitsuo Nakai Keiji Nagai Takayoshi Norimatsu Takeshi Nishikawa Noriaki Miyanaga Yasukazu Izawa Kunioki Mima <p>Opacity effects on extreme ultraviolet (EUV) emission from laser-produced tin (Sn) plasma have been experimentally investigated. An absorption spectrum of a uniform Sn plasma generated by thermal x rays has been measured in the EUV range (9-19 nm wavelength) for the first time. Experimental results indicate that control of the optical depth of the laser-produced Sn plasma is essential for obtaining high conversion to 13.5 nm-wavelength EUV radiation; 1.8% of the conversion efficiency was attained with the use of 2.2 ns laser pulses.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 70 23 1997 Scanning tunneling microscopy/spectroscopy studies of two isomers of Ce@C82 on Si(111)-(7×7&#65289;surfaces EN Satoshi Fujiki Yoshihiro Kubozono Yoshie Rikiishi Tsuneo Urisu <p>Scanning tunneling microscopy images for two isomers of Ce@C-82 were observed on Si(111)-(7x7) at 295 K. The Ce@C-82 molecules in the first layer were bound to the Si surfaces, and the motions were frozen even at 295 K. The multilayer of the Ce@C-82 isomer I (Ce@C-82-I) produced a close-packed structure in the surface layer by annealing the Si substrate at 473 K. The distance between the nearest-neighboring molecules was 1.15(4) nm whose value was consistent with that, 1.12 nm, estimated from x-ray diffraction of the Ce@C-82-I crystals. This implies that the close-packed structure is dominated by van der Waals forces, as in crystals of Ce@C-82-I. The internal structure of Ce@C-82-I was observed in the first layer due to a freeze of molecular motion caused by strong interactions between the molecule and the Si adatoms in the surface. Scanning tunneling spectroscopy revealed that the energy gaps for Ce@C-82-I and -II in the first layer opened to gap energies, E-g of 0.7 and 1.0 eV, respectively. This fact suggests that these molecules are semiconductors with smaller value of E-g than those for C-60 and C-70.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2008 Effect of glucose and pyruvate on nuclear and cytoplasmic maturation of porcine oocytes in a chemically defined medium EN Hiroaki Funahashi Takayuki Koike Ryoko Sakai <p>The objective was to examine potential roles of glucose and pyruvate in nuclear and cytoplasmic maturation of porcine oocytes. Oocyte-cumulus complexes (OCC), derived from 3 to 6 mm follicles, were cultured in a chemically defined medium (pyruvate-free mNCSU37-PVA), with or without 5.55 mM glucose, during in vitro maturation (IVM); in the absence of glucose, germinal vesicle breakdown (GVBD) and nuclear maturation were prevented (P &#60; 0.05). Subsequently, OCC were cultured for IVM in glucose-containing mNCSU37-PVA supplemented with 6-amononicotinamide (6-AN) and diphenyleneiodonium (DPI), inhibitors of the pentose phosphate pathway (PPP); both compounds (&#62;= 10 mu M 6-AN and &#62;= 10 nM DPI) inhibited resumption of meiosis (P &#60; 0.05). Supplementation of glucose-free maturation medium with increasing concentrations of pyruvate induced resumption of meiosis and increased the incidence of oocytes reaching metaphase-II in a concentration-dependent manner (P &#60; 0.05). More mature oocytes were obtained in the presence of pyruvate + glucose (P &#60; 0.05). After culture to allow maturation, glutathione content was higher in oocytes cultured in the presence of pyruvate alone than in those cultured in glucose alone; inclusion of 6-AN abolished responses to pyruvate (P &#60; 0.05). In conclusion, both glucose and pyruvate played a critical role in the release of porcine oocytes from arrest at the GV-I stage, probably through the PPP, whereas supplementation with pyruvate improved cytoplasmic maturation, as determined by oocyte glutathione content.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 68 2 2005 Determination of trace amounts of bromide by flow injection/stopped-flow detection technique using kinetic-spectrophotometric method 274 280 EN K. Uraisin D. Nacapricha S. Lapanantnoppakhun K. Grudpan Shoji Motomizu <p>A simple, sensitive and selective method for the determination of bromide in seawater by using a flow injection/stopped-flow detection technique was examined. The detection system was developed for a new kinetic-spectrophotometric determination of bromide in the presence of chloride matrix without any extraction and/or separation. The detection was based on the kinetic effect of bromide on the oxidation of methylene blue (MB) with hydrogen peroxide in a strongly acidic solution. Large amounts of chloride could enhance the sensitivity of the method as an activator. The decolorisation of the blue color of MB was used for the spectrophotometric determination of bromide at 746 nm. A stopped-flow approach was used to improve the sensitivity of the measurement and provide good linearity of the calibration over the range of 0-3.2 p,g ml(-1) of bromide. The relative standard deviation was 0.74% for the determination of 2.4 jig ml(-1) bromide (n=5). The detection limit (3 sigma) was 0.1 mu g ml(-1) with a sampling frequency of 12 h(-1). The influence of potential interfering ions was studied. The proposed method was applied to the determination of bromide in seawater samples and provided satisfactory results. </p> No potential conflict of interest relevant to this article was reported.

The Japan Society for Analytical Chemistry Acta Medica Okayama 0910-6340 21 3 2005 Trace and ultratrace analysis of purified water samples and hydrogen peroxide solutions for phosphorus by flow-injection method 263 268 EN Zenhai Li Mitsuko Oshima Akhmad Sabarudin Shoji Motomizu <p>A highly sensitive fluorescence-quenching method for the determination of phosphorus based on the formation of an ion associate between molybdophosphate and Rhodamine B (RB) was developed. A simple flow-injection system coupled with a fluorescence detector was used to measure the fluorescence intensity at 560 nm and 580 nm as an excitation and an emission wavelength, respectively. The calibration graph for phosphorus showed a good linearity in the range of (0-1) x 10(-7) M (1 M = 1 mol L-1), and a detection limit of I x 10(-9) M (SIN = 3). The proposed method was successfully applied to the determination of ultratrace amounts of phosphorus in ultrapurified and purified water samples, and to the determination of trace amounts of phosphorus in commercially-available hydrogen peroxide solutions with satisfactory results. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 60 6 2003 Novel flow injection-fluorometric method for the determination of trace silicate and its application to ultrapurified water analysis 1277 1285 EN Akhmad Sabarudin Mitsuko Oshima Naoe Ishii Shoji Motomizu <p>A highly sensitive fluorescence quenching method for the determination of silicate based on the formation of an ion associate between molybdosilicate and Rhodamine B (RB) in nitric acid medium was developed. A flow injection system coupled with a fluorescence detector was used for the measurement of fluorescence intensity at 560 and 580 nm as excitation and emission wavelengths, respectively. The calibration graph for Si showed a linear range of 0.1-5 ng cm(-3) with correlation coefficient of 0.9999, and the detection limit of 0.06 ng cm(-3). The proposed method was successfully applied to the determination of silicate in ultrapurified water with satisfactory results.</p> No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0003-2670 532 1 2005 Slope comparison method (SCM) for the determination of trace amounts of silicate in ultrapurified water 27 35 EN Akhmad Sabarudin Mitsuko Oshima Shoji Motomizu A sensitive analytical method for the determination of trace amounts of silicate in ultrapurified water was developed. The method is based on the formation of an ion associate of molybdosilicate with malachite green (MG) and the collection of the ion associate on a tiny membrane filter (diameter: 5 mm, and effective filtering diameter: 1 mm). The ion associate formed on the membrane filter is dissolved together with the membrane filter in 1 ml of methyl cellosolve (MC) and the absorbance of MC solution is measured at 627 nm by a flow injection-spectrophotometric detection technique. In this method, silicate in the original sample (ultrapurified water) is concentrated as the ion associate into a small volume of MC to get high sensitivity. As sample concentration takes place, the small amounts of silicate contained in the reagents used also become concentrated as the ion associate into MC. The original sample volumes are varied and evaporated to an identical volume. Therefore, the reagent added is fixed to the same volume. The absorbance increase linearly with increase in the original sample volume will be due only to silicate in the original samples (ultrapurified water). The resulting slopes obtained by varying the sample volumes are compared with the slope of the calibration graph, and thus named the slope comparison method (SCM). The SCM facilitates a more sensitive and accurate evaluation of silicate concentration in the samples than either common calibration method (CCM) or standard addition method (SAM) because it compensates for the influence of trace amounts of silicate contained in chemicals, reagent solution and solvent used. The calibration graph was constructed from 0 to 0.25 ng ml(-1) of Si and the detection limit was 10 pg ml(-1) (ppt) when 30 ml of samples was used. The standard deviation and relative standard deviation from six measurements of the reagent blanks were 0.0012 and 3.5%, respectively. No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0003-2670 612 2 2008 Development of novel detection reagent for simple and sensitive determination of trace amounts of formaldehyde and its application to flow injection spectrophotometric analysis 165 172 EN Qiong Li Piyanete Sritharathikhum Mitsuko Oshima Shoji Motomizu In this paper, a novel detection reagent for formaldehyde determination is proposed, and is applied to a simple and highly sensitive flow injection method for the spectrophotometric determination of formaldehyde. The method is based on the reaction of formaldehyde with methyl acetoacetate in the presence of ammonia. The increase in the absorbance of the reaction product was measured at 375 nm. An inexpensive light emitting diode (LED)based UV detector (375 nm) was, for the first time, used. Under the optimized experimental conditions, formaldehyde in an aqueous solution was determined over the concentration range from 0.25 to 20.0 x 10(-6) M with a liner calibration graph; the limit of detection (LOD) of 5 x 10(-8) M (1.5 mu g L-1) was possible. The relative standard deviation of 12 replicate measurements of 5 X 10(-6) M formaldehyde was 1.2%. Maximum sampling throughput was about 21 samples/h. The effect of potential interferences such as metals, organic compounds and other aldehyde was also examined. The analytical performance for formaldehyde determination was compared with those obtained by the conventional acetylacetone method, which uses visible absorption spectrophotometry. Finally, the proposed method was successfully applied to the determination of formaldehyde in natural water samples. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 23 4 2007 Development of novel reagent for Hantzsch reaction for the determination of formaldehyde by spectrophotometry and fluorometry 413 417 EN Qiong Li Piyanete Sritharathikhun Shoji Motomizu <p>A novel reagent, acetoacetanilide (AAA), was introduced to the determination of formaldehyde based on Hantzsch reaction. A simple and highly sensitive fluorometric method was achieved by using AAA. The main advantages in the use of this reagent are: the reaction is carried out at room temperature without any heating system, the cyclization product based on Hantzsch reaction is soluble in water, and the product can be detected by spectrophotometry and fluorometry. The maximum absorption wavelength of the product occurs at 368 nm, and the maximum excitation and emission wavelengths are found at 370 and 470 nm, respectively. Several important experimental variables of the procedures were examined; particularly, the reaction temperature, reaction time, concentrations of reagents, and pH of the reagent solution were optimized for improving the detecting sensitivity. The calibration graph was linear in the range of 1 x 10(-7) -1 X 10(-6) M or much higher concentrations. The limit of detection (LOD), based on three times of the standard deviation of the reagent blank, was 2.0 x 10(-8) M. The proposed method was applied to the determination of formaldehyde in environmental water samples. Many foreign species commonly existing in water samples did not interfere with the determination of formaldehyde in the proposed method.</p> No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0003-2670 481 2 2003 Ultratrace determination of phosphorus in ultrapurified water by a slope comparison method 311 319 EN Akhmad Sabarudin Mitsuko Oshima Shoji Motomizu The analytical method for the determination of phosphorus in ultrapurified water was developed. Ultrapurified water was evaporated to concentrate phosphorus and the final sample volume for analysis was 10 ml. In 0.55 mol 1(-1) HCl, orthophosphate forms molybdophosphate, and then the molybdophosphate forms ion associate with Malachite Green (MG), which can be collected on a tiny membrane filter (diameter: 5 mm, and effective filtering diameter: 2 mm). After the ion associate on the membrane filter is dissolved together with the membrane filter in I ml of methyl cellosolve (MC), the absorbance of MC solution is measured at 627 nm by a flow injection-spectrophotometric detection technique. When 10 ml of the sample solution was used for the procedures and absorbance measurement, the calibration graph is linear up to about 500 ng 1(-1) of phosphorus and the detection limit was 8 ng 1(-1) (S/N = 3). For the determination of phosphorus in an ultrapurified water, 10-40 ml of sample solutions were transferred into poly(tetrafluoroethylene) (PTFE) beaker and evaporated to 5 ml or to dryness. To them, 0.003 mol 1(-1) HCl was added to get 10 ml of final solution, which was used as sample. Phosphate is determined by comparing the slope of the varied sample volume after evaporation/concentration with a slope of the standard calibration graph (a slope comparison method: SCM). The SCM enables to evaluate the concentration of phosphate in ultrapurified waters more sensitively and accurately. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 23 5 2007 Simultaneous measurements of K+ and calcein release from liposomes and the determination of pore size formed in a membrane 517 522 EN Takashi Katsu Tomonori Imamura Keiko Komagoe Kazufumi Masuda Tohru Mizushima <p>The changes induced by biologically active substances in the permeability to K+ and calcein of liposomes composed of egg phosphatidylcholine and cholesterol were measured simultaneously in order to rapidly screen the sizes of pores formed in a membrane, using different sized markers. The substances examined in the present study were classified into three types based on differences in the rates at which K+ and calcein were released. The first type released only K+, and included gramicidin A. The second type predominantly released K+, preceding the release of calcein, and included amphotericin B and nystatin. The third type, including antimicrobial peptides, such as gramicidin S, alamethicin, and melittin, and several membrane-active drugs, like celecoxib (non-steroidal anti-inflammatory drug), 1-dodecylazacycloheptan-2-one (named azone; skin permeation enhancer), and chlorpromazine (tranquilizer), caused the release of K+ and calcein simultaneously. Thus, the sizes of pores formed in a liposomal membrane increased in the following order: types one, two, and three. We determined the size more precisely by conducting an osmotic protection experiment, measuring the release of calcein in the presence of osmotic protectants of different sizes. The radii of pores formed by the second type, amphotericin B and nystatin, were 0.36 - 0.46 nm, while the radii of pores formed by the third type were much larger, 0.63 - 0.67 nm or more. The permeability changes induced by substances of the third type are discussed in connection with a transient pore formed in a lipid packing mismatch taking place during the phase transition of dipalmitoylphosphatidylcholine liposomes.</p> No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0006-8993 961 2 2003 A Periaxonal Net in the Zebrafish Central Nervous System 179 189 EN Hiroshi Nakayasu Noriko Arata We produced a monoclonal antibody, named A20, which specifically recognizes a 35 kDa protein and stains myelinated axons in zebrafish brain. The A20 antigen is located at the outside of the myelin layer of large axons, and comprises a fine meshwork composed of thin unit fibers about 1–2 μm in length and about 100–200 nm in thickness. The unit fibers form pentagonal and hexagonal structures, which further polymerize into an envelope structure on the axons. The A20 monoclonal antibody did not stain neuronal cell bodies nor synapses. Instead, the distribution of the A20 antigen was along axons, practically coincident with the distribution of myelin basic protein. The monoclonal antibody stained only axons in the central nervous system (CNS), and not the extracellular matrix surrounding Schwann cells. These results suggest that this antigenic meshwork (which we call the periaxonal net) is synthesized by oligodendrocytes. During the development of the zebrafish brain, the periaxonal net appeared after the formation of myelin on the axons. The periaxonal net developed first at the brain stem, then gradually appeared at the caudal end of the spinal cord. The thickness of the periaxonal net around the Mauthner axon changed during development. Although the thickness of the Mauthner axon continues to grow throughout life, the thickness of periaxonal net stopped growing at 6 months after fertilization. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 207 4 2004 Purification and cDNA cloning of the ovigerous-hair stripping substance (OHSS) contained in the hatch water of an estuarine crab Sesarma haematocheir 621 632 EN Oleg Gusev Hideki Ikeda Tetsushi Okochi Jae Min Lee Masatsugu Hatakeyama Chiyoko Kobayashi Kiyokazu Agata Hidenori Yamada Masayuki Saigusa <p>The egg attachment system of an estuarine crab Sesarma haematocheir is formed on the maternal ovigerous hairs just after egg laying, and slips off these hairs just after hatching. The stripping is caused by an active factor that we call OHSS (ovigerous-hair stripping substance), which is released by the embryo upon hatching. OHSS was purified, and its active form had a molecular mass of 25·kDa. The cDNA of OHSS cloned from an embryonic cDNA library was 1759·bp long, encoding 492 amino acids in a single open reading frame (ORF). The C-terminal part of the predicted protein was composed of a trypsin-like serine protease domain, with homology to counterparts in other animals of 33–38%. The predicted protein (54.7·kDa) secreted as a zymogen may be cleaved post-translationally, separating the Cterminal from the N-terminal region. The OHSS gene was expressed in the embryo at least 2 weeks before hatching. Expression was also detected in the zoea larva 1 day after hatching and in the brain of the female. However, it was not detected in the muscle, hepatopancreas or ovigerous seta of the female. Ultrastructural analysis indicated that the material investing maternal ovigerous hair, i.e. the outermost layer (E1) of the egg case, is attached at the special sites (attachment sites) arranged at intervals of 130–160·nm on the hair. It is suggested that OHSS acts specifically at these sites, lysing the bond with the coat, thus disposing of the embryo attachment system. This enables the female to prepare the next clutch of embryos without ecdysis.</p> No potential conflict of interest relevant to this article was reported.

Elsevier Ireland Ltd. Acta Medica Okayama 0304-3835 224 2 2005 Significant growth suppression of synovial sarcomas by the histone deacetylase inhibitor FK228 in vitro and in vivo 311 319 EN Tatsuo Ito Mamoru Ouchida Yuki Morimoto Aki Yoshida Yoshimi Jitsumori Toshifumi Ozaki Hiroshi Sonobe Hajime Inoue Kenji Shimizu About 97% of synovial sarcomas harbor the SYT-SSX fusion gene by chromosomal translocation. We found that the histone deacetylase (HDAC) inhibitor FK228 significantly suppressed the growth of synovial sarcoma cells as compared with that of osteosarcoma. The 50% growth inhibition IC50 value we obtained for FK228 was 0.02-0.2 nM, and it indicates that its suppression effect on synovial sarcoma cells is the highest of any of the HDAC inhibitors yet reported. It was not likely that the growth suppression of FK228 depends on the doubling time of these cells. Introduction of SYT-SSX cDNA into HEK293 cells enhanced the sensitivity of the cells for FK228. Immunostaining of the FK228-treated cells using an anti-acetyl-histone H3 antibody showed that FK228 inhibits deacetylation of histone. In a mice assay, the growth of synovial sarcoma cells was markedly inhibited by FK228 treatment, and the invasion of tumors into surrounding tissues was suppressed. These results suggest that FK228 may be useful in developing therapeutic strategies to treat synovial sarcoma. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 162 1 2004 Separation into polar and hydrogen-bonding factors of the effects of alcohols on the emission spectrum of 4-phenyl-1-N,N-dimethylaminobutane in THF 73 79 EN Xie Guobin Mayuko Nakamura Yoshimi Sueishi Shunzo Yamamoto <p>The effects of the additions of protic and aprotic polar solvents on the emission spectrum of 4-phenyl-1-N,N-dimethylaminobutane (PDAB) in THF have been studied under conditions of steady-state illumination. The fluorescence spectrum of PDAB in THF was reported to consist of three component bands (band A at 285 nm (fluorescence of the phenyl group), band B at 343 nm (fluorescence of the amino group) and band C at 385 nm (emission from an intramolecular exciplex)). The intensities of bands B and C decreased with increasing solvent polarity. They also decreased owing to the hydrogen-bonding interaction between the amino group in PDAB and protic solvents, but in this case the intensity of band A was found to increase. Acetonitrile has only a polar effect and trichloroacetic acid only a hydrogen-bonding (or protonation) effect, while alcohols have both effects. The equilibrium constants for the formation of intermolecular hydrogen-bonded complexes of the amino group with alcohols were estimated from the intensity change of band A. The hydrogen-bonding and polar effects of alcohols on the intensities of bands B and C could be separately evaluated. The decrease in the intensities of bands B and C with increasing solvent polarity in THF-AN and THF-alcohol mixtures is considered to be caused by the conversion of the exciplex to an ion-pair enhanced by the increase in solvent polarity.</p> No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 162 2-3 2004 Effects of alcohols on emission spectra of toluene-triethylamine mixtures in THF: separation into polar and hydrogen-bonding interactions 449 456 EN Guobin Xie Yoshimi Sueishi Shunzo Yamamoto <p>The emission spectra of toluene (T)–triethylamine (TEA) systems were measured under conditions of steady-state illumination in some protic and aprotic solvent–THF mixtures. The fluorescence spectrum of the T–TEA system in THF could be separated into three component bands (band A at 279 nm (fluorescence of T), band B at 336 nm (fluorescence of TEA) and band C at 373 nm (emission from an intermolecular exciplex)). The intensities of bands B and C decreased with increasing solvent polarity. The decrease in the intensities of bands B and C is considered to be caused by the enhanced conversion of the exciplex to an ion-pair with increase in solvent polarity. The intensities of bands B and C also decreased owing to the hydrogen-bonding interaction between TEA and protic solvents, but in this case the intensity of band A increased. Acetonitrile only has a polar effect and trichloroacetic acid only has a hydrogen-bonding (or protonation) effect, while alcohols have both effects. The equilibrium constants for the formation of intermolecular hydrogen-bonded complexes of TEA with alcohols were estimated from the changes in the intensity of band A. The hydrogen-bonding and polar effects of alcohols on the intensities of bands B and C could be evaluated separately. The ratio of the hydrogen-bonding effect to the polar effect of alcohols was observed to increase with increasing vol.% of alcohol.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 37 3 1983 Characteristics of muscarinic acetylcholine receptors in rat brain. 179 191 EN Itaru Nukina Article 10.18926/AMO/32433 <p>Characteristics of muscarinic acetylcholine (ACh) receptors were studied in the rat central nervous system (CNS) using 3H-quinuclidinyl benzilate (QNB), an antagonist of muscarinic ACh receptors. Scatchard analysis indicated that the rat CNS had a single 3H-QNB binding site with an apparent dissociation constant (Kd) of 5.0 X 10(-10) M. Li+, Zn++ and Cu++ had strong effects on 3H-QNB binding which indicates that these metal ions might play important roles at muscarinic ACh receptor sites in the brain. Since antidepressants and antischizophrenic drugs displaced the binding of 3H-QNB, the anticholinergic effects of these drugs need to be taken into account when they are applied clinically. The muscarinic ACh receptor was successfully solubilized with lysophosphatidylcholine. By gel chromatography, with a Sepharose 6B column, the solubilized muscarinic ACh receptor molecule eluted at the fraction corresponding to a Stokes' radius of 6.1 nm. With the use of sucrose-density-gradient centrifugation, the molecular weight of the solubilized muscarinic ACh receptor was determined to be about 90,000 daltons. The regional distribution of 3H-QNB binding in rat brain was examined, and the highest level of 3H-QNB binding was found to be in the striatum followed by cerebral cortex and hippocampus, indicating that muscarinic ACh mechanisms affect CNS function mainly through these areas.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 37 2 1983 Molecular sieve in bovine descemets membrane as revealed by negative staining. 155 157 EN Hirofumi Makino Hiroyuki Toyofuku Yasuaki Mino Michio Takaoka Zensuke Ota Brief Note 10.18926/AMO/32417 <p>Descemet's membrane was isolated from the corneas of cows and observed by electron microscopy after negative staining with 1% phosphotungstic acid solution, pH 7.2. Ultrastructurally, bovine Descement's membrane had a very regular hexagonal pattern. Nodes were connected to the six others around each of them by thin filaments to form a hexagon. The distance between the nodes was approximately 120 nm, the diameter of the nodes approximately 30 nm, and the width of the connecting filaments approximately 10 nm. Bovine Descement's membrane was a molecular sieve composed of nodes and filaments substantiating our molecular sieve theory of basement membranes.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 37 2 1983 Modulation of optical density by sulfhydryl reagents in microbiuret method: a modified method for protein determination in the presence of sulfhydryl reagents. 125 129 EN Hideki Matsui Tomohiro Kurosaki Masaaki Tokuda Osamu Hatase Article 10.18926/AMO/32414 <p>2-Mercaptoethanol increases the optical density of assay solutions at wavelengths between 280 to 400 nm, and therefore interferes with the measurement of protein concentration by the microbiuret method. Protein concentration can be determined in the presence of 2-mercaptoethanol up to 6 mM by modification of the method as follows: after the precipitation of protein by trichloroacetic acid in the presence of deoxycholate, the precipitate is resolubilized with NaOH solution. Dithiothreitol interfered with the protein determinations could by made in the presence of 4 mM of dithiothreitol with the modified microbiuret method. This modified method is time-saving and more reliable than other methods for protein determination, such as Lowry's method, in the presence of sulfhydryl reagents.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 37 2 1983 Characteristics of gamma-aminobutyric acid (GABA) receptors in the rat central nervous system. 93 106 EN Hiroo Kuroda Article 10.18926/AMO/32411 <p>Characteristics of gamma-aminobutyric acid (GABA) were investigated in the rat central nervous system by radioreceptor assay (RRA). Scatchard analysis revealed that the rat brain had two distinct GABA binding sites with an apparent dissociation constant (Kd) of 11.7 nM and 34.7 nM. The highest level of specific [3H]-GABA binding was found in the rat cerebellum. Imidazole acetic acid, a potent GABA agonist, was effective in displacing [3H]-GABA binding but beta-alanine was slightly effective in inhibiting [3H]-GABA binding. Muscimol, the most potent GABA agonist, has been used as a ligand to characterize the postsynaptic GABA receptors. However, the maximal binding capacity (Bmax) of muscimol-RRA was about 3 times larger than that of GABA-RRA, suggesting that muscimol might label not only GABA receptors but other unknown receptors as well. An endogenous inhibitor of GABA receptor binding was purified from the P2 fraction of rat brain with 0.05% Triton X-100. The endogenous inhibitor was competitive with GABA on GABA binding sites. The inhibition by the endogenous inhibitor of GABA receptor binding was blocked by the allosteric effect of diazepam. In the presence of diazepam, [3H]-GABA binding with the endogenous inhibitor was larger than that with GABA, whereas there was no difference in the absence of diazepam. This indicated that the endogenous inhibitor was not GABA itself. The molecular weight of the endogenous inhibitor was estimate by gel filtration to be less than 3,000 daltons.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 54 4 2000 Inhibitory actions of hydroxocobalamin, cyanocobalamin, and folic acid on the ultraviolet light-induced relaxation of the frog upper oesophageal strip. 147 152 EN Peyman Ucar Cemil Gocmen Ata Secilmis Yusuf Karatas Ergin Singirik Serpil Onder Atilla Dikmen Firuz Baysal Article 10.18926/AMO/32273 <p>The applications of ultraviolet (UV) light (336 nm) on the upper oesophageal strips of frog elicited relaxant responses in the presence of NaNO2 (50 microM). The tissues were mounted under the tension 0.5 g in an organ bath containing Ringer solution, maintained at 25 degrees C and gassed with 100% O2. The responses were recorded on a kymograph via an isotonic lever. Antimegaloblastic agents, including hydroxocobalamin (1, 10, and 100 microM), cyanocobalamin (1, 10, 25, and 100 microM), and folic acid (1, 10, 50, 100, and 200 microM), significantly attenuated the relaxation response to UV light. Folinic acid (1, 10, 25, and 100 microM), however, enhanced the relaxation. Pyrogallol (50 microM), hydroquinone (50 microM), and diethyldithiocarbamic acid (8 mM) were found ineffective for attenuation, though FeSO4 (200, 400, and 500 microM) and hemoglobin (50 microM), respectively, exerted significant inhibition. L-arginine methylester (500 microM) did not impair UV-induced relaxation. Based on these results, we concluded that a mechanism involving undefined action(s) of antimegaloblastic drugs may cause alterations in the UV light-induced relaxation of the tissue used.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 45 1 1991 Physicochemical and immunological properties of the hepatitis B surface antigen containing the preS2 9 amino acid sequence produced by a recombinant yeast. 11 19 EN Takeo Konobe Nobuyoshi Ishikawa Hideo Gohda Konosuke Fukai Akinobu Okabe Article 10.18926/AMO/32216 <p>The hepatitis B virus surface antigen containing the preS2 nine amino acid sequence produced by a recombinant Saccharomyces cerevisiae (yHBsAg) was purified and its physicochemical properties were determined. Ultrastructurally, the yHBsAg was found to be a homogeneous spherical particle with a diameter of 24 +/- 4 nm. The homogeneity of the yHBsAg particles was also demonstrated by analyses of their buoyant density and isoelectric point. They consisted of protein (53%), lipid (36%) and carbohydrate (11%), and the alpha-helix content was estimated to be 32%, differing from the reported values for human plasma-derived HBsAg (hHBsAg). Immunodiffusion analysis showed that the antigenic specificity of yHBsAg was identical to that of hHBsAg. Immunization of mice demonstrated that the immunogenicity of the yHBsAg was significantly higher than that of hHBsAg. </p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 33 5 1979 The mechanism of the release of hepatic enzymes in various liver diseases. 1. Alterations in cytoplasmic and mitochondrial enzyme activities in serum. 287 304 EN Shu Miyake Article 10.18926/AMO/32061 <p>Serum glutamic oxaloacetic transaminase (GOT), mitochondrial GOT (GOTm), glutamic-pyruvic transaminase (GPT) and glutamate dehydrogenase activities were determined in 43 healthy controls and in 280 cases of liver diseases. A simplified column chromatographic method coupled with UV assay was employed for separation of GOTm. The activity was measured by following decrease in abosrbance of NADH at 340 nm. The lowest activity of GOTm determined with a coefficient of variation below 10% was 6 mIU/ml. High GOTm activities were found in acute hepatitis (acute stage), subacute hepatitis and primary biliary cirrhosis and were generally associated with high total GOT (GOTt) activities. The activity ratio of GOTm/GOTt varied depending on the stage and severity of liver diseases. The GOTm/GOTt ratio was decreased in acute, fulminant and subacute hepatitides. No significant reduction in the ratio was found in bile duct obstruction, alcoholic liver injury or metastatic liver cancer. Although relatively high GOTm/GOTt ratios were found in some patients with severe hepatic injury, they had no definite association with poor prognosis. These results indicate that the marked elevation in GOTt over GPT in advanced chronic hepatitis, liver cirrhosis and primary hepatoma was mainly due to preferential leakage of cytoplasmic GOT (GOTs).</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 33 4 1979 The mechanism of the release of hepatic enzymes in various liver diseases. 1. Alterations in cytoplasmic and mitochondrial enzyme activities in serum. 287 304 EN Shu Miyake Article 10.18926/AMO/32034 <p>Serum glutamic oxaloacetic transaminase (GOT), mitochondrial GOT (GOTm), glutamic-pyruvic transaminase (GPT) and glutamate dehydrogenase activities were determined in 43 healthy controls and in 280 cases of liver diseases. A simplified column chromatographic method coupled with UV assay was employed for separation of GOTm. The activity was measured by following decrease in abosrbance of NADH at 340 nm. The lowest activity of GOTm determined with a coefficient of variation below 10% was 6 mIU/ml. High GOTm activities were found in acute hepatitis (acute stage), subacute hepatitis and primary biliary cirrhosis and were generally associated with high total GOT (GOTt) activities. The activity ratio of GOTm/GOTt varied depending on the stage and severity of liver diseases. The GOTm/GOTt ratio was decreased in acute, fulminant and subacute hepatitides. No significant reduction in the ratio was found in bile duct obstruction, alcoholic liver injury or metastatic liver cancer. Although relatively high GOTm/GOTt ratios were found in some patients with severe hepatic injury, they had no definite association with poor prognosis. These results indicate that the marked elevation in GOTt over GPT in advanced chronic hepatitis, liver cirrhosis and primary hepatoma was mainly due to preferential leakage of cytoplasmic GOT (GOTs).</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 55 5 2001 Influence of tumor thickness and vascularity on survival in cutaneous melanoma. 289 293 EN Gordana Zamolo Franjo Gruber Leo Cabrijan Vladimir Micovic Zlatko Iternicka Nives Jonjic Article 10.18926/AMO/32021 <p>Eighty-eight cases of cutaneous melanoma (CM) were analyzed in order to elucidate the relationship between thickness, angiogenesis, and prognosis. The thickness of the tumor was measured according to the Breslow method, and the microvessels were identified by an immunohistochemical study using anti-factor VIII monoclonal antibody on specimens from 40 patients with superficial spreading melanoma (SSM), and 48 with nodular type (NM). Microvessels were counted in the area of highest density. The overall survival and disease-free period were analyzed retrospectively. The proportion of patients with thicker CMs (&#62; 1.5 mm) increased with age in both sexes. Mean vascular count was statistically significant different only between thinner and thicker tumors in the SSM group (P &#60; 0.05). Prognosis was correlated with the thickness of CM (P = 0.0002), mean vascular count alone (P = 0.004), mean vascular count in association with CM thickness less than 1.5 mm (P = 0.0005), and with mean vascular count in NM (P = 0.02). These findings suggest that increasing microvessel density indicates a worsening prognosis.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 40 1 1986 Serotonin-containing nerve fibers in the rat spinal cord: electron microscopic immunohistochemistry. 1 10 EN Kiminao Mizukawa Nagayasu Otsuka Toshiaki Hattori Article 10.18926/AMO/31945 <p>The ultrastructure of the serotonin (5HT) system in the spinal cord of rats was studied by an immunohistochemical peroxidase-antiperoxidase (PAP) method. Under the light microscope, 5HT immunoreactive staining was observed as brown-colored dots in the anterior horn, lateral horn, posterior horn and pericentral canal region. These positively staining dots were probably indicative of 5HT immunoreactive varicosities and nerve terminals. At the ultrastructural level, 5HT immunoreactive nerve fibers appeared as darkly stained varicosities with PAP positive large electron dense vesicles (80-100 nm), as well as small clear vesicles (30-40 nm) finely coated with PAP immunoreactive products. In the anterior horn, some of the 5HT immunoreactive structures were clearly nerve terminals forming asymmetric synaptic contact with soma or dendrites of the anterior horn cells. In the lateral horn, posterior horn and pericentral canal region, however, only 5HT positive varicosities were detected.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 40 4 1986 Tubular structures in the hepatic nuclei of a patient with delta agent. 225 228 EN Gotaro Yamada Koji Manabe Hideo Nagashima Article 10.18926/AMO/31906 <p>Liver biopsy specimens obtained from a 31-year-old female with delta-positive hepatitis were studied by routine electron microscopy. In several nuclei of hepatocytes, there were filamentous or microtubular structures 15 to 20 nm in diameter, in the vicinity of which, round particles, probably cross sections of tubular ones, were seen. In these nuclei, irregular granules approximately 20 to 30 nm in diameter were also found in clusters. However, cores of Dane particles were not found in such hepatocytes. These intranuclear microtubular structures may be associated with delta agent.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 41 2 1987 Electron microscopic study of mossy fiber endings of the hippocampal formation in El mice. 81 84 EN Toshikazu Hochi Hiroki Watanabe Kiminao Mizukawa Nagayasu Otsuka Article 10.18926/AMO/31749 <p>The effects of seizure activity on the mossy fiber endings of El mice were studied by electron microscopy. During epileptic seizures of El mice, the number of clear round vesicles (50 nm) in the mossy fiber endings of the hippocampal formation decreased, while the number of large densecore vesicles (100 nm) increased. In these endings, the large dense-core vesicles were scattered during the resting state, but after seizure activity they tended to accumulate together and attach to the presynaptic membrane. Omega-shaped profiles, which seemed to be due to exocytosis of the large dense-core vesicles, were seen in the presynaptic membrane.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 41 6 1987 Determination of Sialic Acids by Acidic Ninhydrin Reaction 237 241 EN Kenzabroh Yao Toshihiko Ubuka Article 10.18926/AMO/31741 <p>A new acidic ninhydrin method for determining free sialic acids is described. The method is based on the reaction of sialic acids with Gaitonde's acid ninhydrin reagent 2 which yields a stable color with an absorption maximum at 470 nm. The standard curve is linear in the range of 5 to 500 nmol of N-acetylneuraminic acid per 0.9 ml of reaction mixture. The reaction was specific only for sialic acids among the various sugars and sugar derivatives examined. Some interference of this method by cysteine, cystine and tryptophan was noted, although their absorption maxima differed from that of sialic acids. The interference by these amino acids was eliminated with the use of a small column of cation-exchange resin. The acidic ninhydrin method provides a simple and rapid method for the determination of free sialic acids in biological materials.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 41 4 1987 Fibrillar ultrastructure of the glomerular basement membrane of the rat kidney as revealed by digestive treatment. 183 185 EN Akira Ichiyasu Michio Takaoka Hirofumi Makino Kayo Takahashi Zensuke Ota Article 10.18926/AMO/31729 <p>Rat kidney glomerular basement membrane (GBM) was isolated and digested with alpha-amylase and elastase. Electron microscopy revealed a meshwork structure composed of fibrils 3 nm in width. They appeared to be type IV collagen fibrils. We succeeded in clarifying a significant ultrastructural aspect of the GBM which had been unclear until now. The findings are consistent with our previously proposed GBM molecular sieve theory.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 53 3 1999 Urinary excretion of type I collagen cross-linked N-telopeptides, bone mass and related lifestyle in middle-aged women. 133 140 EN Chie Masatomi Kaori Imai Da-Hong Wang Satoru Ikeda Kazuhisa Taketa Shinji Takata Shohei Kira Article 10.18926/AMO/31637 <p>The relationship between past and present lifestyle and urinary excretion of type I collagen cross-linked N-telopeptides (NTx) was studied in 61 Japanese females aged 34-59, with a view toward using NTx excretion rates as a predictor of future osteoporosis. Bone mineral density (BMD) of the lumbar spine, the speed of sound (SOS) and broadband ultrasound attenuation (BUA) of the os calcis, urinary NTx, serum osteocalcin (BGP) and bone-specific alkaline phosphatase (BAP) were measured. Stiffness index (stiffness) was calculated from SOS and BUA. The subjects were asked whether they took regular exercise in their childhood and teen years (in elementary, junior-high, senior-high school and college), the past (20-40 years of age) and present adulthood. Regular calcium intake, smoking habits, alcohol and other beverage consumption and milk consumption were also covered in the questionnaire. The mean NTx values of premenopausal and postmenopausal group were 22.2 and 56.0 nM bone collagen equivalents (BCE)/mM urinary creatinine (Cr), respectively. The group which did not exercise regularly between the ages of 20 and 40 had a higher mean NTx value (40.9 nMBCE/mMCr) than the group which did exercise regularly (22.7 nMBCE/mMCr). In multiple regression analyses, age, stiffness and exercise in past adulthood could explain 43.5% of the NTx variance. For prevention of bone metabolic increases around menopause, habitual exercise in early adulthood seems to be effective.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 39 3 1985 Application of Rapid and Simul­taneous Measurement of Sex Steroid Hormones to the Monitoring of Gonadotropin Therapy 161 170 EN Koshi Hayata Nobuyoshi Hayashi Kaoru Sekiba Article 10.18926/AMO/31533 <p>A high performance liquid chromatographic (HPLC) method with both electrochemical detection (ECD) and ultraviolet spectrometric detection (UVD) was developed for the rapid and simultaneous measurement of estradiol (E2), estrone (E1), testosterone (T), 17 alpha-hydroxyprogesterone (17-OHP) and progesterone (P) in serum. These hormones were extracted with diethylether, and chromatographed on an octadecyl silane-silica (ODS) column with an eluent of a phosphate buffer solution - acetonitrile mixture (volume ratio 49:51). Estrogens were detected by ECD at +1.0 V vs. Ag/AgCl, and other hormones by UVD at 242 nm. With this method, the simultaneous determination of sex steroid hormones could be performed within approximately two hours with high precision. The hormones of 34 patients (39 menstrual cycles) undergoing human menopausal gonadotropin (HMG)-human chorionic gonadotropin (HCG) therapy were measured. It was concluded that the switch from HMG to HCG should be performed when the E2 level reaches 400 pg/ml for ovulation and 800 pg/ml for pregnancy. The occurrence of ovarian hyperstimulation syndrome (OHSS) can be predicted when the P level rises above 30 ng/ml on the 7th day after the switch. Moreover, conception may be indicated when the P level does not increase from the 7th to 14th day after the switch. In this way, this method proved to be useful for the monitoring of HMG-HCG therapy.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 35 5 1981 Antimycotic action of a diamidine compound, 1, 4-bis-(m, m'-amidinophenoxymethyl)-cyclohexane dilactate, on Candida albicans. 327 341 EN Tomotsu Kanazawa Article 10.18926/AMO/31277 <p>The antimycotic action of 1, 4-bis-(m, m'-amidinophenoxymethyl)-cyclohexane dilactate (MAC), a synthetic diamidine compound, on Candida albicans was studied. The minimum inhibitory concentration (MIC) ranged from 3.31 to 6.25 micrograms/ml against both standard and clinically isolated strains. MAC was fungistatic at MIC and weakly fungicidal at the concentration of 100 micrograms/ml. MAC did not affect the cell wall or cause cell lysis. Intracellular constituents, such as 260 nm and 280 nm absorbing materials, were released from the cells by treatment with MAC indicating that MAC affected membrane permeability. The release of 260 nm absorbing material was inhibited by the presence of Ca2+ and Mg2+. Acidic phospholipids such as cardiolipin and phosphatidylglycerol inhibited the anti-Candida activity of MAC, but sterols and lecithin were not inhibitory, indicating that MAC interacted with acidic phospholipids of the cell membrane. Freeze-fracture electron microscopy showed that MAC caused aggregation of membrane particles and patch formation on the P face, which may indicate that MAC is a membrane disrupting agent. It appeared that MAC affected C. albicans at the cell membrane by interacting with acidic phospholipids and caused disorganization of the membrane structure resulting in the release of intracellular constituents without lysis.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 31 4 1977 An improved direct colorimetric method for the quantitative analysis of urinary hippuric acid as an index of toluene exposure 235 242 EN Masana Ogata Reiko Sugihara Original Article 10.18926/AMO/30813 <p>An improved direct colorimetric method for determining the concentration of urinary hippuric acid as an index of toluene exposure was described. One tenth ml of urine was diluted with 0.4 ml 0.01 M phosphate buffer H 6.9 and mixed with 0.5 ml pyridine. The mixture was layered on 0.2 ml benzenesulfonyl chloride. The reaction was started by mixing for one min with a mechanical shaker. The colored solution was allowed to stand for 30 min, diluted with 5 ml ethanol, and absorbance measured at 410 nm within 30 min after the dilution. The coefficient of variation of this method was 6% and the recovery 103% when urine contains about 0.2-0.5 mg hippuric acid per ml of urine. The concentration was linear up to 2.0 mg per ml hippuric acid in a specimen.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 60 1 2006 In situ preparation of colloidal iron by microwave irradiation for transmission electron microscopy. 59 64 EN Satoru Nakatani Ichiro Naito Ryusuke Momota Noriko Hinenoya Kanji Horiuchi Keiichiro Nishida Aiji Ohtsuka Article 10.18926/AMO/30753 <p>We attempted to prepare colloidal iron within tissues by means of microwave irradiation. Mouse tissue blocks were fixed with a mixture of paraformaldehyde and ferric chloride in a cacodylate buffer, immersed in a cacodylate buffered ferric chloride solution, and irradiated in a microwave processor. Colloidal iron was prepared within tissues or cells, and was observed in the form of electron dense fine granules (1-2 nm in diameter) by transmission electron microscopy. Collagen fibrils in the connective tissue showed colloidal iron deposition at regular periodical intervals. Cells in the splenic tissue showed that fine colloidal granules were deposited on the ribosomes but not on the nuclear chromatin. This finding suggests that ferric ions could not diffuse into the nucleus, which was surrounded by the nuclear envelope. The podocyte processes of the renal glomerulus were stained diffusedly. Though this microwave in situ colloidal iron preparation method has some limitations, it is convenient for use in biomedical specimen preparation in transmission electron microscopy.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 36 5 1982 Molecular sieve in rat glomerular basement membrane as revealed by negative staining. 371 382 EN Hirofumi Makino Article 10.18926/AMO/30692 <p>Human and bovine glomerular basement membranes (GBM) were previously shown to be a three-dimensional molecular sieve composed of pores and strands by negative staining and electron microscopy. In this study, rat GBM were isolated under several different conditions to rule out morphological changes due to isolation procedures. Rat GBM isolated under different conditions all showed the same morphological features as bovine and human GBM. The strands forming the molecular sieve were almost equal in width, measuring approximately 3.1 +/- 0.8 nm. Pores were oval or polygonal. The size of pores varied a little averaging 4.4 +/- 1.0 nm in the long dimension and 3.0 +/- 0.6 nm in the short dimension. The average density of the pores was 16 +/- 2/1,000 nm2. Negative staining demonstrated pores in isolated and unfixed GBM, indicating that the function of GBM is mechanical filtration of macromolecules on the basis of size.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 44 2 1990 Visualization of sialoglycoproteins in polyacrylamide gels by acidic ninhydrin reaction. 65 70 EN Kenzabroh Yao Toshihiko Ubuka Noriyoshi Masuoka Masahiro Kinuta Jun Ohta Kazushi Ishino Article 10.18926/AMO/30439 <p>A new method for staining sialoglycoproteins in polyacrylamide gel after disc electrophoresis is described. The method utilizes the reaction of sialic acids with an acidic ninhydrin reagent which yields a stable color with an absorption maximum at 470 nm. After electrophoresis, the polyacrylamide gel is placed in a test tube and heated with 5 ml of the acidic ninhydrin reagent for 10 min in a boiling water bath. Sialoglycoproteins are detected as brown bands. No additional procedure such as destaining is necessary. When 20 micrograms fetuin, a sialoglycoprotein, per gel is applied, the band remains visible for at least 2 h. Stained gel can be scanned with a gel scanner at 470 nm. When the stained gel was dried on a sheet of polypropylene filter, the color was stable for at least one month. The present method is superior to the method using Stains-all (3,3'-diethyl-9-methyl-4,5,4',5'-dibenzothiacarbocyanine) in specificity and simplicity for the detection of sialoglycoproteins.</p> No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 44 6 1990 Three-dimensional architecture of rat glomerular basement membrane by ultra-high resolution scanning electron microscopy. 333 335 EN Yasushi Yamasaki Hirofumi Makino Kazue Hironaka Yoshikazu Hayashi Kenichi Shikata Zensuke Ota Article 10.18926/AMO/30431 <p>We demonstrated the ultrastructure of rat glomerular basement membrane (GBM) by ultra-high resolution scanning electron microscopy. GBM prepared by sonication methods and conductive-staining could be observed without metal coating at magnifications as high as 400,000 times. The GBM showed an irregular meshwork structure composed of various strands and pores. The width of the strands ranged from 6 to 15 nm, and the diameter of pores ranged from 6 to 50 nm. The present study confirmed our molecular sieve theory of the basement membrane.</p> No potential conflict of interest relevant to this article was reported.

岡山大学温泉研究所 Acta Medica Okayama 0369-7142 43 1974 北関東山地,単斜輝石かんらん岩の金雲母とパーガス閃石 1 13 EN Koichi Tazaki Michiya Inomata 10.18926/21236 Coexisting interstitial phlogopites and pargasites in iron rich wehrlite from the northern Kanto mountains were studied. The textural relationｓ between those interstitial hydrous minerals and major constituent minerals, such as olivines and clinopyroxenes suggest the crystallization of the former from the intergranular residual liquids. The modal amounts of phlogopites in wehrlite are less than 1%, and that of pargasites range from 2 to 5%. Microprobe analyses have been carried for those hydrous minerals and the major constituent minerals. namely, olivine, clinopyroxene and idiomorphic zoned chromite. Olivines in wehrlite from the northern Kanto mountains have rather iron-rich nature and range from Foso to Fo(80) and those coexisting with the hydrous minerals are Fo(82.5) in average. Clinopyroxenes from mica bearing wehrlite contain about 2.5% Al(2)O(3). 17% MgO and 5.5% FeO. Tentative cation per cent of Ca, Mg and Fe are calculated as Ca(42.6) Mg(48.1) Fe(9.3) in average. The present probe analyses show high magnesian nature of the interstitial phlogopites. In addition, these phlogopites are conspicuous by their high TiO(2) content ranging from 4.6 to 5.2%. Mg/Mg + Fe versus Ti relation plotted in Fig. 4 revealed that the present phlogopite are similar to that of the alkaline pyroxenite nodules from the Lashaine volcano, northern Tanzania. Pargasites in mica bearing wehrlite are also rich in TiO(2) (3.0〜4.7%), whereas pargasitic common hornblende from hornblendite or gabbro are rather low in TiO(2) (0.9〜1.7%). Mg/Mg+Fe+Mn versus Ti relations of amphiboles illustrate that Ti contents increase with decrease of Mg/Mg+Fe +Mn. Two different increasing trend for Ti conten t are also noticed. Idiomorphic chromites show distinct compositional variation from core to rim. Total FeO increases about 16% at rim, whereas TiO(2), Al(2)O(3) and MgO decrease at the rim. Possible crystallization scheme of those hydrous minersls is proposed in Fig. 10, where the maximum stability limits of phlogopite and hornblende are shown with the curve for begining of melting of basalt. Separation of liquid fraction from cumulate phase with a small trapped intergranular liquid might occur at point A. The liquid fraction then might move upward through the course II. In cumulate phase, intergranular liquid might crystallize pargasitic amphibole by the reaction with olivine and clinopyroxene through the course I in Fig. 10. Intergranular liquid might decrease with production of amphiboles and K(2)O might concentrate in residual liquid. At final stage of crystallization, residual liquid rich in K(2)O might crystallize as phlogopite by the reactions with olivine. Decrease of Ti02 content at the nm of idiomorphic chromite might be related to the formations of Ti-rich hydrous minerals. No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0525-1931 33 2 1984 2-ニトロソ-1-ナフトール-4-スルホン酸を用いるイオン対クロマトグラフィーによるコバルトの定量 119 121 EN Ikuo Sawatari Mitsuko Oshima Shoji Motomizu Kyoji Toei Cobalt was spectrophotometrically determined as an anionic metal complex of 2-nitroso-1-naphthol-4-sulfonic acid (nitroso-NW acid) on a reversed-phase high-performance liquid chromatograph. An ODS column(4.6 mm i.d.×100 mm) was used, and the mobile phase consisted of 1.4×10(-2) M tetrabutylammonium bromide and phosphate buffer (pH 8,5×10(-3) M) in a mixture of 52 vol% methanol and 48 vol% distilled water. To 7.5ml of sample solution, 1 ml of citrate buffer solution(2 M, pH 5.4) and 1 ml of nitroso-NW acid(4×10(-3) M) were added. After 10 min, 0.5 ml of 2×10(-3) M EDTA was further added, and the resultant solution was diluted to 10 ml with distilled water. One hundred μl of the solution was injected on to the column. Detection was made on a UV detector at 368 nm. The calibration curve was linear in the range of (125)×10(-7) M of cobalt. By the proposed method, cobalt contents in four commercial nickel salts, Ni(NO(3))(2) · 6H(2)O, NiSO(4) (NH(4))(2) SO(4)·6H(2)O, NiSO(4)·6H(2)O, and NiCl(2)·6H(2)O, were determined to be (0.0030.06)% with relative standard deviation of 1.3% (seven determinations of the nitrate salt). No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0525-1931 33 2 1984 モリブドリン酸によるローダミン6Gの消蛍光を利用するリンの連続流れ定量法 116 119 EN Shoji Motomizu Hiroshi Mikasa Mitsuko Oshima Kyoji Toei Continuous flow method was examined for the determination of phosphorus. The molybdophosphate formed between orthophosphate and molybdate in hydrochloric acid medium diminished the fluorescence of Rhodamine 6 G. Carrier solution (distilled water) and reagent solution were propelled by double plunger pump P(1) and P(2) (flow rate : 0.98 ml/min), and sample solution(160μl) was injected into the carrier stream. The two streams were mixed in 20 cm long Teflon tubing (1 mm i.d.), and the mixture was flowed through a flow cell(18μl), at which the fluoresence of Rhodamine 6 G was detected(λ(ex) =350 nm, λ(em)=580 nm). The reagent solution consists of 0.035 M molybdenum and 1×10(-5) M Rhodamine 6 G in 0.8 M hydrochloric acid. Co-existing ions generally existing in river and sea waters did not interfere the determination of phosphorus. The calibration curve was linear from 0 to 45 ppb of phosphorus. The method was applied to sea water. The sampling rate was 20 samples per hour. No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0525-1931 34 8 1985 フローインジェクション-蛍光法による河川水中のアンモニアの定量 518 521 EN Hiroshi Mikasa Shoji Motomizu Kyoji Toei Flow injection analysis(FIA) was examined for the fluorometric determination of ammonia in river water. Ammonia reacted with ο-phthalaldehyde(OPA) in the presence of 2-mercaptoethanol(ME) to form a fluorescent substance at pH 9.5. The reagent solution containing 10(-2) M OPA and 10(-3) M ME and the carrier fluid(distilled water) were propelled by a double plunger pump at a rate of 1.2 ml/min. The 40μl sample solution, injected into the carrier stream, was mixed with the reagent solution in a Teflon tubing (3 m, 0.5 mm i.d.) and led to a flow cell(18 μl). Fluorescence excited at λ(ex)=350 nm was measured at λ(em)=486 nm. Ions present commonly in river waters did not interfere with the determination of ammonia. An anion exchange column installed just behind a sample injection valve in the flow system was effective in eliminating interferences with amino acids. Using the proposed FIA system, trace amounts of ammonia(3150 ppb as nitrogen) in river water were determined in the rate of 40 samples per h. No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0525-1931 36 3 1987 ニトロソ化反応を利用する硝酸イオンのフローインジェクション/吸光光度法 207 209 EN Cheng Rui Shi Shoji Motomizu Kyoji Toei N, N-Bis (2-hydroxypropyl) aniline (BHPA) reacts with nitrite ion in an acidic medium to form a red product, which has the maximum absorption at 500 nm. On the basis of this color reaction, FIA of nitrate was established. Nitrate ion could be reduced to nitrite ion by passing through the reduction column(2 mm×30 cm) packed with copperized cadmium {Cu(Cd) : particle size, about 0.5 mm}, which was installed just behind the sample injection valve. The carrier solution(10(-3) M EDTA, pH 8) and the reagent solution (8×10(-4) M BHPA, 0.35 M HCl, 0.15 M H(3)PO(4)) were propelled at the flow rate of 0.7 ml/min by using a double-plunger pump. The sample(120 μl) was injected into the carrier stream by using a 6-way injection valve. The reaction coil(0.5 mm×2 m) was kept in a thermostatically controlled water bath(80℃). The cooling coil(0.5 mm×50 cm), which was installed just behind the reaction coil, was kept in a water bath (tap water temperature). A calibration curve was linear up to 2×10(-4) M nitrate. The detection limit corresponding to S/N=2 was 10(-6) M nitrate, and the relative standard deviation (10 injections) was 0.4%, and the sampling rate was 40 samples per hour. Nitrate in river water at concentrations of 10(-5) M level was determined. No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0910-6340 17 11 2001 Highly Sensitive Determination Method for Total Carbonate in Water Samples by Flow Injection Analysis Coupled with Gas-Diffusion Separation 1285 1290 EN Mitsuko Oshima Yanlin Wei Masaki Yamamoto Hiroki Tanaka Toshio Takayanagi Shoji Motomizu A spectrophotometric method for the determination of total carbonate in water samples was developed. The method is based on the color change of an acid-base indicator in relation to the concentration of permeable gas substances through a membrane. By using a new portable FIA system equipped with a gas-diffusion unit, a highly sensitive and on-site determination of total carbonate in aqueous solutions was investigated. A new color-change system with 4-(2',4'-dinitrophenylazo)-1-naphthol-5-sulfonic acid (DNN5S) was developed. Absorbance changes of the reagent solution were measured at 450 nm with a light-emitting diode (LED) as a light source. A new type of gas-diffusion unit was used, and was constructed with double tubing: the inner tubing was a micro porous PTFE (polytetrafluoroethylene) tubing (1.0 mm inner diameter and 1.8 mm outer diameter, pore size 2 μm, porosity 50%); the outer tubing was made of glass with 2.0 mm inner diameter. The optimized system conditions were as follows: the sample size was 200 μl, the temperature of the air bath for the gas-diffusion unit was 25℃, and the length of the gas-diffusion unit was 15 cm; each flow rate was 0.3 ml min(-1). For measuring carbonate at low concentrations, a method for preparing water with less carbonate was proposed: the carbonate content of the water was decreased down to 5×10(-7) M. The calibration graph was rectilinear from 1×10(-6) M to 10(-3) M, and the detection limit (corresponding to a signal-to-noise ratio of 3) was 1×10(-6) M of carbonate. The relative standard deviation (RSD) of ten measurements of 2.3×10(-5) M Na(2)CO(3) solution was 1.9%. The total carbonate in various kinds of water (such as river, sea, rain, distilled and ultra purified) was determined. No potential conflict of interest relevant to this article was reported.

日本分析化学会 Acta Medica Okayama 0525-1931 34 10 1985 2-ニトロソ-5-ジメチルアミノフェノールとゼフィラミンを用いる銅の溶媒抽出-吸光光度定量 643 645 EN Cheng Rui Shi Shoji Motomizu Kyoji Toei 2-Nitroso-5-dimethylaminophenol (nitroso-DMAP) reacts with copper(II) ion to form a 1:2 complex in an aqueous solution. In the presence of zephiramine (tetradecyldimethylbenzylammonium chloride), a 1:3 complex is formed and can be extracted into chloroform. The excess of nitroso-DMAP co-extracted with complex anion into organic phase can be removed by adding an adequate amount of chloride ion at about pH 12. Maximum absorption wavelength of the copper complex in the organic phase is 468 nm, at which the molar absorptivity was found to be 3.6×10(4) l mol(-1) cm(-1). This method was applied to determine micro amounts of copper in iron and steel samples and aluminium alloys. The procedure is as follows. An adequate amount of the sample was dissolved in aqua regia. The resulting solution was evaporated nearly to dryness. The residue was re-dissolved in distilled water to give an adequate volume. Five milliliters of the sample solution were taken into a stoppered test tube. Ascorbic acid and potassium chloride solutions were then added. Dichlorocopper(I) ion was extracted with chloroform solution of zephiramine. The organic phase was shaken with an aqueous solution containing hydrogen peroxide, nitroso-DMAP, potassium chloride, and potassium hydroxide to form the copper complex of nitroso-DMAP and to remove the excess of reagent in the organic phase. The absorbance at 468 nm was measured against the reagent blank. No potential conflict of interest relevant to this article was reported.