Springer Science and Business Media LLC Acta Medica Okayama 0971-5894|0974-0430 2026 Suppression of salt-enhanced apoplastic flow by salicylic acid in rice EN Md. Asadulla Al Galib Graduate School of Environmental and Life Science, Okayama University Maoxiang Zhao Graduate School of Environmental and Life Science, Okayama University Toshiyuki Nakamura Graduate School of Environmental and Life Science, Okayama University Yoshimasa Nakamura Graduate School of Environmental and Life Science, Okayama University Yoshihiko Hirai Graduate School of Environmental and Life Science, Okayama University Yoshitaka Nakashima Graduate School of Environmental and Life Science, Okayama University Shintaro Munemasa Graduate School of Environmental and Life Science, Okayama University Izumi C. Mori Institute of Plant Science and Resources, Okayama University Yoshiyuki Murata Graduate School of Environmental and Life Science, Okayama University Salinity enhances apoplastic flow, resulting in an increment of Na+ uptake and a lower K+/Na+ ratio. Salicylic acid (SA) plays an important role in improving salinity tolerance in plants. The effect of exogenous SA on apoplastic flow in salt-treated rice seedlings was studied using an apoplastic tracer, 8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS) in light. Application of NaCl at 25 mM to the hydroponic solution significantly increased PTS uptake, while 25 mM NaCl did not affect seedling growth. Application of 25 mM NaNO3 increased PTS uptake to the same degree. Salinity significantly increased sodium (Na+) content but had no significant effect on potassium (K+) content, resulting in a lower K+/Na+ ratio. The application of SA at 0.05 mM and 0.1 mM to the hydroponic solution reduced Na-enhanced PTS uptake. Salicylic acid at 0.05 mM and 0.1 mM significantly reduced Na+ content and slightly increased K+ content in the shoots of rice seedlings, resulting in a higher K+/Na+ ratio. However, SA at up to 0.1 mM did not increase SA contents in shoots under salt stress. These results suggest that exogenous SA reduces Na+ uptake by suppressing Na+-enhanced apoplastic flow in rice seedlings. These findings provide insight into modulation of Na+ transport pathways from roots to shoots by SA and may allow us to utilize brackish water for rice cultivation and to improve salt-tolerant rice through suppression of salt-enhanced apoplastic flow by chemicals such as salicylic acid. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1757-4749 18 1 2026 Sodium butyrate augments the antibacterial activity of tetracycline against clinical isolates of multidrug-resistant Vibrio cholerae 9 EN Sushmita Kundu Division of Biochemistry, ICMR- National Institute for Research in Bacterial Infections Sourin Alu Division of Biochemistry, ICMR- National Institute for Research in Bacterial Infections Abhishek Singh Division of Biochemistry, ICMR- National Institute for Research in Bacterial Infections Animesh Gope Division of General Medicine, ICMR- National Institute for Research in Bacterial Infections Ranjan Kumar Nandy Division of Bacteriology, ICMR- National Institute for Research in Bacterial Infections Asish K. Mukhopadhyay Division of Bacteriology, ICMR- National Institute for Research in Bacterial Infections Shin-ichi Miyoshi Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Nabendu Sekhar Chatterjee Division of Biochemistry, ICMR- National Institute for Research in Bacterial Infections Sushmita Bhattacharya Division of Biochemistry, ICMR- National Institute for Research in Bacterial Infections Background Antibiotic resistance poses a major challenge in treating Vibrio cholerae infections. One promising method to counter resistance is the co-administration of antibiotics with non-antibiotic adjuvants to enhance their efficacy. This study investigated the combined action of sodium butyrate (SB) and tetracycline on tetracycline-resistant V. cholerae strains. Results The combined activity of SB and antibiotics was assessed on eight V. cholerae clinical isolates using the Fractional Inhibitory Concentration Index (FICI), with SB-Tetracycline showing strong synergy (FICI: 0.09–0.5). Functional and mechanistic studies, including time-kill kinetics, live/dead staining, SEM-based morphological analysis, and fluorometric assays, demonstrated a synergistic antibacterial effect of SB and Tetracycline. This effect was associated with increased membrane permeability, disruption of membrane integrity, dissipation of the proton motive force, and suppression of efflux activity. These changes collectively led to membrane damage, enhanced intracellular accumulation of Tetracycline, decreased intracellular ATP levels, and ultimately, bacterial cell death. Moreover, GM1-CT ELISA and fluorescence microscopy revealed the synergistic anti-virulence activity of the SB- Tetracycline combination. Finally, the combination of SB and Tetracycline showed enhanced efficacy in animal models compared with monotherapy. Conclusion: The observed SB-Tetracycline synergy provides a promising therapeutic approach to overcome tetracycline resistance in V. cholerae, offering a potential adjunct strategy for the management of antibiotic-resistant cholera infections. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 0041-1132 2026 Pediatric autologous peripheral blood stem cell collection without heparin using a highly concentrated sodium citrate anticoagulant: A retrospective comparison with standard ACD-A EN Keiko Fujii Department of Hematology, Okayama University Hospital Wataru Kitamura Department of Hematology, Okayama University Hospital Kana Washio Department of Pediatrics, Okayama University Hospital Kazuhiro Ikeuchi Department of Hematology, Okayama University Hospital Joji Shimono Department of Hematology, Okayama University Hospital Hiroyuki Murakami Department of Hematology, Okayama University Hospital Fumio Otsuka Division of Clinical Laboratory, Okayama University Hospital Yoshinobu Maeda Department of Hematology, Okayama University Hospital Nobuharu Fujii Department of Hematology, Okayama University Hospital Background: Heparin combined with sodium citrate has been used in leukocytapheresis for pediatric patients. Since 2022, we have performed leukocytapheresis using a highly concentrated sodium citrate solution (HSC, 5.32%) instead of acid citrate dextrose solution A (ACD-A). We conducted this study to determine whether HSC use reduces run time and the total amount of anticoagulant solution in children. Study Design and Methods: We retrospectively analyzed data from consecutive autologous peripheral blood stem cell harvests (auto-PBSCHs) between June 2012 and May 2025, including patient characteristics, mobilization methods, protocol used, anticoagulant type, run time, total anticoagulant solution volume, and collection efficiency. Results: Auto-PBSCH was performed using the mononuclear cell collection (MNC) protocol in 28 procedures and the continuous MNC protocol in 20 procedures. ACD-A was used in 35 procedures and HSC in 13. The run time was significantly shorter (204 [range, 117–302] vs. 157 min [range, 103–227], p = .02) in the HSC group and also confirmed in multivariable regression analysis (coefficient, －55.6; 95% confidence interval, －106.2 to －5.04; p = .03). In a subgroup analysis of cMNC procedures, CD34+ collection efficiency showed a strong negative correlation with the proportion of run time devoted to establishing the initial interface (r = －.73, p = .0003). Conclusion: Delays in establishing the initial interface can reduce the duration of the effective MNC collection phase and may negatively affect collection efficiency. Careful attention to the initial interface phase is therefore warranted when using HSC. No potential conflict of interest relevant to this article was reported.

The Japan Institute of Heterocyclic Chemistry Acta Medica Okayama 0385-5414 69 1 2006 Synthesis of Functionalized Phospholane Oxides and Phosphorinane Oxides from 1,4- and 1,5-Di-O-Mesyloxy Compounds 283 294 EN Tadashi Hanaya Department of Chemistry, Faculty of Science, Okayama University Karsten Schürrle Department of Chemistry, Faculty of Science, Okayama University Hiroshi Yamamoto Department of Chemistry, Faculty of Science, Okayama University Treatment of 1,4-di-O-mesyl-2,3-di-O-methyl-L-threitol (8b) with phenylphosphine in the presence of sodium hydride in DMSO, followed by the action of hydrogen peroxide, afforded 3,4-dimethoxy-1-phenylphospholane 1- oxide (7), while the same treatment of 1,5-di-O-mesyl-2,3,4-tri-O-methyl-meso- xylitol (11b) provided 2,3,4-trimethoxy-1-phenylphosphorinane 1-oxide (14). No potential conflict of interest relevant to this article was reported.

Ceramic Society of Japan Acta Medica Okayama 1348-6535 134 1 2026 Structure and acid resistance of fluoride-treated hydroxyapatite particles 31 37 EN Satoshi Hayakawa Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Kazuki Ando Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Tomohiko Yoshioka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Topical fluoride is used in clinical dentistry to prevent dental caries. The primary reaction product formed on the tooth surface after topical fluoride application is calcium fluoride (CaF2). In this study, we investigated the effect of deposited CaF2 on the acid resistance of fluoride-treated hydroxyapatite (HAp). Commercial HAp particles were treated with two types of fluoride-containing acetic acid-sodium acetate (Ac-AcNa) buffer solutions, with and without the addition of phosphate ions. X-ray diffraction (XRD) and nuclear magnetic resonance (NMR) analyses confirmed the formation of fluorine-substituted hydroxyapatite (F-HAp), along with a hydration layer containing calcium phosphate and CaF2. Phase composition analysis revealed that the presence of phosphate ions in the fluoride treatment solution reduced the amount of CaF2 deposited. Acid resistance evaluation and phase composition analysis indicated that the initial dissolution rate of the fluoride-treated HAp in Ac-AcNa buffer solutions decreased on surfaces densely covered with CaF2 particles. No potential conflict of interest relevant to this article was reported.

Ceramic Society of Japan Acta Medica Okayama 1348-6535 134 1 2026 Preparation of brookite-type titanium dioxide particle layer on titanium surfaces via hydrothermal treatment and evaluation of in vitro apatite-forming ability 24 30 EN Satoshi Hayakawa Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Yushi Nakamoto Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Seiya Kojima Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Noriyuki Nagaoka Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School Takuya Kataoka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University Tomohiko Yoshioka Faculty of Interdisciplinary Science and Engineering in Health Systems, Okayama University In this study, we prepared a brookite-type titanium dioxide particle layer on the surface of titanium substrates via hydrothermal treatment in aqueous urea solutions containing sodium chloride (NaCl) and examined its in vitro apatite-forming ability. Increasing the urea concentration suppressed the formation of anatase-type titanium dioxide on the titanium substrate, forming a particle layer composed of pure brookite-type titanium dioxide. The size and packing density of brookite-type titanium dioxide particles formed on the titanium substrate increased with the NaCl concentration in a 7.0 mol·dm－3 urea solution. When titanium substrates hydrothermally treated in aqueous solutions of 7.0 mol·dm－3 urea and 2.0 mol·dm－3 NaCl were soaked in a simulated body fluid for various periods up to 7 d, the substrate surface was densely covered with hemispherical apatite particles (5.3 µm in diameter) within 3 d, indicating that the brookite-type titanium dioxide particle layer had an excellent apatite-forming ability comparable to that of the anatase-type titanium dioxide particle layer. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0718-9508 2025 Suppression of Na+ Uptake Via Apoplastic Flow by Chitosan in Rice EN Maoxiang Zhao Graduate School of Environmental and Life Science, Okayama University Md. Asadulla Al Galib Graduate School of Environmental and Life Science, Okayama University Toshiyuki Nakamura Graduate School of Environmental and Life Science, Okayama University Yoshimasa Nakamura Graduate School of Environmental and Life Science, Okayama University Yoshihiko Hirai Graduate School of Environmental and Life Science, Okayama University Yoshitaka Nakashima Shintaro Munemasa Graduate School of Environmental and Life Science, Okayama University Izumi C. Mori Institute of Plant Science and Resources, Okayama University Yoshiyuki Murata Graduate School of Environmental and Life Science, Okayama University Purpose: Chitosan enhances tolerance to salinity in rice. Apoplastic flow plays a crucial role in the accumulation of sodium (Na+) in rice under salinity. This study investigated the effects of exogenous chitosan on apoplastic flow and Na+ uptake in NaCl-treated rice seedlings. Methods: We employed an apoplastic tracer, trisodium salt of 8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS), in order to evaluate apoplastic flow in rice (Oryza sativa L., cv. Nipponbare) seedlings that were hydroponically grown in the solution containing NaCl (0 and 25 mM), and chitosan (0 mg L－ 1, 10 mg L－ 1, and 50 mg L－ 1). Results: Application of 25 mM NaCl significantly increased PTS uptake and Na+ content in shoots but did not affect K+ content, resulting in a lower K+/Na+ ratio although 25 mM NaCl did not affect the seedling growth. The application of chitosan suppressed Na+-enhanced PTS uptake and Na+ accumulation in shoots without affecting the K+ content, which led to a higher K+/Na+ ratio. Moreover, chitosan did not affect the reducing sugar content or electrical conductivity in the solution containing NaCl. Conclusions: These results suggest that application of chitosan suppressed Na+-enhanced apoplastic flow to reduce Na+ uptake in rice seedlings. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 6 2025 Effects of Thoron Inhalation and Cyclosporin A Treatment on Dextran Sulfate Sodium-Induced Oxidative Damage in Mice 421 429 EN Ayumi Tanaka Graduate School of Health Sciences, Okayama University Shota Naoe Graduate School of Health Sciences, Okayama University Reiju Takenaka Graduate School of Health Sciences, Okayama University Norie Kanzaki Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency Akihiro Sakoda Ningyo-toge Environmental Engineering Center, Japan Atomic Energy Agency Kiyonori Yamaoka Faculty of Health Sciences, Okayama University Takahiro Kataoka Faculty of Health Sciences, Okayama University Original Article 10.18926/AMO/69844 Radon (222Rn; Rn) and thoron (220Rn; Tn) inhalation have been reported to enhance antioxidant activity in various organs. However, the effects of Tn on the colon have not been investigated. This study aimed to clarify the effects of Tn inhalation, alone and in combination with cyclosporin A (CsA), on dextran sulfate sodium (DSS)-induced colitis, and the accompanying oxidative stress, in mice. Male BALB/c mice were subjected to continuous 8-day Tn inhalation (c-Tn, 533±128 Bq/m3) or alternate-day Tn inhalation over the same period (f-Tn, 577±63Bq/m3), followed by treatment with 3% DSS and either CsA or vehicle for 7 days. Although the disease activity index (DAI) decreased significantly by day 2 in the c-Tn group, scores remained significantly higher than those in the f-Tn group. In the c-Tn group, superoxide dismutase and catalase activity in the colon were significantly elevated compared with those in sham controls. Thus, DSS-induced damage was effectively inhibited in the earlier stages by the c-Tn mode of inhalation than by the f-Tn mode. These findings suggest that continuous Tn inhalation more effectively attenuated early colitis symptoms than alternate-day inhalation, potentially through upregulation of antioxidant defenses. Tn and CsA showed no combined effects. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 0960-7412 120 1 2024 OsHAK4 functions in retrieving sodium from the phloem at the reproductive stage of rice 76 90 EN Jing Che Institute of Plant Science and Resources, Okayama University Naoki Yamaji Institute of Plant Science and Resources, Okayama University Shao Fei Wang Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences Yue Xia Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences Shun Ying Yang Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences Yan Hua Su Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences Ren Fang Shen Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences Jian Feng Ma Institute of Plant Science and Resources, Okayama University Soil salinity significantly limits rice productivity, but it is poorly understood how excess sodium (Na+) is delivered to the grains at the reproductive stage. Here, we functionally characterized OsHAK4, a member of the clade IV HAK/KUP/KT transporter subfamily in rice. OsHAK4 was localized to the plasma membrane and exhibited influx transport activity for Na+, but not for K+. Analysis of organ- and growth stage-dependent expression patterns showed that very low expression levels of OsHAK4 were detected at the vegetative growth stage, but its high expression in uppermost node I, peduncle, and rachis was found at the reproductive stage. Immunostaining indicated OsHAK4 localization in the phloem region of node I, peduncle, and rachis. Knockout of OsHAK4 did not affect the growth and Na+ accumulation at the vegetative stage. However, at the reproductive stage, the hak4 mutants accumulated higher Na+ in the peduncle, rachis, husk, and brown rice compared to the wild-type rice. Element imaging revealed higher Na+ accumulation at the phloem region of the peduncle in the mutants. These results indicate that OsHAK4 plays a crucial role in retrieving Na+ from the phloem in the upper nodes, peduncle, and rachis, thereby preventing Na+ distribution to the grains at the reproductive stage of rice. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 1473-0502 64 6 2025 Novel leukocytapheresis method using highly concentrated sodium citrate solution for the manufacturing of tisagenlecleucel 104265 EN Wataru Kitamura Department of Hematology and Oncology, Okayama University Hospital Keiko Fujii Department of Hematology and Oncology, Okayama University Hospital Masaya Abe Department of Hematology and Oncology, Okayama University Hospital Kazuhiro Ikeuchi Department of Hematology and Oncology, Okayama University Hospital Kana Washio Department of Pediatrics, Okayama University Hospital Fumio Otsuka Division of Clinical Laboratory, Okayama University Hospital Yoshinobu Maeda Department of Hematology and Oncology, Okayama University Hospital Nobuharu Fujii Department of Hematology and Oncology, Okayama University Hospital For the manufacturing of tisagenlecleucel (tisa-cel) requires the non-mobilized mononuclear cell collection (MNC). CD3+ cell collection is performed using the same protocol as autologous peripheral blood stem cell harvest (auto-PBSCH), but this procedure necessitates the same target CD3+ cell yields regardless of age or body weight, which may take several days especially in pediatric and small female patients with low white blood cell counts. We previously demonstrated a novel method using highly concentration sodium citrate (HSC), which reduced the need for an anticoagulant (AC) solution and shortened the procedure time in auto-PBSCH. This novel method was expected to offer advantages for smaller patients, prompting us to investigate its application in leukocytapheresis for the manufacturing of tisa-cel. We retrospectively analyzed consecutive leukocytapheresis data obtained using Spectra Optia continuous MNC mode between November 2022 and June 2024 at our institution (n = 9). In six of nine patients, pre-leukocytapheresis CD3+ cell counts were less than 500 /μL, but all could obtain the target CD3+ cell yields in one day upon processing blood volume adjustment. When we compared patients who had received CD3+ cell collection using normal-concentration sodium citrate (NSC) as our previously reported using propensity score-matched pair analysis, the total AC solution volume was significantly lower (1168 vs. 316 mL, p < 0.001) and procedure time was significantly shorter (254 vs. 228 min, p = 0.04) in the HSC group compared to the NSC group. In conclusion, this procedure was also useful for non-mobilized MNC. Our findings warrant validation in a larger patient cohort. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 0041-1132 65 9 2025 Novel method for autologous peripheral blood stem cell harvest using highly concentrated sodium citrate solution replacing acid citrate dextrose solution A 1662 1672 EN Wataru Kitamura Department of Hematology and Oncology, Okayama University Hospital Keiko Fujii Department of Hematology and Oncology, Okayama University Hospital Masaya Abe Department of Hematology and Oncology, Okayama University Hospital Kazuhiro Ikeuchi Department of Hematology and Oncology, Okayama University Hospital Joji Shimono Department of Hematology and Oncology, Okayama University Hospital Kana Washio Department of Pediatrics, Okayama University Hospital Fumio Otsuka Division of Clinical Laboratory, Okayama University Hospital Yoshinobu Maeda Department of Hematology and Oncology, Okayama University Hospital Nobuharu Fujii Department of Hematology and Oncology, Okayama University Hospital Background: As the processed blood volume increases, a larger amount of anticoagulant (AC) is required, which leads to a serious issue of fluid dilution in large-volume leukocytapheresis (defined as ≥3-fold total blood volume). We previously reported a novel method for allogeneic peripheral blood stem cell harvest (PBSCH) using highly concentrated sodium citrate (HSC; 5.32%), which shortened the procedure time and reduced the need for an AC solution without heparin. In this study, we extended this novel method to autologous PBSCH (auto-PBSCH) and compared it with patients who received auto-PBSCH using normal concentrated sodium citrate (NSC; 2.2%). Study Design and Methods: We retrospectively analyzed consecutive auto-PBSCH data obtained using the Spectra Optia continuous mononuclear cell collection mode between May 2017 and May 2025 at our institution. Results: Leukocytapheresis was performed using NSC in 36 patients and HSC in 22. In the HSC group, patients tended to be younger, had significantly lower body weight, and had significantly fewer hematopoietic tumors as primary diseases compared to the NSC group. After propensity score-matched cohort adjusted for patient background, the total amount of AC solution was significantly lower (694 [range, 77–1648] vs. 298 mL [range, 64–797], p = .02), and procedure time was significantly shorter (224 [range, 117–395] vs. 181 min [range, 103–309], p = .048) in the HSC group. Furthermore, the loss rates of magnesium and potassium were lower in the HSC group. Conclusion: This novel leukocytapheresis method demonstrated the efficacy and safety in auto-PBSCH, while minimizing the patient burden. No potential conflict of interest relevant to this article was reported.

Tech Science Press Acta Medica Okayama 1851-5657 94 11 2025 Survey of Barley Sodium Transporter HvHKT1;1 Variants and Their Functional Analysis 3653 3665 EN Shahin Imran Institute of Plant Science and Resources, Okayama University Maki Katsuhara Department of Agronomy, Khulna Agricultural University Barley (Hordeum vulgare L.) employs the Na+ transporter HvHKT1;1, which is an N+-selective transporter. This study characterized the full-length HvHKT1;1 (HvHKT1;1-FL) and three mRNA variants (HvHKT1;1-V1, -V2, and -V3), which encode polypeptides of 64.7, 54.0, 40.5, and 32.9 kDa, respectively. Tissue-specific expression profiling revealed that HvHKT1;1-FL is the most abundant transcript across leaf, sheath, and root tissues under normal conditions, with the highest expression in leaves. Under 150 mM NaCl stress, HvHKT1;1-FL and its variants showed a dynamic, time-dependent expression pattern, with peak leaf expression at 2 h, sheath expression at 12 h, and root expression at 2 h, suggesting their roles in early stress response. Functional analysis using two-electrode voltage-clamp measurements demonstrated that HvHKT1;1-FL is highly selective for Na+, with minimal conductance for K+, Li+, Rb+, or Cs+. It demonstrated high Na+ transport efficiency, characterized by higher Vmax and lower Km values, while the variants showed reduced Na+ currents, lower Vmax, and higher Km values, indicating decreased Na+ transport capacity. Reversal potential analyses further confirmed Na+ selectivity, with HvHKT1;1-FL displaying the strongest preference for Na+. Notably, while all variants retained Na+ selectivity, they showed reduced efficiency, as indicated by a more negative reversal potential in low Na+ conditions. These findings highlight the functional diversity among HvHKT1;1 variants, with HvHKT1;1-FL playing a dominant role in Na+ transport. The tissue-specific regulation of these variants under salinity stress underscores their importance in barley’s adaptive responses. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0916-9636 48 11 2025 Efficacy and safety of esaxerenone with and without sodium–glucose cotransporter-2 inhibitor use in hypertensive patients with type 2 diabetes mellitus: a pooled analysis of five clinical studies 2924 2937 EN Hirohiko Motoki Department of Cardiovascular Medicine, Shinshu University School of Medicine Koichiro Kuwahara Department of Cardiovascular Medicine, Shinshu University School of Medicine Haruhito A. Uchida Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Jun Wada Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Kazuomi Kario Division of Cardiovascular Medicine, Department of Medicine, Jichi Medical University School of Medicine Tomohiro Katsuya Katsuya Clinic Tatsuo Shimosawa Department of Clinical Laboratory, School of Medicine, International University of Health and Welfare Kenichi Tsujita Department of Cardiovascular Medicine, Graduate School of Medical Sciences, Kumamoto University Shoko Suzuki Data Intelligence Department, Daiichi Sankyo Co. Ltd. Tomohiro Suedomi Primary Medical Science Department, Daiichi Sankyo Co. Ltd. Takashi Taguchi Primary Medical Science Department, Daiichi Sankyo Co. Ltd. This pooled subanalysis of five multicenter, prospective, open-label, single-arm studies on esaxerenone aimed to evaluate the efficacy, organ-protective effects, and safety of esaxerenone in hypertensive patients with type 2 diabetes mellitus (T2DM), with and without concomitant sodium–glucose cotransporter-2 inhibitor (SGLT2i) therapy. In total, 283 and 279 patients were included in the safety (with SGLT2i, 148; without, 135) and full analysis sets (with SGLT2i; 145; without, 134), respectively. Significant changes in morning home systolic/diastolic blood pressure (SBP/DBP) from baseline to Week 12 were shown in the overall population (mean change: －11.9/－5.2 mmHg, both P < 0.001) and both SGLT2i and non-SGLT2i subgroups (－11.3/－4.8 and －12.5/－5.7 mmHg, respectively, all P < 0.001). Similar findings were observed in bedtime home and office SBP/DBP. The proportions of patients who achieved target home SBP/DBP < 135/85 mmHg were 71.2% (overall population) and 70.5% and 71.9% in the SGLT2i and non-SGLT2i subgroups, respectively. The urine albumin-to-creatinine ratio significantly improved from baseline to Week 12 in the overall population and SGLT2i subgroups (percentage change in geometric mean from baseline: －42.8%, －43.0%, and －42.6%, respectively, all P < 0.001). N-terminal pro-B-type natriuretic peptide levels improved in all groups. The incidence of serum potassium ≥5.5 mEq/L was 2.0% vs 5.2% in the SGLT2i vs non-SGLT2i subgroups. Esaxerenone demonstrated significant BP-lowering effects, and improved renal and cardiovascular parameters, regardless of SGLT2i use. Safety was consistent across groups, with the numerically lower incidence of serum potassium ≥5.5 mEq/L in the SGLT2i subgroup suggesting a potential mitigating effect of SGLT2is on the risk of hyperkalemia. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 0916-9636 48 9 2025 Efficacy and safety of esaxerenone in hypertensive patients with chronic kidney disease, with or without type 2 diabetes mellitus: a pooled analysis of five clinical studies 2413 2426 EN Haruhito A. Uchida Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Jun Wada Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences Hirohiko Motoki Department of Cardiovascular Medicine, Shinshu University School of Medicine Koichiro Kuwahara Department of Cardiovascular Medicine, Shinshu University School of Medicine Kazuomi Kario Division of Cardiovascular Medicine, Department of Medicine, Jichi Medical University School of Medicine Tomohiro Katsuya Katsuya Clinic Tatsuo Shimosawa Department of Clinical Laboratory, School of Medicine, International University of Health and Welfare Kenichi Tsujita Department of Cardiovascular Medicine, Graduate School of Medical Sciences, Kumamoto University Shoko Suzuki Data Intelligence Department, Daiichi Sankyo Co., Ltd. Tomohiro Suedomi Primary Medical Science Department, Daiichi Sankyo Co., Ltd. Takashi Taguchi Primary Medical Science Department, Daiichi Sankyo Co., Ltd. Effective management of blood pressure (BP) and albuminuria are crucial for suppressing chronic kidney disease (CKD) progression and cardiovascular risks in hypertension. This pooled analysis evaluated the antihypertensive effects, organ-protective effects, and safety of esaxerenone in hypertensive patients with CKD by integrating five clinical studies of esaxerenone. Patients were divided based on type 2 diabetes mellitus (T2DM) status (with or without T2DM) and creatinine-based estimated glomerular filtration rate (eGFRcreat) (30 to <60 and ≥60 mL/min/1.73 m2). Significant changes in morning home BP from baseline at Week 12 were observed in the overall population (mean change －12.8/ － 5.4 mmHg), T2DM subgroups ( － 12.2/ － 4.5 and －14.5/ － 7.8 mmHg), and eGFRcreat subgroups ( － 12.5/ － 4.7 and －14.0/ － 6.9 mmHg) (all P < 0.001). Bedtime home and office BP showed similar tendencies. Urine albumin-to-creatinine ratio significantly improved from baseline at Week 12 in the overall population (mean change: －55.2%), T2DM subgroups ( － 56.5% and －52.0%), and eGFRcreat subgroups ( － 54.6% and －55.4%) (all P < 0.001). N-terminal pro-B-type natriuretic peptide levels significantly decreased in the overall population (percent change: －14.1%) and subgroup without T2DM ( － 25.3%). The incidence of serum potassium ≥5.5 mEq/L was lower in the subgroup with T2DM vs without T2DM (3.1% and 11.3%), potentially related to the use of sodium–glucose cotransporter 2 inhibitors. These findings highlight the sustained BP-lowering effect of esaxerenone throughout the day in hypertensive patients with CKD, irrespective of T2DM status, and its significant reduction in albuminuria. The data support the safety and efficacy of esaxerenone in this patient population, underscoring its potential as a valuable therapeutic option. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 2168-8184 17 10 2025 Bladder Trigone as a Sensory Hub: A Narrative Review e94951 EN Takuya Sadahira 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 Takanori Sekito Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomofumi Watanabe 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 The bladder trigone is an anatomically and functionally distinct region within the lower urinary tract (LUT), characterized by a dense network of afferent sensory fibers, specialized urothelial interactions, and prominent mechanotransduction mechanisms. Its intricate neuroarchitecture enables precise detection of bladder filling and coordination of micturition, whereas dysregulation of these pathways contributes to lower urinary tract symptoms (LUTS), including urgency, frequency, and bladder pain. Despite its recognized clinical relevance, the structural and functional basis of trigonal sensory signaling - and its role - remain incompletely understood. This review synthesizes current evidence on trigonal afferent organization, integrating data from anatomical mapping, receptor profiling, electrophysiological characterization, and translational research. Seminal anatomical observations are combined with recent advances in mechanotransduction and purinergic, peptidergic, and transient receptor potential (TRP) signaling to provide a comprehensive perspective. The trigone exhibits three principal afferent classes: (1) intraepithelial fibers penetrating umbrella cells, marked by P2X purinoceptor 3 (P2X3), transient receptor potential vanilloid 1 (TRPV1), calcitonin gene-related peptide (CGRP), and substance P (SP); (2) subepithelial plexuses surrounding microvasculature, enriched in vasoactive neuropeptides and exhibiting plastic hypertrophy in overactive bladder (OAB) and interstitial cystitis/bladder pain syndrome (IC/BPS); and (3) encapsulated corpuscular endings at the lamina propria-detrusor junction, expressing PIEZO1/2 and acid-sensing ion channels (ASICs) for rapid adaptation. In trigeminal dorsal root ganglion (DRG) neurons, high expression of PIEZO2, P2RX3, and voltage-gated sodium channel, type 1.8 (Nav1.8) was observed, revealing their role as the foundation for multisensory information processing. Functional assays highlight distinct mechanotransductive and chemosensory pathways, with aging, inflammation, and neurotrophic factors driving afferent plasticity underlying abnormal bladder sensation, such as urgency, frequency, and pain. Early clinical trials of P2X3 antagonists and intravesical TRPV1 inhibitors demonstrate promising symptomatic benefits. Collectively, evidence positions the bladder trigone as a critical sensory hub where neuronal, urothelial, and immune signals converge to regulate bladder sensation. Understanding its molecular and structural specialization may inform the development of region-specific neuromodulatory therapies targeting sensory urgency and afferent-driven bladder dysfunction. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 79 5 2025 Blood Pressure and Heart Rate Patterns Identified by Unsupervised Machine Learning and Their Associations with Subclinical Cerebral and Renal Damage in a Japanese Community: The Masuda Study 369 379 EN Takashi Hisamatsu Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Minako Kinuta Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Sosuke Munetomo Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mari Fukuda Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Katsuhide Kojima Department of Radiology, Okayama University Hospital Kaori Taniguchi Department of Environmental Medicine and Public Health, Izumo, Shimane University Faculty of Medicine Noriko Nakahata Department of Health and Nutrition, The University of Shimane Faculty of Nursing and Nutrition Hideyuki Kanda Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/69438 We applied unsupervised machine learning to analyze blood pressure (BP) and resting heart rate (HR) patterns measured during a 1-year period to assess their cross-sectional relationships with subclinical cerebral and renal target damage. Dimension reduction via uniform manifold approximation and projection, followed by K-means++ clustering, was used to categorize 362 community-dwelling participants (mean age, 56.2 years; 54.9% women) into three groups: Low BP and Low HR (Lo-BP/Lo-HR), High BP and High HR (Hi-BP/Hi-HR), and Low BP and High HR (Lo-BP/Hi-HR). Cerebral vessel lesions were defined as the presence of at least one of the following magnetic resonance imaging findings: lacunar infarcts, white matter hyperintensities, cerebral microbleeds, or intracranial artery stenosis. A high urinary albumin-to-creatinine ratio (UACR) was defined as the top 10% (≥ 12 mg/g) of the mean value from ≥2 measurements. Poisson regression with robust error variance, adjusted for demographics, lifestyle, and medical history, showed that the Hi-BP/Hi-HR group had relative risks of 3.62 (95% confidence interval, 1.75-7.46) for cerebral vessel lesions and 3.58 (1.33-9.67) for high UACR, and the Lo-BP/Hi-HR group had a relative risk of 3.09 (1.12-8.57) for high UACR, compared with the Lo-BP/Lo-HR group. These findings demonstrate the utility of an unsupervised, data-driven approach for identifying physiological patterns associated with subclinical target organ damage. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 1572-1000 54 2025 Near-infrared photoimmunotherapy for recurrent cancer at the base of the tongue 104719 EN Takuma Makino Department of Otolaryngology - Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Asami Nishikori Department of Hematopathology, Okayama University Graduate School of Health Sciences Yasuharu Sato 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 Junya Matsumoto Department of Otolaryngology - Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shohei Fujimoto 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 Near-infrared photoimmunotherapy (NIR-PIT) is a novel therapeutic approach that targets epidermal growth factor receptor (EGFR). In NIR-PIT, administration of cetuximab sarotalocan sodium is followed by laser irradiation of the affected area, which theoretically should induce tumor cell death. However, residual tumors are occasionally observed. This study investigated factors that influence the therapeutic efficacy of NIR-PIT in cases of recurrence of cancer at the base of the tongue. Six patients undergoing 11 treatment cycles were analyzed, focusing on the puncture interval of cylindrical diffusers and the expression of EGFR in tumors. The results demonstrated that a puncture interval of ≤12 mm significantly enhanced therapeutic efficacy, with one case achieving complete response. EGFR expression was positive in all cases and expression score showed no significant change between before and after treatment. These findings suggest that puncture interval plays a critical role in therapeutic outcomes, whereas EGFR expression may not directly influence treatment efficacy. No potential conflict of interest relevant to this article was reported.

Elsevier Acta Medica Okayama 0032-5791 104 3 2025 An ultra-simplified protocol for PCR template preparation from both unsporulated and sporulated Eimeria oocysts 104810 EN Aruto Takano Departments of Veterinary Immunology, Graduate School of Veterinary Medical Sciences, Osaka Metropolitan University Dennis V. Umali Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, College April H. Wardhana Research Center for Veterinary Science, National Research and Innovation Agency Dyah H. Sawitri Research Center for Veterinary Science, National Research and Innovation Agency Isao Teramoto Departments of Virology and Parasitology, Graduate School of Medicine, Osaka Metropolitan University Toshimitsu Hatabu Laboratory of Animal Physiology, Graduate School of Environmental, Life, Natural Science and Technology, Okayama University Yasutoshi Kido Departments of Virology and Parasitology, Graduate School of Medicine, Osaka Metropolitan University Akira Kaneko Departments of Virology and Parasitology, Graduate School of Medicine, Osaka Metropolitan University Kazumi Sasai Departments of Veterinary Immunology, Graduate School of Veterinary Medical Sciences, Osaka Metropolitan University Hiromitsu Katoh Departments of Veterinary Immunology, Graduate School of Veterinary Medical Sciences, Osaka Metropolitan University Makoto Matsubayashi Departments of Veterinary Immunology, Graduate School of Veterinary Medical Sciences, Osaka Metropolitan University Molecular biological techniques have enabled the accurate identification of the avian Eimeria parasite, however, the preparation of PCR template remains a bottleneck due to contaminants from feces and the robust oocyst's wall resistant to chemical and mechanical force. Generally, the preparation of PCR template involves three main steps: (1) pretreatment of oocysts; (2) disruption of oocysts; and (3) purification of genomic DNA. We prepared PCR templates from both unsporulated and sporulated E. tenella oocysts using various protocols, followed by species-specific PCR to define the limit of detection. Our data revealed that whereas neither pretreatment of oocysts with sodium hypochlorite nor purification of genomic DNA with commercial kits improved the limit of detection of PCR, disruption of oocysts was a critical step in the preparation of PCR templates. The most sensitive PCR assay was achieved with the template prepared by disrupting oocysts suspended in distilled water, followed by bead-beating and heating at 99°C for 5 min, which detected 0.16 oocysts per PCR. This ultra-simplified protocol for preparation of PCR template, which does not require expensive reagents or equipment, will significantly enhance the sensitive and efficient molecular identification of Eimeria. It will improve our understanding of the prevalence of this parasite at the species level and contribute to the development of techniques for the control in the field. 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.

Wiley Acta Medica Okayama 0041-1132 2025 Novel method of leukocytapheresis using a highly concentrated sodium citrate solution alternative to acid citrate dextrose solution A EN Masaya Abe Division of Transfusion and Cell Therapy, Okayama University Hospital Keiko Fujii Division of Clinical Laboratory, Okayama University Hospital Toshiharu Mitsuhashi Center for Innovative Clinical Medicine, Okayama University Hospital Wataru Kitamura Division of Transfusion and Cell Therapy, Okayama University Hospital Kazuhiro Ikeuchi Division of Transfusion and Cell Therapy, Okayama University Hospital Takuya Fukumi Division of Transfusion and Cell Therapy, Okayama University Hospital Kana Washio Department of Pediatric Hematology and Oncology, Okayama University Hospital Fumio Otsuka Division of Clinical Laboratory, Okayama University Hospital Yoshinobu Maeda Department of Hematology and Oncology, Okayama University Hospital Nobuharu Fujii Division of Transfusion and Cell Therapy, Okayama University Hospital Background: Large-volume leukocytapheresis is time consuming. The upper limit of the inlet flow rate is determined by the inlet: anticoagulant (AC) ratio and can be changed by combining the AC with heparin. Here, we devised a protocol to increase the AC ratio using a highly concentrated sodium citrate solution without heparin. Study Design and Methods: We collected data from 40 consecutive apheresis procedures performed using the Spectra Optia system on 40 donors for allogeneic peripheral blood stem cells between June 2022 and June 2023. We used AC containing 2.2% sodium citrate (normal concentrated sodium citrate [NSC]) and 5.32% sodium citrate (highly concentrated sodium citrate [HSC]). The AC ratios were set to 12:1 and 24:1 for the NSC and HSC, respectively. Results: The processed volume was not different; the maximum inlet flow rate increased, the total processing time was reduced, the AC solution used was reduced, and the product volume was reduced in the HSC group, compared to the NSC group. Although the CD34+ cell CE2 was reduced in the HSC group, no difference was observed in the number of collected CD34+ cells. The incidences of citrate-related reactions were similar. Discussion: We propose a novel leukocytapheresis method using HSC that shortens the procedure time and reduces the amount of AC solution used compared to the conventional method No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2227-9059 13 2 2025 The Abnormal Expression of Tubular SGLT2 and GULT2 in Diabetes Model Mice with Malocclusion-Induced Hyperglycemia 267 EN Koichiro Kajiwara Department of Oral Growth & Development, Fukuoka Dental College Sachio Tamaoki Department of Oral Growth & Development, Fukuoka Dental College Yoshihiko Sawa Department of Oral Function & Anatomy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Background: A relationship between malocclusion and the promotion of diabetes has been suggested. In hyperglycemia, the expression of sodium-glucose cotransporter 2 (SGLT2) and the facilitative glucose transporter 2 (GLUT2) is upregulated in proximal tubular cells, leading to an increase in renal glucose reabsorption. The present study aimed to investigate whether malocclusion contributes to diabetic exacerbation. Methods: Streptozotocin (STZ)-induced diabetic mice with malocclusion due to cutting molars were investigated based on increased blood glucose levels. PCR and immunohistochemical analyses were performed on diabetic mice kidneys to investigate the expression of SGLT2 and GLUT2. Results: Animal experiments were performed using 32 mice for 21 days. The time to reach a diabetic condition in STZ-administered mice was shorter with malocclusion than without malocclusion. The increase and mean blood glucose levels in STZ-administered mice were steeper and higher with malocclusion than without malocclusion. Urea albumin, BUN, and CRE levels were higher in diabetic mice with malocclusion than in diabetic mice without. Immunoreaction with anti-SGLT2 and anti-GLUT2 in the renal tissue of STZ-administered mice was stronger with malocclusion than without malocclusion. The amounts of SGLT2 and GLUT2 mRNA in the renal tissue in STZ-administered mice were higher with malocclusion than without malocclusion. The amounts of TNF-a and IL-6 mRNA in the large intestinal tissue in STZ-administered mice were higher with malocclusion than without malocclusion. Conclusions: Our results indicate that malocclusion accelerates the tubular expression of SGLT2 and GLUT2 under hyperglycemia. Malocclusion may be a diabetes-exacerbating factor with increased poor glycemic control due to shortened occlusion time resulting from swallowing food without chewing. No potential conflict of interest relevant to this article was reported.

BMC Acta Medica Okayama 1471-2369 26 1 2025 Exacerbation of diabetes due to F. Nucleatum LPS-induced SGLT2 overexpression in the renal proximal tubular epithelial cells 38 EN Aiko Seki Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Koichiro Kajiwara Department of Oral Growth & Development, Fukuoka Dental College Jumpei Teramachi Department of Oral Function & Anatomy, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Masahiko Egusa Department of Dental Anesthesiology & Special Care Dentistry, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Takuya Miyawaki Department of Dental Anesthesiology & Special Care Dentistry, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Yoshihiko Sawa Department of Oral Function & Anatomy, Dentistry and Pharmaceutical Sciences, Okayama University Graduate School of Medicine Background Diabetes treatments by the control of sodium-glucose cotransporter 2 (SGLT2) is commonly conducted while there are still uncertainties about the mechanisms for the SGLT2 overexpression in kidneys with diabetes. Previously, we have reported that glomeruli and proximal tubules with diabetic nephropathy express toll-like receptor TLR2/4, and that the TLR ligand lipopolysaccharide (LPS) of periodontal pathogens have caused nephropathy in diabetic model mice. Recently, many researchers suggested that the periodontal pathogenic bacteria Fusobacterium (F.) nucleatum has the TLR4-associated strong activator of the colorectal inflammation and cancer. The present study aimed to investigate the possibility of F. nucleatum as an exacerbation factor of diabetes through the renal SGLT2 induction. Methods The induction of the SGLT2 by F. nucleatum LPS (Fn-LPS) were investigated in the streptozotocin-induced diabetic mouse renal tissue and cultured renal proximal epithelial cells. The changes of blood glucose levels and survival curves in diabetic mice with Fn-LPS were analyzed. The Fn-LPS-induced SGLT2 production in the diabetic mouse renal tissue and in the cultured proximal epithelial cells was examined by ELISA, quantitative RT-PCR, and immunohistochemical analysis. Results The SGLT2 expression in the cultured mouse tubular epithelial cells was significantly increased by TNF- or co-culture with Fn-LPS-supplemented J774.1 cells. The period to reach diabetic condition was significantly shorter in Fn-LPS-administered diabetic mice than in diabetic mice. All Fn-LPS-administered-diabetic mice reached humane endpoints during the healthy period of all of the mice administered Fn-LPS only. The promotion of the SGLT2 expression at the inner lumen of proximal tubules were stronger in the Fn-LPS-administered-diabetic mice than in diabetic mice. The renal tissue SGLT2 mRNA amounts and the number of renal proximal tubules with overexpressed SGLT2 in the lumen were more in the Fn-LPS-administered-diabetic mice than in diabetic mice. Conclusions This study suggests that F. nucleatum causes the promotion of diabetes through the overexpression of SGLT2 in proximal tubules under the diabetic condition. Periodontitis with F. nucleatum may be a diabetic exacerbating factor. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2024 胆汁酸塩をco-formerとしたbrick dust AntiY5R の共非晶質化と溶解性改善 EN Shohei AIKAWA Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2024 クロスカルメロースナトリウムの塩基性環境下における高吸水性ゲル化を利用した新規徐放化システムの開発 EN Masato GOMI Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0085-2538 106 5 2024 A randomized, open-label, clinical trial examined the effects of canagliflozin on albuminuria and eGFR decline using an individual pre-intervention eGFR slope 972 984 EN Satoshi Miyamoto Center for Innovative Clinical Medicine, Okayama University Hospital Hiddo J.L. Heerspink Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen Dick de Zeeuw Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen Kota Sakamoto Center for Innovative Clinical Medicine, Okayama University Hospital Michihiro Yoshida Center for Innovative Clinical Medicine, Okayama University Hospital Masao Toyoda Division of Nephrology, Endocrinology and Metabolism, Department of Internal Medicine, Tokai University School of Medicine Daisuke Suzuki Suzuki Diadetes Clinic Takashi Hatanaka Department of Diabetes and Endocrinology, National Hospital Organization Fukuyama Medical Center Tohru Nakamura Diabetes Internal Medicine, Sumitomo Besshi Hospital Shinji Kamei Department of Diabetic Medicine, Kurashiki Central Hospital Satoshi Murao Department of Diabetes and Endocrinology, Takamatsu Hospital Kazuyuki Hida Department of Diabetology and Metabolism, National Hospital Organization Okayama Medical Center Shinichiro Ando Department of Internal Medicine Diabetic Center, Okayama City Hospital Hiroaki Akai Division of Diabetes and Metabolism, Faculty of Medicine, Tohoku Medical and Pharmaceutical University Yasushi Takahashi Department of Diabetes, Ochiai General Hospital Munehiro Kitada Department of Diabetology and Endocrinology, Kanazawa Medical University Hisashi Sugano Department of Diabetes and Endocrinology, Kochi Health Sciences Center Tomokazu Nunoue Nunoue Clinic Akihiko Nakamura Internal Medicine, Osafune Clinic Motofumi Sasaki Department of Diabetes and Endocrinology, Matsue City Hospital Tatsuaki Nakatou Diabetes Center, Okayama Saiseikai General Hospital Kei Fujimoto Division of Diabetes, Metabolism and Endocrinology, Department of Internal Medicine, The Jikei University Kashiwa Hospital Daiji Kawanami Department of Endocrinology and Diabetes, Fukuoka University School of Medicine Takashi Wada Department of Nephrology and Laboratory Medicine, Graduate School of Medical Sciences, Kanazawa University Nobuyuki Miyatake Department of Hygiene, Faculty of Medicine, Kagawa University Hiromi Kuramoto Center for Innovative Clinical Medicine, Okayama University Hospital Kenichi Shikata Center for Innovative Clinical Medicine, Okayama University Hospital Demonstrating drug efficacy in slowing kidney disease progression requires large clinical trials when targeting participants with an early stage of chronic kidney disease (CKD). In this randomized, parallel-group, open-labeled trial (CANPIONE study), we assessed the effect of the sodium-glucose cotransporter 2 (SGLT2) inhibitor canagliflozin using the individual’s change in estimated glomerular filtration rate (eGFR) slope before (pre-intervention slope) and during treatment (chronic slope). We randomly assigned (1:1) participants with type 2 diabetes, urinary albumin-to-creatinine ratio (UACR) of 50 to under 300 mg/g, and an eGFR of at least 45 ml/min/1.73m2 to receive canagliflozin or guideline-recommended treatment except for SGLT2 inhibitors (control). The first and second primary outcomes were the geometric mean percentage change from baseline in UACR and the change in eGFR slope, respectively. Of 98 randomized participants, 96 received at least one study treatment. The least-squares mean change from baseline in log-transformed geometric mean UACR was significantly greater in the canagliflozin group than the control group (between group-difference, －30.8% (95% confidence interval －42.6 to －16.8). The between-group difference (canagliflozin group – control group) of change in eGFR slope (chronic – pre-intervention) was 4.4 (1.6 to 7.3) ml/min/1.73 m2 per year, which was more pronounced in participants with faster eGFR decline. In summary, canagliflozin reduced albuminuria and the participant-specific natural course of eGFR decline in participants with type 2 diabetes and microalbuminuria. Thus, the CANPIONE study suggests that the within-individual change in eGFR slope may be a novel approach to determine the kidney protective potential of new therapies in early stages of CKD. No potential conflict of interest relevant to this article was reported.

Nature Portfolio Acta Medica Okayama 2045-2322 14 1 2024 Effects of dapagliflozin on myoglobin efflux from cardiomyocyte during myocardial ischemia/reperfusion in anesthetized rats 16337 EN Tomohiro Hayashida Department of Cardiovascular Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yosuke Kuroko Department of Cardiovascular Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences and Okayama University Hospital Shuji Shimizu Department of Cardiovascular Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tsuyoshi Akiyama Department of Cardiac Physiology, National Cerebral and Cardiovascular Center Takanori Suezawa Department of Cardiovascular Surgery, Fukuyama City Hospital Yukio Kioka Department of Cardiovascular Surgery, Fukuyama City Hospital Yasuhiro Kotani Department of Cardiovascular Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences and Okayama University Hospital Toshiaki Shishido Department of Research Promotion and Management, National Cerebral and Cardiovascular Center Shingo Kasahara Department of Cardiovascular Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences and Okayama University Hospital It has been suggested that sodium-glucose cotransporter 2 (SGLT2) inhibitors have cardioprotective effects during myocardial ischemia/reperfusion (I/R) independent of glucose-lowering action. However, the effects of SGLT2 inhibitors on structural damage to cardiomyocytes in the ischemic region during I/R remain unknown. We applied a microdialysis technique to the heart of anesthetized rats and investigated the effects of an SGLT2 inhibitor, dapagliflozin, on myocardial interstitial myoglobin levels in the ischemic region during coronary occlusion followed by reperfusion. Dapagliflozin was administered systemically (40 mu g/body iv) or locally via a dialysis probe (100 mu M and 1 mM) 30 min before coronary occlusion. In the vehicle group, coronary occlusion increased the dialysate myoglobin concentration in the ischemic region. Reperfusion further increased the dialysate myoglobin concentration. Intravenous administration of dapagliflozin reduced dialysate myoglobin concentration during ischemia and at 0-15 min after reperfusion, but local administration (100 mu M and 1 mM) did not. Therefore, acute systemic administration of dapagliflozin prior to ischemia has cardioprotective effects on structural damage during I/R. No potential conflict of interest relevant to this article was reported.

American Diabetes Association Acta Medica Okayama 0012-1797 73 5 2024 GRP78 Contributes to the Beneficial Effects of SGLT2 Inhibitor on Proximal Tubular Cells in DKD 763 779 EN Atsuko Nakatsuka Division of Kidney, Diabetes and Endocrine Diseases, Okayama University Hospital Satoshi Yamaguchi Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Jun Wada Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University The beneficial effects of sodium–glucose cotransporter 2 (SGLT2) inhibitors on kidney function are well-known; however, their molecular mechanisms are not fully understood. We focused on 78-kDa glucose-regulated protein (GRP78) and its interaction with SGLT2 and integrin-β1 beyond the chaperone property of GRP78. In streptozotocin (STZ)-induced diabetic mouse kidneys, GRP78, SGLT2, and integrin-β1 increased in the plasma membrane fraction, while they were suppressed by canagliflozin. The altered subcellular localization of GRP78/integrin-β1 in STZ mice promoted epithelial mesenchymal transition (EMT) and fibrosis, which were mitigated by canagliflozin. High-glucose conditions reduced intracellular GRP78, increased its secretion, and caused EMT-like changes in cultured HK2 cells, which were again inhibited by canagliflozin. Urinary GRP78 increased in STZ mice, and in vitro experiments with recombinant GRP78 suggested that inflammation spread to surrounding tubular cells and that canagliflozin reversed this effect. Under normal glucose culture, canagliflozin maintained sarco/endoplasmic reticulum (ER) Ca2+-ATPase (SERCA) activity, promoted ER robustness, reduced ER stress response impairment, and protected proximal tubular cells. In conclusion, canagliflozin restored subcellular localization of GRP78, SGLT2, and integrin-β1 and inhibited EMT and fibrosis in DKD. In nondiabetic chronic kidney disease, canagliflozin promoted ER robustness by maintaining SERCA activity and preventing ER stress response failure, and it contributed to tubular protection. No potential conflict of interest relevant to this article was reported.

Pharmaceutical Society of Japan Acta Medica Okayama 0918-6158 45 11 2022 Investigation of the Expression of Serine Protease in Vibrio vulnificus 1596 1601 EN Tomoka Kawase Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Anusuya Debnath Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Tamaki Mizuno Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yui Miyake Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Vibrio vulnificus is a Gram-negative estuarine bacterium that causes infection in immuno-compromised patients, eels, and shrimp. V. vulnificus NCIMB2137, a metalloprotease-negative strain isolated from a diseased eel, produces a 45-kDa chymotrypsin-like alkaline serine protease known as VvsA. The gene encoding vvsA also includes another gene, vvsB with an unknown function; however, it is assumed to be an essential molecular chaperone for the maturation of VvsA. In the present study, we used an in vitro cell-free translation system to examine the maturation pathway of VvsA. We individually expressed the vvsA and vvsB genes and detected their mRNAs. However, the sample produced from vvsA did not exhibit protease activity. A sodium dodecyl sulfate (SDS) analysis detected the VvsB protein, but not the VvsA protein. A Western blotting analysis using a histidine (His)-tag at the amino terminus of proteins also showed no protein production by vvsA. These results suggested the translation, but not the transcription of vvsA. Factors derived from Escherichia coli were used in the in vitro cell-free translation system employed in the present study. The operon of the serine protease gene containing vvsA and vvsB was expressed in E. coli. Although serine proteases were produced, they were cleaved at different sites and no active mature forms were detected. These results indicate that the operon encoding vvsA and vvsB is a gene constructed to be specifically expressed in V. vulnificus. No potential conflict of interest relevant to this article was reported.

Springer Science and Business Media LLC Acta Medica Okayama 1936-0533 17 5 2023 Oxidative stress-related markers as prognostic factors for patients with primary sclerosing cholangitis in Japan 1215 1224 EN Atsushi Oyama Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Akinobu Takaki Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Takuya Adachi Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Nozomu Wada Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Yasuto Takeuchi Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Hideki Onishi Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Hidenori Shiraha Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Hiroyuki Okada Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Motoyuki Otsuka Department of Gastroenterology and Hepatology, Okayama University, Faculty of Medicine, Dentistry, and Pharmaceutical Sciences Background/purpose Primary sclerosing cholangitis (PSC) is a rare chronic liver disease. The mechanisms and prediction of PSC progression are unclear. Recent investigations have shown that general conditions, such as oxidative stress, affect the course of chronic diseases. We investigated the clinical course and oxidative stress-related condition of PSC to determine prognostic factors. Methods We recruited 58 patients with PSC (mean age; 37.4 years, mean observation period; 1382 days) who visited our department from 2003 to 2021. Clinical characteristics were investigated to define prognostic factors. Oxidative stress status was evaluated using two types of markers: an oxidative stress marker (serum reactive oxygen metabolite; dROM) and an antioxidant marker (serum OXY adsorbent test; OXY). Results The revised Mayo risk, Child–Pugh, model for end-stage liver disease-sodium (MELD-Na) scores or fibrosis-related FIB-4 index significantly predicted poor overall survival. High intestinal immunoglobulin A (IgA) levels predicted poor survival. Among patients with high and intermediate revised Mayo risk scores, those with physiologically high dROM levels showed better survival than those with lower dROM levels. In this population, dROM was negatively correlated with AST and IgA, which are both correlated with survival. Conclusions High and intermediate revised Mayo risk score group predicted a poor clinical course in PSC. Additionally, the Child–Pugh score, MELD-Na score, FIB-4 index, and serum IgA were significantly correlated with survival. In patients with high and intermediate revised Mayo risk scores, physiologically high oxidative stress status correlated with low IgA levels and a good prognosis. No potential conflict of interest relevant to this article was reported.

Ovid Technologies (Wolters Kluwer Health) Acta Medica Okayama 2641-7650 5 2 2023 Inhibition of Amino Acids Influx into Proximal Tubular Cells Improves Lysosome Function in Diabetes 182 194 EN Yuzuki Kano Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Satoshi Yamaguchi Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Koki Mise Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Chieko Kawakita Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yasuhiro Onishi Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Naoko Kurooka Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Ryosuke Sugawara Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Haya Hamed Hassan Albuayjan Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Atsuko Nakatsuka Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Jun Eguchi Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Jun Wada Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Background Inhibition of glucose influx into proximal tubular cells (PTCs) by sodium–glucose cotransporter 2 inhibitors revealed prominent therapeutic effects on diabetic kidney disease. Collectrin (CLTRN) serves as a chaperone for the trafficking of neutral amino acid (AA) transporters in the apical membranes of PTCs. We investigated the beneficial effects of reduced influx of AAs into PTCs in diabetes and obesity model of Cltrn－/y mice. Methods Cltrn+/y and Cltrn－/y mice at age 5 weeks were assigned to standard diet and streptozotocin and high-fat diet (STZ-HFD)–treated groups. Results At age 22–23 weeks, body weight and HbA1c levels significantly increased in STZ-HFD-Cltrn+/y compared with standard diet-Cltrn+/y; however, they were not altered in STZ-HFD-Cltrn－/y compared with STZ-HFD-Cltrn+/y. At age 20 weeks, urinary albumin creatinine ratio was significantly reduced in STZ-HFD-Cltrn－/y compared with STZ-HFD-Cltrn+/y. Under the treatments with STZ and HFD, the Cltrn gene deficiency caused significant increase in urinary concentration of AAs such as Gln, His, Gly, Thr, Tyr, Val, Trp, Phe, Ile, Leu, and Pro. In PTCs in STZ-HFD-Cltrn+/y, the enlarged lysosomes with diameter of 10 μm or more were associated with reduced autolysosomes, and the formation of giant lysosomes was prominently suppressed in STZ-HFD-Cltrn－/y. Phospho-mTOR and inactive form of phospho-transcription factor EB were reduced in STZ-HFD-Cltrn－/y compared with STZ-HFD-Cltrn+/y. Conclusions The reduction of AAs influx into PTCs inactivated mTOR, activated transcription factor EB, improved lysosome function, and ameliorated vacuolar formation of PTCs in STZ-HFD-Cltrn－/y mice. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 2040-1116 13 7 2022 Reduction in the magnitude of serum potassium elevation in combination therapy with esaxerenone (CS‐3150) and sodium–glucose cotransporter 2 inhibitor in patients with diabetic kidney disease: Subanalysis of two phase III studies 1190 1202 EN Kenichi Shikata Center for Innovative Clinical Medicine, Okayama University Hospital Sadayoshi Ito Division of Nephrology, Endocrinology and Vascular Medicine, Department of Medicine, Tohoku University School of Medicine Naoki Kashihara Department of Nephrology and Hypertension, Kawasaki Medical School Masaomi Nangaku Division of Nephrology and Endocrinology, Graduate School of Medicine, The University of Tokyo Takashi Wada Department of Nephrology and Laboratory Medicine, Kanazawa University Yasuyuki Okuda Department of Nephrology and Laboratory Medicine, Kanazawa University Tomoko Sawanobori Clinical Development Department, Daiichi Sankyo Co., Ltd. Kotaro Sugimoto Primary Medical Science Department, Daiichi Sankyo Co., Ltd. Aims/Introduction: We evaluated the effect of co-administration of esaxerenone and a sodium–glucose cotransporter 2 (SGLT2) inhibitor on the magnitude of serum potassium elevation in Japanese patients with diabetic kidney disease. Materials and Methods: We carried out a prespecified subanalysis of data from two phase III studies: a multicenter, randomized, double-blind, placebo-controlled trial in patients with type 2 diabetes and microalbuminuria (J308); and a multicenter, single-arm, open-label trial in patients with type 2 diabetes and macroalbuminuria (J309). Changes in serum potassium levels during the studies and other measures were evaluated according to SGLT2 inhibitor use. Results: In both studies, time-course changes in serum potassium levels, and incidence rates of serum potassium elevation were lower in patients with co-administration of SGLT2 inhibitor in both the placebo and esaxerenone groups than those without the inhibitor. In contrast, time-course changes and mean percentage changes from baseline in urinary albumin-to-creatinine ratio, the proportion of patients with albuminuria remission and time-course changes in blood pressure did not change with or without SGLT2 inhibitor, whereas the albumin-to-creatinine ratio and blood pressure were reduced with esaxerenone. The blood glucose-lowering effect of SGLT2 inhibitor was not affected by esaxerenone. Conclusions: In Japanese patients with type 2 diabetes and albuminuria treated with esaxerenone, concomitant use of SGLT2 inhibitor reduced the magnitude of serum potassium elevation without any change of its antihypertensive and albuminuria-suppressing effects. Co-administration of esaxerenone and SGLT2 inhibitor might be a beneficial treatment option for patients with diabetic kidney disease. 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.

Frontiers Media Acta Medica Okayama 2297-055X 10 2023 In vivo tracking transplanted cardiomyocytes derived from human induced pluripotent stem cells using nuclear medicine imaging 1261330 EN Yukihiro Saito Department of Cardiovascular Medicine, Okayama University Hospital Naoko Nose Molecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Toshihiro Iida Department of Cardiovascular Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Kaoru Akazawa Department of Cardiovascular Medicine, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takayuki Kanno Molecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuki Fujimoto Molecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takanori Sasaki Okayama Medical Innovation Center, Faculty of Medicine, Dentistry and Pharmaceutical Sciences Masaru Akehi Okayama Medical Innovation Center, Faculty of Medicine, Dentistry and Pharmaceutical Sciences Takahiro Higuchi Molecular Imaging Project of RECTOR Program, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Satoshi Akagi Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Masashi Yoshida Department of Chronic Kidney Disease and Cardiovascular Disease, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Toru Miyoshi Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroshi Ito Department of General Internal Medicine 3, Kawasaki Medical School Kazufumi Nakamura Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Introduction: Transplantation of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) is a promising treatment for heart failure. Information on long-term cell engraftment after transplantation is clinically important. However, clinically applicable evaluation methods have not yet been established. Methods: In this study, to noninvasively assess transplanted cell engraftment, human SLC5A5, which encodes a sodium/iodide symporter (NIS) that transports radioactive tracers such as 125I, 18F-tetrafluoroborate (TFB), and 99mTc-pertechnetate (99mTcO4－), was transduced into human induced pluripotent stem cells (iPSCs), and nuclear medicine imaging was used to track engrafted human iPSC-CMs. Results: To evaluate the pluripotency of NIS-expressing human iPSCs, they were subcutaneously transplanted into immunodeficient rats. Teratomas were detected by 99mTcO4－ single photon emission computed tomography (SPECT/CT) imaging. NIS expression and the uptake ability of 125I were maintained in purified human iPSC-CMs. NIS-expressing human iPSC-CMs transplanted into immunodeficient rats could be detected over time using 99mTcO4－ SPECT/CT imaging. Unexpectedly, NIS expression affected cell proliferation of human iPSCs and iPSC-derived cells. Discussion: Such functionally designed iPSC-CMs have potential clinical applications as a noninvasive method of grafted cell evaluation, but further studies are needed to determine the effects of NIS transduction on cellular characteristics and functions. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 1462-8902 24 8 2022 Rationale, design and baseline characteristics of the effect of canagliflozin in patients with type 2 diabetes and microalbuminuria in the Japanese population: The CANPIONE study 1429 1438 EN Satoshi Miyamoto Center for Innovative Clinical Medicine, Okayama University Hospital Hiddo J. L. Heerspink Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen Dick de Zeeuw Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen Masao Toyoda Division of Nephrology, Endocrinology and Metabolism, Department of Internal Medicine, Tokai University School of Medicine Daisuke Suzuki Suzuki Diadetes Clinic Takashi Hatanaka Department of Diabetes and Endocrinology, National Hospital Organization Fukuyama Medical Center Tohru Nakamura Diabetes Internal Medicine, Sumitomo Besshi Hospital Shinji Kamei Department of Diabetic Medicine, Kurashiki Central Hospital Satoshi Murao Department of Diabetes and Endocrinology, Takamatsu Hospital Kazuyuki Hida Department of Diabetology and Metabolism, National Hospital Organization Okayama Medical Center Shinichiro Ando Department of Internal Medicine Diabetic Center, Okayama City Hospital Hiroaki Akai Division of Diabetes and Metabolism, Faculty of Medicine, Tohoku Medical and Pharmaceutical University Yasushi Takahashi Department of Diabetes, Ochiai General Hospital Daisuke Koya Department of Diabetology and Endocrinology, Kanazawa Medical University Munehiro Kitada Department of Diabetology and Endocrinology, Kanazawa Medical University Hisashi Sugano Department of Diabetes and Endocrinology, Kochi Health Sciences Center Tomokazu Nunoue Nunoue Clinic Akihiko Nakamura Internal Medicine, Osafune Clinic, Setouchi Motofumi Sasaki Department of Diabetes and Endocrinology, Matsue City Hospital Tatsuaki Nakatou Diabetes Center, Okayama Saiseikai General Hospital Kei Fujimoto Division of Diabetes, Metabolism and Endocrinology, Department of Internal Medicine, The Jikei University Kashiwa Hospital Daiji Kawanami Department of Endocrinology and Diabetes Mellitus, Fukuoka University School of Medicine Takashi Wada Department of Nephrology and Laboratory Medicine, Graduate School of Medical Sciences, Kanazawa University Nobuyuki Miyatake Department of Hygiene, Faculty of Medicine, Kagawa University Michihiro Yoshida Center for Innovative Clinical Medicine, Okayama University Hospital Kenichi Shikata Center for Innovative Clinical Medicine, Okayama University Hospital the CANPIONE study Investigators Aim: To evaluate the effect of canagliflozin, a sodium-glucose co-transporter-2 (SGLT2) inhibitor, on albuminuria and the decline of estimated glomerular filtration rate (eGFR) in participants with type 2 diabetes and microalbuminuria. Methods: The CANPIONE study is a multicentre, randomized, parallel-group and open-labelled study consisting of a unique 24-week preintervention period, during which the rate of eGFR decline before intervention is estimated, followed by a 52-week intervention and a 4-week washout period. Participants with a geometric mean urinary albumin-to-creatinine ratio (UACR) of 50 and higher and less than 300 mg/g in two consecutive first-morning voids at two different time points, and an eGFR of 45 ml/min/1.73m2 or higher, are randomly assigned to receive canagliflozin 100 mg daily or to continue guideline-recommended treatment, except for SGLT2 inhibitors. The first primary outcome is the change in UACR, and the second primary outcome is the change in eGFR slope. Results: A total of 258 participants were screened and 98 were randomized at 21 sites in Japan from August 2018 to May 2021. The mean baseline age was 61.4 years and 25.8% were female. The mean HbA1c was 7.9%, mean eGFR was 74.1 ml/min/1.73m2 and median UACR was 104.2 mg/g. Conclusions: The CANPIONE study will determine whether the SGLT2 inhibitor canagliflozin can reduce albuminuria and slow eGFR decline in participants with type 2 diabetes and microalbuminuria. No potential conflict of interest relevant to this article was reported.

Elsevier BV Acta Medica Okayama 0021-9673 1706 2023 Separation and fractionation of glutamic acid and histidine via origami isoelectric focusing 464247 EN Kaewta Danchana Department of Chemistry, Okayama University Nayu Yamashita Department of Chemistry, Okayama University Mika I. Umeda Department of Chemistry, Okayama University Takashi Kaneta Department of Chemistry, Okayama University We demonstrated the fractionation of two amino acids, glutamic acid and histidine, separated via isoelectric focusing (IEF) on filter paper folded and stacked in an origami fashion. Channels for electrophoresis were fabricated as circular zones acquired via wax printing onto the filter paper. An ampholyte solution with amphiphilic samples was deposited on all the circle zones, which was followed by folding to form the electrophoresis channels. IEF was achieved by applying an electrical potential between the anodic and cathodic chambers filled with phosphoric acid and sodium hydroxide solutions, respectively. A pH gradient was formed using either a wide-range ampholyte with a pH of 3 to 10 or a narrow-range version with a pH of 5 to 8, which was confirmed by adding pH indicators to each layer. The origami IEF was used to separate the amino acids, glutamic acid and histidine, by mixing with the ampholytes, which were deposited on the layers. The components in each layer were extracted with water and measured by high-performance liquid chromatography using pre-column derivatization with dansyl chloride. The results indicated that the focus for glutamic acid and that for histidine were at different layers, according to their isoelectric points. The origami isoelectric focusing achieved the fractionation of amino acids in less than 3 min using voltage as low as 30 V. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 77 4 2023 Changes in TRPV1 Receptor, CGRP, and BDNF Expression in Rat Dorsal Root Ganglion with Resiniferatoxin-Induced Neuropathic Pain: Modulation by Pulsed Radiofrequency Applied to the Sciatic Nerve 359 364 EN Tomohiro Koshida Department of Anesthesiology and Pain Clinic, Faculty of Medicine, University of Miyazaki Toyoaki Maruta Department of Anesthesiology and Pain Clinic, Faculty of Medicine, University of Miyazaki Nobuhiko Tanaka Tanaka homecare clinic Kotaro Hidaka Department of Anesthesiology and Pain Clinic, Faculty of Medicine, University of Miyazaki Mio Kurogi Department of Anesthesiology and Pain Clinic, Faculty of Medicine, University of Miyazaki Takayuki Nemoto Department of Pharmacology, Faculty of Medicine, Fukuoka University Toshihiko Yanagita Department of Clinical Pharmacology, School of Nursing, Faculty of Medicine, University of Miyazaki Ryu Takeya Department of Pharmacology, Faculty of Medicine, University of Miyazaki Isao Tsuneyoshi Department of Anesthesiology and Pain Clinic, Faculty of Medicine, University of Miyazaki Original Article 10.18926/AMO/65741 Pulsed radiofrequency (PRF) is a safe method of treating neuropathic pain by generating intermittent electric fields at the needle tip. Resiniferatoxin (RTX) is an ultrapotent agonist of transient receptor potential vanilloid subtype-1 (TRPV1) receptors. We investigated the mechanism of PRF using a rat model of RTX-induced neuropathic pain. After administering RTX intraperitoneally, PRF was applied to the right sciatic nerve. We observed the changes in TRPV1, calcitonin gene-related peptide (CGRP), and brain-derived neurotrophic factor (BDNF) in the dorsal root ganglia by western blotting. Expressions of TRPV1 and CGRP were significantly lower in the contralateral (RTX-treated, PRF-untreated) tissue than in control rats (p<0.0001 and p<0.0001, respectively) and the ipsilateral tissues (p<0.0001 and p<0.0001, respectively). BDNF levels were significantly higher in the contralateral tissues than in the control rats (p<0.0001) and the ipsilateral tissues (p<0.0001). These results suggest that, while TRPV1 and CGRP are decreased by RTX-induced neuronal damage, increased BDNF levels result in pain development. PRF may promote recovery from neuronal damage with concomitant restoration of TRPV1 and CGRP, and exert its analgesic effect by reversing BDNF increase. Further research is required to understand the role of TRPV1 and CGRP restoration in improving mechanical allodynia. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 1347-9032 114 10 2023 Combination therapy with hydrogen peroxide and irradiation promotes an abscopal effect in mouse models 3848 3856 EN Naoya Kemmotsu Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Li Zhu Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Joji Nagasaki Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yoshihiro Otani Department of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Youki Ueda Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiromichi Dansako Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yue Fang Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University Isao Date Department of Neurological Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yosuke Togashi Department of Tumor Microenvironment, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hydrogen peroxide (H2O2) induces oxidative stress and cytotoxicity, and can be used for treating cancers in combination with radiotherapy. A product comprising H2O2 and sodium hyaluronate has been developed as a radiosensitizer. However, the effects of H2O2 on antitumor immunity remain unclear. To investigate the effects of H2O2, especially the abscopal effect when combined with radiotherapy (RT), we implanted murine tumor cells simultaneously in two locations in mouse models: the hind limb and back. H2O2 mixed with sodium hyaluronate was injected intratumorally, followed by irradiation only at the hind limb lesion. No treatment was administered to the back lesion. The H2O2/RT combination significantly reduced tumor growth at the noninjected/nonirradiated site in the back lesion, whereas H2O2 or RT individually did not reduce tumor growth. Flow cytometric analyses of the tumor-draining lymph nodes in the injected/irradiated areas showed that the number of dendritic cells increased significantly with maturation in the H2O2/RT combination group. In addition, analyses of tumor-infiltrating lymphocytes showed that the number of CD8+ (cluster of differentiation 8) T cells and the frequency of IFN-γ+ (interferon gamma) CD8+ T cells were higher in the noninjected/nonirradiated tumors in the H2O2/RT group compared to those in the other groups. PD-1 (programmed death receptor 1) blockade further increased the antitumor effect against noninjected/nonirradiated tumors in the H2O2/RT group. Intratumoral injection of H2O2 combined with RT therefore induces an abscopal effect by activating antitumor immunity, which can be further enhanced by PD-1 blockade. These findings promote the development of H2O2/RT therapy combined with cancer immunotherapies, even for advanced cancers. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2072-6643 15 13 2023 Adrenomedullin Enhances Mouse Gustatory Nerve Responses to Sugars via T1R-Independent Sweet Taste Pathway 2941 EN Shusuke Iwata Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University Ryusuke Yoshida Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Shingo Takai Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University Keisuke Sanematsu Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University Noriatsu Shigemura Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University Yuzo Ninomiya Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University On the tongue, the T1R-independent pathway (comprising glucose transporters, including sodium-glucose cotransporter (SGLT1) and the K-ATP channel) detects only sugars, whereas the T1R-dependent (T1R2/T1R3) pathway can broadly sense various sweeteners. Cephalic-phase insulin release, a rapid release of insulin induced by sensory signals in the head after food-related stimuli, reportedly depends on the T1R-independent pathway, and the competitive sweet taste modulators leptin and endocannabinoids may function on these two different sweet taste pathways independently, suggesting independent roles of two oral sugar-detecting pathways in food intake. Here, we examined the effect of adrenomedullin (ADM), a multifunctional regulatory peptide, on sugar sensing in mice since it affects the expression of SGLT1 in rat enterocytes. We found that ADM receptor components were expressed in T1R3-positive taste cells. Analyses of chorda tympani (CT) nerve responses revealed that ADM enhanced responses to sugars but not to artificial sweeteners and other tastants. Moreover, ADM increased the apical uptake of a fluorescent D-glucose derivative into taste cells and SGLT1 mRNA expression in taste buds. These results suggest that the T1R-independent sweet taste pathway in mouse taste cells is a peripheral target of ADM, and the specific enhancement of gustatory nerve responses to sugars by ADM may contribute to caloric sensing and food intake. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1999-4923 15 1 2022 Formation of a Stable Co-Amorphous System for a Brick Dust Molecule by Utilizing Sodium Taurocholate with High Glass Transition Temperature 84 EN Shohei Aikawa Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Okayama University Hironori Tanaka Formulation Research Department, Formulation R&D Laboratory, Shionogi & Co., Ltd. Hiroshi Ueda Bioanalytical, Analysis and Evaluation Laboratory, Shionogi & Co., Ltd. Masato Maruyama Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Okayama University Kazutaka Higaki Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Okayama University Brick dust molecules are usually poorly soluble in water and lipoidal components, making it difficult to formulate them in dosage forms that provide efficient pharmacological effects. A co-amorphous system is an effective strategy to resolve these issues. However, their glass transition temperatures (Tg) are relatively lower than those of polymeric amorphous solid dispersions, suggesting the instability of the co-amorphous system. This study aimed to formulate a stable co-amorphous system for brick dust molecules by utilizing sodium taurocholate (NaTC) with a higher Tg. A novel neuropeptide Y-5 receptor antagonist (AntiY(5)R) and NaTC with Tg of 155 degrees C were used as the brick dust model and coformer, respectively. Ball milling formed a co-amorphous system for AntiY(5)R and NaTC (AntiY(5)R-NaTC) at various molar ratios. Deviation from the theoretical Tg value and peak shifts in Fourier-transform infrared spectroscopy indicated intermolecular interactions between AntiY(5)R and NaTC. AntiY(5)R-NaTC at equal molar ratios resulting in an 8.5-fold increase in AntiY(5)R solubility over its crystalline form. The co-amorphous system remained amorphous for 1 month at 25 degrees C and 40 degrees C. These results suggest that the co-amorphous system formed by utilizing NaTC as a coformer could stably maintain the amorphous state and enhance the solubility of brick dust molecules. No potential conflict of interest relevant to this article was reported.

Hindawi Ltd. Acta Medica Okayama 1536-0121 2022 2022 In Vivo Functional Assessment of Sodium-Glucose Cotransporters (SGLTs) Using [F-18]Me4FDG PET in Rats 4635171 EN Yohji Matsusaka Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital of Würzburg Xinyu Chen Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital of Würzburg Paula Arias-Loza Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital of Würzburg Rudolf A. Werner Department of Nuclear Medicine and Comprehensive Heart Failure Center, University Hospital of Würzburg Naoko Nose Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takanori Sasaki Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Steven P. Rowe Division of Nuclear Medicine and Molecular Imaging, The Russell H Morgan Department of Radiology and Radiological Sciences, Johns Hopkins School of Medicine Martin G. Pomper Division of Nuclear Medicine and Molecular Imaging, The Russell H Morgan Department of Radiology and Radiological Sciences, Johns Hopkins School of Medicine Constantin Lapa Nuclear Medicine, Faculty of Medicine, University of Augsburg Takahiro Higuchi Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Background. Mediating glucose absorption in the small intestine and renal clearance, sodium glucose cotransporters (SGLTs) have emerged as an attractive therapeutic target in diabetic patients. A substantial fraction of patients, however, only achieve inadequate glycemic control. Thus, we aimed to assess the potential of the SGLT-targeting PET radiotracer alpha-methyl-4-deoxy-4-[F-18]fluoro-D-glucopyranoside ([F-18]Me4FDG) as a noninvasive intestinal and renal biomarker of SGLT-mediated glucose transport. Methods. We investigated healthy rats using a dedicated small animal PET system. Dynamic imaging was conducted after administration of the reference radiotracer 2-deoxy-2-[F-18]fluoro-D-glucose ([F-18]FDG), or the SGLT-targeting agent, [F-18]Me4FDG either directly into the digestive tract (for assessing intestinal absorption) or via the tail vein (for evaluating kidney excretion). To confirm the specificity of [F-18]Me4FDG and responsiveness to treatment, a subset of animals was also pretreated with the SGLT inhibitor phlorizin. In this regard, an intraintestinal route of administration was used to assess tracer absorption in the digestive tract, while for renal assessment, phlorizin was injected intravenously (IV). Results. Serving as reference, intestinal administration of [F-18]FDG led to slow absorption with retention of 89.2 +/- 3.5% of administered radioactivity at 15 min. [F-18]Me4FDG, however, was rapidly absorbed into the blood and cleared from the intestine within 15 min, leading to markedly lower tracer retention of 18.5 +/- 1.2% (P < 0.0001). Intraintestinal phlorizin led to marked increase of [F-18]Me4FDG uptake (15 min, 99.9 +/- 4.7%; P < 0.0001 vs. untreated controls), supporting the notion that this PET agent can measure adequate SGLT inhibition in the digestive tract. In the kidneys, radiotracer was also sensitive to SGLT inhibition. After IV injection, [F-18]Me4FDG reabsorption in the renal cortex was significantly suppressed by phlorizin when compared to untreated animals (%ID/g at 60 min, 0.42 +/- 0.10 vs. untreated controls, 1.20 +/- 0.03; P < 0.0001). Conclusion. As a noninvasive read-out of the concurrent SGLT expression in both the digestive tract and the renal cortex, [F-18]Me4FDG PET may serve as a surrogate marker for treatment response to SGLT inhibition. As such, [F-18]Me4FDG may enable improvement in glycemic control in diabetes by PET-based monitoring strategies. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 6 2022 Graphene Oxide-based Endodontic Sealer: An in Vitro Study 715 721 EN Mohammed Zahedul Islam Nizami Department of Pathophysiology – Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Melahat Gorduysus Department of Pathophysiology – Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuki Shinoda-Ito Department of Pathophysiology – Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Tadashi Yamamoto Department of Pathophysiology – Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yuta Nishina Research Core for Interdisciplinary Sciences, Okayama University Shogo Takashiba Research Core for Interdisciplinary Sciences, Okayama University Zulema Arias Department of Pathophysiology – Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Original Article 10.18926/AMO/64122 The failure of endodontic treatment is directly associated with microbial infection in the root canal or periapical areas. An endodontic sealer that is both bactericidal and biocompatible is essential for the success of root canal treatments. This is one of the vital issues yet to be solved in clinical dental practice. This in vitro study assessed the effectiveness of graphene oxide (GO) composites GO-CaF2 and GO-Ag-CaF2 as endodontic sealer materials. Dentin slices were coated with either the GO-based composites or commonly used root canal sealers (non-eugenol zinc oxide sealer). The coated slices were treated in 0.9% NaCl, phosphate-buffered saline (PBS), and simulated body fluid (SBF) at 37˚C for 24 hours to compare their sealing effect on the dentin surface. In addition, the radiopacity of these composites was examined to assess whether they complied with the requirements of a sealer for good radiographic visualization. Scanning electron microscopy showed the significant sealing capability of the composites as coating materials. Radiographic images confirmed their radiopacity. Mineral deposition indicated their bioactivity, especially of GO-Ag-CaF2, and thus it is potential for regenerative application. They were both previously shown to be bactericidal to oral microbes and cytocompatible with host cells. With such a unique assemblage of critical properties, these GO-based composites show promise as endodontic sealers for protection against reinfection in root canal treatment and enhanced success in endodontic treatment overall. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 76 6 2022 The Impact of Tofogliflozin on Physiological and Hormonal Function, Serum Electrolytes, and Cardiac Diastolic Function in Elderly Japanese Patients with Type 2 Diabetes Mellitus 705 713 EN Toshihiro Higashikawa Kanazawa Medical University Himi Municipal Hospital Tomohiko Ito Kanazawa Medical University Himi Municipal Hospital Takurou Mizuno Kanazawa Medical University Himi Municipal Hospital Keiichiro Ishigami Kanazawa Medical University Himi Municipal Hospital Kengo Kuroki Kanazawa Medical University Himi Municipal Hospital Naoto Maekawa Kanazawa Medical University Himi Municipal Hospital Daisuke Usuda Kanazawa Medical University Himi Municipal Hospital Toshihide Izumida Kanazawa Medical University Himi Municipal Hospital Shinya Yamada Kanazawa Medical University Himi Municipal Hospital Ryusho Sangen Kanazawa Medical University Himi Municipal Hospital Kazu Hamada Kanazawa Medical University Himi Municipal Hospital Jun Kiyosawa Kanazawa Medical University Himi Municipal Hospital Atsushi Saito Kanazawa Medical University Himi Municipal Hospital Masaharu Iguchi Kanazawa Medical University Himi Municipal Hospital Yuji Kasamaki Kanazawa Medical University Himi Municipal Hospital Takeshi Nakahashi Kanazawa Medical University Himi Municipal Hospital Akihiro Fukuda Kanazawa Medical University Himi Municipal Hospital Hitoshi Saito Kanazawa Medical University Himi Municipal Hospital Tsugiyasu Kanda Kanazawa Medical University Himi Municipal Hospital Masashi Okuro Department of Geriatric Medicine, Kanazawa Medical University Original Article 10.18926/AMO/64121 The sodium glucose transporter 2 (SGLT2) inhibitor tofogliflozin is a glucose-lowering drug that causes the excretion of surplus glucose by inhibiting SGLT2. Because of tofogliflozin’s osmotic diuresis mechanism, patients’ serum electrolytes, body fluid levels, and cardiac function must be monitored. We retrospectively analyzed the cases of 64 elderly Japanese patients with type 2 diabetes mellitus (T2DM) who received tofogliflozin for 3 months. Their HbA1c, serum electrolytes (sodium, potassium, chloride), hematocrit, brain natriuretic peptide (cardiac volume load marker) and renin and aldosterone (RAA; an index of regulatory hormones involved in body fluid retention) were continuously monitored during the investigation period. Renal function and cardiac function (by echocardiography) were assessed throughout the period. HbA1c significantly decreased (β1=－0.341, p<0.0001, linear regression analysis [LRA]). Most of the hormonal, electrolyte, and physiological parameters were maintained throughout the study period. In these circumstances, E/e’ tended to decrease (β1=－0.382, p=0.13, LRA). Compared to the baseline, E/e’ was significantly decreased at 1 and 3 months (p<0.01, p<0.05). In the higher E/e’ group (E/e’≥10, n=34), E/e’ decreased significantly (β1=－0.63, p<0.05, LRA). ΔE/e’ was correlated with body-weight change during treatment (r=0.64, p<0.01). The 3-month tofogliflozin treatment improved glycemic control and diastolic function represented by E/e’ in T2DM patients, without affecting serum electrolytes, renal function, or RAA. No negative impacts on the patients were observed. Three-month tofogliflozin treatment lowered glucose and improved cardiac diastolic function. No potential conflict of interest relevant to this article was reported.

Nature Portfolio Acta Medica Okayama 2045-2322 12 1 2022 Effects of luseogliflozin and voglibose on high-risk lipid profiles and inflammatory markers in diabetes patients with heart failure 15449 EN Kentaro Ejiri Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toru Miyoshi Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hajime Kihara Department of Internal Medicine, Kihara Cardiovascular Clinic Yoshiki Hata Department of Cardiology, Minamino Cardiovascular Hospital Toshihiko Nagano Department of Internal Medicine, Iwasa Hospital Atsushi Takaishi Department of Cardiology, Mitoyo General Hospital Hironobu Toda Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Seiji Namba Department of Cardiology, Okayama Rosai Hospital Yoichi Nakamura Department of Cardiovascular Medicine, Specifed Clinic of Soyokaze Cardiovascular Medicine and Diabetes Care Satoshi Akagi Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Satoru Sakuragi Department of Cardiovascular Medicine, Iwakuni Clinical Center Taro Minagawa Department of Internal Medicine, Minagawa Cardiovascular Clinic Yusuke Kawai Department of Cardiovascular Medicine, Okayama City Hospital Nobuhiro Nishii Department of Internal Medicine, Yoshinaga Hospital Soichiro Fuke Department of Cardiovascular Medicine, Japanese Red Cross Okayama Hospital Masaki Yoshikawa Department of Cardiology, Fukuyama City Hospital Kazufumi Nakamura Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroshi Ito Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences The MUSCAT-HF Study Investigators Sodium-glucose cotransporter 2 inhibitors could reduce cardiovascular events in patients with heart failure irrespective of diabetes status. In this prespecified sub-analysis of randomised-controlled trial, we investigated the efficacy of luseogliflozin (2.5 mg daily), a sodium-glucose cotransporter 2 inhibitor, with that of voglibose (0.6 mg daily), an alpha-glucosidase inhibitor, on high-risk lipid profile and inflammatory markers in patients with type-2 diabetes and heart failure. Among the 157 patients studied, there were no significant differences in the mean malondialdehyde LDL or small-dense LDL cholesterol levels between the luseogliflozin and voglibose groups (percent change: 0.2% vs. - 0.6%, p = 0.93; - 1.7% vs. - 8.6%, p= 0.21) after 12 weeks in comparison to levels at the baseline. No significant difference was observed between the two groups in the adiponectin and high-sensitivity C-reactive protein levels after 12 weeks compared to the baseline levels (percent change, - 1.6% vs. - 4.0% and 22.5% vs. 10.0%; p = 0.52 and p = 0.55, respectively). In conclusion, in patients with type-2 diabetes and heart failure, compared to voglibose, luseogliflozin did not significantly improve the high-risk lipoprotein profile including malondialdehyde LDL and small-dense LDL cholesterol or the levels of inflammatory markers, including adiponectin and high-sensitivity C-reactive protein. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 23 7 2022 Pathophysiology and Treatment of Diabetic Cardiomyopathy and Heart Failure in Patients with Diabetes Mellitus 3587 EN Kazufumi Nakamura Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Toru Miyoshi Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Masashi Yoshida Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Satoshi Akagi Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yukihiro Saito Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Kentaro Ejiri Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Naoaki Matsuo Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Keishi Ichikawa Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Keiichiro Iwasaki Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Takanori Naito Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yusuke Namba Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Masatoki Yoshida Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroki Sugiyama Department of Internal Medicine, Okayama Saiseikai General Hospital Hiroshi Ito Department of Cardiovascular Medicine, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University There is a close relationship between diabetes mellitus and heart failure, and diabetes is an independent risk factor for heart failure. Diabetes and heart failure are linked by not only the complication of ischemic heart disease, but also by metabolic disorders such as glucose toxicity and lipotoxicity based on insulin resistance. Cardiac dysfunction in the absence of coronary artery disease, hypertension, and valvular disease is called diabetic cardiomyopathy. Diabetes-induced hyperglycemia and hyperinsulinemia lead to capillary damage, myocardial fibrosis, and myocardial hypertrophy with mitochondrial dysfunction. Lipotoxicity with extensive fat deposits or lipid droplets is observed on cardiomyocytes. Furthermore, increased oxidative stress and inflammation cause cardiac fibrosis and hypertrophy. Treatment with a sodium glucose cotransporter 2 (SGLT2) inhibitor is currently one of the most effective treatments for heart failure associated with diabetes. However, an effective treatment for lipotoxicity of the myocardium has not yet been established, and the establishment of an effective treatment is needed in the future. This review provides an overview of heart failure in diabetic patients for the clinical practice of clinicians. No potential conflict of interest relevant to this article was reported.

Wiley Periodicals, Inc Acta Medica Okayama 2055-5822 2021 Effects of luseogliflozin on estimated plasma volume in patients with heart failure with preserved ejection fraction EN Mitsutaka Nakashima Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toru Miyoshi Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kentaro Ejiri Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hajime Kihara Department of Internal Medicine, Kihara Cardiovascular Clinic Yoshiki Hata Department of Cardiology, Minamino Cardiovascular Hospital Toshihiko Nagano Department of Internal Medicine, Iwasa Hospital, Atsushi Takaishi Department of Cardiology, Mitoyo General Hospital Hironobu Toda Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Seiji Nanba Department of Cardiology, Okayama Rosai Hospital Yoichi Nakamura Department of Cardiovascular Medicine, Specified Clinic of Soyokaze CardioVascular Medicine and Diabetes Care, Matsuyama Satoshi Akagi Department of Internal Medicine, Akaiwa Medical Association Hospital Satoru Sakuragi Department of Cardiovascular Medicine, Iwakuni Clinical Center Taro Minagawa Department of Internal Medicine, Minagawa Cardiovascular Clinic Yusuke Kawai Department of Cardiovascular Medicine, Okayama City Hospital Nobuhiro Nishii Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Soichiro Fuke Department of Cardiovascular Medicine, Japanese Red Cross Okayama Hospital Masaki Yoshikawa Department of Cardiology, Fukuyama City Hospital Kazufumi Nakamura Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroshi Ito Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences MUSCAT-HF Study Investigators Aims Sodium glucose co-transporter 2 inhibitors have diuretic effects in both patients with glycosuria and with natriuresis. We sought to assess the effect of luseogliflozin on estimated plasma volume (ePV) in patients with type 2 diabetes and heart failure with preserved ejection fraction (HFpEF). Methods and results This study was a post-hoc analysis of the MUSCAT-HF trial (UMIN000018395), a multicentre, prospective, open-label, randomized controlled trial that assessed the effect of 12 weeks of luseogliflozin (2.5 mg, once daily, n = 83) as compared with voglibose (0.2 mg, three times daily, n = 82) on the reduction in brain natriuretic peptide (BNP) in patients with type 2 diabetes and HFpEF. The analysis compared the change in ePV calculated by the Straus formula from baseline to Weeks 4, 12, and 24, using a mixed-effects model for repeated measures. We also estimated the association between changes in ePV and changes in other clinical parameters, including BNP levels. Luseogliflozin significantly reduced ePV as compared to voglibose at Week 4 {adjusted mean group-difference -6.43% [95% confidence interval (CI): -9.11 to -3.74]}, at Week 12 [-8.73% (95%CI: -11.40 to -6.05)], and at Week 24 [-11.02% (95%CI: -13.71 to -8.33)]. The effect of luseogliflozin on these parameters was mostly consistent across various patient clinical characteristics. The change in ePV at Week 12 was significantly associated with log-transformed BNP (r = 0.197, P = 0.015) and left atrial volume index (r = 0.283, P = 0.019). Conclusions Luseogliflozin significantly reduced ePV in patients with type 2 diabetes and HFpEF, as compared with voglibose. The reduction of intravascular volume by luseogliflozin may provide clinical benefits to patients with type 2 diabetes and HFpEF. No potential conflict of interest relevant to this article was reported.

American Physical Society (APS) Acta Medica Okayama 2469-9950 104 14 2021 Spin-gap formation due to spin-Peierls instability in π-orbital-ordered NaO2 L140402 EN Mizuki Miyajima Department of Physics, Okayama University Fahmi Astuti Advanced Meson Science Laboratory, RIKEN Nishina Center Takahito Fukuda Department of Physics, Okayama University Masashi Kodani Department of Physics, Okayama University Shinsuke Iida Institute for Solid State Physics, University of Tokyo Shinichiro Asai Institute for Solid State Physics, University of Tokyo Akira Matsuo Institute for Solid State Physics, University of Tokyo Takatsugu Masuda Institute for Solid State Physics, University of Tokyo Koichi Kindo Institute for Solid State Physics, University of Tokyo Takumi Hasegawa Graduate School of Advanced Science and Engineering, Hiroshima University Tatsuo C. Kobayashi Department of Physics, Okayama University Takehito Nakano Institute of Quantum Beam Science, Ibaraki University Isao Watanabe Advanced Meson Science Laboratory, RIKEN Nishina Center Takashi Kambe Department of Physics, Okayama University We have investigated the low-temperature magnetism of sodium superoxide (NaO2), in which spin, orbital, and lattice degrees of freedom are closely entangled. The magnetic susceptibility shows anomalies at T1 = 220 K and T2 = 190 K, which correspond well to the structural phase transition temperatures, and a sudden decrease below T3 = 34 K. At 4.2 K, the magnetization shows a clear stepwise anomaly around 30 T with a large hysteresis. In addition, the muon spin relaxation experiments indicate no magnetic phase transition down to T = 0.3 K. The inelastic neutron scattering spectrum exhibits magnetic excitation with a finite energy gap. These results confirm that the ground state of NaO2 is a spin-singlet state. To understand this ground state in NaO2, we performed Raman scattering experiments. All the Raman-active libration modes expected for the marcasite phase below T2 are observed. Furthermore, we find that several new peaks appear below T3. This directly evidences the low crystal symmetry, namely, the presence of the phase transition at T3.We conclude that the singlet ground state of NaO2 is due to the spin-Peierls instability. 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.

BMC Acta Medica Okayama 1471-2369 22 1 2021 Overexpression of SGLT2 in the kidney of a P. gingivalis LPS-induced diabetic nephropathy mouse model 287 EN Koichiro Kajiwara Department of Oral Growth & Development, Fukuoka Dental College Yoshihiko Sawa Department of Oral Function & Anatomy, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Background: The overexpression of sodium-glucose cotransporter 2 (SGLT2) in diabetic kidneys has been reported. It has also been established that the diabetic glomerular endothelium expresses the toll-like receptors TLR2 and TLR4. The present study aims to examine the renal SGLT2 induction by the TLR2/4 ligand Porphyromonas (P.) gingivalis lipopolysaccharide (Pg-LPS) in mouse diabetic nephropathy. Methods: Immunohistochemical study and tissue RT-PCR analyses were performed on mouse kidneys in streptozotocin (STZ)-induced diabetic ICR mice (STZ-ICR), in healthy ICR mice administered Pg-LPS (LPS-ICR), and in diabetic ICR mouse kidneys with Pg-LPS-induced nephropathy (LPS-STZ). Results: In the quantitative analysis of blood sugar levels, the mean time to reach 600 mg/dl was shorter in the LPS-STZ than in the STZ-ICR kidneys. The rise in blood glucose levels was significantly steeper in the LPS-STZ than in the STZ-ICR kidneys. According to these data the LPS-STZ model suggests a marked glucose intolerance. The expression of SGLT2 was significantly stronger in the whole of the renal parenchyma of the LPS-STZ than in the LPS-ICR or in the STZ-ICR. The expression of SGLT2 was observed both in the renal tubules and around the renal tubules, and in the glomeruli of the LPS-STZ kidneys. In the analysis by tissue real-time PCR and cell ELISA, the expression of the SGLT2 gene and protein was significantly stronger in the LPS-STZ than in the LPS-ICR or in the STZ-ICR. There were no differences in the renal SGLT2 production in the LPS-ICR and the STZ-ICR kidneys. Conclusions: Abnormally high renal expression of SGLT2 occurs in diabetic kidneys with P. gingivalis LPS. Periodontitis may be an exacerbating factor in diabetic nephropathy as well as in diabetes. No potential conflict of interest relevant to this article was reported.

American Chemical Society (ACS) Acta Medica Okayama 1520-6106 125 23 2021 Ion Size Dependences of the Salting-Out Effect: Reversed Order of Sodium and Lithium Ions 6296 6305 EN Hiroyuki Katsuto Research Institute for Interdisciplinary Science, Okayama University Ryuichi Okamoto Research Institute for Interdisciplinary Science, Okayama University Tomonari Sumi Research Institute for Interdisciplinary Science, Okayama University Kenichiro Koga Research Institute for Interdisciplinary Science, Okayama University A general trend of the salting-out effect on hydrophobic solutes in aqueous solution is that the smaller the size of a dissolved ion, the larger the effect of reducing the solubility of a hydrophobe. An exception is that Li+, the smallest in alkali metal ions, has a notably weaker effect than Na+. To understand the reversed order in the cation series, we performed molecular dynamics simulations of aqueous solutions of salt ions and calculated the Setschenow coefficient of methane with the ionic radius of either a cation or an anion varied in a wide range. It is confirmed that the Setschenow coefficient is correlated with the packing fraction of salt solution, as observed in earlier studies, and also correlated with the partial molar volume of an ion. Analyses of correlation function integrals, packing fractions of solvation spheres, and orientations of water molecules surrounding an ion reveal the key differences in microscopic properties between the cation and anion series, which give rise to the reversed order in the cation series of the partial molar volumes of ions and ultimately that of the Setschenow coefficients. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2311-5637 7 2 2021 Anti-Inflammatory Effect on Colitis and Modulation of Microbiota by Fermented Plant Extract Supplementation 55 EN Manabu Sugimoto Institute of Plant Science and Resources, Okayama University Toshiro Watanabe Department of Food and Nutrition, Sonoda Women’s University Motoko Takaoka Department of Biosphere Sciences, Kobe College Kyoko Suzuki Department of Biosphere Sciences, Kobe College Tadatoshi Murakami Functional Food Creation Research Institute Co., Ltd. Nobutada Murakami Functional Food Creation Research Institute Co., Ltd. Shoichi Sumikawa Functional Food Creation Research Institute Co., Ltd. Although results of recent studies suggest that fermented foods strongly affect the gut microbiota composition and that they relieve inflammatory bowel disease symptoms, some reports have described that fermented foods increase some inflammation markers based on differences in fermented food materials. This study evaluated the effects of fermented plant extract (FPE) on dextran sulfate sodium (DSS)-induced colitis in mice and the effects on fecal microbiota composition in humans. Mice fed 5% FPE with 3% DSS (FPE group) showed no body weight loss, atrophy of colonic length, or bloody stool, similar to mice fed a basal diet (negative group), whereas mice fed 3% DSS (positive group) exhibited those effects. Concentrations of inflammation markers IL-6 and TNF-alpha were not significantly different between FPE and negative groups; however, those concentrations became higher in the positive group. 16S ribosomal RNA gene sequencing was used to characterize fecal microbiota in healthy women before and after 3-month FPE supplementation. The FPE supplementation induced increases in Firmicutes phyla and in Clostridiales order, which play a central role in inflammation suppression. These results suggest that FPE enhances Clostridiales growth in the gut and that it has an anti-inflammatory effect. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 22 11 2021 Management of Cirrhotic Ascites under the Add-on Administration of Tolvaptan 5582 EN Takuya Adachi Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yasuto Takeuchi Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Akinobu Takaki Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Atsushi Oyama Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Nozomu Wada Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hideki Onishi Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hidenori Shiraha Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroyuki Okada Department of Gastroenterology and Hepatology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Tolvaptan is a recently available diuretic that blocks arginine vasopressin receptor 2 in the renal collecting duct. Its diuretic mechanism involves selective water reabsorption by affecting the water reabsorption receptor aquaporin 2. Given that liver cirrhosis patients exhibit hyponatremia due to their pseudo-aldosteronism and usage of natriuretic agents, a sodium maintaining agent, such as tolvaptan, is physiologically preferable. However, large scale studies indicating the patients for whom this would be effective and describing management under its use have been insufficient. The appropriate management of cirrhosis patients treated with tolvaptan should be investigated. In the present review, we collected articles investigating the effectiveness of tolvaptan and factors associated with survival and summarized their management reports. Earlier administration of tolvaptan before increasing the doses of natriuretic agents is recommended because this may preserve effective arterial blood volume. No potential conflict of interest relevant to this article was reported.

Nature Research Acta Medica Okayama 2399-3669 4 1 2021 Halogen-sodium exchange enables efficient access to organosodium compounds 76 EN Sobi Asako Division of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University Ikko Takahashi RIKEN Center for Sustainable Resource Science Hirotaka Nakajima Division of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University Laurean Ilies RIKEN Center for Sustainable Resource Science Kazuhiko Takai Division of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University With sodium being the most abundant alkali metal on Earth, organosodium compounds are an attractive choice for sustainable chemical synthesis. However, organosodium compounds are rarely used-and are overshadowed by organolithium compounds-because of a lack of convenient and efficient preparation methods. Here we report a halogen-sodium exchange method to prepare a large variety of (hetero)aryl- and alkenylsodium compounds including tri- and tetrasodioarenes, many of them previously inaccessible by other methods. The key discovery is the use of a primary and bulky alkylsodium lacking beta-hydrogens, which retards undesired reactions, such as Wurtz-Fittig coupling and beta-hydrogen elimination, and enables efficient halogen-sodium exchange. The alkylsodium is readily prepared in situ from neopentyl chloride and an easy-to-handle sodium dispersion. We believe that the efficiency, generality, and convenience of the present method will contribute to the widespread use of organosodium in organic synthesis, ultimately contributing to the development of sustainable organic synthesis by rivalling the currently dominant organolithium reagents. Halogen-sodium exchange reactions with neopentyl sodium provides access to a range of aryl and alkenyl organosodium compounds in situ, as an alternative to organolithium reagents. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2021 Sodium Glucose Co-Transporter 2 Inhibitor Ameliorates Autophagic Flux Impairment on Renal Proximal Tubular Cells in Obesity Mice EN Kazuhiko Fukushima Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 1 2021 Marked Hypertriglyceridemia in a Patient with type 2 Diabetes Receiving SGLT2 Inhibitors 103 107 EN Mayumi Senoo Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Atsuhito Tone Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Yusuke Imai Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Satoko Watanabe Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Mitsuhiro Kaneto Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Yasuyuki Shimomura Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Sanae Teshigawara Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Tatsuaki Nakatou Department of Internal Medicine, Diabetes Center, Okayama Saiseikai General Hospital Case Report 10.18926/AMO/61441 A 43-year-old male with type 2 diabetes, under treatment with 5 mg/day of dapagliflozin, was referred to our hospital with upper left abdominal pain and marked hypertriglyceridemia (triglycerides [TGs], 5,960 mg/dl). He was also on a low-carbohydrate diet that promoted ketosis under sodium glucose cotransporter 2 (SGLT2) inhibitor administration. Polyacrylamide gel electrophoresis revealed a remarkable increase in very-low-den-sity lipoprotein, a TG-rich lipoprotein particle synthesized in the liver using free fatty acids derived from adi-pose tissue. Although SGLT2 inhibitors generally improve the lipid profile, under certain conditions such as a low-carbohydrate diet, they may adversely exacerbate the lipid profile via ketosis. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 1 2021 The Optimal Prepregnancy Body Mass Index for Lactation in Japanese Women with Neonatal Separation as Analyzed by a Differential Equation 63 69 EN Katsuhiko Tada Department of Obstetrics and Gynecology, National Hospital Organization, Okayama Medical Center Yasunari Miyagi Miyake Ofuku Clinic Kazue Nakamura Department of Neonatology, National Hospital Organization, Okayama Medical Center Moe Yorozu Department of Obstetrics and Gynecology, National Hospital Organization, Okayama Medical Center Emi Fukushima Department of Nursing, National Hospital Organization, Okayama Medical Center Kazumasa Kumazawa Department of Obstetrics and Gynecology, National Hospital Organization, Okayama Medical Center Makoto Nakamura Department of Neonatology, National Hospital Organization, Okayama Medical Center Misao Kageyama Department of Neonatology, National Hospital Organization, Okayama Medical Center Original Article 10.18926/ATM/61436 We used a differential equation to identify the biological relationship between the maternal prepregnancy body mass index (BMI) and lactation on postpartum day 4 in Japanese women with neonatal separation. This retro-spective observational study included 252 mothers (135 primiparas, 117 multiparas) whose singleton neonates were admitted to a neonatal ICU. We formulated hypotheses based on breast anatomy to analyze the relation-ship between the expressed milk obtained on postpartum day 4 and the maternal prepregnancy BMI with the following differential equation: y’(x) = k y(x)/x, where k is the constant, x is the prepregnancy BMI, and y is the expressed milk volume. The formula was then obtained as y(x) = axk, where a is the constant. The Akaike information criterion (AIC) was used to estimate the regression equation with the maximum likelihood for primiparas and multiparas. The best criteria for BMI determined by the AIC were 20.89 kg/m2 in primiparas and 20.19 kg/m2 in multiparas. These were the optimal BMI values for lactation, coinciding with the median prepregnancy BMI in the study population (20.78 kg/m2 in primiparas and 20.06 kg/m2 in multiparas). The formula based on biomathematics might help establish the biological relationship between prepregnancy BMI and breastmilk volume. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 75 1 2021 Possible Protective Effect of Remote Ischemic Preconditioning on Acute Kidney Injury Following Elective Percutaneous Coronary Intervention: Secondary Analysis of a Multicenter, Randomized Study 45 53 EN Hiroaki Otsuka Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Toru Miyoshi Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Kentaro Ejiri Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Kunihisa Kohno Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Makoto Nakahama Department of Cardiology, Fukuyama City Hospital Masayuki Doi Department of Cardiology, Kagawa Prefectural Central Hospital Mitsuru Munemasa Department of Cardiology, Okayama Medical Center Masaaki Murakami Department of Cardiology, Okayama Heart Clinic Kazufumi Nakamura Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Hiroshi Ito Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Original Article 10.18926/AMO/61433 Remote ischemic preconditioning (RIPC) is a promising strategy for protecting against ischemic reperfusion injury. This study is a secondary analysis of a randomized study that aimed to evaluate the effect of RIPC on the early increase in serum creatinine (SCr) following percutaneous coronary intervention (PCI), which is associ-ated with contrast-induced acute kidney injury. Patients with stable angina undergoing elective PCI were assigned to control, RIPC, and continuous infusion of nicorandil (nicorandil) groups. The endpoint of this study was the incidence of the early increase in SCr, a predictor of contrast-induced acute kidney injury, which was defined as either a > 20% or absolute increase by 0.3 mg/dl of SCr levels after 24 h of PCI. This study included 220 patients for whom a dataset of SCr values was available. The incidence of the early increase in SCr was significantly lower in the RIPC than in the control (1.3% vs 10.8%, p = 0.03) group, but was not significantly different between the nicorandil and control groups. In multivariate analysis, RIPC remained a significant fac-tor associated with a reduction in the incidence of early increase in SCr. RIPC reduces the incidence of early increase in SCr in patients with stable angina following elective PCI. No potential conflict of interest relevant to this article was reported.

Wiley Acta Medica Okayama 2047-9980 9 16 2020 Effect of Luseogliflozin on Heart Failure With Preserved Ejection Fraction in Patients With Diabetes Mellitus e015103 EN Kentaro Ejiri Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Toru Miyoshi Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Hajime Kihara Department of Internal Medicine, Kihara Cardiovascular Clinic Yoshiki Hata Department of Cardiology, Minamino Cardiovascular Hospital Toshihiko Nagano Department of Internal Medicine, Iwasa Hospital Atsushi Takaishi Department of Cardiology, Mitoyo General Hospital Hironobu Toda Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Seiji Nanba Department of Cardiology, Okayama Rosai Hospital Yoichi Nakamura Department of Cardiovascular Medicine, Specified Clinic of Soyokaze Cardiovascular Medicine and Diabetes Care Satoshi Akagi Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Satoru Sakuragi Department of Cardiovascular Medicine, Iwakuni Clinical Center Taro Minagawa Department of Internal Medicine, Minagawa Cardiovascular Clinic Yusuke Kawai Department of Cardiovascular Medicine, Okayama City Hospital Nobuhiro Nishii Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Soichiro Fuke Department of Cardiovascular Medicine, Japanese Red Cross Okayama Hospital Masaki Yoshikawa Department of Cardiology, Fukuyama City Hospital Kazufumi Nakamura Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Hiroshi Ito Okayama University Graduate School of Medicine, Density and Pharmaceutical Sciences Background Effects of sodium‐glucose cotransporter 2 inhibitors on reducing hospitalization for heart failure have been reported in randomized controlled trials, but their effects on patients with heart failure with preserved ejection fraction (HFpEF) are unknown. This study aimed to evaluate the drug efficacy of luseogliflozin, a sodium‐glucose cotransporter 2 inhibitor, in patients with type 2 diabetes mellitus and HFpEF. Methods and Results We performed a multicenter, open‐label, randomized, controlled trial for comparing luseogliflozin 2.5 mg once daily with voglibose 0.2 mg 3 times daily in patients with type 2 diabetes mellitus suffering from HFpEF (left ventricular ejection fraction >45% and BNP [B‐type natriuretic peptide] concentrations ≥35 pg/mL) in a 1:1 randomization fashion. The primary outcome was the difference from baseline in BNP levels after 12 weeks of treatment between the 2 drugs. A total of 173 patients with diabetes mellitus and HFpEF were included. Of these, 83 patients were assigned to receive luseogliflozin and 82 to receive voglibose. There was no significant difference in the reduction in BNP concentrations after 12 weeks from baseline between the 2 groups. The ratio of the mean BNP value at week 12 to the baseline value was 0.79 in the luseogliflozin group and 0.87 in the voglibose group (percent change, －9.0% versus －1.9%; ratio of change with luseogliflozin versus voglibose, 0.93; 95% CI, 0.78–1.10; P=0.26). Conclusion In patients with type 2 diabetes mellitus and HFpEF, there is no significant difference in the degree of reduction in BNP concentrations after 12 weeks between luseogliflozin and voglibose. No potential conflict of interest relevant to this article was reported.

Royal Society of Chemistry Acta Medica Okayama 2040-3364 12 42 2020 Iron nanoparticle templates for constructing 3D graphene framework with enhanced performance in sodium-ion batteries 21780 21787 EN Benoît D. L. Campéon Graduate School of Natural Science and Technology, Okayama University Chen Wang Research Core for Interdisciplinary Sciences, Okayama University Yuta Nishina Graduate School of Natural Science and Technology, Okayama University This study examines the synthesis and electrochemical performance of three-dimensional graphene for Li-ion batteries and Na-ion batteries. The in situ formation of iron hydroxide nanoparticles (Fe(OH)x NPs) of various weights on the surface of graphene oxide, followed by thermal treatment at elevated temperature and washing using hydrochloric acid, furnished 3D graphene. The characterization studies confirmed the prevention of graphene layer stacking by over 90% compared with thermal treatment without Fe(OH)x. The electrochemical performance of the 3D graphene was evaluated as a counter electrode for lithium metal and sodium metal in a half-cell configuration. This material showed good performances with a charging capacity of 507 mA h g－1 at 372 mA g－1 in Li-ion batteries and 252 mA h g－1 at 100 mA g－1 in Na-ion batteries, which is 1.4 and 1.9 times higher, respectively, than the graphene prepared without Fe(OH)x templates. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2020 The relationship between repeated measurement of casual and 24-h urinary sodium-to-potassium ratio in patients with chronic kidney disease EN Yuka Okuyama Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 2073-4409 9 10 2020 In Vitro Studies to Define the Cell-Surface and Intracellular Targets of Polyarginine-Conjugated Sodium Borocaptate as a Potential Delivery Agent for Boron Neutron Capture Therapy 2149 EN Atsushi Fujimura Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Seiji Yasui Neutron Therapy Research Center, Okayama University Kazuyo Igawa Neutron Therapy Research Center, Okayama University Ai Ueda Neutron Therapy Research Center, Okayama University Kaori Watanabe Neutron Therapy Research Center, Okayama University Tadashi Hanafusa Neutron Therapy Research Center, Okayama University Yasuaki Ichikawa Neutron Therapy Research Center, Okayama University Sachiko Yoshihashi Graduate School of Engineering, Nagoya University Kazuki Tsuchida Graduate School of Engineering, Nagoya University Atsunori Kamiya Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shuichi Furuya Neutron Therapy Research Center, Okayama University Boron neutron capture therapy (BNCT) requires pharmaceutical innovations and molecular-based evidence of effectiveness to become a standard cancer therapeutic in the future. Recently, in Japan, 4-borono-L-phenylalanine (BPA) was approved as a boron agent for BNCT against head and neck (H&N) cancers. H&N cancer appears to be a suitable target for BPA-BNCT, because the expression levels of L-type amino acid transporter 1 (LAT1), one of the amino acid transporters responsible for BPA uptake, are elevated in most cases of H&N cancer. However, in other types of cancer including malignant brain tumors, LAT1 is not always highly expressed. To expand the possibility of BNCT for these cases, we previously developed poly-arginine peptide (polyR)-conjugated mercaptoundecahydrododecaborate (BSH). PolyR confers the cell membrane permeability and tumor selectivity of BSH. However, the molecular determinants for the properties are not fully understood. In this present study, we have identified the cluster of differentiation 44 (CD44) protein and translational machinery proteins as a major cell surface target and intracellular targets of BSH-polyR, respectively. CD44, also known as a stem cell-associated maker in various types of cancer, is required for the cellular uptake of polyR-conjugated molecules. We showed that BSH-polyR was predominantly delivered to a CD44(High) cell population of cancer cells. Once delivered, BSH-polyR interacted with the translational machinery components, including the initiation factors, termination factors, and poly(A)-biding protein (PABP). As a proof of principle, we performed BSH-polyR-based BNCT against glioma stem-like cells and revealed that BSH-polyR successfully induced BNCT-dependent cell death specifically in CD44(High) cells. Bioinformatics analysis indicated that BSH-polyR would be suitable for certain types of malignant tumors. Our results shed light on the biochemical properties of BSH-polyR, which may further contribute to the therapeutic optimization of BSH-BNCT in the future. No potential conflict of interest relevant to this article was reported.

BMFH Press Acta Medica Okayama 2186-3342 39 3 2020 Cyclic nigerosylnigerose ameliorates DSS-induced colitis with restoration of goblet cell number and increase in IgA reactivity against gut microbiota in mice 188 196 EN Takeshi Tsuruta Laboratory of Animal Nutrition, Graduate School of Environmental and Life Science, Okayama University Emiko Katsumata Laboratory of Animal Nutrition, Graduate School of Environmental and Life Science, Okayama University Akiko Mizote Hayashibara Co., Ltd. Hou Jian Jian Laboratory of Animal Nutrition, Graduate School of Environmental and Life Science, Okayama University Teresia Aluoch Muhomah Laboratory of Animal Nutrition, Graduate School of Environmental and Life Science, Okayama University Naoki Nishino Laboratory of Animal Nutrition, Graduate School of Environmental and Life Science, Okayama University Cyclic nigerosylnigerose (CNN) is a cyclic oligosaccharide. Oral administration of CNN promotes immunoglobulin A (IgA) secretion in the gut. IgA is a major antibody secreted into the gut and plays a crucial role in suppressing gut inflammation due to commensal gut microbiota. To investigate the effect of administration of CNN to promote IgA secretion on gut inflammation, experimental colitis was induced with dextran sulfate sodium (DSS) in Balb/c mice after 6 weeks of CNN pre-feeding. The severity of colitis was evaluated based on a disease activity index (DAI), the gene expression of inflammatory cytokines, and a histological examination. The CNN-treated mice with DSS-induced colitis (CNN-DSS group) showed significantly lower DAI scores and mRNA levels of interleukin-1 compared with the CNN-untreated mice with DSS-induced colitis (DSS group). Histological examination of the colon revealed that the pathological score was significantly lower in the CNN-DSS group compared with the DSS group due to the reduced infiltration of immune cells. The number of goblet cells was significantly higher in the CNN-DSS group compared with the DSS group. The IgA concentration and the ratio of microbiota coated with IgA were evaluated in the cecal content. Although there was no difference in the IgA concentration among groups, a higher proportion of cecal microbiota were coated with IgA in the CNN-DSS group compared with that in the DSS group. These results suggest that CNN might preserve goblet cells in the colon and promote IgA coating of gut microbiota, which synergistically ameliorate gut inflammation in mice with DSS-induced colitis. 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.

Okayama University Medical School Acta Medica Okayama 0386-300X 74 4 2020 Relationship Between Partial Carbon Dioxide Pressure and Strong Ions in Humans: A Retrospective Study 319 325 EN Satoshi Isoyama Department of Anesthesiology and Resuscitation, Okayama University Hospital Satoshi Kimura Department of Anesthesiology and Resuscitation, Okayama University Hospital Hiroshi Morimatsu Department of Anesthesiology and Resuscitation, Okayama University Hospital Original Article 10.18926/AMO/60370 Little is known about the role of a strong ions in humans with respiratory abnormalities. In this study, we investigated the associations between partial carbon dioxide pressure (pCO2) and each of sodium ion (Na+) concentrations, chloride ion (Cl－) concentrations and their difference (SIDNa-Cl). Blood gas data were obtained from patients in a teaching hospital intensive care unit between August 2013 and January 2017. The association between pCO2 and SIDNa-Cl was defined as the primary outcome. The associations between pCO2 and [Cl－], [Na+] and other strong ions were secondary outcomes. pCO2 was stratified into 10 mmHg-wide bands and treated as a categorical variable for comparison. As a result, we reviewed 115,936 blood gas data points from 3,840 different ICU stays. There were significant differences in SIDNa-Cl, [Cl－], and [Na+] among all categorized pCO2 bands. The respective pCO2 SIDNa-Cl, [Cl－], and [Na+] correlation coefficients were 0.48, －0.31, and 0.08. SIDNa-Cl increased and [Cl－] decreased with pCO2, with little relationship between pCO2 and [Na+] across subsets. In conclusion, we found relatively strong correlations between pCO2 and SIDNa-Cl in the multiple blood gas datasets examined. Correlations between pCO2 and chloride concentrations, but not sodium concentrations, were further found to be moderate in these ICU data. No potential conflict of interest relevant to this article was reported.

Royal Society of Chemistry Acta Medica Okayama 2050-7488 8 29 2020 Mechanisms for overcharging of carbon electrodes in lithium-ion/sodium-ion batteries analysed by operando solid-state NMR 14472 14481 EN Kazuma Gotoh Graduate School of Natural Science & Technology, Okayama University Tomu Yamakami Graduate School of Natural Science & Technology, Okayama University Ishin Nishimura Graduate School of Natural Science & Technology, Okayama University Hina Kometani Graduate School of Natural Science & Technology, Okayama University Hideka Ando Graduate School of Natural Science & Technology, Okayama University Kenjiro Hashi National Institute for Materials Science Tadashi Shimizu National Institute for Materials Science Hiroyuki Ishida Graduate School of Natural Science & Technology, Okayama University A precise understanding of the mechanism for metal (Li and Na) plating on negative electrodes that occurs with overcharging is critical to managing the safety of lithium- and sodium-ion batteries. In this work, an in-depth investigation of the overlithiation/oversodiation and subsequent delithiation/desodiation of graphite and hard carbon electrodes in the first cycle was conducted using operando7Li/23Na solid-state NMR. In the 7Li NMR spectra of half cells of carbon electrodes and metal counter electrodes, three types of signals corresponding to Li dendrites that formed on the surface of graphite, hard carbon, and the counter electrode were distinguished from the signal of Li metal foil of the counter electrode by applying an appropriate orientation of the testing cell. For graphite overlithiation, the deposition of Li dendrites started immediately or soon after the minimum electric potential in the lithiation curve. In contrast, the deposition of Li dendrites in hard carbon started after the end of quasimetallic lithium formation for overlithiation at rates below 3.0C. Similar behaviour was also observed for the oversodiation of hard carbon. The formation of quasimetallic Li or Na in the pores of hard carbon serves as a buffer for the metal plating that occurs with overcharging of the batteries. Furthermore, some of the deposited Li/Na dendrites contribute to reversible capacities. A mechanism for the inhomogeneous disappearance of quasimetallic Li during delithiation of hard carbon is also proposed. No potential conflict of interest relevant to this article was reported.

American Chemical Society Acta Medica Okayama 0743-7463 30 9 2014 Microfluidic Approach to the Formation of Internally Porous Polymer Particles by Solvent Extraction 2470 2479 EN Takaichi Watanabe Department of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University Carlos G. Lopez Department of Chemical Engineering, Imperial College London Jack F. Douglas Materials Science and Engineering Division, National Institute of Standards and Technology Tsutomu Ono Department of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University João T. Cabral Department of Chemical Engineering, Imperial College London We report the controlled formation of internally porous polyelectrolyte particles with diameters ranging from tens to hundreds of micrometers through selective solvent extraction using microfluidics. Solvent-resistant microdevices, fabricated by frontal photopolymerization, encapsulate binary polymer (P)/solvent (S1) mixtures by a carrier solvent phase (C) to form plugs with well-defined radii and low polydispersity; the suspension is then brought into contact with a selective extraction solvent (S2) that is miscible with C and S1 but not P, leading to the extraction of S1 from the droplets. The ensuing phase inversion yields polymer capsules with a smooth surface but highly porous internal structure. Depending on the liquid extraction time scale, this stage can be carried out in situ, within the chip, or ex situ, in an external S2 bath. Bimodal polymer plugs are achieved using asymmetrically inverted T junctions. For this demonstration, we form sodium poly(styrenesulfonate) (P) particles using water (S1), hexadecane (C), and methyl ethyl ketone (S2). We measure droplet extraction rates as a function of drop size and polymer concentration and propose a simple scaling model to guide particle formation. We find that the extraction time required to form particles from liquid droplets does not depend on the initial polymer concentration but is rather proportional to the initial droplet size. The resulting particle size follows a linear relationship with the initial droplet size for all polymer concentrations, allowing for the precise control of particle size. The internal particle porous structure exhibits a polymer density gradient ranging from a dense surface skin toward an essentially hollow core. Average particle porosities between 10 and 50% are achieved by varying the initial droplet compositions up to 15 wt % polymer. Such particles have potential applications in functional, optical, and coating materials. No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 21 11 2020 Sodium Glucose Co-Transporter 2 Inhibitor Ameliorates Autophagic Flux Impairment on Renal Proximal Tubular Cells in Obesity Mice 4054 EN Kazuhiko Fukushima Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shinji Kitamura Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kenji Tsuji Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yizhen Sang Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Jun Wada Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Obesity is supposed to cause renal injury via autophagy deficiency. Recently, sodium glucose co-transporter 2 inhibitors (SGLT2i) were reported to protect renal injury. However, the mechanisms of SGLT2i for renal protection are unclear. Here, we investigated the effect of SGLT2i for autophagy in renal proximal tubular cells (PTCs) on obesity mice. We fed C57BL/6J mice with a normal diet (ND) or high-fat and -sugar diet (HFSD) for nine weeks, then administered SGLT2i, empagliflozin, or control compound for one week. Each group contained N = 5. The urinary N-acetyl-beta-d-glucosaminidase level in the HFSD group significantly increased compared to ND group. The tubular damage was suppressed in the SGLT2i-HFSD group. In electron microscopic analysis, multi lamellar bodies that increased in autophagy deficiency were increased in PTCs in the HFSD group but significantly suppressed in the SGLT2i group. The autophagosomes of damaged mitochondria in PTCs in the HFSD group frequently appeared in the SGLT2i group. p62 accumulations in PTCs were significantly increased in HFSD group but significantly suppressed by SGLT2i. In addition, the mammalian target of rapamycin was activated in the HFSD group but significantly suppressed in SGLT2i group. These data suggest that SGLT2i has renal protective effects against obesity via improving autophagy flux impairment in PTCs on a HFSD. No potential conflict of interest relevant to this article was reported.

Springer Acta Medica Okayama 2363-9024 5 1 2019 Anesthetic management of a patient with sodium-channel myotonia: a case report 77 EN Naohisa Matsumoto Department of Anesthesiology and Resuscitology, Okayama University Hospital Rei Nishimoto Department of Anesthesiology and Resuscitology, Okayama University Hospital Yoshikazu Matsuoka Departments of Anesthesiology and Resuscitology, Okayama University Graduate School of Medicine Yoshimasa Takeda Department of Anesthesiology and Resuscitology, Okayama University Hospital Hiroshi Morimatsu Department of Anesthesiology and Resuscitology, Okayama University Hospital Background: Sodium-channel myotonia (SCM) is a nondystrophic myotonia, characterized by pure myotonia without muscle weakness or paramyotonia. The prevalence of skeletal muscle channelopathies is approximately 1 in 100,000, and the prevalence of SCM is much lower. To our knowledge, this is the first report on anesthetic management of a patient with SCM. Case presentation: A 23-year-old woman with congenital nasal dysplasia and SCM was scheduled to undergo rhinoplasty with autologous costal cartilage. Total intravenous anesthesia without muscle relaxants was administered followed by continuous intercostal nerve block. Although transient elevation of potassium level in the blood was observed during surgery, the patient did not show exacerbation of myotonic or paralytic symptoms in the postoperative period. Conclusion: Total intravenous anesthesia and peripheral nerve block can be administered safely to a patient with SCM. However, careful monitoring of the symptoms and electrolytes is recommended. No potential conflict of interest relevant to this article was reported.

American Institute of Physics Acta Medica Okayama 2158-3226 9 12 2019 Magnetic characterization change by solvents of magnetic nanoparticles in liquid-phase magnetic immunoassay 125317 EN Katsuya Jinno Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Bunta Hiramatsu Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Kenta Tsunashima Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Kayo Fujimoto Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Kenji Sakai Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Toshihiko Kiwa Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Keiji Tsukada Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Liquid-phase magnetic immunoassay (MIA) using magnetic nano-particles (MNPs) has been studied as a more rapid method compared to optical methods for inspecting proteins and viruses. MIA can estimate the number of conjugated antibodies without being washed differently from conventional optical immunoassay. However, in the case of the liquid phase, it is considered that the magnetic properties of MNPs are affected by physical properties such as viscosity and impurity substances such as biological substances contained in the blood. In this study, the effect of sodium chloride (NaCl) in buffer and serum solution was evaluated to reveal the effect of serum because the sodium (Na+) and chloride (Cl-) ions in the serum dominate ion balance of blood. The measurement results of AC magnetic susceptibility and a dynamic light scattering (DLS) showed that the aggregation of MNPs was largely affected by the concentration of NaCl. This effect of the NaCl could be explained by shielding of the surface charge of MNPs by ions in the solution. Although the concentrations of NaCl in the buffer and serum solution were almost same, we found that MNPs were aggregated more in their size for those in the serum solution because of other impurities, such as proteins. These results suggest evaluation of effects of the contaminants in serum and optimization of polymer coatings of MNPs could be important factors to realize measurements of magnetic immunoassay with high accuracy. (C) 2019 Author(s). No potential conflict of interest relevant to this article was reported.

Springer Acta Medica Okayama 2199-3823 5 4 2019 Separation Between Silicon and Aluminum Powders Contained Within Pulverized Scraped Silicon-Based Waste Solar Cells by Flotation Method 551 560 EN Sho Harada Department of Material and Energy Science, Graduate School of Environmental and Life Science, Okayama University Md. Azhar Uddin Department of Material and Energy Science, Graduate School of Environmental and Life Science, Okayama University Yoshiei Kato Department of Material and Energy Science, Graduate School of Environmental and Life Science, Okayama University Takanori Kawanishi Wet Process Division, Toho Kasei Co., Ltd Yoshiaki Hayashi Wet Process Division, Toho Kasei Co., Ltd There are few study examples on the separation of metals by floating method. In this study, separation of silicon and aluminum, which are the main components of silicon-based solar cell module, was carried out by floating method in order to purify silicon from waste solar cell module. The selection of surfactant, control of electric charge, wettability of the solid particles, surface tensions, and bubble surface area are important for separation of solids by floating method. Sodium dodecyl sulfate (SDS) can increase the hydrophobicity of aluminum powder due to the difference of surface potentials between silicon and aluminum. SDS behaves as a collector of aluminum as well as a frothing agent to decrease the bubble size. At a SDS concentration of 2 g/L and sample dipping time of 10 min, 80.1 mass% of aluminum was floated and separated, and the sedimentary silicon reached a purity of 90.7% from a mixture of 50 mass% aluminum and 50 mass% silicon. Finally, at a pH value of 7.0, SDS concentration between 1.0 and 2.5 g/L and air flow rate of 2.5 L/min (STP) were suitable experimental conditions to purify silicon from a mixture of silicon and aluminum by flotation separation method. No potential conflict of interest relevant to this article was reported.

TAYLOR & FRANCIS Acta Medica Okayama 0916-8451 83 5 2019 Mulberry juice freeze-dried powder attenuates the disease severity by the maintaining of colon mucosa in mice with DSS-induced acute colitis 914 922 EN Yang Wang Graduate School of Environmental and Life Science , Okayama University Toshimitsu Hatabu Graduate School of Environmental and Life Science , Okayama University This study aimed to evaluate the microbial compositions and gene expression related to inflammation in dextran sodium sulfate (DSS)-induced acute colitis and the effect of mulberry supplementation. Male BALB/c mice received a diet supplemented with mulberry juice freeze-dried powder (MFP) or not for 3 weeks. After 3 weeks, the mice received water containing 5% (w/v) DSS or not for 1 week. The disease activity index score in mice fed MFP was significantly decreased. A significant decrease in Bifidobacterium spp. and the Clostridium perfringens subgroup was observed in mice not fed MFP. The number of goblet cell and NLRP6 expression were observed in mice fed a diet supplemented with MFP compared with mice not fed MFP. These results may indicate that mulberry mitigates DSS-induced acute colitis by a changing the gut microbial flora and by improving mucosal conditions. No potential conflict of interest relevant to this article was reported.

Blackwell Acta Medica Okayama 0013-9580 49 3 2007 Rasmussen encephalitis associated with SCN1A mutation 521 526 EN Iori Ohmori Departments of Cellular Physiology, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Mamoru Ouchida Departments of Molecular Genetics, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Katsuhiro Kobayashi Departments of Child Neurology, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Yoshimi Jitsumori Takushi Inoue Departments of Child Neurology, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Kenji Shimizu Departments of Molecular Genetics, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Hideki Matsui Departments of Cellular Physiology, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Yoko Ohtsuka Departments of Child Neurology, Graduate School of Medicine,Dentistry and Pharmaceutical Sciences, Okayama University Yoshihiro Maegaki Division of Child Neurology, Institute of Neurological Sciences, Faculty of Medicine, Tottori University Mutations in the SCN 1 A gene, encoding the neuronal voltage-gated sodium channel alpha1 subunit, cause SMEI, GEFS+, and related epileptic syndromes. We herein report the R1575C-SCN 1 A mutation identified in a patient with Rasmussen encephalitis. R1575C were constructed in a recombinant human SCN 1 A and then heterologously expressed in HEK293 cells along with the human beta1 and beta2 sodium channel accessory subunits. Whole-cell patch-clamp recording was used to define biophysical properties. The R1575C channels exhibited increased channel availability and an increased persistent sodium current in comparison to the wild-type. These defects of electrophysiological properties can result in neuronal hyperexitability. The seizure susceptibility allele may influence the pathogenesis of Rasmussen encephalitis in this case. No potential conflict of interest relevant to this article was reported.

American Chemical Society Acta Medica Okayama 24701343 4 12 2019 Characterization of Pieces of Paper That Form Reagent Containers for Use as Portable Analytical Devices 15249 15254 EN Supatana Buking Flow Innovation-Research for Science and Technology Laboratories (FIRST Labs) and Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Mahidol University Yusuke Suedomi Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University Duangjai Nacapricha Flow Innovation-Research for Science and Technology Laboratories (FIRST Labs) and Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Mahidol University Takashi Kaneta Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University Reagent-deposited pieces of paper were characterized by the use of a compact conductometer, a compact pH sensor, and a conventional spectrophotometer to assess their suitability for use as reagent containers. The pieces of paper were fabricated by wax printing to form a limited hydrophilic area to which a consistent volume of an aqueous reagent could be added. The pieces of paper without the reagent increased the conductivity of water gradually because of the release of sodium salts, whereas pH of NaOH decreased because of the acidity of the functional groups in the paper. Three reagents, sulfamic acid as an acid, Na2CO3 as a base, and BaCl2 as a metal salt, were deposited on the pieces of paper to evaluate their ability to release from the pieces of paper. Sulfamic acid and Na2CO3 were released in quantities of 58 and 73% into water after 420 s, whereas 100% of BaCl2 was released after 480 s. The conductometric titrations of NaOH, HCl, and Na2SO4, and the spectrophotometry of Fe2+ were examined using the pieces of paper that contained sulfamic acid, Na2CO3, BaCl2, and 1,10-phenanthroline. Titrations using the pieces of paper suggested that the reagents were quantitatively released into the titrant, which resulted in a linear relationship between the endpoints and the equivalent points. In 120 s of soaking time, 60-70% of the reagents were released. The spectrophotometric measurements of Fe2+ indicated that when an excess amount of the reagents was deposited onto the pieces of paper, they nonetheless sufficiently fulfilled the role of a reagent container. No potential conflict of interest relevant to this article was reported.

Academic Press Acta Medica Okayama 09699961 50 2013 CACNA1A variants may modify the epileptic phenotype of Dravet syndrome 209 217 EN Iori Ohmori Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Mamoru Ouchida Department of Molecular Genetics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Katsuhiro Kobayashi Yoshimi Jitsumori Akiko Mori Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Hiroyuki Michiue Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Teiichi Nishiki Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Yoko Ohtsuka Hideki Matsui Department of Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University Dravet syndrome is an intractable epileptic syndrome beginning in the first year of life. De novo mutations of SCN1A, which encode the Na(v)1.1 neuronal voltage-gated sodium channel, are considered the major cause of Dravet syndrome. In this study, we investigated genetic modifiers of this syndrome. We performed a mutational analysis of all coding exons of CACNA1A in 48 subjects with Dravet syndrome. To assess the effects of CACNA1A variants on the epileptic phenotypes of Dravet syndrome, we compared clinical features in two genotype groups: 1) subjects harboring SCN1A mutations but no CACNA1A variants (n=20) and 2) subjects with SCN1A mutations plus CACNA1A variants (n=20). CACNA1A variants detected in patients were studied using heterologous expression of recombinant human Ca(v)2.1 in HEK 293 cells and whole-cell patch-clamp recording. Nine CACNA1A variants, including six novel ones, were detected in 21 of the 48 subjects (43.8%). Based on the incidence of variants in healthy controls, most of the variants seemed to be common polymorphisms. However, the subjects harboring SCN1A mutations and CACNA1A variants had absence seizures more frequently than the patients with only SCN1A mutations (8/20 vs. 0/20, p=0.002). Moreover, the former group of subjects exhibited earlier onset of seizures and more frequent prolonged seizures before one year of age, compared to the latter group of subjects. The electrophysiological properties of four of the five novel Ca(v)2.1 variants exhibited biophysical changes consistent with gain-of-function. We conclude that CACNA1A variants in some persons with Dravet syndrome may modify the epileptic phenotypes. 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 72 6 2018 Changes in Serum Biochemical Markers in Relation to Chief Complaints and Aging in General Medicine 553 562 EN Daisuke Omura Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Asuka Sato Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kosuke Oka Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yoshihisa Hanayama Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroko Ogawa Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mikako Obika 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/56372 To clarify potential relationships between chief complaints of patients and laboratory data with a focus on aging-related changes, we retrospectively analyzed the data of 843 patients who visited a general medicine department for the first time. Their chief complaints were classified into 8 major symptoms: visceral pain, somatic pain, fever, cough, dizziness, fatigue, appetite loss, and edema. We compared the laboratory data obtained from the patients with complaints with the data of symptom-free (control) patients. The serum sodium and potassium levels in the fever group were decreased compared to those in the control group. In the fever group, the serum sodium level was inversely correlated with age. The ratio of serum urea nitrogen to creatinine (UN/Cr) was increased in the appetite-loss group. There were significant age-dependent increases in the UN/Cr ratio in the appetite-loss and edema groups. Of note, serum levels of free thyroxin were lower in the dizziness group compared to the control group. In addition, the free thyroxin level was inversely correlated with age in the dizziness group but not in the asymptomatic control group. Collectively, the results indicated that osmolality-related laboratory data are strongly associated with individual primary symptoms at the first visit regardless of the final diagnosis. The consideration of age-dependent changes of these markers is helpful for diagnosing latent disorders based on various primary symptoms. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2018 Prognostic Significance of the Sodium Channel Blocker Test in Patients With Brugada Syndrome EN Akira Ueoka Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 72 5 2018 Clinical Characteristics of Febrile Outpatients : Possible Involvement of Thyroid Dysfunction in Febrile Tachycardia 447 456 EN Kosuke Oka Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yoshihisa Hanayama Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Asuka Sato Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Daisuke Omura Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Miho Yasuda Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ko Hasegawa Department of General Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Mikako Obika 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/56242 We retrospectively analyzed the cases of 148 febrile patients whose body temperature (BT) was ≥ 37.5°C at our hospital. We categorized them into seven groups; those with bacterial and viral infection, nonspecific inflammation, neoplasm, connective tissue disease (CTD), drug-induced disease, and unidentified causes. Our analysis revealed that the patient’s BT at the 1st visit (BT-1st visit) and highest BT during the febrile period (BT-max) differed significantly among all categories except neoplasm. The greatest difference between BT-1st visit and BT-max was highest in the CTD group (1.5°C). Positive correlations of heart rate and C-reactive protein (CRP) level with BT-max and a negative correlation between serum sodium level with BT-max were uncovered. The serum thyroid-stimulating hormone (TSH) level and the ratio of TSH/free thyroxine were negatively correlated with BT-max, especially in the viral infection group, suggesting the existence of occult thyrotoxicosis in accord with a febrile condition, possibly leading to febrile tachycardia. A febrile gap between BT-1st visit and BT-max (except in the neoplasm group) was shown, in which BT-max was correlated with thyroid function. Clinicians should recognize the fluctuation of BT when diagnosing febrile patients, and tachycardia in such febrile patients may be, at least in part, associated with subclinical thyroid dysfunction. No potential conflict of interest relevant to this article was reported.

Dove Medical Press Acta Medica Okayama 1176-6344 11 2015 Epoprostenol sodium for treatment of pulmonary arterial hypertension 265 270 EN Yukihiro Saito Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kazufumi Nakamura Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Satoshi Akagi Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshihiro Sarashina Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Kentaro Ejiri Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Aya Miura Division of Cardiology, National Hospital Organization Okayama Medical Center Aiko Ogawa Division of Cardiology, National Hospital Organization Okayama Medical Center Hiromi Matsubara Division of Cardiology, National Hospital Organization Okayama Medical Center Hiroshi Ito Department of Cardiovascular Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences 　The release of endogenous prostacyclin (PGI2) is depressed in patients with pulmonary arterial hypertension (PAH). PGI2 replacement therapy by epoprostenol infusion is one of the best treatments available for PAH. Here, we provide an overview of the current clinical data for epoprostenol. Epoprostenol treatment improves symptoms, exercise capacity, and hemodynamics, and is the only treatment that has been shown to reduce mortality in patients with idiopathic PAH (IPAH) in randomized clinical trials. We have reported that high-dose epoprostenol therapy (>40 ng/kg/min) also results in marked hemodynamic improvement in some patients with IPAH. High-dose epoprostenol has a pro-apoptotic effect on PAH-PASMCs via the IP receptor and upregulation of Fas ligand (FasL) in vitro. However, long-term intravenous administration of epoprostenol is sometimes associated with catheter-related infections and leads to considerable inconvenience for the patient. In the future, the development of new routes of administration or the development of powerful PGI2 analogs, IP-receptor agonists, and gene and cell-based therapy enhancing PGI2 production with new routes of administration is required. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 129 1 2017 水素水は怪しい水でしょうか？ 9 15 EN Atsunori Nakao Department of Emergency and Critical Care Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 71 1 2017 Syndrome of Inappropriate Antidiuretic Hormone Secretion Following Liver Transplantation 85 89 EN Kosei Takagi Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takahito Yagi Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Susumu Shinoura Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Yuzo Umeda Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ryuichi Yoshida Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Daisuke Nobuoka Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Nobuyuki Watanabe Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Takashi Kuise Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Tomokazu Fuji Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Hiroyuki Araki Department of Gastroenterological 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 Case Report 10.18926/AMO/54830 Syndrome of inappropriate antidiuretic hormone secretion (SIADH) is an extremely rare cause of hyponatremia post-liver transplantation. A 15-year-old Japanese girl with recurrent cholangitis after Kasai surgery for biliary atresia underwent successful living donor liver transplantation. Peritonitis due to gastrointestinal perforation occurred. Hyponatremia gradually developed but improved after hypertonic sodium treatment. One month later, severe hyponatremia rapidly recurred. We considered the hyponatremia’s cause as SIADH. We suspected that tacrolimus was the disease’s cause, so we used cyclosporine instead, plus hypertonic sodium plus water intake restriction, which improved the hyponatremia. Symptomatic hyponatremia manifested by SIADH is a rare, serious complication post-liver transplantation. No potential conflict of interest relevant to this article was reported.

ROYAL SOC CHEMISTRY Acta Medica Okayama 2050-7488 4 34 2016 Combination of solid state NMR and DFT calculation to elucidate the state of sodium in hard carbon electrodes 13183 13193 EN Ryohei Morita Graduate School of Natural Science & Technology, Okayama University Kazuma Gotoh Graduate School of Natural Science & Technology, Okayama University Mika Fukunishi Elements Strategy Initiative for Catalysts and Batteries (ESICB), Kyoto University Kei Kubota Elements Strategy Initiative for Catalysts and Batteries (ESICB), Kyoto University Shinichi Komaba Elements Strategy Initiative for Catalysts and Batteries (ESICB), Kyoto University Naoto Nishimura Department of Chemistry and Materials Technology, Kyoto Institute of Technology Takashi Yumura Department of Chemistry and Materials Technology, Kyoto Institute of Technology Kenzo Deguchi National Institute for Materials Science Shinobu Ohki National Institute for Materials Science Tadashi Shimizue National Institute for Materials Science Hiroyuki Ishida Graduate School of Natural Science & Technology, Okayama University We examined the state of sodium electrochemically inserted in HC prepared at 700–2000 °C using solid state Na magic angle spinning (MAS) NMR and multiple quantum (MQ) MAS NMR. The 23Na MAS NMR spectra of Na-inserted HC samples showed signals only in the range between +30 and －60 ppm. Each observed spectrum was ascribed to combinations of Na+ ions from the electrolyte, reversible ionic Na components, irreversible Na components assigned to solid electrolyte interphase (SEI) or non-extractable sodium ions in HC, and decomposed Na compounds such as Na2CO3. No quasi-metallic sodium component was observed to be dissimilar to the case of Li inserted in HC. MQMAS NMR implies that heat treatment of HC higher than 1600 °C decreases defect sites in the carbon structure. To elucidate the difference in cluster formation between Na and Li in HC, the condensation mechanism and stability of Na and Li atoms on a carbon layer were also studied using DFT calculation. Na3 triangle clusters standing perpendicular to the carbon surface were obtained as a stable structure of Na, whereas Li2 linear and Li4 square clusters, all with Li atoms being attached directly to the surface, were estimated by optimization. Models of Na and Li storage in HC, based on the calculated cluster structures were proposed, which elucidate why the adequate heat treatment temperature of HC for high-capacity sodium storage is higher than the temperature for lithium storage. No potential conflict of interest relevant to this article was reported.

Elsevier Science B.V. Acta Medica Okayama 0021-9673 1440 2016 Direct determination of lignin peroxidase released from Phanerochaete chrysosporium by in-capillary enzyme assay using micellar electrokinetic chromatography 145 149 EN Airi Harada Keiko Sasaki Takashi Kaneta Here we describe the application of an in-capillary enzyme assay using micellar electrokinetic chromatography (MEKC) in the determination of enzyme activity in a crude culture medium containing lignin peroxidase released from Phanerochaete chrysosporium (P. chrysosporium). The method consists of a plug–plug reaction between lignin peroxidase and its substrate, veratryl alcohol, the separation of the product, veratraldehyde, from the other components including the enzyme and the culture medium, and the determination of the enzyme activity from the peak area of veratraldehyde produced by the plug–plug reaction. This method is more sensitive than conventional spectrophotometry since the background originates from the enzyme and the culture medium can be removed via MEKC separation. Veratraldehyde was separated at －10 kV in a background electrolyte containing 50 mM tartrate buffer (pH 2.5) and 50 mM sodium dodecyl sulfate (SDS) after a plug–plug reaction in the capillary for 5 min. The calibration curve of veratraldehyde was linear up to 4 pmol (500 μM) with a limit to quantification of 0.026 pmol (3.2 μM) (SN = 10). The activity of lignin peroxidase was directly measured from the peak area of veratraldehyde. The activity of lignin peroxidase released from P. chrysosporium into the medium for 7 days was successfully determined to be 3.40 UL－1. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 69 4 2015 Prevalence, Risk Factors, and Short-term Consequences of Traumatic Brain Injury-associated Hyponatremia 213 218 EN Tetsuya Yumoto Keiji Sato Toyomu Ugawa Shingo Ichiba Yoshihito Ujike Original Article 10.18926/AMO/53557 Hyponatremia, a common electrolyte disorder associated with traumatic brain injuries (TBIs), has high mortality and morbidity rates. The aim of this study was to identify the risk factors for hyponatremia associated with TBI. We retrospectively analyzed the cases of TBI patients who were admitted to the emergency intensive care unit at Okayama University Hospital between October 2011 and September 2014. A total of 82 TBI patients were enrolled. The incidences of hyponatremia (serum sodium level of＜135mEq/L) and severe hyponatremia (serum sodium level of＜130mEq/L) within the first 14 days after admission were 51ｵ (n＝42) and 20ｵ (n＝16), respectively. After admission, hyponatremia took a median period of 7 days to develop and lasted for a median of 3 days. Multivariate analysis demonstrated that higher fluid intake from days 1 to 3 and the presence of cranial fractures were risk factors for hyponatremia. The 58 patients with hyponatremia experienced fewer ventilator-free days, longer intensive care unit stays, and less favorable outcomes compared to the 24 patients without hyponatremia;however, these differences were not significant. Further studies are needed to determine the optimal management strategy for TBI-associated hyponatremia in the intensive care unit setting. No potential conflict of interest relevant to this article was reported.

日本薬学会 Acta Medica Okayama 00316903 128 3 2008 食後高血糖が血管反応性に及ぼす影響 419 424 EN Yoshito Zamami Shingo Takatori Yukiko Iwatani Kosuke Yamawaki Satoko Miyashita Nana Yabumae Fusako Takayama Mitsunobu Mio Hiromu Kawasaki Recent clinical studies demonstrated that transient postprandial hyperglycemia and hyperinsulinemia may contribute to the development of hypertension. Therefore, we investigated influence of acute hyperglycemia and/or hyperinsulinemia induced by glucose or insulin infusion on neuronal and humoral control of vascular tone in rats. Euglycemic male Wistar rats were pithed under anesthesia and arterial blood pressure was measured. Changes in vascular responses to spinal cord stimulation (SCS) and intravenous bolus injections of noradrenaline, angiotensin II, calcitonin generelated peptide (CGRP), acetylcholine and sodium nitroprusside (SNP) were studied by infusing various concentration of glucose or insulin. Continuous glucose infusion, which increased both blood glucose and serum insulin levels, significantly augmented adrenergic nerve-mediated pressor responses to SCS without affecting injection of pressor responses to noradrenaline or angiotensin II. In pithed rats with artificially increased blood pressure and blockade of autonomic outflow, glucose infusion attenuated CGRPergic nerve-depressor responses to SCS without affecting depressor responses to injection of CGRP, acetylcholine or SNP. In pithed rats treated with octreotide, which increased blood glucose without increasing serum insulin levels, glucose infusion caused only significant augmentation of adrenergic nervemediated pressor responses. Combined infusion of insulin and glucose, which resulted in increased serum insulin levels with euglycemic, significantly augmented adrenergic nerve-mediated pressor responses and attenuated CGRPergic nerve-mediated depressor responses. The present results suggest that acute hyperglycemia and hyperinsulinemia increases adrenergic nerve-mediated vasoconstriction, which is partly associated with the blunted CGRPergic nerve function, and that plasma insulin concentration associated with hyperglycemia may be responsible for alteration of neuronal vascular regulation. No potential conflict of interest relevant to this article was reported.

日本薬学会 Acta Medica Okayama 0031-6903 130 6 2010 Propolis 長期投与による Otsuka Long-Evans Tokushima Fatty (OLETF) ラットのインスリン抵抗性改善作用 833 840 EN Yoshito Zamami Hiroki Fujiwara Miho Hosoda Hayato Hino Kazuhiro Hirai Kazuaki Okamoto Hiromu Kawasaki Propolis is known to have abundant bioactive constituents and a variety of biological activities. To investigate the effect of Brazilian propolis on insulin resistance, 10-week-old Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a non-insulin-dependent type 2 diabetic model, were treated for 4 weeks with propolis (100 and 300 mg/kg, p.o.) or vehicle (control). Propolis treatment significantly decreased the plasma levels of insulin and insulin resistance index (Homeostasis Model Assessment-Insulin Resistance; HOM-IR), without affecting blood glucose levels and tended to lower systolic blood pressure compared with the control. In isolated and perfused mesenteric vascular beds of OLETF rats, propolis treatment resulted in significant reduction of sympathetic nerve-mediated vasoconstrictor response to periarterial nerve stimulation (PNS) and tended to increase calcitonin gene-related peptide (CGRP) nerve-mediated vasodilator response to PNS compared with in vehicle-treated OLETF rats. However, propolis treatment did not significantly affect the vasoconstrictor and vasodilator response to noradrenaline, CGRP, acetylcholine, and sodium nitroprusside. These results suggest that propolis could be an effective and functional food to prevent development of insulin resistance. No potential conflict of interest relevant to this article was reported.

岡山実験動物研究会 Acta Medica Okayama 31 2015 次亜塩素酸水溶液による環境消毒について 33 36 EN Koji Yamashita Yuji Hamamoto Yuto Yasuda Tomoko Ono Weak Acid Hypochlorous Solution (WAHS) has been used for disinfection of foods (meats, vegetables etc.), and for environmental disinfection in the Retirement homes, Hospitals and Laboratory Animal facilities. We will introduce here some of environmental disinfection tests. 1) To study whether WAHS is available or not for blood blot inoculated by Acinetobacter baumannii on the plate comparing with Sodium Hypochlorite. 2) Comparison of Ethanol and WAHS on the floor and handrail. 3) To study efficacy of shallowly dipping by WAHS on wagon caster inoculated by Staphylococcus aureus. The results are: 1) It was observed that WAHS had an efficacy equal to Hypochlorite with lower concentration in the blood test, but in case that the adhesion amount of blood was larger, much higher concentration or adding physical removal was needed. 2) Ethanol and WAHS had Equivalent efficacy on the test of floor and handrail. 3) It was suggested that shallowly dipping by WAHS was available for disinfection of wagon caster. We hope to proceed to confirm how to use WAHS for environmental disinfection. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2015 Long-Term Treatment with the Sodium Glucose Cotransporter 2 Inhibitor, Dapagliflozin, Ameliorates Glucose Homeostasis and Diabetic Nephropathy in db/db Mice EN Naoto Terami No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0360-4012 92 1 2014 Reducing Hemorrhagic Complication by Dabigatran Via Neurovascular Protection After Recanalization With Tissue Plasminogen Activator in Ischemic Stroke of Rat 46 53 EN Syoichiro Kono Kentaro Deguchi Yoshio Omote Taijun Yunoki Toru Yamashita Tomoko Kurata Yoshio Ikeda Koji Abe This study assesses the risks and benefits of tissue plasminogen activator (tPA) treatment under oral anticoagulation with dabigatran compared with warfarin or vehicle control in transient middle cerebral artery occlusion (tMCAO). After pretreatment with warfarin (0.2 mg/kg/day), dabigatran (20 mg/kg/day), or vehicle (0.5% carboxymethyl cellulose sodium salt) for 7 days, tMCAO was induced for 120 min, followed by reperfusion and tPA (10 mg/kg/10 ml). Clinical parameters, including cerebral infarction volume, hemorrhagic volume, and blood coagulation, were examined. At 24 hr after reperfusion, markers for the neurovascular unit at the peri-ischemic lesion were immunohistochemically examined in brain sections, and MMP-9 activity was measured by zymography. Paraparesis and intracerebral hemorrhage volume were significantly improved in the dabigatran-pretreated group compared with the warfarin-pretreated group. A marked dissociation between astrocyte foot processes and the basal lamina or pericyte was observed in the warfarin-pretreated group, which was greatly improved in the dabigatran-pretreated group. Furthermore, a remarkable activation of MMP-9 in the ipsilateral warfarin-pretreated rat brain was greatly reduced in dabigatran-pretreated rats. The present study reveals that the mechanism of intracerebral hemorrhage with warfarin-pretreatment plus tPA in ischemic stroke rats is the dissociation of the neurovascular unit, including the pericyte. Neurovascular protection by dabigatran, which was first shown in this study, could partially explain the reduction in hemorrhagic complication by dabigatran reported from clinical study. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0013-9580 52 5 2011 Therapy for hyperthermia-induced seizures in Scn1a mutant rats 1010 1017 EN Keiichiro Hayashi Satoshi Ueshima Mamoru Ouchida Tomoji Mashimo Teiichi Nishiki Toshiaki Sendo Tadao Serikawa Hideki Matsui Iori Ohmori Purpose: Mutations in the SCN1A gene, which encodes the alpha 1 subunit of voltage-gated sodium channels, cause generalized epilepsy with febrile seizures plus (GEFS+) and severe myoclonic epilepsy of infancy (SMEI). N1417H-Scn1a mutant rats are considered to be an animal model of human FS+ or GEFS+. To assess the pharmacologic validity of this model, we compared the efficacies of eight different antiepileptic drugs (AEDs) for the treatment of hyperthermia-induced seizures using N1417H-Scn1a mutant rats. Methods: AEDs used in this study included valproate, carbamazepine (CBZ), phenobarbital, gabapentin, acetazolamide, diazepam (DZP), topiramate, and potassium bromide (KBr). The effects of these AEDs were evaluated using the hot water model, which is a model of experimental FS. Five-week-old rats were pretreated with each AED and immersed in water at 45 degrees C to induce hyperthermia-induced seizures. The seizure manifestations and video-electroencephalographic recordings were evaluated. Furthermore, the effects of each AED on motor coordination and balance were assessed using the balance-beam test. Key Findings: KBr significantly reduced seizure durations, and its anticonvulsant effects were comparable to those of DZP. On the other hand, CBZ decreased the seizure threshold. In addition, DZP and not KBr showed significant impairment in motor coordination and balance. Significance: DZP and KBr showed potent inhibitory effects against hyperthermia-induced seizures in the Scn1a mutant rats, whereas CBZ exhibited adverse effects. These responses to hyperthermia-induced seizures were similar to those in patients with GEFS+ and SMEI. N1417H-Scn1a mutant rats may, therefore, be useful for testing the efficacy of new AEDs against FS in GEFS+ and SMEI patients. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 2014 Study on anti-oxidative and anti-inflammatory effects of radon inhalation on mouse models of oxidative stress-related disease 第１編　Suppression of streptozotocin-induced type-1 diabetes in mice by radon inhalation 第２編　Suppression of dextran sulfate sodium-induced colitis in mice by radon inhalation EN Yuichi Nishiyama No potential conflict of interest relevant to this article was reported.

岡山実験動物研究会 Acta Medica Okayama 30 2014 筋ジストロフィーニワトリの水分代謝異常 7 7 EN Noboru Saito The muscular dystrophy chicken has been studying as model animal of muscular dystrophy for more than 50 years. Recently, the mutation of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) gene has been identified as a responsible for muscular dystrophy chicken. We observed that muscular dystrophy chicken not only showed the degeneration of skeletal muscles but also produced watery feces. Therefore, we examined the possibility of abnormalities in water metabolism of muscular dystrophy chicken. We first analyzed plasma osmolality and gene expression of aquaporin 2 (AQP2), AQP3 and alpha subunit of the amiloride-sensitive epithelial sodium channel (αENaC) in muscular dystrophy chicken and White Leghorn chicken under normal physiological conditions at five-week old. Subsequently, we analyzed these same parameters after one-day water-deprivation. The main findings of our study are that: I) the plasma osmolality was significantly higher in muscular dystrophic chicken than in White Leghorn; II) kidney αENaC mRNA expression was significantly lower in muscular dystrophic chicken than in White Leghorn; III) AQP2 and AQP3 mRNA expressions in muscular dystrophic chicken were similar in White Leghorn. We suggest that the mutation of WWP1 may cause the abnormality of sodium absorption, and thus muscular dystrophic chicken become hypernatremic. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0306-4522 235 2013 EFFECT OF LIDOCAINE ON DYNAMIC CHANGES IN CORTICAL REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE FLUORESCENCE DURING TRANSIENT FOCAL CEREBRAL ISCHEMIA IN RATS 59 69 EN H. Naito Y. Takeda T. Danura I. S. Kass K. Morita Rats were subjected to 90 min of focal ischemia by occluding the left middle cerebral and both common carotid arteries. The dynamic changes in the formation of brain ischemic areas were analyzed by measuring the direct current (DC) potential and reduced nicotinamide adenine dinucleotide (NADH) fluorescence with ultraviolet irradiation. In the lidocaine group (n = 10), 30 min before ischemia, an intravenous bolus (1.5 mg/kg) of lidocaine was administered, followed by a continuous infusion (2 mg/kg/h) for 150 min. In the control group (n = 10), an equivalent amount of saline was administered. Following the initiation of ischemia, an area of high-intensity NADH fluorescence rapidly developed in the middle cerebral artery territory in both groups and the DC potential in this area showed ischemic depolarization. An increase in NADH fluorescence closely correlated with the DC depolarization. The blood flow in the marginal zone of both groups showed a similar decrease. Five minutes after the onset of ischemia, the area of high-intensity NADH fluorescence was significantly smaller in the lidocaine group (67% of the control; P = 0.01). This was likely due to the suppression of ischemic depolarization by blockage of voltage-dependent sodium channels with lidocaine. Although lidocaine administration did not attenuate the number of pen-infarct depolarizations during ischemia, the high-intensity area and infarct volume were significantly smaller in the lidocaine group both at the end of ischemia (78% of the control; P = 0.046) and 24 h later (P = 0.02). A logistic regression analysis demonstrated a relationship between the duration of ischemic depolarization and histologic damage and revealed that lidocaine administration did not attenuate neuronal damage when the duration of depolarization was identical. These findings indicate that the mechanism by which lidocaine decreases infarct volume is primarily through a reduction of the brain area undergoing NADH fluorescence increases which closely correlates with depolarization. No potential conflict of interest relevant to this article was reported.

Informa Healthcare Acta Medica Okayama 1071-5762 47 3 2013 Inhibition of arginase ameliorates experimental ulcerative colitis in mice 137 145 EN Y. Akazawa M. Kubo R. Zhang K. Matsumoto F. Yan H. Setiawan H. Takahashi Y. Fujikura K. Ogino Nitric oxide (NO) is produced from the conversion of L-arginine by NO synthase (NOS) and regulates a variety of processes in the gastrointestinal tract. Considering the increased activity of arginase in colitis tissue, it is speculated that arginase could inhibit NO synthesis by competing for the same L-arginine substrate, resulting in the exacerbation of colitis. We examined the role of arginase and its relationship to NO metabolism in dextran sulfate sodium (DSS)-induced colitis. Experimental colitis was induced in mice by administration of 2.5% DSS in drinking water for 8 days. Treatment for arginase inhibition was done by once daily intraperitoneal injection of N-omega-hydroxy-norarginine (nor-NOHA). On day 8, we evaluated clinical parameters (body weight, disease activity index, and colon length), histological features, the activity and expression of arginase, L-arginine content, the expression of NO synthase (NOS), and the concentration of NO end-product (NOx: nitrite + nitrate). Administration of nor-NOHA improved the worsened clinical parameters and histological features in DSS-induced colitis. Treatment with nor-NOHA attenuated the increased activity of arginase, upregulation of arginase. at both mRNA and protein levels, and decreased the content of L-arginine in colonic tissue in the DSS-treated mice. Conversely, despite the decreased expression of NOS2 mRNA, the decreased concentration of NOx in colonic tissues was restored to almost normal levels. The consumption of L-arginine by arginase could lead to decreased production of NO from NOS, contributing to the pathogenesis of the colonic inflammation; thus, arginase inhibition might be effective for improving colitis. No potential conflict of interest relevant to this article was reported.

RSC Publishing Acta Medica Okayama 1759-9660 5 11 2013 Determination of polyamines in Arabidopsis thaliana by capillary electrophoresis using salicylaldehyde-5-sulfonate as a derivatizing reagent 2854 2859 EN Genki Inoue Takashi Kaneta Toshio Takayanagi Junichi Kakehi Hiroyasu Motose Taku Takahashi Herein, we report a novel method for the determination of polyamines in a sample extracted from Arabidopsis thaliana by capillary electrophoresis (CE) using salicylaldehyde-5-sulfonate (SAS) as a derivatizing reagent. An aldehyde group of SAS forms a Schiff base with amino groups of aliphatic polyamines, resulting in an anionic species with an absorption band in the ultraviolet region. The derivatization method was straightforward since the derivatives were formed by mixing a sample with the derivatizing reagent at a neutral pH. In addition, the negative charges induced by SAS led to a high resolution with a short analysis time. This method permitted the separation of five polyamines, which play important roles in plants. However, further improvement in sensitivity was needed for the determination of the polyamines in plant samples. Therefore, the CE method was coupled with solid-phase extraction (SPE) using an ion-pairing formation with sodium dodecyl benzene sulfonate. The SPE method improved the concentration limits of detection to sub-μM levels, which corresponded with a 10-fold enhancement. The calibration curves for cadaverine, putrescine, and spermidine were linear with concentrations that ranged from 1 to 20 μM and correlation coefficients (R2) were greater than 0.998. The proposed method was applied to the determination of spermidine in a plant sample, Arabidopsis thaliana. No potential conflict of interest relevant to this article was reported.

Ohara Instituts für landwirtschaftliche Biologie Acta Medica Okayama 16 1 1973 THE ROLE OF CALCIUM IN SELECTIVE CATION UPTAKE BY PLANT ROOTS II. Effects of Temperature，Desorption Treatment and Sodium Salt on Rubidium Uptake* 19 28 EN Toshio Kawasaki Shiro Hori No potential conflict of interest relevant to this article was reported.

岡山大学農学部 Acta Medica Okayama 2186-7755 102 2013 日長，補光強度，遮光時期および最低夜温がシュッコンカスミソウ ‘アルタイル’の形態異常花序発生に及ぼす影響 29 34 EN Norihito Yamaguchi Tanjuro Goto Kayoko Kobiki Shoko Otani Yuichi Yoshida As occurrence of abnormal inflorescence in Gypsophila paniculata ‘Altair’ is caused by environmental conditions, effects of day length, supplemental lighting strength, shading period and minimum night temperature on occurrence of abnormal inflorescence were investigated. Abnormal inflorescence was classified into four types : normal, pattern 1 (Short-flower stalk), pattern 2 (Coalescent two-flower stalk) and pattern 3 (Looping and irregular-flower stalk). Neither of 12h, 16h, 20h or 24h day length by fluorescent lamp, nor 24h by incandescent lamp affected occurrence of abnormal inflorescence. Effects of four levels of light intensity (fluorescent lamp : PPFD 1μmol・m－2・s－1, incandescent lamp : PPFD 3μmol・m－2・s－1, metal halide lamp : PPFD 14μmol・m－2・s－1 and high-pressure sodium lamp : PPFD 48μmol・m－2・s－1) were examined in 16h photoperiod. Occurrence of abnormal inflorescence was not affected by different light intensities, neither was it affected by shading period. Occurrence of abnormal inflorescence at 15°C was however significantly reduced compared to that at 8°C. In particular, patterns 2 and 3 at 15°C were significantly reduced compared to those at 8°C. There was a strong negative correlation between average night temperature from starting the treatment to flower budding (7.1°C, 9.0°C, 9.2°C, 11.6°C and 16.4°C) and incidence of pattern 3 (13.1%, 8.7%, 7.1%, 1.1% and 0.7%). Therefore, as average night temperature increased, occurrence of abnormal inflorescence decreased. The results show that low night temperature may be the main factor inducing occurrence of abnormal inflorescence. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 0378-7753 225 2013 NMR study for electrochemically inserted Na in hard carbon electrode of sodium ion battery 137 140 EN Kazuma Gotoh Toru Ishikawa Saori Shimadzu Naoaki Yabuuchi Shinichi Komaba Kazuyuki Takeda Atsushi Goto Kenzo Deguchi Shinobu Ohki Kenjiro Hashi Tadashi Shimizu Hiroyuki Ishida The state of sodium inserted in the hard carbon electrode of a sodium ion battery having practical cyclability was investigated using solid state 23Na NMR. The spectra of carbon samples charged (reduced) above 50 mAh g－1 showed clear three components. Two peaks at 9.9 ppm and 5.2 ppm were ascribed to reversible sodium stored between disordered graphene sheets in hard carbon because the shift of the peaks was invariable with changing strength of external magnetic field. One broad signal at about －9 to －16 ppm was assigned to sodium in heterogeneously distributed closed nanopores in hard carbon. Low temperature 23Na static and magic angle spinning NMR spectra didn't split or shift whereas the spectral pattern of 7Li NMR for lithium-inserted hard carbon changes depending on the temperature. This strongly suggests that the exchange of sodium atoms between different sites in hard carbon is slow. These studies show that sodium doesn't form quasi-metallic clusters in closed nanopores of hard carbon although sodium assembles at nanopores while the cell is electrochemically charged. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 66 5 2012 Dravet Syndrome : A Genetic Epileptic Disorder 369 376 EN Mari Akiyama Katsuhiro Kobayashi Yoko Ohtsuka Review 10.18926/AMO/48961 Dravet syndrome (DS), or severe myoclonic epilepsy in infancy, is one of the most severe types of genetic epilepsy. It is characterized by the initial occurrence of febrile or afebrile seizures that often evolve into status epilepticus in infants with normal development, and by the subsequent appearance of myoclonic and/or atypical absence seizures as well as complex partial seizures. The key feature that characterizes DS is fever sensitivity, although photosensitivity and pattern-sensitivity are also often seen. The prognosis is unfavorable in most cases. Seizures become drug-resistant and persist, with many patients suffering from motor and cognitive impairment. Mutations of SCN1A, which encodes the voltage-gated sodium channel NaV1.1, are the most frequent genetic cause of this syndrome. SCN1A mutations and/or microchromosomal rearrangements involving SCN1A are detected in about 85% of patients. Mutations of PCDH19 have also been reported in female patients with clinical findings compatible with DS. PCDH19 mutations might account for 5% of overall DS cases. Thirty years after its first description, DS is considered as a model of channelopathy. This survey reviews recent developments in the research literature on DS, focusing on the clinical course, as well as its genetic causes. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 1881-0942 82 2 2011 An Efficient Synthesis of Antibiotic SF-2312 (3-Dihydroxyphosphoryl-1,5-dihydroxy-2-pyrrolidone) 1675 1683 EN Tadashi Hanaya Chika Itoh N-Benzyloxy-2-(diethoxyphosphoryl)pent-4-enamide (6) was prepared from ethyl diethoxyphosphorylacetate in a 3-step sequence. Oxidative cleavage of the terminal olefin of 6 with osmium tetroxide and sodium periodate afforded 1-benzyloxy-3-diethoxyphosphoryl-5-hydroxy-2-pyrrolidone (7). The first synthesis of racemic SF-2312 was achieved by treatment of 7 with trimethylsilyl bromide, followed by hydrogenolysis. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 124 2 2012 Scn1a遺伝子変異ラットを用いた抗てんかん薬の熱性けいれん抑制効果 115 117 EN Keiichiro Hayashi Iori Ohmori Hideki Matsui No potential conflict of interest relevant to this article was reported.

岡山大学農学部 Acta Medica Okayama 2186-7755 101 2012 根損傷の軽減が塩土壌における移植イネの生育と乾物重におよぼす影響 7 12 EN Makoto Tsuda Yukie Ueda Yoshihiko Hirai Rice root has an important role in water absorption and exclusion of sodium ion in saline soil. However, it is injured during transplanting to paddy field. The objective of this study was to examine whether prevention of such root injury reduces salinity damage in rice. Extremely early rice cultivar "Akihikari" that is not salt tolerant was grown in plastic bags filled with soil. Roots of seedlings were clipped leaving 5mm~10mm, then transplanted to 5L pots filled with soil applied with 0g, 7g and 10g of sodium chloride (NaCl). The growth and dry weight at the mature stage was compared to those of plants transplanted with intact roots. Time of transplanting was 12, 19, 26 and 33 days after sowing. Increase of number of stems was suppressed due to root clipping and salinity, but there was no interaction between the two factors. Dry weight of total above ground part and panicle decreased due to salinity though root clipping did not change them. The reduction of panicle dry weight was caused by the reduction of mean number of spikelets per panicle, thus mean panicle dry weight, as number of panicles and mean spikelet dry weight showed no changes. It was concluded that prevention of the root injury may not effectively alleviate salinity damage in less salt tolerant rice cultivars. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 123 3 2011 生活習慣病治療のパラダイムシフト―慢性炎症を標的とした治療戦略― 197 206 EN Kenichi Shikata No potential conflict of interest relevant to this article was reported.

The Ceramic Society of Japan Acta Medica Okayama 1882-0743 118 1379 2010 Phase separation of borosilicate glass containing sulfur 603 607 EN Keiji Saiki Shinichi Sakida Yasuhiko Benino Tokuro Nanba A 10Na(2)S·30B(2)O(3)·60SiO(2) (mol %) glass was prepared, and the changes in glass structure and chemical state of sulfur caused by phase separation were investigated. In the as-prepared and heat-treated glasses, sulfur was present as S(2)－ anion and polysulfide S(2)－ and S(3)－ anions, and Si–S and B–S bonds were not confirmed. A phase separation by spinodal decomposition was observed after heat-treatment, where sulfur was preferentially distributed to borate-rich phase. Even after the phase separation, formation of non-bridging oxygen was not recognized. The preferential distribution of sulfur anions in the present glass was explainable on the basis of the change in population of sodium ions, which compensated the negatively-charged sulfur anions. No potential conflict of interest relevant to this article was reported.

Springer Japan Acta Medica Okayama 1431-0651 15 3 2011 Characterization of an OmpA-like outer membrane protein of the acidophilic iron-oxidizing bacterium, Acidithiobacillus ferrooxidans 403 410 EN Mohammed Abul Manchur Mei Kikumoto Tadayoshi Kanao Jun Takada Kazuo Kamimura An OmpA family protein (FopA) previously reported as one of the major outer membrane proteins of an acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans was characterized with emphasis on the modification by heat and the interaction with peptidoglycan. A 30-kDa band corresponding to the FopA protein was detected in outer membrane proteins extracted at 75A degrees C or heated to 100A degrees C for 10 min prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). However, the band was not detected in outer membrane proteins extracted at a parts per thousand currency sign40A degrees C and without boiling prior to electrophoresis. By Western blot analysis using the polyclonal antibody against the recombinant FopA, FopA was detected as bands with apparent molecular masses of 30 and 90 kDa, suggesting that FopA existed as an oligomeric form in the outer membrane of A. ferrooxidans. Although the fopA gene with a sequence encoding the signal peptide was successfully expressed in the outer membrane of Escherichia coli, the recombinant FopA existed as a monomer in the outer membrane of E. coli. FopA was detected in peptidoglycan-associated proteins from A. ferrooxidans. The recombinant FopA also showed the peptidoglycan-binding activity. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 123 2 2011 成人期に達したDravet症候群の長期経過に関する研究 91 95 EN Mari Akiyama Katsuhiro Kobayashi Harumi Yoshinaga Yoko Ohtsuka No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 40 4 1928 On the action of some diuretica and other drugs upon the glomeruli 717 726 EN Masanori Okada From the results of my experiments the conclusion may be summarized as follows. 1. Caffeine, caffeine sodio-benzoate and diuretin have dilator effect upon the glomerular capillaries and the circulation in them becomes more rapid. The quantity of blood which flows through them increases and a number of glomeruli, hitherto unobserved, become visible. On the other hand, theocin and glucose have no effect upon the glomeruli. 2. Solution of potassium acetate causes a remarkable contraction and at the same time the glomeruli till then observed, become invisible. 3. Kreatin produces a dilatation of glomerular capillaries and accordingly the circulation in them becomes more rapid and also the number of glomeruli increased. Guanidin in a solution weaker than 0.4 p. c. causes dilatation of glomerular capillaries and increase of the quantity of blood which flows through them and also it causes the increase of verocity of blood flow in the glomerular capillaries. The glomeruli, hitherto stationary, become active and the number of them increases. On the contrary, a solution stronger than 0.5 p. c. always causes contraction of glomerular capillaries and the number of glomeruli are decreased. 4. Isotonic solution (with 0.6% Ringer's solution) of NaNO(2), NaNO(3) and Na(2)HPO(4) causes a slight dilatation of glomerular capillaries and accordingly the increase of verocity of blood flow in them and of quantity of blood flowing through them are produced. But other sodium salts have no effect upon the glomeruli. All of potassium salts, on the contrary, have an opposite effect upon the glomeruli to that of sodium salts. 5. Calcium salts and magnesium salts (isotonic with 0.6% Ringer's solution) and some organic substances (m/100 in density) have a remarkable dilator effect upou the glomerular capillaries and cause the increase of verocity of blood flow in them and of quantity of blood which flows through them. Glomeruli are increased in number und are remarkably congested and swollen. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 44 5 1932 「オルニツール」酸ノ藥理學的作用ニ就テ 1530 1552 EN Mitsutaka Nakano The pharmacological action of phenol-glycuronic acid, urochloralic acid, glycuronic acid and benzoyl-glycuronic acid have been investigated in our Laboratory by Ishii, Matsushima, Tanaka and Sakata. Following the line of these investigations the author has studied the pharmacological action of ornithuric acid prepared from the urine of hen by giving sodium benzoate. The results obtained are as follows: (1) Ornithuric acid when given to the rabbit in its general circulation gives rise to a marked diuresis. Meanwhile it causes practically no change in general blood pressure, nor in the water content of the blood, and only the oncometric record shows an augmentation of kidney volume in every case. And in denervated or isolated kidney an increased perfusion rate is obtained. So it seems that in this vasomotor reaction the central nervous mechanism plays no part. (2) Ornithuric acid exerts diuretic action also on cold blooded animal. Analysis effected on perfused toad kidney showed that it concerns in this diuresis the glomerular system only, and that tubular system has nothing to do with. (3) The action of ornithuric acid on the blood vessel of the organs other than kidney is as follows: - Hind leg vascular system of the frog is dilated by low and constricted by high concentration of this substance in the perfusing fluid. Vascular system of the rabbit's ear shows similar reaction toward this substance. But coronary vascular system is constricted notwithstanding the concentration of this substance in the perfusing fluid. (4) Isolated hearts of toad and rabbit augment their tone and diminish the amplitude of their contraction by middle dosage and especially in the beginning, but then tone returns gradually to normal, accompanying an increase in the amplitude of contraction. By larger dosages there occurs an increase in amplitude of the heart beat during only a very short period at the beginning, then the amplitude decreases progressively to be finally arrested at systole. It must be noted that in every case there is no change in the frequency of heart beat. (5) In isolated smooth muscular organs of the rabbit, such as intestine, uterus and bladder, an augmentation in the tone and the spontaneous movement are observed for an appropriate dosage. Atropine has no influence on this effect, also the isolated trigonal and basal parts of the rabbit's bladder have shown no difference in their reaction toward this substance. (6) From these upper three results it may be concluded that ornithuric acid does not exert its action by way of the nervous mechanism but directly on the smooth muscular system. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 46 12 1934 實驗的家兎佝僂病ニ對スル「トレンタール」ノ作用ニ就テ 3065 3073 EN Masanobu Okumura Experiments were undertaken to determine the value of Trental, a sodium salt of dimethylamino-methylphenyl phosphoric acid, in rickets of young growing rabbits. The standard rickets-producing dietary was fed and, as a supplement, Trental was given subcutaneously. All the control animals which were fed the rickets-producing diet, without exception developed rickets. In the large series of animals received Trental developed rickets in somewhat slighter degree and showed a slightly better increase of both body weight and bone length than that of the control animals, although in the small series of the animals developed rickets to the same extent as the control animals. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 47 4 1935 電流ノ生物ニ對スル作用ニ就テ（第3報）電流刺戟ニヨル細胞膜ノ透過性ノ變化ニ就テ 1069 1093 EN Nobutane Mori The auther studied on alternation of permeability of cell-membranes by electric current and reached to the following results. 1) Plasmolysis of epithelial cells of Tradescantia virginica by hypertonic solution (0.5-1.0%) of sodium chloride will be stopped at the neighbourhood of the cathode by passing through of the electric current. 2) In the neighbourhood of cathode potassium-ions escape from the cells into the surrounding medium. It will be so understanded that the damage of semipermeability of the cell-membranes allows passage of anion and potassium-ion escapes in balance of it. Same thing occurs on the muscle cutaneus dorsi of frog. 3) Potassium-escape can be observed on the resting muscle of frog when the temperature of bathing fluid exceeds 29-32°C. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 52 7 1940 各種植物及ビ2, 3藥物ノ利尿作用ニ就テ（第1報）「商陸エキス」ノ利尿作用ニ就テ 1664 1674 EN Hiromu Masuzawa By the perfusion method through the kidney of the toad following results were obtained. 1) When the Extr. of Rad. Phytol. is added to the perfusing fluid, it increases distinctly the flow of urine. 2) Conversely, the addition of potassium salt, which will be contained abundantly in Rad. Phytol., brings, firstly, the remarkable decrease of the urine flow, and then it may increase slightly. 3) Ash of the Rad. Phytol. dissolved in Ringer's solution decreases the urine flow remarkably at first, and then it may increase. 4) When the Extr. of Rad. Phytol. is applied in drops to the exposed kidney of frog, causes dilatation of glomerular capillaries; the circulation in them becomes more rapid, i.e. the quantity of blood which flows through them increases, and a number of glomeruli, hitherto invisible, becomes visible. The application of this extract in drops on the web of frog dilates its blood-capillaries and increases the blood-flows through them. 5) But the solution of potassium salt used, or the ash of the Rad. Phytol. causes contraction of glomerular capillaries and capillaries of web of frog. The blood-flows in the capillaries become remarkably slow, i.e. the quantity of blood which flows through them decreases. The glomeruli, hitherto activ, becomes stationary and visible number of glomeruli diminishes. 6) The measurement of the freezing-point revealed that the osmotic pressure of the Extr. of Rad. Phytol. used was not very different from the normal Ringer's solution. 7) By the flame-reaction it was ascertained that potassium and sodium salts were contained in the Rad. Phytol. 8) Diuretic action of Rad. Phytol. is due to some organic substance, which causes dilatation of capillaries of glomerulus and web of frog. Potassium salt which is contained in Rad. Phytol. makes some additional action through hydraemia. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 54 8 1942 Acetylcholin及ビ抗Cholinesterase劑ノ作用機轉補遺（其ノ3）諸種麻醉藥ノCholinesteraseニ對スル作用附諸種麻醉藥ノ腦髓竝ニ脊髓A.Ch.含有量ニ及ボス影響 1430 1448 EN Kazumasa Okuda The author had studied the action of some narcotics (urethane, chloralhydrate, barbital, luminal-Na, sulfonal, morphine hydrochl., ether and chloroform) on choline esterase and the influence of these on acetylcholine content in the central nervous system and the result is as follows: 1) These drugs inhibited the action of choline esterase. 2) The intensity of the action of these drugs (except ether and chroloform) on esterase had following order, i.e. morphine>barbital>chloralhydrate>sulfonal> urethaue>luminal. Except of sodium salt of luminal the action of those drugs is approximately proportional to their molecular weight. 3) The relation of the inhibitory action between these drugs (added luminal, brovarin) and choline esterase is linear. 4) The acetylcholine content in the central nervous system of the mouse was increased by these drugs. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 55 3 1943 微量物質ノ比重測定1新法 429 436 EN Takamaro Gesi The method here described is an improvement of that developed by LinderstrømLang, Lanz and Jacobsen (1938, 1940). A reasonably linear specific gravity gradient is produced in a vertical measuring cylinder by mixing kerosene and bromobenzene, or benzene and chroloform in varying proportions. In kerosene-bromobenzene mixtures the position of a drop is determined after the drop (diameter 2.0mm) has fallen in 30 sec. Plotting the positions of drops of known solutions of sodium chloride as ordinates against the corresponding specific gravities as abscissae, a nearly straight line is obtained in a coordinate system, from which it is possible, knowing the position of the drop of the unknown solution, to read the specific gravity of the solution with considerable accuracy. The reading of the positions of drops may be done with the naked eye making use of two sheeves of section papers, each of which are stick on at the diametral wall of measuring cylinder. The eye is kept at the level of the drop and the same position of two scale divisions, in order to eliminate errors due to parallaxis. In benzene-chloroform mixtures the drops fall more sapid, and come to rest much faster, so that the infusion of the mixtures into the drops will be minimized. At the constant temperature it is not advisable to use of kerosene-bromobenzene mixtures for more than about 1 hour at one experiment, and to use of benzene-chloroform mixtures for more than about 2 hours. Therefore, benzene-chloroform mixtures are more suitable than kerosene-bromobenzene mixtures for the determination of specific gravity. Its accuracy is 0.1 per cent. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 66 5 1954 大腸菌ファージの実験的研究第一編大腸菌ファージのクロマトグラフ 909 918 EN K. Sasaki Since 1940 the study of phage opened not only the beginning of co-operate study for physicists and chemists, but also the phage came to be clearified its character as the Bacterio Virus gradually. Then the study of phage made not a few contributions to the development of the study of Virus as model material of its kind. Now, these results are due to the endeavors of such scholars as Delbrück, Anderson, Cohen, Putznam, Doermann, Wyckoff and their co-operators mainly. Then, Charles. C. Shepard found out the characteristics of a distinctive nature in paperstrips in the course of investigations for the adsorptive behavior of Viruses. I tried some experiments in the paperchromatograph and capillarglass-chromatograph about the conditions that the phagef of coli-strains being adsorbed and recovered by paper and capillarglass. 1. The solvent employed here was 0.1% bovine serum albumin in 0.1 M sodium chloride. 2. The solution employed was colibacteriophage diluted appropriately in the same solvent. 3. The adsorbent employed was Toyo-filterpaper and capillarglass which were treated previously by 0.01 M hydrochloric acid and then washed by glasswather. 4. The experiments were carried out in an ice-box. At the beginning of an experiment a paperstrip was placed in the test tube so that it might not touch the sides, but touch the 20 ml. of solvent placed beforehand in the bottom of the tube. When the water front passed the second 1.25 cm section, 0.002 ml. of phage diluted in the same solvent appropriately as was placed in the bottom of the tube was quickly placed in the center of the second section and. the paper returned to the position in the tube. When the water front passed the last marked section, the paper was removed from the tube and ten marked sections were quickly cut off into their respective ten test tubes each containing of a culture of E. coli-strains. For a period of not more than 10 minutes, the ten sections of paper were stirred occasionally and pressed against the sides of the tubes. At the end of it, 0.5 ml. were taken out and added it to 3.0 ml. of 0.7% agar; the resultant 3.5 ml. were mixed up and poured on a 1%, agarplate. There was recovered viable phage in a considerable degree. 5. It seems to me that this recoveries of phage has been carried out due to the presence of bovine serum albumin. The chromatograph conducted here would seem to involbe the mixture of elution and displacement in the Terminology of Tiselius. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 10 1957 ブドウ球菌のグルタミン酸呼吸に対する諸種阻害剤及び抗生物質の作用 2505 2510 EN Yoshiro Yabe Kazuo Akita Yoshimi Akita The author studied the action of various inhibitors and antibiotics on the glutamaterespiration of staphylococci under the consideration of physiological structure of the cell. Staphylococcus citreus and aureus (Terashima) were used as the test organisms, and L-glutamic acid as the substrate. The results were as follows: 1) Thionine, sodium arsenite and 8-hydroxyquinoline inhibited the respiration of intact cells of staphylococci markedly, but did not inhibit that of cell-free extracts. The inhibitive action of these three sorts of inhibitors was stronger on Staph. aureus (Terashima) than on Staph. citreus. 2) In intact cells, the inhibition of respiration by potassium cyanide and sodium azide was not restored by addition of thionine. In cell-free extracts, however, the inhibition by these two sorts of inhibitors was well restored by thionine. 3) By addition of thionine, the inhibition of the respiration of cell-free extracts by octanole was not so well restored as that by cyanide or azide. 4) 2, 4-Dinitrophenol inhibited the glntamate-respiration of both of the intact cells and cell-free extracts. 5) Of all the antibiotics tested, aureomycin was the only one which noticeably inhibited the glutamate-respiration of staphylococci. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 7 1957 慢性脳局所アナフィラキシー家兎脳髄の解糖作用ならびに組織呼吸に関する研究第3編慢性脳局所アナフィラキシー家兎脳髄の灌流実験法による糖代謝に関する研究 1765 1771 EN Yoshio Obo The glycolysis of the living brain of deacapitated chronic cerebral local anaphylactic rabbits was investigated by irrigation method. Glucose was added to the irrigating fluid and the consumption of glucose was measured. In the chronic cerebral local anaphylactic rabbits the glucose-consumption was markedly strained in comparison with that in normal group. The pyruvic acid increased in both anaphylactic and normal groups, but the anaphylactic group showed a less increase than the normal. The addition of sodium pyruvate to the irrigating fluid caused less decrease of pyruvic acid in chronic cerebral anaphylactic rabbits than that in normal group From the facts above mentioned it is considred, that glycolysis is strained in the brain of chronic erebral local anaphylactic rabbits. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 3 1957 パラチオン中毒に関する実験的並びに臨床的研究第三編パラチオン中毒治療剤と血清蛋白活性SH基及びムコ蛋白 947 958 EN Arimitsu Hukuhara SH medical compounds used in this study were BAL, sodium thiosulfate, cystine, menthionine and sodium mercaptoacetate. 1. In the animal tests on the effectiveness of SH medical compounds, BAL was the most effective and then sodium thiosulfate was next, while scarecely effect was observed in the experiments of cystine, methionine and sodium mercaptoacetate. 2. By the administration of PAM, SH enzyme such as mucoprotein was promptly and certainly reactivated as well as cholinesterase. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 2 1957 細菌のグルタミン酸代謝に関する研究第二篇白色ブドウ球菌のグルタミン酸代謝 549 560 EN Yoshimi Akita As to the physiological or metabolic features of pathogenic staphylococci, there are only a few reports, compared with those of Escherichia coli which was treated in report I. In this report, the author reports about the physiological aspects, particularly about the terminal respiratory system of Staphylococcus albus, entering from the studies of glutamic acid metabolism: 1) Staph. albus has the so-called citric acid cycle as its terminal respiratory ststem. 2) As a result of oxidative deamination, glutamic acid enters into the citric acid cycle and is further oxidized through this cycle. Glutamic acid is, however, best oxidized of all the intermediates of citric acid cycle and the related compounds. 3) Glutamic-aspartic and glutamic-alanic transaminations are carried out by this organism, in which glutamic acid plays the central role. 4) Divalent metal ions (Mg(++), Mn(++) and Fe(++)) show no remarkable effect on the glutamate-respiration of Staph. albus. 5) Of the various inhibitors tested, sodium azide, 2: 4-dinitrophenol, sodium arsenite and 8-hydroxyquinoline inhibit the glutamate-respiration strongly, and the most remarkable is the inhibitive action of 8-hydroxyquinoline. 6) Of the various antibiotics used, the inhibitive action of aureomycin is the most remarkable. Penicillin also shows some inhibitive action at pH 5.4. 7) The inhibition of the glutamate-respiration of this organism by these various inhibitors and antibiotics shows usually the tendency to rise up in the region of lower pH. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 2 1957 全身麻酔の生体防禦機能に及ぼす影響に関する研究第3編各種全身麻醉の血清殺菌力に及ぼす影響に関する研究 371 377 EN Shinobu Yumiyama 1) The basal narcosis caused no change of the serum sterilizing power. 2) In the intravenous anesthesia using penthothal sodium the sterilizing power showed a temporal decline but after the end of anesthesia it slightly increased. 3) In the endotracheal anesthsia with cyclopropaine, the sterilizing power always showed a marked decline. 4) In the endotracheal anesthsia with ether, the sterilizing power showed a temporal decline during and after anesthesia. 5) In the endotracheal anesthesia using cyclopropaine and nitrous oxide or with additional ether together, the serum sterilizing power varied as well as in that using cyclopropaine only, but did not decline so distinctly. 6) The variation of sterilizing power in the endotracheal anesthesia using nitrous oxide and ether together showed a very gradual decline and then also gradually recovered. 7) The variation of the sterilizing power caused by oxygen inhalation showed clear increment with the beginn of inhalation namely the oxygen inhalation is very important for protecting sterilizng power of serum from inhibition of it. 8) The influence of various general anesthesia upon the serum sterilizing power was independent from tne Opsonin rate in the blood coagulating time and the number of platelets. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 2 1957 全身麻醉の生体防禦機能に及ぼす影響に関する研究第2編各種全身麻醉の血中オプソニン率に及ぼす影響に関する研究 359 369 EN Shinobu Yumiyama The rise and fall of the Opsonine rate in the blood during various general anesthesia was investigated. 1) The basal narcosis scarsely gave influences upon the Opsonin rate. 2) During the intravenous anesthesia using Penthothal sodium the Opsonin rate fell to the bottom at the second hour of anesthesia. 3) During the endotracheal anesthesia with cyclopropaine the Opsonine rate showed its least value also at the second hour. 4) During the endotracheal anesthesia with ether, the Opsonin rate showed a gradule and marked increase with the progress of anesthesia. 5) During the endotracheal anesthesia using cyclopropaine and nitrous oxide together the Opsonin rate decreased markedly and after hours it showed the least value, then began to increase from the third day after anesthesia and on the 7th and 14th days, it recovered to the rate before the anesthesia. 6) During the endotracheal anesthesia using the combination of cyclopropaine, nitrous oxide and ether, the Opsonin rate decreased markedly at the first hour of the anesthesia then considerably increased at the second hour but considerably decreased at the third hour. And it gradually increased from the third day after the anesthesia. 7) During the endotracheal anesthesia using nitrous oxide and ether, it decreased markedly at the first hour of anesthesia, while it showed gradual decrease from second hour to 7th day after anesthesia. 8) By oxygen inhalation the Opsonin rate showed a slight decrease temporally, but it recovered at the end of inhalation. 9) The influence of various general anesthesia upon the Opsonin rate in blood was independend of the blood coagulating time and the number of the platelets. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 69 2 1957 全身麻醉の生体防禦機能に及ぼす影響に関する研究第1編各種全身麻醉の動脈血及び静脈血血液凝固時間ならびに血小板数に及ぼす影響に関する研究 349 357 EN Shinobu Yumiyama 1) The basal narcosis using Ravona, Amithal and 2% Pantopon scopolamine made no variation of blood coagulating time and platelets number. 2) During the intravenous anesthesia with Penthothal sodium the blood coagulating time was delayed temporally directly after the anesthesia. Concurring with it the number of platelets decreased but soon recovered. At this time the decrease of platelets was striking especially in the venous blood. 3) During the endotracheal anesthesia with cyclopropaine the blood coagulating time was reduced one hour after the anesthesia temporally. It was then delayed with the progress of anesthesia. The blood platelets increased and decreased according with it. They were striking in the venous blood. 4) During the endotracheal anesthesia with ether the blood coagulating time showed a distinct reduce with the progress of anesthesia and even after the end of the anesthesia its recovery was delayed. The blood platelets distinctly increased and its recovery was also distinctly delayed. These variations were characteristically more striking in the arterial blood. 5) By the anesthesia using cyclopropaine and nitrous oxide together the variation of blood coagulating time and platelets number were alike to that during the anesthesia with cyclopropaine, but they were very variable after the anesthesia. 6) The platelets variation in all kind of anesthesia was parallel with that of blood coagulating. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 12-2 1959 アスピリンアレルギーに関する実験的研究第3編アスピリン蛋白に依る局所性皮膚過敏性に就いて 8419 8428 EN Susumu Kimura There are already many reports in various allergic reactions by aspirin, but most of them deal with the reactions induced by aspirin alone or a physical compound of aspirin and serum protein, as the antigens. For this reason they failed to demonstrate the antigen-antibody reaction in vitro although they recognized the establishment of active allergy. In addition, there are few reports in which conjugates of aspirin to serum proteins are used as antigen, but there seems to be no report on any experimental localized passive allergy. In the present experiment, the author selectively sensitized rabbits and guinea-pigs using conjugates of aspirin to proteins or aspirin solutions as the antigens, and studied the localized skin allergy in these test animals. The results are as follows. 1. In the rabbits and guinea-pigs sensitized with aspirin-protein, active and passive Arthus' reactions have been recognized. 2. In the above-mentioned active and passive Arthus' reactions, a cross reaction has been recognized to occur by heterogous antigen just as in the case of the precipitation reaction. 3. In the sensitization of guinea-pigs using aspirin alone, it has been recognized that there occurs the active Arthus' reaction by aspirin-proteins and by aspirin. 4. In the active Arthus' reaction induced by the aspirin-protein, there can be recognized some reaction differences between the sensitizing antigen and the cross-antigen. The grade of these reactions is more marked in the former. 5. In the active Arthus' reaction induced in the guinea-pigs sensitized with aspirin alone, no reaction differences such as mentiones above can be recognized. 6. In the passive Arthus' reaction there can't be recognized some reaction differences between the corresponding antigen and the chemical cross antigen. 7. The animals sensitized either actively or passively with aspirin have been found to respond to sodium salicylate. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 12-1 1959 皮膚疾患に於ける血中並びに皮膚中電解質に関する研究第2編実験的家兎皮膚炎に於ける血中並びに皮膚中電解質量の変動に就いて 7897 7908 EN Kakuso Kunihara Following the course of the dermatitis caused with croton oil in the back of rabbits, sodium, potassium and calcium in sera and skin were determined spectrophotometrically. 1. The quantity of sodium in sera showed a slight decrease in the initial stage of the dermatitis, showed increase in the severe stage and moreover increased in tho retrogressive stage and returned to the normal value in the recovering stage. In skin, increasing from the initial stage, showed marked increase in the severe stage and then decreased rapidly and showed nearly normal value in its retrogressive stage. 2. The quantity of potassium in sera increased slightly and showed remarkable increase in the severe stage, but, decreasing rapidly, showed almost normal value in retrogressive stage. The variation of potassium in skin showed the same tendency as sodium but, in the initial stage, increase of potassium was more remarkable than sodium. 3. Calcium in sera showed aslight increase in the initial stage, showed the lowest value in the rotrogressive stage and then gradually increased but returning to the normal value was delayed. In skin, showed a slight increase in the initial stage, showed the lowest value in the severe stage but thereafter was elevated gradlualy. 4. The value of Na/K in sera showed the lowest in the severe stage and in skin the lowest was showed in the initial stage and the both declined early and were eleavtated later. 5. K/Ca in sera showed the highest value in the retrogressive stage and, in skin, the highest was showed in the severe stage and the both were elevated in early period and dropped in the latter half. 6. Almost similar tendency of the variation of electrolytes was showed in the sera and the skin, but in latter more remarkable. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 味覚の臨床的研究第2編血液含有物質，胃液酸度及び唾液pHと味覚との関係に就て 7409 7424 EN Kimiaki Honda The corrlation among the cotained substance in blood, the acidity of fastric fluid the pH of saliva and the taste was observe on the attacked paiod and convalescen pariod of disease. And the resulte were as follows. 1. Observing on the correlation between the dosis of serum sodium and the throshold value of taste, the threshold value of sweed taste showed the high value in many cases with the low value of serum sodium at the attacked period of disease and it showed the declining tendency with the increase of serum sodium by the recovery of disease. No fixed correlation between the dosis of serum sodium a d the salt, sour or bitter taste was observed. 2. The correlation between the dosis of serum chloride and the threshold value of saste was similar to that on the occasion of serum sodium. 3. As for the correlation between the dosis of serum potassium and the threshold value of taste, many cases with the high value of serum potassium at the attacked period of disease showed the high value of serum potassium at the attacked period of disease showed the high threshold value of sweet and bitter taste and they showed the returning tendency to normal value at the convalescent perio with normal value of serum potassioum. No fixed correlation between the dosis of serum potassium and the threshold value of salt or sour taste was observed. 4. As for the correlation bet ween the dosis of serum iron and the threshold value of taste, the threshold value of bitter taste showed the tendency taking the high value on the low value of serum iron. The tendency was especially remerkable in the patients with anemia. No fixed relation between the dosis of serum iron and the threshold value of sweet, salt or sour taste was not observed. 5. As for the correlation between the dosis of serum copper and the threshold value of taste, the theshold value of sweet and sour taste showed the tendency to declins, as the dosis of serum copper became decreased at the attaked period of disease in many cases. And they showed the tendency to decline with the decrease of serum copper at the convalescent period, but no fixed correlation between the desis of rerum copper and the threshold value of salt or bitter taste. 6. The correlation between the increase or decrease of blood sugar at the hunger time and the rise of decline of the threshold value of sweet or sour taste was observed on various diseases, other than diabetes, In diabetes, the threshold value of sweet taste at the hunger time showeed the high valun, but no fixed correlation between the blood sugar at the hunger time and the threshold value of salt or bitter taste was observed. 7. No fixed correlation among the dosis of serum protein, the acidity of gastric fluid, the pH of saliva the threshold value of taste was observed. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 皮膚疾患に於ける血中並びに皮膚中電解質に関する研究第1編各種皮膚疾患に於ける血中電解質量の変動に就いて 7291 7304 EN Kakuzo Kunihara In order to explain the physiological conditions in skin diseases by searching the tendency of metabolism of electrolytes, the contents of sodium, potassium, calcium and chlor in serum were measured in 122 cases of various skin diseases. (1) Comparatively many cases of the numerous skin diseases showed the variation of the contents of electrolytes, but it was not always remarkable and constantly regular. (2) In bullous diseases, acquired pigmentary disturbances, diffuse scleroderma and erythroderma, varied the most remarkably, and especially in the 2 former cases was pointed out the evident tendency. (3) In eczema, dermatitis, inflammatory keratosis, erythematodes and cutaneous tuberculosis were often found abnormal values, but very irregular and complicated. (4) In urticaria, drug eruption and erythema exsudativum multiforme showed slight changes. (5) The auther conceived that the metabolism of electrolytes in vivo was so influenced by the several organs, especially endocrine organs, and by the autonomous nervous system, that such irregularity of this variation was showed. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 Bromsulphalein試験に関する知見補遺第2編 B. S. Pの胆汁中排泄に及ぼす血液pH，有機物，無機塩類の影響 7267 7278 EN Seishin Iwami 1. The higher the pH of serum was, the more was the serum bromsulphalein value and the bromsulphalein value excreted into bile was decreased. 2. There was no special correlation between the serum pH and colloidal reaction. 3. The remarkable changes of bromsulphalein dosis excreting into bile and of serum bromsulphalein value were not observed on the increase of urea dosis in blood, in the canine with the fistula of gallbladder. 4. The serum bromsulphalein value became a little high on the increase of lactic acid dosis in blood, in the same canine, and the bromsulphalein dosis excreted into bile was considerably decreased. 5. The serum bromsulphalein value became a little high on the increase of uric acid dosis in blood, in the same canine, and the bromsulphalein dosis excreted into bile was considerably decreased. 6. The special changes of serum bromsulphalein value and bromsulphalein dosis excreating into bile were not observed on the increase of sodium in blood, in the same canine. 7. The serum bromsulphalein value became decreased on the increase of potassium in blood, in the same canine, and the bromsulphalein dosis excreted into bile was increased. 8. The serum bromsulphalein value became a little high on the increase of calcium in blood, in the same canine, but the remarkable change of bromsulphalein excreted into bile was not observed. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 Bromsulphalein試験に関する知見補遺第1編 Bromsulphaleinと血清蛋白質との結合に関する研究 7253 7265 EN Seishin Iwami 1. The absorption maxima at 593, 590 and 588 mμ were observed on the coloration by the N/100 caustic soda solution at the pH 7.5 after the addition of bromsuplhalein into human serum albumin, blood plasma and serum. 2. The combined rate of bromsulphalein with albumin was conversely increased with the decrease of bromsulphalein density on the combination of human serum protein and bromsulphalein by the utilization of paper electrophoresis and the rate of wondering pigment was gradually decreased, but the rate of residual scanty bromsulphalein combined with globulin or attached in the filter paper was almost fixed. 3. Observing the influence to the combination of serum albumin and bromsulphasein on the change of various buffer used for the paper electrophoresis, the combination was obstructed in order of veronal soda, hydrochloric acid, primary potassium phosphate, secondary natrium phosphate, borax, boric acid and veronal soda buffer. 4. Observing the influence to the combination of serum albumin and bromsulphalein on the change of pH after the addtion of buffer into normal human and dilutied serum with distilled water, the combination of them were rapidly dcreased around the pH 8-9 and it became almost zero around the pH 12. 5. In the previous experiment, the combination was more decreased with the addition of urea, lactic acid or uric acid in each than without their addition, but it showed the fixed combination by the kind of buffer on the addition of urea. the extremely scanty decline of combination on the addition of lactic acid and the considerably remarkable decline of combination. 6. Sodium chloride gave no influence of the combination of albumin and bromsulphalein in the serum of same pH, potassium chloride acted to increase the combination, and calcium chloride acted to bromsulphalein itself and acted to control the coloration by caustic soda. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 Bilirubinと蛋白との結合に関する研究第2編 Birirubinとglobinとの結合に就いて 7163 7169 EN Masanosuke Ohtsuki In order to study the combination of bilirubin with globin and also the combination of bilirubin with globin+albumin, the author estimated the absorption curves by means of paper electrophoresis using veronal buffer solution at pH 8.5 as the electrolyte and by paper chromatography at the first dimension using a 2% cane sugar solution as a developer, as well as by D-K type Beckman's autospectrophotometer; and obtained the follo wing results. 1. Sodium bilirubinate that is a direct bilirubin and dibasic acid bilirubin, an indirect bilirubin, both haye the capacity to combine with with globin. 2. Such combinations of bilirubins and globin do not control the mode of bilirubin to the diazo reaction. 3. When sodium bilirubinate is dissolved in the mixed solution of globin and albumin, it will combine with either one, namely, with globin and with albumin. Moreover, in this instance the mode of the proteins just as in the mode of sodium bilirubinate, shows a direct response to the diazo reaction but never an indirec response. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 11-1 1959 Bilirubinと蛋白との結合に関する研究第1編 Bilirubinとalbumin及びglobulinとの結合について 7155 7161 EN Masanosuke Ohtsuki With the purpose to study the combinations of bilirubin with albumin and with globulin using a veromal buffer solution (pH 8.5) as the elctrolyte the author estimated the absorption curve by means of paper electrophoresis and by paper chromatography at the first dimesnion using 2% cane sugar solution as the developer and with D-K type Beckman's autospectro-photometer; and obtained the following results. 1. Sodium bilirubinate that is a direct bilirubin and dibasic acid bilirubin which is an indirect bilirubin both combine with albumin. 2. The mode how bilirubin combines with albumin can not be the controlling factor of the direct or the indirect bilirubin reaction. 3. When desalting is done on the combined solution of sodium bilirubinate and albumin, a mixed solution of bilirubin and albumin is obtained, but no bilirubin can be isolated. 4. It is quite difficult to combine blirubin with globulin. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 10-2 1959 Urobilin体の定量に関する研究第1篇 Urobilin体定量法の検討 6953 6965 EN Haruo Watanabe The Terwen's reduction method and the methods reported by Heilmeyer, Krebs and Watson were fully observed. And the results were as follows. 1. Urobilin was completely reduced to urobilinogen within 2 hours by the Terwen's method, and the highest uroblinogen value was observed on the mixed rate of the materials and the reducing agents in 40:5:5. The materials should be stored, under the isolation of air and light, at the low temperature as possible and there was a limitation for the preserved time. 2. The optimal range of pH for the extraction of urobilinogen into the solvont. 3. It was demonstrated that petroleum ether was the best solvent for the extraction of urobilinogen since the comparative studies on the various solvent, especially between ether and petroleum ether. 4. The composition of regent needed the concentration of 0.3% P-dimethylaminobenzaldehyde on the use of 60% hydrochloric acid and of 1% P-dimethylaminobenzaldehyde on the use of 50% hydrochloric acid. 5. The shaking process for one minute was needed till the addition of sodium acetate after the addition of regents, urobilinogen in the solvent was extracted into hydrochloric acid in the regent and the reaction was performed in the water layer by the above treatment. The progress of aldehyde reaction on this occasion was considerably quantitative. 6. The significance of adding sodium acetate was to shift to nearly the pH 4 after the terminal reaction and was to remove the coloration by indole body, with bringing the coloration of urobilinogen up to the highest and keeping the stability of it. 7. The influence of indole, indican, P.A.S. and sulfonamid derivatives shown the similar reaction in the qualitative test could be eliminated on the quantitive method. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 10-1 1959 試験内胆汁色素生成に関する研究第2編 Oxyhaemoglobin及びCarboxyhaemoglobinの1-Ascorbin酸と分子酸素による分解過程に関する分光化学的研究 6587 6594 EN Shoji Tarumi By studying spectrochemically the decomposition processes of carohemoglobin, oxyhemoglobin, methemoglobin, and carboxyemoglobin in the presence of 1-ascorbic acid and molecular oxygens the author obtained the following results. 1. In the case of carbohemoglobin besides tha already-known absorption peaks of 556 mμ and 430 mμ it shows another absoorption peak at 760 mμ, and its EmM is 1.4-2.0. 2. The absorption curve of the 670 mμ and 630 mμ substances at the time of deoxydation by sodium dithionite and causatic soda show the absorption peak at 618 mμ and 760 mμ. 3. The decomposition of caroxyhemoglobin takes similar decomposition processes as taken by oxyhemoglobin and methemoglobin, producing the 670 mμ and 630 mμ substances. In other words, it seems that caroxyhemoglobin is reduced to hemoglobin by liberating carbon monoxide and then it enters into decomposition processes, and its decomposition is extremely slow as compared with that of oxyhemoglobin and methemoglobin. 4. CO-choleglobin is not an intermediate product of carboxyhemoglobin decomposition but it is produced at the time when choleglobin comes in contact with carbon monoxide. 5. CO-choleglobin is split into carbon monoxide and choleglobin in the presence of 1-ascorbic acid and molecular oxygens. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 10-1 1959 試験内胆汁色素生成に関する研究第1編 Methaemoglobinのl-Ascorbin酸と分子酸素による分解過程に関する分光化学的研究 6577 6585 EN Shoji Tarumi By amking 1-ascorbic acid and molecular oxygens act upon methemoglobin the author studied the decomposition processes spectrochemically and obtained the following results. 1. In preparing methemoglobin from oxyhemoglobin with sodium nitrite no spectrochemical effect of sodium nitrite can be recognized on methemoglobin. 2. Methemoglobin is reduced to oxyhemoglobin in the presence of 1-ascorbic acid and molecular oxygen and thereafter it shows a similar decomposition processes as observable in the case of oxyhemoglobin. 3. This reaction is affected by molecular oxygens namely, when the quantity of molecular oxygen is large, the speed of the reaction is greater but the production of 670 mμ substance is inhibited; whereas when the quantity of molecualr oxygen is small, the absorption index of 670 mμ and 630 mμ substances is greater. 4. The 670 mμ and 630 mμ substances that are intermediate substances of this reaction are reversible to one another depending upon the quantity of moecular oxygen present. 5. This reaction is affected by the ion concentration; namely, between pH 6.8 and pH 7.7 at pH 7.2 the 670 mμ and 630 mμ substances are produced in the greatest amounts. 6. This reaction is affected by 1-ascorbic acid; namely, with the increase in the amount of ascorbia acid the speed of reaction is accelerated. 7. This reaction is affected also by temperature; namely, with a rise in temperature the reaction is accelerated whereas with a fall in temperature the reaction is retarded. 8. At an eary stage of this reaction dipyrromethen serlies substances are produced by the decomposition of the 670 mμ and 630 mμ substances. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 10-1 1959 ツベルクリン反応に関する研究第1編消化並びに化学的変性に依るツベルクリン抗原性の変化について 6485 6490 EN Shiro Mishima By making protein degenerant and protein digestive enzyme act on old tuberculin the author performed the tuberculin reaction, and obtained the following results. 1. In loading a strong potassium iodide solution to old tuberculin the tuberculin reaction grows weaker along with an increase in the concentration of potassium iodide. This suggests that free tyrodine radical in the old tuberculin protein is involved in the tuberculin reaction. 2. In loading sodium nitrite to old tuberculin the tuberculin reaction likewise grows weaker with increase in the concentration of sodium nitrite 3. In the addition of formalin to old tuberculin the tuberculin raction is weakened along with increaes in the concentration of formalin solution. 4. When crystalline trypsine is made to act on old tuberculin, the tuberculin reaction is weakened along with lapse in the digestion time. 5. When pepsin is made to act on old tuberculin, the tuberculin reaction is likewise weakened along with lapse in the digestion time. 6. From these results it is deduced that the tuberculin reastion is mainly controlled by tuberculin protein. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 9-2 1959 犬肝臓の細胞内顆粒からのin vitro Histamine遊離に関する研究 6173 6186 EN Kei Jinzenji In vitro histamine release from intracellular large granules of the dog liver was determined and compared with that from the chopped tissue. The histamine release from the large granules depended on the concentration, duration of action, temperature and pH of histamine liberators. Histamine was released by all the liberators tested even at 0°C less than at 37°C. The action of basic liberators such as sinomenine increased with increasing alkalinity of medium. Dibucaine hydrochloride, decylamine hydrochloride, HgCl(2), quinine hydrochloride and tutocaine hydrochloride at pH 7-8 and 6mM concentration released over 50 per cent of histamine from the large granules. Under the same conditions ethylmorphine hydrochloride, toluidine blue, procaine hydrochloride, saponin (0.1 per cent), tropacocaine hydrochloride, sodium cholate, Compound 48/80 (0.1 per cent), sinomenine hydrochloride, Tween 20 (0.2 per cent), cocaine hydrochloride and xylocaine hydrochloride revealed the histamine liberating action in the descending order named. Histamine release from the granules by these substances was larger in variable degrees as compared with that from the chopped tissue. Sodium salicylate inhibited the release from the granules and chopped liver by other substances, while diphenhydamine and guaiazulen did not reveal such an inhibitory action but rather accelerated the release though slightly. The histamine release from the chopped tissue by sinomenine and decylamine was inhibited by uranil nitrate, but it was not the same from the large granules. The lack of oxygen accelerated the histamine release by decylamine from both the granules and chopped tissues, but did not reveal any significant effect on the action of other liberators. In in vitro anaphylaxis histamine release occurred in the chopped tissue but not in the granules. As far as basic liberators are concerned, there seemed to be some correlation between the histamine release ability and the heparin combining power. However, the surface activity or the hemolytic power of liberators and the histamine release ability was not to be necessarily in parallel with each other in degree. Interpreting the intracellular large granules as the granules of mast cells and in the light of the above findings the author discussed the respective mode of action of the liberators on mast cells and on their granules. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 9-2 1959 沃度酸曹達による網膜変性症の組織化学的研究第3報脂質並びにミトコンドリアの変化に就て 5921 5928 EN Yasuta Koyama The author studied on the changes of the lipid and mitochondria in experimental degeneration of the retina of the rabbit caused by intravenous injection of 5% solution of sodium iodate. Lipids in the pigment epithelinm slightly decrease with the exception of those in the lipochrin. No remakable change was observed in the lipids of the outer segments and ellipsoid of vesual cells. Most of the mitochondria in pigment epithelial cells disappeared as the cells degenerate, but some of them remained unchanged even in rather advanced stages of degeneration. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 9-1 1959 沃度酸曹達による網膜変性症の組織化学的研究第2報クロームケトエノール物質並びにD. N. A. の変化に就て 5695 5704 EN Yasuta Koyama The author studied on the early histochemical changes of the peculiar retinal degeneration of the rabbit treated with intravenous injection of 5% solution of sodium iodate as the preceding report. The Cr-K E S (chrmo-keto-enol substance of HAMAZAKI), chief constituents of which are nucleotides of D N A, in the pigment epithelium and outer segments of rods begins to diminish gradually after 12 hrs. D N A decreases as the pigment epithelium and visual cells degenerate. In general, the Cr-K E S diminishes and disappears earlier than the Hg-K E S in the pigment epithelium and in visual cells. The authers that sodium iodate acts rather indirectly on the metabolism of the D N A and related substances in the outer layers of the retina. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 8-2 1959 痙攣時における犬大脳皮質の各種電解質の変動に関する実験的研究 5363 5374 EN Hiroshi Yorimae Changes of the electrolyte in the motor area of the cerebral cortex were experimentally studied in dogs during convulsions which were caused by the intravenous injection of metrazol. The results were as follows: 1) Chloride content increases at the preconvulsive stadium and also at the acme and reaches to its maximum at the repeated convulsive stadium. At the postconvulsive stadium it decreases nearly to the normal, but still remains a little higher than the normal 2) Sodium content decreases at the preconvulsive stadium and recovers gradually to the normal as the time elapses. 3) Potassium content shows no distinguished change. 4) Calcium content reaches the highest at the preconvulsive stadium and then decreases gradually to the normal as the time elapses. 5) Magnesium content at the preconvulsive stadium is the same as the normal and increases rapidly at the acme and at the repeated convulsive stadium but finally decreases below the normal. 6) From these facts stated above, it is considered that the changes of the electrolyte in the cerebral tissue during the convulsion are caused by the secondary changes due to convulsion. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 8-1 1959 再生不良性貧血における副腎皮質機能に関する研究第3編患者の副腎皮質機能 4613 4627 EN Jiro Kurozumi With a view to clarify the cause of hypoplastic anemia the author studied the adrenal cortical function in patients with this disease, and obtained the following results. 1. The basic metabolic rates show not any definite inclinations. 2. In the insulin tolerance test, the insulin index has been found to have declined. 3. As for electrolytes in serum, chlorine is decreased but phosphorus is increased to a high degree, and ratio Na/K is slightly high while calcium, sodium and potassium show no definite inclinations. 4. Robinson-Power-Kepler water test is almost positive. 5. On measuring 17-KS and free chemocortioids (Ch. C.) excreted in the urine, the amount of 17-KS is decreased conspicuously but Ch. C. is increased. 6. In the gel-ACTH loading test the adrenal cortical reserve function has been found to have declined highly. 7. From these facts stated above, it is possible to assume that there is a decline in the adrenal cortical function in this disease. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 6-2 1959 呼吸酵素系の組織化学的並に生化学的研究第1編呼吸酵素系によるテトラゾリウム塩の還元機構について 3601 3608 EN Hiroaki Okazaki With the purpose to establish the integrating methods for the selective histochemical demonstration and colorimetric estimation of each enzyme and enzyme system belonging to the terminal respiratory enzyme system, an alytical study on the reduction of neotetrazolium chloride by tissue homogenates was carried out in the present report using various substrates and inhibitory agents, and the steps with which the reaction of neotetrazolium chloride reduction conjugates were determined as follows. 1. The reaction of neotetrazolium reduction by tissue homogenetes using sodium succinate as substrate is not the succinic dehydrogenase reaction but the succinoxidase system reaction; and the reaction takes place cojugating about 50 per cent in the step of the succinic dehydrogenase system, of these about 15 per cent conjugates in the step prior to the antimycin A sensitive step and 35 per cent in the step itself; and about 50 per cent in the step of cytochrome c oxidase. 2. In the case using p-phenylenediamine as substrate the reaction of neotetrazolium reduction is the reaction due to the activity of cytochrome c-cytochrome oxidase system, and when p-phenylenediamine is used with the sufficient amount of cytochrome c, the reaction appears to be dependent on cytochrome c oxidase activity. Neotetrazolium reduction in all these reactions takes place conjugating in the step of cytochrome c oxidase. 3. In the case where DPN and substrates taking DPN as a coenzyme are used, the reaction of neotetrazolium reduction is mainly the reaction conjugating at the step below antimycin A sensitive step in the DPNH-cytochrome c reductase system. 4. Endogenous dehydrogenase reactions are the sum total reactions conjugating at the steps prior to the antimycin A sensitive step in the terminal electron transport system and with other various reduction systems which are not inhibited by antimycin A. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 6-2 1959 肉芽嚢及び人工結合織嚢の吸収能に関する研究 3403 3418 EN Motoyuki Shigemasa As a step to investigate the pathological physiology of joints, the author attemped to study the absorption of sodium salicylate and carbon black in granuloma pouch (Selye) of rats and artificial connective tissue pouch (Terazawa) of rabbits and obtained the following results: 1. The absorption of sodium salicylate in the granuloma pouch of rats and in the artificial connective tissue pouch of rabbits is very rapid. 2. The absorption of 20%-polyvinyl pyrrolidon-sodium salicylate is slower than in the former. 3. In the pouch of the hydrocortisone-protected rat (Selye), the proliferation of the granuloma tissue and carbon absorption in the granuloma pouch are inhibited. 4. When the daily injection of 100mg/kg sodium salicylate is administered intraperitoneally to rats, the proliferation of granuloma tissue and the carbon absorption are slightly inhibited. 5. After the injection of hydrocortisone into the rat granuloma pouch, necrosis can be observed on the surface of the pouch wall, proving that there is a little carbon absorption. 6. Although the carbon absorption in the pouch having remarkable fibrosis is retarded, it can be accelerated by the scraping or washing of the pouch wall. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 6-2 1959 低体温麻酔および低温下手術における電解質の変動に関する実験的研究第2編低温下手術における電解質の変動に関する実験的研究 3357 3369 EN Masatoshi Mizushima Effects of operation on seruml evel and urinary excretion of sodium, chloride and potassium, and water and electrolytes coutent in tissue were studied on dogs in both hypothermic and normothermic conditions. Following results were obtained. 1) Hyponatremia and hypochloremia, decrease of urinary excretion of both electrolytes, increase of potassium-sodium ratio and increase of urinary excretion of potassium in accordance with decrease of urine volume were found during postoperative period in dogs who underwent operation under normothermia. However, increase of urine volume occurred on about 5th or 6th postoperative day resulting in increase of urinary excretion of sodium and chloride, and decrease of potassium excretion. Increase of water, sodium and chloride content of the tissue which had operative intervention, were found on the 4th postoperative day, though decrease of potassium was noted. 2) Decrease of serum sodium and chloride, and increase of potassium were noticed in hypothermic group, though these changes were not so significant as in nomothermic group. Urinary excretion of sodium and chloride increased in accordance with increase of urine volume and chloride-sodium ratio were diminished. However, changes after 5th to 6th postoperative day were same in both groups. There were found no significant changes in water and electrolytes content of tissue, which had operative intervention, on the 4th postoperative day in hypothermic group. Increase of water, sodium, and chloride in tissue were slight and no decrease of potassium was found. It was anticipated based on these findings that excretion of ADH and function of adrenal gland were suppressed resulting in increase of urinary excretion of sodium and chloride with less retention of water, sodium and chloride in tissue during postoperative period in operation performed under hypothermia. In addition to, decrease of potassium content in tissue was not found, suggesting of less consumption of body protein and suppression of catabolism during postoperaetive period. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 6-2 1959 低体温麻酔および低温下手術における電解質の変動に関する実験的研究第1編低体温麻酔における電解質の変動に関する実験的研究 3345 3356 EN Masatoshi Mizushima Changes in serum and urine electrolytes, sodium and potassium in erythrocytes, water and electrolytes contents in tissue were studied in dogs under hypothermia. Following results were obtained. 1) Changes in serum sodium and calcium are not found, though potassium decreases and chloride increases in hypothermia. 2) No marked changes in sodium and potassium in erythrocytes are found. 3) Urinary excretion of sodium and chloride decrease, while potassium increases. No changes in excretion of calcium are noted. Total amount of electrolytes excreted in urine decrease because urine volume is markedly diminished in hypothermia below 28°C. 4) No significant changes in water content in tissue are found. There is found no changes in sodium of tissue. Chloride in tissue decreases, while potassium in liver increases though no changes are found in other organs. Calcium in tissue remains constant. Decrease of serum potassium and increase of chloride are remarkable changes in electrolytes in hypothermia. It is anticipated that deposition of potassium develops in liver because of decrease of glucose utilization due to low metabolism and decreased consumption of glycogen with resulting glycogen formation from excessive glucose which carries potassium from the extracellular fluid into the cells. Increase of serum chloride is supposed to be due to mobilzation of intracellular chloride into blood and decrease of excretion in urine. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 6-2 1959 ラットにおけるHistamineの尿中排泄に関する研究第2報 Histamineの尿中排泄に対する諸種Histamine遊離物質及びHistamine遊離に影響を及ぼす物質の作用 3289 3299 EN Kazuji Kondo With the administration of histamine there occurs a transient increase of the urinary excretion of free histamine in the female rat being loaded with water and whose urine being collected every 30 minutes. In the previous paper (Folia pharmacol. japon. 54, 1221, 1958) it was reported that such an increase is dependent upon the basal rate (a) of the urinary excretion of histamine and the amount (b) of histamine excessively excreted after intraperitoneal injection of a limited amount of histamine and that there is a relationship of b/a=k (k stands for constant pertaining dose). On the basis of these findings and taking the increase in the urinary excretion of histamine as an index, the author compared the potency and time course of in vivo histamine release as demonstrated by sinomenine hydrochloride, Compound 48/80, quinine hydrochoride, sodium cholate, dextran, egg white, decylamine and Tween 20, administered intraperitoneally. Judging from the relationship between the degree of the increase in the urinary excretion of histamine and the amount of the substances being administered, the histamine-releasing ability of these substances was found to be in the order of; Compound 48/80>decylamine>sinomenine>sodium cholate>quinine, Tween 20>dextran and egg white. Of them, Compound 48/80 and sinomenine showed the increase in the urinary histamine exactly identical with that observed after histamine administration, taking the shortest time course; while in the cases of quinine, sodium cholate, dextran, and egg white the increase was somewhat slower; and by decylamine and Tween 20 it developed most slowly yet most persistently. In the rats previously treated with guaiazulene, cortisone, aminopyrine or cinchophen sodium, increases in the urinary excretion of histamine by all the releasers mentioned above were similarly inhibited. Since mechanisms of histamine release action are not the same by different releasers, it seems that the action of these inhibitors is of such a nature as to be manifested at a common stage involved in different patterns of the mechanism of histamine release. Such an action of cortisone was more marked in a relatively small dose rather than in a large dose, suggesting presence of an adequate amount of this steroid to be used for the manifestation of this action. The histamine-releasing ability of egg white was inhibited in the alloxan-diabetic rat, while on the contrary, it was greatly accelerated in the insulin-treated rat. Such an effect of alloxan was completely antagonized by an adequate amount of insulin. However, in the rat with glycosuria induced by glucose administration, sensitivity to egg white was not altered. Succinic acid or oxaloacetic acid given with the purpose to inhibit the formation of keton bodies, could not eliminate the above mentioned alloxan effect. Both alloxan and insulin did not in any way modify the histamine-releasing effect of sinomenine. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 4-1 1959 本態性萎黄貧血の下垂体副腎皮質系に関する研究第3編 Screening Testによる物質代謝について 1237 1251 EN Shomi Yamamoto With a view to clarify the cause of idiopathic hypochromic anemia the author studied the function of the adrenocortical system, this time, by observing the changes in the tissue metabolism through the screening test. 1. The basal metabolic rate showed a slight increase and it was mcstly increased in males while it was mostly at the normal level in females. 2. As for serum inorganic substances chlorine was decreased but sodium and phosphate were increased; and the ratio Na/K was high, showing irregular tendency in the function of the pituitary adrenocortical system involving the mineral metabolism. 3. By Robinson-Power-Kepler water test 29.4per cent showed positive rate, indicating a slight fall in the function of the pituitary adrenocortical system involved in hydroelectrolytic metabolism. 4. In the insulin tolerance test the insulin tolerance index and a decline in insulin resista nce and recovery of hypoglycemia were found to have decreased slightly. However, by treatment the insulin resistance improved but the recovery of hypoglycemia rather became worse. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 4-1 1959 Pentothal・Sodium静脈麻酔時間と自律神経機能に関する実験的研究第3編自律神経剤によるChE活性値および循環動態の変動がPentothal・Sodium体内分布に及ぼす影響に関する実験的研究 1165 1180 EN Yoshio Matsuda 1) The increased cholinesterase activity coursed by administration of Acetylcholine, pilocarpine, Imidalin and chloromazine reduce the concentration of pentothel in the Brain and Serum. 2) The decreased cholinesterase actriity coursed by administration of physostigmin, prostigmin and DFP increase the concentration of pentothal in the Brain and Serum. 3) The mutation of distribution of pentothal in the Body coursed by administration of Ephedrin, Epinephrin and Norepinephrin should be attributable to both increased Blood pressure and reduced cholinesterase activity. 4) The mutation of anaesthetic duration and the distribution of pentothal in the body changed by administration of various drugs of autenomic nervous system should be attributable to both cholinesterase activity and Blood pressure influenced by these drugs. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 71 4-1 1959 Pentothal・Sodium静脈麻酔時間と自律神経機能に関する実験的研究第2編自律神経毒剤加のPentothal・Sodium体内分布に及ぼす影響に関する実験的研究 1151 1164 EN Yoshio Matsuda Following the estimation of anaesthetic duration, I investigated the distribution of pento thal in the body (Brain, Blood, Kidney, Leber, Muscle. Fat), taking advantage of Beck-mantype Spectroptomoter. 1) Epinephrin, Norepinephrin, Ephedrin coursed the increased coucentration of pentothal in the Brain, Kidney, Muscle and Fat, and coursed the decreased concentration in the blood. 2) The prolonged duration of anaestetic coursed by administration of prostigmin, physostigmin, D. P. E. are shown to be correlateed with an increased rate of penetration of anaesthetic in the blood and brain. 3) Pyruvate also prolonged the duration of anaesthetic and this prolongation is reduced by addited administration of acetylcholin. These above results show that the inhibition of cholinacetylase by pyruvate coursed the increased permeability to pentothal and that addition of acetylcholin reverse the effect. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 101 5-6 1989 HIV-1エンベロープ蛋白質の大腸菌での発現と精製 659 672 EN Bo Zhang To purify the HIV-1 envelope protein with antigenic reactivity, the Pvu II-Bgl II fragment of the HIV-1 env gene, from the Pvu II site to the second Bgl II site, encoding the carboxyl terminal 180 amino acids of the viral surface protein (SU, gp120) was molecularly cloned in Escherichia coli strain HB101 using protein A expression-shuttle vector pRIT5. The pRIT5 contains the protein A gene, encoding the secretion signal and IgG binding domain of protein A with the upstream promoter and the downstream multicloning sites, as well as the two replication sites for Escherichia coli and Staphylococcus aureus. A fused protein with the molecular weight of about 55 kilodaltons was produced, which showed the same reactivity as the native protein A against rabbit serum IgG on Western blotting analysis. Most of the fused protein in the periplasmic space was degraded, while the complete fused protein inside the cells was recovered as an insoluble protein. The fused protein was solubilized with sodium dodecylsulfate (SDS), partially purified by IgG sepharose affinity chromatography, and completely purified by SDS-polyacrylamide gel electrophoresis. The quantity of the expressed fused protein was estimated about 1% of the total proteins. The purified fused protein contained 516 amino acids with Mr54, 976, consisting of 305 amino acids of the IgG binding domain of protein A, 5 amino acids derived from polylinker, a carboxyl terminal 180 amino acids of the HIV-1 envelope surface protein gp120, and 26 amino acids derived from the pUC19 sequence. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 102 11-12 1990 単純ヘルペスウイルス1型潜伏感染の研究における弱毒株の有用性 1253 1259 EN Yujiro Arao The usefulness of avirulent strains of herpes simplex virus type 1 (HSV-1) in investigations of latency was examined. Trigeminal ganglia (TG) latently infected with HSV-1 were obtained more efficiently from mice inoculated with an avirulent strain, SKO-1B, than from those inoculated with a virulent strain, F. Moreover, utilization of two avirulent strains, SKO-1B and -GCr Miyama, allowed us to compare the rate of reactivatable latency (RL) in these strains. Cultivation of explants of TG recovered from mice inoculated with the same dose of either infectious virus revealed a significant difference in the rate of RL between the two strains. Both strains were revealed to be neither hypersensitive nor resistant to acyclovir, indicating that the difference in the rate of RL between the two strains was not due to any deficient activity of viral ribonucleotide reductase and/or thymidine kinase. The recovery rate of the -GCr virus from explants of TG latently infected with this virus was greatly enhanced by treatment of the explants with sodium n-butyrate during in vitro cultivation. As such, at least one of the mechanisms responsible for causing a very low rate of RL in TG infected with -GCr Miyama may be reduced expression of immediate-early genes of this virus in latently infected cells in vitro. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 102 7-8 1990 著明な低カリウム血性ミオパチーをきたした原発性アルドステロン症の一例 843 849 EN Yuko Ohtani Tadashi Yoshino Tadaatsu Akagi Takashi Kitagawa We report a rare case of primary aldosteronism with severe hypokalemic myopathy in a 59-year-old female who had suffered from hypertension since 1979. Because of severe muscle weakness, especially of extremities, she became unable to rise from her bed and was admitted to the Kitagawa hospital on July 10, 1989. Symptoms such as tetany, polyuria, and diarrhea were not observed. Her consciousness was clear. The blood pressure was in the range of 152 mmHg systolic and 104 mmHg diastolic. Systolic heart murmur (Levine II), cardiac arrythmia, and slight edema on the legs were noticed. She did not have any sign of muscular atrophy. Serum aldosteron level was 220 pg/ml, and plasma renin concentration was less than 0.5 ng/ml/h. The serum level of potassium was 1.7 mEq/l, sodium 150 mEq/l, GOT 47 IU/l, LDH 507 IU/l, CPK 850 IU/l (MM type 99%), and aldolase 9.2 mIU/ml. Computed tomography (CT) after intravenous contrast injection revealed a round low density mass (1.6 cm in diameter) in her left adrenal gland. CT also revealed the gallbladder adenomyomatosis. Her left adrenal gland and gallbladder were surgically removed on July 26, 1989. After the operation, the blood pressure and laboratory data including serum potassium returned to normal, and she became able to pursue her ordinary life. Endoscopy revealed a gastric polyp which was hyperplastic. Histopathologically the removed adrenal gland showed an adrenocortical adenoma consisted of clear cells and nodular hyperplasia of the glomerular zone. 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 12 1953 放射能泉に関する研究（XXXIII）放射能泉入浴の血中鉱質に及ぼす影響 34 41 EN Sakue Onda 10.18926/40431 In order to study the effect of radioactive thermal bath on mineral substances in blood, the author bathed rabbits in Misasa Hot Spring (Yamada-Yu) once daily for 5 minutes dnring the successive 3 weeks, and determined potassium, calcium, sodium, chloride and protein levels in serum before and after the bath every week. The radon content of Yamada-Yu was 300-600×10(-10) curies per liter, its water temperature being 42-44°C. at that time. In the early stage of serial baths calcium decreased, and potassium, sodium, chloride and protein content increased. But in the later stage a reversed tendency was recognized. It was concluded that radioactive thermal baths of Misasa had no peculiar effect on mineral substances in rabbit's serum, compared with the effects of the other kinds of thermal baths 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 11 1953 溫泉の老化に関する研究（第1報）三朝溫泉の湧出後のラドン含量の変化 1 13 EN Takeshi Sugihara 10.18926/40412 The author determined the radon content of the thermal waters of Misasa, Tottori Prefecture Japan, at various times after flowing out, and obtained the following results: 1) The velocity of variation is expressed by a simple equation with respect to the radon content. At thermal temperaturee of 41.5-41.8°C and room temperature of 24.3-28.0°C, a following exrerimental equation was obtained, u:;ing a beaker with a inner diameter of about 17 cm and a height of 27 cm, log a = -0,0048t + 2.59 a: radon content in Mache unit. t : time in minute. 2) A close relation was observed between the room temperature and the rate of variation of radon, and also between the thermal temperature and the rate of variation of radon, that is, the rate of variaton of radon was proportional to the room temperature and the thermal temperature respectivly, as it is clearly seen from the distribution coefficient of radon. 3) There exists a c1ose relation between the velocity of variation of radon and the surface area being in contact with air, that is, the velocity was generally proportional to the square root of the surface area, or to the radius of the surface. 4) The velocity of variation of radon content was roughly inversely proportional to relative humidity of air. 5) Betweeen the range of sodium chloride concentrations from 0.1 to 40.1 g. per liter no difference was proved concerning the velocities of variation in radon content. 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 9 1953 胸やけに就て 54 63 EN Masazumi Sotozono 10.18926/40386 402 consecutive patients with various disorders of digestive tract were investigated concerning heartburn and 184 cases (45%) proved to have heartburn. No significant relationship was established between the heartburn and gastric acidity or the kinds of digestive diseases. Heartburn was not only seen in hyperacidity but also often seen in hyp-and anacidity or normacidity. No significant correlation was observed between the heartburn and the rate of emptying the gastric content. Flow back of bile in stomach juice was relatively often seen in cholecystopathic patients with heartburn, but no such relationship was proved in other kinds of patients. Experimentally beartburn was evoked in the patients with heartburn by administration of 30 cc of 1% sodium bicarbonate solution in the stomach or ten cc in oesophagus, but not by the same quantity of hydrochloric acid (N/20), lactic acid (N/20), 10% syrup, 50% polytamine solution, physiological saline, or 5% bile dilution. A habit of eating fast was observed relatively often in heartburn patient. Profession of the most of the above examined patients was farmer and they noted some causative relation to large intake of food rich in carbohydrate (sweetpotato, rice-cake, etc. ). The most frequent time for heartburn to beginn was one to three hours after a meal. 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 12 1953 温泉の老化に関する研究（第3報）　溫泉水中の珪酸塩の溶存狀態 6 10 EN Takeshi Sugihara 10.18926/40353 When silicate content is determined by the colorimetric method, the silicate content in the mineral waters and sodium silcate solution decreases in addition of ammonium hydroxide and sodium carbonate respectively, and moreover after addition of aluminium ion in the samples, when aluminium hydroxide is precipitated from ammonium hydroxide solution, total silicates in various forms are coprecipitated with it. The silicate content in the mineral waters that are kept in concentrations of 0.1～0.4 normality of sodium hydroxide showed special variations with times. 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 13 1953 人工放射性同位元素による温泉作用の研究 (3)　浴水中の硫酸イオンの体内進入に及ぼす連続浴，火傷治癒経過，色素塗布並びに硫酸カルシウム水溶液の影響 18 23 EN Takeo Yokota Takashi Ashizawa 10.18926/40345 By using labelled sodium sulfate or calcium sulfate (sulfate containing S(35)) the transition of sulfate ion into the body across the skin was investigated soon after taking a bath in sodium sulfate or calcium sulfate solution under varing conditions. Mice were used for experiments. The percutaneous absorption of sulfate ion proved to decrease gradually in the course of serial baths in sodium sulfate solntion and was accelerated by making a fresh burn on the skin of the bathed animals, but it showed a tendency to decrease as the wound became more and more healed. The application of basic dye to the skin (e.g. 1% methylenblue or 1% fuchsin solution) increased the transition of sulfate ion into the test animal. Little difference was proved between the bath in sodium sulfate and calcium sulfate solution with respect to the penetration of sulfate ion. Passage of sulfate ion from the bath water through the skin was a little promoted by the use of sodium sulfate solution than by the use of calcium sulfate solution. 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 13 1953 鳥取県三朝温泉に於ける二，三の放射性元素の存在について 5 8 EN Mitsuo Sato 10.18926/40342 The author tried to measure RaB, RaC and ThB in the hot spring waters using G-M counter and proved the existence of RaB and ThB in Misasa Hot Springs. To 100 liters of spring water ahout 20 gms of ferric chloride and then sodium hydroxide were added. Precipitate was dissolved in hydrochloric acid and after adding bismuth and lead, the solution was saturated with hydrogen sulfide. The precipitate of sulfides was filtered and ignited to ashes after drying. The β-activity of the ashes was measured by a mica-window type G-M counter with the recording circuits of scale of 16. Natural background was 50±5 connts per minutes. Six spring waters of Misasa, namely "Spring of Branch Laboratory", "Nakayu", "Gunze" "O-T-R", "Jwayu", and "Tsukiminoyu" were investigated. RaB was detected in all samples. ThB was found only in "Gunze" and "O-T-R". ThB content of "Gunze" was estimated, to be roughly 1×10(-12) Curie units per liter. 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 14 1954 鳥取県下の温泉のラドン含有量に就て 1 14 EN Yoshio Oshima Naoharu Yamada Masaaki Mifune 10.18926/40324 1. Radon content of 166 thermal waters in IWAI, TOTTORI YOSHIOKA, HAMAMURA, TOGO, SEKIGANE, MISASA, and KAlKE Hot Springs was measured by I. M. Fontactoscope in the years 1950-1951. Of which 66 samples showed a radon content over 30×10(-10) curie units per liter. Namely, 50 springs in Misasa, 6 in Sekigane, 9 in Hamamura, and one in Togo belonged to the radioactive spring in the definition by Ministry of Social Welfare. The highest Radon content (1150×10(10) curie units per liter) was recorded in Hisuino-Yu in Misasa, where five springs had a radon content over 360×10(-10) curie units per liter. 2. No marked difference was proved between the results obtained this time and the data in the former reports concerning the radon content of these thermal springs. 3. Radon content proved higher in the springs which issue from granite than in the springs of other districts. No definite relation was proved between the radon content and water temperature. The radon content was generally high in simple thermals or in weak sodium chloride springs, low in sulfated springs and in saline springs which had a comparatively high sulfate content. 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 22 1958 温泉医学領域に於ける濾紙分析法の研究　（1）濾紙電気泳動法並びに濾紙chromatographyによる血清鉄の研究 37 54 EN Maruo Ishibashi 10.18926/40265 1. The author showed by means of paper-electrophoresis using o-nitrosoresorcinmonomethylether (hereafter N. R. M. E.), a colourdeveloping reagent, that iron in serum combines with β-fraction of serum protein. 2. The author proposed a new method for the determination of serum iron by means of paper-chromatography. Procedure is as follows. In a centriguged tube, 0.5 ml. of clear serum is taken, acidified with 0.15 ml. of 20% HNO(3), and allowed to stand for 20 minutes. After addition of 0.2 ml. of saturated solution of CH(3)COONa and 1 ml. of acetic acid-sodium acetate buffer solution (pH: 3.8), the mixture is kept for at least 10 min. and filtered through Toyo-filter-paper No.5 A. The clear filtrate is taken in a stoppered test tube (the inside diameter: 1.2 cm.; height: 15 cm.). To this filtrate are added 0.2 ml. of 20% Hydroxylamine hydrochloride and 0.25 ml. of saturated aqueous solution of N. R. M. E. After being allowed to stand for at least 20 min., the mixture is shaken with 5 ml. of carbon tebrachloride. Then the excess of N. R. M. E. is removed. The supernatant aqueous solution (0.2 ml.) is used for a determination sample. The sample is placed on the paper (Toyo,filter-paper No.51 or 50) in a thin line at a distance of 10 cm. from the edge, and the spots are developed with 90 % ethylalcohol. After development for 4-6 hours, a green-coloured linear spot appsars at a distance of 12-16 cm. from the origin-point. Spot intensity at 670 mμ is measured with Natsume's densitometer, and from this, concentration of Fe can be estimat.ed on the standard graph (Fig. 5). 3, Effects of radioactive thermal spring bathing and internal use of vitriol water upon serum iron levels were studied with the result.s as follows. The above-mentioned author's method of determining the iron level in a small amount of serum is very convenient in investigating iron metabolism in the field of balneotherapy. The author examined changes of iron levels in serum after radioactive thermal bathing as well as after internal use of acid vitriol water. a) The iron levels in serum were maasurecl by the author's method before and 5, 30 and 60 minutes after the radioactive thermal bath (Rn-content: 10-30 Mache, 42-3°C., for 10 min,). The iron levels in the serum of healthy subjects showed no significant change after the thermal bathing. b) Yanahara Mineral Water (an acid vitriol water, pH; 2.2) contains 0.045 gm. of Cl(-), 12.35 gm. of SO(4)(--), 2.5 gm. of Fe(++)+Fe(+++), 0.0938 gm. of Al(+++) and 0.0001 gm. of Cobalt in one liter. i) Thirty ml. of Yanahara, water diluted with plain water to 200 ml. (Fe(++) content: ca. 75 mg.) and administered to health fasting subjects orally. As a control matter, 0.65 gm. of Glukon-F powder (Fe(++) content.: ca. 75 mg.) was then given. Blood samples were taken from the cubital vein before and 1, 3 and 6 hours after the intake of the dtriol water. Iron contents of the serum samples were measured by the author's paper chromatographic method. The results were shown in the table and the figure. The iron contents of the serum after the intake of the mineral water were higher than in the case of the control. ii) Two ml. of Yanahara vitriol water was administered to rabbits with the aid of a stomach tube, and serum iron levels wera determined before and 1, 3, 4 and 6 hours after the administration. An amount of 0.05 gm. of Glukon-F powder was dissolved in 5 ml. of plain water, and this solution was given to rabbits as a control. After the internal use of vitriol water, iron levels in serum rose and remained unexpactedly high for about 1-4 hours, in a manner that did not. correspond to the amount of iron in the given mineral water. So that it was suspected that the iron originally contained in the living subjects had been mobilized by the intake of vitriol water. Iron levels in serum after administration of a solution of Glukon-F powder were lower than in the case of the mineral water. 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 23 1958 鳥取県東郷松崎温泉に関する研究 1 22 EN Shunji Umemoto Mitsuru Harada Shigeru Okabe Junichiro Miyakoshi Masanobu Sakanoue Masaya Tanaka Mifune Masaaki 10.18926/40259 1. Layers containing thermal water in this district are thin, and lie at different depths (about 35, 55, and 60 meters) from the ground surface. There are evidences to show that these layers are intimately connected with one another. 2. The authors may suppose the existence of a structurally weak zone, along the line from Matsuzaki to Asozu, within which the issuing spots of thermal springs are located. 3. The head water levels of the thermal springs in this district are closely related with that of Lake Togo. Keeping pace with the variations of the water levels of Lake Togo and of artesian wells in its vicinity, the rate of flow of thermal springs vary; and the correlation between these variations is apparent. 4. The pumping suction of thermal water at one spring affects the flow of water at other springs within distances of 150 to 200 meters therefrom, though the direct sources of thermal water supply for the latter springs may be different from that of the former. 5. The spring water in this district is considered to be a mixture of hot water, containing sodium, calcium, chloride, and sulfate ions, and cold water, containing bicarbonate ion. The diversity of chemical constitutions of different spring waters is explained as due to the difference in proportion in which the hot and cold waters are mixed. 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 41 1972 慢性関節リウマチの金療法に関する研究 III 原子吸光分光分析法による検討 35 48 EN Kazue Takahashi 10.18926/40152 The gold concentration in serum, synovial fluid and urine has been determined using a HITACHI MODEL 207 atomic absorption spectrophotometer. Flow rate of acetylene was setted at 2.0l/min. and lamp current was setted at 8mA. The samples were pretreated by the determination of gold. Serum was diluted with deionized water by two to ten-fold. To O.2ml. of synoival fluid 50units/ml of hyaluronidase was added and then incubated for 20 minutes at 37℃. This sample was then treated the procedure for serum given previously. Urine was treated using the modification method of Christions procedure. The recovery rates with gold sodium thiomalate in serum, synovial fluid and urine were 99.9%, 102.5% and 93.6% respectively. When gold sodium thiomalate was injected in rabbits, the maximum serum gold level was attained 1 hour after injection and then gradually decreased at about 50% of the maximum level 24 hours after injection. In the observations of changes in the gold levels in the serum of patients with rheumatoid arthritis after injection of gold sodium thiomalate and gold thioglucose, its peak level after gold thioglucose injection tended to be slightly lower as compared with the same amount of gold sodium thiomalate injection. In those patients who had been given 300 mg. or more of gold salt-preparation, one additional injection of 25 mg. of gold salt could attain the gold level of around 200μg/100ml, in serum a week later, and this level was very nearly the same with that after 3-4 days of one additional injection of 1Omg. of gold salt. The amount of gold excreted in the urine varied from patient to patient, and the mean excretion rates of gold sodium thiomalate and gold thioglucose in the urine within 24 hours after intramuscular administralion were 9.8% and 6.5% respectively. The simple, rapid and accurate method for analysis in biological specimens using atomic absorption spectrophotometer would offer one of the excellent measures in the clinical management of patients with rheumatoid arthritis. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 60 51 2004 Reactivity of TEMPO anion as a nucleophile and its applications for selective transformations of haloalkanes or acyl halides to aldehydes 11969 11975 EN Tsutomu Inokuchi Hiroyuki Kawafuchi Sodium 2,2,6,6-tetramethylpiperidine-N-oxide (TEMPO−Na+), generated by reduction of TEMPO· with sodium naphthalenide in THF, reacted with alkyl halides or acyl halides to produce O-alkylated or acylated TEMPOs, which were in turn oxidized with mCPBA or reduced with DIBAL-H to afford the corresponding aldehydes, thus accomplishing a new protocol for the halides-carbonyls conversion. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 136 9 1988 Studies on the acid-base properties of the AIBr3-NaBr melts 2606 2608 EN Hidetaka Hayashi N Hayashi Z Takehara A Katagiri The acid-base properties of the ZnBr[2]-NaBr melts at 623 K were investigated on the basis of the electromotive force measurements of a zinc-zinc concentration cell. The following two chemical equilibria were postulated to describe the acid-base character of the melts ZBnr[2]+Br[-]=ZnBr[3][-] K[1] ZnBr[3][-]+Br[-] =ZnBr[4][2+] K[2] The equilibrium constants K[1] and K[2], were determined to be 5.0×10, and 1.0×10[2], respectively, at 623 K No potential conflict of interest relevant to this article was reported.

Pergamon-Elsevier Science Ltd. Acta Medica Okayama 0016-7037 68 24 2004 A theoretical interpretation of the chemical shift of Si-29 NMR peaks in alkali borosilicate glasses 5103 5111 EN Tokuro Nanba Mitsunori Nishimura Yoshinari Miura In Si-29-NMR, it has so far been accepted that the chemical shifts of Q(n) Species (SiO4 units containing n bridging oxygens) were equivalent between alkali borosilicate, and boron-free alkali silicate classes. In the sodium borosilicate glasses with low sodium content. however. a contradiction was confirmed in the estimation of alkali distributions B-11 NMR suggested that Na ions were entirely distributed to berate groups to form BO4 units, whereas a -90 ppm component Was also observed in Si-29-NMR spectra, which has been attributed to Q(3) species associated with a nonbridging oxygen (NBO). Then. cluster molecular orbital calculations were performed to interpret the -90 ppm component in the borosilicate, glasses. It Was found that a silicon atom which had two tetrahedral borons (B4) as its second nearest neighbors was similar in atomic charge and Si2p energy to the Q(3) species in boron-free alkali silicates. Unequal distribution of electrons in Si-O-B4 bridging bonds was also found. where much electrons Were localized oil the Si-O bonds. It was finally concluded that the Si-O-B4 bridges with narrow bond angle were responsible for the -90 ppm Si-29 component in the borosilicate glasses. There still remained another interpretation: the Q(3) species were actually present in the glasses. and NBOs in the Q(3) species were derived from the tricluster groups. such as (O3Si)O(BO3)(2). In the classes With low sodium content. however. it was concluded that the tricluster groups were not so abundant to contribute to the -90 ppm component. No potential conflict of interest relevant to this article was reported.

Acta Medica Okayama 45 1 2007 Sorption Characteristics of Honeycomb-Type Sorption Element Composed of Organic Sorbent 183 191 EN Hideo Inaba Takahisa Kida Akihiko Horibe Makoto Kaneda This study deals with the sorption characteristics of a honeycomb-type sorption element composed of a new organic sorbent that was composed of the cross-linked polymer of sodium acrylate. Transient experiments in which moist air was passed into the honeycomb-type sorption element were conducted under various conditions of air velocity, temperature, relative humidity and honeycomb length. As a result, the effective mass transfer coefficient of the organic sorbent adsorbing the water vapor was non-dimensionalized as a function of Reynolds number, modified Stefan number and non-dimensional honeycomb length. No potential conflict of interest relevant to this article was reported.

Springer Berlin / Heidelberg Acta Medica Okayama 1618-2642 387 6 2007 S,S,S-Tris(2-ethylhexyl) phosphorotrithioate as an effective solvent mediator for a mexiletine-sensitive membrane electrode 2057 2064 EN Takashi Katsu Yumi Tsunamoto Nobumitsu Hanioka Keiko Komagoe Kazufumi Masuda Shizuo Narimatsu S,S,S-Tris(2-ethylhexyl) phosphorotrithioate proved to be an effective solvent mediator for constructing a mexiletine-sensitive membrane electrode in combination with an ion-exchanger, sodium tetrakis[3,5-bis(2-methoxyhexafluoro-2-propyl)phenyl]borate. Among a series of phosphorus compounds containing phosphoryl (P=O) groups, this solvent mediator showed the highest sensitivity to mexiletine in phosphate-buffered physiological saline containing 0.15 mol L-1 NaCl and 0.01 mol L-1 NaH2PO4/Na2HPO4 (pH 7.4), giving a detection limit of 2 x 10(-6) mol L-1 with a slope of 58.8 mV decade(-1). This is the best reported detection limit of any mexiletine-sensitive electrode developed to date. Owing to its high selectivity toward inorganic cations, the electrode was used to determine the level of mexiletine in saliva, the monitoring of which is quite effective for controlling the dose of this drug noninvasively. The mexiletine concentrations determined with the mexiletine-sensitive electrode compared favorably with those determined by high-performance liquid chromatography. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 24 1 1970 A morphologic study of protein ingestion by Ehrlich ascites tumor cells 27 48 EN Koiti Sogabe Article 10.18926/AMO/32798 For the purpose to obtain the information of the mechanism of protein uptake by the tumor cells, some cytochemical and electron microscopic observations were carried out on Ehrlich ascites tumor cells incubated with horseradish peroxidase (basic hemoprotein, molecular weight approximately 40,000) in vitro. In the earlier periods of the incubation peroxidase was found to be adsorbed on some area of surface of the tumor cells forming a thin protein layer, where an active pseudopodia formation was observed. With the lapse of time, the protein was taken in the deep cytoplasm by the infoldings of the cell membrane and accumulated in the cytoplasmic vesicles having limiting membrane. Concerning the accumulation of the protein into the vesicles, small tubular structures in the cytoplasm connecting the cell surface and the vesicles, were considered to participate in the intracellular transportation of peroxidase taken up. In cold environment (2°C), the formation of pseudopodia and deep inward infoldings of the cell membrane was inhibited and simultaneously the uptake of peroxidase stopped. Iodoacetate and sodium fluoride also effected to suppress the pseudopodia and infoldings formation moderetely, as well as uptake of peroxidase, though they did not stop completely. These facts have indicated that horseradish peroxdase is taken up by Ehrlich ascites tuimor cells through pinocytosis which involves energy-requiring process dependent upon glycolytic metabolism of the tumor cells. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 24 2 1970 Binding of photosensitizing dyes with some biopolymers 185 204 EN Shigeo Yasui Article 10.18926/AMO/32769 To obtain some information of the biological action of Kankohso 101dinicotinate and Kankohso 301-nicotinate, observations were made on the binding mode of these substances with protein, chondroitin sulfate and nucleic acids and the following results were obtained; 1. Kankohso 10 I-dinicotinate binds reversively with bovine serum albumin or serum r-globulin, resulting in metachromasia. By binding with proteins the absorption maximum of the dye shifts toward the long wave length side and the absorbance decreased distinctly. The data show that there are more than one kind of binding sites and the binding with bovine serum albumin is weak in acidic solution and strong in alkaline solution. 2. Kankohso 10 I-dinicotinate produces strong metachromasia with sodium chondroitin sulfate and the color of the solution changes from violet blue to reddish violet. The absorption maximum at 592 mp. decreases without shifting its wave length ,and the shoulder appears at 555 mp. be. comes distinct peak. The strongest metachromatical changes occurs at the concentration of the chondroitinsulfate whose sulfonate radicals is equal to the molecules of Kankohso 10 I-dinicotinate. 3. Kankohso IOI-dinicotinate produces metachromasia with nucleic acid, where absorption spectrum is shifted toward long wave length and absorbance is decreased at a certain concentration. 4. Kankohso 301.nicotinate binds weakly with bovine serum albumin, the binding of which is reversible and the maximum binding number is 1.1 per molecule of albumin. Metachromasia cannot be produced by binding. Kankohso 30I.nicotinate does not bind with bovine serum γ-globulin. This compund does not produce metachromasia with sodium chondroitin sulfate but produces weak metachromasia with nucleic acid, indicating some affinity to nucleic acid. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 46 3 1992 Concentration and uptake of taurine in umbilical blood platelets. 169 174 EN Hirofumi Kanemori Kohei Ejiri Shuichiro Akahori Takafumi Kubo Kaoru Sekiba Article 10.18926/AMO/32671 The concentration and uptake of taurine in the umbilical and adult blood platelets were studied. Taurine was the most abundant free amino acid in both umbilical and adult blood platelets. The taurine concentration in umbilical blood platelets (2.30 pmoles/10(4) cells) was significantly lower than that of adult blood platelets (3.27 pmoles/10(4) cells) in contrast to the reverse relationship in taurine concentrations in umbilical and adult blood plasma. No other amino acid showed such significant difference in the concentrations between umbilical and adult blood platelets. Taurine uptake into umbilical blood platelets was temperature sensitive and sodium-dependent in a manner similar to that of adult blood platelets. The uptake conformed well to Hanes-plot. The Vmax of the uptake into adult blood platelets was about 3.6 times higher than that of umbilical blood platelets, but no significant difference was seen in the Km value between the two groups. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 22 3 1968 A method of column chromatographic isolation of major phospholipid components on Escherichia coli 147 152 EN Tieko Yosioka Kazuya Akatsuka Akira Yamagami Yasuhiro Kanemasa Article 10.18926/AMO/32583 For the column chromatographic isolation of individual phospholipids from the total phospholipid mixture, silicic acid, DEAE cellulose, alumina and others, have been used as adsorbent. However, it must be emphasized that silicic acid (1, 2, 3, 4) is the most useful adsorbent for the separation of the total phospholipid mixture from each other in reasonable purity. VAN DEENEN reported that pure phosphatidyl glycerol was obtained from the lipid fraction of spinach leaves after repeated chromatography on silicic acid column (5). The phospholipid extracted from Escherichia coli B consists of abundant phosphatidyl ethanolamine (70-80 %), cardiolipin, phosphatidyl glycerol and other minor components as described in the previous paper (6). The high percentage content of phosphatidyl ethanolamine renders it difficult to separate the phospholipids by the column chromatography. Therefore, repeated chromatographies on the silicic acid column treated with sodium bicarbonate (7) and normal silicic acid column were employed for the isolation of the major components from the total phospholipid of E. coli B. Stepwise elution (4) was carried out with chloroform containing increasing proportions of methanol, and the eluent was divided into several fractions according to experience with thin-layer chromatography. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 22 6 1968 A light and electron microscopic study of the distribution of gold sodium thiomalate in the rheumatoid synovial membranes 293 317 EN Hajime Inoue Article 10.18926/AMO/32571 The synovial membranes from 16 rheumatoid patients treated with intramuscular injections of gold sodium thiomalate were observed by light and electron microscopy with special reference to the distribution of gold particles in the tissue. 1) Light microscopic study revealed that the gold demonstrated as cytoplasmic granules by OKAMOTO'S histochemical method were contained in the synovial lining cells and in the macrophages around lymph-follicles and blood vessels in the subsynovial layer. In the well-developed villi on the surface of rheumatoid synovial membrane, large macrophages with gold granules infiltrated into the lymphoid cell accumulation of small lymph-follicles. 2) The deposition of gold in the synovial tissue increased with the increase of the doses of gold administered. 3) Electron microscopic observation indicated that gold particles are contained in the numerous lysosomes in the Type A and intermediate lining cells. The macrophages around lymph-follicles and blood vessels also possessed a large amount of gold particles gathered in the lysosomes of these cells. 4) Macrophages containing gold particles in their long cytoplasmic extensions were found often in a close contact with plasma cells of various differentiation stages. A direct cytoplasmic connection was observed between the two kinds of cells but an artifact could not be excluded. 5) The effect of gold salt in the treatment of RA was discussed from the immunological view point. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 23 1 1969 Effects of distention of the small intestine on the movements of the gallbladder and the sphincter of Oddi 21 26 EN Sosogu Nakayama Article 10.18926/AMO/32559 1. Dogs anesthetized with pentobarbital sodium were mainly used and effects of the distention of the small intestine on the movements of the gall bladder and the sphincter of Oddi were investigated. 2. The distention of the small intestine (jejunum or ileum) inhibited the rhythmic contraction of the gall bladder and duodenal movements, and relaxed the tone of the sphincter of Oddi, resulting in an increase of the outflow of fluid through the orifice of the common bile duct. 3. After cutting the bilateral thoracic splanchnic nerves together with extirpation of the bilateral upper lumbar sympathetic trunks, the inhibitory response on the movements of the gall bladder and the tone of the sphincter of Oddi was completely abolished. The vagus nerve did not take part in the reflex response described above. The transection of the spinal cord at the level between Thl and Th2 produced no change in the reflex responses. 4. Fwm the results described above it may be supposed that effects of the distention of the small intestine on the movements of the gall bladder and the sphincter of Oddi are produced via the thoracic and lumbar splanchnic nerves through the reflex center which is located in the spinal cord. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 23 3 1969 The competitive effect of adenosine-5'-triphosphate against the stimulating and inhibiting actions of 2,4-dinitrophenol on the mitochondrial respiration 227 235 EN O. Hatase G. Yamamoto T. Oda Article 10.18926/AMO/32549 Effect of ATP and substrates on 2,4-dinitrophenol-induced adenosine triphcsphatase (E. C. 3.6. 1. 4.) activity and respiration of isolated rat liver mitochondria has been investigated. 1. The oxidation of sodium succinate inhibited the action of 2, 4-DNP on the induction of adenosine triphosphatase activity in the mitochondria. 2. A moderately large amount of sodium succinate restored the suppressed mitochondrial respiration due to 2, 4-DNP. 3. Adenosine-5'-triphosphate (ATP) restored quantitatively the released and inhibited mitochondrial respiration due to 2,4-DNP, and its prior addition prevented also quantitatively the action of 2,4-DNP on the mitochondrial oxygen up-take. These ATP effects were oligomycin sensitive, and they were considered to manifest their actions through the phosphorylation system. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 23 5 1969 Ultrastructure and biochemical function of the mitochondria in respiratory-deficient mutant yeast induced by 4-nitroquinoline nitrogen oxide 343 356 EN Kimikazu Koshiba Takuzo Oda Hiromichi Tsukamoto Nobuyuki Goto Article 10.18926/AMO/32546 1. A respiratory-deficient mutant strain of yeast was obtained from wild strain of Saccharomyces servisiae by treatment with 4-nitroquinoline N-oxide. Ultrastructure and function of the wild or mutant strains and the mitochondrial fractions isolated from these strains were examined by biochemical and electron microscopic analyses. 2. The frequency of the respiratory-deficient mutant strain in yeast induced with 10-6M 4-nitroquinoline N-oxide was about 40 %. 3. Respiratory-deficient mutant strain is incapable of reducing 2, 3, 5-triphenyltetrazolium chloride salt and to grow on lactate medium. In addition to this, the mutant has been found to have lost its ability to take up oxygen in sodium succinate and pyruvate. 4. 4.Nitroquinoline N-oxide in the concentration that induces a mutant of yeast cells or its kin inhibits the oxygen uptake in normal strain. 5. The normal strain of yeast is characterized by difference spectrum corresponding to cytochromes a+as, band c+Cll respectively, whereas, the mutant strain containes almost no cytochromes a+ as, band C1 but contains normal or increased amount of cytochrome c. 6. Mitochondrial fraction isolated from mutant strain has largely lost its ability to oxidize succinate. On the other hand, NADH-, lactate-and cytochrome c-oxidase activities are reduced by about 1/17, 1/7 and 1/8 of that of normal strain, respectively. 7. Succinate dehydrogenase activity of mutant strain is almost zero. Moreover, this activity is not affected on the addition of phenazine methosulfate. NADH dehydrogenase activity of mutant stran is about 1/2 of normal strain. 8. The variations in mitochondrial structure of normal and mutant strain in the stationary phase have been followed with the aid of electron microscopy. In contrast to the normal strain, the mutant strain revealed distinct morphological changes in mitochondria, especially, the lack of cristae in its interior. The results have been interpreted to indicate that the mutant induced by 4.nitroquinoline N.oxide has a character of cyto. plasmic mutant. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 23 4 1969 ATP synthesis of submitochondrial particles driven by proton gradient 291 302 EN Osamu Hatase Article 10.18926/AMO/32520 l) The submitochondrial particle system can synthesize ATP in the early phase (220 seconds after the accition of ADP) in the presence of sodium succinate and Pi, in spite of the absence of the hexokinase-glucose system, and this phosphorylation is inhibited by oligomycin. 2) The submitochondrial particle system can synthesize ATP by the base-acid transition (proton pulse) only in the presence of ADP and Pi, in spite of the absence of oxidizing substrates and the hexokinase-glucose system, and this phosphorylation is dependent on the span of pH change, and is inhibited by oligomycin and 2, 4-dinitrophenol. 3) The role of the proton vector in the oxidative phosphorylation and the proton ejection was discussed from the stand point of a new hypothesis. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 18 5 1964 Effect of sodium oleate on the metabolism and size of rat liver mitochondria I. Uncoupling, shrinkage and their reversal action by bovine serum albumin 247 259 EN Goki Yamamoto Article 10.18926/AMO/32472 Effects of sodium oleate and bovine serum albumin (BSA) on rat liver mitochondrial function and structure were studied by measuring oxygen uptake, 90° light-scattering, adenosine triphosphatase activity and pyridine nucleotides fluorescence. 1. The low concentration of oleate induced the uncoupling of oxidative phosphorylation and the scattering change of mitochondria. This action of oleate differed from that of oleate at a high concentration which induces the high amplitude swelling with respect to its physiological and biochemical properties. The degrees of reversal swelling (shrinkage) and of oxygen uptake induced by oleate in the presence of Pi and succinate were altered proportionately to the concentration of oleate, and the concentration of oleate to the shrinkage coincided with that of the maximal respiratory release. 2. Antimycin A or 2, 4- dinitrophenol prevented the oleate-induced mitochondrial shrinkage, but the treatment of these agents after prior incubation with Pi and succinate allowed the shrinkage, though the degree was small in its extent compared with that in the absence of inhibitors. On the other hand, oligomycin did not affect the shrinkage with oleate. 3. BSA protected the mitochondrial phosphorylation from the uncoupling action of oleate without showing any effect of its own. A complete reversal could readily be demonstrated by a sufficient amount of BSA from the uncoupling, structural changes, and oxidation of intramitochondrial pyridine nucleotides induced by oleate in a low concentration. 4. The oleate-stimulated latent ATPase activity was proportional to the oleate-induced shrinkage of mitochondria with respect to the concentration of oleate. The latent ATPase was abolished also by the addition of a sufficient amount of BSA. 5. The action of oleate on the phosphorylation sequence of mitochondria was discussed on the basis of the present findings. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 18 4 1964 Effect of cystathionase on isovalthine 239 240 EN Toshihiko Ubuka Katsumi Horiuchi Takehira Shimomura Tsukasa Azumi Brief Note 10.18926/AMO/32451 In the course of studies on the cleavage reaction of S-(isopropylcarboxymethyl) glutathione (GSIV) into isovalthine in kidney homogenate or glutathionase preparation, it has sometimes been observed that the amount of isovalthine formed is far less than that of GSIV decomposed¹. Furthermore, when such reaction mixture is analyzed on an automatic amino acid analyzer, prominent peak corresponding to the reasonable amount of S-(isopropy1carboxymethyl)cysteinylglycine which is an expected intermediate of the GSIV cleavage reaction cannot be found up to 400 effluent ml. Though several reasons may be considered for the explanation of the above curious phenomenon, the effect of cystathionase on isovalthine is at first examined here. But the result was negative. L- and L-Alloisovalthineused as substrate were prepared by the method of OHMORI². Homoserine and purified cystathionase in ammonium sulfate solution prepared according to the method of GREENBERGB³ were kindly furnished by Prof. M. Suda of Osaka University. Incubation mixture contains 0.1 ml of enzyme solution, 1.0 ml of 0.2 M borate buffer (pH 8.0) containing 2×10-³M cysteine, 0.lml of 0.1 M substrate, and 0.8ml of deionized water containing 5×10-4M EDTA. The mixture was shaken at 37°C for 30 minutes in the air. The reaction was terminated by adding 2ml of 10% trichloroacetic acid and the α-keto acids formed were determined by the method of FRIEDEMANN and HAUGEN4 with a following modification: toluene extract was washed once with 8 ml of 10% sodium sulfate. The results obtained are summarized in Table l. When the reaction mixtures are analyzed before or after incubation on an automatic amino acid analyzer, the amount of L- or L-Alloisovalthine is found to be unchanged. Furthermore, as indicated in Table 1, L-isovalthine showed no inhibitory effect on the homoserine cleavage by cystathionase. Since amino acid oxidases have already been reported to have no effect on isovalthine³, the curious phenomenon above cited may have to be explained by other reaction mechanism such as transpeptipation reaction. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 16 6 1962 Mitochondrial Swelling and Uncoupling Activity of Long-Chain Fatty Acids 317 331 EN Kozo Utsumi Sachiko Ohara Goki Yamamoto Kozo Inaba Hiroyuki Urakami Michio Yamamoto Article 10.18926/AMO/32359 The effect of various fatty acids on the swelling-contraction and oxidative phosphorylation of mitochondria from rat liver and Ehrlich ascites tumor cell have been studied and the results are as follows: 1. The swelling of rat liver mitochondria is induced by fatty acid. The extent of this uncoupling action is in the descending order of myristate, laurate, parlmitate, stearate and behenate in saturated fatty acid and linoleate, linoleneate, richinoleate and oleate in the unsaturated fatty acid. This swelling action is stronger with unsaturated fatty acids than that of saturated ones and cis form is stronger than trans form. 2. The uncoupling oxidative phosphorylation of rat liver mitochondria is also observed with these fatty acids and the activities are proportional to the degree of the swelling action. 3. The degree of swelling of rat liver mitochondria is proportional to the concentration of oleate and is inhibited by anaerobiosis and respiratory inhibitor except amytal. 4. The mitochondria swollen by fatty acid can be recontracted reversibly by ATP, Mg++ and bovine serum albumin. 5. The swelling action of sodium oleate is the strongest on mitochondria from rat liver, followed by those from the liver of Ehrlich ascites tumor bearing mouse, Ehrlich ascites tumor cells and solid Ehrlich tumor cells. 6. Sodium oleate inhibits the incorporation of 32p into ATP, ADP, GTP and UDPG in mitochondria. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 54 2 2000 Possible postsynaptic action of aminoglycosides in the frog rectus abdominis. 49 56 EN Yusuf Karatas Yusuf Ergun Cemil Gocmen Ata Secilmis Ergin Singirik Atilla Dikmen Firuz Bayasal Article 10.18926/AMO/32289 The present study was undertaken to investigate the postsynaptic effects of aminoglycosides on contractions evoked by acetylcholine (ACh), KCl, electrical field stimulation (EFS) and Na(+)- and Ca(2+)-free Ringer solution with 0.2 mM Na2 EDTA (NaFCaFR) in the isolated frog rectus abdominis. Neomycin inhibited contraction elicited by ACh, NaFCaFR, and EFS at the higher frequencies (8 and 10 Hz) but not those elicited by KCl and EFS at the lower frequencies (2, 3 and 5 Hz). D-tubocurarine inhibited ACh-induced contractions in a concentration-dependent manner. In addition, drug reduced EFS-evoked contractions to a limited extent. Lower concentrations (10(-5), 5 x 10(-5), 10(-4), 2 x 10(-4) and 3 x 10(-4) M) but not higher concentrations (4 x 10(-4) and 5 x 10(-4) M) of methoxyverapamil exhibited a concentration-dependent inhibitory action on NaFCaFR-induced contractions. Similar inhibitions of the same type of contraction were displayed by aminoglycosides (neomycin, streptomycin, netilmycin, gentamycin and amikacin). These results suggest that in addition to their antagonistic action on nicotinic receptors in the frog rectus abdominis, aminoglycosides may exert stabilizing effects on some functional components contributing to contractions at the membrane. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 54 2 2000 Purification, identification and phosphorylation of annexin I from rat liver mitochondria. 57 65 EN Kenji Yoshii Katsuyoshi Sugimoto Yuji Tai Ryoji Konishi Masaaki Tokuda Article 10.18926/AMO/32286 Annexin was purified from rat liver mitochondria to an apparent homogeneity with a molecular weight of 35 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified mitochondrial annexin (AXmito) was identified as annexin I by an immunoblot analysis using anti-annexin I antibody. The inhibitory effect of AXmito I on porcine pancreatic phospholipase A2 activity was as potent as that of bovine lung annexin I. The presence of annexin I in mitochondria was confirmed by an electron-microscopic study. AXmito I was shown to be phosphorylated by intrinsic protein tyrosine kinases on its tyrosine residues. This annexin was also phosphorylated by protein kinase C. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 45 5 1991 Effects of beta-adrenergic blocking agents on specific binding of [3H]D-Ala2-Met5-enkephalinamide and [3H]naloxone. 295 299 EN Haruhiko Takayama Norio Ogawa Masato Asanuma Hiroshi Hirata Toshio Ogura Zensuke Ota Article 10.18926/AMO/32199 To gain further insight into the central nervous system (CNS)-action of beta-adrenergic blocking agents (beta-blockers), we examined the effects of various kinds of beta-blockers on opioid receptors (Op-Rs) using radiolabeled receptor assay (RRA). We demonstrated that beta-blockers are competitively bound to Op-Rs in the CNS. Sodium index of beta-blockers in [3H]naloxone binding study indicated that beta-blockers had the mixed agonist-antagonist activity of opiates. The relative potency of beta-blockers in opioid RRA was negatively correlated with their membrane stabilizing activity. Neither beta-blocking activity nor intrinsic sympathomimetic activity was correlated with IC50 values of beta-blockers in opioid RRA. While it is widely accepted that beta-blockers have a tranquilizing activity, a part of the tranquilizing action of beta-blockers may be mediated through Op-Rs in the CNS. Although beta-blockers may have effects on their own receptors (beta-receptors) in the CNS, the more precise mechanisms of central action of these drugs must be further investigated. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 58 1 2004 Effectiveness of a nonsteroidal anti-inflammatory drug for nocturia on patients with benign prostatic hyperplasia: a prospective non-randomized study of loxoprofen sodium 60 mg once daily before sleeping. 45 49 EN Tohru Araki Teruhiko Yokoyama Hiromi Kumon Article 10.18926/AMO/32115 We explored the effectiveness of loxoprofen sodium (loxoprofen), which is the most common non-steroidal anti-inflammatory drug (NSAID) in Japan, for patients with benign prostatic hyperplasia (BPH) complaining of nocturia. A total of 93 BPH patients aged 49-84 years were enrolled in the study. These patients had received standard drug therapy with alpha1-blocker for BPH, followed by anticholinergic drugs, hypnotics, tricyclic antidepressants, and/or antiduretic hormone, but they still complained about 2 or more episodes of nocturia. They each took a single 60-mg tablet of loxoprofen prior to sleeping at night for 14 days in addition to their BPH treatments. The effects were assessed by questionnaire before and after treatment as excellent (nocturia disappeared or decreased by 2 or more voids/night), improved (nocturia decreased by 1 void/night), unchanged, or worsened (nocturia increased). Nocturia improved or disappeared in 74.2% of patients: excellent, improved, unchanged, and worsened results were obtained in 37.6%, 36.6%, 21.5%, and 4.3% of patients, respectively. The effects were better in patients whose baseline nocturia was > 2 times than in those with a lesser frequency at enrollment (P = 0.04). Loxoprofen can be an effective and useful treatment option for patients with BPH complaining of refractory nocturia. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 33 3 1979 Properties of erythrocyte catalase from heterozygotes for Japanese type acatalasemia. 205 211 EN Masana Ogata Junko Mizugaki Article 10.18926/AMO/32073 The level of blood catalase activity in heterozygotes for Japanese type acatalasemia was demonstrated to be about half of normal levels by means of titration and spectrophotometric methods. A distribution plot of catalase activities in heterozygous blood was completely separate from that of normal blood. Comparative analysis of the partially purified erythrocyte catalase preparations obtained from normal and heterozygous individuals revealed no distinct differences between them regarding stability to heat, sodium dodecyl sulfate and some enzyme inhibitors or pH dependency. The erythrocyte catalase in heterozygotes for Japanese type acatalasemia contains about half the normal specific activity and as stable as that in normal individuals. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 55 2 2001 Severe fenitrothion poisoning complicated by rhabdomyolysis in psychiatric patient. 129 132 EN Koujiro Futagami Naofumi Hirano Emiko Iimori Kenichi Motomura Michiko Ide Yasuhumi Kataoka Hiroaki Araki Yutaka Gomita Ryozo Oishi Article 10.18926/AMO/32011 Non-traumatic rhabdomyolysis associated with organophosphate intoxication has not been generally reported. We report here in a severe case of fenitrothion poisoning complicated by rhabdomyolysis. A 43-year-old woman ingested approximately 100 ml of fenitrothion emulsion (50%) in an attempt to commit suicide. On day 3 after admission, her creatine phosphokinase (CPK) peaked at 47,762 IU/L. She received supportive treatment included sodium bicarbonate and fluid resuscitation. However, muscarinic symptoms including excessive miosis and salivation developed on day 5 when her CPK levels decreased. The delay in cholinergic symptoms might have been due to the trihexyphenidyl she took with the antipsychotic drugs. Fortunately, the present patient recovered from the acute cholinergic crisis, and acute renal failure was prevented by early diagnosis. This is a case of organophosphate poisoning complicated by rhabdomyolysis in a psychiatric patient. The masking of acute cholinergic symptoms should be taken into consideration in such patients. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 55 4 2001 Significance of adrenomedullin under cardiopulmonary bypass in children during surgery for congenital heart disease. 245 252 EN Mamoru Takeuchi Kiyoshi Morita Tatsuo Iwasaki Yuichiro Toda Katsunori Oe Naoyuki Taga Masahisa Hirakawa Article 10.18926/AMO/31995 To elucidate the effect of adrenomedullin (AM) on fluid homeostasis under cardiopulmonary bypass (CPB), we investigated the serial changes in plasma AM and other parameters related to fluid homeostasis in 13 children (average age, 28.2 months) with congenital heart disease during cardiac surgery under CPB. Arterial blood and urine samples were collected just after initiation of anesthesia, just before commencement of CPB, 10 min before the end of CPB, 60 min after CPB, and 24 h after operation. Plasma AM levels increased significantly 10 min before the end of CPB and decreased 24 h after operation. Urine volume increased transiently during CPB, which paralleled changes in AM. Simple regression analysis showed that plasma AM level correlated significantly with urinary vasopressin, urine volume, urinary sodium excretion, and plasma osmolarity. Stepwise regression analysis indicated that urine volume was the most significant determinant of plasma AM levels. Percent rise in AM during CPB relative to control period correlated with that of plasma brain natriuretic peptide (r = 0.57, P < 0.01). Our results suggest that AM plays an important role in fluid homeostasis under CPB in cooperation with other hormones involved in fluid homeostasis. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 56 4 2002 Improvements in the measurement of stool decay-accelerating factor in the detection of colorectal cancer. 171 176 EN Shogen Ohya Motowo Mizuno Mikihiro Kawada Junichirou Nasu Hiroyuki Okada Hiroyuki Shimomura Kazuhide Yamamoto Teizou Fujita Takao Tsuji Article 10.18926/AMO/31686 We have previously developed an enzyme-linked immunosorbent assay (ELISA) to measure stool decay-accelerating factor (DAF) and found that stool DAF concentrations were significantly elevated in patients with colorectal cancer, suggesting that the measurement of stool DAF may be valuable for the detection of colorectal cancer. In order to refine the assay for the measurement of stool DAF, we investigated 1) effects of centrifugation of stool samples, 2) effects of detergents, and 3) adequate combination of various anti-DAF monoclonal antibodies for the ELISA system using only monoclonal antibodies. We found that high-speed centrifugation could be omitted and that only the removal of large undigested food residues by centrifugation of short duration in a low-speed benchtop microcentrifuge sufficed to adequately prepare the stool samples. Addition of 2 detergents, octyl beta-glucoside and sodium deoxycholate, known to solubilize glycosyl-phosphatidylinositol-anchored proteins such as DAF, did not influence stool DAF values. By using 2 mouse anti-DAF monoclonal antibodies (clone 4F11 and 1C6), we were able to achieve a stable ELISA for the measurement of stool DAF using a uniform source of antibodies. The results should allow us to consistently apply the DAF assay for routine use in the detection of colorectal cancer. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 53 1 1999 Hydrocortisone Sodium Succinate Suppressed Production of Interleukin-10 by Human Peripheral Blood Mononuclear Cells: Clinical Significance 55 59 EN Hideo Kohka Hiromi Iwagaki Tadashi Yoshino Kenta Kobashi Shinnya Saito Hiroshi Isozaki Norihisa Takakura Noriaki Tanaka Article 10.18926/AMO/31646 Corticoids are well known for their immunosuppressive properties. Interleukin-10 (IL-10) is an intrinsic antiinflammatory peptide in immune diseases, originally identified as cytokine synthesis inhibitory factor. We examined the effect of hydrocortisone sodium succinate (HSS) on the production of IL-10 by human peripheral blood mononuclear cells (PBMCs). PBMCs from healthy volunteers and cancer-burden patients were preincubated separately with or without HSS for 1 h, then stimulated with 5 microg/ml lipopolysaccharide (LPS). Production of IL-10 by human PBMCs was detected with LPS stimulation and its production was higher in cancer-burden patients than in normal volunteers, although this was not statistically significant. HSS suppressed production of IL-10 by LPS-stimulated PBMCs in a dose-dependent manner both in normal volunteers and in cancer-burden patients. These results indicate that, in addition to their antiinflammatory properties, corticoids act to restore the immunosuppressive states even in cancer-burden states No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 53 5 1999 Evidence that the nitrergic neurotransmitter and endothelium-derived relaxing factor might be S-nitrosothiols in the mouse corpus cavernosum. 209 215 EN Kansu Buyukafsar Cemil Gocmen Ata Secilmis Yusuf Karatas Sinem Gokturk Nuri Ihsan Kalyoncu Article 10.18926/AMO/31632 The effects of thimerosal, a sulfhydryl oxidizing agent on nitrergic, endothelium-dependent and -independent relaxations were investigated to examine the possibility that the nitrergic neurotransmitter and endothelium-derived relaxing factor (EDRF) could be S-nitrosothiol or free nitric oxide (NO) in the isolated mouse corpus cavernosum. Thimerosal (5 x 10(-6)-2 x 10(-5) M) inhibited or almost abolished electrical field stimulation--(EFS, 30V, 0.5 ms, 15 sec, 1, 2, 4, 8, 16 Hz), acetylcholine--(ACh, 5 x 10(-8)-1.25 x 10(-6) M), glyceryl trinitrate--(GTN, 3 x 10(-7)-3 x 10(-6) M), and S-nitrosoglutathione--(GSNO, 5 x 10(-6)-1.25 x 10(-4) M) induced relaxations. Thiomerosal inhibition seems to be specific to L-arginine NO pathways since it had no effect on acidified sodium nitrite--(10(-4)-5 x 10(-4) M), photoactivated sodium nitrite--(2 x 10(-4) M), isoprenaline--(10(-6) M), or papaverine--(10(-4) M) elicited relaxations. Moreover, the inhibitory effect of thimerosal on the nitrergic, ACh- or GTN-induced relaxations were partly reversed by sulfhydryl-containing compounds, L-cysteine (10(-3) M), dithiothreitol (10(-3) M), or glutathione (10(-3) M). However L-methionine (10(-3) M), which contains a methyl group on the sulphur atom, failed to restore the thimerosal inhibition. Thimerosal did not change the contraction produced by 10(-4) M NG-nitro-L-arginine methyl ester. These findings indicate that the nitrergic neurotransmitter as well as EDRF may not be free NO but NO-transferring molecules, probably S-nitrosothiols, in the mouse corpus cavernosum. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 39 3 1985 Evaluation of Talbot's Safety Zone of Infusion Volume and Osmolality in Infusion Therapy for Decompensated Liver Cirrhosis 179 190 EN Shiro Yuasa Tatsuya Itoshima Ryosaku Ono Hideo Nagashima Article 10.18926/AMO/31531 Problems with infusion therapy for correcting fluid and sodium imbalance in decompensated liver cirrhosis (DLC) were investigated by establishing the safety zone of Talbot et al. for parenteral fluid therapy in 4 DLC patients infused with over 900 ml of fluid each day for at least 9 days. The safety zone was different in each case. The safe infusion volume decreased and the safe electrolyte concentration shifted to a lower osmolality when there was ascites with renal failure than ascites without renal failure. Infusion therapy was performed without deterioration of the water and sodium balance in those patients whose infusion volume and fluid osmolality were in the safety zone. In contrast, ascites retention increased and peripheral edema appeared in patients whose infusion volume and osmolality were out of the safety zone. Therefore, the safety zone should be determined repeatedly during infusion therapy. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 39 3 1985 Effects of pentazocine and concomitant clonidine on opioid receptors in the rat brain. 217 220 EN Norio Ogawa Yukiko Hirose Hiroo Kuroda Haruhiko Takayama Article 10.18926/AMO/31528 The changes in opioid receptors (Op-R) caused by repeated administration of pentazocine and the effect of concomitant clonidine were investigated. Binding of [3H] naloxone was markedly decreased in the absence of Na+, but was increased in the presence of Na+ in the diencephalon-mesencephalon of chronic pentazocine-treated rats. No significant changes were observed in the cerebral cortex of pentazocine-treated rats. The pentazocine-induced changes in Op-R were abolished by the concurrent use of clonidine, an alpha-adrenergic agonist, which has been shown to relieve the withdrawal symptoms of morphine. This result indicated that the behavioral action of clonidine can also be observed at the Op-R level. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 12 3 1958 An analytical study on the reduction of neotetrazolium chloride by the terminal electron transport system 193 204 EN Takuzo Oda Hiroaki Okazaki Article 10.18926/AMO/31363 In order to determine the steps with which the reaction of neotetrazolium chloride reduction conjugates in the terminal electron transport system, an analytical study on the neotetrazolium reduction by tissue homogenates was carried out using various substrates such as sodium succinate, p-phenylenediamine, sodium malate, sodium α-glutamate and DPN, and inhibitors such as sodium malonate, potassium cyanide and antimycin A, as the results the following conclusions were drawn. 1. The reaction of neotetrazolium reduction by tissue homogenate using sodium succinate as substrate is mainly the succinoxidase system reaction; and the reaction takes place conjugating about 50 per cent in the step of the succinic dehydrogenase system (succinic dehydrogease, cytochrome b and cytochrome C1), of these about 15 per cent conjugates in the step prior to the antimycin A sensitive step and 35 per cent in the step itself; and about 50 per cent in the step of cytochrome c oxidase. 2. In the case using p-phenylenediamine as substrate the reaction of neotetrazolium reduction is the reaction due to the activity of cytochrome c-cytochrome oxidase system; and when p-phenylenediamine is used with the sufficient amount of cytochrome c, the reaction appears to be dependent on cytochrome c oxidase activity. Neotetrazolium reduction in all these reactions takes place conjugating in the step of cytochrome c oxidase. 3. In the case where DPN and substrates taking DPN as a coenzyme are used, the reaction of neotetrazolium reduction is mainly the reaction conjugating at the step below antimycin A sensitive step in the DPNHcytochrome c reductase system (flavoprotein, cytochrome b and cytochrome c;), probably with the flavoprotein of DPNH-dehydrogenase. 4. Endogenous dehydrogenase reactions are the sum total reactions conjugating at the steps prior to the antimycin A sensitive step in the terminal electron transport system and with other various reduction systems which are not inhibited by antimycin A. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 12 2 1958 Effect of histamine releasers and of anti-inflammatory drugs on the egg-white edema of rat's hind paws in relation to skin histamine 93 111 EN Shozo Irino Article 10.18926/AMO/31359 1. A method was described for a fairly accurate judgement of the effect of drugs inhibiting the edema in hind paws of a rat caused by local injection of egg white. 2. The degree of inhibition of egg-white edema by single doses of sinomenine, compound 48/80, or dextran was in parallel with histamine reduction in skin and other tissues of the paws (and the skin of abdomen), although prevention of the edema by prolonged treatment with sinomenine was incomplete even when the releasable histamine of the skin was practically exhausted. 3. Sodium salicylate, aminopyrine, butazolidine sodium, cortisone, and guaiazulene were capable of inhibiting egg-white edema without modifying the content of skin histamine. These drugs and a small dose of phenergan potentiated the inhibition by dextran of egg-white edema and inhibited the release of histamine by dextran. These actions lasted for over 24 hours with the exception of guaiazulene. 4. Irgapyrin and a large dose of phenergan, which possess actions of histamine release and of histamine release inhibition and also antihistaminic action, caused a slight reduction of skin histamine and a comparatively marked inhibition of the edema. 5. In adrenalectomized or hypophysectomized rats, the edema-inhibiting effect of salicylate and aminopyrine decreased but that of cortisone increased. The effect of guaiazulene remained unchanged. 6. The observations that inhibition of egg-white edema is caused by (a) histamine releasers, (b) histamine-release inhibitor, and (c) drugs exerting both histamine release and inhibition of the release were discussed with the consideration to a relationship between egg-white edema and skin histamine. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 52 5 1998 Expression of amiloride-sensitive sodium channel in rat eye. 279 283 EN Toshihiko Matsuo Article 10.18926/AMO/31319 To study the expression of the amiloride-sensitive sodium channel, a putative mechano-receptor in the rat eye, reverse transcriptase-polymerase chain reaction and in situ hybridization were done. The gene for the alpha subunit of the amiloride-sensitive sodium channel was shown by polymerase chain reaction to be expressed in mRNA isolated from the whole eye tissue. In situ hybridization demonstrated that the gene was expressed in basal layers of the corneal and conjunctival epithelium, ciliary epithelial cells, lens epithelial cells at the equator, retinal and iris pigment epithelial cells, ganglion cells and cells in the inner and outer nuclear layers of the retina. The results suggest that the amiloride-sensitive sodium channel plays a role in maintaining sodium balance as well as in possible mechanosensation in these ocular tissues. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 52 6 1998 Can POSSUM, a Scoring System for Perioperative Surgical Risk, Predict Postoperative Clinical Course ? 325 329 EN Naoto Gotohda Hiromi Iwagaki Satoshi Itano Sadayuki Horiki Toshiya Fujiwara Shinya Saito Akio Hizuta Hiroshi Isozaki Norihisa Takakura Norihiko Terada Noriaki Tanaka Article 10.18926/AMO/31304 POSSUM, a Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity, is a scoring system which assesses perioperative surgical risks (Copeland GP et al.: Br J Surg, 1991, Vol 78, 356-360). The POSSUM scoring system consists of two categories of assessment to assess the risk of surgery. A 12-factor (age, cardiac status, pulse rate, systolic blood pressure, respiratory status, Glasgow Coma Score, serum concentration of urea, potassium and sodium, hemoglobin concentration, white cell count and findings on electrocardiography) and 4-grade physiological score (PS) were developed. This was combined with a 6-factor (type of surgical procedure, number of procedures, blood loss, peritoneal soiling, presence of malignancy and mode of surgery) and 4-grade operative severity score (OSS). The present paper attempts to validate it retrospectively. Postoperative hospitalization period and duration of antibiotics administration were both significantly correlated with OSS, but not with PS. These results suggest that the POSSUM scoring system is useful for predicting the postoperative clinical course. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 35 1 1981 A perifusion method for examining arginine vasopressin (AVP) release from hypothalamo-neurohypophyseal system. 27 35 EN Norihito Ohno Kozo Hashimoto Sho Yunoki Jiro Takahara Tadashi Ofuji Article 10.18926/AMO/31292 A perifusion method has been developed using rat hypothalamo-neurohypophyseal system (HNS) or neural lobe to investigate the control mechanism of arginine vasopressin (AVP) release. A specific radioimmunoassay (RIA) for AVP was developed to measure AVP in perifusion medium employing anti-AVP serum which was obtained by immunizing rabbits. At a final dilution of 1/12,000, the antiserum showed less than 0.66 and 0.01% cross reactivity with lysine-vasopressin and oxytocin, respectively. But it did not cross reacted with other peptide hormones. The lowest detectable level of vasopressin was 0.5 pg/tube. The intra-assay coefficient of variation averaged 10.4%. The dilution curve of perifused medium was well paralled to the standard curve of AVP assay. AVP release from HNS or neural lobe gradually declined to the stable level in 90-120 min after the initiation of perifusion. Good repeatability of the AVP release from neural lobe was recognized by repeated stimulation with 10 min perifusion of 60 mM KCl at every 60 min. HNS released AVP in dose related manner to the osmotic challenge of sodium or glucose, and AVP release was stimulated from HNS by prostaglandin E2, but not by dopamine. These results show that the perifusion methods using AVP-RIA is a useful method to examine the AVP release from HNS or neural lobe. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 35 3 1981 A scanning electron microscopic study of the two-step effect of cytochalasin B on Ehrlich ascites tumor cells. 197 204 EN Junzo Sasaki Masaaki Imanaka Sadahiro Watanabe Nagayasu Otsuka Shu Nakamoto Masaharu Mori Article 10.18926/AMO/31288 The effect of cytochalasin B (CB) on the surface structure of Ehrlich ascites tumor cells was investigated using the scanning electron microscope. The effect occurs in two steps: formation of zeiotic knobs on the cell surface and subsequent grouping of the knobs at one pole of the cell. The early step of zeiotic knob formation occurs at low concentrations of CB (0.5-1 microgram/ml) at 37 degrees C and at high concentrations of the drug (5-10 microgram/ml) at low temperature but within 1 min at 37 degrees C. This step is only partially inhibited by 5 x 10(-3) M sodium azide. The subsequent grouping of zeiotic knobs lasts for more than 2 min at 37 degrees C and occurs only in the case of high concentrations of CB. It is inhibited by sodium azide and is often associated with grouping of the microvilli, which are then lost from all of the cell surface except the area of knob-grouping. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 35 5 1981 Long-term preservation of transfecting activity of avian sarcoma proviral DNA. 377 379 EN Hajime Ogura Tazuko Fujiwara Brief Note 10.18926/AMO/31280 The integrated proviral DNA of avian sarcoma virus (ASV) in host cell chromosomes has been isolated and stored in saline sodium citrate (SSC) solution or in 70% ethanol at 4 degrees C in a refrigerator over 4 years. This DNA was assayed by transfection of chick embryo cells(CEC). The biological activity of cellular transformation by the stored DNA was compared with that of a fresh isolate of the proviral DNA. The efficiency of the transfection by each DNA was almost the same. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 42 6 1988 Effect of heart rate and myocardial contractile force on coronary resistance. 335 342 EN Daiji Saito Masayuki Ueeda Kazuyoshi Hina Hirofumi Watanabe Tsutomu Mima Masahiro Hasui Nobuyuki Yamada Shoichi Haraoka Takao Tsuji Article 10.18926/AMO/31008 The effect of the heart rate and myocardial contractile force on the extravascular resistance to blood flow of the left anterior descending coronary artery (LAD) was evaluated in 15 mongrel dogs anesthetized with sodium pentobarbital. The LAD was maximally dilated by intracoronary infusion of adenosine, which precluded the influence of vasomotor tone. Increases in the heart rate and myocardial contractile force decreased coronary blood flow in the absence of a change in coronary perfusion pressure. The changes in mean coronary resistance showed a significant linear relationship to changes in developed tension. The changes in coronary resistance caused by varying the heart rate and contractile force were so small that a normal coronary vascular tree could easily compensate for the increase in resistance. However, it is supposed that with critical stenosis of the vascular tree even a small increase in resistance might cause deleterious effects on coronary blood flow. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 62 6 2008 A Clinical Investigation of the Mechanism of Loxoprofen, a Non-steroidal Anti-inflammatory Drug, for Patients with Nocturia 373 378 EN Tohru Araki Teruhiko Yokoyama Motoo Araki Seiji Furuya Original Article 10.18926/AMO/30949 We previously reported the effectiveness of loxoprofen sodium (loxoprofen), a non-steroidal anti-inflammatory drug, for patients with lower urinary tract symptoms (LUTS) complaining of nocturia. In this study, we explored the mechanism of loxoprofen in the treatment of nocturia. Fifty-six patients complaining of nocturia were enrolled. They took a single 60-mg tablet of loxoprofen at bedtime for 14 days. The effects of this treatment were assessed by bladder diaries. Nocturia improved (nocturia decreased ≥1 void/night) in 40 patients (71.4%). Nocturnal urine volume was reduced in 31 of 40 (77.5%) without nocturnal single-void volume increase. Nocturnal single-void volume increased in 4 of 40 (10.0%) without nocturnal urine volume reduction. Two of 40 (5.0%) demonstrated both nocturnal urine volume reduction and nocturnal single-void volume increase. Three (7.5%) were exceptions to the above. In conclusion, the main mechanism of loxoprofen is the reduction of nocturnal urine volume for the treatment of nocturia and the second mechanism is the increased bladder capacity. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 29 1 1975 Microheterogeneity of rat alpha-fetoprotein. 29 32 EN Akiharu Watanabe Kazuhisa Taketa Kiyowo Kosaka Article 10.18926/AMO/30935 A purified and homogeneous preparation of rat alpha-fetoprotein (AFP) was separated into two components, AFPa and AFPb, by polyacrylamide gel electrophoresis. These two components had a definite difference in electrostatic net charge and gave only a single band on sodium dodecyl sulfate-electrophoresis. Neuraminidase-treated AFP gave clearly separable, slower moving four to six and finally two components depending on the time of incubation with neuraminidase. The time-dependent conversion of each AFPa and AFPb into slower migrating components upon neuraminidase treatment was confirmed by re-electrophoresis of separated and similarly treated AFPa and AFPb. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 29 6 1975 Effect of sodium dodecyl sulfate on immuno-electrosyneresis between normal human erythrocyte membrane and sera of systemic lupus erythematosus patients 421 429 EN Shigeru Arimori Shinya Shinozawa Kiyoshi Hiraki Article 10.18926/AMO/30912 An anti-membrane antibody was present in the sera of systemic lupus erythematosus patients in immunoelectrosyneresis with sodium dodecyl sulfate (SDS) solubilized erythrocyte membrane as antigen. The SDS bound to protein was detected by chromatography at 10(-3)M concentration under U.V. light, at 10(-5)M concentration by the distilled water spray method and at 10(-6)M concentration by using rosaniline hydrochloride colorimetry. SDS was removed from the membrane protein at a concentration of 10(-3)M by the first gel filtration of Sephadex G-25 column and at a concentration of 10(-6)M by rechromatography of the same column. More than 99% of SDS in the solubilized erythrocyte membrane was removed by gel filtration. The antigenicity was still positive in the refiltrated fractions of systemic lupus erythematosus patients. Therefore, all precipitates in the gels were antigen-antibody aggregates. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 43 2 1989 Rapid purification of squirrel monkey retrovirus-H major gag protein by high performance liquid chromatography. 127 129 EN Shogo Ikeda Ken Tsutsui Masao Hatsushika Sekiko Watanabe Takuzo Oda Article 10.18926/AMO/30869 The major gag protein (p34) of squirrel monkey retrovirus-H was purified in one chromatographic step by anion-exchange high performance liquid chromatography. The virus in a crude fraction was disrupted with Brij 35 in the presence of three kinds of protease inhibitors. The soluble virus lysate was injected into a Polyanion SI column, and p34 was eluted with a linear salt gradient. The recovery of the protein was about 60%. The purified p34 was nearly homogenous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 51 4 1997 cDNA cloning, sequence analysis and expression of a mouse 44-kDa nuclear protein copurified with DNA repair factors for acid-depurinated DNA 195 206 EN Yuko Nakagawa Sekiko Watanabe Kosuke Akiyama Altaf H Sarker Ken Tsutsui Hajime Inoue Shuji Seki Article 10.18926/AMO/30763 We purified a 44-kDa nuclear protein from salt-extract of permeable mouse ascites sarcoma cells in an effort to isolate factors involved in the repair of acid-depurinated DNA. It was copurified with a major AP endonuclease (APEX nuclease) by sequential column chromatography then further purified by sodium dodecyl sulphate-poly-acrylamide gel electrophoresis as a possible DNA repair support factor. Its partial amino acid sequences were determined, and a cDNA clone for the protein was isolated from a mouse T-cell cDNA library using long degenerate oligonucleotide probes deduced from the amino acid sequence. The complete nucleotide sequence of the cDNA (1.7 kilobases) was determined. Northern hybridization using this cDNA detected two transcripts: 1.8kb being the major one and 2.6 kb being the minor one. The complete amino acid sequence for the protein predicted from the nucleotide sequence of the cDNA indicates that the 44-kDa protein consists of 394 amino acids with a calculated molecular weight of 43,698. In tests performed thus far, the recombinant 44-kDa protein expressed in Escherichia coli has not expressed any repair-support activity. It remains to be analyzed whether the protein attains this activity after appropriate posttranslational modifications. Most parts of the 44-kDa protein cDNA and the deduced amino acid sequence were found to be identical to those of the protein p38 -2G4, recently reported as a cell cycle-specifically modulated nuclear protein of 38kDa. The p38-2G4 may be a truncated form of the present 44-kDa protein. No potential conflict of interest relevant to this article was reported.