Expression dynamics of HAND1/2 in in vitro human cardiomyocyte differentiation

転写因子HAND1とHAND2の発現パターンと役割の解明 --ヒトiPS細胞から増殖能の高い心筋細胞を回収する--. 京都大学プレスリリース. 2021-07-27.Three genes determine heart cell growth. 京都大学プレスリリース. 2021-07-27.Hand1 and Hand2 are transcriptional factors, and knockout mice of these genes show left and right ventricular hypoplasia, respectively. However, their function and expression in human cardiogenesis are not well studied. To delineate their expressions and assess their functions in human cardiomyocytes (CMs) in vitro, we established two triple-reporter human induced pluripotent stem cell lines that express HAND1[mCherry], HAND2[EGFP] and either MYH6-driven iRFP670 or tagBFP constitutively and investigated their expression dynamics during cardiac differentiation. On day 5 of the differentiation, HAND1 expression marked cardiac progenitor cells. We profiled the CM subpopulations on day 20 with RNA sequencing and found that mCherry+ CMs showed higher proliferative ability than mCherry− CMs and identified a gene network of LEF1, HAND1, and HAND2 to regulate proliferation in CMs. Finally, we identified CD105 as a surface marker of highly proliferative CMs

Am80, a retinoic acid receptor agonist, activates the cardiomyocyte cell cycle and enhances engraftment in the heart

ヒトiPS細胞由来心筋細胞の生着能改善に向けた新しい方法. 京都大学プレスリリース. 2023-07-14.Am80, a retinoic acid receptor agonist, activates cell cycle in induced cardiomyocytes and enhances heart tissue engraftment. 京都大学プレスリリース. 2023-07-14.Human induced pluripotent stem cell-derived (hiPSC) cardiomyocytes are a promising source for regenerative therapy. To realize this therapy, however, their engraftment potential after their injection into the host heart should be improved. Here, we established an efficient method to analyze the cell cycle activity of hiPSC cardiomyocytes using a fluorescence ubiquitination-based cell cycle indicator (FUCCI) system. In vitro high-throughput screening using FUCCI identified a retinoic acid receptor (RAR) agonist, Am80, as an effective cell cycle activator in hiPSC cardiomyocytes. The transplantation of hiPSC cardiomyocytes treated with Am80 before the injection significantly enhanced the engraftment in damaged mouse heart for 6 months. Finally, we revealed that the activation of endogenous Wnt pathways through both RARA and RARB underlies the Am80-mediated cell cycle activation. Collectively, this study highlights an efficient method to activate cell cycle in hiPSC cardiomyocytes by Am80 as a means to increase the graft size after cell transplantation into a damaged heart

Rainstorm and Flood Disasters: Case study of Shelter Management by Manila Municipal DRR (Disaster Risk Reduction) Teams during Tropical Storm Mario (Fung Wong) in 2014

Floods are the leading cause of natural disaster-related deaths worldwide. This study aimed at assessing disaster preparedness and response by local Barangay (municipal) DRR (disaster risk reduction) teams during the flooding caused by tropical storm Mario in Manila in 2014. A cross-sectional and descriptive study was conducted, consisting of interviews in which five shelter managers participated. In total, 325 evacuees were received in 4 evacuation centers, whereas the remaining shelter received 30 families housed in tents. Only 3 shelters (60%) had some food and non-food items available prior to the arrival of evacuees. WASH (Water, sanitation and hygiene) services were insufficient; latrines were available in 4 (80%) shelters, but no latrine was available for displaced people housed in tents. Only 3 (60%) shelters had toilets cleaned regularly. Detergents, toothbrushes and toothpastes were provided in 3 (60 %) shelters, whereas only 2 (40%) had diapers for babies and none (0%) had hygienic period items for ladies. Food items were daily distributed in 3 (60%) shelters. Health services were not satisfactory, as medical consultations were organized but irregularly in 3 (60%) of the five shelters. Disaster preparedness and response in Barangay shelters were not satisfactory, suggesting the necessity for the central government to support local DRR volunteer teams. It is recommended to provide the volunteers with an inexpensive ICT (information and communications technology) tool to collect disaster preparedness data so that relief efforts will be more than sufficient at the time of disaster

Comparison of Self-Evaluations of Midwifery Practical Skills by Students in Graduate and Undergraduate Midwifery Programs

本研究の目的は、助産師養成課程で学んだ大学院生と大学生の妊婦健康診査実習前後の助産実践能力の到達レベルの違いを自己評価とその理由から明らかにすることである。　研究方法は、助産師養成課程の大学院生 10 名と大学 4 年生 9 名を対象に、妊婦健康診査での助産実践能力に対する自己評価と自己評価の理由について、実習前後に調査した。分析は、時期（実習前と実習後）と教育課程（大学院と学部）を二要因とし、自己評価項目を従属変数として二元配置分散分析と行った。また、自己評価の理由は最小単位に細分化し共通内容をまとめた。　結果として、妊婦健康診査実習前後の実践能力に対する自己評価得点で、教育課程で有意差が見られたのは「妊娠の経過予測」と「親準備」、実習前と実習後で有意差が見られたのは「胎位」「胎児の健康上状態」「胎児の成長」「妊娠経過の診断」「妊婦の訴えに対する助言」と健康教育ケアプランの「分娩準備」の 6 項目で実習後に自己評価得点は高くなっていた。交互作用で有意差が見られた「胎向」は、実習前に比べ実習後の自己評価得点が高いことに教育課程が関連していた。自己評価の理由から大学院生と学部生に共通のカテゴリー＜診察技術能力評価の根拠＞、＜妊娠経過の診断能力評価の根拠＞、＜ケア計画・実践評価の根拠＞、＜妊婦とのコミュニケーション能力評価の根拠＞、＜学びを深めた背景＞の 5 つが抽出されたが、コードは 1 つを除き異なっていた。　学部生の自己評価得点とその理由からは、助産師教育卒業時の「少しの助言で自立してできる」の到達レベルまで至っていなかったが、大学院生からは、助産診察技術の基本となる「胎向」は実習後に有意に高くなっており、自己評価の理由からも「少しの助言で自立してできる」の到達レベルにあったと示唆された。しかし、アセスメント能力、コミュニケーション能力と助産師としての専門性を得るためには、さらなる実習が必要であるという示唆が得られた。In this study, we surveyed graduate and undergraduate students in midwifery programs before and after undergoing antenatal examination clinical practice training. Students were asked for their own evaluations of how much the clinic had benefited their practical competence, and their reasons for their self-evaluations.Questionnaires were given to ten graduate students and nine undergraduates in 4-year university programs before and after their clinical practice. The two parameters independent variables were the timing of the survey （before and after clinical practice） and curriculum （graduate school and undergraduate）. The dependent variables were the 52 self-assessment items in the questionnaire. The variables were examined along the two dimensions by analysis of variance. The reasons the respondents gave for their ratings were arranged by minimum units and categorized. With regard to self-evaluation of practical skills before and after antenatal examination clinical practice, when only the main effect of group was examined, “prediction of course of pregnancy” and “preparation to be a parent” were significant. When only the main effect of time was examined, the following six items were significant: “fetal presentation,” “fetal health status,” “fetal growth,” “diagnosis of pregnancy course,” “advice for pregnant womenʼs appeal,” and “health plan to prepare for delivery.” When only interaction was examined, only “fetal position” was significant. The responses regarding the subjectsʼ reasons for their ratings were arranged into the following five categories: （1） basis for evaluation of examination skills; （2） basis for evaluation of ability to diagnose pregnancy progress; （3） basis for evaluation of ability to make and implement care plans; （4） basis for evaluation of ability to communicate with clients; and （5） factors contributing to deepening of the learning experience. Among these five categories, only one involved a criterion that was cited by both classes of students. In all other cases, the respective classes of students cited different criteria as reasons for their evaluations. The self-evaluations by undergraduate students suggested that upon completion of their midwifery course they felt that they were not quite at the level where they could handle the work on their own, but still required further advice from seniors. Graduate students tended to show more confidence in their own competencies in examination, gained from the practical experience of ongoing antenatal examinations of a number of women. However, this study suggested that further practical experience is necessary to obtain assessment and connection with their clients as midwives

Intracellular calcium level is an important factor influencing ion channel modulations by PLC-coupled metabotropic receptors in hippocampal neurons

Signaling pathways involving phospholipase C (PLC) are involved in various neural functions. Understanding how these pathways are regulated will lead to a better understanding of their roles in neural functions. Previous studies demonstrated that receptor-driven PLCβ activation depends on intracellular Ca2+ concentration ([Ca2+]i), suggesting the possibility that PLCβ-dependent cellular responses are basically Ca 2+ dependent. To test this possibility, we examined whether modulations of ion channels driven by PLC-coupled metabotropic receptors are sensitive to [Ca2+]i using cultured hippocampal neurons. Muscarinic activation triggered an inward current at -100 mV (the equilibrium potential for K+) in a subpopulation of neurons. This current response was suppressed by pirenzepine (an M1-preferring antagonist), PLC inhibitor, non-selective cation channel blocker, and lowering [Ca 2+]i. Using the neurons showing no response at -100 mV, effects of muscarinic activation on K+ channels were examined at -40 mV. Muscarinic activation induced a transient decrease of the holding outward current. This current response was mimicked and occluded by XE991, an M-current K+ channel blocker, suppressed by pirenzepine, PLC inhibitor and lowering [Ca2+]i, and enhanced by elevating [Ca 2+]i. Similar results were obtained when group I metabotropic glutamate receptors were activated instead of muscarinic receptors. These results clearly show that ion channel modulations driven by PLC-coupled metabotropic receptors are dependent on [Ca2+]i, supporting the hypothesis that cellular responses induced by receptor-driven PLCβ activation are basically Ca2+ dependent. © 2013 Elsevier B.V

HER2-Positive Metaplastic Breast Cancer with Resistance to Neoadjuvant Chemotherapy: Case Report

Introduction: Metaplastic breast carcinoma (MBC) is a rare histologic subtype of breast carcinoma, which is usually negative for estrogen receptor, progesterone receptor, and HER2. HER2-positive MBC is therefore extremely rare. Most MBCs have poor response to chemotherapy. HER2-targeted neoadjuvant chemotherapy (NAC) is widely performed and has high efficacy in treating HER2-positive breast cancer. We report an atypical case of HER2-positive breast cancer that had poor response to NAC and was diagnosed with MBC after the surgery. Case Presentation: A 73-year-old woman noticed a mass in her right breast and visited our hospital. The mass was diagnosed as hormone receptor-negative, HER2-positive invasive ductal carcinoma, T2N0M0 stage IIA. She received HER2-targeted NAC comprising trastuzumab + pertuzumab + docetaxel. Despite three courses, we observed disease progression. The next NAC regimen was composed of two courses of epirubicin + cyclophosphamide, but the cancer continued to grow. She stopped receiving NAC and underwent a unilateral mastectomy and sentinel lymph node biopsy. Although the preoperative pathological result of core needle biopsy specimen showed invasive ductal carcinoma, the postoperative pathological result of the surgical specimen was MBC. Conclusion: In this case, when the patient had undergone three courses of trastuzumab + pertuzumab + docetaxel, it would have been appropriate to review the result of the core needle biopsy with pathologists or to perform vacuum-assisted breast biopsy. This case suggests the importance of considering the possibility of special histologic subtypes such as MBC when a tumor with the diagnosis of invasive ductal carcinoma is resistant to NAC

Three dimensional motion analyses for rehabilitation version of Awa Odori exercise and the expectancy of physical effects

‘Awa Odori Exercise -Rehabilitation version- was developed in 2006 for the new trial of physical exercise for the aging and the impaired person with lower balance performance in Tokushima prefecture, Japan. Public relations of this exercise had been spreading over Tokushima since then. The characteristics of the exercise were highly familiar with most of people in Tokushima because of popularity in original ‘Awa Odori’. This study proposed the efficacies of Awa Odori Exercise as a rehabilitation exercise. This exercise expected the flexible balance reinforcements and the substitution for walking training with prevention of fall, bedridden and participating restriction for the old people, also promoting the health in Tokushima

A randomized phase III study of short-course radiotherapy combined with Temozolomide in elderly patients with newly diagnosed glioblastoma; Japan clinical oncology group study JCOG1910 (AgedGlio-PIII)

BACKGROUND: The current standard treatment for elderly patients with newly diagnosed glioblastoma is surgery followed by short-course radiotherapy with temozolomide. In recent studies, 40 Gy in 15 fractions vs. 60 Gy in 30 fractions, 34 Gy in 10 fractions vs. 60 Gy in 30 fractions, and 40 Gy in 15 fractions vs. 25 Gy in 5 fractions have been reported as non-inferior. The addition of temozolomide increased the survival benefit of radiotherapy with 40 Gy in 15 fractions. However, the optimal regimen for radiotherapy plus concomitant temozolomide remains unresolved. METHODS: This multi-institutional randomized phase III trial was commenced to confirm the non-inferiority of radiotherapy comprising 25 Gy in 5 fractions with concomitant (150 mg/m2/day, 5 days) and adjuvant temozolomide over 40 Gy in 15 fractions with concomitant (75 mg/m2/day, every day from first to last day of radiation) and adjuvant temozolomide in terms of overall survival (OS) in elderly patients with newly diagnosed glioblastoma. A total of 270 patients will be accrued from 51 Japanese institutions in 4 years and follow-up will last 2 years. Patients 71 years of age or older, or 71-75 years old with resection of less than 90% of the contrast-enhanced region, will be registered and randomly assigned to each group with 1:1 allocation. The primary endpoint is OS, and the secondary endpoints are progression-free survival, frequency of adverse events, proportion of Karnofsky performance status preservation, and proportion of health-related quality of life preservation. The Japan Clinical Oncology Group Protocol Review Committee approved this study protocol in April 2020. Ethics approval was granted by the National Cancer Center Hospital Certified Review Board. Patient enrollment began in August 2020. DISCUSSION: If the primary endpoint is met, short-course radiotherapy comprising 25 Gy in 5 fractions with concomitant and adjuvant temozolomide will be a standard of care for elderly patients with newly diagnosed glioblastoma. TRIAL REGISTRATION: Registry number: jRCTs031200099 . Date of Registration: 27/Aug/2020. Date of First Participant Enrollment: 4/Sep/2020

Relationship between Barthel Index scores during the acute phase of rehabilitation and subsequent ADL in stroke patients

The Barthel Index (BI) cannot be used to measure initial stroke severity or by extension, to stratify patients by severity in acute stroke trials because most patients are bedbound in the first few hours after stroke, either by their deficit or by medical directive. Our objectives were to clarify the threshold of acute BI for use in the prediction of subsequent independence in activities of daily living (ADL) and to assist in the definition of acute stroke rehabilitation goals. Subjects comprised 78 patients out of 191 inpatients admitted with acute stroke at our hospital during 2006-2007. The BI ADL score was divided into 2 ranges (BI≧60 and≦40), in a process similar to previous studies. During the acute period (from onset to approximately 3 weeks), all patients with a BI≧40 could improve their ADL in 6 months. Patients with a BI≦40 exhibited two ADL recovery outcomes (improved and no change) at 6 months. We also found that the skill level of basic activities related to standing was significant indicator of BI improvement (P<0.001). BI scores determined at approximately 3 weeks were reliable predictors of ADL disabilities at 6 months