Biochemical analysis of packing and assembling heptad repeat motifs in the coronavirus spike protein trimer.

During a coronavirus infection, the spike protein undergoes sequential structural transitions triggered by its receptor and the host protease at the interface between the virus and cell membranes, thereby mediating membrane fusion. After receptor binding, the heptad repeat motif (HR1/HR2) within the viral spike protein bridges the viral and cellular membranes; however, the intermediate conformation adopted by the spike protein when drawing the viral and cellular membranes into close proximity remains unclear due to its transient and unstable nature. Here, we experimentally induced conformational changes in the spike protein of a murine coronavirus by incubating the virus with its receptor, followed by exposure to trypsin. We then treated the virus/receptor complex with proteinase K to probe the tightly packed core structure of the spike protein. The conformations of the spike protein were predicted from the sizes of the protease digestion products detected by western blot analysis. Upon receptor binding, two bands (each showing different reactivity with a fusion-inhibiting HR2-peptide) were detected; we propose that these bands correspond to the packed and unpacked HR1/HR2 motifs. After trypsin-mediated triggering, measurement of temperature and time dependency revealed that packing of the remaining unpacked HR1/HR2 motifs and assembly of three HR1 motifs in a trimer occur almost simultaneously. Thus, the trimeric spike protein adopts an asymmetric-unassembled conformation after receptor binding, followed by direct assembly into the post-fusion form triggered by the host protease. This biochemical study provides mechanistic insight into the previously unknown intermediate structure of the viral fusion protein.IMPORTANCEDuring infection by an enveloped virus, receptor binding triggers fusion between the cellular membrane and the virus envelope, enabling delivery of the viral genome to the cytoplasm. The viral spike protein mediates membrane fusion; however the molecular mechanism underlying this process is unclear. This is because using structural biology methods to track the transient conformational changes induced in the unstable spike trimer is challenging. Here, we harnessed the ability of protease enzymes to recognize subtle differences on protein surfaces, allowing us to detect structural differences in the spike protein before and after conformational changes. Differences in the size of the degradation products were analyzed by western blot analysis. The proposed model explaining the conformational changes presented herein is a plausible candidate that provides valuable insight into unanswered questions in the field of virology.

Effect of an Enteral Formula Enriched with ω-3 Fatty Acids, Carnitine, and Vitamin D on Body Weight, Heart Weight, and Blood Biochemical Parameters in a Dahl Rat Heart Failure Model.

Malnutrition is known to worsen the prognosis of chronic heart failure. To gain information that may be helpful in establishing appropriate nutritional interventions for chronic heart failure, the present study was performed to investigate the efficacy of nutritional management with two enteral formulas, EH, with a standard nutritional composition, and ER, fortified with omega-3 fatty acids, vitamin D, and carnitine. Experiments were performed in a Dahl rat heart failure model. After being fed a standard rodent feed (MF) containing 8% NaCl (high salt-MF [HS-MF]) from 6 to 11 weeks of age, rats were assigned to freeze-dried EH or ER diets with an NaCl concentration of 8% (HS-ER or HS-EH) until 18 weeks of age. Serum albumin was significantly higher at 14 and 17 weeks of age in rats fed the HS-ER and HS-EH diets compared with those remaining on the HS-MF diet. Body weight was also significantly higher at 14 and 17 weeks of age in animals fed the HS-ER diet, showing that nutritional deterioration was prevented. Additionally, heart weight was significantly lower at 18 weeks of age in the HS-ER group than that in the HS-MF group, suggesting that cardiac hypertrophy was prevented. This study demonstrated improved nutritional status in a heart failure model in Dahl rats presumably owing to differences in nutritional composition in the diets. Future studies are needed to explore optimal nutritional management with enteral formulas in patients with chronic heart failure.

Health and environmental risk assessment of mercury in outdoor and indoor dust in artisanal and small-scale gold mining area in Amansie-west district in the Ashanti Region of Ghana.

Mercury (Hg) pollution around artisanal and small-scale gold mining (ASGM) areas has been of much concern. Many studies have reported elevated Hg concentrations in environmental media, but studies on dust relating to inhalation exposure of Hg around ASGM area are limited. In this study, we investigated Hg in indoor and outdoor dust to reveal environmental and human health risk around ASGM in Amansie West district, Ghana. Indoor and outdoor dust samples were collected from Manso Abore and Manso Nkwanta in Ashanti Region. Concentration of Hg in the samples were analyzed using a direct Hg analyzer. The mean and median value of Hg concentration in the indoor dust (n = 31) were 2.2 ± 3.6 mg/kg and 0.72 mg/kg respectively while that of the outdoor dust (n = 60) were 0.19 ± 0.48 mg/kg and 0.042 mg/kg, respectively. The mean and median Hg concentration in indoor dust were about 11 and 17 times higher respectively than that in the outdoor dust. The Hg concentration in the indoor dust was statistically significantly higher than that of the non-miner in Manso Abore (p < 0.05) but was not significant in Manso Nkwanta, probably due to higher mining activity. The geo-accumulation index of the outdoor dust ranged from unpolluted to extremely polluted while that of the indoor dust ranged from moderately polluted to extremely polluted. Health risk assessments suggested that there was no potential non-carcinogenic health effect for Hg exposure relating to the dust to residents living in rooms of miners and non-miners.

An international interdisciplinary approach to developing video educational materials promoting mental health literacy among adolescents in the Philippines.

AIM: Adolescent mental ill-health is a common international challenge affecting both high- and lower-middle-income countries. The Republic of the Philippines enacted its first mental health law in 2019, underlining the importance of the promotion of adolescent mental health education in schools. In Japan, course instructions about mental ill-health were formulated in a Course of Study that reflects governmental curriculum guidelines. Embedded since 2022, the Course of Study aimed to promote an understanding of current issues of adolescent health. The National Center for Global Health and Medicine in Japan has been sharing experience of promoting public health practices and advancing medical technology in low- and middle-income countries, especially in the Western Pacific region. This paper describes the development process and content of these resources by an interdisciplinary team from Japan and the Philippines. METHODS: The interdisciplinary team created an embedded mental health education programme using animated videos for the Philippine school curriculum to improve mental health literacy in adolescents. RESULTS: Two six-minute animated videos of age-relevant stories were created. The animation scenarios illustrate: 1) mental health problems and recovery; and 2) major symptoms of mental illnesses, including depression, anxiety, and schizophrenia. Each animated video presents several items for discussion among students and/or teachers, both in class and online for students unable to attend school. CONCLUSION: Our efforts may provide good opportunities for enhancing adolescents' mental health activities in the Philippines. In the following steps, we should investigate the effectiveness of school-based MHL using these animated videos with rigorous evaluation methods.

A Male Japanese Patient with Temple Syndrome Complicated by Type 2 Diabetes Mellitus.

We herein present the case of a 21-year-old male Japanese diabetic patient with Temple syndrome, caused by maternal uniparental disomy of chromosome 14. The patient was overweight and had type 2 diabetes, dyslipidemia, metabolic dysfunction-associated steatotic liver disease, and microalbuminuria. He had an increased fat mass in the truncal region and a decreased lean mass throughout the body. This may lead to insulin resistance due to the absence of delta-like homolog 1 (DLK1) and retrotransposon gag-like 1 (RTL1). The patient had experienced social withdrawal at home (hikikomori in Japanese), had poorly controlled type 2 diabetes, and was overweight despite receiving diet therapy and oral hypoglycemic agents.

Evaluating the cross-cultural competence instrument for healthcare professionals (CCCHP) among nurses in Okinawa, Japan.

BACKGROUND: To provide better quality healthcare services to patients with different linguistic and cultural backgrounds, the cross-cultural competence of medical professionals is important. However, assessing and improving the cross-cultural competence of healthcare professionals is difficult in Japan, as there is no standardized scale to measure the competence. This study's purpose was to translate the Cross-Cultural Competence instrument for Healthcare Professionals (CCCHP), which was developed and used in Europe, and to examine its reliability and validity among Japanese nurses. METHODS: During June and July 2021, nursing staff were invited to take web- and paper-based surveys in Okinawa Japan. The CCCHP (five-factor model with 27 items across motivation, attitude, skills, emotion, and knowledge) was translated using a combination translation method, and a five-point Likert scale was used for responses. Exploratory and confirmatory factor analyses and known-group method were used to examine structural validity, while Cronbach's alpha coefficient was used to test reliability. RESULTS: A total of 294 responses were analyzed; 77.2% had more than five years of experience. Since the fit index indicated that the five-factor model was not a good fit, it was modified to a four-factor model (J-CCCHP24) by moving three variables, removing the knowledge factor, and using the error covariance of the variables. The fit index after the modification was improved to comparative fit index (CFI) = 0.92, Tucker-Lewis index (TLI) = 0.91, root mean square error of approximation (RMSEA) = 0.05, and standardized root mean square residual (SRMR) = 0.06, and Cronbach's alpha was 0.85. The mean scores of J-CCCHP24 were significantly higher in the group with a history of overseas travel, higher foreign language skill, training in intercultural care, experience of foreign patient care, and intercultural interactions outside the workplace than in the group without these characteristics. CONCLUSION: This study confirmed the validity and reliability of the modified Japanese version of the CCCHP (four-factor model with 24 items). The results suggest that the exposure to different cultures on a personal level may help improve nurses' cross-cultural competence. Further refinement of this scale for practical use would encourage the implementation of necessary countermeasures to improve the cross-cultural competence of Japanese healthcare professionals.

Knowledge, attitudes, and practices related to dengue among public school teachers in a Central Luzon Province in the Philippines: an analytic cross-sectional study.

BACKGROUND: Dengue has become a major health issue in tropical regions as the numbers of reported cases and estimated infections continuously increase. In the Philippines, many challenges remain in preventing and controlling the disease amidst all the mitigation efforts of the government. This study sought to measure the health literacy of Filipino teachers and determine the associations among teachers' knowledge, attitudes, and selected practices (KAP) against dengue. METHODS: Elementary and secondary school teachers from the consistently declared dengue hotspots in the City of San Fernando, Pampanga, Philippines, from the years 2017 to 2019 were selected as target participants in this cross-sectional study. A self-administered online survey tool was used in this study for both screening of participants and the KAP survey proper. STATA, descriptive statistics, and multiple logistic regression were used for the data analysis. Odds Ratios (ORs) and 95% confidence intervals (CIs) were reported. RESULTS: The study comprised 604 participants whose mean age was 38.4 years. Television was determined as the top media source of information, and various health staff were the most trusted and common source of information. Good knowledge on dengue treatment (OR = 1.81; 95% CI 1.18-2.78) and dengue prevention (OR = 1.85; 95% CI 1.26-2.71) were positively associated with having good practices on protection against mosquito bites. Good knowledge on dengue signs and symptoms (OR = 1.56; 95% CI 1.02-2.37) and dengue prevention (OR = 2.38; 95% CI 1.59-3.58) were positively associated with having good practices on preventing breeding sites. Those with positive perceived susceptibility to dengue had lower odds of having good practices on protection against mosquito bites (OR = 0.64; 95% CI 0.41-0.99) and of having good practices on preventing breeding sites (OR = 0.46; 95% CI 0.26-0.81). CONCLUSION: Even with the existing dengue policies, programs, and strategies, and the high disease literacy rate of Filipinos, dengue remains a struggle with an increasing case rate. Therefore, specific concepts should be emphasized, and interventions should be fine-tuned to better reach and influence the target population to attain a dengue-free Philippines.

Multi-country case study on school health policy and its implementation in relation to COVID-19 control in Micronesia Small Islands Developing States.

INTRODUCTION: The COVID-19 pandemic caused school closures and rises in mental illness and non-communicable disease among school children worldwide. The Pacific Small Islands Developing States (SIDS) were also affected, but school health activities, which can effectively reduce negative effects of COVID-19, were not widely implemented compared to other Asia-Pacific countries. This study examined current school health implementation and related policies at national, local, and school levels in the Micronesia SIDS according to phases of COVID-19 control. METHODS: Multi-country case study targeted the Federated States of Micronesia (FSM), Republic of the Marshall Islands (RMI), and Republic of Palau (ROP). These studies focused on school health implementation periods according to the PPR (Prevention, Preparedness, and Response) concept: Phase #1: prevention/preparedness, #2: early phase response, and #3: chronic phase response/recovery phase. Data were collected through policy document reviews that identified school health policies related to COVID-19 controls in the three phases and key informant interviews (KIIs) with 44 key informants (FSM, n = 14; RMI, n = 18; ROP, n = 12) whose work related to school health. The collected data were analyzed using content analysis methods according to the conceptual framework in this study. RESULTS: This study identified three factors of school health implementation related to COVID-19 controls: promotion of decentralized education (FSM), implementation of COVID-19 controls in the school community (RMI), and disaster management for the protection of students including response to infectious disease (ROP). In Phase #1, no country had established a school health policy. In Phase #2, three enablers were identified in FSM and ROP, as reflected in COVID-19 controls by the education and health sectors. In Phase #3, RMI implemented COVID-19 controls in the school community. Documents on youth policy and disaster management in ROP were updated to reflect the chronic phase response and response to future public health crises. CONCLUSIONS: A decentralized education was instrumental in immediately implementing COVID-19 control measures in schools at national and local levels for coordination between education and health sectors. Despite each county's multi-sectoral approach to engage COVID-19 controls in schools, local government organization requires strengthening and implementation of the formulated school health policy. In preparation for the next public health crisis, school health should be promoted that is integrated into both infection control and disaster management.

Leadership of school principals for school health implementation among primary schools in Mataram, Indonesia: a qualitative study.

BACKGROUND: Health-promoting schools (HPS) are acknowledged as a comprehensive approach to improving children's health and educational outcomes through learning and school life. Principals are key players in HPS implementation. However, concrete leadership practices in HPS in low- to middle-income countries have not been clarified. Therefore, this study aimed to explore and generate themes surrounding the leadership practices of principals in implementation of school health in Indonesia, a predominantly Muslim country consisting of diverse religions that have expanded HPS at the national level. METHODS: In-depth interviews and focus group discussions (FGDs) were conducted with the principals, teachers, parent representatives, and school board committee members in 10 target schools. FGDs were conducted with school health supervisory board members in Mataram City. All interviews were recorded and transcribed. Thematic analysis was undertaken to generate themes. RESULTS: The principals demonstrated leadership practices based on their religious beliefs, values, and morals. This may suggest that beliefs and morals support an understanding of their responsibility to ensure the well-being of all school community members, regardless of religion, in a diverse environment that is predominantly Muslim but also multicultural and multi-religious. Further, these beliefs and morals might reinforce implementation of school health. Importantly, the principals' coordination skills in cooperating with multiple sectors might contribute to successful implementation of school health. Also, principals emphasized they were tasked to develop capacity for implementation of school health. Thus, principals should understand leadership in the implementation of school health as their duty from the training stage to encourage health at the school level. CONCLUSION: In this study, "have professional educators' beliefs and religious beliefs and Indonesia's morals" was generated as a new theme, whereas several common themes were found as in previous studies. The results of this study suggested the importance of leadership by principals in the implementation of school health. Strengthening the capacity of school principals by integrating the contents of school health leadership practice into pre- and in-service training through the development of a policy on principals' duties in school health might contribute to the successful implementation of school health.

Terminus-Selective Covalent Immobilization of Heparin on a Thermoresponsive Surface Using Click Chemistry for Efficient Binding of Basic Fibroblast Growth Factor.

Cell therapy using endothelial cells (ECs) has great potential for the treatment of congenital disorders, such as hemophilia A. Cell sheet technology utilizing a thermoresponsive culture dish is a promising approach to efficiently transplant donor cells. In this study, a new method to prepare terminus-selective heparin-immobilized thermoresponsive culture surfaces is developed to facilitate the preparation of EC sheets. Alkynes are introduced to the reducing terminus of heparin via reductive amination. Cu-catalyzed azide-alkyne cycloaddition (CuAAC) facilitates efficient immobilization of the terminus of heparin on a thermoresponsive surface, resulting in a higher amount of immobilized heparin while preserving its function. Heparin-immobilized thermoresponsive surfaces prepared using CuAAC exhibit good adhesion to human endothelial colony-forming cells (ECFCs). In addition, upon further binding to basic fibroblast growth factor (bFGF) on heparin-immobilized surfaces, increased proliferation of ECFCs on the surface is observed. The confluent ECFC monolayer cultured on bFGF-bound heparin-immobilized thermoresponsive surfaces exhibits relatively high fibronectin accumulation and cell number and detaches at 22 °C while maintaining the sheet-like structure. Because heparin has an affinity for several types of bioactive molecules, the proposed method can be applied to facilitate efficient cultures and sheet formations of various cell types.

Bacteriological and molecular characterization of temperature- and CO2-dependent Streptococcus pneumoniae serotype 24F ST162 isolated from Japanese children.

IMPORTANCE: We characterized Streptococcus pneumoniae serotype 24F sequence type (ST) 162 isolated from Japanese children with invasive pneumococcal disease (IPD). Owing to its highly invasive nature, serotype 24F is expected to be isolated from clinically significant cases. Serotype 24F ST162 isolates tested in the present study did not grow at 35°C in ambient air. Therefore, antimicrobial susceptibility testing using the broth microdilution method, which is usually conducted in ambient air, cannot be performed, posing a clinical challenge. Clinical practitioners and laboratory personnel should be aware of the epidemiological, bacteriological, and molecular characteristics of serotype 24F ST162. We believe that our findings can help diagnose and treat IPD caused by serotype 24F ST162, a serotype expected to become problematic in the post-13 valent pneumococcal conjugate vaccine era.

Pyrolysis and steam gasification properties of mangroves.

The rising atmospheric CO2 emissions from the burning of fossil fuels remains a global concern. Mangrove forests, which are important for global biodiversity, are known for their high carbon fixation capacity. However, the characteristics of the pyrolysis and gasification of mangroves remain unclear. Thus, this study focused on mangroves' basic pyrolysis and steam gasification properties for use as a gasification fuel. Three mangrove species: Rhizophora mucronata, Bruguiera cylindrica, and Avicennia marina, were used as experimental samples. In addition, three species of land wood were used for comparison: Eucalyptus, Japanese cedar, and Japanese cypress. In addition to the raw sample, a demineralized sample was used for each sample to account for the influence of the alkali and alkaline earth metals (AAEMs) on pyrolysis and steam gasification. Thermogravimetric analysis was performed to obtain thermogravimetric curves of mangroves and land wood. A laboratory-scale instrument for pyrolysis and gasification using a batch-type horizontal electric furnace was also used at 800 °C in an inert and steam atmosphere. The char yield of raw mangroves was high and independent of the Klason lignin content, suggesting that AAEMs influence char formation during the initial pyrolysis of the mangroves. The results of pyrolysis and gasification under steam atmosphere showed that the H2 production ratio (Steam/Inert) from mangroves was 2.52-5.33, compared to 1.76-2.35 for land woods, the addition of steam significantly enhanced the steam gasification of mangroves. Mangroves contain relatively large amounts of AAEMs, which indicates their potential as a gasification feedstock.

Trend of sociodemographic and economic inequalities in the use of maternal health services in Lao People's Democratic Republic from 2006 to 2017: MICS data analysis.

BACKGROUND: Maternal mortalities remain high in the Lao People's Democratic Republic (Lao PDR). Since 2012, to improve access to maternal health services for all women, the country implemented several policies and strategies including user fee removal interventions for childbirth-related care. However, it remains unclear whether inequalities in access to services have reduced in the post-2012 period compared to pre-2012. Our study compared the change in sociodemographic and economic inequalities in access to maternal health services between 2006 to 2011-12 and 2011-12 to 2017. METHODS: We used the three most recent Lao Social Indicator Survey datasets conducted in 2006, 2011-12, and 2017 for this analysis. We assessed wealth, area of residence, ethnicity, educational attainment, and women's age-related inequalities in the use of at least one antenatal care (ANC) visit with skilled personnel, institutional delivery, and at least one facility-based postnatal care (PNC) visit by mothers. The magnitude of inequalities was measured using concentration curves, concentration indices (CIX), and equiplots. RESULTS: The coverage of at least one ANC with skilled personnel increased the most between 2012 and 2017, by 37.1% in Hmong minority ethnic group women, 36.1% in women living in rural areas, 31.1%, and 28.4 in the poorest and poor, respectively. In the same period, institutional deliveries increased the most among women in the middle quintiles by 32.8%, the poor by 29.3%, and Hmong women by 30.2%. The most significant reduction in inequalities was related to area of residence between 2006 and 2012 while it was based on wealth quintiles in the period 2011-12 to 2017. Finally, in 2017, wealth-related inequalities in institutional delivery remained high, with a CIX of 0.193 which was the highest of all CIX values. CONCLUSION: There was a significant decline in inequalities based on the area of residence in the use of maternal health services between 2006 and 2011-12 while between 2011-12 and 2017, the largest decrease was based on wealth quintiles. Policies and strategies implemented since 2011-12 might have been successful in improving access to maternal health services in Lao PDR. Meanwhile, more attention should be given to improving the uptake of facility-based PNC visits.

The oral disposition index calculated from a meal tolerance test is a crucial indicator for evaluating differential normalization of postprandial glucose and triglyceride excursions in morbidly obese patients after laparoscopic sleeve gastrectomy.

To determine the normalization of postprandial blood glucose (PG) and triglyceride (TG) excursions in 30 morbidly obese patients with or without diabetes mellitus (DM) 1-year after they underwent a laparoscopic sleeve gastrectomy (LSG) vs. their pre-surgery data, we administered the 75-g oral glucose tolerance test (OGTT) and a meal tolerance test (MTT) using a 75-g glucose-equivalent carbohydrate- and fat-containing meal. The results were as follows; (i) Postoperative body-weight reduction was associated with DM remission and reduced multiple cardiometabolic risks. (ii) OGTT data showing postprandial hyper-insulinemic hypoglycemia in many post-surgery patients were associated with overdiagnosis of improved glucose tolerance. However, postoperative MTT data without hypoglycemia showed no improvement in the glucose tolerance vs. pre-surgery data. (iii) The disposition index (DI) i.e., [Matsuda index] × (Glucose-induced insulin secretion) was progressively worsened from normal glucose tolerance to DM patients after LSG. These post-surgery DI values measured by the MTT were correlated with 2h-plasma glucose levels and were not normalized in DM patients. (iv) The baseline, 2h-TG, and an increase in 2h-TG values above baseline were correlated with the insulin resistance index, DI, or HbA1c; These TG values were normalized post-LSG. In conclusion, the glucose tolerance curve measured by the MTT was not normalized in T2DM patients, which was associated with impaired normalization of the DI values in those patients 1-year after the LSG. However, the baseline TG and a fat-induced 2h-TG values were normalized postoperatively. The MTT can be used to assess normalization in postprandial glucose and TG excursions after LSG.

Reanalysis of Chromosomal Microarray Data Using a Smaller Copy Number Variant Call Threshold Identifies Four Cases with Heterozygous Multiexon Deletions of ARID1B, EHMT1, and FOXP1 Genes.

Introduction: Chromosomal microarray (CMA) is a highly accurate and established method for detecting copy number variations (CNVs) in clinical genetic testing. CNVs are important etiological factors for disorders such as intellectual disability, developmental delay, and multiple congenital anomalies. Recently developed analytical methods have facilitated the identification of smaller CNVs. Therefore, reanalyzing CMA data using a smaller CNV calling threshold may yield useful information. However, this method was left to the discretion of each institution. Methods: We reanalyzed the CMA data of 131 patients using a smaller CNV call threshold: 50 kb 50 probes for gain and 25 kb 25 probes for loss. We interpreted the reanalyzed CNVs based on the most recently available information. In the reanalysis, we filtered the data using the Clinical Genome Resource dosage sensitivity gene list as an index to quickly and efficiently check morbid genes. Results: The number of copy number loss was approximately 20 times greater, and copy number gain was approximately three times greater compared to those in the previous analysis. We detected new likely pathogenic CNVs in four participants: a 236.5 kb loss within ARID1B, a 50.6 kb loss including EHMT1, a 46.5 kb loss including EHMT1, and an 89.1 kb loss within the FOXP1 gene. Conclusion: The method employed in this study is simple and effective for CMA data reanalysis using a smaller CNV call threshold. Thus, this method is efficient for both ongoing and repeated analyses. This study may stimulate further discussion of reanalysis methodology in clinical laboratories.

Health of migrants and other vulnerable populations across Asian countries: Build Forward Better beyond the COVID-19 pandemic.

Global migration has been increasing since before the COVID-19 pandemic. The pandemic has clearly shown a lack of preparedness for the next public health emergency when it comes to vulnerable populations including migrants. To include the issues of migration and health in the current global health agenda, it is important to establish/strengthen a network for collaboration among various stakeholders from both the migrant-sending and host countries of migrants especially in the Asian-Pacific region. As the initial step for networking in Asia, in March 2023, a hybrid style international symposium was held in Japan and agreed on a goal and five pillars: surveillance and monitoring, risk communications, community engagement, access to health and social protection services, and supportive environments. Considering the transition of context from the COVID-19 crisis to 'Build Forward Better', through the Asian network, we will envisage the better world, where vulnerable populations including migrants will not be left behind from health security.

Reproducible Preparation of Primary Rat Hepatocyte Sheets Using a Thermoresponsive Culture Dish.

Hepatocyte transplantation has been utilized as a therapy for congenital metabolic liver diseases such as hemophilia and for liver function support in acute liver failure. Hepatocyte sheet technology using a thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-grafted dish is expected to provide an efficient cell transplantation method because the resulting hepatocyte sheet possesses extracellular matrix (ECM) on the basal surface, which enhances attachment to the target sites. However, the cultured hepatocytes consume large amounts of oxygen, leading to the loss of a few hepatocytes within the confluent culture sheet owing to a lack of oxygen. To circumvent this problem, this work demonstrates the shortening of diffusion distance, that is, the medium depth, to accelerate oxygen supply from the gas phase/medium interface to the cultured hepatocytes, allowing them to form a monolayer hepatocyte sheet. Incubation of hepatocytes with medium at a depth of 1.3 mm facilitates confluent culture of hepatocytes for 72 h, whereas viable hepatocytes decreased at 2.6 mm depth. Hepatocyte sheets are formed on a 0.5 µg/cm2 fibronectin-physisorbed PIPAAm-grafted dish during 72 h incubation at 37°C. Detachment of the cultured hepatocyte sheet from the PIPAAm-grafted dish where the surface becomes hydrophilic at 20°C is accomplished by scraping the periphery of the sheet using a cell scraper. Furthermore, the apical side of the hepatocyte sheet can be physically grabbed using a gelatin-coated membrane, and the sheet with ECM on the basal surface can be readily transferred to the target site after melting the coated gelatin at 37°C. Both methods are beneficial for creating tissue models by layering with another type of cell sheets, and for quick transplantation, such as into the subcutaneous space and orthotopic transplantation on the surface of the liver.

Efficacy of KPR-5714, a selective transient receptor potential melastatin 8 antagonist, on chronic psychological stress-induced bladder overactivity in male rats.

AIMS: Chronic psychological stress aggravates lower urinary tract symptoms. Among others, water avoidance stress is a chronic psychological stressor that plays a causal role in the exacerbation and development of bladder dysfunction in rats. In this report, the effects of KPR-5714, which is a selective transient receptor potential melastatin 8 (TRPM8) antagonist, on bladder overactivity induced by water avoidance stress were examined. METHODS: Male rats were subjected to water avoidance stress for 2 h per day for 10 consecutive days. The effects of water avoidance stress on voiding behavior using metabolic cages and histological bladder changes were investigated in rats. The involvement of bladder C-fiber afferent on voiding frequency in rats exposed to water avoidance stress was assessed using capsaicin. The effects of KPR-5714 on storage dysfunction in rats subjected to water avoidance stress were examined. RESULTS: In voiding behavior measurements, water avoidance stress-induced storage dysfunction, causing a decrease in the mean voided volume and increasing voiding frequency. A comparison of bladders from normal rats and rats exposed to water avoidance stress showed no histological differences. Water avoidance stress-induced bladder overactivity was completely inhibited by pretreatment with capsaicin. KPR-5714 showed a tendency to increase the mean voided volume and significantly decreased the voiding frequency without affecting the total voided volume in these rats. CONCLUSION: The results suggest that KPR-5714 is a promising option for treating chronic psychological stress-induced bladder overactivity.

Impact of health systems reform on COVID-19 control in Sierra Leone: a case study.

BACKGROUND: There are various impacts of COVID-19 on health systems of the world. The health systems of low- and middle-income countries are less developed. Therefore, they have greater tendencies to experience challenges and vulnerabilities in COVID-19 control compared to high-income countries. It is important to contain the spread of the virus, and likewise strengthen the capacity of health systems in order for the response to be effective and swift. The experience from 2014 to 2016 Ebola outbreak in Sierra Leone served as preparation for COVID-19 outbreak. The aim of this study is to determine how control of COVID-19 outbreak in Sierra Leone was enhanced by the lessons learned from 2014 to 2016 Ebola outbreak, and health systems reform. METHODS: We used data from a qualitative case study conducted in four districts in Sierra Leone through key informant interviews, focus group discussions, document, and archive record reviews. A total of 32 key informant interviews and 14 focus group discussions were conducted. A thematic analysis was used to analyze the data, and all transcripts were coded and analyzed with the aid of ATLAS.ti 9 software program. RESULTS: The six themes obtained were composed of categories that connect with each other and with codes to form networks. The analysis of the responses demonstrated that "Multisectoral Leadership and Cooperation", "Government Collaboration among International Partners", and "Awareness in the Community" were among the key interventions used during the control of 2014-2016 Ebola virus disease outbreak, which were applied in the control of COVID-19. An infectious disease outbreak control model was proposed based on the results obtained from the analysis of the lessons learned during the Ebola virus disease outbreak, and health systems reform. CONCLUSIONS: "Multisectoral Leadership and Cooperation", "Government Collaboration among International Partners" and "Awareness in the Community" are key strategies that enhanced the control of the COVID-19 outbreak in Sierra Leone. It is recommended that they are implemented in controlling COVID-19 pandemic or any other infectious disease outbreak. The proposed model can be used in controlling infectious disease outbreaks, especially in low- and middle-income countries. Further research is needed to validate the usefulness of these interventions in overcoming an infectious disease outbreak.