Japan Tobacco (Japan)

Alveolar regeneration requires coordinated alveolar type 1 (AT1)/alveolar type 2 (AT2) responses and an intact epithelial barrier. In vitro systems, however, rarely couple lineage control with high resistance. Here we establish a concise workflow in which human AT2-like progenitors are expanded in a medium containing a small-molecule cocktail (Y-27632/A-83-01/CHIR99021; YAC), followed by air-liquid interface (ALI) culture in medium without the YAC cocktail (hereafter referred to as the YAC(-) condition). Compared with submerged culture, ALI increased AT1(AQP5, AGER, and PDPN) and AT2 (SFTPB, SFTPC, and SFTPD) markers. Within ALI, β-catenin inhibition by XAV939 elevated AT1 features while retaining AT2 protein expression, but was accompanied by reduced AT2 marker expression at the transcriptional level compared with the YAC(-) condition. Transepithelial electrical resistance entered the kΩ range by ALI day 10 and remained high across ALI conditions, with hematoxylin and eosin staining confirming a continuous single-layer epithelium. Passage reduced absolute marker levels and resistance but preserved the rank order across conditions. In scratch assays, ALI monolayers closed wounds over 24-48 h, and spatial immunofluorescence staining showed surfactant protein-C enrichment at the leading edge and podoplanin predominantly behind the front. Collectively, these findings demonstrate that β-catenin modulation within ALI biases the alveolar epithelial profile toward AT1-like features, whereas the YAC(-) condition sustains a balanced AT1/AT2 state with superior epithelial barrier performance and repair capacity. This platform provides a single-monolayer system suitable for dissecting alveolar epithelial differentiation and functional maintenance.

Community-based health insurance (CBHI) is implemented in Ethiopia to enhance healthcare access and provide financial risk protection for the vulnerable people. However, it falls short in fulfilling customers' needs and could be challenged by potential constraints, needing further investigation. This study was to assess factors associated with client satisfaction and implementation challenges for CBHI in Ethiopia. A mixed-methods (quantitative and qualitative) survey was employed using face-to-face interviews combined with key informant interviews (KII) using appreciative inquiry to learn more about the practice of CBHI. The quantitative part employed structured questionnaires, and interviews were conducted among a random sample of 782 CBHI beneficiaries using the pretested and validated Ethiopian Health Insurance Agency (EHIA) tool holding multiple dimensions, to measure client satisfaction level and quality. In addition, KII was conducted with 16 key informants from program implementer and facility planning units. The data were analyzed and presented in descriptive statistics. The KII were transcribed, translated, and analyzed using descriptive narrative analysis. A total of 782 CBHI beneficiaries with mean age of 47.6 (±11.6) years and 60.4% females were interviewed. With regard to dimensions of satisfaction, 74.3% reported that CBHI supported services at health facilities are accessible while 67.1% have trust in the health system and 61% reported to get quality services. Almost all were satisfied with the opening hours for service and information provided about the CBHI program. However, some respondents express dissatisfaction with the collection process and registration fee. Overall, about 32.4% (95% CI: 29.1-35.8) of the beneficiaries were satisfied with the CBHI services. CBHI implementation is at full scale in the area but its implementations are challenged by limited resource availability, administrative processes, insurance coverage, availability of essential services, and payments schemes. On the other hand, age (AOR = 3.66; 1.36-9.87), family size (AOR = 4.02; 2.13-7.59), education (AOR = 2.21; 1.34-3.66), and illness history (AOR = 3.15; 1.94-5.12) were associated with dissatisfaction with CBHI services. Only one-third of CBHI users were found to be satisfied with the service and it is associated with age, literacy, family size, and illness history. Key challenges cited include drug shortages, limited laboratory services, and bureaucratic policies, which should be addressed well.

Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by the accumulation of fat in the liver in the absence of excessive alcohol consumption or a secondary cause of hepatic steatosis. The prevalence of NAFLD is increasing worldwide and its management has become a public health concern. Animal models are traditionally used to elucidate disease mechanisms and identify potential drug targets; however, their translational aspects in human diseases have not been fully established. This study aimed to clarify the utility of animal models for translational research by assessing their relevance to human diseases using gene expression analysis. Weighted gene co-expression network analysis of liver tissues from Western diet (WD)-induced NAFLD mice was performed to identify the modules associated with disease progression. Moreover, the similarity of the gene co-expression network across species was evaluated using module preservation analysis. Nineteen disease-associated modules were identified. The brown module was positively associated with disease severity, and functional analyses indicated that it may be involved in inflammatory responses in immune cells. Moreover, the gene co-expression network of the brown module was highly preserved in human NAFLD liver gene expression datasets. These results indicate that WD-induced NAFLD mice have similar gene co-expression networks (especially genes associated with inflammatory responses) to humans and are thought to be a useful experimental tool for preclinical research on NAFLD.

The tissue culture process is usually involved in gene transfer and genome editing in plants. Like other species, there is enormous variation among wheat genotypes in tissue culture response. In the rapidly advancing system of CRISPR/Cas9 for genome editing, particle bombardment has received increasing attention as a delivery method for a large amount of nucleic acids and RNA-protein complexes. However, the efficiency of transformation by particle bombardment has been low in wheat, and only a limited number of varieties have been transformed. In this study, replacement of maltose with trehalose as an osmolyte for high osmotic treatment for the protection of tissues from physical impacts improved callus formation in immature wheat embryos and efficiency of transformation and genome editing in varieties that are relatively poor in tissue culture response. The range of varieties amenable to biolistic transformation and genome editing may be expanded by this modification.

Tongkat Ali (TA), also known as Eurycoma longifolia, has been used as a traditional herbal medicine for anti-aging, evidenced by clinical trials presenting the beneficial effects on energy, fatigue, and mood disturbance. We have recently shown that TA supplementation dose-dependently enhances the rest-activity pattern in C57BL/6 mice. Since destabilization of wakefulness and sleep is one of the typical symptoms of not only the elderly but also narcolepsy, we performed sleep analysis with and without dietary TA extract supplementation in middle-aged (10-12 months old) wild-type (WT) and narcoleptic DTA mice. We found that TA supplementation enhanced diurnal rhythms of locomotion and temperature in a time-of-day-dependent manner in WT mice but attenuated in DTA mice. In WT mice, TA supplementation consolidated wakefulness with a long bout duration and led to less entries into the sleep state during the active period, while it consolidated NREM sleep with long bout duration during the resting period. Neither disturbed sleep and wake cycles nor cataplexy was sufficiently improved in DTA mice. EEG spectral analysis revealed that TA supplementation enhanced slow wave activity (SWA) at both delta and low delta frequencies (0.5-4.0 and 0.5-2.0 Hz) during the light period, suggesting TA extract may induce vigilance during the active period, which then elicits a rebound effect during the resting period. Interestingly, DTA mice also slightly, but significantly, increased SWA at low frequencies during the light period. Taken together, our results suggest that TA supplementation enhances the Yin-Yang balance of sleep, temperature, and locomotion in WT mice, while its efficacy is limited in narcoleptic mice.

Nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH) is caused by various factors, including genetic and/or environmental factors, and has complicated pathophysiological features during the development of the disease. NAFLD/NASH is recognized as an unmet medical need, and NAFLD/NASH animal models are essential tools for developing new therapies, including potential drugs and biomarkers. In this review, we describe the pathological features of the NAFLD/NASH rat models, focusing on the histopathology of hepatic fibrosis. NAFLD/NASH rat models are divided into three categories: diet-induced, genetic, and combined models based on diet, chemicals, and genetics. Rat models of NASH with hepatic fibrosis are especially expected to contribute to the development of new therapies, such as drugs and biomarkers.