N6-methyladenosine-driven miR-143/145-KLF4 circuit orchestrates the phenotypic switch of pulmonary artery smooth muscle cells.

Pulmonary hypertension (PH) is characterized by vascular remodeling predominantly driven by a phenotypic switching in pulmonary artery smooth muscle cells (PASMCs). However, the underlying mechanisms for this phenotypic alteration remain incompletely understood. Here, we identified that RNA methyltransferase METTL3 is significantly elevated in the lungs of hypoxic PH (HPH) mice and rats, as well as in the pulmonary arteries (PAs) of HPH rats. Targeted deletion of Mettl3 in smooth muscle cells exacerbated hemodynamic consequences of hypoxia-induced PH and accelerated pulmonary vascular remodeling in vivo. Additionally, the absence of METTL3 markedly induced phenotypic switching in PASMCs in vitro. Mechanistically, METTL3 depletion attenuated m6A modification and hindered the processing of pri-miR-143/145, leading to a downregulation of miR-143-3p and miR-145-5p. Inhibition of hnRNPA2B1, an m6A mediator involved in miRNA maturation, similarly resulted in a significant reduction of miR-143-3p and miR-145-5p. We demonstrated that miR-145-5p targets Krüppel-like factor 4 (KLF4) and miR-143-3p targets fascin actin-bundling protein 1 (FSCN1) in PASMCs. The decrease of miR-145-5p subsequently induced an upregulation of KLF4, which in turn suppressed miR-143/145 transcription, establishing a positive feedback circuit between KLF4 and miR-143/145. This regulatory circuit facilitates the persistent suppression of contractile marker genes, thereby sustaining PASMC phenotypic switch. Collectively, hypoxia-induced upregulation of METTL3, along with m6A mediated regulation of miR-143/145, might serve as a protective mechanism against phenotypic switch of PASMCs. Our results highlight a potential therapeutic strategy targeting m6A modified miR-143/145-KLF4 loop in the treatment of PH.

A genetically validated approach for detecting inorganic polyphosphates in plants.

Inorganic polyphosphates (polyPs) are linear polymers of orthophosphate units linked by phosphoanhydride bonds. Polyphosphates represent important stores of phosphate and energy, and are abundant in many pro- and eukaryotic organisms. In plants, the existence of polyPs has been established using microscopy and biochemical extraction methods that are now known to produce artifacts. Here we use a polyP-specific dye and a polyP-binding domain to detect polyPs in plant and algal cells. To develop the staining protocol, we induced polyP granules in Nicotiana benthamiana and Arabidopsis cells by heterologous expression of Escherichia coli polyphosphate kinase 1 (PPK1). Over-expression of PPK1 but not of a catalytically impaired version of the enzyme leads to severe growth phenotypes, suggesting that ATP-dependent synthesis and accumulation of polyPs in the plant cytosol is toxic. We next crossed stable PPK1-expressing Arabidopsis lines with plants expressing the polyP-binding domain of E. coli exopolyphosphatase (PPX1c), which co-localized with PPK1-generated polyP granules. These granules were stained by the polyP-specific dye JC-D7 and appeared as electron-dense structures in transmission electron microscopy sections. Using the polyP staining protocol derived from these experiments, we screened for polyP stores in different organs and tissues of both mono- and dicotyledonous plants. While we could not detect polyP granules in higher plants, we could visualize the polyP-rich acidocalcisomes in the green alga Chlamydomonas reinhardtii.

TWISTED DWARF1 Mediates the Action of Auxin Transport Inhibitors on Actin Cytoskeleton Dynamics.

Plant growth and architecture is regulated by the polar distribution of the hormone auxin. Polarity and flexibility of this process is provided by constant cycling of auxin transporter vesicles along actin filaments, coordinated by a positive auxin-actin feedback loop. Both polar auxin transport and vesicle cycling are inhibited by synthetic auxin transport inhibitors, such as 1-N-naphthylphthalamic acid (NPA), counteracting the effect of auxin; however, underlying targets and mechanisms are unclear. Using NMR, we map the NPA binding surface on the Arabidopsis thaliana ABCB chaperone TWISTED DWARF1 (TWD1). We identify ACTIN7 as a relevant, although likely indirect, TWD1 interactor, and show TWD1-dependent regulation of actin filament organization and dynamics and that TWD1 is required for NPA-mediated actin cytoskeleton remodeling. The TWD1-ACTIN7 axis controls plasma membrane presence of efflux transporters, and as a consequence act7 and twd1 share developmental and physiological phenotypes indicative of defects in auxin transport. These can be phenocopied by NPA treatment or by chemical actin (de)stabilization. We provide evidence that TWD1 determines downstream locations of auxin efflux transporters by adjusting actin filament debundling and dynamizing processes and mediating NPA action on the latter. This function appears to be evolutionary conserved since TWD1 expression in budding yeast alters actin polarization and cell polarity and provides NPA sensitivity.

The cyclophilin A DIAGEOTROPICA gene affects auxin transport in both root and shoot to control lateral root formation.

Cyclophilin A is a conserved peptidyl-prolyl cis-trans isomerase (PPIase) best known as the cellular receptor of the immunosuppressant cyclosporine A. Despite significant effort, evidence of developmental functions of cyclophilin A in non-plant systems has remained obscure. Mutations in a tomato (Solanum lycopersicum) cyclophilin A ortholog, DIAGEOTROPICA (DGT), have been shown to abolish the organogenesis of lateral roots; however, a mechanistic explanation of the phenotype is lacking. Here, we show that the dgt mutant lacks auxin maxima relevant to priming and specification of lateral root founder cells. DGT is expressed in shoot and root, and localizes to both the nucleus and cytoplasm during lateral root organogenesis. Mutation of ENTIRE/IAA9, a member of the auxin-responsive Aux/IAA protein family of transcriptional repressors, partially restores the inability of dgt to initiate lateral root primordia but not the primordia outgrowth. By comparison, grafting of a wild-type scion restores the process of lateral root formation, consistent with participation of a mobile signal. Antibodies do not detect movement of the DGT protein into the dgt rootstock; however, experiments with radiolabeled auxin and an auxin-specific microelectrode demonstrate abnormal auxin fluxes. Functional studies of DGT in heterologous yeast and tobacco-leaf auxin-transport systems demonstrate that DGT negatively regulates PIN-FORMED (PIN) auxin efflux transporters by affecting their plasma membrane localization. Studies in tomato support complex effects of the dgt mutation on PIN expression level, expression domain and plasma membrane localization. Our data demonstrate that DGT regulates auxin transport in lateral root formation.

A novel putative auxin carrier family regulates intracellular auxin homeostasis in plants.

The phytohormone auxin acts as a prominent signal, providing, by its local accumulation or depletion in selected cells, a spatial and temporal reference for changes in the developmental program. The distribution of auxin depends on both auxin metabolism (biosynthesis, conjugation and degradation) and cellular auxin transport. We identified in silico a novel putative auxin transport facilitator family, called PIN-LIKES (PILS). Here we illustrate that PILS proteins are required for auxin-dependent regulation of plant growth by determining the cellular sensitivity to auxin. PILS proteins regulate intracellular auxin accumulation at the endoplasmic reticulum and thus auxin availability for nuclear auxin signalling. PILS activity affects the level of endogenous auxin indole-3-acetic acid (IAA), presumably via intracellular accumulation and metabolism. Our findings reveal that the transport machinery to compartmentalize auxin within the cell is of an unexpected molecular complexity and demonstrate this compartmentalization to be functionally important for a number of developmental processes.

Regulation of ABCB1/PGP1-catalysed auxin transport by linker phosphorylation.

Polar transport of the plant hormone auxin is controlled by PIN- and ABCB/PGP-efflux catalysts. PIN polarity is regulated by the AGC protein kinase, PINOID (PID), while ABCB activity was shown to be dependent on interaction with the FKBP42, TWISTED DWARF1 (TWD1). Using co-immunoprecipitation (co-IP) and shotgun LC-MS/MS analysis, we identified PID as a valid partner in the interaction with TWD1. In-vitro and yeast expression analyses indicated that PID specifically modulates ABCB1-mediated auxin efflux in an action that is dependent on its kinase activity and that is reverted by quercetin binding and thus inhibition of PID autophosphorylation. Triple ABCB1/PID/TWD1 co-transfection in tobacco revealed that PID enhances ABCB1-mediated auxin efflux but blocks ABCB1 in the presence of TWD1. Phospho-proteomic analyses identified S634 as a key residue of the regulatory ABCB1 linker and a very likely target of PID phosphorylation that determines both transporter drug binding and activity. In summary, we provide evidence that PID phosphorylation has a dual, counter-active impact on ABCB1 activity that is coordinated by TWD1-PID interaction.

Patient-centered Medical Home capability and clinical performance in HRSA-supported health centers.

OBJECTIVES: To evaluate the relationship between Patient-centered Medical Home (PCMH) model adoption in health centers (HCs) and clinical performance measures and to determine if adoption of PCMH characteristics is associated with better clinical performance. RESEARCH DESIGN: Data came from the Health Resources and Services Administration's 2009 Uniform Data System and the 2009 Commonwealth Fund National Survey of Federally Qualified Health Centers. Clinical performance measures included 2 process measures (childhood immunization and cervical cancer screening) and 2 outcome measures (hypertension control and diabetes control). Total and subscale PCMH scores were regressed on the clinical performance measures, adjusting for patient, provider, financial, and institutional characteristics. RESULTS: The findings showed different directional relationships, with some PCMH domains (care management, test/referral tracking, quality improvement, and external coordination) showing little or no effect on outcome measures of interest, 1 domain (access/communication) associated with improved outcomes, and 1 domain (patient tracking/registry) associated with worse outcomes. CONCLUSIONS: This study is among the first to examine the association between PCMH transformation and clinical performance in HCs, providing an understanding of the impact of PCMH adoption within safety-net settings. The mixed results highlight the importance of examining relationships between specific PCMH domains and specific clinical quality measures, in addition to analyzing overall PCMH scores which could yield distorted findings.

Keeping it all together: auxin-actin crosstalk in plant development.

Polar auxin transport and the action of the actin cytoskeleton are tightly interconnected, which is documented by the finding that auxin transporters reach their final destination by active movement of secretory vesicles along F-actin tracks. Moreover, auxin transporter polarity and flexibility is thought to depend on transporter cycling that requires endocytosis and exocytosis of vesicles. In this context, we have reviewed the current literature on an involvement of the actin cytoskeleton in polar auxin transport and identify known similarities and differences in its structure, function and dynamics in comparison to non-plant organisms. By describing how auxin modulates actin expression and actin organization and how actin and its stability affects auxin-transporter endocytosis and recycling, we discuss the current knowledge on regulatory auxin-actin feedback loops. We focus on known effects of auxin and of auxin transport inhibitors on the stability and organization of actin and examine the functionality of auxin and/or auxin transport inhibitor-binding proteins with respect to their suitability to integrate auxin/auxin transport inhibitor action. Finally, we indicate current difficulties in the interpretation of organ, time and concentration-dependent auxin/auxin transport inhibitor treatments and formulate simple future experimental guidelines.

FIMBRIN1 is involved in lily pollen tube growth by stabilizing the actin fringe.

An actin fringe structure in the subapex plays an important role in pollen tube tip growth. However, the precise mechanism by which the actin fringe is generated and maintained remains largely unknown. Here, we cloned a 2606-bp full-length cDNA encoding a deduced 77-kD fimbrin-like protein from lily (Lilium longiflorum), named FIMBRIN1 (FIM1). Ll-FIM1 was preferentially expressed in pollen and concentrated at actin fringe in the subapical region, as well as in longitudinal actin-filament bundles in the shank of pollen tubes. Microinjection of Ll-FIM1 antibody into lily pollen tubes inhibited tip growth and disrupted the actin fringe. Furthermore, we verified the function of Ll-FIM1 in the fim5 mutant of its closest relative, Arabidopsis thaliana. Pollen tubes of fim5 mutants grew with a larger diameter in early stages but could recover into normal forms in later stages, despite significantly slower growth rates. The actin fringe of the fim5 mutants, however, was impaired during both early and late stages. Impressively, stable expression of fim5pro:GFP:Ll-FIM1 rescued the actin fringe and the growth rate of Arabidopsis fim5 pollen tubes. In vitro biochemical analysis showed that Ll-FIM1 could bundle actin filaments. Thus, our study has identified a fimbrin that may stabilize the actin fringe by cross-linking actin filaments into bundles, which is important for proper tip growth of lily pollen tubes.

Chronic conditions and medical expenditures among non-institutionalized adults in the United States.

INTRODUCTION: This study sought to examine medical expenditures among non-institutionalized adults in the United States with one or more chronic conditions. METHOD: Using data from the 2010 Medical Expenditure Panel Survey (MEPS) Household Component (HC), we explored total and out-of-pocket medical, hospital, physician office, and prescription drug expenditures for non-institutionalized adults 18 and older with and without chronic conditions. We examined relationships between expenditure differences and predisposing, enabling, and need factors using recent, nationally representative data. RESULTS: Individuals with chronic conditions experienced higher total spending than those with no chronic conditions, even after controlling for confounding factors. This relationship persisted with age. Out-of-pocket spending trends mirrored total expenditure trends across health care categories. Additional population characteristics that were associated with high health care expenditures were race/ethnicity, marital status, insurance status, and education. CONCLUSIONS: The high costs associated with having one or more chronic conditions indicates a need for more robust interventions to target population groups who are most at risk.

Netographic narratives of user-generated travelogues on tourist destination image of Thailand.

The image of a tourist destination is considered a vital aspect of international travel experiences, yet research in this area remains limited. Adopting a combination of netography and qualitative research methodology, this study aims to contribute to the scientific knowledge of destination image development in Thailand by analysing online travelogues to evaluate how Chinese tourists interpret the idea of destination image. To achieve this goal, 146,641 words of Chinese internet comments containing the keyword "Thailand" from four major media sources and Chinese bloggers were gathered and analysed using netography methodology. The findings showed that there was a rise in public interest, in public forums, in the destination image of Thailand among Chinese outbound tourists. The study's results may provide important fundamental theoretical insights and inspire further investigation into the issue of destination image construction.

Effects of patient-centered medical home attributes on patients' perceptions of quality in federally supported health centers.

PURPOSE: We sought to assess patients' ratings of patient-centered medical home (PCMH) attributes and overall quality of care within federally supported health centers. METHODS: Data were collected through the 2009 Health Center Patient Survey (n = 4,562), which consisted of in-person interviews and included a nationally representative sample of patients seen in health centers. Quality measures included patients' perceptions of overall quality of services, perceptions of quality of clinician advice/treatment, and likelihood of referring friends and relatives to the health center. PCMH attributes included (1) access to care getting to health center, (2) access to care during visit, (3) patient-centered communication with health care clinicians, (4) patient-centered communication with support staff, (5) self-management support for chronic conditions, (6) self-management support for behavioral risks, and (7) comprehensive preventive care. Bivariate analysis and logistic regressions were used to examine associations between patients' perceptions of PCMH attributes and patient-reported quality of care. RESULTS: Eighty-four percent of patients reported excellent/very good overall quality of services, 81% reported excellent/very good quality of clinician care, and 84% were very likely to refer friends and relatives. Higher patient ratings on the access to care and patient-centered communication attributes were associated with higher odds of patient-reported high quality of care on the 3 outcome measures. CONCLUSIONS: More than 80% of patients perceived high quality of care in health centers. PCMH attributes related to access to care and communication were associated with greater likelihood of patients reporting high-quality care.

Race/ethnicity, insurance, income and access to care: the influence of health status.

OBJECTIVES: To examine health care access disparities with regard to health status and presence of functional limitations, a common measure of disability and multimorbidity, after controlling for individual's race/ethnicity, insurance status and income in the U.S. using the latest survey data. METHODS: Using data from the 2009 Family Core component of the National Health Interview Survey (NHIS), we examined six measures of access to care in the twelve months prior to the interview. Covariates included self-perceived health status and the presence of functional limitations, race/ethnicity, insurance status, income, and other socioeconomic characteristics. Multiple logistic regressions were used to examine the associations. RESULTS: People with functional limitations or worse health status experience greater barriers to access. Insurance status was the single factor that was associated with all six measures of access. Disparities among racial/ethnic groups in most access indicators as well as income levels were insignificant after taking into account individuals' health status measures. CONCLUSIONS: Interventions to expand insurance coverage and the Patient Protection and Affordable Care Act are expected to contribute to reducing disparities in access to care. However, to further improve access to care, emphasis must be placed on those with poorer health status and functional limitations.

Pro-Poor Hydraulic Governmentality: Sustainable Water Management and Local Livelihoods in Mekong Delta.

This article considers and adds empirical nuances to the recent conceptualization of pro-poor water management. Using the concept of pro-poor hydraulic governmentality along the Vietnam-Cambodia border of ThÆ°ong PhÆ°oc commune, we argue that water management is linked to local rural livelihoods in a complex and dynamic pro-poor mechanism. While certain policies organize local populations according to cost-effectiveness ignoring local customs, the practicalities of dealing with such constraints are much more ambivalent. This article demonstrates the structural pro-poor complexity among sand excavation, riverbank landslides, water management, local livelihoods, and populace resettlement. The government's resettlement plans and the perceptions of residents of these plans are intertwined with a wider political, economic, social, and cultural significance in the context of strong institutional power in Vietnam. Limitations and future research agenda are also indicated in the discussion and conclusion section.

The perceived value of local knowledge tourism: dimension identification and scale development.

Introduction: Local knowledge tourism encompasses the rich cultural heritage, historical narratives, and traditional practices of a specific destination. Despite its significance in enhancing the tourist experience, there is a dearth of research examining the subjective perceptions and values of visitors engaging in local knowledge tourism. Consequently, there is a pressing need to explore the composition of perceived tourist values in this unique context. Methods: Due to the exploratory nature of this research, a constructivist grounded theory and content analysis are applied to analyze the data. Results: This study identifies and conceptualizes five distinct dimensions of perceived values in local knowledge tourism: functional value, emotional value, social value, cognitive value, and self-actualization value. Furthermore, an 18-item scale is developed to measure these dimensions quantitatively. Discussion: This research makes several significant contributions: (1) it expands the scope of perceived value research within the tourism domain and enhances our understanding of the tourist experience in local knowledge tourism; (2) it provides a reliable instrument for future quantitative investigations into the behavior and mindset of local knowledge tourists; and (3) it offers theoretical foundations and practical insights for destination managers seeking to develop tourism products tailored to the preferences and expectations of local knowledge tourists.

Roboethics of tourism and hospitality industry: A systematic review.

This study aims to give a comprehensive analysis of customers' acceptance and use of AI gadgets and its relevant ethical issues in the tourism and hospitality business in the era of the Internet of Things. Adopting a PRISMA methodology for Systematic Reviews and Meta-Analyses, the present research reviews how tourism and hospitality scholars have conducted research on AI technology in the field of tourism and the hospitality industry. Most of the journal articles related to AI issues published in Web of Science, ScienceDirect.com and the journal websites were considered in this review. The results of this research offer a better understanding of AI implementation with roboethics to investigate AI-related issues in the tourism and hospitality industry. In addition, it provides decision-makers in the hotel industry with practical references on service innovation, participation in the design of AI devices and AI device applications, meeting customer needs, and optimising customer experience. The theoretical implications and practical interpretations are further identified.

Self-perception evolution among university student TikTok users: evidence from China.

The effects of short movies on social media platforms are gaining worldwide popularity and are now attracting global academic attention. Employing self-perception theory and qualitative research methodology, the study examines the influence of short video applications (TikTok) on app-user engagement and evaluates the self-perceived cognitive psychological understanding of Chinese university students. The findings show that identity, attitude change, emotional perception, and civic engagement are the most influential aspects of Chinese youths' self-perceptions. Furthermore, the positive and negative correlated components influence the distribution of short video values. Such tactical use of personality construction contributes to the present psychological research of Chinese university students.

Primary care and public health activities in select US health centers: documenting successes, barriers, and lessons learned.

OBJECTIVES: We examined primary care and public health activities among federally funded health centers, to better understand their successes, the barriers encountered, and the lessons learned. METHODS: We used qualitative and quantitative methods to collect data from 9 health centers, stratified by administrative division, urban-rural location, and race/ethnicity of patients served. Descriptive data on patient and institutional characteristics came from the Uniform Data System, which collects data from all health centers annually. We administered questionnaires and conducted phone interviews with key informants. RESULTS: Health centers performed well on primary care coordination and community orientation scales and reported conducting many essential public health activities. We identified specific needs for integrating primary care and public health: (1) more funding for collaborations and for addressing the social determinants of health, (2) strong leadership to champion collaborations, (3) trust building among partners, with shared missions and clear expectations of responsibilities, and (4) alignment and standardization of data collection, analysis, and exchange. CONCLUSIONS: Lessons learned from health centers should inform strategies to better integrate public health with primary care.

[The superiorities of applying comprehensive medical model in treating stroke in Chinese medicine hospitals].

OBJECTIVE: To study the clinical effectiveness of applying comprehensive medical model in treating stroke in Chinese medicine (CM) hospitals. METHODS: Totally 236 stroke inpatients were randomly assigned to the comprehensive treatment group (as the experimental group, 121 cases) and the conventional treatment group (as the control group, 115 cases). A standard comprehensive treatment mode was applied in the experimental group. The Chinese materia medica (CMM) or acupuncture was combined during the rehabilitation treatment. Traditional basic drug treatment method was applied in the conventional treatment group. The health education for patients and their families was launched non-systematically. The mortality, the occurrence rate of complications, short-term clinical effectiveness, as well as NIHSS, Barthel index (BI), and Oxford Handicap Scale (OHS) were compared between the two groups. RESULTS: Before treatment there was no statistical difference in NIHSS score, BI, or OHS score between the two groups (P > 0.05). On day 21 and 90 of the treatment, there was statistical difference in NIHSS score, BI, and OHS score (P < 0.01). Better results were obtained in the experimental group. On day 21 and 90 of the treatment, the total effectiveness rate was 80.99% and 88.43% in the experimental group, higher than that of the control group (61.74% and 72.17% respectively), showing statistical difference (P < 0.05, P < 0.01). The occurrence rate of common complications in the experimental group was obviously lower than that in the control group (30.58% vs 69.56%, P < 0.01). There was no statistical difference in the mortality between the two groups (P > 0.05). CONCLUSIONS: The application of the comprehensive medical model in treating stroke in CM showed better effects than using traditional basic drug treatment method. Treatment of stroke by CM showed superiorities.

Internationalization, cultural appreciation and institutional governmentality for quality control in transnational higher education cooperation: An empirical assessment.

This article examines the dynamic mechanism of cultural appreciation and institutional governmentality to ensure successful quality control in a transnational higher education collaboration context. Adopting participatory action research and a case study approach, this paper investigates the quality control system in a Chinese tourism university. The present study finds that mutual cultural appreciation, responsible government guidance and institutional governmentality are essential quality control measures for transnational higher education cooperation. The quality control system is suggested to be established to enrich and improve the quality standards of joint international higher education collaboration. This study proposes to expand the international influence and recognition of China-foreign education collaboration through quality international exchange and cooperation.