Circadian rhythm disruption is increasingly recognized as a contributor to chronic inflammatory disorders; however, its specific significance and underlying mechanisms in chronic obstructive pulmonary disease (COPD) remain unclear. This study aimed to identify circadian rhythm-associated biomarkers in COPD and explore their diagnostic value, immune correlations, and therapeutic potential. This study integrated four lung transcriptomic datasets from the public GEO database (GSE151052, GSE38974, and GSE76925 as the discovery set, and GSE47460 as the validation set). Differentially expressed circadian rhythm‑related genes (DECRRGs) were identified by intersecting differentially expressed genes with circadian rhythm‑related genes. Functional enrichment analyses (GO and KEGG) were performed, and three machine learning algorithms were applied to screen for signature DECRRGs. An exploratory risk stratification model based on multivariate logistic regression was constructed and evaluated. Immune cell infiltration was assessed using CIBERSORT, and single-cell RNA sequencing analysis was conducted to localize the distribution of key circadian rhythm genes within specific lung cell populations. Finally, the expression of a core gene CHRNA1 was validated by qRT-PCR in peripheral blood samples from COPD patients and healthy controls. We identified eight circadian rhythm-associated feature genes, among which CHRNA1 emerged as a consistently upregulated hub gene in COPD. An exploratory risk stratification model based on these genes exhibited good discriminatory ability in the discovery cohort (AUC = 0.856, 95% CI: 0.806-0.902). Differential expression of CHRNA1 was validated in an independent cohort and correlated significantly with pro-inflammatory immune infiltration, including increased M1 macrophages and CD8 ⁺ T cells. Single-cell transcriptomics further localized CHRNA1 expression predominantly within B cells in COPD lung tissue. In silico drug screening and ceRNA network analysis predicted potential therapeutics (e.g., amitriptyline, rocuronium bromide) and regulatory miRNAs/lncRNAs. Finally, qRT-PCR confirmed a marked upregulation of CHRNA1 in peripheral blood from COPD patients (*p* < 0.0001). Our findings suggest that CHRNA1 may serve as a candidate circadian rhythm‑associated immunomodulator in COPD. It demonstrates consistent upregulation across cohorts and shows a significant association with pro‑inflammatory immune infiltration. Single-cell analysis revealed that CHRNA1 is predominantly expressed in pulmonary B cells. The exploratory risk stratification model and predicted therapeutic candidates highlight the translational potential of targeting circadian disruption in COPD, though prospective validation is needed before clinical application.
Physical inactivity increases with age, yet evidence on physical activity (PA) among older adults affected by forced displacement remains scarce, particularly in refugee camp settings. We aimed to assess PA levels and identify associated factors among Rohingya older adults aged ≥60 years living in refugee camps in Southern Bangladesh. A cross-sectional survey was conducted between May and August 2024. PA was assessed using the Physical Activity Vital Sign (PAVS), alongside socio-demographic, health, and lifestyle variables collected via a pre-tested semi-structured questionnaire. in line with WHO guidelines, participants were classified as active (≥150 minutes of moderate to vigorous PA per week) or low active (<150 minutes of moderate to vigorous PA per week). Multivariable Firth penalised logistic regression was used to examine factors associated with meeting PA guidelines. The sample comprised 943 participants (mean age 67.5 years), of whom 70% were aged 60-69 years and 66% were male. Overall, 95% were classified as low active. In adjusted analyses, only older age (≥70 years) was significantly associated with lower odds of meeting PA guidelines (OR 0.31, 95% CI 0.11-0.70). Fewer than one in twenty Rohingya older adults met recommended PA levels. These findings highlight the need for tailored, culturally appropriate PA interventions, with particular attention to adults aged 70 years and older.
Although bar charts are widely used in scientific publications, their rasterized format within Portable Document Format (PDF) files complicates automated data extraction, hindering large-scale evidence synthesis and meta-analysis. To address this, we developed and evaluated an automated pipeline for extracting quantitative data from bar charts embedded in the biomedical literature. The four-stage pipeline comprises (1) image extraction and panel segmentation, (2) optical character recognition (OCR)-based text detection, (3) image disassembly to identify chart components, and (4) data reconstruction using numeric parsing and axis-based interpolation. The system combines edge detection, morphological operations, and convolutional neural network (CNN)-based figure classification using a transfer-learned Inception v3 model. Performance was validated on randomized controlled trials in age-related macular degeneration, with manually annotated values from a semi-automated labeling tool as the reference standard, and agreement was assessed using Bland-Altman analysis. Across 28 bar charts from ten publications, the pipeline correctly recognized 92.9% (95% confidence interval [CI], 77.4-98.0) of figure types, 96.0% (95% CI, 94.2-97.3) of text blocks, and 79.1% (95% CI, 74.7-83.0) of bars. For numerical reconstruction, 81.2% (95% CI, 76.3-85.2) of bar values fell within ±5% of the reference standard, 63.0% (95% CI, 57.3-68.3) within ±2%, and 48.6% (95% CI, 43.0-54.3) within ±1%. Bland-Altman analysis showed a small negative bias of -0.18 (95% CI, -0.34 to -0.02), with 94.9% of differences within the limits of agreement. Most outliers arose from OCR digit misclassification or ambiguous bar boundaries. This proof-of-concept study demonstrates the feasibility of automated data extraction from bar charts using a hybrid approach that combines image-processing heuristics with CNN-based classification. Although currently limited to bar charts and a single clinical domain, the pipeline represents a step toward scalable, end-to-end systems for automated evidence extraction to support meta-analyses across the biomedical literature.
Pharmacists are expanding their participation in veterinary healthcare teams and assuming roles beyond traditional dispensing duties. However, the scope of these collaborative practices and the degree of mutual recognition vary substantially across regions. This scoping review aimed to organize the existing literature on collaboration between veterinarians and pharmacists, clarifying current roles, practical applications, and professional perceptions within the veterinary field. A scoping review was conducted to examine the roles, practices, and perceptions associated with veterinarian-pharmacist collaboration in veterinary medicine and related fields. Two researchers independently searched PubMed, Web of Science, the Cochrane Library, and Ichushi-Web, and additionally screened Google Scholar to identify gray literature (from database inception to August 2025, in English or Japanese). Records were independently screened at the title/abstract and full-text levels using predefined eligibility criteria, and relevant studies were identified. The search yielded 239 records, of which 16 studies published between 2007 and 2024 were included. Studies were conducted primarily in the United States (n = 7), New Zealand (n = 3), and Japan (n = 3); one study collected data from both Japan and Taiwan. Most studies employed cross-sectional survey designs. Pharmacist roles most frequently involved compounding and dispensing for animal patients (62.5%), followed by drug information (DI) and consultation (37.5%), inventory and supply management (25.0%), client education (18.8%), and safety and exposure control (12.5%). This scoping review demonstrates that veterinarian-pharmacist collaboration is described within a limited and regionally variable evidence base, with pharmacists most often contributing through compounding/dispensing and drug information support. Sustained and scalable implementation will require improved mutual understanding of professional roles, strengthened veterinary-specific education for pharmacists, and more robust empirical research to inform collaborative practice models.
While a growing number of studies have shown positive effects of perturbation-based balance training on balance recovery after tripping, this training has employed specialized equipment that may pose a barrier for wider adoption. To address this, the purpose of this pilot trial was to evaluate the feasibility and preliminary efficacy of a novel, low-resource (i.e., not requiring specialized equipment) version of perturbation-based balance training referred to here as task-specific step training. Thirty community-dwelling older adults (mean (SD) age: 71.8 (4.4) years) were recruited and allocated to either step training (n = 10), traditional treadmill perturbation-based balance training (n = 10), or a control group involving no training (n = 10). Participants were then exposed to two overground laboratory-induced trips while walking on a walkway. Results showed the step training group exhibited an initial recovery step that was 9.0% body height longer (p < 0.001) and 0.25 m/s faster (p = 0.011) than the control group.The step training group also exhibited a 4.8% body height longer recovery step, and a fall rate that was 39% lower (p = 0.037) when compared to the treadmill training group after lab-induced trips. While promising, these results should be interpreted with caution give the modest sample size and a potential bias between groups with respect to safety harness usage during laboratory-induced trips. A future trial with adequate statistical power to better evaluate efficacy and effectiveness of this step training on real-world trip and fall risk appears warranted. The study was registered on clinicaltrials.gov (NCT05734443).
Surface plasmon resonance (SPR) enables label-free detection of binding kinetics and has been widely applied to the biophysical characterization of molecular interactions such as antibody-antigen binding. With the advent of high-throughput SPR (HT-SPR) instruments, hundreds of binding interactions can be detected simultaneously, combining the details of kinetic measurements with the capability of large-panel biomolecule screening. However, binding kinetics analysis for large panels of antibody or antigen often requires a combination of fitting strategies to address different types of sensorgrams. While software packages exist for SPR binding kinetics data analysis, they are associated with a number of limitations: 1) currently most of the software packages are proprietary, prohibiting widespread use; 2) most of the software packages, including open source packages, are designed primarily for low-throughput data analysis, making analyzing a large number of kinetics data sets labor-intensive; 3) the software typically requires multiple iterative user-interface interactions when analyzing large data sets. Here, we present htrSPRanalysis, an open source R package designed primarily for high-throughput binding kinetics data analysis, currently focusing on 1:1 binding analysis. htrSPRanalysis leverages the increasingly commonplace multi-core computing architecture to efficiently analyze a large number of sensorgrams with minimal user-interface interaction. It also offers automated generation of analysis output for all sensorgrams. Furthermore, beyond manual optimization of sensorgram fitting strategies, htrSPR analysis accelerates the analysis process by providing automated procedures to determine the optimal concentration range, choose the optimal dissociation window for fitting, and detect bulk shift. The high-throughput functionalities and automation of fitting optimization makes htrSPRanalysis especially useful for speeding up data analysis to get results for implementing further steps in therapeutic antibody discovery research.
Primary care physicians (PCPs) are on the front lines of health promotion and disease prevention, yet evidence supports a longstanding gap in physicians' capacity to counsel about diet, a key factor in patient health and chronic disease risk. Momentum to address the nutrition training needs of physicians is growing; however, challenges PCPs experience due to inadequate training are further complicated by clinical practice barriers. This study seeks to better understand the complex problem PCPs face addressing nutrition with patients to inform potential solutions. This qualitative study takes a design thinking approach to understanding the problem by learning from PCPs about their experience, challenges, and priorities addressing nutrition with patients. Findings from semi-structured interviews with 32 PCPs revealed that while they value nutrition and believe they have an important role in addressing diet with patients, their role as a generalist in addressing nutrition lacks clarity. PCPs describe learning their current approach to nutrition primarily outside of formal medical training, resulting in variability and inconsistency in approaches and recommendations and general frustrations about their ability to give actionable guidance. While registered dietitians (RDs) are often viewed as valuable support for patients, PCPs describe multiple challenges to RD utilization and subsequently at times may be the only ones with the opportunity to address diet with patients. Additional system- and societal-level barriers, such as time limitations and the food environment, further complicate PCPs' experiences advising patients about nutrition. Main limitations of this study are limited generalizability from recruiting graduates of a single medical school and PCPs with greater interest in nutrition or more frustration with their current approach to nutrition may have been more likely to participate. Our results support the need to clearly define a realistic role for PCPs as generalists addressing nutrition. Clarified expectations in turn can inform nutrition training of physicians and guiding frameworks and strategies for giving practical, evidence-based nutrition guidance. PCPs also need increased support through access to resources, strategies to improve RD collaboration, and attention to system- and societal-level challenges influencing dietary interventions in primary care.
High temperature is one of the major environmental stressors that severely affects plant growth. Paris polyphylla var. yunnanensis, a traditional Chinese herbal medicine, is sensitive to high temperature. However, the underlying mechanisms of its response to high temperature remain unclear. In this study, we investigated the physiological and proteomic change of P. polyphylla var. yunnanensis under different treatments (25°C, 30°C, 35°C, 40°C). Our results showed that high temperature directly impaired photosynthesis and disrupted metabolism, evidenced by reduced chlorophyll and photosynthetic rate, as well as accumulated proline and increased conductivity. A total of 893 differentially expressed proteins (DEPs) were identified, with significant changes in the expression levels of enzymes associated with protein processing and synthesis. Additionally, the expression levels of key proteins involved in the circadian pathway and the glutathione pathway were also notably upregulated. Dynamic changes in the endocytosis and autophagy-related proteins ATG3 and ATG8C were also observed, suggesting that these processes may play a significant protective role under high-temperature stress. Overall, this study provides an important starting point for improving the heat tolerance of P.polyphylla var. yunnanensis through genetic engineering.
The coupled function of physiology and behavior is crucial for generating survival responses. Generally, the cardiovascular system undergoes rapid adjustments during scape behavior to increase oxygen delivery to muscle tissues. In contrast, passive antipredator responses such as retraction in snails may impose mechanical constraints on the circulatory system. Here, we evaluated the cardiovascular response underlying the scape response (through induced retraction) and the physical activity in the invasive land snail Cornu aspersum. We quantified heart rate and heart rate variability in adult snails using laser optocardiography under the two conditions. We found that heart rate increased from retracted to the moving state and was accompanied by changes in heart rate variability and reduction of cardiac irregularities. Also, we found that locomotion intensity and body size did not influence these cardiac parameters. Our results suggest that metabolic demands, neural regulation and mechanical configuration collectively alter heart rate and heart rate variability. Thus, cardiovascular function is strongly dependent on behaviorally induced mechanical shifts in the circulatory system and on a "fight-or-flight"-like response.
Venom-induced compartment syndrome (VICS) is a rare but severe complication of snakebite which poses unique challenges in diagnosis and treatment. Published literature largely focuses on VICS in the North American context, while information on clinical presentations, diagnostic and treatment approaches, clinical outcomes and associated challenges from the world's most snakebite endemic regions remain fragmented and scarce. A scoping review using PRIMSA-ScR methodology of the global literature on VICS was performed by searching the PubMed, Embase, and Cochrane databases. Literature was divided into 1) Case reports on VICS and 2) General literature. Clinical data from eligible case reports was extracted to describe management of VICS and associated challenges. Available evidence on diagnostic and treatment strategies was extracted from the general literature. Of 115 cases of VICS analyzed, most were from Europe (40%); Only 13% and 6% were reported from South-East Asia and Africa respectively. Viperids caused 73% of bites and upper extremities were most frequently affected (63%). Compartment pressure was measured in 38% of patients. Compartment pressure was more commonly measured in the 12% of non-surgically treated patients, none of whom developed ischaemic contracture or required amputation. Coagulopathy was the most common systemic toxicity, present in 40% of patients at admission. Results from eight animal studies support the use of antivenom for treating VICS. Analysis of 17 human observational studies suggests an overdiagnosis of VICS using clinical symptoms alone, highlighting the need to investigate diagnostic tools, such as ultrasound, for diagnosing compartment swelling. Antivenom as first-line treatment for VICS is supported by animal and human observational studies. However, additional data is needed to inform decision-making on when fasciotomy is indicated as a rescue therapy. Improved evidence generation will depend on the collection of high-quality clinical and diagnostic observational data from patients treated with antivenom in snakebite-endemic regions.
Social support, which is an essential aspect influencing health, has resulted in the development of several approaches for assessment in cancer patients. The Illness-Specific Social Support Scale Short Version-8 (ISSS-8) effectively evaluates social support among diverse patient demographics; however, its psychometric validity has yet to be established in Asian cultural contexts. The objectives of this study were to translate and culturally adapt the ISSS-8 for the Thai setting and to assess its psychometric properties in patients with hematological malignancies (HMs). This study employed a convenience sampling method to select patients with HMs undergoing hospitalization at three tertiary institutions in Northeastern Thailand. Psychometric testing was conducted following the translation and cross-cultural adaptation of the ISSS-8 into Thai. A total of 350 patients were recruited. Participants were randomly divided into two groups for exploratory factor analysis (EFA) (n = 200) and confirmatory factor analysis (CFA) (n = 150). The EFA of the eight items yielded a loading from a two-factor model comprising Positive Support and Detrimental Interactions, which explained 82.44% of the variance. Cronbach's alpha (0.80) and item-total correlations (rho = 0.33-0.65) demonstrated acceptable reliability of the ISSS-8, while test-retest reliability was high (ICC = 0.924--0.934). The average variance extracted (AVE) demonstrated convergent validity for all ISSS-8 subscales, with AVEs ranging from 0.76 to 0.80. Moreover, the total ISSS-8 scale demonstrated a statistically significant but weak positive correlation with the Stanford Inventory of Cancer Patient Adjustment (r = 0.244, p < 0.001), while the Detrimental Interactions subscale was significantly and weak negatively correlated with it (r = 0.237, p < 0.001). These findings suggest that the ISSS-8 captures distinct psychosocial and illness-specific relational constructs that differ from coping self-efficacy and psychological adjustment measured by the SICPA. The ISSS-8 is a short, accurate, and valid instrument for measuring social support in Thai patients with HMs. As a result, healthcare professionals can use the ISSS-8 to assess social support in both research and clinical settings. The findings emphasize the necessity of integrating social support evaluation into cancer management and family-centered care, thereby supporting comprehensive and adaptive healthcare delivery in Thailand.
Typhoid is a significant global health challenge due to its high pathogenicity and antimicrobial resistance. Salmonella typhi (S.typhi) can switch its lifestyles between biofilm and planktonic phase which allows it to evade host defenses and develop resistance to antibiotics. Salmonella sp. harbors multiple genes encoding efflux-pumps systems whose up-regulation contributes to multi-drug resistance (MDR) and extensive drug-resistance (XDR). To overcome the battle against resistant S. typhi strains, novel non-antibiotics inhibitors are required for inhibitory application. This study assesses the inhibitory effect of lignans against drug resistance of S. typhi. Clinical resistant and sensitive strains of S. typhi were obtained and characterized. The inhibitory effect of lignans, specifically Schisandrin A and B, purified from the plant Schisandra chinensis, are found to be effective non-antibiotic inhibitors were evaluated through standard microbiological techniques like growth curve and time-kill assays. Impact on bacterial morphology was analyzed using scanning electron microscopy (SEM). Our study explores two approaches, such as efflux pumps (EPs) inhibition and antibiofilm assays. Using colony-forming unit (CFU) assays, growth curve analysis, and SEM imaging, we observed significant bacteriostatic effects, with Schisandrin B causing notable membrane disruption. Schisandrin B also showed remarkable biofilm inhibition (90.33%) and strong efflux pumps inhibition. This study offers a strong basis for future research on addressing antibiotic resistance in clinically relevant pathogens.
Getah virus (GETV), a mosquito-borne alphavirus, poses an emerging threat to public health with its increasingly broad host spectrum. While glycosaminoglycans (GAGs) serve as critical attachment factors for many alphaviruses and the low-density lipoprotein receptor (LDLR) facilitates the cellular entry of several members, the precise viral determinants governing these interactions and their implications for viral virulence remain poorly defined. Here, we introduced an H86Y substitution, a potential adaptive mutation site, within the E2 glycoprotein of GETV using reverse genetics. The H86Y mutant replicated more efficiently in mosquito C6/36 cells but was consistently attenuated across several mammalian cell lines. In susceptible mouse models, H86Y infection led to reduced viral loads, milder histopathology, and lower inflammatory responses compared with the parental virus, yet still elicited robust protective immunity in adult mice. Mechanistically, a series of functional assays, including infection in GAG-deficient cells, decoy inhibition, co-immunoprecipitation, receptor overexpression and knockdown, and biolayer interferometry, demonstrated that the residue 86 in E2 glycoprotein is a critical determinant for GETV binding to both GAGs and LDLR. The H86Y mutation concurrently reduces these interactions, contributing the impairment of virus attachment and entry into mammalian cells. Furthermore, the GAG-binding site functionally overlaps with the LDLR interaction interface. In LDLR-deficient suckling mice, the impaired replication of H86Y persisted in examined tissues. However, pre-treatment with heparinase nearly completely eliminated this attenuation phenotype, further confirming that LDLR and GAG are the key host factors mediating attenuation phenotype for H86Y. In summary, the residue 86 of the GETV E2 glycoprotein represents a determinant of viral virulence, and an H86Y mutation attenuates GAGs and LDLR-dependent infection, providing mechanistic insights into alphavirus-host interactions and a potential target for antiviral and vaccine development.
Bacterial vaginosis (BV) is highly prevalent in women of African origin and has been associated with adverse birth outcomes (ABO), particularly preterm birth (PTB). However, evidence from sub-Saharan Africa remains limited, and while a previous study conducted in this area examined the prevalence of BV, its impact on pregnancy outcomes was not assessed. This study assessed the prevalence and risk factors of BV in early pregnancy and examined its association with ABO in the middle belt of Ghana. This analysis was nested within a prospective cohort of pregnant women in the Kintampo North Municipality and Kintampo South District. Pregnant women <20 weeks' gestational age were screened, consented, enrolled and followed through delivery until one year after birth. At enrolment, sociodemographic, behavioural, and obstetric data were collected, along with biological samples, including vaginal swabs for Gram staining and Nugent scoring. BV status was categorized as BV-negative (0-6) or BV-positive (7-10) and ABO included PTB (<37 weeks), low birthweight (LBW < 2.5 kg), and foetal loss. Data were double-entered in REDCap and analysed in R. Associations between BV and ABO were assessed using chi-square tests, and logistic regression was used to identify risk factors for BV. Of 1,180 pregnant women enrolled, 355 (30.1%) were BV-positive. PTB (9.9% vs. 8.7%; p = 0.50), LBW (13.8% vs. 12.0%; p = 0.39), and foetal loss (4.5% vs. 2.8%; p = 0.14) were all higher among BV-positive women, but none reached statistical significance. Younger age and unmarried status were associated with BV. Unmarried women (AOR 1.63; 95% CI: 1.20-2.22) and those reporting one sexual encounter per week (AOR 1.55; 95% CI: 1.01-2.37) had increased odds of BV. BV is highly prevalent in early pregnancy in this Ghanaian cohort, particularly among younger and unmarried women, but was not significantly associated with ABO. Targeted screening among high-risk groups is necessary, and future molecular approaches may better characterise BV subtypes linked to ABO in African settings.
This study investigates how the spatial structure of Metroidvania game worlds relates to the layout of key functional nodes. Using Hollow Knight, Prince of Persia: The Lost Crown, and Blasphemous 2 as case studies, the maps were transformed into graph structures based on spatial data compiled from gameplay observation, in-game maps, community maps, and verification through gameplay videos. On the basis of this graph representation, the study adapts the Nearest Neighbor Index by replacing Euclidean distance with network shortest-path distance in order to evaluate the distribution patterns of save points, teleport points, and boss points. It further employs multi-source k-hop reachability analysis to measure service coverage and redundancy. Finally, closeness centrality and betweenness centrality are calculated on the full network, and Spearman rank correlation is used to test the association between functional areas and centrality. The results reveal clear differences in the structural roles of functional nodes across the three games. In Hollow Knight and The Lost Crown, save points are relatively dispersed, weakly coupled with centrality, and function as a distributed safety net. In Blasphemous 2, save points exhibit higher small-radius coverage and redundancy and are more strongly aligned with central positions, indicating a hub-like stronghold structure. Teleport systems likewise reflect different design strategies, ranging from exploration-oriented sparse layouts to later-stage mobility optimization. Boss points in all three games generally remain structurally isolated, showing low overlap and weak centrality association. These findings suggest that differences in the placement of save points, teleport points, and boss points reflect distinct structural design strategies, and that network analysis can provide a quantitative basis for evaluating and optimizing such layouts in level design.
Decentralization of antiretroviral therapy (ART) brought treatment to millions of people in Sub-Saharan Africa. However, the implications of such decentralization are still not fully understood. The study included 60 semi-structured interviews with community leaders, healthcare providers, patients, and community members served by a rural mission hospital in Zimbabwe, conducted in January-March 2019. Interviews were transcribed and translated into English and coded using an iterative codebook. Coded extracts were grouped into themes and categories, systematically using QDA Miner qualitative analysis software. The study reveals that treatment-for-all policies and decentralization of ART have indeed improved access to care (as experienced by the study participants). However, the study shows that these policies also had unintended negative consequences. Stigma and complex social relations significantly affect the practices and experiences of ART outreach care, for both patients and healthcare providers, and may hamper attempts to reduce stigma as well as increase physical and social barriers to accessing ART. The study's results highlight the need to understand the consequences of the decentralization of HIV care, as well as the barriers and facilitators associated with ART outreach in order to improve quality of care and provide true accessibility to HIV care for all.
Neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD), are progressive disorders with limited therapeutic options. Centella asiatica (C. asiatica), a medicinal and edible plant, has been reported to exert neuroprotective and anti-neuroinflammatory properties. Yet, the mechanisms underlying its effects against neurodegenerative diseases remain largely unclear. We employed an integrative strategy combining network pharmacology, transcriptomic analyses, machine learning and molecular docking to prioritize disease-associated molecular networks and candidate compound-target relationships in AD, PD and HD. Sixteen candidate constituents of C. asiatica met the predefined drug-likeness, gastrointestinal absorption and blood-brain barrier permeability criteria, yielding 370 unique predicted targets. Disease-gene mining identified 983 AD-associated genes, 1,103 PD-associated genes, and 3,316 HD-associated genes. Integration of compound targets, disease-associated genes, and transcriptomic profiles prioritized five hub genes in PD (CCKAR, MAPK8, PSEN2, SLC6A3, and TH), four in AD (APP, PGK1, PIK3CA, and TTR), and four in HD (CHRND, HSP90AA1, PRKCQ, and TH). Enrichment analyses highlighted disease-relevant processes involving neurotransmitter signalling, cAMP and calcium pathways, MAPK-related responses and inflammatory regulation. ROC analyses provided additional support for the discriminatory performance of the prioritized genes in independent datasets, whereas molecular docking identified favourable predicted Vina docking scores and structurally plausible interactions between selected compounds and hub targets. This integrative computational analysis prioritizes candidate C. asiatica constituents, putative disease-associated targets, and molecular pathways in AD, PD, and HD. The findings provide a foundation for subsequent biochemical, cellular, and in vivo validation.
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Animal husbandry is widely practiced on the household scale in communities in low- and middle-income countries (LMICs) and, while having economic and health benefits, exposes household members to risk of zoonotic infections to an extent that is unclear. While demand for georeferenced information on infectious disease risk factors and drivers is growing, spatial variation in livestock ownership remains poorly characterized at high resolution. This study aimed to use geostatistical methods to model and map the prevalence of livestock husbandry in LMICs for three major animal taxa: poultry, swine, and ruminants. Microdata relating to ownership of livestock animal species were sourced from various population-based survey programs which together cover the majority of LMICs and categorized. These were georeferenced and spatially matched with a panel of time-fixed environmental and demographic spatial covariates, INLA models were fitted to the resulting database, and probabilities for ownership of each livestock taxon predicted based on the model parameter estimates. The results indicated widespread poultry ownership across rural Central America, the Amazon basin, tropical Africa and river basins and forests of East Asia. Swine husbandry is the least widely practiced among the three livestock taxa and concentrated in an undulating belt of higher prevalence extending from central China, through southeast Asia to Northeastern India, though such predictions in data-sparse regions (particularly Muslim-majority areas) represent regional covariate patterns rather than fine-scale measurements. To address non-stationarity in swine spatial structure, region-specific spatial kernels were implemented. Rearing of ruminant livestock appears widespread across subequatorial Africa, Central Asia, the Gobi Desert, the Himalayas, Mongolia and northern India. The models perform impressively by most standard evaluation metrics, and the patterns in their predictions align with external evidence. The distribution of this important risk factor for infectious disease transmission can be modeled using publicly available data sources to generate plausible and potentially actionable predictions over wide geographic areas and identify regions of high exposure to animal disease reservoirs. The resulting predicted prevalence estimates are made available as supplementary files in GIS-compatible format.
1,5-Anhydrohexitol nucleic acid (HNA) is a promising xeno nucleic acid (XNA) for applications such as aptamers and catalysts, due to its favourable physico-chemical properties. Realizing this potential requires efficient and high-fidelity polymerases capable of processing HNA. A key component are HNA reverse transcriptases that convert HNA into DNA, an essential step in standard SELEX workflows. Although HNA reverse transcriptases have been generated by directed evolution, structural insight is essential to guide further enzyme engineering. Here, we report the 2.8 Å crystal structure of the engineered HNA reverse transcriptase KOD-H4, derived from the B-family DNA polymerase of Thermococcus kodakarensis, captured in a closed ternary complex with dATP, a 3'-terminated primer and a mixed HNA/DNA template. Compared to a previously reported open ternary KOD-H4 structure, the presented structure adopts a more closed conformation with increased finger and thumb domain closure and formation of a canonical Watson-Crick-Franklin base pair at the insertion site. Direct downstream nucleotides show more distorted base pairing and one HNA residue transits from the unusual 1C4 conformation it adopted in the open complex to the 4C1 hexitol sugar conformation. These findings demonstrate that KOD-H4 can form a closed, pre-catalytic complex resembling that of the wildtype enzyme with natural substrates, and reveal state-dependent conformational flexibility of HNA. Such flexibility should be considered in the design and optimization of enzymes that process HNA.