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Single-cell profiling of kinase substrate phosphorylation by single-molecule imaging
Protein phosphorylation regulates diverse cellular processes, yet its analysis at the single-cell level remains challenging due to the low abundance of phosphoproteins. Here, we present a highly sensitive system for profiling phosphorylation of kinase substrates in individual cells. The method integrates fluorescence labeling of single-cell proteomes, immunoprecipitation using antibodies recognizing phosphorylation within specific amino acid motifs, miniaturized SDS-PAGE, and single-molecule detection using a custom-built light-sheet fluorescence microscope. We applied this approach to analyze substrates of casein kinase 2 (CK2) in HeLa cells treated with the phosphatase inhibitor calyculin A. Bulk and pseudo-single-cell analyses confirmed treatment-induced accumulation of phosphorylated CK2 substrates and demonstrated quantitative performance over biologically relevant input ranges. Importantly, true single-cell measurements revealed heterogeneous phosphorylation patterns across molecular weight regions, highlighting cell-to-cell variability in CK2 signaling that is obscured in bulk analyses. This platform enables profiling of the phosphorylation states of a wide range of kinase substrates in individual cells and provides a foundation for dissecting heterogeneous signaling dynamics.
Fasciola hepatica excretory-secretory products attenuate demyelination and reduce neuroinflammation in the Cuprizone –induced multiple sclerosis model
Multiple sclerosis (MS) is characterized by chronic neuroinflammation and progressive demyelination, with current therapies failing to adequately address both processes simultaneously. Helminth-derived excretory–secretory products (ESP) are immunomodulatory molecules that suppress host inflammation and are promising candidates for MS treatment. However, their capacity to promote remyelination remains poorly understood. This study investigated the effects of Fasciola hepatica ESP in the cuprizone-induced demyelination model. Male C57BL/6 mice were divided into four groups: control (CON), control + FES (CON + FES), cuprizone (CUP), and cuprizone + FES (CUP + FES). FES was administered intraperitoneally during the demyelination phase. Motor and cognitive functions were evaluated using open field, rotarod, wire grip, and shuttle box tests. Neuroinflammation was assessed by measuring TNF and IL-1β levels, while myelin-related changes were evaluated by MBP and Olig2 expression (ELISA and qPCR) and Luxol Fast Blue staining. FES treatment significantly reduced pro-inflammatory cytokines, increased MBP and Olig2 levels, improved myelin integrity, and enhanced motor and cognitive performance compared to untreated cuprizone mice, though full recovery to control levels was not achieved. These findings demonstrate that FES attenuates neuroinflammation and is associated with partial improvements in myelin-related outcomes in the cuprizone model, supporting the therapeutic potential of helminth-derived molecules for MS.
Correction: Earliest evidence for invasive mitigation of dental caries by Neanderthals
Gender differences in provider practice characteristics and medicare payment & services among diagnostic radiologists
Background Gender-based differences in representation, practice settings, sub-specialty, billings, and payments among radiologists are poorly understood. Purpose To compare representation, scope of practice, productivity, and Medicare payments between female and male diagnostic radiologists. Methods and materials This cross-sectional (2017–2021) retrospective study examined the CMS Medicare Fee-for-Service Provider Utilization and Payment Database for payments to male and female diagnostic radiologists. The study practice setting, volume of services provided, number of unique beneficiaries served and financial data. Descriptive statistics were reported as mean with standard error (SE) for financial variables and median with interquartile range (IQR) for non-financial variables. Results Between 2017–2021 data, a total of 33,029 diagnostic radiologists provided services to Medicare fee-for-service beneficiaries, of whom 8,217 (24.9%) were female. Female radiologists were disproportionately in academic versus non-academic radiology (34.2% vs 22.6%); practicing in urban versus rural areas (26.3% vs 17.0%); in larger practices (30.1% in practices with ≥100 radiologists vs 17.5% in practices with 1–9 radiologists); and in certain subspecialties (breast imaging- 64.9%). On average, female radiologists received 86% of the total Medicare payments received by male radiologists (mean [SE], $301,931 [$1810] vs $352,604 [$1,164]; p < .001). Female radiologists billed fewer median total services (6,660 vs 9,859; p < .001), served fewer beneficiaries (5,441 vs 7,678; p < .001), and billed fewer unique codes (20 vs 39; p < .001). Overall, female radiologists’ average (SE) payment per service was 18% higher than males [$36.26 (0.13) versus $30.71 (0.06), respectively, but were similar within each subspecialty. Conclusion Female radiologists are disproportionately in academic, urban, and large practices and work as subspecialists. Further, while total Medicare payments received by females were less than males, average payments per service were higher for females overall, but comparable by sex in the same subspecialty.
Size-dependent mechanical performance and defect sensitivity in T4,4,4-graphyne nanosheets: A comprehensive multi-parameter investigation
The mechanical performance of T4,4,4-graphyne was systematically investigated using molecular dynamics (MD) simulations. The study explores the effects of structural size, temperature, defect density, and layer count on key mechanical properties, including elastic modulus, ultimate tensile strength, and toughness. Results reveal a significant size-dependent enhancement in mechanical properties, with increasing nanosheet length leading to higher stiffness, strength, and energy absorption capacity. Conversely, elevated temperatures cause a notable degradation in these properties due to thermal softening and bond weakening. The presence of randomly dispersed vacancy defects severely compromises the material’s mechanical integrity, with even low defect concentrations causing substantial reductions in modulus, strength, and toughness. Additionally, multilayer configurations exhibit improved mechanical behavior compared to monolayers, attributed to enhanced interlayer load transfer and reduced surface effects. The fracture process, analyzed under uniaxial tension, shows brittle failure characteristics with anisotropic crack propagation behavior, consistent with the directional bonding in T4,4,4-graphyne. These findings provide critical insights into the mechanical response of this emerging 2D material, offering valuable guidance for its application in nanoelectromechanical systems, flexible electronics, and high-strength composite materials.
Optimization of milk and coconut milk proportions in the formulation of ice cream and evaluation of its quality characteristics
Perspectives on preconception care in Ethiopia: Social, cultural, and structural determinants
Background Although Ethiopia introduced its first national preconception care (PCC) guideline in 2024, PCC remains rarely integrated into routine practice, and existing studies have largely focused on women’s knowledge and behaviors. Little is known about how adults navigate PCC within broader social, cultural, and structural contexts. This study provides an in-depth urban Ethiopian analysis of how adults experience and negotiate PCC within intersecting gender, moral, and institutional systems, offering insights beyond individual-level understanding. Methods An interpretive description design guided semi-structured interviews with 18 adults (10 women, 8 men; 19–45 years) recruited through maximum-variation sampling from two public hospitals in Addis Ababa. Interviews were conducted in Amharic, transcribed, translated, and analyzed inductively. Data analysis was guided by ID principles, complemented by thematic analysis techniques informed by grounded theory, including line-by-line coding, constant comparison, and analytic memoing. Field notes captured contextual and relational dynamics. Results Seven interrelated themes highlighted complex dynamics in PCC. Knowledge was fragmented and often recognized only after complications, shaped by marital gatekeeping, faith-based beliefs, and exclusion of unmarried women. PCC was valued as protective and morally significant, but stigma, poverty, staff shortages, and inconsistent services constrained practice. Men were largely financial supporters, though many expressed a desire to participate, limited by gender norms and women-centered services. Pharmacies and digital media provide informal but sometimes unsafe guidance. Emotional experiences, fear, guilt, secrecy, and hope were central to PCC engagement. Education, peer influence, schools, and community leaders emerged as catalysts for uptake, yet participants emphasized that sustainable PCC required visible institutional support, reliable services, and government recognition. Strategies to enhance practice included simplifying communication, creating accessible clinic entry points, and mobilizing community networks to normalize pre-pregnancy preparation. Conclusions This study reveals PCC in urban Ethiopia as a socially negotiated, morally contested, and structurally uneven practice, far more complex than knowledge deficits imply. These findings offer novel, actionable direction for implementing Ethiopia’s PCC guideline through visible, inclusive, relational, and community-anchored approaches that address the social conditions shaping PCC access.
Exposure to violence in public places among youths with a refugee background and their Swedish-born peers
Abstract Youths constitute a large proportion of forcibly displaced individuals worldwide. Although exposure to violence in countries of origin and during migration is well documented, less is known about its persistence in resettlement contexts, particularly regarding violence in public places (ViPP). This study aimed to examine youths’ perceived societal safety, exposure to ViPP in Sweden, and differences between participants with and without a refugee background. The study included 79 youths with a refugee background (M = 21.4 years, SD = 2.3, 54.4% male) and 105 Swedish-born peers (M = 21.5 years, SD = 2.4, 50.9% male). Data were collected between 2019 and 2022 using a questionnaire developed through a participatory approach. Fisher’s exact test was used for quantitative analyses, and content analysis was applied to qualitative data. A total of 33.3% reported experiences of ViPP in Sweden. Despite descriptive differences between groups, no statistically significant differences were observed in direct exposure to ViPP between participants with and without a refugee background. However, participants with a refugee background reported significantly higher levels of perceived unsafety and greater restrictions in their daily lives. Qualitative responses indicated that increased education, surveillance, and adult presence could improve public safety. These findings highlight the need to ensure the safety of youths in public places, particularly by addressing perceived unsafety among young people with a refugee background.
Negotiating illness through faith: Religious narratives of cancer etiology among patients and caregivers in Addis Ababa, Ethiopia
Cancer has become a major global public health threat, and individuals diagnosed with cancer and their family caregivers often seek to understand their illness experiences. Religious, spiritual, and sociocultural beliefs play a central role in shaping illness interpretations and care-seeking behaviors. However, in Ethiopia, limited attention has been paid to how these frameworks influence cancer experiences. This study explored religious narratives and interpretations of illness etiology among individuals diagnosed with cancer and their family caregivers and examined their influence on illness experiences and health-seeking practices. An interpretive phenomenological approach was employed using semi-structured, in-depth interviews with 41 participants, including individuals with cancer, family caregivers, and religious leaders. Data were transcribed, translated, and thematically analyzed to examine the processes of meaning-making. The findings showed that participants commonly conceptualized addiction as spiritually mediated rather than merely biologically determined. Illness was frequently interpreted as having a divine or supernatural origin, including punishment, a test of faith, God’s will, or spiritual affliction. These interpretations were dynamic and shaped by religious teachings, sociocultural contexts and personal reflections. Religious frameworks influence emotional responses, coping strategies, and health-seeking behaviors, providing comfort and resilience while shaping treatment decisions. They functioned not only as explanatory frameworks for illness but also as practical resources that structured participants’ responses to and negotiations with uncertainty, suffering, and responsibility. Religious meaning-making plays a central role in illness interpretations and overall experiences in cancer trajectories. Recognizing and engaging with these spiritual frameworks may enhance culturally responsive and patient-centered cancer care. Integrating spiritual sensitivity into clinical practice and collaborating with religious leaders may improve communication, trust, and psychosocial support for patients and their families.
Clinical outcomes and healthcare utilization in very elderly patients with pneumonia: an analysis of korean national health insurance claims database
Correlation of single-round strokes & performance indicators: Paris Olympic elite female golfers
Despite golf’s Olympic reinstatement, quantitative research on elite female Olympic golfers remains scarce, with most studies focusing on tour or amateur players. This study aimed to quantify relationships between single-round strokes and 13 performance indicators among top 10 2024 Paris Olympic female golfers (including ties), identify key predictors, and build a predictive model using correlation analysis, principal component analysis (PCA) and LASSO regression with leave-one-out cross-validation (LOOCV) and bootstrap validation to address overfitting concerns inherent to small-sample analyses. Results showed single-round strokes correlated negatively with GIR ( r = −0.484), SG: Putting ( r = −0.567) (both p < 0.01) and positively with Putt-Total ( r = 0.582, p < 0.01). PCA extracted five principal components explaining 77.94% of total variance, representing distinct performance constructs. LASSO regression identified seven stable predictors (bootstrap selection frequency ≥ 90%): Greens in Regulation (100%), Putt-Total (99%), SG: Approach the Green (98%), SG: Around the Green (97%), SG: Putting (94%), SG: Off the Tee (92%), and Scrambling (88%). Model performance yielded an adjusted R² of 0.928 with cross-validated MSE of 0.528 based on 48 rounds from 12 golfers, though external validation is warranted. This study fills Olympic golf research gaps, providing evidence for targeted training and tournament strategy optimization.
Comparison of personal portable and fixed-site PM2.5 monitoring strategies in predicting pulmonary function decline among asthmatic children in Seoul, Korea: a pilot study
Targeted Muscle Reinnervation (TMR) or Regenerative Peripheral Nerve Interface (RPNI) for pain prevention in patients with limb amputation: A protocol for a systematic review and meta-analysis
Introduction Regenerative Peripheral Nerve Interface (RPNI) and Targeted Muscle Reinnervation (TMR) have demonstrated superior outcomes compared to classical amputation in prophylactic prevention of pain, primarily by reducing the incidence of symptomatic neuromas, residual limb pain, and phantom limb pain. However, direct comparisons between these two techniques remain limited. Furthermore, their comparative effectiveness across diverse patient demographics (including age, sex, and comorbidities) and surgical variables (amputation level, etiology, and nerve handling) has not been systematically evaluated. Therefore, the objective of this systematic review and meta-analysis is to synthesize the available evidence to determine the comparative safety and efficacy of primary TMR or RPNI. Methods and design This review will be conducted following the methodological guidance of the Cochrane Handbook for Systematic Reviews of Interventions . A comprehensive electronic search will be performed in the Cochrane Central Register of Controlled Trials (CENTRAL), Web of Science, Scopus, PubMed, and MedRxiv, without language restrictions. We will include randomized controlled trials, quasi-randomized trials, and observational studies. Study selection and data extraction will be managed using Covidence. Two reviewers will independently screen titles and abstracts, assess full-text eligibility, evaluate risk of bias, and extract data. Ethics and dissemination As this systematic review relies on the analysis of secondary data from published studies, ethical approval is not required. Findings will be disseminated through publication in a peer-reviewed journal and presented at relevant conferences. PROSPERO registration number CRD42024617299
Intelligent emotion and behavior recognition for online learners using deep affective computing
Vertical distribution and migration of microplastics in soils from Fars Province, Southwest Iran
Microplastics (MPs) in soils are becoming increasingly recognised as important terrestrial contaminants, yet their vertical distribution below the plough layer remains poorly understood. Here, we examined 76 genetic soil horizons from 27 profiles along four transects spanning managed and unmanaged land uses in Fars Province, southwest Iran, to depths of up to 140 cm. A total of 392 MPs were recovered from 71 of 76 samples, comprising 342 fibres (87.2%), 48 fragments or films and two spherules, with concentrations reaching about 320 particles kg -1 of dry soil. Raman analysis of 98 representative particles showed a dominance of polyamides, polyesters and polyolefins. MP abundance, size and polymer type were heterogeneous among sites and horizons, and only a few statistically significant relationships with soil properties were observed (notably inverse relationships with sand at Darab and silt at Sarvestan). MPs occurred throughout the profiles, including the deepest horizons, with no consistent size- or polymer-related fractionation with depth. These observations indicate sustained MP accumulation in soils and suggest that downward transport can occur under arid to semi-humid conditions through percolation, drying-cracking of the substrate and bioturbation. Because strong local point sources were not evident at most sites and fibres dominated the assemblage, atmospheric deposition is considered a major contributor at the soil surface, although additional source-specific data are needed to confirm this inference. The persistence and vertical mobility of MPs in Fars soils indicate potential risks to subterranean ecosystems and groundwater quality.
Tumor probability mapping of fluorescence intensity during image-guided surgery of head and neck cancer
Abstract Achieving negative surgical margins remains a critical determinant of local recurrence and survival in head and neck cancer (HNC) surgery. Current intraoperative margin assessment techniques, including frozen section analysis, suffer from sampling errors and procedural delays. Tumor-targeted fluorescence imaging offers real-time tumor visualization but lacks standardized quantitative approaches for clinical decision-making. We developed a Tumor Probability Mapping (TPM) framework using panitumumab-IRDye800 fluorescence imaging in 16 HNC patients. Ex vivo specimens and gross tissue sections were imaged using near-infrared fluorescence systems. A total of 5,442 regions of interest (ROIs) were manually distributed across fluorescence images of gross specimen sections validated by histopathology. Signal-to-background ratios (SBR) were calculated and used to train the following predictive models: generalized linear model fit standard logistic regression (MATLAB, glmfit), standard logistic regression (R, LOG), mixed-effects logistic regression (GLMER), and Bayesian mixed-effects regression (BRMS). Model performance was evaluated using receiver operating characteristic and area under the curve (ROC-AUC) analysis, sensitivity, specificity, along with beta-calibration and model fit. All models demonstrated excellent (> 90%) discriminative ability between tumor and normal tissue. The glmfit model, selected for clinical implementation, achieved 95.8% accuracy, 90.8% sensitivity, 98.8% specificity, and an AUC of 0.989 on test data. The final TPM algorithm provides real-time probability assessment of tumor presence based on fluorescence intensity quantified by histopathology validated historical data. TPM represents a significant advancement in fluorescence-guided surgery by converting qualitative fluorescence signals into quantitative probability assessments validated against histopathology. This approach provides surgeons with standardized, real-time tumor probability information that extends beyond qualitative assessments and/or binary threshold determinations, potentially improving surgical outcomes by enhancing margin assessment and reducing local recurrence rates.
Cumulative metabolic stress (microfilarial infection + moult) constrains the expression of carotenoid-based honest signals in breeding male village weavers (Ploceus cucullatus) of Amurum Forest Reserve, Nigeria
In wild birds, the breeding season involves a convergence of metabolically demanding life-history stages, including reproduction, moult, and immune defense. We investigated the relationships between microfilarial infection, moult, redox homeostasis, and plumage quality in breeding male village weavers (Ploceus cucullatus) at the Amurum Forest Reserve, Nigeria. We compared four groups (n = 148 total) sampled within 3 mins post-capture: infected-moulting (IM), infected-non-moulting (IN), non-infected-moulting (NM), and non-infected-non-moulting (NN). Physiological condition was assessed using the erythrocyte glutathione ratio (GSH:GSSG) and circulating glucose, while plumage reflectance traits were integrated into a composite quality axis (PC1). Microfilarial infections were present in 52.0% (n = 77) of individuals; mean parasite intensities were 6.13 ± 0.35 mf/µL (IN) and 6.45 ± 0.41 mf/µL (IM). Physiological indices varied strongly across groups. The GSH:GSSG ratio was reduced in infected birds, indicating altered redox balance (rs = −0.65). Circulating glucose was highest in the infected non-moulting group (IN) but substantially reduced in the infected moulting group (IM). Across physiological and ornamental traits, individuals experiencing both infection and moult (IM group) exhibited the strongest reductions relative to all other groups. However, this pattern reflects a statistically supported Infection × Moult interaction, rather than an untested synergistic or non-linear effect, as evidenced by significant IN vs. IM contrasts in glucose (Table 3; z = 33.43, P < 0.0001, d = 6.10) and plumage quality. This interaction was associated with reduced plumage hue and lower integrated signal quality. Our results suggest that microfilarial infection and moult impose overlapping physiological demands that constrain metabolic regulation and the expression of carotenoid-based ornaments. Intracellular redox balance emerges as a potential mechanistic link between physiological state and ornamental expression, supporting the hypothesis that village weaver plumage reflects variation in sustained physiological condition under natural ecological stress.
A novel-based Kolmogorov-Arnold Networks for high accuracy mapping of water and vegetation: a study of the global main rivers
Neutrophil extracellular traps in the animal model of adenine-induced chronic kidney disease
Neutrophil extracellular traps (NETs) are suggested to play a role in chronic kidney disease (CKD). Whether they are indeed involved in the pathogenesis of animal models of CKD has not been proved. This study tested the hypothesis that a genetic deficiency of peptidylarginine deiminase 4 (PAD4) – a regulator of NETs production will protect mice from developing adenine-induced nephropathy as a model of CKD. Adult male Padi4 −/− mice and their wild-type counterparts received 100 mg/kg adenine or saline i.p. daily for 14 days. Markers of renal function and NETs-related biomarkers were assessed during the CKD induction in plasma and urine. In wild-type mice, adenine injections decreased body weight by 20% and increased plasma creatinine (twofold), neutrophil gelatinase-associated lipocalin (21-fold), and neutrophil elastase (4-fold). No significant differences were found between the Padi4 −/− and wild-type mice except the earlier increase in plasma creatinine in Padi4 −/− mice (day 3 vs. day 7). Analyses of neutrophil elastase and myeloperoxidase in plasma and urine suggest that neutrophils are activated in adenine-induced nephropathy, but the production of NETs seems not to be directly involved in the pathogenesis of this CKD model. Further studies should clear the role of NETs in other kidney disease models with clinical relevance.