Endometrial health is a key determinant of female fertility and successful pregnancy outcomes, making the accurate diagnosis of endometrial lesions essential for the success of assisted reproductive technology. While hysteroscopy remains the gold standard for uterine cavity evaluation, interpretation can vary based on clinical expertise. To address this, we developed a deep learning-based clinical decision support system to classify hysteroscopic images from high-resolution (4 K) videos into three categories: normal endometrium, endometrial polyps, and endometritis. Endometritis cases were classified based on hysteroscopic features suggestive of inflammation; no histopathological confirmation was obtained. Utilizing a dataset of 1500 expert-annotated images from 200 clinical videos, we applied transfer learning across four architectures: VGG-16, VGG-19, DenseNet-121, and EfficientNet-B0. Our results show that the models achieved classification accuracies between 85 and 89%, with DenseNet-121 demonstrating superior performance, specifically achieving a sensitivity of 93% and an AUC of 98.8% for polyp detection, alongside a precision of 90% for endometritis. Furthermore, Grad-CAM visualization confirmed that the networks focused on clinically relevant morphological features, enhancing model interpretability. These findings suggest that deep learning may serve as a supportive tool to assist clinicians in hysteroscopic analysis, pending validation on external datasets and with pathologically confirmed labels.
We report observations of an energetic-particle mode (EPM) in a linear plasma device that corresponds to an axial bounce resonance of fast ions sourced from neutral-beam injection (NBI). In our experiment, novel dynamic shaping of a magnetic mirror field reduces the distance between the turning points of fast ions while introducing a minimum in the expected bounce frequency; this method decouples the plasma length from the fast-ion motion. We observe an axisymmetric magnetic mode with frequency evolution and axial extent that matches the predicted bounce motion. This experiment is repeated for two different NBI configurations with distinct bounce motions, thus confirming the mode's origin as an axial bounce resonance. We then show that the EPM persists during a transition from a magnetic mirror to a field-reversed configuration (FRC), making this the first observation of an axial bounce resonance in an FRC. Additionally, we use the spatial structure of the EPM to diagnose the change in magnetic topology from mirror to FRC. Finally, a 2D simulation of the FRC formation accurately models the experimental observations and confirms that the mode arises from the clustering of resonant fast ions. These results have insightful implications for the control and design of linear fusion devices with substantial populations of fast ions.
Severe pertussis in children can deteriorate rapidly, and early identification of patients likely to require intensive care remains challenging. We aimed to develop and internally validate an admission-time prediction model for ICU admission among hospitalized children with PCR-confirmed pertussis. We conducted a single-center retrospective prediction-model study based on a hospitalized cohort at a provincial pediatric infectious disease center. Consecutive hospitalized children with PCR-confirmed pertussis between 6 June 2020 and 24 June 2025 were screened. Two children were excluded because admission white blood cell count (WBC), a prespecified core predictor, was unavailable in the retrievable admission-time record, leaving 662 children aged 24 days to 14 years for analysis. The primary outcome was ICU admission, defined a priori as ICU length of stay > 0 days. We prespecified a multivariable logistic regression model using seven admission-time predictors: age (months), sex, symptom duration before admission (days), paroxysmal cyanosis documented at presentation, imaging consolidation/atelectasis, admission WBC scaled per 5 × 109 /L, and any recorded pertussis-containing vaccine dose. Internal validation used 5-fold cross-validation with out-of-fold predictions; model performance was assessed by discrimination (AUC), overall prediction error (Brier score), calibration, and decision curve analysis. ICU admission occurred in 61/662 (9.2%) children. Compared with non-ICU patients, ICU-admitted children were younger (median 2.0 vs 60.0 months) and had higher admission WBC (median 18.7 vs 10.0 × 109 /L). Paroxysmal cyanosis (49.2% vs 6.0%) and imaging consolidation/atelectasis (23.0% vs 10.5%) were more frequent, whereas recorded vaccination was less common (23.0% vs 87.9%). On internal validation, the model showed excellent discrimination (AUC 0.953; 95% CI 0.933-0.972) with low overall prediction error (Brier score 0.045). Calibration was good (intercept -0.017; slope 0.992), and decision curve analysis suggested positive net benefit over clinically relevant threshold probabilities. An admission-time multivariable model showed good internal performance for predicting ICU admission in pediatric pertussis and may support early risk stratification. External validation is required before routine implementation.
Delirium is a frequent manifestation of acute brain dysfunction in critically ill patients with bloodstream infections (BSI). While the association between sepsis and delirium is well-established, pathogen-specific neurotoxic effects, particularly those induced by Staphylococcus aureus (S. aureus), remain inadequately elucidated. This study aimed to evaluate the independent association between S. aureus BSI and the risk of 7-day incident delirium in critically ill adults. A retrospective cohort study was conducted using the MIMIC-IV database, including 3,226 adult patients with confirmed BSI. To minimize confounding bias, a "doubly robust" estimation approach was employed, combining Inverse Probability of Treatment Weighting based on Covariate Balancing Propensity Scores (IPTW-CBPS) with multivariable adjustment. Fine-Gray proportional subdistribution hazard models were utilized to assess the association between pathogen status and 7-day delirium, accounting for death as a competing risk. Explainable machine learning using the eXtreme Gradient Boosting (XGBoost) algorithm and Shapley Additive exPlanations (SHAP) value analysis was performed to identify feature importance. Sensitivity analyses for the 3-day acute risk and methicillin-resistant Staphylococcus aureus (MRSA) phenotype were also conducted. Among 3,226 patients, 618 (19.2%) were identified with S. aureus BSI. In the fully adjusted Fine-Gray model, compared with Gram-negative infections, S. aureus BSI was robustly and independently associated with an increased risk of 7-day incident delirium (sHR 1.39; 95% CI 1.11-1.74; p = 0.005). This association was even more pronounced within the 3-day acute window (sHR 1.60; 95% CI 1.27-2.02; p < 0.001). Conversely, compared with Gram-negative BSI, the MRSA phenotype showed no significant independent association with delirium risk (sHR 1.12; 95% CI 0.81-1.55; p = 0.488). SHAP analysis identified S. aureus infection as the fourth most influential feature for delirium, surpassing age and sedative exposure. Subgroup analyses revealed that the pathogen-specific impact was significantly more prominent in patients with lower baseline illness severity (SOFA < 5) and those without a history of dementia (P for interaction < 0.001 and 0.004, respectively). SHAP value analysis further confirmed this finding by demonstrating a higher risk contribution of S. aureus in patients with lower SOFA scores. S. aureus BSI is a robust independent risk factor for 7-day incident delirium compared with Gram-negative BSI, with the hazard being more pronounced within the first 3 days of infection. Notably, compared with Gram-negative BSI, the MRSA phenotype was not independently associated with delirium risk. Identifying S. aureus as a critical driver of early brain dysfunction provides a pathogen-specific framework for risk stratification. Consequently, clinicians should prioritize early neurocognitive monitoring and targeted neuroprotection (e.g. the ABCDEF bundle) for all patients with S. aureus infection.
Menke-Hennekam syndrome (MKHK) is a rare autosomal dominant disorder caused by mutations in the CREBBP and EP300 genes. The absence of established diagnostic criteria and non-specific clinical manifestations complicate timely diagnosis and management. This report presents a case of MKHK in which early diagnosis and intervention were achieved through the application of rapid whole-genome sequencing (rWGS), a tool that offers superior speed and genomic coverage compared to whole-exome sequencing (WES). This case report describes a male Han Chinese neonate who presented at birth (0 days) with intrauterine growth restriction, respiratory distress, and feeding difficulties. During follow-up, he developed hearing loss and demonstrated global developmental delay. Clinical examination revealed craniofacial dysmorphism. Trio rWGS was performed in the neonatal period, with results returned within 72 h of sample submission at 23 days of age. Trio rWGS identified a de novo missense variant in the CREBBP gene (c.5570A > C, p.His1857Pro). Sanger sequencing confirmed its absence in both parents, and the variant was classified as likely pathogenic despite no prior documented cases. Based on integrated genetic and clinical findings, a neonatal diagnosis of MKHK-ID4 was established. Following this diagnosis, early targeted interventions were initiated, including hearing aid fitting, enrollment in a comprehensive rehabilitation program, and planning for necessary surgical corrections. Significant developmental improvement was observed at the 15-month follow-up assessment. In this case, rWGS facilitated a neonatal diagnosis of MKHK-ID4 and enabled early multidisciplinary intervention during a critical neurodevelopmental window. This experience suggests that such an approach may contribute to improved developmental outcomes in this rare disorder, though further studies are required to confirm its broader applicability and long-term benefits.
ObjectiveThis study aimed to systematically investigate the independent predictive value of the platelet-to-neutrophil ratio and platelet-to-lymphocyte ratio for 28-day all-cause mortality in patients with sepsis and to further evaluate their incremental discriminatory capacity when added to conventional prognostic scoring models.MethodsThis single-center retrospective cohort study involved 287 adult patients diagnosed with sepsis according to the Sepsis-3 criteria. Patients were stratified into high- and low-level groups based on the median values of platelet-to-neutrophil ratio and platelet-to-lymphocyte ratio, and baseline characteristics, and clinical outcomes were compared between groups. Univariate and multivariate Cox proportional hazards regression models were used to assess the independent associations of platelet-to-neutrophil ratio and platelet-to-lymphocyte ratio with 28-day mortality risk. To evaluate predictive performance, multiple models were constructed and compared, including Sequential Organ Failure Assessment score alone, Acute Physiology and Chronic Health Evaluation II score alone, each combined separately with platelet-to-neutrophil ratio or platelet-to-lymphocyte ratio, and a composite model integrating Acute Physiology and Chronic Health Evaluation II, day-3 lactate level, platelet-to-neutrophil ratio, and platelet-to-lymphocyte ratio. Model performance was rigorously assessed using receiver operating characteristic curves, calibration plots, decision curve analysis, integrated discrimination improvement, and net reclassification improvement, enabling a comprehensive evaluation of discrimination, calibration, clinical utility, and the incremental value of novel biomarkers.ResultsMultivariate Cox regression analysis revealed that lower platelet-to-neutrophil ratio (hazard ratio = 0.97, 95% confidence interval: 0.95-0.99, p = 0.012) and higher platelet-to-lymphocyte ratio (hazard ratio = 1.01, 95% confidence interval: 1.01-1.01, p = 0.043) were independently associated with increased 28-day all-cause mortality. Kaplan-Meier survival analysis confirmed significantly higher cumulative mortality in the low-platelet-to-neutrophil ratio and high-platelet-to-lymphocyte ratio groups (log-rank p < 0.05). The composite model incorporating Acute Physiology and Chronic Health Evaluation II, day-3 lactate level, platelet-to-neutrophil ratio, and platelet-to-lymphocyte ratio demonstrated superior predictive accuracy, with an area under the receiver operating characteristic curve (area under the curve) of 0.824 (95% confidence interval: 0.778-0.871), significantly outperforming both the Acute Physiology and Chronic Health Evaluation II only model (area under the curve: 0.746, p < 0.001) and the Sequential Organ Failure Assessment only model (area under the curve: 0.667, p < 0.001). Decision curve analysis showed that this model provided greater clinical net benefit across a broad range of threshold probabilities. Furthermore, both integrated discrimination improvement and net reclassification improvement analyses confirmed statistically significant improvements in discrimination and reclassification accuracy after the inclusion of platelet-to-neutrophil ratio and platelet-to-lymphocyte ratio (both p < 0.001).ConclusionPlatelet-to-neutrophil ratio and platelet-to-lymphocyte ratio are independent predictors of 28-day mortality in patients with sepsis. Incorporating these readily available and cost-effective inflammatory markers into traditional prognostic systems-such as Acute Physiology and Chronic Health Evaluation II-and combining them with dynamic lactate monitoring substantially enhances the discrimination, calibration, and clinical utility of risk prediction models. These findings provide robust evidence supporting the use of platelet-to-neutrophil ratio and platelet-to-lymphocyte ratio for early risk stratification and individualized prognostic assessment in clinical practice.
To prepare hydroxyapatite/chitosan (HA/CTS) nanocomposites at four weight ratios (85/15, 70/30, 50/50, 30/70) and two concentrations (3 and 5 wt.%), incorporate them into conventional glass ionomer cement (GIC), and evaluate their effect on compressive strength, shear bond strength to dentin, and antibacterial activity. HA/CTS nanocomposites were synthesized by co-precipitation and characterized by TEM and FTIR. Nine groups were prepared (n = 10): one unmodified control (Group I) and eight modified groups (Groups II-IX). Compressive strength was tested per ISO 9917-1:2007; shear bond strength to human dentin and antibacterial activity (agar diffusion against Streptococcus mutans, Staphylococcus aureus, and Escherichia coli) were also evaluated. One-way and two-way ANOVA with Tukey's HSD were applied (P ≤ 0.05). TEM confirmed nanoscale particles (7-50 nm) with dispersion improving at higher chitosan content. FTIR verified dual-phase incorporation and Ca2+-NH2 coordination bonding. Compressive strength (F(8,81) = 177.509; P < 0.0001) and shear bond strength (F(8,81) = 202.262; P < 0.0001) differed significantly among groups. A significant ratio × concentration interaction was confirmed for both properties (P < 0.0001). Only the 70/30 nanocomposite at 3 wt.% exceeded the control compressive strength (133.44 vs. 115.02 MPa; P < 0.05). All eight modified groups showed significantly higher shear bond strength than the control (6.04 MPa), with the 85/15 at 5 wt.% achieving the highest value (13.30 MPa). No inhibition zones were detected in any group. The 70/30 HA/CTS nanocomposite at 3 wt.% optimizes compressive strength, while the 85/15 at 5 wt.% optimizes dentin adhesion. Overall, the 70/30 HA/CTS nanocomposite at 3 wt.% demonstrated the most favorable balanced performance profile across both mechanical outcomes. The absence of detectable antibacterial activity under the present agar diffusion testing conditions may be related to chitosan immobilization within the nanocomposite matrix and the diffusion limitations of the assay method. Findings suggest the potential for application-specific optimization of HA/CTS-modified GIC under controlled in vitro conditions, pending further long-term and in vivo validation. HA/CTS nanocomposite modification enhanced the mechanical and adhesive performance of conventional GIC under the present experimental conditions. The observed improvement in compressive and shear bond strength across the modified formulations may support improved marginal integrity in esthetic GIC restorations. Two application-specific optimal formulations were identified based on the functional demands of the restoration site, with the 70/30 HA/CTS nanocomposite at 3 wt.% representing the most favorable overall formulation combining enhanced compressive strength with clinically acceptable dentin adhesion; however, further long-term and in vivo validation remains necessary before definitive clinical translation.
Reliable and valid assessment tools are critical for evaluating and improving primary school students' hand hygiene behaviors, yet corresponding adaptable assessment methods are currently limited. We aimed to develop and validate a Hand Hygiene Behavior Questionnaire for Primary School Students (HHBQ-PSS). Cluster sampling recruited 376 Grade 4-5 students from two central primary schools. The 15-item HHBQ-PSS encompasses three dimensions (capability, opportunity, motivation) of hand hygiene, using a 5-point Likert scale. HHBQ-PSS was evaluated through internal consistency reliability, split-half reliability, exploratory factor analysis (EFA), and confirmatory factor analysis (CFA). The Cronbach's α coefficient of HHBQ-PSS was 0.844, and all Cronbach's α coefficients remained above 0.750 after corresponding dimensions were deleted. The Spearman-Brown coefficient was 0.784. EFA extracted four common factors (cumulative variance contribution rate = 58.67%). CFA confirmed adequate model fit (χ2/df = 2.832, CFI = 0.905, GFI = 0.927, AGFI = 0.893, and RMSEA = 0.070). The reliable and valid HHBQ-PSS may inform the implementation of targeted health interventions and the improvement of hand hygiene levels among primary school students. HHBQ-PSS demonstrates good reliability and validity to assess hand hygiene behaviors in Chinese primary school students.
Obesity is associated with metabolic dysfunction-associated steatotic liver disease (MASLD), for which drugs and nutraceuticals are being developed. We investigated whether camelina seeds from the Brassicaceae family and their components can attenuate metabolic disorders in animal models of genetic obesity, including fatty liver. In two separate 5-week experiments, female obese (fa/fa) Zucker rats and female db/db mice were used. By appropriately incorporating camelina seeds or their oil into the diet, we investigated, in the rat model, the effects of the oil and non-oil fractions on intestinal and liver function, as well as on lipid metabolism. Subsequently, in a mouse model, oral administration of glucocamelinin, the main camelina glucosinolate, was performed to determine whether this group of compounds might be responsible for the observed beneficial effects. Dietary camelina seeds counteracted liver hypertrophy and steatosis in obese rats, as confirmed by macroscopic and histological images, along with a several-fold lower content of total fat, triglycerides, and cholesterol (p < 0.01 for each). The rat experiment indicated that the non-oil fraction of camelina seeds was responsible for these beneficial effects. Oral glucocamelinin attenuated liver hypertrophy and steatosis in obese mice, as confirmed by macroscopic and histological images, along with considerably lower total fat (p < 0.05) and cholesterol (p < 0.01) contents. In both obese rats and mice, these and other beneficial effects were associated with alterations in the hepatic expression of genes crucial to lipid and glucose metabolism, including a higher expression of those encoding thyroid hormone receptor β (THR-β; p < 0.01 and p < 0.05) and fibroblast growth factor 21 (FGF21; p < 0.01 and p < 0.05). These findings indicate that camelina seeds may be considered both a dietary component and a source of bioactive glucosinolates relevant to MASLD. The mechanism behind the lipid-lowering effect of glucocamelinin may involve hepatic activation of FGF21 signaling through THR-β.
Managing small Lung-RADS 4 pulmonary nodules is clinically challenging. While CT-guided TTFNAB is standard, its efficacy and safety for lesions ≤2 cm require further evaluation to optimize outcomes and reduce unnecessary surgeries. This study aimed to evaluate the diagnostic accuracy, safety, and factors influencing the outcomes of CT-guided TTFNAB in Lung-RADS category 4 pulmonary nodules measuring ≤2 cm. Retrospective study. Single-center, a tertiary referral center. A retrospective analysis was performed on 95 patients who underwent CT-guided TTFNAB between January 2021 and April 2024. Data included demographics, lesion characteristics (size, location, density, and pleural proximity), histopathological findings, and procedural outcomes. The primary endpoints were diagnostic accuracy, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and factors influencing TTFNAB results, alongside procedural complications-specifically pneumothorax and chest tube requirement. 95 patients. Of the 95 nodules, 26 (27%) were benign and 69 (73%) were malignant according to TTFNAB. Final pathology confirmed 18 (19%) benign and 77 (81%) malignant lesions. All biopsies yielded sufficient material. Sensitivity, specificity, and overall diagnostic accuracy were 83%, 94%, and 89%, respectively. The PPV was 98%, while the NPV was 65%. Pleural thickening was significantly more frequent in both the TTFNAB benign group (38% vs. 13%, P=.006) and final benign diagnosis group (44% vs. 14%, P=.008). Pneumothorax occurred in 30% of cases, with 24% of these requiring chest tube drainage. No significant associations were found between diagnostic accuracy or complications and variables such as age, emphysema, Lung-RADS category, or nodule size. CT-guided TTFNAB is an effective diagnostic method for confirming malignancy in pulmonary nodules ≤2 cm classified as Lung-RADS 4, providing an overall accuracy of 89% and a PPV of 98%. However, a benign TTFNAB result cannot be considered diagnostically safe: the NPV of 65% and false-negative rate of 35% indicate that a negative biopsy does not reliably exclude malignancy. Clinicians must not rely on a negative result alone; close radiological follow-up or surgical biopsy is essential for these high-risk lesions. The retrospective single-center design, small sample size, and limited generalizability. 2024/010.99/6/23 (Date: 26.07.2024).
Evidence on COVID-19 among people living with HIV (PLWH) remains mixed and context-dependent. We prospectively followed 64 PLWH receiving antiretroviral therapy at a tertiary university hospital in Martin, Slovakia (September 2020-September 2021). SARS-CoV-2 infection was confirmed by RT-PCR. COVID-19 was diagnosed in 54 of 64 participants (84.4%) and was generally mild; none required intensive care, and no deaths were reported. There were no statistically significant differences between PLWH who did and did not develop COVID-19 with respect to age, duration of HIV infection, CD4 cell count, or viral suppression. Participants who developed COVID-19 were more likely to have at least one documented comorbidity; however, this did not appear to translate into severe outcomes in this small cohort. Among those with COVID-19, exploratory comparisons showed no clear differences in short-term outcomes according to mRNA vaccination status. In this small exploratory cohort of ART-treated individuals in Slovakia, we did not identify clear clinical or HIV-related predictors of COVID-19 outcomes; however, the study was underpowered and the findings should be interpreted cautiously.
This study aimed to investigate the clinical utility of the C-reactive protein/albumin ratio (CAR) and the neutrophil/lymphocyte ratio (NLR) in evaluating the severity of sepsis in pediatric patients. A retrospective cohort study was conducted involving 174 pediatric patients hospitalized between June 2022 and November 2025. Sepsis and septic shock were defined according to the 2024 Phoenix criteria. Patients were stratified into a sepsis group (n = 123) and a septic shock group (n = 51). CAR and NLR were calculated from blood samples collected within 24 h of admission. A two-sided p < 0.05 was considered statistically significant. Independent associated factors for septic shock were identified using binary logistic regression analysis. Receiver operating characteristic (ROC) curves were constructed, and the area under the curve (AUC) was calculated to evaluate the diagnostic performance of CAR, NLR, and other relevant biomarkers.The study was conducted in accordance with the STROBE guidelines. Univariate analysis revealed significantly elevated white blood cell (WBC) count, CAR, and NLR levels in the septic shock group compared to the sepsis group (all p < 0.05). Binary logistic regression analysis confirmed that elevated CAR (OR = 2.893, p < 0.001) and NLR (OR = 1.232, p < 0.001) were independent associated factors for progression to septic shock. ROC curve analysis demonstrated that a model combining CAR and NLR exhibited superior discriminative performance compared to either biomarker alone, with a higher AUC (0.802), sensitivity (74.50%), and specificity (74.80%). CAR and NLR appear to be significant independent biomarkers for assessing severity and predicting septic shock in children with sepsis. The combination of CAR and NLR provides improved predictive accuracy compared with either marker alone, suggesting its potential utility in guiding early clinical decisions.
To study the effect of Body mass index (BMI) on 24-h urine quantitative analysis in pediatric urolithiasis, and to explore whether obesity and overweight promote the formation of urinary calculi in children. A comprehensive search of EMBASE, Pubmed and the Cochrane Library, Web of Science, and Scopus were conducted in March 2024 and updated in October 2025 to find all related studies. BMI was used to specify the body size. And then, a predetermined inclusion and exclusion criteria was used to screen each article. Data from appropriate studies was extracted, a meta-analysis was performed using Stata14.0 software. Eight studies, including 1033 children with urolithiasis who underwent 24-hour urine collection for chemical analysis, were included in this meta-analysis. The BMI ≥85th percentile group exhibited significantly higher uric acid excretion (SMD = 0.756, 95% CI = 0.092-1.420, P = .026). No statistically significant differences were found in calcium (SMD = -0.320, 95% CI = -0.600 to -0.050, P = .519), 24-h urine volume (SMD = -0.310, 95% CI = -0.790 to 0.160, P = .555), magnesium (SMD = -0.470, 95% CI = -0.820 to -0.120, P = .471), phosphate (SMD = -0.360, 95% CI = -0.610 to -0.110, P = .805), oxalate (SMD = -0.110, 95% CI = -0.390 to 0.170, P = .315), citrate (SMD = -0.190, 95% CI = -0.680 to 0.290, P = .057), or sodium (SMD = 0.200, 95% CI = -0.390 to 0.800, P = .050) excretion between the two groups. Sensitivity analyses confirmed the robustness of these findings. While overweight/obese children with urolithiasis demonstrate higher urinary uric acid excretion, the absence of significant differences in other key urinary risk factors suggests that BMI alone should not be considered a separate and definitive risk factor for pediatric urolithiasis.
Monosialotetrahexosylganglioside (GM1), a major ganglioside component, is expressed in the spinal cords of mammals. Gangliosides have been shown to play important roles in various physiological processes, including neuronal protection, neurorepair, and stem cell differentiation. Bone marrow-derived mesenchymal stem cells (bMSCs) were expected to be a good source of cells for clinical transplantation and repair due to their abundance, easy accessibility and lack of ethical restrictions. They can also be induced to differentiate into neurons. However, there is still no ideal delivery material or method that allows the sustained and controlled release of GM1 and bMSCs for clinical use in spinal cord injury (SCI). Silk fibroin (SF)-based hydrogels have emerged as a promising strategy for local, controlled, and sustained drug release in the treatment of various diseases. In this study, we developed a SF-based hydrogel for the sustained delivery of GM1 and bMSCs (GM1 + bMSCs@SF hydrogel) and evaluated its therapeutic effect in a rat model of SCI. The GM1 + bMSCs@SF hydrogel showed sustained GM1 release in vitro. Treatment with the GM1 + bMSCs@SF hydrogel increased cell cycle exit and neuronal differentiation 1 (Cend1) expression, reduced neuronal pyroptosis and apoptosis, and promoted the differentiation of bMSCs into neurons. Double immunofluorescence staining confirmed a decrease in cell pyroptosis. Moreover, rats treated with the GM1 + bMSCs@SF hydrogel showed significant improvement in motor function recovery, with the Basso, Beattie and Bresnahan (BBB) score at 8 weeks reaching about 71.4% of the normal level, compared with about 30% in the SCI group. This study will minimize neuronal damage by innovatively using SF hydrogel loaded with GM1 and bMSCs for spinal cord injury repair. It will also explore the molecular mechanisms underlying post-traumatic spinal cord injury repair, opening new therapeutic avenues for spinal cord injury treatment.
GW231123, the heaviest binary black hole merger detected by the LIGO-Virgo-KAGRA Collaboration to date, lies in the pair-instability mass gap and exhibits unusually high component spins. In this Letter, we show that both merging black holes may have a primordial origin with smaller initial masses. The observed masses and, crucially, the spins of GW231123 are naturally accommodated within the most vanilla primordial black hole framework, once cosmological accretion is taken into account. Interestingly, the parameter space needed to explain the inferred GW231123 rate is at the edge of the exclusion region from x-ray and CMB observations, suggesting that this interpretation can be either confirmed or ruled out. The upcoming O5 observing run by the collaboration should detect O(20) similar events, testing their mass-spin correlation, while next-generation detectors would be capable of observing high-redshift events, as predicted in this scenario.
Natural killer (NK) cell-derived exosomes represent promising cell-free immunotherapeutics, yet clinical translation is limited by low production yield and lack of scalable manufacturing strategies. Here, we developed a modified vertical-wheel bioreactor incorporating controlled electrical and hypoxia-mimetic conditioning to enhance NK-exosome production while preserving biological functionality. The Electrical and Chemical Hypoxia-Optimized (ECHO) Bioreactor increased exosome yield by 10.1 ± 0.4 -fold compared to conventional culture (n = 3), without altering particle size, zeta potential, or canonical exosomal marker expression. In vitro, NK-Exo and NK-Exo-CE reduced LLC cell viability to 37.8 ± 3.1% and 18.6 ± 2.4%, respectively, at 1 × 10¹¹ particles/mL (p < 0.001). Combination therapy with paclitaxel further reduced viability to 9.4 ± 1.8% (p < 0.001). In a subcutaneous LLC tumor model, NK-Exo and NK-Exo-CE reduced tumor volumes to 912 ± 104 mm³ and 824 ± 96 mm³, respectively, compared to 1187 ± 132 mm³ in controls (p < 0.001), while NK-Exo-CE + PTX achieved 348 ± 57 mm³ (~ 3.4-fold reduction vs. control; p < 0.001) at the end of day 16. Immune profiling as determined by flow cytometry demonstrated increased CD8⁺ T cell infiltration (8.9 ± 1.1% to 35.7 ± 2.6%, p < 0.001) and reduced regulatory T cells (28.7 ± 1.8% to 10.4 ± 1.2%, p < 0.001) in combination-treated tumors. Western blot analysis confirmed elevated cleaved caspase-3, cleaved PARP, Granzyme B, FasL, Phospho-NF-κB-p65 and reduced Ki-67 and PDL1 in NK-Exo-CE + PTX tumors. Collectively, electro-hypoxic bioreactor conditioning enables scalable NK-exosome production without compromising physicochemical integrity or antitumor efficacy, addressing a key translational barrier in extracellular vesicle therapeutics.
Acorns are nutrient-rich fruits in which starch and proanthocyanidins (PAs) constitute the two dominant storage and defense metabolites. Transcriptomic studies in oaks have focused on stress responses, cork formation, and somatic embryogenesis, while omics and metabolic studies of acorn development remain scarce. We hypothesized that starch and PA biosynthesis in Quercus glauca fruits are governed by stage-specific transcriptional programs that drive their differential accumulation during fruit maturation. In this study, we utilized transcriptome data to investigate the expression dynamics of genes associated with starch and PA biosynthesis during Quercus glauca fruit development. Starch content increased continuously from 5.0 to 9.5 mg/mg protein across the five developmental stages of Q. glauca fruit, whereas PA content fluctuated within 0.015-0.038 mg/mg protein, peaking at T3 and T5. Transcriptome sequencing identified 2,637 differentially expressed genes (DEGs), predominantly partitioned into the rapid fruit growth phase (T3-T5) and enriched in PA biosynthesis, sugar metabolism, seed germination, and cell-cycle processes. Weighted gene co-expression network analysis (WGCNA) identified five modules significantly correlated with starch or PA content, within which 50 hub genes including QgPAL7 were identified. Mfuzz clustering showed PA biosynthesis genes were induced early and progressively repressed toward maturation, whereas starch biosynthesis genes were continuously up-regulated. WGCNA co-expression and metabolite-correlation analyses identified QgMYB33 as a stage-specific candidate associated with both PA and starch biosynthesis. The GPT family exhibited expansion through both tandem and segmental duplications. RT-qPCR validation of nine selected genes confirmed the RNA-seq expression patterns. This study characterizes a biphasic transcriptional program coordinating starch and PA biosynthesis during Q. glauca fruit development and identifies QgGPT7, QgGPT10, QgPAL7 and QgMYB33 as candidates, providing a molecular foundation for the future utilization and improvement of acorn resources.
Photosynthesis, which sustains life on Earth, depends on organized and yet adaptable protein assemblies embedded in specialized membranes known as thylakoids. Understanding how these complexes interact and reorganize within functional photosynthetic membranes is essential to reveal the molecular basis of energy conversion in cells. Here, we present an improved cross-linking mass spectrometry strategy that captures native protein interactions in photosynthetically active thylakoid membranes from Arabidopsis thaliana and Spinacia oleracea. By monitoring photosynthetic performance during cross-linking, we show that electron transport remains active, allowing structural analysis under physiological conditions. Moreover, we show that trimethylphenylammonium chloride as an adjuvant charged compound does not impair physiological activity, while boosting and diversifying cross-link identifications. Mapping cross-links onto known structures confirms the integrity of major photosynthetic complexes and uncovers previously uncharacterized assemblies involving regulatory and structural proteins. Integration with structural modeling and interaction network analysis identifies novel protein players within the photosynthetic machinery, providing molecular insights into their potential roles. This approach offers a broadly applicable framework for studying membrane protein organization and dynamics in functional bioenergetic systems.
Self-harm is a frequent feature of borderline personality disorder. In severe conditions comorbidity across mental disorders is common. Intellectual developmental disorder (IDD) may be an underlying condition with high frequent self-harming behavior. Based on extensively hospitalized severe self-harm patients, we aimed to 1) explore associations between possible IDD (identified by Hayes Ability Screening Index, HASI), and mental health disorders, and 2) investigate the ability to identify possible IDD with a low-effort HASI subscale. We hypothesized that possible IDD was significantly associated with greater mental health comorbidity. The study included 42 adults admitted to psychiatric hospitals for severe self-harm. Assessments included diagnostic interviews and IDD screening (HASI) by clinicians, and childhood trauma (patient self-report). Data were analyzed using Poisson regression. Validation of the HASI subscale background information was conducted using ROC analysis and exploratory data analysis. Possible IDD was significantly associated with higher incidence of mental disorders, even after adjusting for BPD, SUD, autism, and childhood trauma. Possible IDD and childhood trauma were independently associated with the number of mental disorders. The HASI background subscale demonstrated good predictive properties for identifying possible IDD. Results highlight the importance of assessing IDD among severe self-harm inpatients. A diagnosis of IDD may open the door to a broader range of treatment adaptations specifically addressing the needs of individuals with complex developmental and psychological challenges. Given the small sample size, these findings should be viewed as preliminary and hypothesis-generating. Replication in larger samples is required for further confirmation.
Familial hypercholesterolaemia (FH) is a common but underdiagnosed genetic condition that leads to high low-density lipoprotein cholesterol and premature cardiovascular disease. Cascade genetic testing remains underutilised as a screening approach. Implementation strategies can increase the diffusion of innovations such as genetic medicine into non-genetic specialty settings more rapidly through dissemination networks. This study aimed to evaluate the success of a multifaceted implementation strategy to increase cascade testing for relatives of a person diagnosed with FH using a primary-tertiary shared care model. A multisite effectiveness-implementation hybrid type III pre-post study was conducted between 2022 and 2024 to compare the implementation of a new shared care model for FH cascade testing in NSW, Australia. During the control period, cascade testing of relatives was offered to FH index cases by a genetic counsellor at the lipid clinic. In the implementation strategy period, FH index cases were offered a cascade testing expression of interest form to provide to their relatives. Relatives were contacted by a genetic counsellor and provided with a pre- and post-cascade testing package, which was also provided to their general practitioner. A total of n = 25 index and n = 6 cascade cases were included in the control period and n = 81 index and n = 56 cascade cases in the implementation period. The number of cascade genetic tests per index case increased significantly from 0.24 in the control period to 0.52 in the implementation strategy period (incident rate ratio = 4.62; 95% CI: 0.39, 8.84; p = 0.032). Among relatives tested, there was no difference in the proportion with a confirmed FH gene change per index case (IRR = 2.15; 95% CI: 0.65, 7.01; p = 0.207). There was no difference in the proportion of index cases with at least one cascade test (OR = 1.49; 95% CI: 0.53, 4.19; p = 0.442). Implementation of our model increased FH cascade testing compared to previous standard care. These findings suggest a greater role for cascade testing in primary care settings. This opens new opportunities for integrating genetic screening into routine general practice, particularly for autosomal dominant conditions such as FH.