Search Results (293)

Understanding the reproductive biology of orchids is essential for evaluating population viability and guiding conservation strategies, as their persistence often depends on complex interactions between ecological, physiological, and environmental factors. Govenia capitata, a threatened orchid endemic to the montane forests of central Mexico, had not previously been studied in this regard. We examined flowering phenology, floral longevity, stigmatic receptivity, natural and experimental pollination success, seed viability, and asymbiotic germination in two wild populations. Flowering was synchronous, with inflorescences lasting up to 57 days and individual flowers persisting for an average of 20 days. Stigmatic receptivity was detectable from the first day of anthesis and remained evident for at least eight days. Natural fruit set was very low (16.6%), while assisted self- and cross-pollination reached 100% success, demonstrating self-compatibility despite the inability for autonomous selfing due to floral structure. Seed viability differed significantly among treatments, being lowest in selfed capsules (11%) and highest in cross-pollinated ones (32%), representing a 65% reduction and reflecting severe inbreeding depression that extended to germination performance. In vitro germination success also varied, with the L-arginine medium yielding the highest values (46% for cross-pollinated seeds and 44% for naturally pollinated seeds), though post-germination survival requires optimization for conservation applications. Despite the conspicuous floral display, floral visitation was extremely rare and the pollinator identity remains unknown, with only one potentially effective visitor observed during 144 h of monitoring, and most floral visitors were non-pollinating arthropods such as crab spiders, weevils, hymenopterans, and thrips. Population density varied dramatically (26-fold) between sites separated by less than 1 km, indicating pronounced sensitivity to local environmental conditions. These findings reveal that reproduction in G. capitata is constrained by both extrinsic (pollinator limitation) and intrinsic factors (reduced seed viability), which collectively jeopardize long-term population persistence. From a conservation perspective, protecting montane forest remnants and pollinator communities is essential, while the demonstrated potential of asymbiotic germination provides a complementary tool for ex situ propagation and management of this endemic orchid. Full article

Chronic hemophilic synovitis (CHS), driven by hemosiderin-laden macrophages from recurrent hemarthrosis, is a major cause of joint damage in hemophilia. Platelet-rich plasma (PRP) is a promising regenerative therapy for joint diseases. This study investigated PRP’s ability to modulate macrophage polarization from a pro-inflammatory (M1) to a pro-resolving, tissue-repairing (M2) phenotype in CHS. We analyzed synovial fluid (SF) from CHS patients (N = 22), both pre- and post-PRP treatment. Ex vivo analysis revealed a predominant M1 profile with an increased proportion of CD11⁺CD14⁺CD64^hi compared with CD206⁺ or CD163⁺ M2 macrophages in CHS SF. In vitro experiments showed that CHS SF skewed monocyte-derived macrophages toward an M1 inflammatory program, evaluated by flow cytometry, qPCR, and ELISA. However, adding PRP significantly modulated the pro-inflammatory macrophage program, promoting an M2 tissue repair profile. Furthermore, a random forest machine learning algorithm, applied to public scRNAseq data, confirmed PRP’s macrophage reprogramming effect. Functional assays also showed increased TGF-β secretion and macrophage fusion when challenged with neutrophil extracellular traps (NETs). A small patient follow-up cohort treated with intra-articular PRP showed similar results, including normalization of cellular content and reduced CD64/CD206 expression. These findings indicate that PRP treatment effectively shifts SF-associated M1 macrophages to an M2-like phenotype, highlighting its potential as a therapeutic strategy for CHS. Full article

Chronic thromboembolic pulmonary disease (CTEPD) with or without pulmonary hypertension (PH) is characterized by persistent perfusion defects and progressive pulmonary vascular dysfunction after acute pulmonary embolism (PE), despite adequate anticoagulant therapy. We aimed at assessing clinical, hemodynamic, and functional evolution in patients screened for CTEPD with persistent lung perfusion scintigraphy (Q-scan) defects to identify non-invasive predictors of right ventricular (RV) impairment and development of exercise-induced pulmonary hypertension (ExPH). We analyzed 55 patients with a history of PE and no prior cardiopulmonary disease, stratified by perfusion (Q)-scan at 4 months into Q-scan-positive (n = 35) and Q-scan-negative (n = 20). At that time, all patients underwent echocardiography, cardiopulmonary exercise testing (CPET), and exercise stress echocardiography (ESE). Clinical evaluation and resting echocardiography were repeated at 24, 36, and 48 months. At baseline, Q-positive patients had higher NT-proBNP levels and greater PESI scores. At 4 months, they exhibited a higher prevalence of exercise-induced pulmonary hypertension (ExPH) on both CPET and ESE (p < 0.001). Both groups showed a partial recovery of echocardiographic parameters over time; however, Q-positive patients featured significantly higher systolic (s) pulmonary artery pressure (PAP) and mean PAP and a lower TAPSE/sPAP ratio (p < 0.001), increased eccentricity index, and shorter right ventricle (RV) outflow tract acceleration time at 48 months, suggestive of persistent RV-PA uncoupling and of a higher subclinical hemodynamic burden. Persistent Q-scan defects identify a post-PE population at risk for long-term RV dysfunction and ExPH, even in the absence of pulmonary hypertension at rest. CPET and ESE at 4 months provide useful prognostic information, supporting their integration into structured follow-up strategies to identify patients early on with evolving pulmonary vascular disease. Full article

Background and Objectives: The treatment of Chronic Myeloid Leukemia (CML) faces challenges such as resistance to Tyrosine Kinase Inhibitors (TKIs), necessitating new adjuvant therapies. This study aimed to evaluate the cytotoxic effect of direct, photosensitizer-free irradiation with LASER and LED light on the CML cell line K-562, hypothesizing that LASER light at a specific wavelength would be selectively effective. This work serves as a foundational in vitro study to establish the basis for a potential ex vivo therapeutic strategy. Methods: The human CML cell line K-562 was irradiated with LASER (405, 532, 629 nm) and LED (457, 517, 630 nm) sources at energy doses from 1 to 10 J/cm². Cell viability was assessed 24 h post-irradiation using Trypan Blue exclusion, the MTT assay, and biophysical changes in the cell absorbance spectrum. Results: Irradiation with a 532 nm LASER was the only condition that induced massive, statistically significant, and dose-dependent cytotoxicity, reaching up to 67.8% cell death at 10 J/cm² (p < 0.05). In contrast, other LASER wavelengths and all tested LED wavelengths failed to produce a significant cytotoxic effect. The superiority of the LASER over the LED of a similar wavelength highlights the critical role of the physical properties of light. Conclusions: Direct, photosensitizer-free irradiation with 532 nm LASER light is a potent and selective method for inducing cytotoxicity in K-562 cells in vitro. This effect is critically dependent on both the specific wavelength and the optical properties of the light source. These findings establish a solid foundation for the development of new ex vivo adjuvant therapies, such as extracorporeal photopheresis, for CML, pending further validation of its mechanism and selectivity. Full article

Background/Objectives: This exploratory study aimed to evaluate the impact of an 80 km ultra-marathon trail running event on changes in faecal bacterial composition, and to investigate whether any correlations exist between exercise-associated gastrointestinal symptoms (Ex-GIS) with faecal bacterial profiles. Such events represent a unique physiological stressor and may impact the composition of the gut microbiota. Studying this impact may provide insights into acute (i.e., <24 h) gut microbiota changes under extreme conditions. Methods: Thirteen endurance athletes (n = 7 males, n = 6 females) aged 41 ± 8 years completed the 80 km Margaret River (Australia) ultra-marathon race in 2022. Faecal samples were collected pre- and post-race. Faecal bacterial profile, as per relative abundance (RA) of operational taxonomic units and the determination of α-diversity (Shannon Equitability Index (SEI)), was achieved by 16S rRNA amplicon gene sequencing. Changes in RA% and SEI pre- to post-race were assessed by the Wilcoxon signed-rank test. Correlations between Ex-GIS with bacterial profile and changes pre-, during, and post-ultra-marathon race were determined by Spearman’s rank correlation coefficients. Results: Bacterial calculations of phyla (n = 5), family (n = 23), and genus (n = 41) were detected for RA (≥0.5%). A significant decrease pre- to post-race of Actinobacteriota (p = 0.035) phyla, Bifidobacteriaceae (p = 0.007), and Clostridiaceae (p = 0.010) family, and Blautia (p = 0.039) and Subdoligranulum (p = 0.023) genus was determined; meanwhile, Oscillospiraceae (p = 0.016) and Monoglobaceae (p = 0.039) family significantly increased pre- to post-race. No other bacterial group changes were observed. No correlations were observed between pre- to post-ultra-marathon RA change and Ex-GIS. Conclusions: The completion of an 80 km ultra-marathon did not invoke substantial changes in the gut microbiota as determined by faecal bacterial profiling. Very strong and strong correlations were observed between certain bacterial groups and Ex-GIS; however, no significant correlations were observed between pre- to post-ultra-marathon changes in RA ≥ 0.5% and Ex-GIS. Full article

Global amphibian populations are declining rapidly and the development of effective cryopreservation protocols for germ cells has become a critical tool in ex situ conservation programs. Post-thaw membrane-integrity viability in four endangered Korean amphibians (Dryophytes suweonensis, Pelophylax chosenicus, Kaloula borealis, and Hynobius yangi) were evaluated. Sperm were cryopreserved using dimethyl sulfoxide (DMSO) or N,N-dimethylformamide (DMF) at 10–30% (v/v) in combination with 0.6 M sucrose, and were frozen at two suspension heights (5 cm vs. 10 cm) above liquid nitrogen. Post-thaw membrane-integrity viability was assessed using a SYBR-14/propidium iodide membrane-integrity assay (LIVE/DEAD kit). Low concentrations of permeating cryoprotectants (CPs) improved membrane-integrity viability, whereas high concentrations led to high toxicity, particularly with DMSO. Across species, DMF produced the highest membrane-integrity viability and the most consistent performance. The cooling rate influenced membrane-integrity viability, with samples frozen at 10 cm exhibiting greater viability, reflecting the balance between intracellular ice formation during rapid cooling and solution effects during slow cooling. Optimal conditions for D. suweonensis were 15% DMSO at 10 cm (86.5% membrane-integrity viability); for P. chosenicus, 10% DMF at 10 cm (75.5%); and for K. borealis, 10% DMSO at 5 cm (81.6% membrane-integrity viability). Hynobius yangi showed modest improvement under 15% DMF at 5 cm (19.7%), although overall membrane-integrity viability was low. ED₅₀ modeling indicated species-specific thresholds requiring low CP concentrations. Sperm cryopreservation outcomes in amphibians are strongly influenced by CP type, concentration, cooling regime, and species physiology. GLM and ED₅₀ modeling provide a methodological framework for refining cryopreservation strategies for non-model, endangered species. Full article

Background: Common variable immunodeficiency (CVID), the most prevalent symptomatic inborn error of immunity, involves impaired B-cell differentiation and antibody production, causing recurrent infections and the need for life-long immunoglobulin replacement therapy. Methods: This study evaluated the in vitro differentiation of memory B-cells (MBCs) into antibody-secreting cells (ASCs) in CVID patients. Peripheral blood mononuclear cells from 13 CVID patients and 10 healthy controls were stimulated using two protocols: (I) Staphylococcus aureus Cowan Strain I, Pokeweed mitogen, and CpG, or (II) a T-cell-dependent approach using CD40 ligand, interleukin-21, and CpG. B-cell subpopulations were analyzed by flow cytometry, ASC differentiation using ELISpot, and antibody levels in supernatants by ELISA. Results: Despite severely restricted in vivo antibody production, MBCs from all 13 CVID patients differentiated into IgG and IgM ASCs under adequate in vitro stimulation. Protocol II, mimicking T-cell help, was more effective than protocol I. As expected, the patients exhibited reduced class-switched MBCs ex vivo, but the MBCs differentiated and proliferated to an extent similar to those in healthy controls. IgA secretion remained significantly impaired post-stimulation. Specific IgG antibodies against tetanus toxoid and Streptococcus pneumoniae were detected in the patient supernatants, while no double-stranded DNA autoantibodies emerged after in vitro stimulation. Conclusions: These findings indicate that the MBCs of most patients retain functional B-cell differentiation and antigen-specific IgG secretion under T-cell dependent stimulation, though IgA secretion remains impaired. Tailored stimulation protocols could deepen our understanding of how to restore MBC formation in CVID patients in vivo. This methodology provides a platform to investigate antigen-specific functional memory responses like post-vaccination. Full article

The 2020 Europe Strategy was designed as a comprehensive framework to promote smart, sustainable and inclusive growth in the European Union (EU), particularly emphasising the ‘20/20/20’ targets related to climate protection and energy policy. This study provides an ex post evaluation of the extent to which the strategy’s objectives were achieved in the member states of the EU in the period 2010–2020. The analysis is based on Eurostat data and uses Hellwig’s multidimensional comparative analysis to construct a synthetic indicator of progress. The results show that EU countries have made significant advances in reducing greenhouse gas emissions and increasing the share of renewable energy in gross final energy consumption, with Sweden and Finland identified as leaders, while Malta and Hungary lagged behind. Primary energy consumption overall decreased, although only a minority of the member states reached the planned thresholds. Progress was less evident in research and development (R&D) expenditure, where the average value of the EU remained below the 3% GDP target, and strong disparities persisted between innovation leaders and weaker performers. Improvements in higher education attainment were observed, contributing to the long-term goal of a knowledge-based economy, although labour market difficulties, especially among young people, remained unresolved. The findings suggest that, although the Strategy contributed to tangible progress in several areas, uneven achievements among member states limited its overall effectiveness. The study is limited by the reliance on aggregate statistical data and a single methodological approach. Future research should extend the analysis to longer time horizons, include qualitative assessments of national policies, and address implications for the implementation of the European Green Deal and subsequent EU development strategies. Full article

Background: Hip fractures are common in the elderly and often require ICU admission post-surgery due to high ASA scores and comorbidities. Length of hospital stay after ICU is a crucial indicator affecting patient recovery, complication rates, and healthcare costs. This study aimed to develop and validate a machine learning-based model to predict the factors associated with extended hospital stay (>7 days from surgery to discharge) in hip fracture patients requiring postoperative ICU care. The findings could help clinicians optimize ICU bed utilization and improve patient management strategies. Methods: In this retrospective single-centre cohort study conducted in a tertiary ICU in Turkey (2017–2024), 366 ICU-admitted hip fracture patients were analysed. Conventional statistical analyses were performed using SPSS 29, including Mann–Whitney U and chi-squared tests. To identify independent predictors associated with extended hospital stay, Least Absolute Shrinkage and Selection Operator (LASSO) regression was applied for variable selection, followed by multivariate binary logistic regression analysis. In addition, machine learning models (binary logistic regression, random forest (RF), extreme gradient boosting (XGBoost) and decision tree (DT)) were trained to predict the likelihood of extended hospital stay, defined as the total number of days from the date of surgery until hospital discharge, including both ICU and subsequent ward stay. Model performance was evaluated using AUROC, F1 score, accuracy, precision, recall, and Brier score. SHAP (SHapley Additive exPlanations) values were used to interpret feature contributions in the XGBoost model. Results: The XGBoost model showed the best performance, except for precision. The XGBoost model gave an AUROC of 0.80, precision of 0.67, recall of 0.92, F1 score of 0.78, accuracy of 0.71 and Brier score of 0.18. According to SHAP analysis, time from fracture to surgery, hypoalbuminaemia and ASA score were the variables that most affected the length of stay of hospitalisation. Conclusions: The developed machine learning model successfully classified hip fracture patients into short and extended hospital stay groups following postoperative intensive care. This classification model has the potential to aid in patient flow management, resource allocation, and clinical decision support. External validation will further strengthen its applicability across different settings. Full article

Antecedents: From a functional perspective, executive functions—such as self-regulation and meta-cognition—emerge as key dimensions affected transversally across various neurodevelopmental disorders. Aim: The aim of this study is to analyze and compare executive functioning profiles in children with various neurodevelopmental disorders, as reported by parents and teachers. It is hypothesized that children with neurodevelopmental disorders exhibit executive function deficits, as measured by the BRIEF-P, in comparison to typically developing children. Methodology: We used a non-experimental methodology and ex post facto design to perform a descriptive, cross-sectional study. Participants: The normative sample is composed of 1.979 participants with typical (normotypical) development and 205 participants belonging to a clinical sample. Measurement: The instrumental development of EFs was evaluated using BRIEF-P by key informants. Results: The highest F-values were observed in the following: (i) working memory, (a) parents [F = 195.76, p < 0.001] and (b) teachers: [F = 199.63, p < 0.001]; and (ii) Emergent Metacognition Index, (a) parents [F = 176.15, p < 0.001] and (b) teachers [F = 187.87, p < 0.001]; and (iii) Executive Function Global, (a) parents [F = 168.07, p < 0.001] and (b) teachers [F = 207.47, p < 0.001]. Conclusions: This study provides a clear framework for identifying dysexecutive syndrome. Executive functioning is one of the most important abilities, and its disruption can lead to dysexecutive syndrome. Full article

This study aims to investigate a fundamental yet largely overlooked question: “Does investing in transportation infrastructure positively impact farms’ agricultural gross income?” It is examined based on the role of transportation infrastructure in ensuring equal access to market opportunities in the context of the widening regional economic disparity in Korea. The main novelty of this study lies in its attempt to introduce an accessibility measure for evaluating the benefits of transportation infrastructure in a rural setting, which has been limitedly applied in urban-centered studies. To accomplish this task, multilevel and spatial econometric models were employed to evaluate the ex-post impact of transportation accessibility on agricultural gross income from the perspectives of farmers, primarily, and rural autonomies, subsequently. This study found that the continuation of the current direction of transportation policy—without substantial consideration for agriculture as an industry and rural areas as living spaces—can intensify the economic alienation of agriculture and rural areas. This study concludes that opportunities for market access provided by the immense public investments in transportation infrastructure should be fairly distributed to farmers and rural autonomies to promote balanced regional development in Korea. Full article

The objective of this study was to evaluate the effect of fire on the recruitment of six woody species sown at two soil depths. The study area was the western Argentine Chaco region, South America. The selected species were Aspidosperma quebracho-blanco Schlecht, Schinopsis lorentzii (Griseb.) Engl., Sarcomphalus mistol (Griseb.) Hauenschild, Neltuma nigra (Griseb.) Hieron, Senegalia gilliesii (Steud.) Seigler & Ebinger, and Vachellia aroma (Gillies ex Hook. & Arn.) Seigler & Ebinger. Seedling emergence was monitored following experimental burns (EB) in 2017 and 2019 under similar environmental conditions. In 2017, seeds were placed on the litter layer in 2 × 2 m plots; in 2019, seeds were buried at 2 cm depth in 2.5 × 10 m plots. Field emergence was recorded, and viability of non-germinated seeds was tested in the lab. Recruitment was also assessed two years post-EB 2017 in 20 plots (2 × 5 m) under burnt and control conditions. No emergence occurred from seeds on the litter layer, but S. lorentzii and N. nigra germinated in the laboratory (<7%, significant difference, p < 0.0001). All species with buried seeds germinated in the field, with significant differences among species and compared to controls (p < 0.0027). Field recruitment revealed regeneration of A. quebracho-blanco, N. nigra, and S. gilliesii in the burnt area, with variations in seedling size and density. Full article

Objectives: We investigated the in vivo biodistribution of vascular cell adhesion molecule-1 (VCAM-1)-targeting polystyrene nanoparticles (PS-NPs) labeled with Rhodamine B in a murine model of atherosclerosis. Methods: Targeted PS-NPs of varying sizes were first assessed for in vitro uptake in RAW264.7 cells. In vivo evaluation with VCAM-1-targeted nanoparticles (NP T) in C57 BL/6NtaC mice was conducted, and organs were analyzed 1, 6, and 24 h post injection, ex vivo. Subsequently, both targeted (NP T) and non-targeted (NP NT) nanoparticles of 30, 60, and 120 nm were injected into Apolipoprotein E knock-out (ApoE KO) mice on a high-fat diet, with ex vivo organ analysis 24 h post injection. Results: Results showed that NP30 T and NP60 T accumulated primarily in the liver and kidney of B6 mice. In ApoE KO mice, biodistribution was largely unaffected by size and targeting, except for higher uptake of NP 120 NT and T in the lungs and spleen. All NP types, except NP60 NT, showed significantly higher signal in ApoE KO mouse aortas compared to saline controls, with no significant differences between NP groups. Conclusions: While nanoparticles accumulated significantly in ApoE KO mouse aortas compared to controls, size and targeting properties did not significantly affect biodistribution in major organs 24 h post injection. Full article

(1) Background: The present study aimed to evaluate the effects of fixed orthodontic treatment using the Damon self-ligating system on incisor inclination and soft tissue facial profile. Additionally, the study compared the outcomes of premolar extraction versus non-extraction approaches on these parameters. (2) Methods: This retrospective study included 72 patients who were treated with the Damon self-ligating bracket system (Damon, Ormco, Orange, CA, USA). Thirty-five patients (mean age 12.5 years; 20 girls and 15 boys) underwent premolar extraction (Group Ex), while 37 patients (mean age 12.8 years; 16 girls and 21 boys) were treated without extractions (Group NonEx). Lateral cephalometric radiographs were taken pre- and posttherapeutically, and measurements were conducted using OnyxCeph3™ (Image Instruments, Chemnitz, Germany). (3) Results: Following orthodontic treatment, the upper incisors in group Ex were significantly more retruded (p < 0.01), while the mandibular incisors in group NonEx were significantly more proinclined (p < 0.01). Both lips in group Ex, and the upper lip in the group NonEx, exhibited a greater distance from the esthetic line post-therapy (p < 0.01). The position of the lips relative to the SR line did not significantly change in group Ex, whereas group NonEx showed a significantly increased lip protrusion (p < 0.01). A weak negative correlation (−0.37) was observed between the increased retroclination of the upper incisors and the retrusion of the upper lip relative to the SR line. (4) Conclusions: Premolar extraction, as part of a treatment strategy for cases with severe crowding, combined with Damon™ self-ligating braces, does not adversely affect the soft tissue profile. Full article

Objective: The objective of this study was to identify potential robust acoustic biomarkers for functional post-thyroidectomy voice disorder (PTVD) that may support early diagnosis and personalized treatment strategies, using acoustic analysis and explainable machine learning methods. Methods: Spectral and cepstral features were extracted from /a/ and /i/ voice recordings collected preoperatively and 4–6 weeks postoperatively from a total of 126 patients. Various Support Vector Machine (SVM) and Boosting models were trained. SHapley Additive exPlanations (SHAP) analysis was applied to enhance interpretability. SHAP values from training and test sets were compared via scatter plots to identify stable candidate biomarkers with high consistency. Results: GentleBoost (AUC = 0.85) and LogitBoost (AUC = 0.81) demonstrated the highest classification performance. Performance metrics across all models were evaluated for statistical significance. DeLong’s test was conducted to assess differences between ROC curves. The features iCPP, aCPP, and aHNR were identified as stable candidate biomarkers, exhibiting consistent SHAP distributions in both training and test sets in terms of direction and magnitude. These features showed statistically significant correlations with PTVD (p < 0.05) and demonstrated strong effect sizes (Cohen’s d = −2.95, −1.13, −0.60). Their diagnostic relevance was further supported by post hoc power analyses (iCPP: 1.00; aCPP: 0.998). Conclusions: SHAP-supported machine learning models offer an objective and clinically meaningful approach for evaluating PTVD. The identified features may serve as potential biomarkers to guide individualized voice therapy decisions during the early postoperative period. Full article