Search Results (50,461)

Background/Objectives: Urine samples are the most frequently analyzed specimens in clinical microbiology laboratories. Although urine culture remains the gold standard for diagnosing urinary tract infections, it is time-consuming and resource-intensive. Therefore, reliable screening methods capable of predicting urine culture positivity are needed to optimize laboratory workflow. Automated urine analysis based on flow cytometry enables efficient screening and identification of samples with a low probability of bacterial infection, thereby rationalizing microbiological testing. This study evaluated the usefulness of a multivariable approach to support interpretation of flow cytometry results following the implementation of the Sysmex UF-4000 urine flow cytometer. Methods: Routinely collected urine samples from outpatients and hospitalized patients were analyzed using the UF-4000 flow cytometer, with a positivity threshold of ≥100 leukocytes/µL. Urinary parameters were compared between samples with positive and negative cultures. Multivariable logistic regression was applied to identify independent predictors of a positive urine culture. Urinary sediment parameters, including leukocyte, bacterial, fungal, and squamous epithelial cell counts, were assessed as covariates. Results: Urine samples with positive cultures showed significantly higher leukocyte counts (median 355.0, IQR 146.5–1429.4) and bacterial counts (median 9805.2, IQR 1134.3–45,011.5). Fungal and squamous epithelial cell counts differed only slightly between groups, although the differences were statistically significant (p < 0.001). Leukocyte counts were higher in urine samples from which Gram-negative bacteria were isolated compared with samples containing Gram-positive bacterial isolates (p < 0.001). The multivariable model demonstrated the most favorable overall performance, combining high sensitivity with improved specificity and the highest negative predictive value (AUC = 0.927). Optimal cut-off values were 70 leukocytes/µL and 105 bacteria/µL. Conclusions: Leukocyte and bacterial counts were the strongest predictors of positive urine culture results. A multivariable model including only these two parameters demonstrated high diagnostic accuracy and may serve as a practical screening tool to identify urine samples with a low probability of bacterial infection. The implementation of this approach could support more efficient use of urine cultures and help optimize laboratory workflow. Full article

Background and Objectives: Severe COVID-19 frequently fulfills Sepsis-3 criteria and is characterized by thrombo-inflammation and endothelial injury. We evaluated whether a bedside endothelial activation index (EAI = D-dimer/fibrinogen) identifies biologically distinct phenotypes and relates to interleukin-6 (IL-6) response after therapeutic plasma exchange (TPE), and whether baseline IL-6 predicts a ≥50% IL-6 reduction. Methods: Retrospective single-center ICU cohort of adults with SARS-CoV-2 infection, sepsis-related organ dysfunction, and ≥1 TPE session (n = 51). Patients were stratified by median EAI (low vs. high). Outcomes included peri-procedural biomarker/physiology changes (post–baseline), IL-6 responder status (≥50% reduction), correlations with IL-6 reduction (%), and multivariable predictors of response. Results: Compared with low EAI (n = 25), high EAI (n = 26) had higher baseline D-dimer (6.2 vs. 2.2 µg/mL) and lower fibrinogen (2.9 vs. 7.1 g/L) (both p < 0.001). Low EAI showed larger CRP decreases (ΔCRP −84.0 vs. −2.3 mg/L; p = 0.001) and larger fibrinogen falls (Δ −3.1 vs. −0.4 g/L; p < 0.001), while high EAI had larger D-dimer decreases (Δ −2.5 vs. −0.6 µg/mL; p = 0.004) and a modest SOFA improvement (Δ −0.3 vs. +0.1; p = 0.026). IL-6 responders (n = 20) had higher baseline IL-6 than non-responders (365.2 vs. 47.1 pg/mL; p < 0.001). Baseline IL-6 independently predicted response (per doubling: OR 1.94, 95% CI 1.27–2.95; p = 0.002), while age reduced odds (OR 0.91/year, 95% CI 0.84–0.99; p = 0.032). IL-6 reduction correlated with ΔCRP (ρ = −0.41; p = 0.003) and ΔPaO₂/FiO₂ (ρ = 0.37; p = 0.01). Conclusions: EAI stratifies distinct thrombo-inflammatory patterns around TPE, while baseline IL-6 is the dominant predictor of achieving large IL-6 reductions. To emphasize the novelty and clarify the study objective, this exploratory analysis used a phenotype-stratified framework to test whether a simple bedside endothelial activation index could enrich biological response assessment to adjunctive TPE. The prespecified primary outcome was achievement of a ≥50% IL-6 reduction after completion of the TPE course; secondary outcomes included peri-procedural biomarker, oxygenation, SOFA, and ICU endpoints. Full article

Background/Objectives: High-grade endometrial cancer (EC) is associated with poor outcomes, particularly in populations with a high burden of aggressive histologies. There is a critical need for accessible biomarkers to improve prognostic assessment and guide clinical management. Methods: In this study, we evaluated the feasibility and clinical relevance of monitoring circulating tumor DNA (ctDNA) by tracking somatic TP53 mutations using a routine next-generation sequencing (NGS) assay already implemented in diagnostic practice. Results: Among 21 patients with high-grade EC carrying TP53 mutations in the primary tumor, ctDNA was detectable in over 75% during follow-up. Baseline ctDNA detection strongly correlated with advanced disease: none of the FIGO I tumors were ctDNA-positive at diagnosis, whereas 73% of FIGO > I tumors showed detectable ctDNA. Patients with ctDNA detected at baseline had significantly poorer outcomes, with a 2-year recurrence-free survival (RFS) of 18% versus 60% and a 2-year overall survival (OS) of 40% versus 78%. Longitudinal monitoring revealed that postoperative persistence or reappearance of ctDNA was consistently associated with disease progression, often preceding radiological relapse. Conversely, early ctDNA clearance (at M4–M8) was associated with more favorable clinical trajectories. Conclusions: These findings highlight the potential role of ctDNA as a real-time molecular marker of minimal residual disease and tumor dynamics. Our results demonstrate that TP53-based ctDNA tracking using a standard NGS panel is feasible, sensitive, and clinically informative in high-grade EC. This approach may contribute to improving prognostic stratification and enabling more personalized, responsive clinical management, particularly in high-risk populations. Full article

Background and Objectives: Coronavirus disease 2019 (COVID-19) is characterized by excessive inflammatory responses, including the so-called cytokine storm, which contributes substantially to morbidity and mortality in hospitalized patients. The vagus nerve, through the cholinergic anti-inflammatory pathway, represents a theoretically attractive therapeutic target for modulating systemic inflammation. Vagus nerve stimulation (VNS) has emerged as a potential adjunctive treatment for COVID-19, with several randomized controlled trials (RCTs) investigating its efficacy on inflammatory biomarkers and clinical outcomes. The quality of this evidence base has not been rigorously evaluated. This systematic review critically appraises all available RCT evidence for VNS in hospitalized COVID-19 patients. Materials and Methods: We systematically searched PubMed, Scopus, Cochrane (CENTRAL), and Web of Science from database inception to January 2026, for RCTs evaluating any form of VNS (invasive, non-invasive, cervical, or auricular) in hospitalized patients with confirmed acute COVID-19. Two reviewers independently screened titles, abstracts, and full texts according to pre-specified eligibility criteria. Risk of bias was assessed using the Cochrane Risk of Bias 2 (RoB 2) tool, with assessments initially performed using multiple artificial intelligence tools and subsequently validated by the authors in accordance with PRISMA 2020 guidelines. Given substantial heterogeneity and high risk of bias, narrative synthesis was performed rather than meta-analysis. Also, GRADE assessment was performed. Results: From 437 records identified, six RCTs comprising 221 patients met the inclusion criteria. Five trials (83%) were rated as high risk of bias, primarily due to inadequate blinding, substantial baseline imbalances, significant missing data and extensive multiple testing without statistical correction. The single double-blind trial with a credible sham control (Rangon et al.) found null results across all outcomes, including clinical progression, ICU transfer, and mortality, while the five “high” risk-of-bias trials generally reported positive findings on various inflammatory markers and clinical outcomes. One trial (Corrêa et al.) measured heart rate variability as a direct indicator of vagal activation and found no change despite claiming anti-inflammatory effects, contradicting the proposed mechanism of action. Significant cognitive findings from an interim analysis (Uehara et al., n = 21) disappeared in the larger completed trial (Corrêa et al., n = 52), providing empirical demonstration of false positive findings in small, underpowered studies. Conclusions: Currently available evidence supporting the use of VNS for acute COVID-19 remains scarce; however, the physiological rationale remains sound, although the absence of reliable target engagement markers in the included studies limits confidence in this treatment method. Large-scale, double-blind, sham-controlled trials are required before VNS can be firmly recommended for COVID-19 management. Full article

Background/Objectives: Routine liver biopsies play an important role in monitoring liver allografts but carry non-negligible risks. This pilot study assesses the feasibility of dynamic long-axial field-of-view (LAFOV) [¹⁸F]FDG PET/CT as a non-invasive alternative to biopsy. Methods: Liver transplant (LTx) recipients meeting the inclusion criteria of ≥10 months post-transplantation and scheduled routine biopsy were prospectively enrolled, along with healthy controls. All participants underwent dynamic LAFOV [¹⁸F]FDG PET/CT, followed by biopsy in LTx recipients, who were stratified by inflammatory severity using the BANFF score. Hepatic kinetic parameters (K1, k2, k3, k4) and SUVmean/SUVmax were compared using Mann–Whitney U tests. Correlations were assessed using Spearman’s rank correlation. A p-value < 0.05 was considered significant. Analyses were performed in RStudio (version 2024.12.10563). Results: Sixteen LTx recipients (mean age 48.6 years; seven female, nine male) and eight healthy controls (mean age 35.4 years; six female, two male) were included. Healthy controls had mean k3 and k4 values of 0.0037 min⁻¹ ± 0.0003 min⁻¹ and 0.0019 min⁻¹ ± 0.0011 min⁻¹, respectively. LTx recipients showed significantly higher k3 and k4 values, both when including and excluding patients with biopsy-confirmed inflammation. Descriptive comparisons between LTx recipients with and without significant inflammation (n = 3) showed no clear differences. Spearman analysis showed no significant correlations between the BANFF score and kinetic parameters. The strongest degree of correlation was found between BANFF score and k3, indicating a moderate positive but non-significant association (k3: rs = 0.396, p = 0.128). Conclusions: Elevated k3 and k4 values in LTx recipients were not explained by allograft inflammation, suggesting altered FDG kinetics post-transplant. These differences may confound [¹⁸F]FDG PET interpretation. Larger studies are needed to assess the clinical applicability of dynamic LAFOV [¹⁸F]FDG PET/CT. Full article

The objective of this study was to evaluate the impact of seasonal changes in the chemical and structural composition of giant miscanthus (Miscanthus × giganteus) biomass on the performance, kinetics, and efficiency of anaerobic digestion (AD), as well as on the overall energy and techno-economic balance of the conversion chain. The AD performance was assessed using batch biochemical methane potential (BMP) assays conducted for eight harvest dates (June–January). Comprehensive characterization included fundamental physicochemical properties of the biomass, lignocellulosic fraction composition, AD kinetics, and methane production yield. A statistically significant (p < 0.05) increase in structural fiber fractions was observed with advancing plant maturity, accompanied by a progressive decline in specific methane yield from 281 ± 32 mL CH₄/g VS in June to 170 ± 11–172 ± 13 mL CH₄/g VS in winter harvests. Despite a relatively stable theoretical biochemical methane potential (TBMP) ranging from 425 to 443 mL CH₄/g VS, the conversion efficiency (BMP/TBMP) decreased from approximately 66% to below 40%, indicating increasing structural and kinetic limitations to substrate biodegradability. Kinetic parameters deteriorated systematically in late harvests, as reflected by a reduction in the first-order rate constant k_CH₄ from 0.115 to approximately 0.072 1/d and an extension of the lag phase λ from 2.19 to over 4 days. Regression analysis revealed strong negative correlations between lignocellulosic complex content and both BMP and k_CH₄, whereas the C/N ratio exhibited a positive association with process performance under the experimental conditions applied. The highest methane production per hectare (3904 ± 720 m³CH₄/ha) and the most favorable economic outcome (1979 ± 465 EUR/ha) were achieved for the September harvest. The results demonstrate that harvest timing constitutes a critical optimization parameter in lignocellulosic biogas systems, governing not only methane yield and process kinetics but also the overall energy output and economic viability of the bioenergy production chain. Full article

Background/Objectives: This study aimed to examine the prevalence of anatomical variations in the sacroiliac joints (SIJs) as observed through Magnetic Resonance Imaging (MRI), to characterize their manifestations, and to identify MRI features that may resemble inflammatory alterations. Methods: A retrospective review was conducted on consecutive MRI scans of the SIJ performed from January 2009 to January 2022. Eight anatomical variations, along with associated edematous and structural changes, were assessed. Results: The study encompassed 840 patients, with anatomical variations identified in 39.7% of the cohort, occurring more frequently among female participants. The most prevalent variations were accessory SIJ (36.2%) and the iliosacral complex (32.2%). Notably, isolated synostosis and persistent ossification center variations were absent. The increased frequency of variations in women, as well as their correlation with advancing age, was statistically significant (p = 0.034). Accessory SIJ and dysmorphic alterations were linked to bone marrow edema and structural modifications. In the iliosacral complex and semicircular defect variations, prominent vascular structures were observed extending along the bone surfaces. The number and depth of edema slices in sacroiliitis exceeded those observed in the variation (p < 0.001). Conclusions: Anatomical variations of the SIJ are prevalent among women and tend to increase with advancing age. Given that these variations, particularly accessory SIJ and dysmorphic alterations, may present with edematous and structural signal intensity changes that resemble sacroiliitis, it is crucial to recognize these variations. It is recommended to assess axial and coronal images concurrently and to exercise caution in the interpretation of SIJ MR images. Full article

Objectives: The long-term effects of rheumatoid arthritis (RA) therapies on bone mineral density (BMD) remain incompletely characterized. We aimed to evaluate BMD trajectories over an 8-year follow-up in patients with RA treated with conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) or biological DMARDs (bDMARDs) in real-world practice. Methods: Patients were selected from an observational RA cohort established at Nice University Hospital between 2001 and 2016. Participants were classified into two groups according to treatment regimen (csDMARD only or any bDMARD exposure). BMD was assessed by dual-energy X-ray absorptiometry at baseline and after 1, 2, 3, 5, and 8 years at the lumbar spine, femoral neck, and total hip. Longitudinal changes in BMD were analyzed using multivariable linear mixed-effects models adjusted for age, sex, body mass index (BMI), disease duration, seropositivity, glucocorticoid use, anti-osteoporosis treatment, and clinical response. Results: A total of 312 patients were included, of whom 181 received bDMARDs and 131 were treated exclusively with csDMARDs. BMD showed limited change during the first two years in both groups. Beyond two years, modest declines were observed at hip sites at subsequent time points, whereas lumbar spine BMD did not demonstrate significant longitudinal change in pointwise analyses. In mixed-effects models, the treatment group–time interaction was significant for lumbar spine (p = 0.004) and total hip (p = 0.04), but not for the femoral neck (p = 0.34), indicating differential BMD trajectories over time between treatment groups. In the csDMARD group, lumbar spine and total hip BMD decreased by a mean of 0.0006 and 0.0005 g/cm² per month, respectively, whereas no significant slopes were observed in the bDMARD group. Older age was associated with lower BMD, while male sex and higher BMI were associated with higher BMD across sites. Conclusions: In this long-term real-world cohort, BMD remained relatively stable during the first two years of follow-up. Longitudinal analyses suggested a less pronounced decline in lumbar spine and total hip BMD trajectories among bDMARD-treated patients compared with those receiving csDMARD alone, underscoring the need for ongoing bone health monitoring in RA. Full article

Background/Objectives: Doxorubicin is an anthracycline chemotherapeutic agent widely used in the treatment of breast cancer. However, its clinical utility is limited by the drug’s resistance development, low oral bioavailability, and dose-dependent side effects. The semi-essential amino acid, L-arginine, has gained attention as a potential adjuvant that could improve the drug distribution and cytotoxic effectiveness of chemotherapeutics. This study aimed to explore the multifunctional effect of L-arginine on the intestinal absorption and anti-breast cancer activity of doxorubicin. Methods: The rabbit in situ intestinal perfusion technique was employed to investigate the membrane transport parameters of doxorubicin both in the absence and presence of L-arginine. Furthermore, the effect of L-arginine on the cytotoxic activity of doxorubicin against breast cancer cells (MCF-7) was assessed using the MTT assay. Results: Co-perfusion of L-arginine with doxorubicin enhanced the fraction of doxorubicin absorbed, with a recorded 4.3-fold enhancement in the jejuno-ileum and a 1.5-fold enhancement in the colon segment. In MCF-7 cells, co-treatment with L-arginine resulted in a significant potentiation of doxorubicin cytotoxicity. At L-arginine concentrations of 10 μM and 50 μM, the recorded IC₅₀ decreased from 41.3 μM to 8.2 μM and to 22.1 μM, respectively. The superior efficacy of 10 μM L-arginine compared to 50 μM reflected a biphasic concentration-dependent response. Conclusions: L-arginine modulated two critical aspects of doxorubicin efficacy, intestinal absorption and cytotoxic activity. The biphasic response emphasizes the importance of L-arginine dose optimization. These findings support the potential of L-arginine as a safe adjuvant for developing oral doxorubicin formulations. This approach can reduce the dose-related toxicity of doxorubicin and improve therapeutic outcomes. Full article

Background/Objectives: The Positive Parenting Program (Triple P) is an evidence-based intervention for families including young children with neurodevelopmental disorders. Because establishing a definitive diagnosis in early childhood takes time, especially for 1–3-year-olds who have only recently shown early signs, parents often experience substantial stress. This study examines the effectiveness of the Group Triple P (GTP) program for mothers of 1–3-year-old children without a confirmed diagnosis who spent time at a community parenting support center. Methods: The participants were 41 mothers of children aged 1–3 who did not confirmed neurodevelopmental diagnosis but showed behavioral, emotional, or developmental concerns, or whose mothers reported parenting difficulties. To reflect real community practices, a non-randomized pre–post test design without a control group was adopted. Assessments were conducted at baseline, postintervention, and at a 12 weeks follow-up using the Strengths and Difficulties Questionnaire, Parenting Scale, Parenting Experience Scale, and Depression Anxiety Stress Scale. Results: GTP improved the mothers’ assessments of positive behaviors in children who may have neurodevelopmental disorders, with a medium effect size. GTP reduced dysfunctional parenting styles in mothers with a large effect size. It also enhanced mothers’ confidence and fulfilling sense in parenting, and decreased depression and stress with a medium effect size. Perceived access to parenting support improved, contributing to better parenting adaptation with a medium effect size. Conclusion: GTP may serve as an early public health intervention for mothers of young children who may have neurodevelopmental disorders by supporting maternal mental health and promoting adaptive parenting. Full article

Objectives: As widely used first-line antihypertensive drugs, dihydropyridine calcium channel blockers (DHP-CCBs) have relatively few studies comparing their adverse reactions based on real-world data. This study aims to identify and compare the potential adverse drug reaction (ADR) signals of four DHP-CCBs (amlodipine, felodipine, nicardipine, and nifedipine) through the US Food and Drug Administration Adverse Event Reporting System (FAERS), providing a reference for further drug safety assessment and clinical medication risk awareness. Methods: Adverse event reports from medical professionals (Q3 2014–Q4 2024) were analyzed using signal detection methods, including reporting odds ratio (ROR), proportional reporting ratio (PRR), information component (IC), and the Medicines and Healthcare Products Regulatory Agency (MHRA) methods. Risk signals for the four DHP-CCBs were compared with both the full database and the DHP-CCBs background. For high-risk signals in amlodipine, multivariate logistic regression was used for validation. The analysis reveals distinct ADR profiles for the four DHP-CCBs. Results: Amlodipine is strongly linked to suicide-related risks, confirmed by logistic regression. Nicardipine and nifedipine show significant risks for pregnancy-related events, such as premature delivery and exposure during pregnancy. Nicardipine is also associated with hyponatremia, hyperkalemia, and lactic acidosis. These adverse events are not yet included in the FDA labeling for any of the DHP-CCBs. Although palpitations and angioedema are listed for felodipine, their signal strength is much higher compared to the other DHP-CCBs. Conclusions: The ADR risk profiles of the four DHP-CCBs differ significantly. This study identified several high-risk adverse events not included in current labels. Clinical use should consider each drug’s risk profile and patient-specific factors, with particular attention to serious risk signals. For pregnant and postpartum women, the benefits and risks of using nicardipine and nifedipine should be carefully evaluated. Full article

Background and Objectives: Despite critically low levels of chronic kidney disease (CKD) awareness worldwide, there is no psychometrically validated instrument to comprehensively assess CKD awareness across socioecological levels. This study aimed to develop, psychometrically evaluate and validate a multidimensional awareness scale grounded in socioecological theory for patients with stage 3–4 CKD. Materials and Methods: This methodological study enrolled 908 stage 3–4 CKD patients. Scale development proceeded through systematic stages: comprehensive literature review, qualitative interviews (n = 15), expert panel evaluation (n = 25), and pilot testing. The initial 72-item pool was refined to 41 items (Content Validity Index = 0.912). The sample was randomly split for exploratory factor analysis (EFA; n = 454) and confirmatory factor analysis (CFA; n = 454). Psychometric evaluation encompassed internal consistency (Cronbach’s α, McDonald’s ω), test–retest reliability (n = 30; 4-week interval), convergent validity (average variance extracted [AVE], composite reliability [CR]), discriminant validity (Fornell–Larcker criterion), and criterion validity (correlation with Turkish Health Literacy Scale-32 [TSOY-32]). Results: EFA revealed a seven-factor structure with an acceptable explained variance of 43.8%. Following iterative item elimination based on communalities (h² < 0.20) and factor loadings (λ < 0.30), CFA confirmed the final 34-item model with good fit (CFI = 0.972; RMSEA = 0.070 [90% CI: 0.067–0.074]). The factor structure captured awareness across core socioecological levels (Individual, Interpersonal/Institutional, Community, and Systemic), complemented by Treatment Adherence and Social Impact dimensions. Internal consistency coefficients were α = 0.884 and ω = 0.889 for the total scale. Test–retest reliability yielded an ICC of 0.954 (95% CI: 0.907–0.978). Convergent and discriminant validity were confirmed via composite reliability (CR: 0.740–0.953) and the Fornell–Larcker criterion. Criterion validity analysis revealed a significant correlation with TSOY-32 (r = 0.810, p < 0.001). Conclusions: The CKD Awareness Scale (CKD-AS-34) represents a novel, psychometrically validated, multidimensional awareness instrument for CKD. This scale enables clinicians to identify awareness deficits spanning individual to systemic levels, facilitating personalised patient education and targeted public health interventions. Full article

Background/Objectives: Retinoblastoma represents the most common intraocular malignancy in childhood; however, the clinical applicability of mitomycin C (MMC) is restricted by dose-dependent ocular toxicity. Consequently, the development of pharmacological strategies that sensitize tumor cells to MMC while allowing dose reduction remains an unmet therapeutic objective. In this context, quercetin, a bioactive flavonoid with pleiotropic anticancer properties, has emerged as a potential chemosensitizing agent. Methods: Human retinoblastoma cell lines Y79 and WERI-Rb1 were exposed to MMC and quercetin, administered either individually or in fixed-ratio combinations. Cytotoxic responses were quantified through dose–response modeling and IC₅₀ determination following 24 and 48 h of treatment. Drug–drug interactions were quantitatively characterized using the Chou–Talalay combination index (CI) approach and isobologram analysis. Cell cycle distribution was assessed by propidium iodide (PI)-based flow cytometric analysis to evaluate treatment-associated alterations in cell cycle progression. Apoptotic cell death was assessed by Annexin V-FITC/PI flow cytometry, while transcriptional modulation of genes associated with apoptosis, cell cycle regulation, and oxidative stress (BAX, BCL-2, TP53, CASP3, CDKN1A, and HMOX1) was evaluated by qRT-PCR. Modulation of tumor-supportive signaling was examined by measuring VEGF and IL-6 secretion. Translational relevance was further investigated using a three-dimensional (3D) tumor spheroid model, and the functional contribution of reactive oxygen species (ROS) was interrogated through N-acetyl-L-cysteine (NAC) rescue experiments. Results: Quercetin significantly enhanced the cytotoxic activity of MMC in both retinoblastoma cell lines, with CI values below 1 across IC₅₀–IC₉₀ effect levels, indicating a synergistic pharmacological interaction. PI–FACS analysis revealed that combined MMC and quercetin treatment induced a pronounced accumulation of cells in the G2/M phase, consistent with cell cycle arrest, with a more marked effect observed in Y79 cells compared with WERI-Rb1 cells. Combination treatment resulted in a pronounced increase in apoptotic cell populations compared with single-agent exposure and triggered a coordinated pro-apoptotic transcriptional response, characterized by increased expression of BAX, TP53, CASP3, CDKN1A, and HMOX1, alongside suppression of BCL-2 and a marked shift in the BAX/BCL-2 ratio. Concurrently, VEGF and IL-6 secretion were significantly reduced, reflecting attenuation of pro-angiogenic and pro-inflammatory signaling. Notably, synergistic cytotoxicity was maintained in 3D tumor spheroids, where combined treatment induced spheroid shrinkage, architectural disruption, and reduced viability. NAC pretreatment diminished ROS accumulation and partially restored cell viability, indicating that oxidative stress contributes to, but does not solely account for, the observed synergistic cytotoxic effect. Conclusions: Collectively, these findings indicate that quercetin appears to function as an effective chemosensitizing adjuvant to MMC in retinoblastoma models, through transcriptional changes consistent with p53-associated apoptotic signaling at the transcriptional level, G2/M cell cycle arrest, and partial involvement of ROS-related cellular stress responses, along with suppression of tumor-supportive signaling pathways. The preservation of synergistic activity in 3D tumor spheroids supports the potential preclinical relevance of this combination. However, these findings are based on transcriptional and phenotypic analyses and should be interpreted as hypothesis-generating, requiring further validation through protein-level and in vivo studies before translational application. Full article

The objective of the present study is to evaluate the effects of lactation stage, age, somatic cell count (SCC), and daily milk yield on plasmin–plasminogen (PL–PG) system activity and physicochemical milk traits in intensively reared Chios and Frizarta ewes. A total of 52 purebred ewes (26 ewes per breed and farm) were randomly selected and prospectively monitored during the 3rd, 5th, and 6th month post-lambing. Daily milk yield and body condition score (BCS) were recorded, and individual milk samples were collected for the assessment of PL–PG activities using enzymatic assays, SCC, electrical conductivity (EC), refractive index (RI), and pH. Correlation analysis and mixed linear regression models were used for the assessment of the effects. Lactation stage significantly affected PL–PG system traits in both breeds, but in opposite direction; plasmin and plasminogen plus plasmin declined toward late lactation in Chios ewes, whereas it increased in Frizarta ewes. Lower SCC was associated with reduced plasmin system activity in Chios ewes, whereas no effect was observed in Frizarta ewes. The plasminogen-to-plasmin ratio remained stable across lactation, breeds, and SCC classes, indicating coordinated regulation of the system. BCS was positively associated with plasmin activity during late lactation, suggesting a stage-dependent metabolic modulation. EC and pH were closely associated with SCC, while RI mainly reflected compositional variation. Our findings underline that, although the PL–PG system is primarily affected by lactation stage and mammary health status in sheep, there are breed-specific regulatory patterns which should be further investigated. Full article

Historical small towns (HSTs) embody irreplaceable cultural heritage and territorial identity, facing depopulation, economic marginalization, and infrastructure decay. Improving their liveability and attractiveness is essential to reverse these trends and boost sustainable development. In this context, HSTs are potential drivers of circular and sustainable socio-technical systems, where the circular economy (CE) offers a framework for local sustainability. However, HSTs lack adequate sustainable CE implementation tools. This study, the culmination of the H-SMA-CE project, develops a Decision Support System (DSS) to assist local policymakers in planning CE transitions in Italian HSTs. The DSS integrates three building blocks: context analysis (metabolic flows, stakeholder networks), an intervention library with cost–benefit data, and a composite Municipal Circular Economy Index (MCEI). The tool enables users to assess baseline circularity, simulate scenarios, and identify optimal investment portfolios through multi-objective optimization. This approach allows for the simultaneous evaluation of the benefits of each sustainability aspect, i.e., environmental, economic and social. Tested on the municipality of Taurasi (Italy), an HST with a wine-based economy, the results show that balanced intervention strategies yield greater circularity improvements than single-objective approaches. The paper contributes to the discourse on digital tools for sustainability transitions, offering a replicable model for evidence-based CE governance in heritage-rich territorial contexts. Full article