Search Results (17,661)

Primula vulgaris Huds., one of the 33 Primula L. species native to Europe, occurs across diverse habitats, including the biodiversity hotspot of the Prespa Lake region (NW Greece). Building on previous phytochemical studies, the present work provides the first detailed characterization of flavonoids from the aerial parts of the species growing wild in the area. Using classical chromatographic separation methods combined with spectrometric techniques, seven metabolites were isolated and structurally elucidated from the dichloromethane and methanol extracts. These included flavone (1), 2′-methoxyflavone (2), 3′-methoxyflavone (3), 3′-hydroxy-4′,5′-dimethoxyflavone (4), kaempferol-3-O-β-glucopyranosyl-(1→2)-β-glucopyranosyl-(1→6)-β-glucopyranoside (6), 3′-hydroxyflavone-4′-O-β-glucopyranoside (7) and 5,6,2′,3′,6′-pentamethoxyflavone (5), which was reported for the first time in this species. Additionally, the total phenolic content (TPC) of the methanol extract was determined using the Folin–Ciocalteu method, demonstrating 46.46 ± 2.48 mg GAE/g extract, while through the DPPH radical scavenging assay, it expressed moderate activity. Overall, these results provide novel insights into the flavonoid composition of Greek P. vulgaris and support its potential for further pharmacological investigations and herbal applications. Full article

Background: This study aimed to assess the presence of thermotolerant Campylobacter resistant to ciprofloxacin and erythromycin in poultry slaughterhouses and retail markets, as well as to characterize their multidrug resistance profiles, genetic determinants, and clonal relationships. Methods: Samples were collected at slaughterhouses from cecal content (n = 270), neck skin (n = 270), and wastewater (n = 9), and at retail markets from breast skin (n = 241). Isolates were obtained from mCCDA agar supplemented with ciprofloxacin (2 μg/mL) and identified as C. jejuni or C. coli by PCR. The agar microdilution test was used to determine the minimum inhibitory concentration for ciprofloxacin and erythromycin, and other critical antibiotics. Point mutations in gyrA (Thr86Ile) and 23S rRNA (A2075G), virulence genes (flaA, flhA, cadF, and cdt), and clonal relationships were assessed by PCR and PFGE. Results: At the slaughterhouses, thermotolerant Campylobacter spp. resistant to erythromycin and ciprofloxacin were detected in 48.55% (107/549) of the samples, whereas 4.56% (11/241) of retail samples were positive. The Thr86Ile substitution in gyrA and the A2075G mutation in the 23S rRNA gene were detected in 92.97% and 89.84% of the isolates, respectively. Most isolates (>80%) were multidrug resistant and harbored key virulence genes (flaA, flhA, and cadF). C. jejuni exhibited the highest prevalence of cdt genes (76.19%). There was substantial genotypic diversity among isolates, with broad distribution across the sampled matrices and sites. Conclusions: These findings highlight the circulation of multidrug-resistant and potentially virulent thermotolerant Campylobacter spp. in the later stages of the poultry meat supply chain. Full article

Background/Objectives: Galleria mellonella (G. mellonella) larvae have emerged as a valuable in vivo model for antifungal drug screening. This study aimed to determine the optimal inoculum concentrations of Candida albicans (C. albicans) in G. mellonella, as well as the appropriate fluconazole concentrations, in order to standardize a preliminary screening method for compounds with antifungal activity. Methods: Larvae were infected with four C. albicans strains, including two reference strains (ATCC^® 10231 and ATCC^® 90028) and two oral isolates (A1 and A2). Fluconazole toxicity was evaluated at doses of 20, 40, and 80 mg/kg over a 72 h period. In the treatment assays, larvae were infected via the left pro-leg and treated with fluconazole, administered as a single or two doses, one hour after infection. Larval viability was monitored over five days based on movement, cocoon formation, and melanization, and survival data were analyzed using Kaplan–Meier curves and the log-rank test. Results: Fluconazole showed no toxicity at the tested concentrations. Infection with up to 2 × 10⁷ cells/mL was non-lethal for most strains, except for A2, which exhibited 50% mortality within 48 h but it was effectively controlled with a single 20 mg/kg dose of fluconazole. Infection with 2 × 10⁸ cells/mL resulted in complete mortality within 48 h; however, a single 80 mg/kg dose significantly improved survival. Conclusions: The G. mellonella model proved to be a suitable and reproducible in vivo system for the preliminary screening of antifungal compounds. The standardized experimental conditions established in this study support its applicability for evaluating antifungal activity in early research stages. Future studies should expand this approach to different fungal species and antifungal agents, as well as explore its applicability in combination therapies. Full article

Background: Bovine mastitis (BM) remains an economically significant disease in the global dairy industry. Multidrug resistance (MDR) in Klebsiella pneumoniae and Klebsiella grimontii has increased in recent years, representing an area of concern related to BM. Methods: Bovine mastitis 1-DH1 and 2-DH2 isolates (n = 2) were investigated. Antimicrobial susceptibility testing was performed using the Neg-Urine-Combo98 panel. Antimicrobial resistance genes (ARGs), virulence factor (VF) genes and plasmid replicons were identified by whole-genome sequencing (WGS). Phylogenomic analyses were performed for a visual comparison of the genomes. Results: Phenotypically, isolates 1-DH1 and 2-DH2 were identified as K. pneumoniae-1DH1 and Klebsiella oxytoca, respectively; the latter was subsequently confirmed as K. grimontii-2DH2 by WGS. K. pneumoniae-1DH1 (20.0%, 5/25) exhibited phenotypic resistance to amoxicillin–clavulanic acid, ampicillin (AM), cefuroxime, colistin (Cl), and nitrofurantoin (Fd), whereas K. grimontii-2DH2 (8.0%, 2/25) was resistant to AM and nalidixic acid (NA). In total, 31 and 32 ARGs and 10 and 15 VF genes were identified in K. pneumoniae-1DH1 and K. grimontii-2DH2, respectively. Two plasmid replicons were detected in K. pneumoniae-1DH1 (one harboring the blaCTX-M ARG) compared to one in K. grimontii-2DH2 (encoding both the blaCTX-M ARG and the astA VF gene) isolates, respectively. Phylogenomic analysis showed that K. pneumoniae-1DH1 clustered most closely with human-derived K. pneumoniae isolates, whereas K. grimontii-2DH2 grouped with environmental K. grimontii isolates. Conclusions: Novel phenotypic resistance profiles were observed in the BM-associated K. pneumoniae-1DH1 (MDR, Cl and Fd) and K. grimontii-2DH2 (AM and NA) isolates. Notably, K. grimontii-2DH2 harbored a plasmid replicon carrying both the blaCTX-M ARG and the astA VF gene. Full article

Background/Objectives: Although cyberbullying among adolescents has been widely studied, relatively little attention has been paid to the overlapping roles through which cyberbullying is experienced. This study reconceptualizes cyberbullying involvement by classifying perpetration, victimization, and witnessing into eight mutually exclusive involvement types, enabling systematic and non-overlapping comparison of adolescents’ experiences. The study further examines how engagement in individual-level cyber-risk indicators is associated with different patterns of cyberbullying involvement. Methods: The study analyzed nationally representative data from the 2022 Cyberbullying Survey conducted by the Korea National Information Society Agency, including 9693 students from elementary, middle, and high schools across South Korea. Individual-level cyber-risk indicators were assessed through multiple dimensions, including risky online behaviors, intensity of digital activity, peer environments, and awareness of harmful online behaviors. Multinomial logistic regression analyses were conducted to examine associations between individual-level cyber-risk indicators and the eight types of cyberbullying involvement. Results: Engagement in individual-level cyber-risk indicators was associated with increased odds of involvement in at least one cyberbullying type. Risky online behaviors and exposure to peers engaging in cyberbullying were linked to higher likelihood of both single and overlapping involvement patterns, whereas greater acceptance of harmful online behaviors was consistently associated with lower odds of victimization. Conclusions: These findings underscore cyberbullying as a relational and context-dependent phenomenon shaped by everyday digital practices and peer norms rather than isolated individual behavior. From a school social work perspective, the results support preventive, environment-focused interventions, including school-based media literacy education and institutionalized cyberbullying response systems, as promising strategies for reducing cyberbullying involvement among adolescents. Full article

While Microservice Architectures (MSAs) offer enhanced scalability and maintenance, they introduce significant complexity for access control and, specifically, the rigorous enforcement of the Principle of Least Privilege (PoLP). This lack of clear privilege boundaries is a major security vulnerability in microservice-based systems. To address this gap, this study conducts a systematic mapping study to provide a comprehensive guide and taxonomy on implementing PoLP in MSA. We identify and categorize existing mechanisms, best practices, and the technical and non-technical challenges encountered during implementation. The systematic search identified 25 primary studies, revealing a significant contribution from journal venues, particularly Computers & Security. Key findings detail the top technical challenges, including performance overhead, fragile container isolation, and authentication/authorization gaps inherent in service-to-service communication. Proposed mechanisms are categorized into four groups: policy and access control, code and configuration hardening, runtime/kernel-level methods, and general frameworks. Similarly, organizational challenges are grouped by people/culture, tooling/architecture, process/governance, and resource/expertise. This study provides a valuable roadmap and taxonomy for diverse security stakeholders. The identified research gaps—concerning AI integration, DevSecOps adoption, education, and dynamic analysis—underscore the need to shift from the currently predominantly theoretical approaches towards practical, experimental research to advance the real-world application of PoLP. Full article

Background/Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) continues to pose a significant challenge for infection prevention, particularly because of its ability to persist on surfaces and form resilient biofilms. Although bacteriophages have attracted renewed interest as alternatives or complements to chemical disinfectants, their applied use requires careful assessment of antimicrobial performance, formulation tolerance, and genomic context. Methods: Staphylococcus-infecting bacteriophages were isolated from environmental sources and examined against reference Staphylococcus isolates. Two phage isolates, designated MRSA-W3 and SA-W2, displayed lytic activity against a broad subset of clinical MRSA strains. Using a time-resolved agar-based infection assay, phage exposure resulted in a multiplicity-of-infection-dependent decline in viable MRSA populations. Results: Time-resolved infection assays revealed a multiplicity-of-infection-dependent reduction in viable MRSA, with a pronounced decrease observed approximately 40 min post-infection. At this time point, phage-treated cultures showed a reduction of 1.2–1.8 log₁₀ CFU/mL relative to untreated controls (mean Δlog₁₀ = 1.5; 95% CI, 1.1–1.9), while control cultures remained stable. Quantitative biofilm assays demonstrated that both phages reduced biofilm biomass compared with untreated conditions, with inhibition values ranging from 20% to 45% across isolates (p ≤ 0.05), reflecting strain-dependent but reproducible effects. Assessment of formulation compatibility indicated that both phages retained infectivity following exposure to sodium dodecyl sulfate, Triton X-100, and Tween 80, whereas ethanol (≥10%) and higher concentrations of dimethyl sulfoxide were associated with rapid loss of activity. In surface disinfection models, selected phage–surfactant formulations achieved a maximum reduction of 2.18 log₁₀ CFU/cm² compared with untreated controls (p ≤ 0.05). Infection-coupled whole-genome sequencing of MRSA-infecting phage MRSA-W3 produced a high-quality assembly (99.99% completeness; 0.13% contamination) and revealed a mosaic genome containing incomplete prophage-like regions, which were interpreted conservatively as evidence of shared phage ancestry rather than active temperate behavior. Conclusions: Therefore, these findings suggest that bacteriophage-based approaches may be feasible for MRSA surface decontamination, while clearly emphasizing the need for context-specific validation before practical implementation. Full article

Background/Objectives: Cell-free DNA (cfDNA) is a valuable biomarker for cancer diagnosis and therapy monitoring; however, its low abundance and fragmented nature present major challenges for reliable isolation, particularly from limited plasma volumes. Here, we report the development and evaluation of a novel magnetically assisted microfluidic chip with a three-inlet design for efficient cfDNA extraction from small-volume plasma samples. Methods: The platform enables controlled infusion of plasma, lysis buffer, and magnetic nanoparticle suspensions at defined flow rates. An external magnetic field selectively captures cfDNA-bound nanoparticles while efficiently removing background impurities. Results: Direct comparison with two in vitro diagnostic (IVD)-certified commercial cfDNA extraction kits showed that the microfluidic system achieved comparable cfDNA yields at standard plasma volumes and superior performance at reduced input volumes. High DNA purity and integrity were confirmed by quantitative PCR amplification of a housekeeping gene and clinically relevant targets. The complete workflow required approximately 9 min, used minimal equipment, reduced contamination risk, and enabled rapid processing with future potential for parallel multi-chip configurations. Conclusions: These findings establish the proposed microfluidic platform as a rapid, reproducible, and scalable alternative to conventional cfDNA extraction methods. By significantly improving recovery efficiency from small plasma volumes, the system enhances the clinical feasibility of liquid biopsy applications in cancer diagnostics and precision medicine. Full article

Background: Porphyromonas gingivalis outer membrane vesicles (OMVs) are key mediators of host–pathogen interactions and have been implicated in both periodontal disease and systemic conditions, including pregnancy complications. Although OMV production and cargo are known to be influenced by environmental stress, how oxidative stress reshapes P. gingivalis OMVs and their functional impact on trophoblast cells remains poorly understood. Here, we investigated how exposure to hydrogen peroxide (H₂O₂) affects OMV biogenesis, composition, and their ability to modulate bacterial invasion in trophoblast cells. Methods: P. gingivalis was cultured anaerobically and exposed to 30 mM H₂O₂ during the final 24 h of growth. OMVs were isolated by differential ultracentrifugation and characterized by nanoparticle tracking analysis and transmission electron microscopy and OMV protein cargo was analyzed by proteomics. Functional effects were assessed using invasion and persistence assays in HTR-8/SVneo trophoblast cells pretreated with OMVs. Results: Oxidative stress did not significantly alter total OMV yield but resulted in smaller vesicles (control OMV 168.2 ± 8.7 nm vs. OMV from H₂O₂-treated cultures 130.0 ± 13.8 nm) with reduced negative surface charge and increased membrane-associated FM4-64 fluorescence. Proteomic analysis revealed a remodeling of the OMV protein cargo under oxidative stress, including the selective enrichment of a von Willebrand factor type A domain-containing protein. Functionally, OMVs from control cultures led to a 2.5-fold increase in P. gingivalis invasion and a 4-fold increase in intracellular persistence in trophoblast cells, whereas OMVs produced under oxidative stress failed to promote these processes. Conclusions: Together, these findings highlight oxidative stress as a key determinant of OMV-mediated host–pathogen interactions at the maternal–fetal interface. Full article

Rapid detection of β-lactamase activity is becoming increasingly important as β-lactam resistance spreads at an alarming rate and conventional diagnostics often require several hours to deliver actionable results. Over the past decade, methods based on luminescence, bioluminescence, chemiluminescence, and fluorescence have become powerful tools for the functional assessment of resistance resulting from β-lactamase activity. These approaches provide highly sensitive, activity-based readouts, often within minutes, and frequently rely on simple optical instrumentation. In this review, we summarize recent developments in luminescent probe design between 2015 and 2025, with emphasis on reaction mechanisms, analytical performance, and the ability of these systems to discriminate between different β-lactamases, including narrow-spectrum enzymes, AmpC, ESBL, and carbapenemases. We also discuss their applications in bacterial cultures, clinical isolates, complex biological matrices and, in some cases, in vivo models. While luminescent assays are not yet positioned to replace standard susceptibility testing, they offer a practical and increasingly robust complement to culture-based and molecular methods. The emerging trends highlighted here, such as self-immobilizing fluorogenic probes, chemiluminescent relay systems, nanomaterial-based sensors and AI-assisted mobile platforms, suggest that luminescent β-lactamase detection could play a meaningful role in future rapid diagnostics and resistance surveillance. Full article

This study evaluates how legal and regulatory architectures shape banks’ digital transformation in Qatar relative to peer jurisdictions and isolates the regulatory components that most strongly predict observed differences in digital maturity. Employing a comparative mixed-methods design, the study links a structured legal-regulatory assessment to quantitative benchmarking of fifteen banks (five Qatar, ten international) using a Digital Maturity Index and inferential tests (descriptive statistics, independent-samples t-tests, and OLS regressions). International banks exhibit higher average digital maturity than Qatar banks, and across the sample, regulatory clarity and coherence are positively and significantly associated with digital maturity, whereas supervisory intensity alone shows no comparable effect; implementation frictions in open banking/interoperability, unified data protection, and approval timelines constrain collaboration and product rollout in Qatar. Moreover, the cross-sectional design, modest sample size, and index weighting choices limit causal inference and external validity, indicating the need for longitudinal and quasi-experimental designs to corroborate mechanisms and generalize findings. Policymakers should adopt risk-proportionate, outcomes-based rules, codify interoperable API standards, strengthen data rights and cloud/third-party governance, and establish sector-level KPIs to match supervisory expectations with bank execution and accelerate safe digitalization. Enhancements to privacy, data portability, and inclusive digital onboarding are likely to improve consumer trust, competition, and access, thereby advancing broad-based participation in digital financial services. The study integrates legal analysis with bank-level operational metrics through an analytically tractable index and a Qatar–international comparison, demonstrating the outsized role of regulatory clarity in advancing digital maturity. Full article

This study investigates the physical and cyber-physical resilience of smart grids with a high share of renewable energy sources (RESs) dominated by permanent magnet synchronous generators (PMSGs). The originality of this work lies in the development and unified evaluation of five integrated control strategies, the PLL with grid following, VSG with grid shaping, VSG+BESS, VSG+STATCOM, and VSG+BESS+STATCOM, implemented within a coherent simulation framework based on Python. Unlike previous works that analyze these methods in isolation, this study provides a comprehensive quantitative comparison of their dynamic characteristics, including frequency root mean square deviation, maximum deviation, and composite resilience index (RI). To extend the analysis beyond static conditions, a multi-generator (multi-PMSG) scenario with heterogeneous inertia constants and variable load profiles is introduced. This dynamic model allows the evaluation of natural inertia diversity and the effects of inter-generator coupling compared to the synthetic inertia emulation provided by VSG-based control. The combined VSG+BESS+STATCOM configuration achieves the highest synthetic resilience, improving frequency and voltage stability by up to 15%, while the multi-PMSG system demonstrates comparable or even higher RI values due to its inherent mechanical inertia and decentralized response behavior. In addition, a cyber-physical scenario is included to evaluate the effect of communication delays and false data injection (FDI) on VSG frequency control. The results show that a communication delay of 50 ms reduces RI by approximately 0.2%, confirming that even minor cyber disturbances can affect synchronization and transient recovery. However, hybrid control architectures with local energy buffering (BESS) show superior resilience under such conditions. The main technical contribution of this work is the establishment of an integrated analytical and simulation framework that enables the joint assessment of synthetic, natural, and cyber-physical resilience in converter-dominated smart grids. This framework provides a unified basis for the analysis of dynamic stability, hybrid control interaction, and the impact of cyber uncertainty, thereby supporting the design of low-inertia, resilient, and secure next-generation power systems. Full article

Background: Cardiovascular comorbidities are major determinants of poor outcomes among patients admitted with COVID-19. However, the prognostic role of arterial hypertension alone remains uncertain. Little is known about the cumulative impact of concomitant hypertension and heart failure. This study assessed whether the combined burden of arterial hypertension and pre-existing heart failure identifies a high-risk phenotype for adverse in-hospital outcomes among COVID-19 patients. Methods: In this retrospective, real-world cohort study, 395 consecutive adults hospitalized with confirmed COVID-19 at a single infectious diseases center between March 2020 and December 2024 were included. We categorized patients into three cardiovascular phenotype groups: no hypertension or heart failure (n = 23), hypertension without heart failure (n = 193), and concomitant hypertension and heart failure (n = 178). The primary outcome was in-hospital all-cause mortality, while ICU admission served as a secondary outcome, invasive mechanical ventilation, and length of hospital stay. Multivariable logistic regression included age, sex, BMI, diabetes mellitus, and vaccination status to evaluate independent associations between the cardiovascular risk group and outcomes. Results: Overall in-hospital mortality was 7.3% (29/395). Mortality increased stepwise across the cardiovascular risk groups: 8.7% in patients without hypertension or heart failure, 3.1% in those with hypertension only, and 11.8% in patients with concomitant hypertension and heart failure (p = 0.004). In adjusted analyses, concomitant hypertension and heart failure were linked to higher adjusted odds of in-hospital death than no cardiovascular disease (odds ratio, 3.49; 95% confidence interval, 1.46–8.35). Isolated hypertension was not significantly associated with mortality. ICU admission and length of hospital stay also increased with cumulative cardiovascular burden. Patients with combined hypertension and heart failure showed more pronounced inflammatory and renal abnormalities at admission. Conclusions: Among hospitalized COVID-19 patients, the coexistence of arterial hypertension and heart failure identifies a vulnerable cardiovascular phenotype associated with higher in-hospital mortality and resource use than either no cardiovascular disease or hypertension alone. These findings support evaluating cardiovascular comorbidities cumulatively rather than in isolation. These findings are exploratory and require external validation in independent, larger multicentre cohorts. Findings may support careful use for short-term risk stratification and closer monitoring strategies during COVID-19 hospitalization. Full article

Background: Intra-abdominal infections (IAIs) are one of the leading causes of non-traumatic death in emergency surgery units. The appropriateness of empirical antibiotic therapy is fundamental for outcomes and for limiting the spread of resistance. This study aimed to assess the epidemiology and antibiotic resistance patterns of microorganisms recovered from complicated intra-abdominal infections due to colorectal perforation at an Italian University Hospital during a nine-year period. Methods: This study evaluated a cohort of patients subjected to emergency surgery for colonic perforation with collected intrabdominal fluid samples from 2015 to 2024. Patterns of isolated bacteria and antibiotic resistance status were collected and correlated to patient outcomes. Results: 321 patients were enrolled; the average age was 70.2 years. The main diagnoses were complicated diverticulitis (58%), colorectal carcinoma perforation (18%), and acute intestinal ischemia (24%). 80.4% were immunocompromised; average hospital stay (HLOS) was 15.6 days; 60.1% developed postoperative complications. Microbiological cultures were available for 111 patients: 56.7% had mono-microbial infections and 43.3% multi-microbial infections. 53 antibiotics and 9 antifungals were tested, with resistance rates exceeding 20% for many pathogens. Multivariate analyses showed that documented IAIs are associated with longer postoperative hospital stays (p 0.003 CR 8.075) but not with patient mortality (p 0.031). Prolonged HLOS was more commonly observed in patients with polymicrobial infections or infections caused by multi-drug-resistant organisms (p 0.03; p 0.003). Conclusions: Microbiological characteristics of isolated bacteria do not directly influence mortality; however, the presence of polymicrobial infections and resistant pathogens directly affects the duration of hospitalization and often leads to the development of chronic disease conditions. Full article

Background/Objectives: Antifungal resistance among filamentous fungi is an increasing global concern with significant implications for clinical management. Herein, we propose a study aiming to investigate in vitro susceptibility patterns and epidemiology of filamentous fungi in Southern Italy, focusing on MIC distributions and resistance trends. Methods: We reported susceptibility results from Aspergillus spp., Fusarium spp., and Scedosporium/Lomentospora spp. clinical isolates, which underwent azoles, echinocandins, and amphotericin B in vitro testing. Results: Aspergillus fumigatus was the most frequently isolated species, showing an alarming increase in reduced susceptibility to amphotericin B (9.1%). The highest MIC ranges for this antifungal drug emerged in the case of A. fumigatus (1–4 mg/L) and A. terreus (2–8 mg/L), while A. flavus (0.5–4 mg/L) and A. niger (0.25–4 mg/L) showed lower values. As regarding azoles, all the Aspergillus spp. strains exhibited variable MIC values, reporting a 0.06–16 mg/L MIC range for itraconazole, 0.125–1 mg/L for voriconazole, and 0.03–1 mg/L for posaconazole. Fusarium solani exhibited high MICs for azoles (8 mg/L) and amphotericin B (2–4 mg/L), while F. oxysporum and F. proliferatum showed lower MICs (0.25–2 mg/L for amphotericin B and a MIC range of 0.5–8 mg/L for posaconazole). Lomentospora prolificans and Scedosporium apiospermum demonstrated multidrug resistance across all tested antifungals, reporting MIC ranges of 4–8 mg/L for amphotericin B, 0.25–16 mg/L for posaconazole, 0.25–8 mg/L for voriconazole, and 0.125–8 for itraconazole. Conclusions: Our data highlight the critical emergence of reduced antifungal susceptibility among filamentous fungi in Southern Italy, underlining the importance of epidemiological surveillance, precise species identification, and optimized susceptibility testing in the case of mould etiology for invasive fungal infections. Full article