Search Results (16,403)

Mycotoxins are the most frequently occurring natural contaminant in food and feed products. Through the deployment of diverse agricultural strategies or biological, chemical, or physical treatments of crop products, mycotoxin contamination remains a persistent issue for the agricultural sector and food/feed industry. We previously suggested that halophytes, thanks to their high antioxidant activity, could protect animal cell lines from mycotoxin contamination. Here, a hydroalcoholic extract of Calystegia soldanella L. leaves was evaluated for in vitro total antioxidant capacity (TAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-quenching bioassays, as well as anti-inflammatory (ELISA measurement of IL-8 secretion), ROS-inhibiting production (CellROX Green assay), and calcium influx restoration (fluorescent probe Fura2-QBT assay) activities in two animal cells upon mycotoxin intoxication. C. soldanella extract displayed high antioxidant activities (DPPH IC₅₀ < 80 μg·mL⁻¹ and TAC of 90 mg AAE·g⁻¹ DW. Moreover, it exhibited a significant protective action on renal (MDBK) and intestinal (IPEC-J2) cells against zearalenone (ZEA) or T2-toxin contamination, restoring about 75% of cell viability (MTS bioassay) at 1 μg·mL⁻¹. This effect was accompanied by strong anti-inflammatory, ROS-inhibition, and membrane integrity restoration activities. A bio-guided study revealed that the fraction of C. soldanella extract eluted from C18-bound silica with 60% methanol was the most active one. Upon HPLC and 1D- and 2D-NMR analyses, major compounds identified in this fraction were flavonol-type flavonoids, including quercetin-3-O-glucose (X1), quercetin-3-O-rutinoside (X2), and quercetin-3-O-glucose-6″-acetate (X3). Enriched sub-fractions containing these compounds largely contributed to the cytoprotective effects of C. soldanella, supporting its potential use as a food/feed ingredient. Full article

Background/Objectives: Tick-borne encephalitis (TBE) is the most important tick-borne viral central nervous system (CNS) infection in Europe and Asia. Since the introduction of a vaccine in Austria in the late 1970s, sero-epidemiological studies on the true incidence of tick-borne encephalitis virus (TBEV) infection in the population have been difficult, because it was not possible to distinguish between vaccine- and infection-induced antibodies. The goal of our study has been to analyze the sero-epidemiology of TBEV infections, vaccination protection rate, and manifestation index of the disease in the region of interest. Methods: Applying a newly developed anti-TBEV-NS1-IgG assay and the neutralization test, the protection and infection rates in blood donors of the Austrian Federal State of Upper Austria.It is one of the first areas in Austria, where the TBEV vaccine had been rolled out and broadly used. Samples from blood donors of all districts of the Federal State of Upper Austria were screened for anti-TBEV-IgG. Positive sera were differentiated for infection- and vaccine-induced antibodies. The results were matched with donor age, gender, and geographical origin. Results: 2162 samples were analyzed. A total of 87.0% of the blood donors tested showed anti-TBEV-IgG related to past TBEV vaccination. Within the unvaccinated group, a total of 13.3% of male and 9.9% of female blood donors exhibited anti-TBEV-NS1-IgG, indicating a past TBEV infection. The anti-TBE-NS1-IgG seroprevalence was determined at 74/100,000 for the whole population and at 594/100,000 in the non-vaccinated population. The manifestation index (MI) was calculated at 2.8%. The MI is defined as the probability or percentage of infected individuals who develop clinical symptoms of a disease. Conclusions: Our data provide evidence of a continuing high risk of TBEV infection in the Federal state of Upper Austria. The non-vaccinated population has an eightfold higher infection prevalence compared to the whole population. The MI of TBEV for severe infection seems lower as detailed in previous reports. Full article

NKAP (NF-kappa-B-activating protein) is a ubiquitously expressed nuclear protein involved in multiple biological processes. Males with missense NKAP mutations have been reported to present with marfanoid features and behavioral and musculoskeletal abnormalities. We have previously reported that a disruptive NKAP mutation resulted in extremely skewed X chromosome inactivation (XCI), leading to phenotypic manifestation of hemophilia A (HA) in a HA carrier. In this study, with the aim of exploring the phenotypic manifestations of deleterious NKAP mutations in males, as well as their involvement in the mechanism of XCI regulation in females, we generated NKAP mutant mice using CRISPR/Cas9 technology. Gait analysis studies conducted in male mice hemizygous for mutant NKAP by the CatWalk XT system revealed significant alterations in gait parameters, consistent with hypotonia reported in human mutant NKAP patients. By breeding mutant NKAP mice with HA mice, we generated a double heterozygous mutant NKAP/HA mouse model, i.e., female mice carrying mutant NKAP with a WT F8 copy on one X chromosome, and WT NKAP with a mutant F8 copy on the other X chromosome. XCI pattern analysis using methylation-sensitive restriction enzymes demonstrated that mutant NKAP/HA females exhibited significant XCI skewing of the X chromosome bearing the mutant NKAP copy. Furthermore, these females exhibited significantly reduced F8 mRNA levels and FVIII (factor VIII) antigen levels, as demonstrated by quantitative RT-PCR and ELISA, respectively. Murine embryonic fibroblasts (MEFs) derived from a hemizygous mutant NKAP embryo exhibited markedly reduced proliferation rate and increased senescence compared to WT NKAP MEFs, suggesting that XCI skewing induced by mutant NKAP results from secondary selection against cells with an active X chromosome bearing the mutant NKAP copy. Full article

Background: Rift Valley fever virus (RVFV) is a significant mosquito-borne zoonotic virus with high public health and veterinary importance in Africa and the Middle East. Reliable diagnostic assays for detecting antibodies and assessing their functional neutralizing capacity are essential for surveillance programs, vaccine monitoring, and outbreak preparedness. Objective: This study evaluates and compares the analytical performance of a competitive enzyme-linked immunosorbent assay (cELISA) and a virus neutralization test (VNT) for detecting RVFV antibodies in vaccinated sheep sera, establishing an integrated laboratory workflow for virus titration, serological detection, and functional neutralization. Methods: Twenty serum samples were collected from sheep pre-vaccination and one month post-vaccination with Smithburn live attenuated RVFV vaccine. Sera were tested using a commercial multispecies RVFV competitive ELISA to detect antibodies specific to the viral nucleocapsid protein. Viral titration was conducted in Vero cells, and 50% tissue culture infective dose (TCID₅₀/0.1 mL) was calculated using the Reed and Muench method. VNT was performed at 24, 48, 72, and 96 h after infection with different viral doses (10² to 10⁵ TCID₅₀/0.1 mL), and the neutralizing ability of serial serum dilutions (1:2 to 1:1024) was tested. Compared with the control, protection was determined by cytopathic effect (CPE) inhibition. Results: ELISA revealed robust antibody signals up to a 1:32 dilution, with signal-to-noise (S/N) < 40%, whereas for higher dilutions, antibody detection became inconclusive or negative. Virus titration was performed to verify a stock concentration of 10^6.5 TCID₅₀/0.1 mL. The VNT exhibited time- and dose-dependent kinetics; high protection rates (≥97) were observed at 1:2–1:8 dilutions against 10²–10³ TCID₅₀/0.1 mL challenge doses; however, neutralizing efficacy decreased significantly at higher viral loads and higher serum dilutions. While cELISA and VNT results correlated strongly at low serum dilutions, the cELISA showed decreased sensitivity at dilutions ≥ 1:64, where the VNT remained capable of detecting functional neutralizing activity. Conclusions/Discussion: The results demonstrate that while both assays correlate well at high antibody concentrations, they diverge at lower concentrations. This discrepancy highlights the functional difference between binding antibodies (N-protein) and neutralizing antibodies (Gn/Gc glycoproteins). Consequently, the cELISA is ideal for rapid screening, whereas the VNT is indispensable for confirming functional immunity. Integrating both assays provides a more accurate immunological profile for RVFV surveillance and vaccine evaluation. Full article

Porcine epidemic diarrhea (PED), caused by the porcine epidemic diarrhea virus (PEDV), is an acute and highly contagious intestinal disease that inflicts substantial economic losses on the global swine industry. The nucleocapsid (N) protein of PEDV plays a critical role during viral infection and replication. In this study, the full-length N gene was cloned and expressed using the prokaryotic expression vector pET-32a (+). The purified recombinant N protein was used to immunize BALB/c mice. Subsequently, splenocytes from the immunized mice were fused with SP2/0 cells, and hybridoma cell lines secreting monoclonal antibodies (mAbs) against N protein were screened via indirect ELISA. The linear B-cell epitopes recognized by the mAbs were mapped using truncated N protein fragments. Results showed that three stable hybridoma cell lines (1A3, 1G1 and 1A10) secreting N protein-specific mAbs were obtained. Epitope mapping revealed that mAbs 1A3 and 1G1 recognized the epitope ⁷¹SNWHF⁷⁵, whereas mAb 1A10 recognized ⁶⁶RIEQP⁷⁰. Bioinformatics analysis indicated that these epitopes are highly conserved among the analyzed PEDV strains and show no cross-reactivity with the N proteins of other coronaviruses. These findings could provide valuable experimental materials for further investigation of the N protein’s structure and function and support the development of diagnostic assays and subunit antigen vaccine for PEDV. Full article

Background/Objectives: Bone metastases are a common complication of breast cancer. In our previous study, we reported that extracellular vesicles released by osteotropic human (MDA-MB-231) and murine (4T1) breast cancer cells disrupt bone homeostasis by enhancing osteoclast differentiation and impairing osteoblast function. Based on these findings, we investigated whether microRNAs contained within tumour-derived EVs could mediate these bone-altering effects. Methods: MDA-MB-231- and 4T1-EVs were tagged with the RNA-specific fluorophore SYTORNA and employed to treat mouse primary bone marrow macrophages (BMMs) and osteoblasts (OBs). We also performed RNAseq on MDA-MB-231- and 4T1-EVs to assess their miRNAs content. Finally, we evaluated the effect of selected miRNA-mimics on OBs, BMMs and HUVEC cells. Results: Fluorescence microscopy demonstrated EV-RNAs shuttling to recipient cells, while RNA sequencing on MDA-MB-231- and 4T1-EVs revealed that, of the top 20 expressed miRNAs, 10 were common. Among them, we first focused on the following four: miR-26a-5p, miR-24-3p, miR-29a-3p, and miR-29b-3p, which were linked to bone biology. We confirmed their presence in MDA-MB-231-/4T1-EVs by qPCR. Then, we evaluated their EV-mediated shuttling to BMMs and OBs using affinity tags. Among all the conditions tested, miR-29a and miR-29b were the best-shuttled miRNAs, with efficiency between 50–100% in both OBs and BMMs, both for MDA-MB-231- and 4T1-EVs. Finally, to test whether miR-29a and miR-29b could have a functional role in bone cells, OBs were transfected with miR-29a and 29b-mimics, discovering that this treatment reduced collagen1α1 and 1α2 mRNA as well as the OBs’ mineralisation ability, while the same miRNA mimics were found to have no effect on osteoclastogenesis or on in vitro angiogenesis. Conclusions: MDA-MB-231- and 4T1-EVs shuttle miRNAs to bone cells, which likely contributes to OBs’ activity impairment. Full article

Background: Cutaneous melanoma metastases (CMM) represent a clinically relevant manifestation of advanced melanoma and may constitute the first sign of disseminated disease. Their diagnosis is challenging because CMM shows highly variable clinical and dermoscopic presentations and frequently mimic other benign or malignant skin lesions. Although dermoscopy is routinely used to improve skin lesion assessment, dermoscopic criteria specific to CMM remain poorly defined and still non-standardized. Methods: We performed a narrative review of the literature to summarize dermoscopic features reported in CMM. MedLine (via PubMed) and Web of Science were searched up to 3 December 2025 using the keywords “dermoscopy” and “melanoma metastasis,” complemented by manual reference screening. Eligible studies were English-language full-text articles in peer-reviewed journals providing a complete dermoscopic description. Extracted data included patient demographics and major dermoscopic criteria, categorized as global patterns and focal dermoscopic and vascular structures. Due to heterogeneity, results were synthesized descriptively. Results: Twenty studies were included, comprising 774 patients. Dermoscopic findings were markedly heterogeneous. Globally, lesions frequently showed homogeneous pigmentation with variable colors and included amelanotic presentations. Commonly evaluated focal features included irregular dots and globules, crystalline structures, peripheral gray dots, and lacuna-like areas. Vascular patterns were prominent, particularly serpentine and corkscrew-like vessels. Conclusions: CMM dermoscopy is characterized by substantial heterogeneity and a lack of standardized criteria. Systematic classification of recurring dermoscopic features may improve diagnostic consistency and provide an interpretable framework for future artificial intelligence-based approaches supporting non-invasive recognition of melanoma metastases. Full article

This study aimed to establish an indirect ELISA for detecting the avian reovirus (ARV) epidemic strain xj-1.1 by using the purified recombinant protein pET-σC as the coating antigen. To optimize assay performance, key parameters were systematically evaluated, including antigen-coating concentration, serum dilution, blocking reagent and duration, serum incubation time, and the dilution and reaction time of the HRP-conjugated secondary antibody. The optimized conditions identified were a coating antigen dilution of 1:100, serum dilution of 1:1600, coating at 37 °C for 1 h followed by overnight incubation at 4 °C, and blocking with 5% skim milk for 2 h. The optimal serum incubation time was 1.5 h, with the secondary antibody diluted 1:1000 and incubated for 2 h, followed by a 20-min color development step. The cut-off value for distinguishing positive and negative samples was determined to be 0.121. Validation of the assay demonstrated favorable specificity, sensitivity, and repeatability, indicating that the developed indirect ELISA provides a reliable method for detecting ARV xj-1.1 infection. Full article

Cognitive decline represents one of the most common clinical manifestations of neurodegenerative diseases (NDs), substantially affecting the quality of life of both patients and their families. Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis are major NDs characterized by a progressive degeneration of the central nervous system, with functional impairments extending beyond motor symptoms to multiple cognitive domains, including memory, attention, language, and executive functions. Increasing evidence highlights misfolded protein accumulation as a key driver of neuronal dysfunction and cognitive deterioration. This narrative review examines the major cognitive deficits associated with these disorders, focusing on the underlying molecular mechanisms, particularly protein aggregation, as well as clinical manifestations and their effects on daily life. Furthermore, current diagnostic tools and emerging therapeutic options for mitigating cognitive decline will be further discussed. Full article

Periprocedural myocardial infarction after percutaneous coronary intervention (PCI) remains a debated entity, especially in the era of high-sensitivity cardiac troponin assays, which frequently detect biomarker rises even when clinically meaningful ischemia is absent. This review critically examines the main contemporary frameworks used to define these events, including the Fourth Universal Definition of Myocardial Infarction (UDMI), the Academic Research Consortium (ARC)-2 consensus, and the Society for Cardiovascular Angiography and Interventions (SCAI) definition, comparing biomarker thresholds, requirements for objective evidence of ischemia, and procedural criteria. We discuss how differences among definitions shape reported event rates and contribute to heterogeneity in event adjudication across studies. Key pathophysiologic mechanisms of myocardial injury during PCI are summarized, including side-branch compromise, distal embolization, microvascular dysfunction, and mechanical complications. Particular attention is given to the limitations of current criteria, such as incomplete assay standardization, variability in sampling timing, inconsistent reliability of ancillary criteria, including electrocardiography and imaging, and an uneven relationship between biomarker elevation and subsequent outcomes. Finally, we outline priorities for future updates, including harmonization of biomarker thresholds, greater emphasis on relative biomarker dynamics, and structured adjudication that integrates biomarkers with objective ischemic evidence. These steps may improve diagnostic specificity, reduce misclassification, and strengthen the clinical and trial relevance of periprocedural ischemic endpoints. Full article

Leadership plays a central role in the long-term sustainability of family enterprises, yet existing evidence is fragmented across contexts and methodologies. This systematic review synthesizes empirical findings on leadership practices that support sustainable family entrepreneurship. The objectives are to identify available evidence on leadership in sustainable family enterprises, describe the methodologies employed, and examine how leadership is perceived and enacted across global contexts. The review followed PRISMA 2020 guidelines. Searches were conducted on 28 January 2025 on Scopus, Web of Science, EBSCOHost, and ProQuest. Eligibility criteria included empirical studies, full accessibility, publication in English, non-duplicated records, and relevance to leadership in sustainable family enterprises. Twenty-six studies met the inclusion criteria. Data extraction focused on context, methodology, leadership evidence, and key findings. Studies spanning Asia, Europe, Latin America, Africa, and the Middle East indicate that leadership strongly shapes sustainability outcomes in family firms. Three core leadership dimensions emerged: transformational leadership, which promotes innovation, engagement, affective commitment, and continuity; transactional leadership, which supports governance, succession planning, operational control, and performance; and ethical leadership, which fosters trust, shared values, and social responsibility. Cross-cutting themes include gendered leadership contributions, succession risk management, and cultural influences. Sustainable family enterprises rely on multidimensional leadership integrating these approaches, reinforced by structured succession processes, value alignment, and human capital investment. Full article

Carbapenemase-producing bacteria, particularly those expressing the KPC-3 variant, pose a critical global health threat due to their resistance to nearly all β-lactam antibiotics, including carbapenems. Rapid and reliable detection tools are urgently needed to improve infection control and guide patient management. Nanobodies (VHHs) present a promising alternative to conventional antibodies thanks to their high stability, small size, and capacity to access cryptic epitopes. Here, we report the generation and characterization of a nanobody specifically targeting KPC-3. An immune VHH phage display library was constructed, with over 90% of clones containing correctly sized inserts. After three rounds of biopanning, high-specificity binders were identified by ELISA screening. Sequencing identified a nanobody with hallmark VHH features, which was expressed and validated by ELISA and Western blot. Although kinetic assays showed no inhibition of KPC-3 enzymatic activity, interestingly, the nanobody demonstrated high-binding recognition of both KPC-2 and KPC-3 in periplasmic extracts from clinical strains. Structural modeling further supported these results, highlighting favorable interaction surfaces. This study provides the first evidence of a nanobody raised against KPC-3 that recognizes a conserved epitope shared by KPC-3 and KPC-2, underscoring its promising use as a molecular tool for detecting KPC variants and establishing a basis for future affinity maturation toward therapeutic applications. Full article

Background/Objectives: Porcine epidemic diarrhea (PED) is a highly contagious enteric disease caused by porcine epidemic diarrhea virus (PEDV) and is associated with severe clinical signs and high mortality in neonatal piglets. Vaccination is an important strategy for PED control through the induction of humoral immunity. This study aimed to compare immune responses induced by inactivated and live-attenuated PEDV vaccines and to evaluate a heterologous prime-boost vaccination strategy in PEDV-naïve replacement gilts. Methods: Twenty-four PEDV-naïve replacement gilts were randomly assigned to four groups: unvaccinated control, inactivated vaccine administered twice (K/K), live-attenuated vaccine administered twice (L/L), and live-attenuated priming followed by an inactivated booster (L/K). Pigs received two intramuscular vaccinations at 16 weeks of age and two weeks later. Serum samples collected up to 42 days post-vaccination were analyzed for PEDV-specific IgG and IgA antibodies by ELISA, and serum-neutralizing antibody titers were determined using a serum neutralization test. Results: The L/K regimen induced the highest PEDV-specific IgG responses, with peak levels at day 28 post-vaccination that were significantly higher than those in the K/K and control groups. Serum-neutralizing antibody titers were significantly higher in the L/K and L/L groups than in the K/K and control groups. Serum IgA responses were low and transient across all vaccination groups. Conclusions: A heterologous prime-boost vaccination strategy using a live-attenuated PEDV vaccine followed by an inactivated booster induces strong systemic humoral immune responses in replacement gilts and represents a promising approach for PEDV vaccination programs. Full article

Background and Objectives: Pediatric and adolescent obesity is a growing global health concern that is often associated with cardiometabolic comorbidities. Lifestyle interventions represent first-line therapy; however, many adolescents with moderate-to-severe obesity fail to achieve clinically meaningful weight loss. The objective of this review is to provide a comprehensive overview of surgical and endoscopic interventions for adolescent obesity. Materials and Methods: A structured search of PubMed, Scopus, Web of Science and the Cochrane Library was conducted. Studies reporting outcomes of bariatric surgery (sleeve gastrectomy (SG), Roux-en-Y gastric bypass (RYGB), adjustable gastric banding (AGB)) and endoscopic interventions (endoscopic sleeve gastroplasty (ESG) and intragastric balloons (IGBs)) in patients ≤ 21 years were included. Data on weight loss, BMI reduction, metabolic outcomes, adverse events and follow-up were extracted and summarized. Results: Bariatric surgery remains the most effective intervention, with SG and RYGB achieving substantial and durable weight loss and high rates of comorbidity remission. ESG is an emerging intervention with preliminary short-term efficacy and safety data, though evidence is limited to small pediatric cohorts. IGBs provide reversible, non-surgical weight reduction with consistent short-term efficacy, but long-term durability is variable and largely dependent on adherence to behavioral programs. Across all interventions, psychosocial support, family involvement and multidisciplinary care significantly influence the outcomes. Conclusions: Procedural interventions play a pivotal role in adolescents with moderate-to-severe obesity. IGBs could represent a minimally invasive, reversible option, particularly as bridging or adjunctive therapy. Prospective, long-term studies are needed to optimize patient selection, evaluate developmental safety and determine sustainable outcomes. Full article

Tracking highly maneuvering, non-cooperative UAVs poses significant challenges due to rapid and unpredictable changes in target dynamics. Under such conditions, traditional single-model filters often fail to maintain reliable state estimates, resulting in degraded tracking performance. Multiple-Model Kalman Filter ( (MMKF) approaches, including the Generalized Pseudo Bayesian (GPB1) and Interacting Multiple-Model (IMM) algorithms, improve robustness by simultaneously considering multiple candidate motion models and weighting them according to the observed target behavior. Adaptive strategies, such as ${\chi }^2$-test-based or t-test-based methods, further enhance performance by dynamically responding to changes in maneuvering patterns. This paper presents a multi-criteriacomparative assessment of four MMKF formulations–GPB1, IMM, ${\chi }^2$-test-based, and t-test-based filters– under a consistent modeling and simulation framework. Particular emphasis is placed on systematically analyzing the role of the transition probability matrix (TPM), investigating how fixed, adaptive, and TPM-free strategies affect estimation accuracy, robustness to noise, and mode-identification performance. Beyond conventional Root Mean Square Error (RMSE) metrics, the filters’ comparison is carried out through confusion matrices and dwell time analysis to highlight performance nuances and trade-offs. This allows to establish which filter formulation is preferable in different operational conditions. Full article