Search Results (6,755)

The translation of nucleic acid amplification into practical point-of-care and biosensor-integrated diagnostics is still significantly impeded by the necessity for rapid sample preparation. For this reason, a broad comparison of seven commercially available kits for DNA/RNA extraction containing their temperature-related adjustments was performed. Extracts isolated from SARS-CoV-2-positive nasopharyngeal swabs, viral stocks, as well as laboratory-prepared suspensions of clinically relevant Gram-positive and Gram-negative bacteria were evaluated by recombinase polymerase amplification (RPA) and real-time PCR. In addition, the impact of transport media for SARS-CoV-2 samples was investigated. Extraction performance varied markedly according to the kit, pathogen, sample background. For SARS-CoV-2, rapid extraction was more effective for samples collected in viral transport medium than in inactivation buffer. Across bacterial targets, performance was species dependent, highlighting substantial differences in compatibility between simplified extraction workflows and downstream amplification. Among the rapid methods tested, a simplified QuickExtract protocol (95 °C, 5 min) provided the most consistent overall results, although it did not uniformly match the reference silica-based method for all targets. In conclusion, these results demonstrate that rapid nucleic acid extraction must be thoroughly evaluated as an essential element of the entire sample-to-answer workflow, rather than being chosen as a standalone preprocessing step for point-of-care molecular diagnostics. Full article

The continuous and rapid spread of African swine fever (ASF) still poses a significant threat to Eurasian wild boar and domestic pigs, leading to substantial economic losses in all associated sectors annually. In Europe, including Germany, affected wild boar populations have become an important driver and host of ASF virus (ASFV), and monitoring and surveillance is key to tailor control measures that impede viral spread. While molecular methods are used to confirm the disease and to monitor viral evolution, serology gains importance in endemically affected regions as it provides insights into disease dynamics and possible attenuation of ASFV strains. Frontline serological diagnosis is done using ELISA assays, of which several are commercialized. However, accurate performance of ELISA assays is key for correct interpretation of wild boar samples. Due to the various stages of hemolysis and decay, field samples from wild boar can be challenging for ELISA assays. To assess which indirect or competitive ELISA kit performs best when dealing with such samples, we compared the sensitivity and specificity of four commercially available ELISA kits that are licensed in Germany, as well as three unlicensed but commercially available kits. For this comparison, we used all wild boar samples submitted to the National Reference Laboratory (NRL) for ASF in years 2021 and 2022, as well as samples from domestic pigs to have a control cohort where sample quality is optimal. We observed that wild boar samples, varying in stage of hemolysis and decay, were challenging for all kits included in this study. However, samples of domestic pigs were largely interpreted correctly by ELISA, using immunoperoxidase test as verification method. Additionally, the comparability of results obtained by regional laboratories was high. Our study provides data that highlights the importance of careful kit selection, assessment of sample quality, and data interpretation for effective ASFV surveillance and control. Full article

The COVID-19 pandemic transformed healthcare education, increasing the shift to digital tools and establishing a hybrid curriculum blending online learning with traditional clinical practice. This study aims to understand how this shift impacts the educational growth and skill building of nursing students. A qualitative approach was conducted to understand the experience of Hong Kong nursing students adapting to online learning during the pandemic and beyond. Fifty nursing students were interviewed, and Colaizzi’s phenomenological method revealed key themes in their learning narratives. The analysis revealed four distinct themes characterizing the students’ experiences: (i) Learning on their terms: the mandated shift in healthcare reflecting a lack of agency during the educational transition; (ii) Knowledge without touch: the perceived incompetence of the COVID-19 nursing cohort, highlighting anxieties regarding a lack of hands-on clinical proficiency; (iii) Words left unsaid: The weight of insecurity, indicating a decline in interpersonal skills due to isolation; and (iv) Beyond the perfect algorithm: the unrehearsed art of care, describing the difficulty in translating digital simulations to complex, human-centric patient care. Findings show that while digital progress ensured continuity in education, it also contributed to reduced clinical confidence, weaker communication skills, and shifts in how nursing students approached their learning. Consequently, the post-COVID environment demands that training programs evolve to address these specific deficits. Advancing the existing pandemic-era nursing literature, this study emphasizes the need for diverse, targeted teaching methods to mitigate these gaps. By intentionally bridging theoretical knowledge with hands-on clinical practice, educators can better support student wellbeing and help restore the confidence and competence required of future graduates. Full article

Background: This study evaluated the inter-method agreement of an in-house qualitative CMV real-time PCR assay for the detection of cytomegalovirus (CMV) DNA in various non-plasma clinical specimen types, in comparison with a commercially available comparator assay. Methods: In this prospective comparative study, 186 clinical specimens—including bronchoalveolar lavage fluid (BALF), stool, urine, colonoscopic biopsy, amniotic fluid, and intraocular fluid—were analyzed. A total of 166 samples with valid results from both test systems were included in the inter-method comparison. CMV DNA was detected using the in-house qualitative PCR assay in parallel with the comparator assay (artus^® CMV QS-RGQ kit). Agreement was assessed using positive percent agreement (PPA), negative percent agreement (NPA), overall percent agreement (OPA), and Cohen’s kappa coefficient (κ), in accordance with CLSI EP12-A2 recommendations. Results: Substantial overall inter-method agreement was observed when all specimens were evaluated collectively (κ = 0.66). Agreement metrics were highest in stool, urine, and invasive specimens, whereas BALF samples demonstrated comparatively lower agreement, reflecting potential matrix-related analytical variability. Conclusion: The laboratory-developed qualitative CMV PCR assay demonstrated substantial inter-method agreement with the comparator assay across multiple non-plasma specimen types. The findings highlight specimen-specific variability in qualitative CMV DNA detection and represent analytical concordance between two molecular assays rather than definitive clinical diagnostic accuracy or viral load quantification. Full article

Increased intestinal permeability has been implicated in metabolic dysfunction-associated steatotic liver disease (MASLD), but its relationship with liver fibrosis independent of metabolic risk factors remains unclear. The aim of this study was to investigate the relationship between markers of gut-derived immune activation and liver fibrosis in individuals with metabolic disease. We enrolled 139 adults (48.8 ± 11 years; BMI 33.7 ± 9.5 kg/m²; 50% type 2 diabetes); liver steatosis and fibrosis were estimated using the Hepatic Steatosis Index (HSI) and Fibrotic NASH Index (FNI); liver biopsies were available in a bariatric subgroup. Plasma soluble CD14 (sCD14) and lipopolysaccharide-binding protein (LBP) levels were measured by ELISA kits, and the LBP/sCD14 ratio was calculated. MASLD was present in 78% of participants; in these individuals, sCD14 levels correlated with HSI and FNI (both p < 0.01). In multivariable analysis adjusting for age, sex, BMI, waist circumference, and type 2 diabetes, sCD14 was independently associated with advanced fibrosis (OR: 3.16, 95% CI 1.32–7.55; p = 0.010). This association was confirmed by histology (p = 0.02). Overall, these findings point to a link between gut-derived immune activation and fibrotic burden in MASLD and provide insight into the pathophysiological relevance of the gut–liver axis in metabolic disease. Full article

Background/Objectives: Group A Streptococcus (GAS) pharyngitis is a frequent cause of morbidity in pediatric populations, which requires timely identification to prevent complications such as acute rheumatic fever. Rapid antigen detection tests (RADTs) are practical alternatives to throat culture. This study evaluates the diagnostic performance of the RapidFor™ Strep A test. Methods: This prospective clinical study enrolled 389 pediatric patients aged < 18 years with symptoms suggestive of streptococcal pharyngitis. Two throat swabs were collected from each patient: one for rapid antigen testing with RapidFor™ Strep A and one for culture. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated. Results: Throat culture was positive in 95 of 389 patients (24.4%). The RapidFor™ Strep A test demonstrated a sensitivity of 98.95% (95% confidence interval [CI]: 94.28–99.81%) and a specificity of 96.26% (95% CI: 93.43–97.90%). The PPV was 89.52%, and the NPV was 99.65%. Agreement with culture was excellent (κ = 0.919); in particular, false-positive results accounted for 2.8% and false-negative results accounted for 1.05%. Fever was the strongest clinical indicator associated with positive results. Conclusions: The RapidFor™ Strep A test showed very high diagnostic accuracy compared with throat culture, including an excellent NPV (99.6%), which supports its reliability for ruling out GAS pharyngitis in pediatric settings. The test is an effective screening tool that facilitates timely antibiotic therapy. Full article

Compression textiles have been widely applied in medical, sportswear, and daily usage, with single-jersey structures produced by circular knitting dominating the market due to their thinness and light weight. However, the presence of seams may compromise compression performance and wearer comfort. This study investigates the effects of yarn type, number of yarns, and loop length on pressure, stretchability, and thermal comfort of seamless punch-lace knitted fabrics and explores their potential application in compression textiles. The results show that yarn number is the dominant factor influencing fabric stiffness, stretchability, and pressure. Fabrics with increased yarn content demonstrate higher maximum load and compression pressure. Smaller loop lengths and additional reinforcing yarns improve dimensional stability and resistance to extension. Air permeability decreases with increasing yarn number due to increased fabric thickness and reduced porosity, while thermal conductivity increases and is positively associated with ventilation resistance, indicating a trade-off between heat transfer and breathability. Surface friction and roughness are significantly affected by yarn number, yarn type, and loop length, whereas water vapour permeability shows no significant relationship with the investigated variables. Overall, seamless punch-lace knitted fabrics demonstrate strong potential for compression applications, although careful design is required to balance breathability and thermal comfort. Full article

Objective: To evaluate the utility of synovial iron quantification using Magnetic resonance imaging (MRI) in assessing structural joint damage in the knee of patients with rheumatoid arthritis (RA). Methods: This cross-sectional study employed a two-stage design. In the initial comparative stage, 6 patients with RA and 5 patients with osteoarthritis (OA) were recruited to compare synovial R2* values, a metric derived from iterative decomposition of water and fat with echo asymmetry and least-squares estimation quantitation (IDEAL-IQ) MRI sequences representing synovial iron content. Following this, the RA cohort was expanded to a total of 51 patients to investigate the association between R2* values and clinical parameters, including disease activity and bone erosion. Synovial fluid iron levels were measured with an Iron Assay Kit and synovial iron deposits were semi-quantified via Prussian blue staining. Associations between R2* and clinical and laboratory parameters, including inflammatory factors and joint damage indices, were analyzed using Spearman’s rank correlation. Univariate and multivariate ordered logistic regression models were employed to identify factors associated with bone erosion severity. An R2*-based nomogram was developed and validated using receiver operating characteristic (ROC) analysis and calibration curves. Results: Synovial R2* values were significantly higher in RA patients than those with osteoarthritis (53.66 S⁻¹ vs. 31.38 S⁻¹, p < 0.05), consistent with Prussian blue staining results. While synovial R2* values showed no significant correlation with systemic iron metabolic markers, inflammatory indicators, or the Disease Activity Score 28, they were positively correlated with bone erosion severity (ρ = 0.500, p < 0.001) and negatively associated with the joint space width (ρ = −0.307, p < 0.05). Multivariate analysis identified R2* as an independent indicator linked to bone erosion extent (OR = 2358.336, p < 0.001). The R2*-based nomogram demonstrated good discriminative performance. (AUC = 0.83). Conclusions: The R2* value derived from IDEAL-IQ MRI is a reliable tool for quantifying synovial iron and may represent a promising non-invasive imaging biomarker reflecting bone erosion in RA patients. Full article

The packaging sector presents a significant sustainability challenge, particularly due to the prevalence of plastic packaging. There is a growing interest in sustainable packaging alternatives. The main challenge is to develop packaging with comparable and competitive characteristics. In this context, this manuscript aims to evaluate the performance of lignin-based polyurethane applied as a coating on recycled linerboard. Industrial softwood kraft lignin was fully characterized in terms of purity, functional groups (FTIR and ³¹P NMR) and molecular weight (GPC). Aiming at coating applications, the lignin sample was solubilized in dimethyl sulfoxide (DMSO) and used as a polyol substitute in the reaction, replacing polyethylene glycol (PEG) at levels of 70%, 80%, and 90%. Subsequently, hexamethylene diisocyanate (HDI) was added to initiate polyurethane formation. After polymerization, the coating was applied in multiple layers onto the linerboard paper. Regarding water resistance, all applications demonstrated effectiveness. The lignin-based polyurethane coating improved the Cobb1800 with reductions in the range of 1147.4 to 1155.8 g/m² compared to the uncoated paper. Water vapor permeability was reduced by more than 94%. In the evaluation of oil resistance, samples with three layers and 90% lignin replacement performed particularly well, achieving a high value in a kit test for oil and grease (kit test number 12). These results highlight a promising approach to paper-based packaging, with potential applications across a wide range of products. Full article

Natural fibre hybrid composites have gained attention as cleaner alternatives to synthetic glass fibre systems due to their renewable feedstocks and inherent density advantage. However, moisture ingress degrades fibre–matrix integrity and mechanical performance, making durability a critical design constraint. This study systematically investigates the water absorption kinetics and post-immersion mechanical property retention in ramie–flax/epoxy hybrid composites across four fibre loadings (10–40 wt.%), with the ramie-to-flax weight ratio fixed at 1:1 in all formulations. Tensile, flexural, and impact properties were evaluated under dry and saturated conditions; Fickian diffusion kinetics were analysed to quantify moisture transport parameters; and fracture surfaces were examined by SEM. A density-based material efficiency analysis quantified the lightweighting benefit relative to equivalent synthetic glass/epoxy composites. Water absorption increased monotonically with fibre content; all formulations reached equilibrium after approximately 120 h. The 30 wt.% composite achieved dry-state tensile, flexural, and impact strengths of ca.49 MPa, ca.58 MPa, and 2.82 kJ/m² respectively, retaining ca.78%, ca.69%, and ca.82% after full saturation, superior to all other loadings. These results establish 30 wt.% as the optimal fibre loading for moisture-exposed semi-structural applications, supporting the adoption of ramie–flax composites within a cleaner manufacturing framework. Full article

Renewable energy is an important pillar of decarbonization in reducing the impact of climate change. Among the renewable energy sources, solar photovoltaic energy is one of the fastest-growing across West Africa, especially in Côte d’Ivoire. However, its dependence on weather and climate could affect future power system operations. This study aims to quantify how climate change could affect future solar PV potential in Côte d’Ivoire under the RCP8.5 scenario. For this purpose, we used three regional climate model simulations (RCMs) generated by the new high-resolution Coordinated Regional Climate Downscaling Experiment (CORDEX) for the Africa domain (AFR-22). Future changes were computed for two time slices: the near future (2021–2040) and the middle future (2041–2060), relative to the reference period (1986–2005). The performance of the RCMs and their ensemble mean in simulating relevant climate variables was first evaluated with respect to the ERA5 reanalysis and satellite-based (SARAH-2) data during the reference period. Our results indicate that all available RCMs and their ensemble mean reasonably simulate the annual cycle and the spatial patterns features of surface solar radiation, near-air temperature and solar PV potential in Côte d’Ivoire. We also conclude that Côte d’Ivoire is expected to experience a moderate decrease in annual mean solar PV potential during the mid-21st century. The average decrease in solar PV potential over Côte d’Ivoire could range from 0.55% to 2.16% in the near future and from 1.30% to 3.50% during the middle future, according to the considered RCMs. This decline in solar PV potential will be particularly noticeable during the period from June to October in all climatic zones. Overall, these findings provide valuable information for renewable energy planners to ensure the long-term success of solar PV energy projects in the context of climate change in Côte d’Ivoire. Full article

Background/Objectives: There is a clear need for more options to control the progression of breast cancer and prevent the occurrence of breast cancer in minority populations that have a higher rate of mortality due to triple-negative breast cancer (TNBC) subtypes. Prevalent nutraceuticals such as Ashwagandha (also known as the Indian Winter Cherry) have anti-inflammatory and apoptotic capabilities, as well as the ability to inhibit cancer growth. The purpose of this study is to analyze the novel combination of withaferin A (derived from the Indian Winter Cherry and known to have histone deacetylase inhibition capabilities) and sodium butyrate (a short-chain fatty acid produced from the gut microbiome and known to have DNA methyltransferase inhibition capabilities) treatment on breast cancer-derived cell lines. There is a scientific gap of possible causality of decreasing breast cancer progression when treated with sodium butyrate and withaferin A. Methods: Two in vitro cell viability assays were utilized consisting of [MTT (4,5 Dimethylthiazol-2-yl)] and the neutral red assay to analyze the impact of treatment of compounds alone and in combination on breast cancer cells for 72 h. The Highest Single Agent (HSA) combination analysis was utilized to derive combination indexes for our breast cancer cell types. Protein and gene expression was investigated for Class 1 histone deacetylases, de novo DNA methyltransferase, the p65 subunit of NF-κB, and NFκB1. Lastly, DNA methyltransferase enzymatic activity was analyzed via the Epigentek DNMT Activity/Inhibition ELISA Easy Kit. Results: Through the cell viability assay [MTT (4,5 Dimethylthiazol-2-yl)], MCF−7, MDA−MB−231, and MDA−MB−157 cells were found to have a decrease in cell viability due to combinatorial treatment with withaferin A and sodium butyrate. Western blot results depicted a decrease in protein expression levels for DNA methyltransferases due to the administration of 2.5 mM sodium butyrate and 0.2 µM withaferin A alone and in combination for breast cancer cell lines MCF−7, MDA-MB-231, and MDA−MB−157. Additionally, the combination of these two components have successfully inhibited the progression of the NFκB1 gene within analysis through the quantitative polymerase chain reaction (qPCR). Conclusions: The novel combination of withaferin A and sodium butyrate have markedly reduced the progression of breast cancer-derived cell lines for cell viability, epigenetic DNMT gene expression, as well as inhibiting NFκB1 signaling on the gene expression level. Full article

Background/Objectives: Forensic kinship analysis is a rapidly developing practice that uses genetic data to identify unknown persons of interest through their genetic relatives. It can be used to generate new leads in forensic investigations, especially those involving long-term missing persons and unidentified human remains. More recently, the advent of SNP profiling panels designed specifically for forensic use has led to the exploration of kinship analysis using medium-density SNP data. This study aimed to evaluate the extent to which genetic relationships could be inferred using such data, and to assess the performance of different kinship inference methods. Methods: Kinship analysis was performed with both real and simulated profiles using the panel of SNPs contained within the Verogen ForenSeq^® Kintelligence Kit, with a wide range of relationship types and seven types of kinship inference methods. Results: It was determined that kinship inferences were possible out to the fourth degree of kinship, and all inference methods analysed were equally effective when tested using simulated data. However, some variation between methods was observed when they were analysed using real sample data, suggesting that further study is needed using a larger sample size. Conclusions: The results of this study demonstrate that medium-density SNP data is sufficient for extended kinship inference out to the fourth degree, and that several kinship inference methods are suitable for use with the Verogen ForenSeq^® Kintelligence Kit. These findings will support its application in forensic investigations involving the inference of distant genetic relationships. Full article

Hepatitis E virus (HEV) is recognized as one of the leading causes of acute viral hepatitis (AVH) in developing countries, where it is primarily transmitted through the consumption of contaminated food and water. Although often self-limiting, HEV infection poses a significant public health concern, particularly among pregnant women, due to its potential complications. The present study aimed to determine the seroprevalence of HEV infection in asymptomatic antenatal women attending a tertiary care center in South Punjab, Pakistan. A total of 100 asymptomatic pregnant women were screened for anti-HEV antibodies (IgM and IgG) using an ELISA kit (DIA PRO, Italy). The overall seropositivity rate was found to be 12%, indicating prior exposure to HEV infection in this cohort. Specifically, IgG antibodies were detected in 6% of women and IgM antibodies in 5%, while two women showed evidence of both IgG and IgM positivity, suggestive of recent or ongoing infection. Notably, the majority of participants reported reliance on untreated water sources irrespective of educational background, highlighting environmental risk factors. Although HEV is generally self-limiting, these findings underscore the importance of routine serological screening in antenatal populations to prevent adverse pregnancy outcomes. In addition, increased community awareness regarding transmission routes and preventive measures is essential. Given the scarcity of regional data, this study emphasizes the need for larger-scale epidemiological investigations to better understand the burden of HEV in South Punjab, Pakistan. Full article

Background: Forensic DNA phenotyping (FDP) enables the inference of externally visible characteristics (EVCs) and biogeographic ancestry when conventional STR profiling is inconclusive. The ForenSeq™ Imagen kit (107 SNPs) integrates phenotype-, ancestry-, and Y-SNPs markers; however, its performance under partial genotyping conditions has not been systematically evaluated. Methods: Ninety-four samples from a Mexican mestizo population were analyzed using the ForenSeq™ Imagen kit on the MiSeq FGx™ platform. Due to incomplete genotype recovery, 41 samples with >60% locus detection were selected for downstream analyses. Phenotype prediction was performed using the HIrisPlex-S model, and ancestry inference was assessed through principal component analysis. In silico simulations were conducted to evaluate locus-specific dropout effects. Results: Eye color prediction showed both reduced feasibility (68.3%) and lower overall accuracy (56.1%), primarily driven by systematic prediction failure when rs12913832 (HERC2) was absent, although accuracy among successfully predicted samples remained high (82.1%). In contrast, hair and skin color inference remained feasible in >97% and 100% of evaluable samples, respectively; however, classification accuracy was moderate (70% for hair and 61% for skin), improving substantially when allowing adjacent-category concordance (90.2% for skin). Ancestry inference was robust when at least 27 aiSNPs were detected, and Y-SNPs reliably distinguished male and female samples. In silico analyses confirmed the critical contribution of rs12913832 to eye color model operability. Conclusions: FDP performance under partial genotyping reflects a trade-off between prediction feasibility and accuracy and depends on locus-specific integrity rather than overall genotype completeness. The ForenSeq™ Imagen kit shows robustness for ancestry, sex, hair, and skin prediction, although with variable accuracy, whereas eye color inference remains structurally vulnerable to drop out of high-impact variants. Evaluating FDP systems under realistic non-ideal conditions is essential to define their true operational limits and ensure scientifically robust and responsible implementation. Full article