Search Results (955)

The high-quality droplet formation in continuous inkjet printing (CIJ) is crucial for precise character deposition on product surfaces. This process, where a piezoelectric transducer perturbs a high-speed ink stream to generate micro-droplets, is highly sensitive to parameters like ink pressure and transducer amplitude. Suboptimal conditions lead to satellite droplet formation and charge transfer issues, adversely affecting print quality and necessitating reliable monitoring. Replacing inefficient manual inspection, this study develops MBSim-YOLO, a deep learning-based method for automated droplet detection. The proposed model enhances the YOLOv8 architecture by integrating MobileNetv3 to reduce computational complexity, a Bidirectional Feature Pyramid Network (BiFPN) for effective multi-scale feature fusion, and a Simple Attention Module (SimAM) to enhance feature representation robustness. A dataset was constructed using images captured by a CCD camera during the droplet ejection process. Experimental results demonstrate that MBSim-YOLO reduces the parameter count by 78.81% compared to the original YOLOv8. At an Intersection over Union (IoU) threshold of 0.5, the model achieved a precision of 98.2%, a recall of 99.1%, and a mean average precision (mAP) of 98.9%. These findings confirm that MBSim-YOLO achieves an optimal balance between high detection accuracy and lightweight performance, offering a viable and efficient solution for real-time, automated quality monitoring in industrial continuous inkjet printing applications. Full article

Parvovirus B19 is a DNA virus. Most parvoviruses infect animals; Parvovirus B19 infects humans. Parvovirus B19 is mainly transmitted through respiratory droplets during close contact, but additional routes such as transmission through contaminated blood products and vertical transmission from mother to fetus have also been documented. Infections occur throughout the year, with a seasonal increase between late winter and early summer. Clinical symptoms depend on age, and on patients’ immune status. Healthy, immunocompetent individuals experience asymptomatic or mild infections including a febrile rash; serious complications rarely appear, such as rheumatoid-like arthritis or acute myocarditis. Clusters of myocarditis cases following Parvovirus B19 infections appeared in a daycare in Thessaloniki in 2024. To molecularly and phylogenetically characterize Parvovirus B19 strains detected during a pediatric outbreak associated with elevated troponin levels and myocarditis in Northern Greece, and to compare these strains with isolates from adult cases with mild symptoms in order to explore potential associations between viral genetic variability and cardiac involvement. MinION sequencing protocol was performed for nine whole blood samples, seven belonging to children with myocarditis, and two to adults presenting mild symptoms. Statistical analysis was performed with QualiMap 2.3 and relevant tools. Phylogenetic analysis identified distinct viral groups originating from the samples investigated. A distinct branch was formed by the reference genome and the ones of the adults’ samples, while samples from children with myocarditis provided discrete branches differing from the reference one. The findings demonstrate a clear association between Parvovirus B19 infection and myocarditis in the pediatric cases analyzed. The detected viral strains, including variants identified in several samples, support the role of Parvovirus B19 as a contributing factor in post-infectious cardiac involvement. Although these results reinforce the clinical relevance of Parvovirus B19 in childhood myocarditis, expanding the sample size would allow for a more robust characterization of circulating strains and confirmation of the observed patterns. Full article

Borealpox virus (Alaskapox virus, BRPV), a neglected zoonotic Orthopoxvirus (OPXV), has been reported only in Alaska, usually causing mild infections. A recent fatal case in the Kenai Peninsula has raised concerns about its public health impact. Like other OPXVs, BRPV diagnosis relies on molecular tools, making species-specific assays essential. In this study, we developed and validated two BRPV-specific molecular assays: a TaqMan-based real-time PCR (rtPCR) and a droplet digital PCR (ddPCR) using the QIAcuity platform. Both target the viral CC-chemokines inhibitor (vCCI) gene, showing high sensitivity (limit of detection ~0.3–0.5 copies/μL), excellent specificity (no cross-reactivity with other OPXVs or rash-causing viruses), and strong reproducibility. While rtPCR is ideal for routine diagnostics, ddPCR offers absolute quantification without standard curves, enhancing the detection of low viral loads. Although the lack of clinical BRPV-positive samples limits full validation, both assays show strong potential for improving BRPV detection, helping to distinguish it from other OPXVs and supporting the early identification of emerging orthopoxvirus threats. Full article

Objectives: This study aims to explore the therapeutic effect and mechanism of stir-roasted deer velvet antler with ghee (ZLR) on Non-Alcoholic Fatty Liver Disease (NAFLD). Methods: This study used proteomics to analyze the protein composition of roasted deer antler velvet. It established a high-fat diet (HFD)-induced NAFLD rat model and evaluated the therapeutic effects of different dosage groups, including liver injury, oxidative stress, glucose metabolism, steatosis, and insulin homeostasis (via fasting glucose tolerance). Transcriptomics explored the mechanism. Gene expression and Western blot detected lipid metabolism-related gene expression. In vivo experiments validated that ZLR-containing serum alleviates NAFLD and reduces reactive oxygen species levels. Results: The results indicated that ZLR could significantly reduce the body weight, liver weight and degree of hepatic steatosis in HFD rats, improve glycolipid metabolism and insulin sensitivity, and alleviate oxidative stress damage. The mechanism involves activating the adenosine monophosphate-activated protein kinase/peroxisome proliferator-activated receptor (AMPK/PPAR) signaling pathway, regulating the expression of lipid metabolism-related genes, promoting fatty acid oxidation, and reducing fat deposition. The results of in vitro experiments show that ZLR-containing serum can effectively reduce lipid droplet production in liver cells and effectively alleviate oxidative stress damage in liver cells. Conclusions: The traditional Chinese medicine processed product ZLR can regulate lipid metabolism in the body and alleviate the degree of NAFLD by activating the AMPK and PPAR signaling pathways. It provides new ideas for the clinical treatment of NAFLD. Full article

Candida albicans is a major fungal pathogen associated with vulvovaginal candidiasis, and rapid, sensitive detection remains challenging, particularly in amplification-free formats. Here, we report NaPddCas, a microfluidic-free, droplet-based CRISPR/Cas12a detection strategy for qualitative identification of Candida albicans DNA. Unlike conventional bulk CRISPR assays, NaPddCas partitions the reaction mixture into vortex-generated polydisperse droplets, enabling spatial confinement of Cas12a activation events and effective suppression of background fluorescence. This compartmentalization substantially enhances detection sensitivity without nucleic acid amplification or microfluidic devices. Using plasmid and genomic DNA templates, NaPddCas achieved reliable detection at concentrations several orders of magnitude lower than bulk CRISPR/Cas12a reactions. The assay further demonstrated high specificity against non-target bacterial and fungal species and was successfully applied to clinical vaginal secretion samples. Importantly, NaPddCas is designed as a qualitative or semi-qualitative droplet-dependent digital detection method rather than a quantitative digital assay. Owing to its simplicity, sensitivity, and amplification-free workflow, NaPddCas represents a practical approach for laboratory-based screening of Candida albicans infections. Full article

An innovative dispersive liquid–liquid microextraction technique utilizing a solid dispersion was established for the quantification of triazine herbicides in environmental water samples. Naturally derived monoterpenoids were utilized as eco-friendly extraction solvents, markedly decreasing the reliance on harmful extraction solvents. A small amount of Pop Rocks candy served as a solid dispersant; the rapid release of carbon dioxide promoted the generation of fine monoterpenoid droplets, effectively replacing conventional hazardous liquid dispersants. The solidification technique of floating organic droplets facilitated the effective phase separation of monoterpenoids from aqueous samples, thereby obviating the need for centrifugation. Triazine herbicides exhibited good linearity within the concentration range of 0.008–0.8 mg/L with correlation coefficients above 0.99 and detection limits of 0.002 mg/L. The proposed method was effectively implemented on surface and groundwater samples, attaining recoveries between 86.4% and 98.0%. Molecular docking analysis suggests a spontaneous binding between the monoterpenoid and triazine herbicides. A comprehensive green assessment utilizing two evaluation tools confirmed the excellent environmental performance of the method. This technique offers superior greenness and simplicity compared with conventional techniques, demonstrating strong potential for application in the environmental analysis of pesticide residues. Full article

Detecting Escherichia coli on food preparation and processing surfaces is critical for ensuring food safety and preventing foodborne illness. This study focuses on detecting E. coli contamination on common food processing surfaces using UV-C fluorescence imaging and deep learning. Four concentrations of E. coli (0, 10⁵, 10⁷, and 10⁸ colony forming units (CFU)/mL) and two egg solutions (white and yolk) were applied to stainless steel and white rubber to simulate realistic contamination with organic interference. For each concentration level, 256 droplets were inoculated in 16 groups, and fluorescence videos were captured. Droplet regions were extracted from the video frames, subdivided into quadrants, and augmented to generate a robust dataset, ensuring 3–4 droplets per sample. Wavelet-based denoising further improved image quality, with Haar wavelets producing the highest Peak Signal-to-Noise Ratio (PSNR) values, up to 51.0 dB on white rubber and 48.2 dB on stainless steel. Using this dataset, multiple deep learning (DL) models, including ConvNeXtBase, EfficientNetV2L, and five YOLO11-cls variants, were trained to classify E. coli concentration levels. Additionally, Eigen-CAM heatmaps were used to visualize model attention to bacterial fluorescence regions. Across four dataset groupings, YOLO11-cls models achieved consistently high performance, with peak test accuracies of 100% on white rubber and 99.60% on stainless steel, even in the presence of egg substances. YOLO11s-cls provided the best balance of accuracy (up to 98.88%) and inference speed (4–5 ms) whilst having a compact size (11 MB), outperforming larger models such as EfficientNetV2L. Classical machine learning models lagged significantly behind, with Random Forest reaching 89.65% accuracy and SVM only 67.62%. Overall, the results highlight the potential of combining UV-C fluorescence imaging with deep learning for rapid and reliable detection of E. coli on stainless steel and rubber conveyor belt surfaces. Additionally, this approach could support the design of effective interventions to remove E. coli from food processing environments. Full article

Salmonella, a major global foodborne pathogen, is a leading cause of salmonellosis. Quantitative detection of Salmonella provides a scientific basis for establishing microbiological criteria and conducting risk assessments. The plate count method remains the primary approach for bacterial quantification, whereas the most probable number (MPN) method is commonly used for detecting low levels of bacterial contamination. However, both methods are time-consuming and labor-intensive. Validated digital polymerase chain reaction (dPCR) techniques are emerging as promising alternatives because they enable rapid, absolute quantification with high specificity and sensitivity. Herein, we developed a novel droplet dPCR (ddPCR) assay for identifying and quantifying Salmonella using invA as the target. The assay demonstrated high specificity and sensitivity, with a limit of quantification of 1.1 × 10² colony-forming units/mL in meat samples. Furthermore, the log₁₀ values obtained via ddPCR and plate counting exhibited a strong linear relationship (R² > 0.99). Mathematical modeling of growth kinetics further confirmed a high correlation between plate count and ddPCR measurements (Pearson correlation coefficient: 0.996; calculated bias factor: 0.88). Collectively, these results indicate that ddPCR is a viable alternative to the MPN method and represents a powerful tool for the quantitative risk assessment of food safety. Full article

Background: Urine cytology remains widely used for surveillance of non-muscle-invasive bladder cancer despite well-known limitations in sensitivity, especially for low-grade tumors. Uromonitor^®, a molecular assay detecting TERT promoter, FGFR3, and KRAS mutations in voided urine, has emerged as a promising adjunct. To evaluate its suitability for routine use, a consolidated assessment of diagnostic performance and a direct comparison with urine cytology are needed. Methods: We conducted a prospectively registered systematic review (PROSPERO CRD420251173244), synthesizing all available studies that evaluated Uromonitor^® for the detection of urothelial carcinoma of the bladder (UCB). Methodological quality was assessed using the QUADAS-2 framework, and certainty of evidence was evaluated following GRADE for diagnostic tests. Sensitivity was prespecified as the primary endpoint. Comparative datasets were identified, and random-effects meta-analyses were performed for sensitivity, specificity, accuracy, and predictive values (PVs). Results: Across eight cohorts evaluating Uromonitor^®, 832 of 3196 patients (26.0%) had histologically confirmed UCB. Aggregated sensitivity was 0.55 (95% CI 0.52–0.58). Specificity was 0.95 (0.94–0.96). Accuracy was 0.85 (0.83–0.86). PPV was 0.79 (0.76–0.82), and NPV was 0.86 (0.84–0.87). Across seven paired datasets, urine cytology demonstrated a sensitivity of 0.42, a specificity of 0.91, an accuracy of 0.78, a PPV of 0.64, and an NPV of 0.81. Pooled odds ratio for sensitivity was 3.16 (0.73–13.76), while diagnostic accuracy yielded 1.71 (1.01–2.90). Differences in specificity and NPV were not statistically significant, whereas the PPV favored Uromonitor^®, reaching statistical significance in pooled analyses. Conclusions: Uromonitor^® demonstrates higher sensitivity and improved accuracy compared with urine cytology, although current performance remains insufficient for stand-alone surveillance. The sensitivity estimate showed very low certainty due to pronounced heterogeneity, underscoring the need for careful interpretation. With advancing DNA recovery methods, incorporation of droplet digital PCR, and rigorous evaluations in prospective multicenter studies, Uromonitor^® may become an integral element of risk-adapted follow-up strategies. Full article

In semiconductor packaging and microelectronic manufacturing, inkjet printing technology is widely employed in critical processes such as conductive line fabrication and encapsulant dot deposition. However, dynamic printing defects, such as missing droplets and splashing can severely compromise circuit continuity and device reliability. Traditional inspection methods struggle to detect such subtle and low-contrast defects. To address this challenge, we propose MCHB-DETR, a novel lightweight defect detection framework based on RT-DETR, aimed at improving product yield in inkjet printing for semiconductor packaging. MCHB-DETR features a lightweight backbone with enhanced multi-level feature extraction capabilities and a hybrid encoder designed to improve cross-scale and multi-frequency feature fusion. Experimental results on our inkjet dataset show a 29.1% reduction in parameters and a 36.7% reduction in FLOPs, along with improvements of 3.1% in mAP@50 and 3.5% in mAP@50:95. These results demonstrate its superior detection performance while maintaining efficient inference, highlighting its strong potential for enhancing yield in semiconductor packaging. Full article

Background/Objectives: Itaconic acid (ITA) is an immunometabolite with anti-inflammatory and metabolic regulatory functions, but its cellular source and role in brown adipose tissue (BAT) remain unclear. This study aims to reveal the expression patterns of the key ITA synthesis gene Irg1 in BAT at different developmental stages and to investigate the effects of cold exposure and exogenous ITA on BAT metabolic function and cardioprotection. Methods: Single-cell RNA sequencing was used to analyze the gene expression profiles of stromal vascular fraction (SVF) cells in BAT from P7 neonatal and adult mice. Bioinformatic methods were applied to identify cell types expressing Irg1. Cold exposure (4 °C) and exogenous ITA treatment were employed to evaluate BAT morphology, and the ITA content in BAT was detected using gas chromatography–triple quadrupole mass spectrometry, UCP1 protein expression, and body temperature changes. A transverse aortic constriction (TAC) surgery model was established to induce cardiac dysfunction, and BAT excision was performed to explore the BAT-dependent effects of ITA on myocardial hypertrophy, fibrosis, and cardiac function. Results: In P7 neonatal mouse BAT, Irg1 was predominantly expressed in a subset of interferon-responsive activated macrophages (macrophage27), while in adult mice, it was mainly expressed in neutrophils and a functionally similar macrophage subset (macrophage25). Cold exposure significantly suppressed Irg1 expression in neutrophils but did not affect its expression in macrophages, also resulting in a significant decrease in ITA content in BAT. Exogenous ITA significantly enhanced BAT thermogenesis under cold conditions, which manifested as reduced lipid droplets, upregulated UCP1 expression, and increased body temperature. In the TAC model, ITA treatment markedly improved cardiac function, attenuated myocardial hypertrophy and fibrosis, and these protective effects were significantly diminished after BAT excision. Conclusions: ITA promotes cold adaptation and ameliorates cardiac injury by enhancing BAT metabolic function, and its effects depend on the presence of BAT. This study provides new insights for the treatment of metabolic cardiovascular diseases. Full article

Legionnaires’ disease remains a relevant public health concern, with transmission linked to droplets from diverse aquatic environments, and its burden across Europe, including in Portugal, has been trending up. Vehicle windshield washer reservoirs have been proposed as potential, yet underexplored, habitats for Legionella spp. In this study, we investigated 62 windshield washer fluid samples collected in central Portugal. Cultivation on selective BCYE agar supplemented with GVPC and subsequent molecular identification revealed no evidence of Legionella spp. However, 23 morphologically distinct bacterial isolates were recovered, and sequencing confirmed diverse taxa from the genera Brevundimonas, Sphingomonas, Ralstonia, and Xanthobacter. These findings indicate that washer reservoirs can sustain microbial communities characterized by environmental resilience and biofilm-forming potential, traits that overlap with ecological niches exploited by Legionella. Although no Legionella was detected, this work represents the first systematic survey of windshield washer reservoirs in Portugal, emphasizing their potential role as overlooked microbial ecosystems and highlighting the importance of continued surveillance. Broader characterization of microbial communities in such artificial aquatic systems may yield insights into microbial interactions that shape pathogen persistence and suppression. Full article

Chicken respiratory diseases represent multifactorial conditions resulting from viral, bacterial, mycoplasmal pathogens, and environmental factors, causing significant economic losses within the poultry industry. A specific respiratory disease characterized by breathing difficulties and bronchial occlusion due to caseous exudates is termed chicken bronchial obstruction. However, the absence of rapid, precise, and highly sensitive diagnostic methods for differentiation of primary respiratory disease pathogens or opportunistic pathogens, including avian influenza virus (AIV), infectious bronchitis virus (IBV), Pseudomonas aeruginosa (P. aeruginosa), and Escherichia coli (E. coli), constitutes a substantial challenge. This study developed a quadruplex droplet digital polymerase chain reaction (ddPCR) method that targeted the HA gene of H9 subtype AIV, the M gene of IBV, the Pal gene of P. aeruginosa, and the UidA gene of E. coli. Following the optimization of annealing temperature, sensitivity, and repeatability, the minimum detectable concentrations were determined as 3.02 copies/μL for the HA gene of H9 subtype AIV, 3.08 copies/μL for the M gene of IBV, 3.19 copies/μL for the Pal gene of P. aeruginosa, 3.39 copies/μL for the UidA gene of E. coli. No cross-reactivity was observed with Newcastle disease virus (NDV), H5 subtype AIV, H7 subtype AIV, fowl adenovirus serotype 4 (FAdV-4), infectious laryngotracheitis virus (ILTV), Avibacterium paragallinarum, Streptococcus, Salmonella, Pasteurella multocida, and Staphylococcus aureus. The method demonstrated excellent repeatability, with a coefficient of variation (CV) below 9%. The 185 clinical samples collected in Hebei Province China are tested by both quadruplex ddPCR and quadruplex qPCR method and the results compared. The sensitivity of the quadruplex ddPCR method (57.30%; 106/185) slightly exceeded that of the quadruplex qPCR method (49.73%; 92/185). Pathogens or opportunistic pathogens positive rates obtained via the quadruplex ddPCR were 40.00% for H9 subtype AIV, 33.51% for IBV, 24.32% for P. aeruginosa, and 27.57% for E. coli. In comparison, the positive rates of H9 subtypes AIV, IBV, P. aeruginosa, and E. coli from the quadruplex qPCR were 36.22%, 30.81%, 21.62%, and 24.32%, respectively. The coincidence rates between the two methods were 96.22% for H9 AIV, 97.30% for IBV, 97.30% for P. aeruginosa, and 96.76% for E. coli. These results demonstrated that the quadruplex ddPCR method represented a highly sensitive, specific, and rapid technique for identifying H9 subtype AIV, IBV, P. aeruginosa, and E. coli. Full article

Electrokinetics has established itself as a central pillar in microfluidic research, offering a powerful, non-mechanical means to manipulate fluids and analytes. Mechanisms such as electroosmotic flow (EOF), electrophoresis (EP), and dielectrophoresis (DEP) re-main central to the field, once more layers of complexity emerge heterogeneous interfaces, viscoelastic liquids, or anisotropic droplets are introduced. Five research directions have become prominent. Field-driven manipulation of droplets and emulsions—most strikingly Janus droplets—demonstrates how asymmetric interfacial structures generate unconventional transport modes. Electrokinetic injection techniques follow as a second focus, because sharply defined sample plugs are essential for high-resolution separations and for maintaining analytical accuracy. Control of EOF is then framed as an integrated design challenge that involves tuning surface chemistry, engineering zeta potential, implementing nanoscale patterning, and navigating non-Newtonian flow behavior. Next, electrokinetic instabilities and electrically driven micromixing are examined through the lens of vortex-mediated perturbations that break diffusion limits in low-Reynolds-number flows. Finally, electrokinetic enrichment strategies—ranging from ion concentration polarization focusing to stacking-based preconcentration—demonstrate how trace analytes can be selectively accumulated to achieve detection sensitivity. Ultimately, electrokinetics is converging towards sophisticated integrated platforms and hybrid powering schemes, promising to expand microfluidic capabilities into previously inaccessible domains for analytical chemistry and diagnostics. Full article

Norovirus is the leading cause of acute gastroenteritis worldwide, responsible for up to 90% of viral gastroenteritis outbreaks and an estimated 10.6 billion USD in annual economic losses in the U.S. Despite its well-documented seasonality, wastewater surveillance in the Eastern Upper Peninsula of Michigan reveals persistent GI norovirus detection year-round, diverging from national clinical trends that consistently show far greater GII prevalence. To characterize norovirus dynamics in this region, 250 mL wastewater influent grab samples were collected once per week across 14 sites, concentrated using a PEG-based method, and analyzed via digital droplet PCR (ddPCR) for GI and GII concentrations. Across the study period, the rate of positive sites per month ranged from 57 to 100% for GI and 74 to 97% for GII, with mean positivity rates of 85.4% (GI) and 88.7% (GII), indicating that both genogroups were detected frequently at comparable levels. GI was more prevalent in winter and spring (December–May), whereas GII was more prevalent during spring and summer (March–August). Mean GI gene copies per 100 mL ranged from 12,898 (October) to 532,792 (February), while mean GII concentrations ranged from 29,806 (December) to 1,100,215 (May). These patterns contrast with national clinical data, where GI contributes to a small minority of reported norovirus cases. This study explores potential environmental and behavioral factors contributing to this regional pattern. GI norovirus demonstrates greater resistance to wastewater treatment and environmental stability, which may facilitate its persistence in the region. Additionally, congregate living settings, such as college campuses and correctional facilities, may contribute to sustained GI prevalence through foodborne transmission and asymptomatic viral shedding. Overall, these findings suggest that environmental and social factors influence norovirus seasonality and genogroup distribution in this region, underscoring the need for improved monitoring and expanded multi-site wastewater and epidemiological research to better understand norovirus persistence in similar communities. Full article