Search Results (330)

Foodborne pathogens remain a major global concern, demanding rapid, accessible, and determination technologies. Conventional methods, such as culture assays and polymerase chain reaction, offer high accuracy but are time-consuming for on-site testing. This study presents a portable, smartphone-assisted dual-mode biosensor that combines colorimetric and photothermal speckle imaging for improved sensitivity in lateral flow assays (LFAs). The prototype device, built using low-cost components ($500), uses a Raspberry Pi for illumination control, image acquisition, and machine learning-based signal analysis. Colorimetric features were derived from normalized RGB intensities, while photothermal responses were obtained from speckle fluctuation metrics during periodic plasmonic heating. Multivariate linear regression, with and without LASSO regularization, was used to predict Salmonella concentrations. The comparison revealed that regularization did not significantly improve predictive accuracy indicating that the unregularized linear model is sufficient and that the extracted features are robust without complex penalization. The fused model achieved the best performance (R² = 0.91) and consistently predicted concentrations down to a limit of detection (LOD) of 10⁴ CFU/mL, which is one order of magnitude improvement of visual and benchtop measurements from previous work. Blind testing confirmed robustness but also revealed difficulty distinguishing between negative and 10³ CFU/mL samples. This work demonstrates a low-cost, field-deployable biosensing platform capable of quantitative pathogen detection, establishing a foundation for the future deployment of smartphone-assisted, machine learning-enabled diagnostic tools for broader monitoring applications. Full article

The article analyses the direction and intensity of changes occurring in agriculture in mountain areas in Europe between 2000 and 2022. For the calculations, the ESA CCI Land Cover global land-use map set was used. This dataset was established by the European Space Agency (ESA) through the classification of satellite images from sources (MERIS, AVHRR, SPOT, PROBA, and Sentinel-3). In the next step, the organisational features and economic performance of farms located in mountain areas of the European Union were determined for the period 2004–2022. For this purpose, data from the European Farms Accountancy Data Network (FADN-FSDN) were used. Subsequently, using Poland as a case study, the capacity of mountain agriculture to implement key environmental interventions under the Common Agricultural Policy (CAP) 2023–2027 was assessed. The results highlight the varying directions and intensity of organisational changes occurring in mountain agriculture across Europe. They also show that farms can operate successfully in these areas, although their economic situation varies between EU countries. The findings indicate the need for further adaptation of CAP instruments to better reflect the ecological and economic conditions of mountain areas. Strengthening support mechanisms for these regions within the current and future CAP is of crucial importance for protecting biodiversity, promoting sustainable land use, and maintaining the socio-environmental functions of rural mountain landscapes. Our study highlights that the CAP for mountain farms should be targeted, long-term, and compensatory, so as to compensate for the naturally unfavorable farming conditions and support their multifunctional role. The most important assumptions of CAP for mountain farms are a fair system of compensatory payments (LFA/ANCs), support for local and high-quality production, income diversification, and investments adapted to mountain conditions. Full article

The Eocene fourth member of the Shahejie formation (Es4x) in the Bonan Depression, Bohai Bay Basin, records syn-rift sedimentation under alternating arid and humid climates. It provides insight into how orbital-scale climatic fluctuations influenced tectonics, facies patterns, and reservoir distribution. This study integrates 406 m of core data, 92 thin sections, 450 km² of 3D seismic data, and multiple geochemical proxies, leading to the recognition of five facies associations (LFA): (1) alluvial fans, (2) braided rivers, (3) floodplain mudstones, (4) fan deltas, and (5) saline lacustrine evaporites. Three major depositional cycles are defined within the Es4x. Seismic reflections, well-log patterns, and thickness trends suggest that these cycles represent fourth-order lake-level fluctuations (0.8–1.1 Myr) rather than short 21-kyr precession rhythms. This implies long-term climate and tectonic modulation, likely linked to eccentricity-scale monsoon variability. Hyperarid phases are marked by Sr/Ba > 4, δ¹⁸O > +4‰, and thick evaporite accumulations. In contrast, Sr/Ba < 1 and δ¹⁸O < −8‰ reflect humid conditions with larger lakes and enhanced fluvial input. During wet periods, rivers produced sand bodies nearly 40 times thicker than in dry intervals. Reservoir quality is highest in braided-river sandstones (LFA 2) with 12%–19% porosity, preserved by chlorite coatings that limit quartz cement. Fan-delta sands (LFA 4) have <8% porosity due to calcite cementation, though fractures (10–50 mm) improve permeability. Floodplain mudstones (LFA 3) and evaporites (LFA 5) act as seals. This work presents a predictive depositional and reservoir model for arid–humid rift systems and highlights braided-river targets as promising exploration zones in climate-sensitive basins worldwide. Full article

African Swine Fever (ASF) control is severely hampered by the reliance on slow, laboratory-bound diagnostics. While rapid, field-deployable lateral flow assays (LFAs) are urgently needed, the comparative performance of key single-antigen targets remains poorly characterized. This study aimed to develop and systematically evaluate the diagnostic performance of three in-house single-antigen LFAs targeting ASF virus P30, P54, and P72, using swine field samples from Thailand, including a panel of 143 quantitative polymerase chain reaction-negative swine serum samples. The performance of each LFA was compared against a commercial multi-antigen (P32/P62/P72) indirect ELISA, which served as the reference standard, classifying 64 samples as positive and 79 as negative. The P72-based LFA demonstrated perfect diagnostic performance (100% sensitivity, 100% specificity) and perfect agreement (κ = 1.0) with the enzyme-linked immunosorbent assay (ELISA). Similarly, the P30 LFA demonstrated high performance (100% sensitivity, 98.7% specificity) with ‘Almost Perfect’ agreement (κ = 0.9859). In contrast, the P54 LFA was unsuitable, achieving 100% sensitivity but unacceptably low specificity (88.6%) due to a high rate of false positives. Overall, the single-antigen P72 and P30 LFAs demonstrated excellent concordance with the multi-antigen ELISA, supporting their reliable for detecting antibodies against ASFV. Although these assays do not replace molecular methods for acute infection detection, they represent valuable complementary tools for serosurveillance. Full article

Mycoplasma hominis (MH) is a prevalent opportunistic pathogen that is strongly associated with a wide range of urogenital tract infections and severe adverse pregnancy outcomes in clinical settings. Current MH detection methods, including microbial culture and qPCR, are time-consuming and rely on complex equipment, making them unsuitable for scenarios requiring rapid or simplified testing. In this study, we developed a visual readout biosensing platform by synergistically integrating recombinase polymerase amplification (RPA), CRISPR/Cas12a-mediated target nucleic acid recognition, and lateral flow biosensors for the rapid, sensitive, and specific identification of MH. The assay specifically targets the MH-specific 16S rRNA gene, achieving a limit of detection as low as 2 copies/reaction of recombinant plasmid containing the target gene with a total assay time of 60 min. Critical reaction parameters, including Cas12a-crRNA molar ratio, volume of RPA amplicon input, and Cas12a cleavage time, were systematically optimized to maximize the biosensor’s response efficiency and detection reliability. The platform exhibited exceptional specificity, with no cross-reactivity observed against common co-occurring urogenital pathogens, and effectively minimized aerosol contamination risks via a rigorous decontamination workflow. Furthermore, this work represents the first documented implementation of a contamination-control protocol for an MH-specific CRISPR-LFA assay. Notably, testing results from 18 clinical samples demonstrated the high specificity of this assay, highlighting its promising potential for clinical application. Full article

Cardiac troponin I (cTnI) is a critical biomarker for the diagnosis of acute myocardial infarction (AMI), but conventional detection methods are often time-consuming and require specialized laboratory equipment. To meet the need for rapid and feasible detection, there is an urgent demand for methods that are fast, specific, and easy to use. In this study, two aptamers (Tro4 and Tro6), which specifically bind to different epitopes of cTnI, were employed to construct a dual-aptamer sandwich system on a lateral flow assay (LFA) strip. The test strip can deliver results within 10 min and shows a detection limit of 11.70 ng·mL⁻¹. It also exhibited excellent stability after storage at room temperature for up to four months. The assay demonstrated high analytical accuracy, as evidenced by recovery rates from spiked serum samples ranging from 95.11% to 103.17%. These results suggest that the proposed aptamer-based LFA is highly suitable for rapid screening of cTnI, especially in point-of-care settings and resource-limited environments. From a diagnostic perspective, this method holds great promise for improving the timely detection and management of AMI and other myocardial injuries. Full article

Bovine coronavirus(BCoV) is a significant pathogen causing substantial economic losses in the cattle industry through increased calf mortality, reduced growth performance, and decreased milk yield. Rapid and accurate diagnostic methods are therefore essential for controlling BCoV transmission. Current diagnostic methods comprise two primary categories: conventional techniques and cutting-edge innovations. Conventional approaches, including molecular methods like RT-PCR/qRT-PCR and immunological assays such as ELISA and neutralization tests, remain the main diagnostic methods. However, they are limited by laboratory dependency as well as the necessary balance between speed and sensitivity. These limitations have promoted the development of innovative methods, including isothermal amplification, CRISPR/Cas systems, droplet digital PCR, and integrated platforms. This review comprehensively analyzes the advantages, limitations, and applications of current diagnostic methods, highlighting integrated platforms such as RPA-CRISPR-LFA and microfluidics-based LFA. These innovations bridge critical performance gaps by enhancing sensitivity and specificity while enabling field application, demonstrating significant potential as next-generation point-of-care diagnostics for managing this economically critical pathogen. Full article

The rapid detection of 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH), a primary cannabis metabolite, is critical for forensic and workplace drug testing. However, conventional immunoassays often lack sensitivity and objectivity. We developed a portable lateral flow immunoassay device with a microfluidic cartridge and fluorescent reader for the semi-quantitative detection of THC-COOH in urine. A test-to-reference fluorescence ratio was employed to mitigate matrix effects and ensure objective results. The device was validated for accuracy, repeatability, and stability using spiked urine samples and compared against validated LC-MS/MS results on 100 authentic urine samples (50 positive and 50 negative). At a cutoff of 20 ng/mL, the device achieved 100% sensitivity and specificity, with repeatability and reproducibility CVs of below 15%. The cutoff index (COI) strongly correlated with LC-MS/MS results (R² = 0.9471). Crucially, this high correlation with hydrolyzed LC-MS/MS data demonstrates that the antibody recognizes both free and glucuronide-conjugated metabolites, validating its reliability without enzymatic pre-treatment. This microfluidic device enables rapid, sensitive on-site THC-COOH detection, featuring automated data management via Wi-Fi connectivity, enhancing its forensic applicability. Full article

Pulmonary cryptococcosis is an invasive fungal infection usually linked to severe immunosuppression, particularly HIV/AIDS, but is increasingly reported in immunocompetent hosts, including those with uncontrolled diabetes mellitus (DM). We describe a 51-year-old woman with poorly controlled type 2 DM and no other immunosuppressive conditions who developed pulmonary cryptococcosis. Diagnosis was made by microscopy, India ink, cryptococcal antigen lateral flow assay (CrAg LFA), and ITS sequencing; culture was negative. Despite treatment with deoxycholate amphotericin B and fluconazole, the patient died 36 days after admission. A systematic literature review (2000–2025) identified 40 cases of pulmonary cryptococcosis, with 17.5% occurring in patients whose only comorbidity was DM. Cryptococcus neoformans was the most frequent species. Non-culture-based methods, especially CrAg detection, were widely used, underscoring their value for rapid and sensitive diagnosis. Pulmonary cryptococcosis should be considered in diabetic patients even without classical immunosuppression. Broader use of non-culture-based diagnostic tools may enable earlier intervention, which is particularly relevant in resource-limited settings such as Honduras. Full article

Objectives: The purpose of this study is to develop and verify a sandwich-type lateral flow immunoassay (LFA) integrated with a smartphone, enabling semi-quantitative 25-hydroxyvitamin D [25(OH)D] measurement including automated image analysis function, thereby establishing a reliable and accessible vitamin D evaluation system for point-of-care (POCT). Methods: A smartphone-based sandwich-type LFA was constructed, and 25(OH)D was measured semi-quantitatively. The system combined a customized test strip with an automatic image acquisition, calibration, and classification module integrated into an application dedicated to a smartphone. Analysis performance, reproducibility, and equivalence between sample types were comprehensively evaluated. Results: The developed analysis achieved a detection range of 5–100 ng/mL, and there were little interference and cross-reactivity for endogenous substances or structurally similar vitamin D derivatives. The image processing algorithm accurately classified the samples into three clinically important categories: deficiency (<20 ng/mL), insufficient (20–30 ng/mL), and sufficient (>30 ng/mL). Cross-platform testing between Android and iOS devices showed excellent reproducibility (r = 0.99, R² = 0.9967). Comparative analysis with the Atellica IM 1600 analyzer showed a high degree of agreement between 97.0% category consensus and κ = 0.951 (r = 0.99, R² ≥ 0.98). Comparative tests between serum and capillary samples also confirmed a 100% classification agreement rate and an overall diagnostic accuracy of 95.5%. Conclusions: This next-generation smartphone integration platform enables rapid, accurate, and semi-quantitative detection of 25(OH)D from fingerstick and serum specimens. By combining the sandwich-type LFA design with computational-based imaging analysis, the system effectively overcomes the major limitations of small-molecule immunoassay and has the potential to be applied to field diagnosis (POCT), decentralized diagnostics, and vitamin D screening in large populations. Full article

Coal gangue (CG) dumped in open-air piles significantly impacts the surrounding soil environment. To investigate the effects of prolonged CG dumping on soil archaeal communities and their ecological functions, we used metagenomic sequencing to analyze soil samples, including control soil area not impacted by CG (CSL), undisturbed control sediment (CST), atmospheric dust fall area (ADF), and leachate flow area (LFA) samples. The results showed that the dominant phylum and genus of archaea were Thaumarchaeota (30.53–93.39%) and Candidatus Nitrosocosmicus (34.44–69.85%) in the different samples. Significant differences were observed in both α- and β-diversity (p < 0.05); archaeal community composition was primarily influenced by total nitrogen (TN), electrical conductivity (EC), Cu, As, and Cd. The contribution rate of As was the largest, about 44.8%. The metabolic functions of archaea were predominantly related to amino acid metabolism, and there were significant variations in carbon and nitrogen metabolic pathways in different areas. The ppdk gene showed considerable variation between ADF and CSL, and Euryarchaeota was the major contributing phylum to carbon fixation. However, for nitrogen metabolism, the gltB gene displayed marked differences, and the phylum of Thaumarchaeota was the major contributor. This study provides a theoretical foundation for land management and sustainable utilization in CG dump areas. Full article

As a major contributor to environmental pollution in recent years, solid waste has become an increasingly significant concern in the realm of sustainable development. Unmanned Aerial Vehicle (UAV) imagery, known for its high spatial resolution, has become a valuable data source for solid waste detection. However, manually interpreting solid waste in UAV images is inefficient, and object detection methods encounter serious challenges due to the patchy distribution, varied textures and colors, and fragmented edges of solid waste. In this study, we proposed an improved instance segmentation approach called Watershed Mask Network for Solid Waste (WMNet-SW) to accurately retrieve solid waste with precise edges from UAV images. This approach combined the well-established Mask R-CNN segmentation framework with the watershed transform edge detection algorithm. The benchmark Mask R-CNN was improved by optimizing the anchor size and Region of Interest (RoI) and integrating a new mask head of Layer Feature Aggregation (LFA) to initially detect solid waste. Subsequently, edges of the detected solid waste were precisely adjusted by overlaying the segments generated by the watershed transform algorithm. Experimental results show that WMNet-SW significantly enhances the performance of Mask R-CNN in solid waste retrieval, increasing the average precision from 36.91% to 58.10%, F1-score from 0.5 to 0.65, and AP from 63.04% to 64.42%. Furthermore, our method efficiently detects the details of solid waste edges, even overcoming the limitations of training Ground Truth (GT). This study provides a solution for retrieving solid waste with precise edges from UAV images, thereby contributing to the protection of the regional environment and ecosystem health. Full article

Lateral Flow Assays (LFAs) are ubiquitous test platforms due to their affordability and simplicity but are often limited by low sensitivity and lack of flow control. The present work demonstrates the combination of LFAs with centrifugal microfluidic platforms that allows for enhancement of LFAs’ sensitivity via the increase in the dwell time of the analyte at the test line as well as by passing a larger sample volume through the LFA strip. The rate of advancement of the liquid front in the radially positioned NC strip is retarded by the centrifugal force generated on spinning disc; therefore, the dwell time of the liquid front above the test line of LFA is increased. Additionally, integrating a waste reservoir enables passive replenishment of additional sample volume increases total probed volume by approximately 20% (from 50 μL to 60 μL). Comprehensive analysis, including COMSOL multiphysics simulation, was performed to deduce the importance of parameters such as channel height (100–300 μm), disc spin rate (0–2000 rpm), and reaction kinetics (fast vs. slow binding kinetics). The analysis was validated by the experimental observation of the slower-reacting CD79b protein on the test strip. For slower-reacting targets like CD79b, fluorescence intensity increased by ~40% compared to the static LFA. A new merit number, TRc (Transport Reaction Constant), is introduced, which refines the traditional Damköhler number (Da) by including the thickness of the liquid layer (such as the height of the microchannel), which affects the final sensitivity of the assays and is designed to reflect the role channel height plays for surface-based assays (in contrast to the bulk assays). Full article

Background: Type 2 diabetes mellitus (DM) is a major cardiovascular risk factor that induces monocyte dysfunction and contributes to their accumulation in atherosclerotic lesions. Monocyte recruitment and accumulation in the tissues contribute to chronic inflammation and are essential to the pathobiology of diabetes-induced atherosclerosis. However, the mechanisms that drive the accumulation of monocytes in the diabetic environment are not clearly understood. Methods: Primary monocytes from type 2 (T2) DM and non-T2DM individuals were isolated using magnet-assisted cell sorting. To examine the influence of a diabetic milieu on monocyte function, monocytes from T2DM patients, db/db mice, or human monocytes subjected to hyperglycaemia were analysed for their responses to pro-atherogenic cytokines using Boyden chamber assays. Furthermore, the interactions of non-diabetic and diabetic monocytes with TNFα-inflamed endothelium were studied using live-cell imaging under physiological flow conditions. RT-qPCR and FACS were used to study the expression of relevant molecules involved in monocyte-endothelium interaction. Results: CD14⁺CD16⁻ monocytes isolated from T2DM patients or monocytes exposed to hyperglycaemic conditions showed reduced chemotactic responses towards atherosclerosis-promoting cytokines, CCL2 and CX3CL1, indicating monocyte dysfunction. Under flow conditions, the transendothelial migration (TEM) capacity of T2DM monocytes was significantly reduced. Even though these monocytes adhered to the endothelial monolayer, only a few transmigrated. Interestingly, the T2DM monocytes and monocytes exposed to hyperglycaemic conditions accumulated in the ablumen following transendothelial migration. The time period in the ablumen of T2DM cells was prolonged, as there was a significant impairment of the reverse transendothelial migration (rTEM). Mechanistically, the T2DM milieu specifically induced the activation of monocyte integrins, Macrophage-1 antigen (Mac-1; integrin αMβ2 consisting of CD11b and CD18), and Lymphocyte function-associated antigen 1 (LFA-1; αLβ2 consisting of CD11a and CD18). Furthermore, elevated levels of CD18 transcripts were detected in T2DM monocytes. Junctional Adhesion Molecule 3 (JAM-3)–MAC-1 interactions are known to impede rTEM and T2DM milieu-potentiated JAM-3 expression in human coronary artery endothelial cells (HCAEC). Finally, the overexpression of JAM-3 on HCAEC was sufficient to completely recapitulate the impaired rTEM phenotype. Conclusions: Our results revealed for the first time that the enhanced T2DM monocyte accumulation in the ablumen is not secondary to the elevated transmigration through the endothelium. Instead, the accumulation of monocytes is due to the direct consequence of a dysfunctional rTEM, potentially due to enhanced JAM3-MAC1 engagement. Our results highlight the importance of restoring the rTEM capacity of monocytes to reduce monocyte accumulation-dependent inflammation induction and atherogenesis in the T2DM environment. Full article

Epoxy resins are valuable in aerospace, electronics, and high-performance industries; however, their inherently low thermal conductivity (TC) limits applications requiring effective heat dissipation. Recent reports suggest that certain liquid crystalline or partially crystalline epoxy formulations can achieve higher TC, even exceeding 1 W/(m·K). To investigate this, 17 epoxy formulations were prepared, including the commonly used diglycidyl ether of bisphenol A (DGEBA) and two custom-synthesized diepoxides: TME4, which contains rigid aromatic ester linkages with a C4 aliphatic spacer, and LCE-DP, featuring rigid imine bonds. Thermal conductivity was measured using four techniques: laser flash analysis (LFA), modified transient plane source (MTPS), time-domain thermoreflectance (TDTR), and displacement thermo-optic phase spectroscopy (D-TOPS). Additionally, small-angle and wide-angle X-ray scattering (SAXS/WAXS) were performed to detect crystalline or liquid crystalline domains. All formulations exhibited TC values ranging from 0.13 to 0.32 W/(m·K). The TME4–DDS systems, previously reported to be near 1 W/(m·K), consistently measured between 0.26 and 0.30 W/(m·K). Thus, under our synthesis and curing conditions, the elevated TC reported in prior studies was not reproduced, and no strong evidence of crystallinity was observed; indications of local ordering did not translate into higher conductivity. Variations in TC among methods often matched or exceeded the gains attributed to mesophase formation. More broadly, evidence for crystallinity in epoxy thermosets appears weak, consistent with the notion that crosslinking suppresses long-range ordering. Full article