Search Results (2,560)

Chlorpyrifos CPF (O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate), known also as Chlorpyrifos-ethyl, is one of the most utilized organophosphorus pesticides worldwide. Additionally, CPF could be used as a chemical warfare agent surrogate. Although its acute toxicity is not high, it is responsible for both a large number of intoxications and chronic, delayed neurological effects. In this work, it is reported for the first time the qualitative and quantitative response produced by CPF vapors, using a pocket-held Time-of-Flight Ion Mobility Spectrometer (ToF IMS) with a non-radioactive ionization source and ammonia doping, model LCD-3.2E (Smiths Detection Ltd.), operated near ambient temperature (below 30 °C). Spectra of CPF in positive ion mode included two distinct product ion peaks; thus, identification of CPF vapors by IMS relies on these peaks—the monomer M·NH₄⁺ with reduced ion mobility K₀ = ca. 1.76 cm²V⁻¹s⁻¹ and the dimer M₂·NH₄⁺ with K₀ = ca. 1.47 cm²V⁻¹s⁻¹ (where M may be assignable to CPF molecule)—and positive reactant ions (Pos RIP) have K₀ = ca. 2.25 cm²V⁻¹s⁻¹. Excellent sensitivity, with a limit of detection LOD of 0.72 ppb_v (10.5 μg m⁻³) and a limit of quantification LOQ of 2.41 ppb_v (35.1 μg m⁻³), has been noticed; linear response was up to 100 ppb_v, while saturation occurs over ca. 1000 ppb_v (14.6 mg m⁻³). Our results demonstrate that this method provides a robust tool for both off-site and on-site detecting and quantifying CPF vapors at trace levels, which has strong implications for either industrial hygiene or forensic investigations concerning the pesticide Chlorpyrifos, as well as for monitoring of environmental contamination by organophosphorus pesticides. Full article

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR-associated) systems, originally identified as prokaryotic adaptive immune mechanisms, have rapidly evolved into powerful tools for molecular diagnostics. Leveraging their precise nucleic acid targeting capabilities, CRISPR diagnostics offer rapid, sensitive, and specific detection solutions for a wide array of targets. This review delves into the fundamental principles of various Cas proteins (e.g., Cas9, Cas12a, Cas13a) and their distinct mechanisms of action (cis- and trans-cleavage). It highlights the diverse applications spanning infectious disease surveillance, cancer biomarker detection, and genetic disorder screening, emphasizing key advantages such as speed, high sensitivity, specificity, portability, and cost-effectiveness, particularly for point-of-care (POC) testing in resource-limited settings. The report also addresses current challenges, including sensitivity limitations without pre-amplification, specificity issues, and complex sample preparation, while exploring promising future trajectories like the integration of artificial intelligence (AI) and the development of universal diagnostic platforms to enhance clinical translation. Full article

Background: Gemcitabine (GEM) remains a cornerstone in the treatment of pancreatic cancer. Upon exposure to GEM, pancreatic cancer cells (PCCs) tend to adapt quickly to outcompete drug-induced cytotoxicity, thereby contributing to treatment failure. Thus, understanding GEM-induced molecular changes in PCCs is important. Methods: Three primary PCC lines (PCC-1, PCC-2, PCC-7) and Mia PaCa-2 cultured for 40 passages (p) in the absence (control) or presence of GEM (GemR) were assessed for phenotypic changes. Proteome profiles for all PCCs at p10, p20, p25, p30, p35, and p40 were obtained using mass spectrometry (MS). Protein expression was determined using immunoblotting. Differentially abundant proteins (DAPs) were evaluated for enrichment of functional and biological attributes and protein–protein interactions. Results: GEM sensitivity and growth were both reduced in GemR versus paired controls for all four PCC lines. MS mapped > 7000 proteins in each PCC line, and the abundance of 70–83% of these was found to be significantly altered when comparing all sample groups. Proteomic changes in GemR versus paired controls differed remarkably among the PCCs and were affected by passaging and treatment duration. DAPs at p40 were mostly related to metabolic pathways, including nucleotide metabolism and diverse cell growth processes. Several closely related DAPs and multiple hub proteins in each PCC line were identified. Conclusions: Overall, this study revealed cell-line-specific, heterogeneous changes in proteome profiles of PCCs following their long-term exposure to GEM, and these were likely affected by treatment duration, dosage, and passaging. Full article

Automatic detection of clue cells is crucial for rapid diagnosis of bacterial vaginosis (BV), but existing algorithms suffer from low sensitivity. This is because clue cells are highly similar to normal epithelial cells in terms of macroscopic size and shape. The key difference between clue cells and normal epithelial cells lies in the surface texture and edge morphology. To address this specific problem, we propose an clue cell detection algorithm named CA-YOLO. The contributions of our approach lie in two synergistic and custom-designed feature extraction modules: the context-aware module (CAM) extracts and captures bacterial distribution patterns on the surface of clue cells; and the shuffle global attention mechanism (SGAM) enhances cell edge features and suppresses irrelevant information. In addition, we integrate focal loss into the classification loss to alleviate the severe class imbalance problem inherent in clinical samples. Experimental results show that the proposed CA-YOLO achieves a sensitivity of 0.778, which is 9.2% higher than the baseline model, making the automated BV detection more reliable and feasible. Full article

Background: The rapid emergence of antibiotic resistance in Acinetobacter baumannii has led the World Health Organization (WHO) to designate it as a “high priority” pathogen. The emergence of multidrug-resistant (MDR) and pandrug-resistant (PDR) strains poses considerable treatment challenges. As antimicrobial resistance (AMR) escalates toward a post-antibiotic era, innovative therapeutic solutions are urgently needed. Objectives: To clone, over-express, and characterize a novel endolysin, LysTAC1, from Acinetobacter phage TAC1 for its antibacterial efficacy against multidrug-resistant bacteria. Methods: A 24 kDa endolysin featuring a glycoside hydrolase Family 19 chitinase domain was tested against carbapenem-resistant Acinetobacter baumannii clinical isolates and various Escherichia coli strains following outer membrane permeabilization with Ethylenediaminetetraacetic acid (EDTA). Stability assays and molecular docking studies were performed. Results: LysTAC1 demonstrated potent lytic activity against Gram-negative bacteria but showed no activity against Gram-positive bacteria (Staphylococcus aureus ATCC 29213 and Enterococcus gallinarum HCD 28-1). LysTAC1 maintained activity across pH 6–9 and temperatures 4–65 °C, with differential sensitivity to metal ions where K⁺ showed no inhibitory effect at any concentration (0.1–100 mM), and Fe²⁺ was non-inhibitory at lower concentrations (0.1–1 mM), while Mg²⁺ and Ca²⁺ demonstrated concentration-dependent inhibition across the tested range (0.1–100 mM). Molecular docking revealed LysTAC1 interactions with chitinase substrates 4-nitrophenyl N-acetyl-β-D-glucosaminide and 4-nitrophenyl N, N-Diacetyl-β-D-chitobioside, with binding energies of −5.82 and −6.85 kcal/mol, respectively. Conclusions: LysTAC1 shows significant potential as a targeted therapeutic agent against A. baumannii with robust stability under physiological conditions. Full article

Closed hydroponic systems for strawberries (Fragaria × ananassa Duch.) are infrequently used because the crop is highly sensitive to salt accumulation and prone to root diseases, resulting in yield reduction. This study investigated semi-closed hydroponic systems using various drainage recycling ratios (30%, 50%, and 70% of drainage EC) to determine their impact on yield, fruit quality, and antioxidant properties. Recycling at moderate levels (30–50%) effectively maintained ionic balance, particularly with respect to K/N and K/Ca ratios, which enabled stable yields and increased fruit weight similar to the control (open hydroponic system) group. Conversely, a high recycling ratio (70%) led to ionic imbalances—characterized by increased K/N ratios and higher concentrations of Na⁺, Cl⁻, and SO₄²⁻—that were associated with decreased fruit size. Measures of antioxidant capacity, such as total phenol and flavonoid content, ferric reducing antioxidant power, and DPPH activity, were not significantly influenced by the recycling ratio alone. Nevertheless, the relatively elevated antioxidant activity observed at the 70% recycling level indicates a mild ionic and osmotic stress response likely caused by increased salt concentration. Changes related to the cropping system season, rather than ion variations from recycling, exerted a stronger influence on antioxidant accumulation. In summary, moderate drainage recycling facilitates optimal fruit production without negatively affecting quality, while excessive recycling may increase antioxidant activity but leads to reduced yields. The results provide practical recommendations for optimizing nutrient reuse in semi-closed strawberry hydroponic systems. Full article

Objectives: This study investigated fractional anisotropy (FA) differences within key white matter tracts across patient groups stratified by Montreal Cognitive Assessment (MoCA) scores, aiming to evaluate FA’s potential as a biomarker for cognitive impairment. Methods: Seventy participants (aged 57–96 years) were categorized into high (HP, MoCA ≥ 26), moderate (MP, MoCA 18–25), and low (LP, MoCA < 18) cognitive performance groups. Diffusion Tensor Imaging (DTI) was used to obtain FA values in corticospinal tracts, superior longitudinal fasciculus, inferior fronto-occipital fasciculus, and cingulum. Statistical analyses included ANOVA and post-hoc tests. Results: Significant differences in FA values and normative percentiles were observed across cognitive groups in several tracts. Notably, the MP group exhibited significantly higher FA values in the Left Superior Longitudinal Fasciculus—Arcuate (mean FA 0.329 vs. LP 0.306, p = 0.033) and Right Superior Longitudinal Fasciculus—Arcuate (mean FA 0.329 vs. LP 0.306, p = 0.009), Left Inferior Fronto-Occipital Fasciculus (mean FA 0.308 vs. LP 0.283, p = 0.021), and Right Inferior Fronto-Occipital Fasciculus (mean FA 0.289 vs. LP 0.266, p = 0.017) compared to the LP group. Conclusions: Our findings reveal significant FA alterations across MoCA-defined cognitive groups, with moderate impairment showing higher FA than low performance. This suggests FA may reflect complex microstructural changes in early cognitive decline. While our modest sample size, particularly in the low-performance group, limits definitive conclusions, these results highlight the need for larger, multimodal studies to validate FA’s role as a sensitive, albeit complex, biomarker for cognitive impairment. Full article

Biliary tract diseases, including both benign and malignant conditions such as cholangitis, cholelithiasis, primary sclerosing cholangitis, cholangiocarcinoma, and gallbladder cancer, present significant challenges for timely diagnosis and effective clinical management. Conventional diagnostic approaches, which primarily rely on imaging and standard laboratory tests, often lack the sensitivity and specificity needed for early detection, accurate risk stratification, and personalized treatment planning. In recent years, advancements in point-of-care (POC) diagnostic technologies, along with the identification and validation of novel biomarkers, have begun to reshape the diagnostic landscape. This review provides a comprehensive overview of the clinical utility and limitations of current POC tests and biomarkers, ranging from well-established markers such as carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) to emerging molecular indicators such as circulating microRNAs and circulating tumor DNA. We examine their applications across acute management, chronic disease monitoring, and cancer detection; identify existing gaps in diagnostic practice; and discuss strategies for incorporating these tools into standard clinical workflows to enhance patient outcomes. Full article

This study comprehensively evaluated the antimicrobial performance of copper ions against three bacterial species relevant to water systems: E. coli (ATCC 25922), P. aeruginosa (ATCC 27853), and S. epidermidis (ATCC 12228). Disinfection kinetics were determined at three copper concentrations (0.5, 1.5, and 3.3 mg/L) using the Gard model. E. coli exhibited the highest susceptibility, with inactivation rate constants of 0.63, 3.27, and 9.83, achieving complete inactivation at 3.3 mg/L. P. aeruginosa was the most resistant, showing values below 1.0 across all concentrations, while S. epidermidis displayed intermediate responses. Selected experiments further examined the influence of growth phase, temperature, and water chemistry. Exponential-phase cells were more sensitive than stationary-phase cultures, and higher temperatures (37 °C vs. 5 °C) significantly enhanced inactivation. Moderate bicarbonate (50 mg/L) improved bacterial removal by stabilizing soluble Cu²⁺ ions (2.60 lg reduction), whereas elevated calcium and magnesium (Ca²⁺ 100 mg/L, Mg²⁺ 50 mg/L) reduced effectiveness (≤2.10 lg reduction) through competitive interactions. In addition to culture-based methods, adenosine triphosphate (ATP) bioluminescence assays and flow cytometry (FCM) provided complementary insights, confirming early metabolic disruption and membrane damage prior to culturability loss in selected experiments. Full article

Background: The cerebral compliance (or compensatory reserve) index, RAP, is a critical yet underutilized physiological marker in the management of moderate-to-severe traumatic brain injury (TBI). While RAP offers promise as a continuous bedside metric, its broader cerebral physiological context remains partly understood. This study aims to characterize the burden of impaired RAP in relation to other key components of cerebral physiology. Methods: Archived data from 379 moderate-to-severe TBI patients were analyzed using descriptive and threshold-based methods across three RAP states (impaired, intact/transitional, and exhausted). Agglomerative hierarchical clustering, principal component analysis, and kernel-based clustering were applied to explore multivariate covariance structures. Then, high-frequency temporal analyses, including vector autoregressive integrated moving average impulse response functions (VARIMA IRF), cross-correlation, and Granger causality, were performed to assess dynamic coupling between RAP and other physiological signals. Results: Impaired and exhausted RAP states were associated with elevated intracranial pressure (p = 0.021). Regarding AMP, impaired RAP was associated with elevated levels, while exhausted RAP was associated with reduced pulse amplitude (p = 3.94 × 10⁻⁹). These two RAP states were also associated with compromised autoregulation and diminished perfusion. Clustering analyses consistently grouped RAP with its constituent signals (ICP and AMP), followed by brain oxygenation parameters (brain tissue oxygenation (PbtO₂) and regional cerebral oxygen saturation (rSO₂)). Cerebral autoregulation (CA) indices clustered more closely with RAP under impaired autoregulatory states. Temporal analyses revealed that RAP exhibited comparatively stronger responses to ICP and arterial blood pressure (ABP) at 1-min resolution. Moreover, when comparing ICP-derived and near-infrared spectroscopy (NIRS)-derived CA indices, they clustered more closely to RAP, and RAP demonstrated greater sensitivity to changes in these ICP-derived CA indices in high-frequency temporal analyses. These trends remained consistent at lower temporal resolutions as well. Conclusion: RAP relationships with other parameters remain consistent and differ meaningfully across compliance states. Integrating RAP into patient trajectory modelling and developing predictive frameworks based on these findings across different RAP states can map the evolution of cerebral physiology over time. This approach may improve prognostication and guide individualized interventions in TBI management. Therefore, these findings support RAP’s potential as a valuable metric for bedside monitoring and its prospective role in guiding patient trajectory modeling and interventional studies in TBI. Full article

Chrysotila dentata (Haptophyta), a harmful algal bloom (HAB) species frequently occurring in coastal waters of China, is one with strong environmental adaptability that poses a serious threat to marine ecosystems and fisheries. Current molecular detection techniques and early warning systems for this species remain limited. To address this, we developed a rapid and highly sensitive detection method for C. dentata. This method integrates recombinase polymerase amplification (RPA) with CRISPR-LbaCas12a and lateral flow dipstick (LFD) technologies, enabling visual readout of results. Key parameters, including the single-stranded DNA (ssDNA) reporter concentration, reaction time, and temperature, were systematically optimized. Field water sample testing demonstrated high specificity and sensitivity, achieving a detection limit of 5 × 10⁻⁶ pg μL⁻¹ for genomic DNA under laboratory conditions and 2.82 × 10¹ cells mL⁻¹ in simulated environmental samples. The entire detection process takes only 1 h (at a constant 39 °C), and results can be directly interpreted via LFD strips. For early warning and prevention of C. dentata outbreaks, this assay provides a powerful, reliable, and field-ready monitoring tool. Full article

Hydrogen injection timing (HIT) plays a crucial role in the combustion and emission characteristics of a hydrogen/n-butanol dual-fuel engine with hydrogen direct injection. This study employed an integrated approach combining three-dimensional simulation modeling and engine test bench experiments to investigate the effects of HIT on engine performance. In order to have a more intuitive understanding of the physical and chemical change processes, such as the stratification state and combustion status of hydrogen in the cylinder, and to essentially explore the internal mechanism and fundamental reasons for the improvement in performance of n-butanol engines by hydrogen addition, a numerical study was conducted to examine the effects of HIT on hydrogen stratification and combustion behavior. The simulation results demonstrated that within the investigated range, at 100 °CA BTDC hydrogen injection time, hydrogen forms an ideal hydrogen stratification state in the cylinder; that is, a locally enriched hydrogen zone near the spark plug, while there is a certain distribution of hydrogen in the cylinder. Meanwhile, the combustion state also reaches the optimal level at this hydrogen injection moment. Consequently, 100 °CA BTDC is identified as the optimal HIT for a hydrogen/n-butanol dual-fuel engine. At the same time, an experimental study was performed to capture the actual complex processes and comprehensively evaluate combustion and emission characteristics. The experimental results indicate that both dynamic performance (torque) and combustion characteristics (cylinder pressure, flame development period, etc.) achieve optimal values at the HIT of 100 °CA BTDC. Notably, under lean-burn conditions, the combustion parameters exhibit greater sensitivity to HIT. Regarding emissions, the CO and HC emissions initially decreased slightly, then gradually increased with advanced injection timing. The 100 °CA BTDC timing effectively reduced the CO emissions at λ = 0.9 and 1.0. CO emissions at λ = 1.2, and showed minimal sensitivity to the injection timing variations. Therefore, optimized HIT facilitates enhanced combustion efficiency and emission performance in hydrogen-direct-injection n-butanol engines. Full article

Pine wilt disease, which is induced by pine wood nematode (PWN, Bursaphelenchus xylophilus), has caused huge economic and ecological losses. To overcome the drawbacks of chemical control against PWN, twenty compounds were screened, and a synergistic botanical–chemical combination was identified. A proportion of abamectin to rotenone of 7:3 (5.73 and 1.78 mg/L, respectively) achieved the highest co-toxicity coefficient of 231.09 with a median lethal concentration of 3.18 mg/L. It revealed 0% mortality in Pinus massoniana seedlings at 60 days post-treatment when applied at 400 times the synergistic concentration (2.29 g/L abamectin + 0.71 g/L rotenone) at 7 days after PWN inoculation. Furthermore, the synergistic combination significantly affected the physiological activity and population dynamics of PWN. Female oviposition was reduced by 71.92%, the egg hatching rates declined to 13.09 ± 0.02%, and head thrashing frequency was inhibited by 99.23 ± 0.01%. The enzymatic activities of peroxidase, acetylcholinesterase, succinate dehydrogenase, and glutathione S-transferase were significantly increased, while the population size declined by 96.17%. Transcriptomic and gene expression analyses suggested a potential “Na⁺/Ca²⁺/Cl⁻ ionic storm,” since the synergistic combination significantly activated genes associated with voltage-gated calcium channels, glutamate-gated chloride channels, and amiloride-sensitive sodium channels. These findings provide an eco-friendly strategy for PWN management via chemical control. Full article

Senecavirus A (SVA) is an emerging pathogen responsible for vesicular lesions and neonatal mortality in swine. In the absence of effective vaccines or therapeutics, early and accurate diagnosis is essential for controlling SVA outbreaks. Although nucleic acid-based detection methods are commonly employed, there remains a pressing need for rapid, convenient, highly sensitive, and specific diagnostic tools. Here, we developed a two-pot assay combining recombinase polymerase amplification (RPA) with CRISPR/Cas12a containing crRNA targeting canonical protospacer adjacent motifs (PAMs) for simple, rapid, and visual identification of SVA in clinical samples. Subsequently, we successfully streamlined this system into a one-pot assay by selecting a specially designed crRNA targeting suboptimal PAM and integrating RPA amplification reagents and CRISPR/Cas12a detection components into a single reaction system in one tube. The developed methods exhibited diagnostic specificity, showing no cross-reactivity with four major swine viruses, while showing remarkable sensitivity with a lower detection limit of just two copies. Clinical validation in field samples using these two methods revealed perfect agreement (100% concordance) with conventional quantitative PCR (qPCR) results (sample size, n = 28), with both assays completing detection within 30 min. These results demonstrate that both the one-pot and two-pot RPA-CRISPR/Cas12a assays offer a reliable and efficient method for detecting SVA in this pilot study. Despite the limited sample size, the assays combine rapid reaction time with high sensitivity and specificity, showing great potential for future diagnostic applications. Full article