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The isolation and subsequent molecular analysis of extracellular vesicles (EVs) derived from patient samples is a widely used strategy to understand vesicle biology and to facilitate biomarker discovery. Expressed prostatic secretions in urine are a tumor proximal fluid that has received significant attention as a source of potential prostate cancer (PCa) biomarkers for use in liquid biopsy protocols. Standard EV isolation methods like differential ultracentrifugation (dUC) co-isolate protein contaminants that mask lower-abundance proteins in typical mass spectrometry (MS) protocols. Further complicating the analysis of expressed prostatic secretions, uromodulin, also known as Tamm-Horsfall protein (THP), is present at high concentrations in urine. THP can form polymers that entrap EVs during purification, reducing yield. Disruption of THP polymer networks with dithiothreitol (DTT) can release trapped EVs, but smaller THP fibres co-isolate with EVs during subsequent ultracentrifugation. To resolve these challenges, we describe here a dUC method that incorporates THP polymer reduction and alkaline washing to improve EV isolation and deplete both THP and other common protein contaminants. When applied to human expressed prostatic secretions in urine, we achieved relative enrichment of known prostate and prostate cancer-associated EV-resident proteins. Our approach provides a promising strategy for global proteomic analyses of urinary EVs.

为探索食用葵花籽清选、分级、选种、定价的理论依据，完善建立葵花籽的离散元法分析模型，对食用葵花籽的主要尺寸及均齐性进行了分析。选择内蒙古自治区主栽的3个品种食用葵花籽（1013、SH361、SH363）为研究对象，使用统计学方法，以籽粒的三轴尺寸(长度、宽度、厚度)、平均径（算术平均径、几何平均径）、百粒重为指标，分析葵花籽三轴尺寸的统计分布特性、均齐性以及平均径与百粒重的关系，并按宽度对葵花籽进行了清选分级试验。结果表明：不同品种葵花籽的三轴尺寸均服从正态分布并存在线性相关，且总体上存在显著性差异；葵花籽的均齐性存在差异且其几何平均径、算术平均径与百粒重的变化趋势一致；1013品种选用圆孔筛10 mm和8 mm规格，SH361和SH363品种选用圆孔筛10 mm和8.5 mm规格时，分级效果满足要求。In order to explore the theoretical basis for cleaning, grading, selection, pricing of sunflower seeds, and improve the discrete component analysis model of sunflower seeds, the main dimensions and homogeneity of edible sunflower seeds were analyzed. The three typical edible sunflower seeds (1013, SH361, SH363) planted in Inner Mongolia Autonomous Region were used as research objects, and the statistical distribution characteristics of the three-axis sizes, homogeneity and relationship between average diameter and hundred-grain weight of the sunflower seeds were analyzed with the three-axis size(length, width and thickness), mean size(geometric mean diameter, arithmetic mean diameter) and hundred-grain weight as indexes. The width grading test of sunflower seeds was carried out. The results showed that the three-axis sizes of different varieties of sunflower seeds followed a normal distribution and were linearly correlated with significant differences in general. The homogeneity of sunflower seeds was different, and its geometric mean diameter, arithmetic mean diameter and hundred-grain weight had the same changing trend. When round hole sieves of 10 mm and 8 mm were used for 1013, and round hole sieves of 10 mm and 8.5 mm were used for SH361 and SH363, the grading effect could meet the requirement.

Gallic acid (GA) is a common phenolic acid that is widely used as an antibacterial agent in the food industry. In order to increase its applicability, this study investigated the antibacterial effects and mechanisms of GA on Escherichia coli O157:H7 (E. coli O157:H7) after UV-C light treatment combined with photodynamic therapy. This study evaluated the photodynamic effects of GA after UV-C irradiation (UVC-GA) based on changes in substances and ROS generation. Using E. coli O157:H7 as a model bacterium, the antibacterial effects were evaluated from various aspects, including MIC, MBC, time-kill curve, cell membrane damage, DNA damage, protein damage, motility changes, biofilm formation, and activity. In addition, the antibacterial mechanisms were further investigated by combining RT-qPCR analysis of gene expression. The results of this study demonstrated that GA has the ability to exert photodynamic effects, even after the removal of the UV-C light source, and can effectively reduce the number of viable E. coli O157:H7 cells. Moreover, UVC-GA caused significant damage to the cell membrane, DNA, and proteins, and according to the RT-qPCR results, the antibacterial mechanisms of UVC-GA and GA on E. coli O157:H7 were completely different. Based on the above results, food-sterilizing detergents can be developed and applied to contaminated oyster to provide an effective food cleaning effect. [ABSTRACT FROM AUTHOR]

Background: For patients with congenital heart disease-related pulmonary arterial hypertension (CHD-PAH), cardiopulmonary exercise testing (CPET) can reflect cardiopulmonary reserve function. However, CPET may not be readily accessible for patients with high-risk conditions or limited mobility due to disability. Echocardiography, on the other hand, serves as a widely available diagnostic tool for all CHD-PAH patients. This study was aimed to identify the parameters of echocardiography that could serve as indicators of cardiopulmonary function and exercise capacity. Methods: A cohort of 70 patients contributed a total of 110 paired echocardiogram and CPET results to this study, with 1 year interval for repeated examinations. Echocardiography and exercise testing were conducted following standardized procedures, and the data were collected together with clinically relevant indicators for subsequent statistical analysis. Demographic comparisons were performed using t-tests and chi-square tests. Univariate and multivariate analyses were conducted to identify potential predictors of peak oxygen uptake (peak VO2) and the carbon dioxide ventilation equivalent slope (VE/VCO2 slope). Receiver operating characteristic (ROC) analysis was used to assess the performance of the parameters. Results: The ratio of tricuspid annular plane systolic excursion to pulmonary artery systolic pressure (TAPSE/PASP) was found to be the only independent indicator significantly associated with both peak VO2 and VE/VCO2 slope (both p < 0.05). Additionally, left ventricular ejection fraction (LVEF) and right ventricular fractional area change (FAC) were independently correlated with the VE/VCO2 slope (both p < 0.05). TAPSE/PASP showed the highest area under the ROC curve (AUC) for predicting both a peak VO2 ≤ 15 mL/kg/min and a VE/VCO2 slope ≥ 36 (AUC = 0.91, AUC = 0.90, respectively). The sensitivity and specificity of TAPSE/PASP at the optimal threshold exceeded 0.85 for both parameters. Conclusions: TAPSE/PASP may be a feasible echocardiographic indicator for evaluating exercise tolerance. [ABSTRACT FROM AUTHOR]

Urine is a complex biofluid that reflects both overall physiologic state and the state of the genitourinary tissues through which it passes. It contains both secreted proteins and proteins encapsulated in tissue-derived extracellular vesicles (EVs). To understand the population variability and clinical utility of urine, we quantified the secreted and EV proteomes from 190 men, including a subset with prostate cancer. We demonstrate that a simple protocol enriches prostatic proteins in urine. Secreted and EV proteins arise from different subcellular compartments. Urinary EVs are faithful surrogates of tissue proteomes, but secreted proteins in urine or cell line EVs are not. The urinary proteome is longitudinally stable over several years. It can accurately and non-invasively distinguish malignant from benign prostatic lesions and can risk-stratify prostate tumors. This resource quantifies the complexity of the urinary proteome and reveals the synergistic value of secreted and EV proteomes for translational and biomarker studies. Urine can be used to easily analyse the health of patients. Here, the authors identify proteins in urinary extracellular vesicles that distinguish prostate cancer from benign lesions. [ABSTRACT FROM AUTHOR]

Background: Studies have shown that oxidative stress and its resistance plays important roles in the process of tumor metastasis, and mitochondrial dysfunction caused by mitochondrial DNA (mtDNA) damage is an important molecular event in oxidative stress. In lung cancer, the normal fibroblasts (NFs) are activated as cancer-associated fibroblasts (CAFs), and act in the realms of the tumor microenvironment (TME) with consequences for tumor growth and metastasis. However, its activation mechanism and whether it participates in tumor metastasis through antioxidative stress remain unclear. Methods: The role and signaling pathways of tumor cell derived extracellular vesicles (EVs) activating NFs and the characteristic of induced CAFs (iCAFs) were measured by the transmission electron microscopy, nanoparticle tracking analysis, immunofluorescence, collagen contraction assay, quantitative PCR, immunoblotting, luciferase reporter assay and mitochondrial membrane potential detection. Mitochondrial genome and single nucleotide polymorphism sequencing were used to investigate the transport of mtDNA from iCAFs to ρ0 cells, which were tumor cells with mitochondrial dysfunction caused by depletion of mtDNA. Further, the effects of iCAFs on mitochondrial function, growth and metastasis of tumor cells were analysed in co-culture models both in vitro and in vivo, using succinate dehydrogenase, glutathione and oxygen consumption rate measurements, CCK-8 assay, transwell assay, xenotransplantation and metastasis experiments as well as in situ hybridization and immunohistochemistry. Results: Our findings revealed that EVs derived from high-metastatic lung cancer cells packaged miR-1290 that directly targets MT1G, leading to activation of AKT signaling in NFs and inducing NFs conversion to CAFs. The iCAFs exhibit higher levels of autophagy and mitophagy and more mtDNA release, and reactive oxygen species (ROS) could further promote this process. After cocultured with the conditioned medium (CM) of iCAFs, the ρ0 cells may restore its mitochondrial function by acquisition of mtDNA from CAFs, and further promotes tumor metastasis. Conclusions: These results elucidate a novel mechanism that CAFs activated by tumor-derived EVs can promote metastasis by transferring mtDNA and restoring mitochondrial function of tumor cells which result in resistance of oxidative stress, and provide a new therapeutic target for lung cancer metastasis. [ABSTRACT FROM AUTHOR]

Background: Accurately stratifying patients with pulmonary arterial hypertension (PAH) is very important, and traditional risk scores still have internal heterogeneity. This study aimed to construct a risk stratification model that can accurately identify clinical worsening (CW) events in conventional low‐intermediate risk patients with pulmonary hypertension under targeted drug treatment by using echocardiographic parameters. Methods: This study is a single‐center, prospective study, including 105 PAH patients who underwent regular follow‐up at Guangdong Provincial People's Hospital from October 2021 to April 2023. The primary endpoint was the occurrence of CW, including death, hospitalization due to pulmonary hypertension, escalation of targeted drug therapy, and worsening of PAH. The predictive value of the echocardiography‐based three‐strata risk model was assessed using Kaplan‐Meier curves and COX regression analysis. Results: A total of 98 PAH patients were ultimately included in this study. The median follow‐up duration was 26 months (range 7–28 months). The echocardiography‐based three‐strata model included the ratio of tricuspid annular plane systolic excursion and pulmonary artery systolic pressure (TAPSE/PASP) and inferior vena cava (IVC). The echocardiography‐based three‐strata model had higher diagnostic value (C‐index =.76) compared to the 2022 ESC/ERS three‐strata model and four‐strata model (C‐index =.66 and C‐index =.61, respectively). PAH patients with lower TAPSE/PASP and wider IVC showed a higher CW rate compared to patients with higher TAPSE/PASP and normal IVC (HR = 15.1, 95%CI:4.4‐51.9, p <.001). Conclusion: The echocardiography‐based three‐strata model based on TAPSE/PASP and IVC can effectively improve the stratification of low‐intermediate risk PAH patients under targeted treatment. [ABSTRACT FROM AUTHOR]