Dataset supporting paper 'Dietary n-3 polyunsaturated fatty acids alter the number, fatty acid profile and coagulatory activity of circulating and platelet-derived extracellular vesicles'

This dataset includes all reports and summaries of raw data supporting the results presented in the paper "Dietary n-3 polyunsaturated fatty acids alter the number, fatty acid profile and coagulatory activity of circulating and platelet-derived extracellular vesicles". The objective of this paper is to investigate whether daily supplementation of participants at moderate risk of (cardiovascular diseases) CVDs with 1.8 g/d of fish oil-derived n-3 polyunsaturated fatty acids (PUFAs) altered the generation, composition and function of circulating and platelet-derived extracellular vesicles (EVs). Data about EVs parameters include: i. the numbers, and size of circulating total EVs and in vitro generated platelet-derived EVs (PDEVs) measured by Nanoparticle Tracking Analysis (NTA); ii. the numbers of EV subpopulations (i.e. phosphatidylserine-positive EVs (PS+EVs), PDEVs, endothelial-derived EVs (EDEVs)) and PS expression on PDEVs generated in vitro from platelets measured by flow cytometry (FCM); iii. the procoagulatory activity of circulating EVs (in vitro thrombogenic potential in activating tissue factor-dependent thrombin generation) measured by thrombin generation assay. The coagulatory behaviour of PDEVs generated in vitro from platelets measured by thrombin generation, clot formation, fibrinolysis and in vivo thrombus formation assays. Data about conventional cardiovascular risk markers include i. body mass index (BMI) measured by Tanita; ii. blood pressure measured by upper arm blood pressure monitor; iii. plasma lipid profile (i.e. triacylglycerol (TAG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and glucose concentrations measured by iLab. Data about thrombogenic markers include: i. plasma platelet aggregation measured by 96-well high-throughput aggregometry; ii. plasma thrombin generation measured by thrombin generation assay; iii. plasma clot growth and fibrinolysis measured by thrombodynamics analyzer

Huobahuagen tablet improves renal function in diabetic kidney disease: a real-world retrospective cohort study

ObjectiveWe aimed to explore the value of Huobahuagen tablet (HBT) in improving decreased renal function for patients with diabetic kidney disease (DKD) over time.MethodsThis was a single-center, retrospective, real-world study on eligible 122 DKD patients who continued to use HBT + Huangkui capsule (HKC) therapy or HKC therapy without interruption or alteration in Jiangsu Province Hospital of Chinese Medicine from July 2016 to March 2022. The primary observation outcomes included estimated glomerular filtration rate (eGFR) at baseline and 1-, 3-, 6-, 9-, and 12-month follow-up visits and changes in eGFR from baseline (ΔeGFR). Propensity score (PS) and inverse probability treatment weighting (IPTW) were used to control for confounders.ResultseGFR was significantly higher in the HBT + HKC group than in the HKC alone group at the 6-, 9-, and 12-month follow-up visits (p = 0.0448, 0.0002, and 0.0037, respectively), indicating the superiority of HBT + HKC over HBT alone. Furthermore, the ΔeGFR of the HBT + HKC group was significantly higher than that of the HKC alone group at the 6- and 12-month follow-up visits (p = 0.0369 and 0.0267, respectively). In the DKD G4 patients, eGFR was higher in the HBT + HKC group at the 1-, 3-, 6-, 9-, and 12-month follow-up visits compared with baseline, with statistically significant differences at the 1-, 3-, and 6- month follow-up visits (p = 0.0256, 0.0069, and 0.0252, respectively). The fluctuations in ΔeGFR ranged from 2.54 ± 4.34 to 5.01 ± 5.55 ml/min/1.73 m2. Change in the urinary albumin/creatinine ratio from baseline did not exhibit a significant difference between the two groups at any of the follow-up visits (p > 0.05 for all). Adverse event incidence was low in both groups.ConclusionThe findings of this study based on real-world clinical practice indicate that HBT + HKC therapy exhibited better efficacy in improving and protecting renal function with a favorable safety profile than HKC therapy alone. However, further large-scale prospective randomized controlled trials are warranted to confirm these results

Circulating extracellular vesicles are strongly associated with cardiovascular risk markers

Background: Extracellular vesicles (EVs) are submicron membrane-bound vesicles released from various cells, which are emerging as a potential novel biomarker in cardiovascular diseases (CVDs) due to their procoagulatory and prothrombotic properties. However, there is little information about the relationships between circulating EVs and conventional and thrombogenic risk markers of CVDs. Objective: To investigate the relationships between circulating EVs, conventional cardiovascular risk markers and thrombogenic markers in subjects with moderate risk of CVDs. Design: Subjects (n=40) aged 40-70 years with moderate risk of CVDs were recruited and assessed for body mass index, blood pressure and plasma lipid profile, as well as platelet aggregation, clot formation, thrombin generation and fibrinolysis. Numbers of circulating EVs were assessed by Nanoparticle Tracking Analysis and flow cytometry. A range of assays were used to assess the procoagulatory activity of plasma and circulating EVs. Results: Circulating EV numbers were positively associated with body mass index, blood pressure, plasma triacylglycerol concentration and overall CVD risk. Higher circulating EV numbers were also associated with increased thrombin generation and enhanced clot formation, and EVs isolated from subjects with moderate CVD risk promoted thrombin generation ex vivo. Higher numbers of endothelial-derived EVs were associated with a greater tendency for clot lysis. Plasma triacylglycerol concentration and diastolic blood pressure independently predicted circulating EV numbers, and EV numbers independently predicted aspects of thrombin generation and clot formation and 10-year CVD risk. Conclusion: Circulating EVs were strongly associated with both conventional and thrombogenic risk markers of CVDs, and also with overall CVD risk, highlighting a potentially important role for EVs in CVDs

Chronic lateral ankle instability using anterior tibiofibular ligament distal fascicle transfer augmentation repair: an anatomical, biomechanical, and histological study

Background: The transfer of the anterior tibiofibular ligament distal fascicle (ATiFL-DF) for the augmentation repair of the anterior talofibular ligament (ATFL) shows potential as a surgical technique. However, evidences on the benefits and disadvantages of this method in relation to ankle joint function are lacking.Purpose: This study aimed to provide comprehensive experimental data to validate the feasibility of ATiFL-DF transfer augmentation repair of the ATFL.Methods: This study included 50 embalmed ankle specimens to measure various morphological features, such as length, width, thickness, and angle, for evaluating similarities between the ATiFL-DF and ATFL. Furthermore, 24 fresh-frozen ankle specimens were examined for biomechanical testing of the ATiFL-DF transfer augmented repair of the ATFL. Finally, 12 pairs of ATiFL-DF and ATFL tissues from fresh-frozen ankle specimens were treated with gold chloride staining to analyze mechanoreceptor densities.Results: Anatomical studies found that the lengths and thicknesses of the ATFL and ATiFL-DF are similar. Biomechanical outcomes showed that performing ATiFL-DF transfer for ATFL repair can improve the stability of the talus and ankle joints. This is evident from the results of the anterior drawer, axial load, and ultimate failure load tests. However, performing ATiFL-DF transfer may compromise the stability of the distal tibiofibular joint, based on the Cotton and axial load tests at an external rotation of 5°. Analysis of the histological findings revealed that mechanoreceptor densities for four types of mechanoreceptors were comparable between the ATiFL-DF and ATFL groups.Conclusion: ATiFL-DF transfer is a viable method for augmenting ATFL repair. This technique helps to improve the stability of the talus and ankle joints while compensating for proprioception loss. Although ATiFL-DF transfer augmented repair of the ATFL may negatively affect the stability of the distal tibiofibular joint, this procedure can enhance the stability of the talus and ankle joints

Vascular risk factors for idiopathic normal pressure hydrocephalus: a systematic review and meta-analysis

ObjectiveIdiopathic normal-pressure hydrocephalus (iNPH) is a treatable cause of dementia; however, its etiology and pathogenesis remain poorly understood. The objective of this study was to investigate the prevalence and impact of vascular risk factors in patients with iNPH compared to a control cohort to better understand the potential mechanisms and preventive measures.MethodsWe systematically searched PubMed, Web of Science, Embase, and the Cochrane Library (from inception to December 20, 2022) for studies reporting vascular risk factors for the development of iNPH. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using random-effects models.ResultsAfter screening 1,462 articles, 11 case-control studies comprising 1,048 patients with iNPH and 79,668 cognitively unimpaired controls were included in the meta-analysis. Our data showed that hypertension (N = 991, OR = 2.30, 95% CI 1.64 to 3.23, I2= 64.0%), diabetes mellitus (DM) (N = 985, OR = 3.12, 95% CI 2.29 to 4.27, I2= 44.0%), coronary heart disease (CHD; N = 880, OR = 2.34, 95% CI 1.33 to 4.12, I2= 83.1%), and peripheral vascular disease (N = 172, OR = 2.77, 95% CI 1.50 to 5.13, I2= 0.0%) increased the risk for iNPH, while overweight was a possible factor (N = 225, OR = 2.01, 95% CI 1.34 to 3.04, I2= 0.0%) based on the sensitivity analysis. Smoking and alcohol consumption were not associated with iNPH.ConclusionsOur study suggested that hypertension, DM, CHD, peripheral vascular disease, and overweight were associated with iNPH. These factors might be involved in the pathophysiological mechanisms promoting iNPH. These findings require further investigation in future studies.Systematic review registrationhttps://www.crd.york.ac.uk/PROSPERO/, CRD42022383004

Mode of induction of platelet-derived extracellular vesicles is a critical determinant of their phenotype and function

Platelet-derived extracellular vesicles (PDEVs) are the most abundant amongst all types of EVs in the circulation. However, the mechanisms leading to PDEVs release, their role in coagulation and phenotypic composition are poorly understood. PDEVs from washed platelets were generated using different stimuli and were characterised using nanoparticle tracking analysis. Procoagulant properties were evaluated by fluorescence flow cytometry and calibrated automated thrombography. EVs from plasma were isolated and concentrated using a novel protocol involving a combination of size exclusion chromatography and differential centrifugation, which produces pure and concentrated EVs. Agonist stimulation enhanced PDEV release, but did not alter the average size of EVs compared to those produced by unstimulated platelets. Agonist stimulation led to lower negatively-charged phospholipid externalization in PDEVs, which was reflected in the lower procoagulant activity compared to those generated without agonist stimulation. Circulating EVs did not have externalized negatively-charged phospholipids. None of the 4 types of EVs presented tissue factor. The mechanism by which PDEV formation is induced is a critical determinant of its phenotype and function. Importantly, we have developed methods to obtain clean, concentrated and functional EVs derived from platelet-free plasma and washed platelets, which can be used to provide novel insight into their biological functions

AlphaTracker: a multi-animal tracking and behavioral analysis tool

Computer vision has emerged as a powerful tool to elevate behavioral research. This protocol describes a computer vision machine learning pipeline called AlphaTracker, which has minimal hardware requirements and produces reliable tracking of multiple unmarked animals, as well as behavioral clustering. AlphaTracker pairs a top-down pose-estimation software combined with unsupervised clustering to facilitate behavioral motif discovery that will accelerate behavioral research. All steps of the protocol are provided as open-source software with graphic user interfaces or implementable with command-line prompts. Users with a graphical processing unit (GPU) can model and analyze animal behaviors of interest in less than a day. AlphaTracker greatly facilitates the analysis of the mechanism of individual/social behavior and group dynamics

Effects of fish oil-derived N-3 polyunsaturated fatty acids on the generation and functional activities of extracellular vesicles

Background: Extracellular vesicles (EVs) are submicron membrane-bound vesicles released from almost all cells, which affect many pathophysiological processes involved in cardiovascular diseases (CVDs) and may therefore be considered as a novel marker for CVDs. N-3 polyunsaturated fatty acids (n-3 PUFA) have been suggested to play a role in protecting cardiovascular health. However, there is little information about the effect of n-3 PUFA on circulating EV numbers, composition and functional activity in the context of CVDs. Objective: This project investigated the relationships between circulating EVs, cardiovascular risk markers and CVD risk, as well as the chronic effects of fish oil�derived n-3 PUFA on the numbers, fatty acid composition and procogulatory activity of circulating EVs. Design: Subjects (n=40) aged 40-70yrs with moderate risk of CVDs were recruited into a double-blind, randomised crossover trial of fish oil (1.8g/d n-3 PUFA) or control oil (high-oleic safflower oil) for 12 weeks with a 12-week washout. EVs were analysed by Nanoparticle Tracking Analysis (NTA) and fluorescence flow cytometry (FCM). Total lipid fatty acid composition and procoagulatory activity of circulating EVs were analysed by gas chromatography and a thrombin generation assay, respectively. Results: Circulating EV numbers were positively associated with body mass index (BMI), blood pressure (BP), plasma triacylglycerol (TAG) concentration and overall CVD risk. Furthermore, plasma TAG concentration and diastolic blood pressure (DBP) independently predicted total EV numbers based on a multivariate regression model. Fish oil supplementation significantly decreased numbers of circulating EVs, modified their total lipid fatty acid composition and decreased their thrombogenic potential. Conclusion: Circulating EVs are associated with CVD risk and are particularly strongly related with plasma TAG concentration and DBP. They could therefore potentially serve as a novel biomarker for CVD prediction. Dietary n-3 PUFA modifies the numbers, fatty acid composition and function of circulating EVs in a manner which suggests beneficial effects of n-3 PUFA on CVDs

Dataset supporting paper 'Circulating extracellular vesicles are strongly associated with cardiovascular risk markers'

This dataset includes all reports and summaries of raw data supporting the results presented in the paper �Circulating extracellular vesicles are strongly associated with cardiovascular risk markers'. The objective of this paper is to investigate the relationships between circulating extracellular vesicles (EVs), conventional cardiovascular risk markers and thrombogenic markers in subjects with moderate risk of cardiovascular diseases (CVDs), which is the part aim of a larger study investigating the effect of fish oil-derived n-3 polyunsaturated fatty acids on the generation and functional activities of EVs based on a randomised, double-blind, placebo-controlled crossover intervention. The results and findings involved in this dataset relate only to the baseline data prior to any intervention. A total of 40 subjects (aged 40~70yrs) with moderate risk of CVDs were recruited and involved in the study. Approximately 100ml of blood was collected from individuals after overnight fasting for all analysis, in which 35ml was processed for the analysis of circulating EVs, 9ml was taken for the analysis of blood lipids and glucose levels for the conventional cardiovascular markers evaluation and the rest was operated for the thrombogenic markers evaluation. Data about EVs parameters include: i. the numbers and size of circulating total EVs measured by Nanoparticle Tracking Analysis (NTA); ii. the numbers of EV subpopulations (i.e. phosphatidylserine-positive EVs (PS+EVs), platelet-derived EVs (PDEVs) and endothelial-derived EVs (EDEVs)) measured by flow cytometry (FCM); iii. the procoagulatory activity of circulating EVs (in vitro thrombogenic potential in activating tissue factor-dependent thrombin generation) measured by thrombin generation assay. Data about conventional cardiovascular risk markers include i. body mass index (BMI) measured by Tanita; ii. blood pressure measured by upper arm blood pressure monitor; iii. plasma lipid profile (i.e. triacylglycerol (TAG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and glucose concentrations measured by iLab; iv. 10-year CVD risk detected by QRISK2 scoring system. Data about thrombogenic markers include: i. plasma platelet aggregation measured by 96-well high-throughput aggregometry; ii. plasma thrombin generation measured by thrombin generation assay; iii. plasma clot growth and fibrinolysis measured by thrombodynamics analyser