Intradialytic cardiovascular injury is lowest in high-volume haemodiafiltration: a randomized cross-over trial in four intermittent dialysis strategies.

Background: Intradialytic hypotension (IDH) and subsequent tissue damage may contribute to the poor outcome of chronic haemodialysis (HD) patients. While the IDH-incidence is lower in high-volume haemodiafiltration (HV-HDF) than in standard HD (S-HD), survival is better in HV-HDF. Tissue injury, as measured by extracellular vesicle (EV)-release, was compared between four modalities.

Protocol for Measuring Concentrations of Extracellular Vesicles in Human Blood Plasma with Flow Cytometry.

Extracellular vesicles (EVs) are lipid membrane enclosed particles that are released from cells into body fluids, such as blood. EVs offer potential new biomarkers of diseases, because the cellular origin, composition, concentration, and function of EVs change in health and disease. The concentration of EVs from specific cell types in blood can be determined with flow cytometry.

Rate zonal centrifugation can partially separate platelets from platelet-derived vesicles.

Background: Centrifugation is commonly used as a first step to enrich biomarkers from blood. Biomarkers are separated on the basis of density and/or diameter. However, the centrifugation protocol affects the yield and purity of biomarkers, for example, isolation of platelets results in co-isolation with extracellular vesicles (EVs).

Platelet removal by single-step centrifugation.

The study of extracellular vesicles (EVs) in plasma requires removal of cells including platelets. At present, a two-step centrifugation protocol is recommended and commonly used. A simpler protocol that is less operator dependent is likely to improve the quality of plasma samples collected for EV research.

Label-free identification and chemical characterisation of single extracellular vesicles and lipoproteins by synchronous Rayleigh and Raman scattering.

Extracellular vesicles (EVs) present in blood originate from cells of different origins such as red blood cells (RBCs), platelets and leukocytes. In patients with cancer, a small portion of EVs originate from tumour cells and their load is associated with poor clinical outcome. Identification of these tumour-derived extracellular vesicles (tdEVs) is difficult as they are outnumbered by EVs of different tissue of origin as well a large number of lipoproteins (LPs) that are in the same size range.

Extracellular vesicles and coagulation in blood from healthy humans revisited.

: In 2001, we studied the presence and coagulant properties of "microparticles" in the blood of healthy humans. Since then, multiple improvements in detection, isolation and functional characterization of the now called "extracellular vesicles" (EVs) have been made, and shortcomings were identified. : To revisit the presence and function of EVs in blood from healthy humans.

Cellular origin and microRNA profiles of circulating extracellular vesicles in different stages of diabetic nephropathy.

Background: Diabetic nephropathy (DN) is a major complication of diabetes and the main cause of end-stage renal disease. Extracellular vesicles (EVs) are small cell-derived vesicles that can alter disease progression by microRNA (miRNA) transfer.

Methods: In this study, we aimed to characterize the cellular origin and miRNA content of EVs in plasma samples of type 2 diabetes patients at various stages of DN.

The P4-ATPase ATP9A is a novel determinant of exosome release.

Extracellular vesicles (EVs) released by cells have a role in intercellular communication to regulate a wide range of biological processes. Two types of EVs can be recognized. Exosomes, which are released from multi-vesicular bodies upon fusion with the plasma membrane, and ectosomes, which directly bud from the plasma membrane.

Urinary mitochondrial DNA associates with delayed graft function following renal transplantation.

Background: Ischaemia-reperfusion (IR) injury is an important determinant of delayed graft function (DGF) affecting allograft function. Mitochondrial DNA (mtDNA) is released upon cell death and platelet activation into the extracellular environment and has been suggested to be a biomarker in several diseases. Whether extracellular mtDNA accumulates in plasma and/or urine upon renal IR and predisposes DGF is unknown.

Tissue Factor Coagulant Activity is Regulated by the Plasma Membrane Microenvironment.

Background: Tissue factor (TF) can be present in a non-coagulant and coagulant form. Whether the coagulant activity is affected by the plasma membrane microenvironment is unexplored.

Objective: This article studies the presence and coagulant activity of human TF in plasma membrane micro-domains.

Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry.

Background: Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry.

Handling and storage of human body fluids for analysis of extracellular vesicles.

Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze-thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated.

Changes in microparticle numbers and cellular origin during pregnancy and preeclampsia.

Background: Microparticles (MP) are pro-coagulant vesicles derived from various cells. Evidence is accumulating that MP are of pathophysiological relevance in autoimmune, cardiovascular, and thromboembolic diseases and inflammatory disorders. Therefore, their role in the development of preeclampsia was investigated and MP from preeclamptic patients influenced endothelial-dependent vasodilatation.

Simvastatin-induced endothelial cell detachment and microparticle release are prenylation dependent.

Statins reduce cardiovascular disease risk and affect endothelial function by cholesterol-dependent and independent mechanisms. Recently, circulating (detached) endothelial cells and endothelial microparticles (EMP) have been associated with endothelial functioning in vitro and in vivo. We investigated whether simvastatin affects endothelial detachment and release of EMP.

Platelet microparticles contain active caspase 3.

During storage, platelets undergo processes resembling apoptosis, including microparticle release, aminophospholipid exposure, and procaspase 3 processing. Recently, we showed that microparticles from endothelial cells contain caspase 3, one of the executioner enzymes of apoptosis. In this study we determined whether platelet-derived microparticles (PMP) contain caspase 3 in vitro (stored platelet concentrate) and ex vivo (plasma from healthy humans).

Inhibition of microparticle release triggers endothelial cell apoptosis and detachment.

Endothelial cell cultures contain caspase 3-containing microparticles (EMP), which are reported to form during or after cell detachment. We hypothesize that also adherent endothelial cells release EMP, thus protecting these cells from caspase 3 accumulation, detachment and apoptosis. Human umbilical vein endothelial cells (HUVEC) were incubated with and without inhibitors of microparticle release (Y-27632, calpeptin), both in the absence or presence of additional "external stress", i.

Microparticle-associated P-selectin reflects platelet activation in preeclampsia.

Platelet activation in preeclampsia is reflected by elevated levels of platelets exposing P-selectin. In plasma, a non-cell bound (soluble) form of P-selectin is present. Elevated levels of this soluble form have been reported in preeclampsia.

Expression of epidermal growth factors and their receptors in the bronchial epithelium of subjects with chronic obstructive pulmonary disease.

Smoking may affect epithelial repair and differentiation differentially in smokers with and without chronic obstructive pulmonary disease (COPD). We hypothesized that epithelial repair is disturbed in patients with COPD owing to higher expression of epidermal growth factor (EGF)-like factors and/or receptors. We studied epithelial expression of EGF, transforming growth factor a, amphiregulin, heregulin (HRG), betacellulin (BTC), and their receptors, EGFR, HER-2, and HER-3, by immunohistochemical analysis in resected bronchial tissue from 20 subjects with (forced expiratory volume in 1 second [FEV(1)] <75% of predicted value) and 18 without (FEV(1) >85% predicted value) COPD.