Using enzyme-linked immunosorbent assays, we measured serum hyaluronan (R&D Systems, Minneapolis, MN) and syndecan-1 (Diaclone, Besancon, France) as markers of the glycocalyx, and serum von Willebrand Factor (vWF), (Assaypro, Cambridge, MA) and vascular cell adhesion molecule (VCAM-1), ( R&D Systems, Minneapolis, MN) as markers of endothelial dysfunction. All measures were performed in duplicate.
Syndecan 1
Manufactured by Diaclone
Sourced in United States
Syndecan-1 is a cell surface proteoglycan that functions as a receptor for extracellular matrix proteins. It is involved in cell-cell and cell-matrix interactions, signal transduction, and the regulation of cell growth and differentiation.
Automatically generated - may contain errors
4 protocols using syndecan 1
1
Glycocalyx and Endothelial Dysfunction Markers
2
Glycocalyx Imaging and Biomarkers in Chronic Kidney Disease
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Blood was taken for full blood count, urea and electrolytes, and liver function tests. Additional serum and plasma samples were stored at -80°C for subsequent analyses by enzyme-linked immunosorbent assays for markers of the glycocalyx such as hyaluronan (R&D Systems, Minneapolis, MN, USA) and syndecan-1 (Diaclone, Besançon, France), endothelial dysfunction such as vWF (Assaypro, Massachussets) and VCAM-1 (R&D Systems, Minneapolis, MN, USA), and markers of uraemic toxins including PCS and IS by high-performance liquid chromatography.
Glycocalyx imaging was performed using GlycoCheck TM (Maastricht, Netherlands), consisting of a handheld Capiscope camera placed sublingually for automated video capture of the vasculature. The images were then analyzed by the GlycoCheck TM software according to the algorithm described by Lee et al. [15] . In brief, this identifies all available microvessels between 5 and 25 µm in the field of view and detects erythrocytes in blood vessels through light-emitting diodes. By measuring the width of erythrocyte columns and the distance from the erythrocyte-impermeable glycocalyx margin, an averaged perfused boundary region (PBR) reading is calculated [8] . All readings were performed by a single operator (H.L.) to reduce bias.
Glycocalyx imaging was performed using GlycoCheck TM (Maastricht, Netherlands), consisting of a handheld Capiscope camera placed sublingually for automated video capture of the vasculature. The images were then analyzed by the GlycoCheck TM software according to the algorithm described by Lee et al. [15] . In brief, this identifies all available microvessels between 5 and 25 µm in the field of view and detects erythrocytes in blood vessels through light-emitting diodes. By measuring the width of erythrocyte columns and the distance from the erythrocyte-impermeable glycocalyx margin, an averaged perfused boundary region (PBR) reading is calculated [8] . All readings were performed by a single operator (H.L.) to reduce bias.
3
Plasma Biomarker Quantification Protocol
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Peripheral venous blood samples were collected using ethylenediaminetetraacetic acid as an anticoagulant. Samples for enzyme-linked immunosorbent assay (ELISA) were centrifuged (882 g, 7 min, room temperature), the supernatant plasma was then aliquoted and stored at −80°C until further analysis. ELISAs in duplicate were performed in accordance with the manufacturer's instructions to determine plasma concentrations of heparan sulphate (Cusabio Technology, Houston, TX, USA), hyaluronan (Echelon Biosciences, Salt Lake City, UT, USA) and syndecan-1 (Diaclone SAS, Besançon, France).
4
Comprehensive Proteomic Profiling of COVID-19, Sepsis
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The Olink (Uppsala, Sweden) “inflammation1” and “cardiovascular2” proteomic panels comprise 92 proteins each. Seven proteins are common to both panels. A total of 184 proteins in 75 samples (COVID-19, bacterial sepsis, healthy) were measured in one batch to avoid technical variation. Briefly, the Olink proximity extension assay uses two specific oligonucleotide-labeled antibodies per protein (“probes”). When the two probes are in proximity a new PCR target sequence is formed via a proximity-dependent DNA polymerization event. The resulting sequence is subsequently detected and quantified using standard real-time quantitative PCR, as previously reported [19 (link)]. Measurements were conducted in triplicate. Results are reported as arbitrary units on a log2 scale. The proteins included in each panel, measurement details, and validation data are available online (www.olink.com/downloads ).
Plasma levels of the glycocalyx-core protein syndecan-1 (Diaclone, Besançon, France) and the endothelial-specific proinflammatory mediator Angpt-2 (R&D Systems, Minneapolis, USA) were measured using commercially available enzyme-linked immunosorbent assay kits in accordance with the manufacturer’s instructions.
Plasma levels of the glycocalyx-core protein syndecan-1 (Diaclone, Besançon, France) and the endothelial-specific proinflammatory mediator Angpt-2 (R&D Systems, Minneapolis, USA) were measured using commercially available enzyme-linked immunosorbent assay kits in accordance with the manufacturer’s instructions.
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