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Millipore
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Journal: International Journal of Molecular Sciences
Article Title: Characterization of Biomarkers of Thrombo-Inflammation in Patients with First-Diagnosed Atrial Fibrillation
doi: 10.3390/ijms25074109
Figure Lengend Snippet: Pro-inflammatory immune cell function in FDAF—Indications for PAR1 activation via thrombin. ( A ) Increased plasma levels of pro-inflammatory mediators, namely tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6), were seen in patients with FDAF. ( B ) Phenotyping via flow cytometry of peripheral blood mononuclear cells (PBMCs) revealed a higher percentage (% of total PBMCs) of circulating pro-inflammatory cells that possess thrombin-activated (cleaved) PAR1 (TNF-α + or IL-6 + cleaved PAR1 + ) in patients with FDAF. The antibody detects fragments of activated PAR1-cleaved-Ser42 protein. Cytokines TNF-α and IL-6 were visualized via intracellular staining of permeabilized cells. ( C – E ) Evaluation of different immune cell subsets expressing PAR1 via flow cytometry. Antigen-presenting cells (APCs; HLA-DR + ) ( C ), non-T cells (CD3 − ) ( D ), and lymphocytes ( E ). Left panel: percentage of a specified immune cell population expressing PAR1. Right panel: Increased mean fluorescence intensity (MFI) of PAR1. Extracellular (surface) PAR1 expression (=PAR1 that is accessible for activation via FIIa) was detected via staining of unpermeabilized PBMCs with a PAR1 antibody. Patients with FDAF ( n = 80) were compared to controls (patients with chronic cardiovascular diseases but without AF) ( n = 20). Results are expressed as single values, median, and p -values.
Article Snippet: The percentage of
Techniques: Cell Function Assay, Activation Assay, Flow Cytometry, Staining, Expressing, Fluorescence