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- W2040257440 abstract "Matrix metalloproteinases (MMPs) are calcium/zinc-dependent endoproteinases involved in physiologic and pathologic processes, modulating extracellular matrix degradation (1). MMP-2 (EC 3.4.24.24) and MMP-9 (EC 3.4.24.35) circulating in the peripheral blood (PB) of patients with neoplasia showed contrasting results, revealing the preanalytical issue of whether the method of PB sampling influences MMP concentrations (2) and their zymographic profiles (3). We therefore analyzed the effects of anticoagulants and cell separation media on PB gelatinolytic profiles.PB samples from 30 healthy volunteers were collected into Vacutainer™ Tubes with clot activator (SST), lithium heparin (LH), dipotassium EDTA (K2E), sodium fluoride/potassium oxalate (NaF/KOx), and buffered/acidic citrate [natrium citrate (9NC), acid-citrate-dextrose (ACD), and citrate-phosphate-dextrose-adenine (CPDA); Becton Dickinson]. After centrifugation at 500 g for 15 min at 4 °C, the supernatants and buffy coats were collected and analyzed. Leukocyte subpopulations were obtained after Lympholyte® gradient (5.64% Nycograde™ Polysucrose 400, 9.65% sodium diatrizoate; Cedarlane), and their subset recovery was tested through cytometric analysis (4). Gelatinases from leukocytes and plasma samples were analyzed by gelatin zymography, with 150 μg of total protein loaded on the gel (3). Calibrators were prepared from capillary PB (5). MMP-2 and MMP-9 were measured by ELISA (5). Differences …" @default.
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- W2040257440 date "2003-11-01" @default.
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- W2040257440 title "Effect of Anticoagulants and Cell Separation Media as Preanalytical Determinants on Zymographic Analysis of Plasma Matrix Metalloproteinases" @default.
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- W2040257440 doi "https://doi.org/10.1373/clinchem.2003.022145" @default.
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