LitCovid-sentences  

PMC:7172841 / 30802-34052 JSONTXT 11 Projects

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Id Subject Object Predicate Lexical cue
T245 0-14 Sentence denotes Method Details
T246 15-268 Sentence denotes White blood cells were incubated for 15 min in the dark with the monoclonal antibodies anti-CD14 FITC, anti-HLA-DR-PE, anti-CD45 PC5 (Beckman Coulter, Marseille, France) and Quantibrite HLA-DR/anti-monocyte PerCP-Cy5 (Becton Dickinson, Cockeysville Md).
T247 269-596 Sentence denotes White blood cells were also incubated for 15 min with anti-CD3 FITC, anti-CD4 FITC and anti-CD19 FITC (fluorescein isothiocyanate, emission 525 nm, Beckman Coulter); with anti-CD4 PE, anti-CD8 PE, and anti-CD(16+56) PE (phycoerythrin, emission 575nm, Beckman Coulter); and with anti-CD45 PC5 (emission 667 nm, Beckman Coulter).
T248 597-680 Sentence denotes Fluorospheres (Beckman Coulter) were used for the determination of absolute counts.
T249 681-794 Sentence denotes Cells were analyzed after running through the CYTOMICS FC500 flow cytometer (Beckman Coulter Co, Miami, Florida).
T250 795-872 Sentence denotes Isotypic IgG controls stained also with anti-CD45 were used for each patient.
T251 873-975 Sentence denotes Results were expressed as absolute molecules of HLA-DR on CD14/CD14+CD45+ cells, percentages, and MFI.
T252 976-1283 Sentence denotes In separate experiments, whole blood was treated with 10 μg/mL brefeldin A (Invitrogen, Carlsbad, CA, USA) for 60 min at 37°C in 5% CO2 followed by incubation in the dark for 15 min at room temperature with anti-CD4 FITC, anti-CD14 FITC (Beckman Coulter) and anti-CD45 PC5 followed by red blood cells lysis.
T253 1284-1441 Sentence denotes Stained cells were then washed with Dulbecco’s Phosphate Buffered Saline and permeabilized with the IntraPrep Parmeabilization Reagent kit (Beckman Coulter).
T254 1442-1517 Sentence denotes Anti-IL-6 PE (Beckman Coulter) was added and analyzed after flow cytometry.
T255 1518-1667 Sentence denotes Peripheral blood mononuclear cells (PBMCs) were isolated after gradient centrifugation over Ficoll (Biochrom, Berlin, Germany) for 20 min at 1,400 g.
T256 1668-1793 Sentence denotes After three washings in ice-cold PBS pH 7.2, PBMCs were counted in a Neubauer plate with trypan blue exclusion of dead cells.
T257 1794-2010 Sentence denotes They were then diluted in RPMI 1640 enriched with 2mM of L-glutamine, 500 μg/mL of gentamicin, 100 U/mL of penicillin G, 10 mM of pyruvate, 10% fetal bovine serum (Biochrom) and suspended in wells of a 96-well plate.
T258 2011-2083 Sentence denotes The final volume per well was 200 μL with a density of 2 × 106 cells/mL.
T259 2084-2174 Sentence denotes PBMCs were exposed in duplicate for 24 h or 5 days at 37°C in 5% CO2 to different stimuli:
T260 2175-2396 Sentence denotes 10 ng/mL of Escherichia coli O55:B5 lipopolysaccharide (LPS, Sigma, St. Louis, USA); 5 × 105 colony forming units of heat-killed Candida albicans; 25% of patient plasma; or 10 μg/mL of Tocilizumab (Roche, Athens, Greece).
T261 2397-2466 Sentence denotes After incubation, cells were removed and analyzed for flow cytometry.
T262 2467-2646 Sentence denotes Concentrations of TNFα, IL-1β, IL-6, IL-17A and IFNγ were measured in cell supernatants and/or serum in duplicate by an enzyme immunoassay (Invitrogen, Carlsbad, California, USA).
T263 2647-2803 Sentence denotes The lowest detections limits were as follows: for TNF-α, 40 pg/mL; for IL-1β, 10 pg/mL; for IL-6, 10 pg/mL; for IL-17A, 10 pg/mL; and for IFN-γ, 12.5 pg/mL.
T264 2804-2966 Sentence denotes Concentrations of ferritin were measured in serum by an enzyme-immunoassay (ORGENTEC Diagnostika GmbH, Mainz, Germany); the lower limit of detection was 75 ng/mL.
T265 2967-3164 Sentence denotes Six patients with immune dysregulation were treated with single intravenous infusion of 8 mg/kg of Tocilizumab (RoAcremra, Roche) based on the National Health Care off-label provision for COVID-19.
T266 3165-3250 Sentence denotes Absolute lymphocyte blood counts were compared before and after Tocilizumab infusion.