490 aggregometer

Platelet aggregation was measured in PRP, using the Chrono-Log 490 aggregometer (CHRONO-LOG, Havertown, PA, USA). PRP samples were pre-incubated with the examined extract/standards at the final concentrations of 1–50 µg/mL for 15 min at 37 °C and transferred into aggregometer cuvettes. Aggregation was induced by ADP (at the final concentration of 10 µM) or collagen (at the final concentration of 2 µg/mL). Control samples were untreated with the examined analytes.
TT, PT, and aPTT were measured in fresh blood plasma with the use of Kselmed K-3002 Optic coagulometer (Grudziądz, Poland), using reagents purchased from Diagon Kft. (Budapest, Hungary). Plasma samples were pre-incubated with the examined analytes at the final concentration of 1–50 µg/mL for 15 min at 37 °C. Control samples were untreated with the examined analytes. Argatroban was used as a positive control.

Platelet aggregation in PRP or WP was done under stirring conditions (1000 rpm) at 37°C using light transmission aggregometry with a CHRONO-LOG 490 aggregometer (CHRONO-LOG, Havertown, PA, USA). WP reactions were supplemented with 1 mg/ml human fibrinogen and 0.2 mg/ml human IgGs before experimentation. Platelets were stimulated with bacteria and other agonists, including crosslinked collagen-related peptide (CRP-xl), thrombin receptor activator peptide 6 amide (TRAP-6) and ADP. To crosslink FcγRIIA, platelets were incubated with anti-FcγRIIA monoclonal antibody (mAb) (4 μg/ml, clone IV.3) for 2 minutes, followed by the addition of crosslinking F(ab’)₂ rabbit anti-mouse IgG (30 μg/ml), altogether designated as IV.3-xl hereafter.

Platelet aggregation was measured in PRP, using the Chrono-Log 490 aggregometer (CHRONO-LOG, Havertown, PA, USA). PRP samples were pre-incubated with the examined extract/standards at the final concentrations of 1–50 µg/mL for 15 min at 37 °C and transferred into aggregometer cuvettes. Aggregation was induced by ADP (at the final concentration of 10 µM) or collagen (at the final concentration of 2 µg/mL). Control samples were untreated with the examined analytes. Aspirin DL-lysine was used as positive control.
Thrombin (TT), prothrombin (PT) and the activated partial thromboplastin time (aPTT) were measured in fresh blood plasma with the use of a Kselmed K-3002 Optic coagulometer (Kselmed, Grudziądz, Poland), using reagents purchased from Diagon Kft. (Budapest, Hungary). Plasma samples were pre-incubated with the examined extract/standards at the final concentration of 1–50 µg/mL for 15 min at 37 °C. Control samples were untreated with the examined analytes. Argatroban was used as positive control.