Sarah Bos

93 Efficacy of pro- and anticoagulant strategies in plasma 6 mentionedagents.Coagulationwasactivatedusingcommerciallyavailablereagents containing recombinant tissue factor (final concentration 5 pM), phospholipids (final concentration 4 μ M), in the presence of soluble thrombomodulin (TM, the concentration of which is not revealed by the manufacturer). These reagents were purchased from Thrombinoscope BV, Maastricht, The Netherlands. Thrombin Calibrator (Thrombinoscope BV) was added to calibrate the thrombin generation curves. For each plasma sample, we used a single calibration using plasma that was not spiked with pro- or anticoagulants. This calibrator was also used for the samples to which the various pro- and anticoagulants were added. A fluorogenic substrate with CaCl2 (FluCa-kit, Thrombinoscope BV, Maastricht, The Netherlands) was dispensed in each well to allow a continuous registration of thrombin generation. Fluorescence was read in time by a fluorometer, Fluoroskan Ascent® (ThermoFisher Scientific, Helsinki, Finland). All procedures were undertaken according to the protocol suggested by Thrombinoscope B.V. The pro- or anticoagulant potency of the different agents was expressed as the percentual change of endogenous thrombin potential (ETP) after addition of the study agent. We calculated the percentage of change in ETP for each individual sample, and compared the median change in ETP between patients and controls. Coagulation parameters. The INR was assessed with commercially available methods on an automated coagulation analyser (ACL 300 TOP) with reagents (Recombiplastin 2G) and protocols from the manufacturer (Instrumentation Laboratory, Breda, the Netherlands). Levels of fibrinogen (Fg) and antithrombin were assessed on an automated coagulation analyser (ACL 300 TOP). We used Q.F.A. Thrombin (Hemosil) for fibrinogen and Liquid Antithrombin for antithrombin. Testing was performed according to the protocols from the manufacturer (Instrumentation Laboratory). Statistical analyses Data are expressed as means (with standard deviations [SDs]), medians [with interquartile ranges], or numbers (with percentages) as appropriate. Multiple groups were compared using One-way ANOVA or Kruskal-Wallis H test as appropriate. P values of 0.05 or less were considered statistically significant. Statistical analyses were performed with Graph Pad Prism (San Diego, USA) and IBM SPSS Statistics 23.0 (IBM, Chicago, USA).