Sarah Bos

23 Hemostatic profiles are similar across all etiologies of cirrhosis. 2 manufacturer). Reagents were purchased from Thrombinoscope BV, Maastricht, the Netherlands, and thrombin generation experiments were executed following protocols provided by Thrombinoscope. The following parameters were recorded: endogenous thrombin potential (ETP; which represents the total enzymatic work performed by thrombin during the time that it was active), peak, velocity index (slope between the end of lag time and peak thrombin), and lag time (time needed for thrombin concentration to reach 1/6th of the peak concentration). Conventional coagulation assays The prothrombin time (PT) and activated partial thrombin time (APTT) were assessed on an automated coagulation analyzer (ACL 300 TOP) with reagents (Recombiplastin 2G for PT and Hemosil SynthaSil for APTT) and protocols from the manufacturer (Instrumentation Laboratory, Breda, the Netherlands). Clot lysis time Lysis of a tissue factor-induced clot by exogenous tissue plasminogen activator (tPA) was studied by monitoring changes in turbidity during clot formation and subsequent lysis as described previously.(19) Clot lysis time was determined as the time from the midpoint of the clear to maximum turbid transition, which characterizes clot formation, to the midpoint of the maximum turbid to clear transition, which represents clot lysis. Fibrin structure The average pore size of the fibrin clot was determined in permeation studies as described previously.(20,21) The permeability coefficient Ks was calculated following Darcy’s Law. Plasma clots were generated as described previously.(22) Levels of fibrinogen were assessed on an automated coagulation analyzer (ACL 300 TOP). We used QFA thrombin (Hemosil) and testing was performed according to protocols from the manufacturer (Instrumentation Laboratory, Breda, the Netherlands). Plasma markers of primary hemostasis Soluble P-selectin (sP-selectin) levels were determined using a commercially available enzyme-linked immunosorbent assay (ELISA) (R&D Systems, Abingdon, UK). Levels VWF were assessed with in-house ELISA using commercially available polyclonal antibodies against VWF (DAKO, Glostrup, Denmark). Plasma activity of a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) was measured using the FRETS-VWF73 assay (Peptanova, Sandhausen, Germany) which is described in detail previously.(23) Plasma samples