Sarah Bos

40 CHAPTER 3 drugs between healthy lean controls, healthy obese controls, patients with NASH- related cirrhosis, and patients with cirrhosis related to alcohol abuse. Methods Patients Twenty-two patients with known and well-defined NASH-related cirrhosis were included. These patients were under routine control at the department of internal medicineof theVirginiaCommonwealthUniversity inRichmond, USA. Thediagnosis of NAFLD-related cirrhosis was made through liver biopsy, which was graded according to the NASH Clinical Research Network (NASH CRN) scoring system, and the NAFLD activity score (NAS) was based on the unweighted sum of steatosis, lobular inflammation, and hepatocellular ballooning scores.(11) Exclusion criteria consisted of a documented history of congenital coagulation disorders, presence of active infection ( < 2 weeks), use of anticoagulant drugs in the past 10 days, pregnancy, human immunodeficiency virus (HIV) positivity and recent (7 days) transfusion with blood products. The results of the patients with NASH-related cirrhosis were compared to the results of fifteen patients with cirrhosis related to alcohol abuse (alcoholic steatohepatitis [ASH]-related cirrhosis), and to two control groups, which consist of lean (BMI < 25kg/m2; n=20) and overweight (BMI > 25kg/m2; n=20) subjects with no evidence of liver disease or hepatic steatosis. The local medical ethical committee approved this study protocol and written informed consent was obtained from each subject before inclusion. Thrombin generation testing Potency of three anticoagulants was tested in patients and controls by thrombin generation testing. The thrombin generation test was performed using platelet- poor plasma (PPP) with the fluorimetric method.(12) To activate coagulation, a commercially available reagent containing recombinant tissue factor (TF), phospholipids and soluble thrombomodulin was used (Thrombinoscope BV, Maastricht, The Netherlands). All procedures were according to the protocol suggested by Thrombinoscope B.V. The potency of the anticoagulant drugs was expressed as the percentual change in various parameters of the thrombin generation curve (endogenous thrombin potential (ETP), peak, velocity index, and lag time) after the addition of anticoagulants. These percentages were compared between groups.