Inclusion criteria were age >18 years and biopsy-confirmed NAFLD or healthy liver. Exclusion criteria were: liver disease other Selleckchem GSK-3 inhibitor than NAFLD, anticipated

need for liver transplantation within a year, or complications of endstage liver disease such as variceal bleeding or ascites; concurrent medical illnesses, contraindications for liver biopsy; use of medications known to cause or exacerbate steatohepatitis (such as corticosteroids) or antibiotics, pre- or probiotics in the preceding 6 months; consumption of more than 20 g of alcohol/day; use of vitamin E or fish oil supplements; chronic gastrointestinal diseases, previous gastrointestinal surgery modifying the anatomy; pregnancy or lactating state. Patients provided information regarding medication use, alcohol consumption, and smoking history. Past medical and surgical history was recorded and, in addition, data on ethnicity were collected. Height and weight were measured and BMI was calculated. Subjects were asked

to complete the 7-day food records the week prior to liver biopsy (or liver donation). The stool sample was collected at the end of this week and within 24 hours preceding the biopsy. Portion sizes were estimated using the 2D Food Portion Visual chart Protein Tyrosine Kinase inhibitor (Nutrition Consulting Enterprises, Framingham, MA). Food records were analyzed for macro- and micronutrient content using Diet Analysis Plus v. 7.0.1 (Thomson Wadsworth, Stamford, CT). The participants also recorded their physical activities for 7 days during the week preceding the biopsy. They listed the type of activity, duration, and level

of difficulty (mild, moderate, strenuous, very strenuous). Units of exercise were used to estimate physical activity as follows: 1 unit = 30 minutes mild, 20 minutes moderate, 10 minutes strenuous, or 5 minutes very strenuous activity.30 Basal metabolic rate (BMR) was calculated with the Harris-Benedict equation [men: BMR = 66.5 + (13.75 × weight in kg) + (5.003 × height in cm) − (6.755 × age in years); women BMR = 655.1 + (9.563 × weight in kg) + (1.850 × height in cm) − (4.676 × age in years)] and the estimated energy expenditure (EER) was calculated according to Health Canada Guidelines (http://www.hc-sc.gc.ca/fn-an/nutrition/reference/index-eng.php). Fasting plasma glucose Acetophenone was measured by the enzymatic hexokinase method on an Architect c8000 System (Abbot Laboratories, Abbot Park, IL). Serum insulin was determined by radioimmunoassay (Immulite 2500, Siemens Diagnostics, Los Angeles, CA). IR was calculated using the Homeostasis Model Assessment (HOMA)-IR. Hemoglobin A1c in plasma was measured by ion exchange HPLC (Variant II analyzer, Bio-Rad Laboratories, Montreal, QC, Canada). ALT, aspartate aminotransferase (AST), and alkaline phosphatase (ALP) in plasma as well as serum triglycerides and total cholesterol were measured using the Architect c8000 system (Abbot Laboratories).