Cholesterol

Serum analysis used commercial kits according to the manufacturer’s instructions; serum albumin (Alb), bilirubin and alanine transaminase (ALT) (Alpha Laboratories); aspartate aminotransferase (AST) and alkaline phosphatase(ALP) (Randox laboratories); Cholesterol (Wako Chemicals GmbH). All kits were adapted for use on a Cobas Fara centrifugal analyser (Roche Diagnostics).

Insulinemia was measured using the Ultrasentitive ELISA kit (Alpco 80-INSMSU-E01, Salem NH). Hepatic triglycerides (TG) were extracted in methanol/chloroform.^{26 (link)} Plasma and hepatic TG were analyzed with Triglycerides Reagent kit (Randox Laboratories, UK). Cholesterol (Randox Laboratories, UK), nonesterified fatty acids (NEFA) (Thermo Scientific, USA) and leptin (Mouse/Rat Leptin Quantikineࣨ ELISA Kit, R&D Systems) were also measured in plasma samples. Alanine (ALT) and aspartate (AST) aminotransferases plasma levels were determined by biochemical analyses at the Quebec Heart and Lung Institute clinical biochemistry department. To determine cytokine concentrations in liver, proteins from 75 mg of tissue were extracted in PBS 1X buffer (Igepal 1% and proteinase inhibitors 1X). Proteins were measured (Pierce BCA Protein Bioassay kit) and standardized for the multiplex assay, which was performed according to the manufacturer’s instructions (Biorad). Analysis of FA profile in liver was achieved at CHU Sainte-Justine Research Center.^{27 (link)}