Cobas c 111 chemistry analyzer

BUN, urine glucose and urine and serum creatinine were determined using Cobas C-111 chemistry analyzer (Roche). β-Hydroxybutyrate was determined using a colorimetric assay (Cayman). Plasma insulin was determined using an ELISA kit (Crystal Chem). Plasma BCAA were determined using a colorimetric kit (BioVision). Urine albumin and KIM-1 were measured by ELISA kits (Bethyl Laboratories and R&D Systems). Urinary sodium, calcium, phosphate, and uric-acid concentrations were determined by colorimetric methods (Lehmann).

Total body fat and lean masses were determined by EchoMRI-100H (Echo Medical Systems LLC, Houston, TX, USA).
HDL and LDL measurements were done using the Cobas C-111 chemistry analyzer (Roche, Switzerland).

Serum and urine levels of creatinine as well as the serum levels of cholesterol, triglycerides (TG), high-density lipoprotein (HDL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and glucose were determined using the Cobas C-111 chemistry analyzer (Roche, Switzerland). Low-density lipoprotein (LDL) levels were calculated by using the following equation: LDL-C = (0.9 × TCHOL) − (0.9 × TG/5) − 28. Creatinine clearance was calculated using the urine and serum creatinine levels (CCr mL/h = Urine creatinine mg/dL × Urine volume/Serum creatinine mg/dL × 24 h). Fasting blood glucose was measured using the Contour^® glucometer (Bayer, Pittsburgh, PA, USA). Serum insulin was determined using an Ultra-Sensitive Mouse Insulin ELISA kit (Crystal Chem, Inc., Elk Grove Village, IL, USA or Millipore, Darmstadt, Germany). Serum leptin was determined by ELISA (Millipore, Darmstadt, Germany).

Serum insulin and urine albumin were measured by ELISAs (Millipore and Bethyl Laboratories, respectively). Serum urea, urine/serum glucose, and creatinine were determined using a Cobas C-111 chemistry analyzer (Roche, Switzerland). Blood urea nitrogen (BUN) was calculated by the serum urea levels (BUN mg/dL = Urea mM × 2.801). CCr was calculated using urine and serum creatinine levels (CCr mL h⁻¹ = Urine creatinine mg/dL × Urine volume/Serum creatinine mg/dL × 24 h). The urine levels of KIM-1 were measured by ELISA kit (R&D Systems, MN, USA).

Plasma metabolites (triglycerides and total protein) were quantified by a photometric method using Cobas™ C111 Chemistry Analyzer (Roche Diagnostics International Ltd. Rotkruez, Switzerland). The system was calibrated using tetramethylammonium chloride (C.f.a.s Calibrator; Roche diagnostics GmbH, Mannheim Germany). Analytic controls (PreciControl ClinChem Multi 1 and 2; Roche diagnostics GmbH, Mannheim Germany) consisting of lyophilized human sera were used for quality control. Triglycerides were quantified using the TRIGL kit (Roche diagnostics GmbH, Mannheim, Germany) and total protein concentration was measured using the TP2 Kit (Roche diagnostics GmbH) according to the manufacturer's protocols. Steroid extraction for cortisol analysis was performed according to Aizen et al. (42 (link)) and cortisol concentrations were measured using a cortisol-specific ELISA according to the protocol published by Yeh et al. (43 (link)).