902 autoanalyser

Insulin concentrations were measured using an Abbott AxSYM system (Abbott Laboratories, Abbott Park, USA) by microparticle enzyme immunoassay with an inter-assay coefficient of variation (CV) of 5.4%. Glucose, triglyceride, total cholesterol, HDL-C, LDL-C, free fatty acid, uric acid, and highly-sensitive CRP concentrations were also measured on a Hitachi 902 autoanalyser (Hitachi High Technologies Corporation) by enzymatic colorimetric assay (Roche) with all CVs lower than 3.2%. Active GLP-1 levels were quantified using ELISA kits (Millipore); this assay had a CV of 7.2%.

Plasma insulin (Crystal Chem Inc), Leptin (Crystal Chem Inc) IL‐1β, IL‐6, and TNFα concentrations (Quantikine kits; R&D Systems Europe, Abingdon, UK) were measured enzymatically. HOMA‐IR was calculated as [fasting glucose × fasting insulin/22.5]. Maternal E18 and weanling P24 plasma samples were thawed and analyzed for concentrations of free fatty acids (FFA), triglycerides (TAG), low‐density lipoprotein cholesterol (LDL), total cholesterol, high‐density lipoprotein low serum‐3 cholesterol (HDLC3), lactate dehydrogenase, lipoprotein lipase, bilirubin, creatinine, uric acid, alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate transaminase (AST), total protein, albumin, and creatine kinase (CK). This analysis was performed using enzymatic colorimetric assays on a Hitachi 902 autoanalyser (Hitachi High Technologies Corporation, Tokyo, Japan).

Blood samples were drawn from an antecubital vein without stasis, after overnight fasting. The samples were centrifuged and the plasma and serum were separated and stored at –70 °C until assayed. Osteoprotegerin (OPG) was determined using ELISA (MicroVue Eia Kit. Quidel Corp. Specialty Products, San Diego, CA, USA). The intra-assay precision was 3% and the inter-assay precision was 4.5%, with a limit of detection of 1.16 to 60 pmol/L. N-MID osteocalcin and intact parathormone (iPTH) were analyzed by electrochemiluminescence immunoassay (Elecsys Modular Analytics 2010 Roche, Mannheim, Germany) The intra and inter coefficients of variation (% CoV) were 2.5% and 2.0%, respectively. Serum phosphorous (P), serum albumin (Alb), and albumin-corrected calcium (cCa) were calculated with the formula: cCa = (Ca (mg/dL) + 0.8(4-Alb g/dL)); total cholesterol (Chol), glucose (Glu), creatinine (Cr), total alkaline phosphatase (tALP), and high-sensitivity C-reactive protein (hsCRP) were measured using standard techniques (Hitachi 902 autoanalyser, Tokyo, Japan). Twenty-four-hour urine and dialysate collection was performed for both CAPD and APD patients.

Plasma samples were analyzed for concentrations of free fatty acids (FFA), triglycerides (TAG), low-density lipoprotein cholesterol (LDL), high-density lipoprotein low serum (HDL). Analysis was performed using enzymatic colorimetric assays using a Hitachi 902 autoanalyser (Hitachi High Technologies Corporation, Tokyo, Japan).