Cobas c311 analyser

Blood samples were taken after overnight fasting and immediately transported to the central laboratory of the Gynecologic Obstetrical University Hospital in Poznan for analysis. HbA1c level in whole blood was determined using the turbidimetric inhibition immunoassay (TINIA) (Tina-quant Hemoglobin A1c II test in a Cobas c311 analyser (Roche Diagnostics, Basel, Switzerland)).
The total serum cholesterol, HDL cholesterol and triglyceride (TG) levels were determined with Roche Diagnostics reagents (Cholesterol CHOD-PAP, HDL-C plus, and Triglycerides GPO-PAP, respectively) on a Cobas c501 analyser. The following formula was used to calculate the level of LDL cholesterol: LDL cholesterol = total cholesterol − HDL cholesterol − (TG/5).

After an overnight fast, venous blood samples (<20 mL) were collected, centrifuged at 4 °C within 15 min, and stored at −80 °C. Serum IL-18 (Human IL-18 ELISA, MBL, Nagoya, Japan) and serum or plasma concentrations of 10 other biomarkers of inflammation were measured at the Institute for Clinical Diabetology (German Diabetes Center, Düsseldorf, Germany) using highly sensitive ELISAs as previously described (leptin, adiponectin, soluble intercellular adhesion molecule-1 (sICAM-1), omentin, IL-1 receptor antagonist (IL-1RA), chemerin, fetuin-A, IL-6, fibroblast growth factor-21 (FGF21), soluble Eselectin (sE-selectin)) [41 (link)]. Plasma glucose, LDL and HDL cholesterol, as well as uric acid were determined at the clinic lab of the pediatric clinic Dortmund, Germany, and high-sensitivity C-reactive protein (hsCRP) at the lab of Institute for Clinical Diabetology with a Roche/Hitachi Cobas c311 analyser (Roche diagnostics, Mannheim, Germany).
Plasma insulin concentration was measured at the Laboratory for Translational Hormone Analytics of the University of Giessen with an immunoradiometric assay (IRMA; DRG Diagnostics, Marburg, Germany) and the homeostasis model assessment was used to determine insulin resistance (HOMA-IR) [42 (link)]. GFR was estimated using the new creatinine-based equations according to the Chronic Kidney Disease Epidemiology Collaboration [43 (link)].

The EDTA blood samples were analyzed with a Sysmex KX-21 Hematology Analyzer (Kobe, Japan) to determine white blood cell count (WBC), neutrophil (Neu), lymphocyte (Lym), monocyte (Mon), eosinophil (Eos), and basophil (Bas) content. Plasma cholesterol, triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were determined by Cobas C311 Analyser (Roche Diagnostics, Basel, Switzerland). Plasma immunoglobulin A (IgA; Cusabio Biotech Co., Wuhan, China), immunoglobulin M (IgM; Cusabio Biotech Co., Wuhan, China), immunoglobulin G (IgG; Cusabio Biotech Co., Wuhan, China), cortisol (Cusabio Biotech Co., Wuhan, China), and lysozyme (Wuhan Fine Biotech Co., Ltd., Wuhan, China) were determined using commercial ELISA kits.
Plasma total antioxidant capacity (T-AOC; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), total superoxide dismutase (SOD; Beyotime, China), glutathione peroxidase (GSH-Px; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and malondialdehyde (MDA; Beyotime, China) were assessed using commercial kits.

Plasma soluble fms-like tyrosine kinase-1 (sFLT-1) and placental growth factor (PlGF) were measured using commercially available kits (R&D Systems) according to manufacturer's instructions. Albumin and creatinine were measured using commercially available kits (Roche) analysed using a Roche Cobas C311 Analyser.

Levels of serum CRP were determined by immunoturbidometry using the Cobas C311 analyser (Cobas, Roche Diagnostic, Mannheim, Germany). Serum IL-6 and TNF-α were determined using high-sensitivity ProQuantum immunoassays, performed using a StepOne plus real-time PCR analyser (Thermofisher, Loughborough, UK). Each sample was assayed in duplicate. Limit of detection (LOD) for ProQuantum assays were: IL-6 (0.119 pg/mL) and TNF-α (0.012 pg/mL). IL-6 and CRP were detected in all samples (n = 84). TNF-α was

Blood glucose in plasma samples from the cannula draws at breakfast were measured using a Cobas c311 analyser (Roche, Germany). At lunch, capillary blood glucose concentration was measured using a HemoCue (Ängelholm, Sweden) blood glucose analyser. The glycaemic data were compared using measures of incremental area under the blood glucose response curve (iAUC; mmol/L·min) and for glucose, peak height (BGRmax; mmol/L·min).
Blood hormones (insulin, ghrelin, glucagon, and GLP-1) were determined by Multiplex Elisa analysis (Thermo Fisher Scientific, Waltham, MA). The data are expressed as mean concentration over the 150 min period following commencement of the breakfast test.

Metabolic cages were used for 24-hour urine collections at GD6.5 and GD14.5 time points. Urine was analyzed for albumin and creatinine concentration using Roche Cobas C311 Analyser and commercially available rodent kits (Roche).