Cobas mira plus autoanalyser

Anamnesis and anthropometric data, including sex, age, clinical history and medication, were recorded and included in our database. BMI was calculated from the weight and height measurements (kg/m²). A blood sample was obtained from each patient after overnight fasting. Aliquots were prepared for immediate storage at -80°C in the BioBank at our centre prior to use. Standard biochemical parameters, including lipids, apolipoproteins, blood glucose, hsCRP, and transaminases, were measured using colorimetric, enzymatic and immunoturbidimetric assays (Spinreact, SA, Spain; Horiba, SA, Spain), which were adapted to the Cobas Mira Plus Autoanalyser (Roche Diagnostics, Spain).

After the animals were fasted for 4 h, blood samples were collected in heparin. The samples were analyzed for plasma levels of total cholesterol, high-density lipoprotein cholesterol (HDLc), glutamic oxaloacetic transaminase (GOT), and glutamate–pyruvate transaminase (GPT) (all from Spinreact, SA, Barcelona, Spain) by standardized colorimetric and enzymatic methods adapted to the Cobas Mira Plus Autoanalyser (Roche Diagnostics, Barcelona, Spain). The low-density lipoprotein cholesterol (LDLc) concentration was calculated using the Friedewäld formula, and the concentration of rich triglyceride particles (i.e., VLDLc, VLDL cholesterol) was determined by the formula total cholesterol - (HDLc + LDLc). Plasma levels of leptin, resistin, adiponectin (Milliplex^®, Millipore; Billerica, MA, USA), and FABP4 (Bio Vendor Laboratory Medicine Inc., Brno, Czech Republic) were determined by commercial ELISA kits.

Biochemical parameters, lipids, and apolipoproteins were measured using colorimetric, enzymatic and immunoturbidimetric assays (Spinreact, SA, Spain; Wako Chemicals GmbH, Germany; Polymedco, NY, US; CV < 4 %) that were adapted to a Cobas Mira Plus Autoanalyser (Roche Diagnostics, Spain). The lipid profile was analysed according to Spintrol “H” CAL (Spinreact, SA, Spain) GC–MS reference methods. Spintrol “H” Normal (Spinreact, SA, Spain) was used as a quality control. All samples were subjected to the Liposcale test. This advanced lipoprotein test is based on 2D diffusion-ordered (1)H nuclear magnetic resonance (NMR) spectroscopy. This method adds diffusion coefficients to classical NMR determinations to provide a direct measure of mean particle size and number [23 (link)]. CETP activity was measured using a fluorometric assay (BioVision, CA, USA). Lecithin-cholesterol acyltransferase (LCAT) activity was assessed using a fluorometric assay (Calbiochem, CA, USA). Circulating PCSK9 levels were measured using commercial ELISA kits (R&D Systems, MN, USA).