7600 autoanalyzer

Free fatty acids (FFAs) were measured using an enzymatic assay using the acyl-CoA synthetase-acyl-CoA oxidase method with a Hitachi 7600 Autoanalyzer (Hitachi Ltd., Tokyo, Japan). Triglycerides (TGs) and total cholesterol were measured with a Hitachi 7600 Autoanalyzer (Hitachi Ltd., Tokyo, Japan). High-density lipoprotein (HDL) cholesterol was measured using an enzymatic method. Low-density lipoprotein (LDL) cholesterol was calculated by the Friedewald formula: LDL cholesterol = total cholesterol−[HDL cholesterol  + (TG/5)].

Clinical evaluations were performed according to procedures as previously described9 (link)15 (link). In brief, demographic data, medical history, and health habits were recorded by trained physicians. Standing height and body weight without shoes and with light clothes were measured in standard procedures. Body mass index was calculated as body weight (kg) divided by square of height (meters). Waist circumference was measured at the level of the narrowest point between the iliac crest and the rib cage using a non-stretchable tape. Systolic and diastolic blood pressures were measured using an automated sphygmomanometer, with participants in sitting position.
Overnight fasting blood samples were obtained from each participant, and serum samples were separated for biochemical analysis without freezing. The biochemical values, including liver enzymes, serum lipids, glucose, and uric acid, were measured by a Hitachi 7600 autoanalyzer (Hitachi, Tokyo, Japan) using standard protocols. Serum complement C3 levels were assessed using immune-turbidimetric assay by a Hitachi 7600 autoanalyzer (Hitachi) using standard methods. The coefficients of variation were 4.5% and 6.6% for inter-assay and intra-assay, respectively.

Serum triglyceride and serum total cholesterol concentrations were analyzed by enzymatic assays using a Hitachi 7600 autoanalyzer (Hitachi, Tokyo, Japan). Serum high-density lipoprotein (HDL)-cholesterol concentrations were determined by selective inhibition using a Hitachi 7600 autoanalyzer. Low-density lipoprotein (LDL)-cholesterol concentrations were calculated indirectly using the Friedwald formula; i.e., LDL-cholesterol = total cholesterol − [HDL-cholesterol + (triglyceride/5)] for subjects with serum triglyceride concentrations <400 mg/dL. Serum glucose concentrations were measured according to the hexokinase method on a Hitachi 7600 autoanalyzer.

The lipid profile including total-cholesterol, triglyceride (TG), and free fatty acid (FFA) were measured using a Hitachi 7600 autoanalyzer (Hitachi Ltd., Tokyo, Japan). For separating HDL-cholesterol by dextran-sulfate magnesium precipitation the enzymatic method was used. The LDL-cholesterol concentration was calculated by the Friedewald equation. Lipoprotein-associated phospholipase A₂ (Lp-PLA₂) activity was measured with a high-throughput radiometric activity assay [21 (link)]. The fasting glucose level was analyzed by hexokinase method with a Hitachi 7600 autoanalyzer. Insulin level was measured with immunoradiometric assay, using commercial kit provided by Immuno Nucleo Corporation (Stillwater, MN, USA) and IR was calculated by homeostasis-model assessment (HOMA).

All participants received a comprehensive physical examination, including height, body weight, and blood pressure. BMI was calculated as weight/height² (kg/m²). Blood samples were obtained from all participants in the morning after a 10-h overnight fast to measure the levels of FPG, HbA_1c and 1,5-AG. A 75-g OGTT was administered to each patient to assay the 2hPG. Standard laboratory measurements were performed^{22 (link)}. Serum 1,5-AG levels were measured by an enzymatic method (GlycoMark; GlycoMark Inc., New York, NY, USA) on a 7600 autoanalyzer (Hitachi, Tokyo, Japan) with inter- and intra- assay coefficients of variation (CVs) of <3.5% and <2.5%, respectively.