Biochemical analyses were conducted at the National University Hospital Referral Laboratory, which is accredited by the College of American Pathologists. Plasma glucose concentrations obtained during LMMTT were analyzed using enzymatic methods (ADVIA 2400, Bayer Diagnostics, Tarrytown, NY, USA), and plasma insulin concentrations using a chemiluminescence assay (ADVIA Centaur Analyzer, Siemens Healthcare Diagnostics, Tarrytown, NY, USA).
Advia 2400
Manufactured by Bayer
Sourced in Germany
The ADVIA 2400 is an automated clinical chemistry analyzer designed for high-throughput testing in hospital and reference laboratories. It is capable of performing a wide range of routine and specialized biochemical assays.
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Lab products found in correlation
9 protocols using advia 2400
1
Biochemical Analyses of Plasma Glucose and Insulin
2
Diabetes Exclusion Criteria in Research
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We excluded individuals with self-reported physician-diagnosed diabetes, use of diabetes medication, or undiagnosed diabetes (fasting glucose (FG) ≥126 mg/dL (7 mmol/L) or random glucose (RG) ≥200 mg/dL (11.1 mmol/L) or HbA1c ≥6.5% (48 mmol/mol) where available). FG was measured after an overnight 8–12 hours’ fast using enzymatic methods (ADVIA 2400; Bayer Diagnostics and Siemens Healthcare Diagnostics). HbA1c was reported in National Glycohemoglobin Standardization Program (NGSP) percent.
3
Cytokine, Procalcitonin, and CRP Measurement
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Samples for cytokine, procalcitonin and C-reactive protein (CRP) determinations were obtained on the first day and after 72h and 7 days of treatment, centrifuged and frozen at -80°C until analysis. Determination of interleukin (IL)- 6, IL-8 and IL-10 levels was performed using a commercial enzyme immunoassay technique (Biosource, Nivelles, Belgium). An immunoluminometric technique was used to measure procalcitonin (Liaison Brahms PCT; DiaSorin, Saluggia, Italy) with a detection limit of 0.3 ng/ml. CRP was measured with an immunoturbidimetric method using a commercially available test (Bayer Diagnostics, Leverkusen, Germany) with an Advia 2400.
4
Evaluating Nitrite/Nitrate and Oxidative Stress Markers
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At baseline, a blood sample and a spontaneous spot urine sample were collected by general practitioners (GPs) during the health checkup and then shipped to the study center and maintained at -80°C until further processing. Urinary concentrations of nitrite/nitrate were determined using the nitrite/nitrate Colorimetric Assay Kit of Cayman Chemical (Ann Arbor, Michigan, USA). This method detects the sum of nitrate and nitrite. Urine samples were used directly after dilution to a proper concentration (1 : 5, 1 : 25, or 1 : 50 depending on the levels of nitrite/nitrate in the urine sample). For renal function adjustment of the spot urine samples, urinary creatinine was determined by the kinetic Jaffe method. In addition, the acute-phase inflammatory protein C-reactive protein (CRP) was measured in serum samples by immunoturbidimetry with the wrCRP antibody (Bayer, Leverkusen, Germany) on the ADVIA 2400. Furthermore, the levels of an established oxidative stress marker, urinary 8-isoprostane, were determined by the 8iso1 ELISA kit from Detroit R&D (Detroit, Michigan, USA).
5
Liver Biochemical Tests Protocol
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Liver biochemical tests (AST, ALT, and ALP) were performed on a clinical chemistry automatic analyzer (ADVIA 2400, Bayer Diagnostics). AST, ALT, and ALP were measured according to the previous methods [20 (link), 21 ] using commercial assay kit (Bayer Diagnostics). The serum total concentrations of bilirubin were measured adhering to the scientific methods [22 (link), 23 (link)] utilizing commercial kits on the Express Plus biochemical analyzer (Ciba-Corning Diagnostics).
6
Serum Lipid Profile Measurement
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Blood (9 mL) was drawn from the central ear artery in the morning after a 14-h fast before the beginning of the experiment (W0), at the end of the fourth, eighth, and twelfth week of the experiment (W4, W8, W12, respectively), and it was centrifuged (1500 × g for 15 min at 4°C) to separate the serum. Then, the serum samples were stored at –80°C until analysis. Serum concentrations of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglyceride were measured by the enzymatic method using an automatic biochemical analyzer (ADVIA-2400, Bayer, Germany) [Figure 1 B].
7
Standardized Fasting Lipid Profiling
8
Blood Lipid Profile Monitoring
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At the beginning of the experiment (week 0) and the end of the 6th (week 6) and 12th weeks (week 12), blood was collected from the central ear artery the morning following a 14-h fast. Serum samples were separated by centrifugation at 4 °C, 1500 × g for 15 min and stored at -80 °C until analysis. Serum concentrations of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were determined using an autoanalyzer (ADVIA-2400, Bayer, Germany) [17 (link)].
9
Lipid and Glucose Profiling in Plasma
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After 12-h fasting before sacrifice at various ages, four groups (n = 10 in each group per age) of peripheral blood were obtained. Plasma concentrations of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and glucose (GLU) were measured using COD-PAP and GPO-PAP methods with automatic analyzer BAYER ADVIA-2400 (Germany), as described in Ref. [19 (link)]. The plasma insulin concentration was determined using ultrasensitive ELISA kit (Kongcheng, Shanghai, China).
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