Five-milliliter blood samples were taken from the forearm venous blood vessels of each subject and placed in Eppendorf tubes. The tubes were centrifuged at 1,500 r/min for 10 min to obtain serum samples for the measurements of biochemical parameters. After labeling in the biochemistry laboratory, all serum samples were stored at –80°C until the day of analysis. Serum urea, creatinine, fasting blood glucose (FBG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), fT3, fT4, C-reactive protein (CRP), glycated hemoglobin (HbA1c), triglycerides (TG), total cholesterol (TC), and high-density and low-density lipoprotein cholesterol (HDL-C and LDL-C) levels were determined using commercially available assay kits (Roche, USA) with an auto-analyzer (Roche Cobas Integra 800 auto-analyzer).
Cobas integra 800 autoanalyzer
Manufactured by Roche
Sourced in Germany, United States, Switzerland
The Cobas Integra 800 is an autoanalyzer instrument designed for clinical chemistry analysis. It is capable of performing a variety of in vitro diagnostic tests on biological samples such as serum, plasma, and urine. The Cobas Integra 800 is equipped with automated sample handling, reagent delivery, and result reporting functionalities to streamline the analytical process.
Automatically generated - may contain errors
Lab products found in correlation
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Mini vidas analyzer, by bioMérieux (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
Ultraspec 3000, by GE Healthcare (1 mentions)
Elecsys autoanalyzer, by Roche (1 mentions)
Hdl c plus kit, by Roche (1 mentions)
7600 autoanalyzer, by Hitachi (1 mentions)
Ficoll paque plus medium, by GE Healthcare (1 mentions)
11 protocols using cobas integra 800 autoanalyzer
1
Comprehensive Blood Panel Analysis
2
Aortic Blood Analysis Protocol
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After the procedure, biochemical analyses were performed on blood samples taken from the abdominal aorta. BUN, creatinine, and amylase tests were performed using COBAS Integra 800 autoanalyzer (Roche Diagnostics GmbH, D-68298 Mannheim, Germany); and troponin I tests were performed using Mini Vidas analyzer (Biomerieux). Analyses for BUN were performed via the kinetic method using urease and glutamate dehydrogenase; those for amylase were performed via the calorimetric enzymatic method, and those for troponin I were conducted via the enzyme-linked fluorescent immunoassay method.
3
Metabolic Biomarkers in Fasting Individuals
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Blood samples were drawn under fasting conditions. Blood levels of UA, glucose, total cholesterol, high density (HDL), and low-density (LDL) lipoprotein cholesterol and triglycerides were measured by enzymatic methods (Roche Diagnostics, Mannheim, Germany). Serum insulin levels were measured using a Cobas Integra 800 Autoanalyzer (Roche Diagnostics, Indianapolis, IN, USA). Ultrasensitive C-reactive protein was measured by CRP (latex) HS Roche kit. Hyperuricemia was defined as above 7 mg/dL in men and above 5.7 mg/dL in women. Insulin resistance was calculated by the homeostasis model of assessment-insulin resistance (HOMA-IR) approach, calculated as insulin (microunits per milliliter) x blood glucose (millimoles per liter)/22.5 [28 (link)].
4
Plasma Glucose Measurement in Ketamine-Anesthetized Rats
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On the 7th or 21st days of the experiments and after a
12-hour fast, we aesthetized the rats with injections
of ketamine [100 mg/kg/ intramuscular (i.m.)].
Intracardiac blood samples were obtained from all
rats and placed in EDTA tubes, after which the rats
were sacrificied. Blood samples were centrifuged
immediately at 1500 xg (10 minutes, 4˚C) to remove
the plasma. Plasma glucose measurements were
performed on a Cobas Integra 800 autoanalyzer
(Roche Diagnostics, Mannheim, Germany). For
the spectrophotometrical measurements, we used
an Ultraspec 3000 (Pharmacia Biotech, Biochrom
Ltd., Cambridge, UK) and for ELISA, an ELx800
(BioTek Instruments, Inc., Winooski, Vermont,
USA).
12-hour fast, we aesthetized the rats with injections
of ketamine [100 mg/kg/ intramuscular (i.m.)].
Intracardiac blood samples were obtained from all
rats and placed in EDTA tubes, after which the rats
were sacrificied. Blood samples were centrifuged
immediately at 1500 xg (10 minutes, 4˚C) to remove
the plasma. Plasma glucose measurements were
performed on a Cobas Integra 800 autoanalyzer
(Roche Diagnostics, Mannheim, Germany). For
the spectrophotometrical measurements, we used
an Ultraspec 3000 (Pharmacia Biotech, Biochrom
Ltd., Cambridge, UK) and for ELISA, an ELx800
(BioTek Instruments, Inc., Winooski, Vermont,
USA).
5
Inhibitory Control and Oxidative Stress
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Inhibitory control test is a computerized test of response inhibition and working memory that is freely downloadable at www.hecme.tv [14 (link)]. While the original version of the test used the English letters X/Y to define targets of the test, we adapted a special modification to the test by using the numbers 7/8 in Arabic (ν/∧) as targets; this modification was created by expert staff members at the Faculty of Computers and Information, Menoufia University, to suit Egyptian patients.
The plasma ammonia test procedure was done using a Cobas Integra 800 Auto analyzer (Roche Diagnostics Ltd., Germany, catalogue number: M, 87432).
Serum 3-nitrotyrosine levels: it was measured using the quantitative sandwich enzyme immunoassay technique (ELISA). Adding the substrate to a complex of 3-nitrotyrosine antigen, antibody, and a catalyzing enzyme; makes it blue. This is followed by adding the HRP catalyzing enzyme. The reaction is terminated by the addition of a sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of 3-nitrotyrosine in the samples is then determined by comparing the O.D. of the samples to the standard curve.
The plasma ammonia test procedure was done using a Cobas Integra 800 Auto analyzer (Roche Diagnostics Ltd., Germany, catalogue number: M, 87432).
Serum 3-nitrotyrosine levels: it was measured using the quantitative sandwich enzyme immunoassay technique (ELISA). Adding the substrate to a complex of 3-nitrotyrosine antigen, antibody, and a catalyzing enzyme; makes it blue. This is followed by adding the HRP catalyzing enzyme. The reaction is terminated by the addition of a sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of 3-nitrotyrosine in the samples is then determined by comparing the O.D. of the samples to the standard curve.
6
Measurement of Renin, Liver, and Kidney Biomarkers
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Renin test procedure was done using ELISA [DRG® Renin ELISA (EIA-5125) Germany] on ELISA reader Multiskan FC (Cat. no. 51119000), Thermo Fisher Scientific (Finland). The procedures were done according to the manufacturer's instructions.
Liver function tests included alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, albumin and kidney functions including urea and creatinine, were measured using Cobas Integra 800 Auto analyzer (Roche Diagnostics Ltd - Germany. Catalogue number; M, 87432). Prothrombin test was done using BFT* II Analyzer (Dade Behring Marburg GmbH, D-35041 Marburg, Germany).
Liver function tests included alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, albumin and kidney functions including urea and creatinine, were measured using Cobas Integra 800 Auto analyzer (Roche Diagnostics Ltd - Germany. Catalogue number; M, 87432). Prothrombin test was done using BFT* II Analyzer (Dade Behring Marburg GmbH, D-35041 Marburg, Germany).
7
Thyroid and Metabolic Biomarker Analyses
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Serum fT3 and fT4 were measured on the Elecsys
autoanalyzer (Roche Diagnostics, Germany). Serum glucose,
total cholesterol (TC), triglyceride (TG) and albumin
levels, alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) activities were assayed on Cobas
Integra 800 autoanalyzer (Roche Diagnostics, Germany).
autoanalyzer (Roche Diagnostics, Germany). Serum glucose,
total cholesterol (TC), triglyceride (TG) and albumin
levels, alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) activities were assayed on Cobas
Integra 800 autoanalyzer (Roche Diagnostics, Germany).
8
Measuring Lipid Profile and Apolipoproteins
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Fasting total cholesterol (TC) and triglyceride (TG) were measured through enzymatic assays using CHOL and TG Kits (Roche, Mannheim, Germany), respectively. Analysis of high-density lipoprotein (HDL-) cholesterol was conducted via selective inhibition using an HDL-C Plus Kit (Roche, Mannheim, Germany). The result of the color reaction was measured with a Hitachi 7600 autoanalyzer (Hitachi, Tokyo, Japan). Low-density lipoprotein (LDL-) cholesterol levels were calculated by the Friedewald formula: LDL-cholesterol = TC – [HDL-cholesterol + (TG/5)].
Serum fasting apolipoprotein B and apolipoprotein A-I were measured via immunoturbidimetric assays with Apolipoprotein B and Apolipoprotein A-I Kits (Roche, Mannheim, Germany), respectively. Turbidity was assessed using a Cobas Integra 800 autoanalyzer (Roche, Rotkreuz, Switzerland).
Serum fasting apolipoprotein B and apolipoprotein A-I were measured via immunoturbidimetric assays with Apolipoprotein B and Apolipoprotein A-I Kits (Roche, Mannheim, Germany), respectively. Turbidity was assessed using a Cobas Integra 800 autoanalyzer (Roche, Rotkreuz, Switzerland).
9
Metabolic Profiling of Fasting Rats
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The serum concentrations of the fasting glucose (FG), total triglycerides (TG), total cholesterol (TC), and high-density lipoprotein cholesterol (HDL-C) were measured using a COBAS INTEGRA 800 auto-analyzer (Roche, Basel, Switzerland) per the manufacturer’s protocols. The fasting insulin (ml302840), glycated serum protein (GSP, ml037457), glucose kinase (GK, ml059525), glucose-6-phosphatase (G6P, ml196120), and phosphoenolpyruvate carboxy (PEPCK, ml059012) were determined using commercial kits (Shanghai Enzyme-linked Biotechnology Co., Ltd., Shanghai, China). Then, via homeostasis model assessment, the insulin resistance index (HOMA-IR) was calculated [21 (link)].
In addition, i.p. glucose tolerance tests (GTTs) were performed three days before the end of the experiment. In brief, after a 12 h fast, all rats were administered a 50% glucose solution (2.0 g/kg body weight), and blood samples from the tail veins at 0, 30, 60, 120, and 180 min were collected successively to measure the blood glucose concentration.
In addition, i.p. glucose tolerance tests (GTTs) were performed three days before the end of the experiment. In brief, after a 12 h fast, all rats were administered a 50% glucose solution (2.0 g/kg body weight), and blood samples from the tail veins at 0, 30, 60, 120, and 180 min were collected successively to measure the blood glucose concentration.
10
Blood Sample Preparation and Analysis
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The blood samples (5 mL) were collected by using Vacutainer K2E-EDTA tubes and transferred into 15 mL centrifuge tube contained Ficoll-paque PLUS medium [hall blood/Ficoll-paque PLUS medium = 4/3 (v/v)] (GE Healthcare Life Sciences, Pittsburgh, PA, USA), then centrifuged (3000 rpm, 30 min). After centrifugation, the supernatant was collected in sterile tubes and 1 N of acetic acid was added to prevent degradation. All the samples were stored at −80 °C and the storage time was less than one year. The plasma levels of fasting blood glucose (Glu-AC) and lipid profiles (TG, TCH, HDL-C, and LDL-C) were determined using the COBAS Integra 800 autoanalyzer (Roche Diagnostics, Basel, Switzerland) [25 (link),26 (link)].
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