Blood samples were obtained after 8–12 h fast. Cholesterol, triglycerides, HDL cholesterol and apolipoprotein B were measured in serum using colorimetric assays (Unicel DxC 600 Synchron Clinical System Beckman Coulter). VLDL lipoproteins were isolated using sequential ultracentrifugation (Optimal Beckman LE80-K) of 40,000 RPM at 4 °C for 18 h. Serum aliquots (3.5 mL) were centrifuged at background density of 1.006 Kg/L, VLDL-C and VLDL-triglycerides levels in the ultracentrifugal bottom fraction were analyzed by calorimetric assays (Unicel DxC 600 Synchron Clinical System Beckman Coulter). LDL and VLDL cholesterol were calculated using the Friedewald’s equation (VLDL-F, LDL-F), Sampson’s method (VLDL-S, LDL-S) and the calculation proposed by Martin et al. (VLDL-M, LDL-M). LDL-C was also calculated using VLDL-C measures by ultracentrifugation to approximate a gold-standard for comparative assessments.
Unicel dxc 600 synchron clinical system
Manufactured by Beckman Coulter
Sourced in United States
The Unicel DxC 600 Synchron Clinical System is a fully automated clinical chemistry analyzer designed for high-volume clinical laboratories. It is capable of performing a wide range of biochemical tests on patient samples to aid in clinical diagnosis and treatment monitoring.
Automatically generated - may contain errors
Lab products found in correlation
Access 2, by Beckman Coulter (2 mentions)
Colorimetric determination reagent, by Fujifilm (2 mentions)
Meso scale discovery kits, by MSD (2 mentions)
Sector imager 2400, by MSD (2 mentions)
Elisa kit, by R&D Systems (1 mentions)
Unicel dxi 800 access immunoassay system, by Beckman Coulter (1 mentions)
Le 80k, by Beckman Coulter (1 mentions)
Variant 2 turbo, by Bio-Rad (1 mentions)
Unicel dxh 800 coulter cellular analysis system, by Beckman Coulter (1 mentions)
1 ml microcentrifuge tube, by Eppendorf (1 mentions)
21g needle, by BD (1 mentions)
Elecsys 2010 immunoassay analyzer, by Roche (1 mentions)
Architect clinical chemistry analyzer, by Abbott (1 mentions)
Paxgene blood rna tube, by Preanalytix (1 mentions)
Variant 2 turbo system, by Bio-Rad (1 mentions)
Dca vantage analyzer, by Siemens (1 mentions)
Access 2 immunoassay system, by Beckman Coulter (1 mentions)
D 10 hemoglobin testing system, by Bio-Rad (1 mentions)
19 protocols using unicel dxc 600 synchron clinical system
1
Measuring Serum Lipid Profiles
2
Metabolic Biomarkers for MS Cohort
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Plasma glucose concentration was measured by an automated glucose analyzer (Yellow Springs Instruments Co.), serum insulin concentration was measured by using a chemiluminescent immunoassay (Beckman Coulter Access 2). Lipid concentrations (cholesterol, triglycerides, and HDL cholesterol), and ApoB, were measured using colorimetric assays (Unicel DxC 600 Synchron Clinical System Beckman Coulter). LDL-cholesterol was calculated using Martin’s equation. HOMA2-IR was calculated using fasting glucose and insulin using the HOMA2 calculator released by the Diabetes Trials Unit, University of Oxford: HOMA Calculator; IR was defined as a HOMA-IR value >2.5, as previously shown for the MS cohort.11 (link) ApoB >90th percentile was evaluated as a risk factor according to previously validated cut-off value in Mexican population.12 (link)
3
Comprehensive Biomarker Evaluation Protocol
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Blood glucose and lactate (LA) were evaluated with YSI instruments (YSI 2300 and 1500, Yellow Spring Instrument Co. Ltd., Greene County, OH, USA), respectively. Serum insulin and cortisol were evaluated by the immunofluorescence assay on the Unicel® DxI 800 Access® Immunoassay System and also with evaluation kits provided by the same manufacturer (Beckman Coulter, Brea, CA, USA). The reference range for the insulin kit was 0.21–2100 pmol/L. Intra-assay coefficient of variation (CV) was 2.0%–4.2% and inter-assay CV was 3.1%–5.6%. Assay sensitivity for the cortisol kit was 11.0 nmol/L. Intra-assay CV was 4.4%–6.7%, and an inter-assay CV was 6.4%–7.9%. Serum glucagon was evaluated with an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems Inc., Minneapolis, MN, USA) according to the instructions provided by the manufacturer. Sensitivity for the glucagon kit was 14.7 pg/mL and reference range was 31.3–2000 pg/mL. Intra-assay CV was 2.7%–3.6%, and inter-assay CV was 5.8%–8.7%. Serum free fatty acid (FFA) was evaluated by the Randox method (Range 0.072–2.24 mmol/L; CV < 5%) on the UniCel® DxC 600 Synchron® Clinical System (Beckman Coulter, Brea, CA, USA).
4
Anthropometric Assessment and Metabolic Markers
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We calculated the body mass index using anthropometric evaluation using the formula of weight in kilograms divided by height in m2. Blood samples were acquired after a 10–12 h fast to measure fasting glucose (Yellow Springs Instruments Co.); serum lipid concentrations assessed total cholesterol, triglycerides and HDL-C and were measured using colorimetric assays (Unicel DxC 600 Synchron Clinical System Beckman Coulter).
5
Biomarker Measurement in Fasting Samples
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Information about personal and medical factors was obtained using standardized questionnaires. Prior to the intervention, fasting venous blood samples were collected in EDTA containers, immediately centrifuged and stored at -80°C. Serum creatinine, cystatin C, high-sensitivity C-reactive protein, and interleukin 6 (IL-6) were measured on a Beckman Coulter Unicel DxC600 Synchron Clinical System (Beckman Coulter, California). High-sensitivity troponin T (cTnT) and N-terminal proB-type natriuretic peptide (NT-proBNP) were assessed on the Elecsys 2010 immunoassay analyzer (F. Hoffmann–La Roche, Switzerland). The body surface was estimated by using Du Bois’s equation (Body surface area (m2) = [Weight (kg)0.425 x height (m)0.725] x 0.007184). The creatinine and cystatine C based chronic kidney disease epidemiology collaboration (CKD-EPI) formula was applied to estimate the glomerular filtration rate (eGFR).[17 (link)]
6
Comprehensive Metabolic Profiling in Diabetes
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Age, sex, type and duration of diabetes mellitus, body mass index (BMI [calculated as the weight in kilograms divided by the squared height in centimeters]), were collected from the clinical record. Metabolic parameters including uric acid, creatinine, lipid profile (total cholesterol, high density cholesterol [HDL-C] and triglycerides), apolipoprotein B (Apo B), and hepatic enzymes (AST, ALT) were measured with colorimetric assays (Unicel DxC 600 Synchron Clinical System Beckman Coulter). Low density cholesterol (LDL-C) as calculated using the Martin's formula [23 (link)]. Glycosylated hemoglobin (HbA1c) concentration was assessed by HPLC (Variant II Turbo, Bio-Rad). Glomerular filtration rate (eGFR) was estimated using the Cockroft-Gault equation and the urinary creatinine to albumin ratio (UACR) was also calculated.
7
Lipid and Blood Profile Analysis
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Fasting plasma concentrations of glucose, total cholesterol, LDL + very low-density lipoprotein (VLDL) cholesterol (LDL + VLDL), high-density lipoproteincholesterol (HDL), and triglycerides were measured using an Unicel DxC 600 Synchron Clinical System (Beckman Coulter). The normal range of total cholesterol was anticipated to be 53–146 mg/dL, and the normal HDL levels in these animals have been reported previously to be approximately 62 mg/dL (25 (link)). LDL + VLDL was calculated by subtracting HDL from total cholesterol levels for each animal. Complete blood counts were determined using an Unicel DxH 800 Coulter® Cellular Analysis System (Beckman Coulter).
8
Quantification of Liver Enzymes in Mice
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Blood samples were collected 1.5 or 24.5 h after the first treatment and were collected by terminally anaesthetising the mice using isofluraneand performing a cardiac puncture with a 21G needle (Becton Dickinson, Franklin Lakes, NJ, USA) and a heparinised syringe. A volume of approximately 600 µl was collected from each mouse into a 1-ml microcentrifuge tube which had been washed with heparin (Eppendorf, Hamburg, Germany). To separate the serum, the samples were centrifuged at 3,000 rpm at 4°C for 15 min and 300 µl of the supernatant was subsequently transferred to a 1-ml microcentrifuge tube (Eppendorf). Sampled were stored at -20°C until analyzed.
Two liver enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were quantified using a UniCelDxC 600 Synchron Clinical System (Beckman Coulter, Brea, CA, USA) following the manufactures recommend procedures and reagents.
Literature values for the normal range in mice were used to minimize the number of animals used. Specifically, the normal range for AST and ALT in mice are reported to be in the range of 300 ± 100 units/L and 100 ± 50 units/L (Oršolić et al., 2010 (link); Gao et al., 2014 (link)).
Two liver enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were quantified using a UniCelDxC 600 Synchron Clinical System (Beckman Coulter, Brea, CA, USA) following the manufactures recommend procedures and reagents.
Literature values for the normal range in mice were used to minimize the number of animals used. Specifically, the normal range for AST and ALT in mice are reported to be in the range of 300 ± 100 units/L and 100 ± 50 units/L (Oršolić et al., 2010 (link); Gao et al., 2014 (link)).
9
Standardizing Urine Metabolomic Profiling
10
Cardiovascular Biomarker Measurement Protocol
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Prior to ECV and at follow-up, venous blood samples were collected in EDTA tubes and PAXgene™ Blood RNA tubes (PreAnalytiX). EDTA tubes were immediately centrifuged to isolate plasma and all tubes were stored at -80°C. High-sensitivity C-reactive protein (hs-CRP), cystatin C (CYSC), and interleukin-6 (IL6) were measured on a Beckman Coulter Unicel DxC600 Synchron Clinical System (Beckman) according to the manufacturer’s protocol. Myeloperoxidase (MPO) was measured using the ARCHITECT MPO immunoassay on the ARCHITECT Clinical Chemistry Analyzer (Abbott). N-terminal pro B-type natriuretic peptide (NT-proBNP) was measured on the Elecsys 2010 immunoassay analyzer (Roche).
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