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41 protocols using vitros 5600

1

Multiparametric Biomarker Assessment in AECOPD-PH

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WBC, RBC, RDW, and MPV (XN-2000, SYSMEX, Japan). ALB, TG, TC, HDL, LDL and TBIL (Cobas8000, Roche, Germany). FDP and DD (ACL-TOP-750, Werfen, Spain). NT-ProBNP (VITROS 5600, Ortho Clinical Diagnostics, American). CRP (VITROS 5600, Ortho Clinical Diagnostics, American), IL-6 and PCT (Cobas701, Roche, Germany). Echocardiography was performed in AECOPD-PH patients (EPIQ-7C, Philips, Netherlands).
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2

COVID-19 Glucose and Lipid Metabolism

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SARS-CoV-2 infection was diagnosed based on clinical symptoms and confirmation by RT-PCR. Fasting glucose levels were measured by dry chemistry with the colorimetric method (Vitros 5600; Ortho Clinical Diagnostics), and pre- and postprandial glucose levels were measured by the capillary method using an Accu-Chek glucometer. Hypoglycemia was defined as a glucose level ˂70 mg/dl (3.88 mmol/l). Glycated hemoglobin A1C (HbA1C) values were determined using high-performance liquid chromatography with a DS-5 Analyzer (Drew Scientific, Inc., Miami, FL, USA). Lipid levels were measured by dry chemistry with the colorimetric method (Vitros 5600, Ortho Clinical Diagnostics), C reactive protein (CRP) levels were measured by a chemiluminescent immunometric assay (Vitros 350), D dimer levels were measured by fluorescence immunoassay (Quidel Cardiovascular Inc., CA USA) and fibrinogen levels were measured by the Clauss method (Instrumentation Laboratory Company, Bedford MA, USA).
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3

Hormonal Biomarkers Quantification Protocol

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GH was measured by a solid-phase, two-site chemiluminescent immunometric assay (Immulite 2000, Siemens, Berlin, Germany), plasma glucose by an oxidase colorimetric reaction (Vitros 5600, Ortho Clinical Diagnostics, New Jersey, USA) and betahydroxybutyrate by a D-3 hydroxybutyrate dehydrogenase colorimetric reaction (Vitros 5600, Ortho Clinical Diagnostics, New Jersey, USA). IGF-I was measured by radioimmunoassay after acid ethanol extraction (Esoterix Laboratories, Texas, USA) and IGFBP-3 was measured by radioimmunoassay (Esoterix Laboratories, Texas, USA). Where relevant, assay-specific Z-scores for age and gender were reported.
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4

Biochemical Measurements in Clinical Trial

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Glucose was measured with Analox glucoanalyzer GM9 and colorimetric glucose oxidase (Vitros 5600; Ortho Clinical Diagnostics). Lipid levels were measured by dry chemistry with colorimetric method (Vitros 5600; Ortho Clinical Diagnostics). Insulin (μU/ml) and C-peptide (ng/ml) were measured by chemiluminescent immunometric assay (IMMULITE 2000 Immunoassay system, Siemens). HbA1c was determined using high-performance liquid chromatography with DS-5 Analyzer (Drew Scientific, Inc. Miami, FL, USA). Personnel performing measurements were blinded to treatment allocation7 (link).
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5

Comprehensive Metabolic Profile Assessment

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HbA1c, serum total cholesterol, serum high-density lipoprotein (HDL) cholesterol, serum triglycerides, and plasma creatinine were measured from non-fasting venous blood samples and collected on the examination day. HbA1c was analysed by high-performance liquid chromatography on a Tosoh G7 (Tosoh Corporation, Tokyo, Japan). Triglycerides, HDL, and total cholesterol were analysed by standard enzymatic colorimetry techniques on a Vitros 5600 (Ortho Clinical Diagnostics, Illkirch-Graffenstaden, France). Serum LDL cholesterol was calculated using the Friedewald equation.
All biochemical measures were analysed in the laboratory at Steno Diabetes Center, Denmark, except for the plasma samples used for the analysis of AGEs and dicarbonyls. The samples were stored at −80°C at Steno Diabetes Center and transferred on dry ice to the laboratory at the University of Heidelberg, Germany, for analysis.
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6

Solubility Determination of T3 Hormone

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3,3′,5-Tri-iodo-l-thyronine (T3, #T2877; Sigma-Aldrich) was weighed (0.003 g) on an analytical scale, dissolved in 20 ml sterile PBS, heated at 37°C for 10 min, and vortexed at the highest setting for 1 min. As a percentage of T3 remains insoluble using this method, we determined the resulting free T3 of this preparation compared with alkalizing T3 with 1 N NaOH (1 mL per mg) and putting in PBS, as described by the manufacturer. Analysis of free T3 heated/vortexed vs NaOH solubilized was made on the Vitros 5600 chemistry analyzer (Ortho Clinical Diagnostics, Rochester, NY, USA) in a matrix-compatible diluent to a final concentration of 50 ng/mL. With NaOH solubilization and dilution in PBS, 52.3 ± 8.4 ng/mL (N = 3) was recovered compared with 0.87 ± 0.012 ng/mL (N = 3). The recovery ratio (60.4 = 52.3/0.87) was used to estimate the concentrations of T3 in all the experiments described in vitro and in vivo.
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7

Noninvasive Serum Biomarkers for NAFLD

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The biomarker determinations were performed using commercially available laboratory kits (XT-1800i, Sysmex for platelets; Vitros 5600, Ortho-Clinical Diagnostics, Johnson & Johnson for biochemical parameters). The noninvasive serum biomarker analysis included APRI and FIB-4, which were calculated according to the published analytic recommendations [6 , 7 (link)], as follows: APRI=ASTIU/L/ASTULNIU/L/plateletcount109/L×100;FIB4=ageyears×ASTIU/L/plateletcount109/L×ALTIU/L
Additionally, modified APRI (M-APRI) and modified FIB-4 (M-FIB-4) were calculated by including BMI z-scores in both formulas, as follows: MAPRI=BMIzscore×APRI;MFIB4=BMIzscore×FIB4.
A novel simple biomarker, B-AST = BMI z-score × AST (IU/L), was also proposed and calculated.
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8

Biochemical Markers Assessment Protocol

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Levels of creatinine, urea, total bilirubin, and lactic acid were determined in mg dL–1 and processed on a Vitros 5600 (Ortho Clinical Diagnostics, Raritan, NJ, USA) analytical platform, using the dry chemistry method.
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9

Estimating Kidney Function from Frozen Samples

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Frozen samples were used to estimate kidney function. Blood stored at the laboratory at the Steno Diabetes Center, Copenhagen, at −80°C from the years 1999–2001 and 2005–2010 was analysed for creatinine levels using “Vitros 5600” Ortho Clinical Diagnostics [22 ]. Estimated glomerular filtration rate (eGFR) was calculated using serum creatinine values expressed as millilitres per minute and adjusted for mean body surface area of 1,73 m2, age and sex according to the CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration) formula with CKD cut-off at eGFR <60 ml/min/1.73 m2. We used Danish guidelines [23 ] similar to the 2012 KDIGO guidelines (Kidney Disease: Improving Global Outcomes) defining albuminuria as urine albumin creatinine ratio in a random spot urine >30 mg/g [24 ].
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10

Comprehensive Metabolic Profiling Protocol

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All laboratory measurements were obtained the day after the liver MRI, following a 12 hour fast. Analyses were performed by the University of Colorado Anschutz Research core laboratory or the Children’s Hospital Colorado clinical laboratory. Plasma total cholesterol, high-density lipoprotein cholesterol (HDL-C) (Hitachi 917 autoanalyzer; Boehringer Mannheim Diagnostics, Indianapolis, IN) and triglycerides (Beckman Coulter, Brea, CA) were analyzed enzymatically. Insulin was analyzed with radioimmunoassay (Millipore, Billerica, MA). Plasma glucose was measured at the bedside using the StatStrip® Hospital Glucose Monitoring System (Novo Biomedical, Waltham, MA, USA). Hemoglobin A1c (HbA1c), was measured with HbA1c immunoassay analyzer (Siemens DCA Vantage, Siemens Medical Solutions, CA). Alanine aminotransferase (ALT) was determined via VITROS 5600 (Ortho Clinical Diagnostics, Rochester, NY) and was obtained the day of the screening visit, within 1 month of the MRI. SHBG was measured using an electrochemiluminescence immunoassay (Esoterix, Calbassas Hills, CA). Total testosterone was analyzed using a liquid chromatography–tandem mass spectrometry and free testosterone with equilibrium dialysis (Esoterix, Calbassas Hills, CA). Hepatic insulin resistance was estimated using HOMA-IR and calculated as (fasting plasma glucose [mg/dL] x fasting plasma insulin [mg/dL] / 405) (28 (link)).
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