Plasma glucose was measured in the Clinical Research Center by the glucose oxidase method (Analox Glucose Analyzer, Analox Instruments, Lunenburg, MA). For other analyses, blood samples were placed on ice at the bedside, processed within 15–20 minutes, and frozen at −80°C until the samples were analyzed. Blood samples for the analysis of drug concentration were collected prior to the morning dose, and it was assured that patients were on chronic pioglitazone and that the drug concentration was at steady-state. A novel liquid chromatography tandem mass spectrometry assay described previously was used to measure plasma concentrations of pioglitazone, hydroxypioglitazone, and ketopioglitazone.17 The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated from a single measurement of fasting insulin and glucose. Plasma insulin was determined by radioimmunoassay (Siemens, Los Angeles, CA). Adiponectin concentrations were measured by magnetic bead Milliplex technology (Luminex xMAP, Millipore, St. Charles, MO), and CK-18 levels were quantified by ELISA (M30-Apoptosense, DiaPharma, Columbus, OH).
Analox glucose analyzer
Manufactured by Analox Instruments
Sourced in United Kingdom
The Analox glucose analyzer is a laboratory instrument designed to measure the concentration of glucose in various samples. It provides accurate and reliable glucose measurements to support research, diagnostic, and monitoring applications.
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Lab products found in correlation
7 protocols using analox glucose analyzer
1
Quantification of Pioglitazone and Metabolites
2
Insulin Clamp and Glucose Kinetics
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Glucose concentration was measured every 10 min during the insulin clamp with an Analox glucose analyzer (Analox Instruments, London, U.K.). [6,6-2H2]-d -glucose enrichment in the plasma and infusate was measured using gas chromatography–mass spectrometry. As described previously, the steady-state equations of Steele et al. (29 (link)) were used to calculate the rate of glucose appearance (Ra) and disappearance (Rd). EGP was calculated as the difference between total glucose Ra and the exogenous glucose infusion rate, peripheral insulin sensitivity was assessed from the rate of glucose infusion required to maintain euglycemia during the high-dose insulin clamp, and hepatic insulin sensitivity was assessed by the extent to which EGP was suppressed from baseline to low-dose hyperinsulinemia (26 (link)).
3
Postprandial Hormonal Responses to High-Protein Meal
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Subjects will be instructed to consume the test meal within 15 minutes. Calorie intake and nutrient distribution of the meal (Boost High Protein) will be as follows: calories, 240; protein, 15 g; carbohydrates, 33 g; fat, 6 g; sodium, 200 mg; potassium, 400 mg, fiber 0 g. Patients will be able to choose between vanilla and chocolate flavors., An intravenous catheter will be inserted at 7:00 a.m. on the day of the experiment and blood will be drawn at before the meal, and at 15, 30, 60, 90, 120, and 180 minutes after the meal to measure hormonal signals of satiety. Blood samples will be collected in ethylenediaminetetraacetic acid tubes with added aprotinin (500 kallekrein inhibitory units per mL of blood) and dipeptidyl peptidase-4 inhibitor (10 μl/mL of blood; Millipore Research), and stored at –70° Celsius. Plasma concentrations of PYY, CCK, GLP-1, ghrelin, and insulin will be determined by radioimmune assay, and glucose concentration will be determined using the glucose oxidase method with an Analox glucose analyzer (Analox Instruments, Lunenburg, MA). Serum acetaminophen levels will be measured using an enzyme-linked immunosorbent assay (Abbot Laboratories, Chicago, IL).
4
Plasma Glucose and Hormone Assays
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Plasma glucose was determined at bedside by the glucose oxidase method with an Analox glucose analyzer (Analox Instruments, Lunenburg, MA). Total GLP-1 was measured by radioimmunoassay (Millipore) after plasma ethanol extraction. The assay reacts 100% with GLP17–36, GLP19–36, and GLP17–37, but not with glucagon (0.2%), GLP-2 (<0.01%), or exendin (<0.01%). Gastric inhibitory peptide (GIP) was determined by ELISA (Millipore) and reacts 100% with GIP1–42 and GIP3–42 but not with GLP-1, GLP-2, oxyntomodulin, or glucagon. Plasma insulin and C-peptide were measured by radioimmunoassay (Millipore). All hormone and metabolite assays were performed at the Hormonal Core Laboratory at the Obesity Nutrition Research Center. Intra- and interassay coefficients of variance ranged from 3.4–7.4 and 4.4–7.4%, respectively. Lipids were assayed by Ortho Clinical Diagnostics Vitros Fusion 5.1 (Ortho Clinical Diagnostics, Rochester, NY).
5
Plasma Glucose, Insulin, and C-peptide Assay
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Plasma samples were analyzed for glucose, insulin, and C-peptide levels at each time point (In Vitro Sciences Laboratory, David H. Murdock Research Institute, Kannapolis, NC). Immediately after collection, blood was centrifuged at 1300 g at 4°C for 10 minutes. The plasma was removed, and frozen at -80°C for analysis. Glucose levels (mg/dL) were obtained using an Analox Glucose Analyzer (Analox Instruments Ltd, UK) from plasma samples and a commercially available glucose kit. Insulin levels (μIU/mL) and C-peptide levels (ng/mL) were obtained using a Meso Scale Discovery Sector Imager S 600 (Meso Scale Diagnostics, LLC, Rockville, MD) with commercially available insulin and C-peptide custom kits, respectively.
6
Insulin sensitivity and ghrelin regulation
7
Comprehensive Metabolic Biomarker Profiling
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Plasma glucose was measured by the glucose oxidase method (Analox glucose analyzer; Analox Instruments, Lunenburg, MA). Plasma insulin and C-peptide concentration were measured by radioimmunoassay (Siemens, Los Angeles, CA). A1C was measured by high-performance liquid chromatography (TOSOH G-7). Plasma glucose radioactivity was measured from deproteinized plasma samples precipitated from barium hydroxide/zinc sulfate (26 (link)). Finally, total plasma adiponectin was measured by immunoassay (Milliplex MAP, EMD Millipore Corporation, Billerica, MA).
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