Prior to HFD feeding and every 4 weeks thereafter, non-fasting blood glucose was measured using a handheld glucometer (Contour Next, Bayer, Leverkusen, Germany). Fasting blood glucose measurements were collected after overnight fast using a handheld glucometer. Intraperitoneal glucose tolerance tests (IPGTTs) were performed following an overnight fast. Briefly, glucose was administered intraperitoneally at a dose of 2 g/kg body weight, then blood glucose was measured via tail vein at 0 (before glucose injection), 10, 20, 30, 60, 90 and 120 min after glucose injection using a handheld glucometer (Contour Next, Bayer). Intraperitoneal insulin tolerance tests (IPITTs) were performed following a 2 h fast. Briefly, insulin was administered intraperitoneally at a dose of 0.75 U/kg body weight, then blood glucose was measured at 0 (before insulin injection), 15, 30, 45, and 60 min after insulin injection using the handheld glucometer.
Contour next
Manufactured by Bayer
Sourced in Germany, United States, Portugal
The Contour Next is a blood glucose monitoring system designed to help individuals manage their diabetes. It features a compact and user-friendly design, allowing for convenient blood glucose testing. The device provides accurate and reliable blood glucose readings to assist patients in monitoring their condition.
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Lab products found in correlation
Ultra sensitive mouse insulin elisa kit, by Crystal Chem (4 mentions)
Elisa, by Crystal Chem (2 mentions)
Lean mass glucose, by Merck Group (2 mentions)
Echomri, by Echo Medical Systems (2 mentions)
Edta microvette, by Sarstedt (1 mentions)
β hydroxybutyrate, by Merck Group (1 mentions)
Lactate scout 4 analyzer, by EKF Diagnostics (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Sodium orthovanadate, by Merck Group (1 mentions)
Elisa, by Salimetrics (1 mentions)
K2 edta 1.0 mg microtainer tubes, by BD (1 mentions)
Humulin r u 100, by Eli Lilly (1 mentions)
Echomri device, by Echo Medical Systems (1 mentions)
Oxymax comprehensive lab animal monitoring system, by Columbus Instruments (1 mentions)
Serum separator tubes, by Sarstedt (1 mentions)
Contour xt, by Bayer (1 mentions)
Ezmadp 60k, by Merck Group (1 mentions)
Ezml 82k, by Merck Group (1 mentions)
Infinity triglyceride reagent, by Thermo Fisher Scientific (1 mentions)
Mf2100, by R&D Systems (1 mentions)
80 protocols using contour next
1
Glucose and Insulin Tolerance Assays
2
Intravenous Glucose Tolerance Test Protocol
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Fasting glucose measurements were made before administration of the experimental diet and at weeks 14 and 28 with a glucose meter (Contour Next, Bayer, Leverkusen, Germany). For the evaluation of glucose metabolism, intravenous glucose tolerance test (IVGTT) was performed as previously described [11 (link)]. Briefly, rabbits were fasted during 7 hours and then the experiment started between 14 and 15 p.m. After auricular vein cannulation, a bolus of a 60% glucose solution (0.6 g kg-1) was administered i.v. through the marginal ear vein and blood samples were taken before and at different time points after injection (15, 30, 60, 90, 120 and 180 minutes). Blood glucose was measured with a glucose meter (Contour Next, Bayer, Leverkusen, Germany). The area under the curve (AUC) was calculated by multiplying the cumulative mean height of glucose (mg dL-1) by the time (hours) [18 (link)].
3
Glucose Tolerance and Streptozotocin-Induced Diabetes
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Mice were fasted overnight prior to intraperitoneal glucose tolerance test (IPGTT), then intraperitoneal glucose was administered at a dose of 2 g/kg body weight. Blood glucose was measured using a handheld glucometer (Contour Next, Bayer, Leverkusen, Germany) via tail vein at 0 (before injection of glucose), 10, 20, 30, 60, 90 and 120 min after glucose injection. Mice were subjected to intraperitoneal administration of low dose streptozotocin (STZ, 50 mg/kg body weight) on 5 consecutive days and non-fasting blood glucose was measured via tail vein on days 0, 3, 7, 9, and 14 after starting STZ using a handheld glucometer (Contour Next, Bayer).
4
Plasma Biomarker Analysis in Rodents
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Whole-blood samples were collected from the tail into EDTA-microvettes (Sarstedt Inc) and were then centrifuged at 1000g for 10 min at 4 °C. Supernatants were aliquoted and stored at −20 °C. Plasma insulin and glucagon were determined by ELISA (Crystal Chem). Plasma free fatty acids (FFAs) (Abcam) and β-hydroxybutyrate (Sigma-Aldrich) were measured using a commercial kit.
Blood glucose and lactate levels were measured using a glucometer (Bayer Contour Next, Bayer Healthcare) and a lactometer (Lactate Scout 4 analyzer, EKF Diagnostics) with tail-tip bleeding.
Blood glucose and lactate levels were measured using a glucometer (Bayer Contour Next, Bayer Healthcare) and a lactometer (Lactate Scout 4 analyzer, EKF Diagnostics) with tail-tip bleeding.
5
Salivary Biomarker Analysis Protocol
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Saliva was processed within one hour of collection. Protease inhibitor cocktail (Sigma Aldrich, Atlanta, GA) 2 μL/mL of saliva and sodium orthovanadate (400mM, Sigma Aldrich, Atlanta, GA) 3 μL/mL of saliva was added to each sample that was then centrifuged at 4°C and 1500xg for 15 minutes. Saliva samples were then divided into 0.5 mL aliquots and frozen at -80°C until assayed. The pellet collected during centrifugation was frozen at -80°C until processed for microbiome analysis (see below).
Salivary samples were assayed for acidity using short-range pH paper (range 5.5–8.0 with 0.2 increments). Salivary glucose levels were measured using a Bayer Contour Next glucometer (Bayer Healthcare, Mishawaka, IN). The detection limit of the meter is 10 mg/dL. Duplicate samples were assayed for nitric oxide using Griess reagent [28 (link)] and for CRP and IL-1β by ELISA (Salimetrics LLC, State College, PA). Lower limits of detection for ELISA were 0.43 pmol/L for CRP and 0.022 pmol/L for IL-1β.
Salivary samples were assayed for acidity using short-range pH paper (range 5.5–8.0 with 0.2 increments). Salivary glucose levels were measured using a Bayer Contour Next glucometer (Bayer Healthcare, Mishawaka, IN). The detection limit of the meter is 10 mg/dL. Duplicate samples were assayed for nitric oxide using Griess reagent [28 (link)] and for CRP and IL-1β by ELISA (Salimetrics LLC, State College, PA). Lower limits of detection for ELISA were 0.43 pmol/L for CRP and 0.022 pmol/L for IL-1β.
6
Chronic Stress Blood Biomarkers Analysis
7
Insulin Sensitivity Measurement in Mice
8
Metabolic Phenotyping in Mice
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Glucose and insulin tolerance test: Glucose and insulin tolerance tests were performed in 12 mos mice, following 10 mos on the WD. After fasting for 5 hours, glucose tolerance test (GTT) was performed (Agri Laboratories, LTD). For the insulin tolerance test (ITT), mice received a single intraperitoneal injection of a diluted insulin solution (males 0.75 IU/kg and females 0.50 IU/kg; prepared in sterile saline) (Humulin R U-100, Eli Lilly). Glucose was determined using a handheld glucometer (Bayer Contour Next).
NMR Body Composition: Body composition was determined in the Echo MRI 3-in-1, time domain nuclear magnetic resonance (TD-NMR) system (Houston, TX). Body weight was measured on a standard laboratory balance before mice were placed in to a clear, cylindrical holder. The tube was gently inserted in to the boor for an approximately two-minute measurement. Adiposity (% body fat) was calculated as ((fat mass/total body weight) x 100).
Plasma lipid measurements: Blood was collected in K2 EDTA (1.0mg) microtainer tubes (BD) at harvest (non-fasted) and kept at room temperature for at least 30 minutes to prevent clotting and then centrifuged at 22°C for 15 minutes at 5000rpm. Plasma was carefully collected and aliquoted. Plasma was characterized on the Beckman Coulter AU680 chemistry analyzer.
NMR Body Composition: Body composition was determined in the Echo MRI 3-in-1, time domain nuclear magnetic resonance (TD-NMR) system (Houston, TX). Body weight was measured on a standard laboratory balance before mice were placed in to a clear, cylindrical holder. The tube was gently inserted in to the boor for an approximately two-minute measurement. Adiposity (% body fat) was calculated as ((fat mass/total body weight) x 100).
Plasma lipid measurements: Blood was collected in K2 EDTA (1.0mg) microtainer tubes (BD) at harvest (non-fasted) and kept at room temperature for at least 30 minutes to prevent clotting and then centrifuged at 22°C for 15 minutes at 5000rpm. Plasma was carefully collected and aliquoted. Plasma was characterized on the Beckman Coulter AU680 chemistry analyzer.
9
Insulin Tolerance Test in Mice
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At 6 weeks post AAV injection, mice were injected intraperitoneally with an insulin solution (0.5 U of insulin per kilogram body weight) following a 2-h fast. Tails were cut (<0.5 mm) with surgical scissors to elicit blood flow. Blood was obtained from the tail at baseline, 15, 30, 60, 90, and 120 min after insulin injection. Blood glucose concentrations were measured with a portable glucose meter using default manufacturer settings (Bayer Contour Next). Following blood collection, styptic powder was used to stop bleeding.
10
Postprandial Glucose Monitoring Protocol
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Capillary blood glucose measures were collected and assessed by using reactive strips and glucose monitors (Contour®Next, Bayer HealthCare S.p.A., Milan, Italy), previously validated [36 (link),37 (link)]. Two measures were performed at each time point, and the average of these was considered. In case of a difference greater than 10% between the two measures, a third measure was performed. As schematically shown in Figure 1 , glycemic measures were collected immediately before the meal, every 15 min after the meal consumption for the first two hours, during which glycemic concentration in the blood is higher than baseline levels, and thus it requires more frequent measurements in order to evaluate the post-meal fluctuations; during the third hour, when glycemia has been previously shown to return to baseline levels, it was measured every 30 min [38 (link)]. Participants measured glycemia while sitting, and before each measure, they were requested to wash their hands to avoid possible alterations due to external factors.
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