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36 protocols using freestyle optium neo

1

Quantifying Insulin Sensitivity in Experimental Animals

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To calculate the insulin sensitivity in the experimental animals, the HOMA-index (Homeostatic Model for the Assessment of Insulin Sensitivity) was used. Assessment data are based on results from physiological studies that describe the feedback loop of glucose regulation and insulin secretion. The mathematical formula created in this way allows us to calculate the function of the beta cells and determine insulin resistance.
The index was calculated according to the following formula:
From each rat and in all groups, blood was collected following the rules for handling experimental animals. Blood glucose levels were determined from a cut to the tip of the tail of the rat and gently massaging the tail from the base upwards to generate a blood droplet. (FreeStyle Optium Neo, Abbott Diabetes Care Ltd., Maidenhead, Berkshire UK). This droplet was captured on a glucose strip placed inside a handheld glucometer (Free-Style Optium Neo, Abbott Diabetes Care Ltd., Maidenhead, Berkshire, UK), which normally requires 0.5–2 µL of blood. The result was obtained after 3 s on the glucometer screen in mmol/L. To determine insulin in the blood of animals, RAT INSULIN ELISA (BioVendor, Brno, Czech Republic) was used (the methodology of the test is described below).
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2

Capillary Blood Sampling Protocol

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Capillary blood samples were used for this portion of the study to allow standardised collection of samples in both laboratory and field conditions where metabolites were to be assessed over the entire 4‐week intervention. Finger‐tip samples were collected and immediately processed for measurement of blood lactate (Lactate Pro 2, Akray, Japan), ketones (β‐hydroxybutyrate; FreeStyle Optium Neo, Abbott Diabetes Care, Victoria, Australia) and glucose (FreeStyle Optium Neo, Abbott Diabetes Care, Victoria, Australia) concentrations. To counter any individual differences in the accuracy of these portable analysers, each participant was assigned to a specific device for the duration of their involvement in the study.
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3

Standardized Capillary Blood Analysis

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Capillary blood samples were used for this portion of the study to allow standardised collection of samples in both laboratory and field conditions where metabolites were to be assessed over the entire 5.5-wk intervention. Fingertip samples were collected and immediately processed for measurement of blood lactate (Lactate Pro 2, Akray, Japan), ß-hydroxybutyrate (FreeStyle Optium Neo, Abbott Diabetes Care, Victoria, Australia) and glucose (FreeStyle Optium Neo, Abbott Diabetes Care, Victoria, Australia) concentrations. To counter for any individual differences in the accuracy of these portable analyzers, each participant was assigned to a specific device for the duration of their involvement in the study.
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4

Glucose Sensing in Diabetic Serum

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Human serum samples were collected from diabetic patients and healthy subjects with informed consent at the First Affiliated Hospital of University of Science and Technology of China (USTC). Six human blood samples (including 4 males and 2 females based on self-report, all human subjects signed informed consent and received 200 RMB in compensation) with unknown glucose concentrations were analyzed by the same process in the section of “Gel-based sensor for glucose detection” just by replacing glucose solutions with the human serum samples. For comparison, the glucose levels in serum samples were also measured by a commercial glucometer (Abbott, FreeStyle Optium Neo, Mexico) and the blood glucose test strips (Abbott, FreeStyle Optium Neo, U.K.). All procedures are approved by Institutional Ethics Review Committee of the First Affiliated Hospital of USTC (2021 KY 089).
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5

Tail Blood Glucose Measurement

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Blood glucose levels were determined from a cut to the tip of the tail of the rat and gently massaging the tail from the base upwards to generate a blood droplet. This droplet was captured on a glucose strip placed inside a handheld glucometer (FreeStyle Optium Neo, Abbott Diabetes Care Ltd., Maidenhead, Berkshir, UK), which normally requires 0.5–2 µL of blood, and the result was obtained after 3 s on the glucometer screen in mmol/L.
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6

Exercise-Induced Blood Biomarker Sampling

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Venous blood samples were collected at pre‐exercise, post‐exercise and 1h post‐exercise into an a 6 ml sodium heparin and a 6 ml K2EDTA Vacutainer (BD, Franklin Lakes, NJ, USA) with the participants seated in an upright position. Due to the rapid exercise‐induced lymphocyte kinetics (Rooney et al., 2018 (link)), all blood samples were collected within 1 min of the specified time points. The cannula was flushed with 3–4 ml of saline every 30 min to maintain patency. Each serum vacutainer was left to clot for 30 min prior to centrifugation at 1500 g for 10 min at 4°C. After this, samples were separated into two 1.5 ml aliquots to be stored at –80°C prior to the analysis of glucose and cortisol concentration (Cobas Modular P800 Analyser, Roche Diagnostics, Auckland, New Zealand). Capillary blood d‐β‐hydroxybutyric acid (d‐βHB; Freestyle Optium Neo, Abbott Diabetes Care, Doncaster, Victoria, Australia) concentration was measured at pre‐exercise, post‐exercise and 1‐h post‐exercise from a fingertip blood sample using standardised techniques. Glucose, d‐βHB and cortisol concentrations have been previously published (Shaw et al., 2021 (link)).
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7

Measuring D-BHB in Hypoglycemic Rats

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Blood samples were obtained from the tail vein (seven animals per group) and D-BHB was measured using blood glucose and ketone monitoring system (FreeStyle Optium Neo, Abbott Diabetes Care, Limited, Witney, Oxon, UK) and keto strips (FreeStyle Optium β-ketone). Samples were obtained from intact control and fasted animals at different times throughout the experimental period. D-BHB was also determined in blood samples from animals of the Coma and the Coma + BHB groups, before (time 0) and at different times after insulin injection (1 and 2 h), at the time the animals reached the coma, at different times after recovery with glucose or glucose + D-BHB (250 mg/kg), and after the second administration of D-BHB (250 mg/kg), as indicated in Figure 2C. Animals from these groups were identically treated during the hypoglycemia period before recovery and were randomly assigned to each one of the treatments.
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8

Metabolic Profiling of Ketogenic Diet

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D-BHB and glucose were determined from blood samples taken from the tail vein (eight animals per group) using the blood glucose and ketone monitoring system (FreeStyle Optium Neo, Abbott Diabetes Care, Limited, Witney, Oxon, UK) and keto strips (FreeStyle Optium β-ketone). Data were collected from control diet and KD-fed animals before the onset of treatments and 2 weeks later when treatments were concluded.
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9

In Vitro Starch Digestion of Rice Flours

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The in vitro starch digestion method of rice samples was developed and optimized in our previous report [29 (link)], which contains detailed information about selection of digestive enzymes and glucometers. In brief, 15 mg of rice flours in triplicate were cooked with 60 µL of Milli-Q water for 20 min at 100 °C and cooled to room temperature [29 (link)]. Phosphate buffer (500 µL, pH 6.9) was added to the cooked rice and homogenized, from which 500 µL was further diluted with 1080 µL of phosphate buffer (pH 6.9) and digested with rat intestinal acetone powder for three hours. The released glucose was estimated by FreeStyle Optium Neo glucometer (Abbott Diabetes Care Ltd., Witney, UK), and rice starch digestibility is represented as mg glucose released per 100 mg dry rice flour (glucose concentration) or area under curve (AUC).
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10

Conditional Knockout Mouse Model for Usp7 and Pdx1

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All animal experiments were approved by the Francis Crick Institute and Institute of Cancer Research Animal Ethics Committees and conformed to UK Home Office regulations under the Animals (Scientific Procedures) Act 1986, including Amendment Regulations 2012. The Usp7F/F31 (link) and Pdx1-Cre49 (link) mouse lines have been previously described. These lines were inter-crossed on a C57BL/6 background to generate the genotypes of this study. All strains were genotyped by Transnetyx. The mice were housed in a constant temperature, humidity, and pathogen-free controlled environment (23 ± 2 °C, 50–60%) cage with a standard 12 h light/12 h dark cycle, plenty of water, and food (pathogen-free) in their cage. All animal experiments were conducted in adults, aged 5–8 weeks. Where indicated, blood glucose measurements were taken at the endpoint on whole blood using FreeStyle Optium Neo (Abbott) glucose meter following the manufacturer’s instructions.
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