Equal numbers of male and female mice were fasted for 6 hours then injected with [3H]-2DG (20uCi/mouse; ip). Blood samples were taken from the tail vein at 30, 60, 90 mins post-injection to determine blood glucose levels using a Nova Max Plus glucometer according to manufacturer’s instructions (Nova Biomedical Corporation, Waltham, MA) to determine [3H]-2DG specific activity. After the final collection of blood, mice were euthanized with pentobarbitol (i.p. 60mg/kg body weight) and the heart and brain removed, rinsed in PBS, diced and frozen in dry ice. Tissues were processed with perchloric acid, barium hydroxide/zinc sulfate and glucose uptake rate was calculated according to the method of Ferre’, et al (28 (link)).
Nova max plus
Manufactured by Nova Biomedical
Sourced in United States, Switzerland
The Nova Max Plus is a diagnostic device used to measure blood glucose levels. It provides quantitative results through a fingerstick blood sample.
Automatically generated - may contain errors
Lab products found in correlation
Uv visible spectrophotometer, by Beckman Coulter (1 mentions)
Nefa hr 2 kit, by Fujifilm (1 mentions)
Infinity kit, by Thermo Fisher Scientific (1 mentions)
Total cholesterol e kit, by Fujifilm (1 mentions)
Lactate plus meter, by Nova Biomedical (1 mentions)
Microvette cb 300, by Sarstedt (1 mentions)
Ultra fine nano pen needles, by BD (1 mentions)
Accu check nano, by Roche (1 mentions)
10 protocols using nova max plus
1
Glucose Uptake Measurement in Mice
2
Intraperitoneal Glucose Tolerance Test
3
Glucose, Lactate, and Perceived Exertion
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Blood glucose was measured using finger sticks and an over-the-counter glucometer (Nova Max Plus, Nova Biomedical Corp.). At each reading (baseline and minutes 15, 30, 45, and 60) multiple measurements were taken until two readings within 15 mg/dL were obtained and subsequently averaged as previously described [10, 17] .
Blood pressure and heart rate were measured at the same time points using an automated blood pressure monitor (Omron 3 series).
Lactate was measured from intravenous blood samples collected at baseline and minute 30 and analyzed with a Biosen C-Line Clinic glucose and lactate analyzer (EKF Diagnostics).
Ratings of perceived exertion were obtained using the modified Borg scale at the midpoint and end of each SCD bout.
Blood pressure and heart rate were measured at the same time points using an automated blood pressure monitor (Omron 3 series).
Lactate was measured from intravenous blood samples collected at baseline and minute 30 and analyzed with a Biosen C-Line Clinic glucose and lactate analyzer (EKF Diagnostics).
Ratings of perceived exertion were obtained using the modified Borg scale at the midpoint and end of each SCD bout.
4
Bovine Blood Sampling and Analysis
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Blood samples were obtained from the jugular vein using a 20-gauge needle before the administration of treatments. Once each heifer was assigned to her respective treatment, blood samples were collected every Tuesday at 0800 h for the duration of the study. Samples were collected in two 10-mL evacuated tubes, the first containing EDTA anticoagulant and the second without anticoagulant (Monoject, Covidien Ilc., Mansfield, MA). Blood ketone concentrations were obtained using a hand-held electronic blood glucose and ketone monitoring device (Nova Max Plus, Nova Biomedical, Waltham, MA; Deelen et al., 2016) (link). A whole-blood sample, not containing EDTA, was transferred to the sensor of the test strip using a disposable pipette.
Samples with EDTA were placed on ice until they were centrifuged at 1,278 × g at 4°C for 20 min (5430R, Eppendorf, Hamburg, Germany). Plasma was stored in 2 aliquots and frozen at -20°C until further analysis of plasma urea nitrogen (PUN) and glucose. Urea concentrations were measured in duplicate using the diacetyl-monoxime method and measured colorimetrically using a UV-visible spectrophotometer (Beckman Coulter Inc., Brea, CA) set at a wavelength of 540 nm. Plasma glucose concentrations were measured in duplicate via Wako Autokit for Glucose (Wako Diagnostics, Mountain View, CA) and read on a UV-visible spectrophotometer at a wavelength of 505 nm.
Samples with EDTA were placed on ice until they were centrifuged at 1,278 × g at 4°C for 20 min (5430R, Eppendorf, Hamburg, Germany). Plasma was stored in 2 aliquots and frozen at -20°C until further analysis of plasma urea nitrogen (PUN) and glucose. Urea concentrations were measured in duplicate using the diacetyl-monoxime method and measured colorimetrically using a UV-visible spectrophotometer (Beckman Coulter Inc., Brea, CA) set at a wavelength of 540 nm. Plasma glucose concentrations were measured in duplicate via Wako Autokit for Glucose (Wako Diagnostics, Mountain View, CA) and read on a UV-visible spectrophotometer at a wavelength of 505 nm.
5
Measuring Glucose and Ketone Levels
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In both rats and mice, tail vein blood was collected and glucose levels were measured with a human glucometer (Nova Max Plus, Nova Biomedical Corporation). Serum was separated from the same blood sample to quantify β-hydroxybutyrate levels using a colorimetric assay kit (Cayman Chemical, Item Number 700190).
6
Measuring Tissue Glucose Uptake in Mice
7
Metabolic Profiling of Fasted Mice
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PcxL−/− and Pcxf/f mice were fasted at indicated time points and blood glucose and lactate levels were measured using a Nova Max Plus glucometer and Lactate Plus meter (Nova Biomedical). Blood was obtained by tail vain bleeds and collected using capillary tubes (Microvette CB 300, Sarstedt) and serum was isolated from blood following manufacturer’s instructions and stored at −80 °C. Serum TGs were analyzed using an Infinity Kit (Thermo Fisher Scientific). Serum cholesterol was assessed using a total Cholesterol E Kit (FUJIFILM Wako). Serum NEFAs were measured using a NEFA HR(2) kit (FUJIFILM Wako). Serum BHB was measured using a LiquiColor assay (Stanbio Laboratory). Biological replicates were used for all assays (n = 6–14/group).
8
CRN02481 Oral Dosing in Sur1−/− Mice
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Sur1−/− mice and WT mice received CRN02481 orally at a dose of 30 mg/kg or an equal volume of vehicle (25 mM Citrate Buffer, pH 3) after an overnight fast (16–18 h). This dose was selected based on a short-term dose-finding study where the effect of different doses on fasting glucose was evaluated (57 (link)). A copy of the poster can be found here: https://crinetics.com/pipeline/crn04777-oral-sst5-agonist-congenital-hyperinsulinism-congenital-hi/ ). Plasma glucose and β-hydroxybutyrate levels (BHB) were assessed at baseline (0 h), 1, and 2 h after treatment by a portable glucose meter (Stat Strip Xpress Glu, Nova biomedical) and Ketone meter (Nova Max plus, Nova biomedical). Blood was obtained from a tail nick. Blood was collected for measurement of plasma insulin levels using Mouse Ultrasensitive Insulin ELISA kit (Catalog # 80-Insmsu-E10, Alpco, RRID:AB_2792981 ).
9
Microneedle-Assisted ISF Extraction and Measurement
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ISF was extracted using our previously published methods [8 (link),9 (link),10 (link)]. Briefly, animals were anesthetized using 2% isoflurane. Microneedle arrays were then applied to extract ISF. Ultra-fine Nano PEN needles (BD, Franklin Lakes, NJ, USA) were placed into 3D-printed microneedle array holders. Each needle was attached to a 1–5 µL calibrated pipet capillary tube (Drummond Scientific Co., Broomall, PA, USA). The array assembly was then pressed into the dermal tissue of the rats and held in place until enough ISF was collected. ISF was transferred from the capillary tubes onto the glucose or ketone meter strips using a small plunger that is supplied with the capillary tubing. Glucose and ketone measurements were immediately recorded using a Nova Max Plus glucose/ketone meter (Nova Biomedical, Waltham, MA, USA) with Nova Max Plus glucose and ketone test strips (Nova Biomedical, Waltham, MA, USA), and another ISF extraction was then performed. This routine was repeated for 1.8 h for multiple glucose and ketone measurements of both blood and ISF. The blood was obtained from a small tail snip, where a drop of blood from the tail was periodically placed on the glucose and ketone test strips to measure the blood concentrations correlating with the times of ISF measurement. All animals had a terminal cardiac puncture performed at the conclusion of each experiment.
10
Measuring Blood Glucose and Ketones in Sepsis
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