Accu chek test strips

Mtbp^+/− [29 (link)] and littermate-matched Mtbp^+/+ C57Bl/6 mice of both genders were generated through interbreeding and were housed together. For survival analysis, mice were sacrificed after meeting humane end-of-life criteria. Necropsy with tissue collection was performed and tissues were evaluated by a board-certified veterinary pathologist (K.B.) in a blinded manner. For analysis of healthy aged mice (29 months-old), the mice were starved for 5 hours and crown-to-rump length was measured. Blood was collected and analyzed for blood glucose levels with Accu-chek test strips (Roche Diagnostics, Indianapolis, IN, USA; [41 (link)], centrifuged and plasma frozen for later analysis. Mice were sacrificed by cervical dislocation. Liver and gastrocnemius muscle were frozen with a Wallenburg clamp pre-cooled in liquid nitrogen as previously described [42 (link)]. Brown fat pads were collected and frozen. Tissues were kept at −80°C until analysis. Femurs were collected and measured with electronic calipers. Experiments were approved by the Vanderbilt Institutional Animal Care and Use Committee and followed all federal and state rules and regulations.

Animal experiments were approved by the Animal Care and Use Committee of Sunnybrook Research Institute (AUP #467) in Toronto, ON. The National Institutes of Health Guidelines for the Care and Use of Experimental Animals were met. 6-week-old male C57BL/6 mice were purchased from The Jackson Laboratory (ME, USA) and were randomly chosen to receive HFD (TD.06414, Harlan Laboratories, WI, USA) to induce obesity; mice were fed low-fat diet (LFD, TD.08806, Harlan Laboratories, WI, USA) as control. After 16 weeks of feeding, IPGTT was performed by intraperitoneal injection of 20% glucose solution (2 g glucose per kg body weight) after overnight fasting followed by blood glucose measurement (Accu-Chek test strips, Roche, USA) at 0, 15, 30, 60 and 120 min after the glucose intraperitoneal injection. The animals in each group were sub-divided into sham and burned groups (N = 6 in each group). HFD/LFD and water was given ad libitum upon arrival until sacrifice. The animals were randomized into 4 groups: LFD sham, HFD sham, LFD burn, and HFD burn. A well-established method was used to induce a full-thickness scald burn of 20% TBSA^{45 (link)}. Second IPGTT was performed on post-burn day 6 and all the animals were sacrificed on post-burn day 7.

Eight‐week‐old male C57BL/6J mice (n = 45, weight 25‐30 g) were obtained from the Animal Center of Chongqing Medical University. Mice were housed in a specific pathogen‐free mouse facility on a 12‐hour light‐dark cycle, with ad libitum access to food and water. Mice were randomly divided into three groups (n = 15 per group): Control group, DM group and DM+ FPS‐ZM1 group. Diabetic mice were induced as previously reported with some modifications,34 after a 12‐hour fast, mice received a single 150 mg/kg intraperitoneal injection of STZ (Sigma, St. Louis, MO, USA). Control mice received an equivalent volume of 0.9% saline injection. Fasting blood glucose was measured 3, 7, 10 and 20 days after STZ injection using ACCU‐CHEK Test Strips (Roche Ltd). Mice with fasting blood glucose above 300 mg/dL were considered as diabetic model. FPS‐ZM1 which can cross the blood‐brain barrier is the high‐affinity RAGE‐specific blocker. As previously described with some modifications, 3 months after diabetes induction, mice in the DM+FPS‐ZM1 group received 1 mg/kg/d intraperitoneal injection of FPS‐ZM1 (0.1 mg/mL) (Cayman Chemical, USA) and mice in other groups received an equivalent volume of 0.9% saline injection for 4 weeks.34, 35, 36, 37 Then all mice were killed.

An OGTT was performed during the last week of the study. After being subjected to 14 h of fasting, the mice were administered oral glucose (2 g/kg), and blood was obtained from the tail vein 0, 30, 60, and 120 min after glucose treatment. Glucose levels (mg/dL) were measured using Accu-Chek test strips on an Accu-Chek Active blood glucose meter (Roche Diagnostics, Rotkreuz, Switzerland). The glucose area under the curve was calculated by plotting the glucose concentration as a function of time (min).

We induced diabetes by a single i.p. injection of freshly prepared streptozotocin (STZ) (150 mg/kg; Sigma-Aldrich, Germany) in sterile 0.1 M sodium citrate buffer (pH 4.5) [33 (link)]. Control mice were only injected i.p. with vehicle solution (sterile 0.1 M sodium citrate buffer, pH 4.5). The successful establishment of T1DM was confirmed by determining the fasting blood glucose level with Accu-Chek test strips (Roche Diagnostics, Indianapolis, IN, U.S.A.) 3 days after the STZ injection. All the STZ-injected mice presented with high levels of blood glucose (>16.7 mmol/L). No mouse died through the study period or required insulin supplementation to offset extreme weight loss, and at the study end, all STZ-injected mice remained hyperglycemic (fasting blood glucose > 16.7 mmol/L).