Freestyle lite blood glucose monitoring system

Body weights were measured and recorded weekly throughout the course of the study. The implanted insulin pellets issued ~ 2 doses per day at a constant release rate, and non-fasted BG readings were taken twice weekly at 9 am by collecting a small droplet of blood (~ 50 μL) from the saphenous vein throughout the duration of the study. BG values were analyzed via a Freestyle Lite Blood Glucose Monitoring System (Abbott Diabetes Care, INC. Conyers, GA, USA) and reported in millimoles per litre (mmol/L).

All animal work has been approved by IACUC committee of Weill Cornell Medical College. CD-1 adult male mice were treated with different doses of propargite or vehicle (corn oil) through intraperitoneal injection for 5 days. Fasting blood glucose level were monitored every day using Abbott FreeStyle Lite blood glucose monitoring system. Then, CD-1 adult male mice were treated with 12 mg/kg propargite or vehicle (corn oil) through intraperitoneal injection for 5 days. Fasting blood was collected via tail nicking following 14–16 h of fasting with ad libitum access to drinking water. Serum mouse insulin levels were assessed by ELISA (80-INSMR-CH01, Alpco). Mice were then killed. The pancreases were embedded in OCT. The tissue was then sectioned at 5 μm for immunohistochemistry staining analysis of DNA damage marker γ-H2A.X and β-cell marker INS.

After blood glucose levels had risen above ~20 mM (~2 days after tamoxifen injection), animals were immediately implanted subcutaneously with either a (1) placebo pellet, (2) 60-day slow-release glibenclamide pellet (17 mg kg⁻¹ per day) (Innovative Research of America), or (3) two to three 30-day (~0.1 U per 24 h) insulin pellets (body weight <30 g, 2 pellets; >30 g, 3 pellets) (LinShin Canada Inc.) under 2% isoflurane anaesthesia. Placebo pellets contained vehicle but no drug. In separate experiments, Kir6.2-V59M expression was induced, mice left diabetic for 4 weeks (blood glucose >20 mM), then treated with glibenclamide (2-3 pellets; 34–95 mg kg⁻¹ per day), and studied 4 weeks later.
Free-fed plasma glucose levels were measured daily (at 1400, h; FreeStyle Lite Blood Glucose Monitoring System, Abbott). Insulin and glucagon were measured in plasma or whole islets by radioimmunoassay (Millipore and Euro Diagnostica, respectively).

Reagents, molecular weight markers, and nitrocellulose membranes for all western blotting experiments were purchased from BioRad Laboratories. All BCA assay reagents were purchased from Fisher Scientific. The following antibodies were provided as gifts: FAT/CD36 (M025) from Dr. N.N. Tandon (Otsuka Maryland Medical Laboratories), FABPpm from J. Calles‐Escandon (Wake Forest University), and MCT‐1 from Dr. H Hatta (Tokyo University). Insulin (Humulin rDNA origin) and FreeStyle Lite blood glucose monitoring system (Abbott Diabetes Care Canada) were purchased from Shoppers Drug Mart. All other reagents are listed in Table 1.