Accu check performa

Cholesterol concentration in all lipoprotein fractions was measured by enzymatic colorimetric kit (Roche Diagnostic, Brazil). HDL-c was determined at the final period only by the ratio: HDL cholesterol area/total cholesterol area. Total plasma cholesterol and triglycerides were determined by enzymatic techniques (Labtest, Brazil) and glucose by Accu Check Performa glucometer (Roche, Brazil). Lipoprotein profile was determined by fast protein liquid chromatography (FPLC) gel filtration on two Superose 6 columns.

Six healthy controls and six diabetic New Zealand white rabbits were used in this study. Mini-implants were placed 4 weeks after diabetic induction. Those mini-implants were removed in 4 control rabbits and 4 diabetic rabbit following 4 weeks of healing. Micro-CT and histological analysis were performed in four rabbits. The experiment protocol was approved by the Institutional Animal Care and Use Committee. (CUMC-2010-0093-01).
For the alloxan injection, rabbits were lightly anesthetized with ketamine hydrochloride 30 mg/kg and xylazine 3 mg/kg (IM). Alloxan monohydrate (Sigma Aldrich Chemical, Saint Louis, MO, USA) was dissolved in sterile normal saline to achieve concentration of 5% (W/V), and 100 mg/kg was immediately administered intravenously via marginal ear vein for 2 minutes. 4, 8, and 12 hours after alloxan injection, 10 mL of glucose (5% W/V) was administered subcutaneously [15 ]. Four days after the injection, blood samples were collected from aural vein and blood glucose level was monitored using ACCU-CHECK Performa (Roche Diagnostics, Mannheim, Germany). If glucose level was over 200 mg/dL, diabetes was diagnosed [16 (link)]. Blood glucose level and weight of rabbits were also monitored weekly to check the diabetic state.

Following the dietary/drink intervention and body composition scan, the mice were anesthetized with gaseous isoflurane and blood samples were collected via cardiac puncture. Whole blood was collected and was used to measure glucose by using a point-of-care Accu-Check Performa (Roche, Basel, Switzerland). Blood levels of glycated haemoglobin (HbA1c) were also analysed immediately after cardiac puncture by using DCA Vantage Analyzer (Siemens, Munich, Germany) as described previously [32 (link)]. Subsequently, blood samples were allowed to clot at room temperature for 30 min and were centrifuged for 10 min at 4000 rpm. Thereafter, the supernatant serum was aliquoted and stored at −80 °C for further analysis.

Glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed in mice before and after a 15-week HFD feeding. For the GTT, mice were fasted for 16 h with free access to water, and then orally gavaged with 1 g/kg body weight of glucose. Blood glucose level was assessed with the Accu-Check Performa (Roche Applied Science, Mannheim, Germany) at 0, 15, 30, 60, 90, and 120 min. For the ITT, mice were fasted for 6 h with free access to water, and then i.p. injected with 1 U/kg of recombinant human insulin (Novolin 30R; Novo Nordisk, Bagsvaerd, Denmark). Blood glucose level was measured at 0, 15, 30, 60, and 120 min after insulin injection.

All experiments were approved by the Animal Care and Use Committee at the Lebanese American University (LAU) and adhered to the Guide for the Care and Use of Laboratory Animals published by the US National Research Council committee.
Eighty‐nine adult male Wistar rats were used in this study. T1D was induced at 5 weeks of age in rats weighing between 80–130 g as previously described.³⁷ Briefly, 12 h after fasting, animals received one intraperitoneal injection of streptozotocin (STZ, Sigma‐Aldrich: 65 mg/kg in 0.1 M citrate buffer, pH = 4.5). Age‐matched control rats (CON) were injected with vehicle (0.1 M citrate buffer, pH = 4.5) via the same route. After 72 h, fasting glucose levels were measured in blood droplets withdrawn from the tails of animals, using Accu‐Check® Performa glucometer (Roche). Eighty‐five per cent of the rats injected with STZ exhibited fasting blood glucose (FBG) levels >200 mg/dL in addition to polyuria and polydipsia and were considered diabetic. All experiments were performed at 4, 8 and 12 weeks after STZ or vehicle injection (Figure S1).

After 5 months of diet, an intraperitoneal glucose tolerance test (IPGTT) was performed to evaluate glucose homeostasis. Animals were fasted for six hours, with free access to water during the entire experiment. Rats were weighed and the extremity of the tail was incised in order to obtain a small drop of blood, which was placed on the test strip of the blood nano-glucometer ACCU-CHECK Performa (Roche, Bale, Switzerland). The value obtained corresponds to the fasting blood glucose level (T0). Then a bolus of glucose was injected in the intraperitoneal cavity at the dose of 1 g/kg. Glycemia was measured 15, 30, 60, 90, 120 min after the injection of glucose.
Systolic and diastolic blood pressure were recorded non-invasively by the tail-cuff method (Bioseb, Chaville, France). The animals were lightly sedated with isoflurane (1%), the body temperature was maintained at 38.5 °C using a heating blanket, and blood pressure and heart rate were recorded (10 measurements). Averaged values for each rat were used for the subsequent statistical analysis.
At the time of sacrifice, amounts of subcutaneous and visceral adipose tissues were dissected and weighed. The sum of gonadal, mesenteric, retroperitoneal and perirenal fat was considered as visceral fat. Fat percent was determined using the following formula: ((subcutaneous + visceral adipose tissues) × 100)/total body weight.