Insulin sensitivity was measured by performing an insulin tolerance test (ITT), and the rate constant for plasma glucose disappearance (Kitt) was calculated using a previously described method [5 (link), 27 (link)]. Recombinant human insulin (Novolin R, Novo Nordisk, Montes Claros, MG, Brazil) was administered at a dose of 2.0 U/kg, i.p. Blood samples were collected from the tail vein before (time 0) and 5, 10, 15, 20, 30 and 60 min after the insulin injection, and blood glucose levels were measured using standard test strips (One Touch UltraMini/Johnson & Johnson, Milpitas, CA, USA) [5 (link)]. The ITT is expressed as Kitt (%/min), i.e., the percent decrease in the plasma glucose concentration per minute [28 (link)].
One touch ultramini
Manufactured by Johnson & Johnson
Sourced in United States
The One Touch UltraMini is a handheld blood glucose monitoring device manufactured by Johnson & Johnson. It is designed to measure the level of glucose in a small sample of blood, providing users with a quick and convenient way to monitor their blood sugar levels.
Automatically generated - may contain errors
7 protocols using one touch ultramini
1
Insulin Sensitivity Measurement by ITT
2
Insulin Tolerance Test in Rats
3
Glucose Tolerance and Insulin Sensitivity Assay
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IP-GTT was performed after a 12-h fast as depicted in Fig. 1 . A 10 % glucose solution was injected i.p. (1 mg/g BW) and blood samples were obtained from the tail vein at 0, 15, 30, 60, 90, and 120 min after glucose administration as in [29 (link)]. Mice were tail bled at 0 wks (before treatment intervention), at 5 wks and at 10 wks (end of the study). Blood glucose was determined using a glucometer (Optium EZ, Abbot Laboratories and One Touch Ultra Mini, Johnson and Johnson) via test strips. Limit of detection of Optium EZ is 500 mg/dL (used in Study 1) and of One Touch Ultra Mini is 600 mg/dL (used in Study 1 and 2).![]()
Schematic drawing of in vivo experimental procedures. In study 1 and study 2, the mice (5 wks old) were randomly assigned to the various experimental treatments that were administered daily for 10 wks. An IP-GTT was performed at 0, 5 and 10 wks of the study, while an IP-ITT was performed at 10 wks. In each intervention blood was collected at 0 and 15 min post injection to detect insulin plasma levels
4
Oral Glucose Tolerance Test Protocol
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For the oral glucose tolerance test (OGTT), animals were fasted for 4 h (morning fast, from 8:00 a.m. until 12:00 p.m.), and blood samples were obtained from the tail vein. Animals were administered 2 g/kg bw of glucose by oral gavage, and blood samples were taken at 0, 15, 30, 45, 60, 90, and 120 min. The blood glucose was measured using blood glucose test strips (OneTouch® mini) and a glucometer (OneTouch® ultra mini-Johnson & Johnson). The AUC values were considered as glucose tolerance indexes.
5
Alloxan-Induced Diabetes Model Study
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C57/BL6, PAR2KO, and Glu-mT/mG mice were injected intravenously with 110 mg/kg alloxan (Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBS) at day 0. Mice with a blood glucose level >400 mg/dl (OneTouch Ultra Mini, Johnson & Johnson, Milpitas, CA, USA) were randomized. One group was injected with 0.1 mM/kg 2fLI (Santa Cruz Biotechnology, Santa Cruz, CA, USA) six IP injections on days 2, 4, and 6). Mice (alloxan +/−2fLI) were killed on days 2, 6, 9, 16, and 23:
Number of mice:
Day 2 – alloxan+vehicle, n=4 C57/BL6 and n=4 PAR2KO.
Day 6 – alloxan -2fLI, n=6 and n=3 PAR2KO; alloxan+2fLI, n=6 and n=3 PAR2KO.
Day 9 – alloxan -2fLI, n=10, n=4 PAR2KO, and n=3 Glu-mT/mG; alloxan+2fLI, n=10, n=4 PAR2KO, and n=3 Glu-mT/mG.
Day 16 – alloxan +2fLI, n=7, n=4 PAR2KO; alloxan -2fLI, n=7 and n=4 PAR2KO.
Day 23 – alloxan +2fLI, n=4, and n=3 Glu-mT/mG; alloxan -2fLI, n=4, and n=3 Glu-mT/mG.
Note that three C57/BL6 mice from each group at days 9 and 16 had BrdU (1 mg/ml, Sigma-Aldrich) added to the drinking water and that three C57/BL6 mice from each group at days 6 and 9 were taken for partial islet isolation (see below).
Number of mice:
Day 2 – alloxan+vehicle, n=4 C57/BL6 and n=4 PAR2KO.
Day 6 – alloxan -2fLI, n=6 and n=3 PAR2KO; alloxan+2fLI, n=6 and n=3 PAR2KO.
Day 9 – alloxan -2fLI, n=10, n=4 PAR2KO, and n=3 Glu-mT/mG; alloxan+2fLI, n=10, n=4 PAR2KO, and n=3 Glu-mT/mG.
Day 16 – alloxan +2fLI, n=7, n=4 PAR2KO; alloxan -2fLI, n=7 and n=4 PAR2KO.
Day 23 – alloxan +2fLI, n=4, and n=3 Glu-mT/mG; alloxan -2fLI, n=4, and n=3 Glu-mT/mG.
Note that three C57/BL6 mice from each group at days 9 and 16 had BrdU (1 mg/ml, Sigma-Aldrich) added to the drinking water and that three C57/BL6 mice from each group at days 6 and 9 were taken for partial islet isolation (see below).
6
Glucose Tolerance Test in Mice
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Multiparous dams fed their respective diets for at least 32 weeks underwent a fasting glucose tolerance test. Mice were fasted for 6 h, baseline glucose readings obtained and then they were given an IP injection of d ‐glucose (2 mg/g body weight). Glucose readings were taken 15, 30, 60, and 120 min after injection. Glucose was measured using a ONETOUCH Ultra Mini glucometer (Johnson & Johnson, New Brunswick, NJ) with commercially available UniStrip1 test strips (UniStrip Technologies, Charlotte, NC).
7
Alloxan-Induced Diabetes and Caerulein Treatment in Mice
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Mice were injected intravenously with 110 mg/kg alloxan (Sigma-Aldrich, St. Louis, MO, USA) in PBS (phosphate-buffered saline) at day 1. Mice with a blood glucose level greater than 500 mg/dl (OneTouch Ultra Mini, Johnson & Johnson, Milpitas, CA, USA) were randomized into two groups. One group was injected with caerulein (Sigma-Aldrich; six IP injections on days 1, 3 and 5). Lineage tracing studies were done with a caerulein dose of 7 mg/kg and the time course study with C57BL/6 mice used a dose of 10 mg/kg in PBS. Mice (alloxan alone and alloxan plus caerulein) were killed on days:
Day 13- Three C57BL/6 alloxan plus caerulein mice, three C57BL/6 alloxan alone mice.
Day 20- Three lineage tracing alloxan plus caerulein mice, three lineage tracing alloxan alone mice, seven C57BL/6 alloxan plus caerulein mice, seven C57BL/6 alloxan alone mice. Of the seven C57BL/6 alloxan plus caerulein mice and seven alloxan alone mice, one group from each (n=3) had BrdU (1 mg/ml, Sigma-Aldrich) added to the drinking water, whereas the second (n=4) had no BrdU added.
Day 27- Three C57BL/6 alloxan plus caerulein mice, three C57BL/6 alloxan alone mice.
Day 38- Three lineage tracing alloxan plus caerulein mice, three lineage tracing alloxan alone mice, four C57BL/6 alloxan plus caerulein and four C57BL/6 alloxan alone.
Day 13- Three C57BL/6 alloxan plus caerulein mice, three C57BL/6 alloxan alone mice.
Day 20- Three lineage tracing alloxan plus caerulein mice, three lineage tracing alloxan alone mice, seven C57BL/6 alloxan plus caerulein mice, seven C57BL/6 alloxan alone mice. Of the seven C57BL/6 alloxan plus caerulein mice and seven alloxan alone mice, one group from each (n=3) had BrdU (1 mg/ml, Sigma-Aldrich) added to the drinking water, whereas the second (n=4) had no BrdU added.
Day 27- Three C57BL/6 alloxan plus caerulein mice, three C57BL/6 alloxan alone mice.
Day 38- Three lineage tracing alloxan plus caerulein mice, three lineage tracing alloxan alone mice, four C57BL/6 alloxan plus caerulein and four C57BL/6 alloxan alone.
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