Glucose- and insulin-induced phosphorylation of the insulin receptor and its primary substrates IRS-1 and IRS-2 in rat pancreatic islets

The presence of tyrosine-phosphorylated proteins was studied in cultured rat pancreatic islets, Immunoblotting performed with total extracts of islets cultured in the presence of 1.8 or 5.6 mM glucose revealed at least three distinct tyrosine-phosphorylated bands (25 kDa, 95 kDa and 165-185 kDa). After 12 h incubation in medium containing 1.8 mM glucose, a pulse exposition to 11 or 22 mM glucose or to 10(-7) M insulin led to a substantial increase in the phosphorylation of all three bands, with no appearance of novel bands. Immunoprecipitation with specific antibodies demonstrated that the signal detected at 95 kDa corresponds to the beta subunit of the insulin receptor (IR) while the band at 165-185 kDa corresponds to the early substrates of the insulin receptor, IRS-1 and IRS-2. Immunoprecipitation with IRS-I or IRS-2 antisera detected their association with the lipid metabolizing enzyme phosphatidylinositol 3-kinase (PI 3-kinase), Thus, this is the first demonstration that elements involved in the insulin-signalling pathway of traditional target tissues are also present in pancreatic islets and are potentially involved in auto- and paracrine-signalling in this organ.

Root instrumentation with an erbium : yttrium-aluminum-garnet laser: Effect on the morphology of fibroblasts

Objective: the purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet laser instrumentation of root surfaces on the morphology of fibroblasts from continuous lineage. Method and materials: Dentinal slices with 4 mm(2) of surface area were obtained from teeth extracted for severe periodontal involvement. Specimens were assigned to one of three treatment groups: group 1, application of the laser with an energy level of 250 mJ at 103 pulses per second; group 2, application of the laser with an energy level of 80 mJ at 166 pulses per second; and group 3, similar to group 2, but with concomitant water irrigation of the device. The specimens were incubated in multiwell plates containing cell culture media. After 24 hours, the specimens were submitted to routine preparation for scanning electron microscopy. Three independent and blind examiners used photomicrographs to evaluate the morphology of the fibroblasts: 0 = without cells; 1 = flat cells; 2 = round cells; and 3 = combination of round and flat cells. Results: Statistical analysis indicated that there were significant differences among treatment groups and that group 3 was significantly different from groups 1 and 2. Conclusion: There was no difference between groups 1 and 2 in the morphology of fibroblasts. Laser instrumentation with concomitant irrigation impaired the adhesion of fibroblasts to dentinal surfaces.

Glucose reactivity with filling materials as a limitation for using the glucose leakage model

Aim To evaluate the reactivity of different endodontic materials and sealers with glucose and to asses the reliability of the glucose leakage model in measuring penetration of glucose through these materials.Methodology Ten uniform discs (radius 5 mm, thickness 2 mm) were made of each of the following materials: Portland cement, MTA (grey and white), sealer 26, calcium sulphate, calcium hydroxide [Ca(OH)(2)], AH26,Epiphany, Resilon, gutta-percha and dentine. After storing the discs for 1 week at 37 degrees C and humid conditions, they were immersed in 0.2 mg mL(-1) glucose solution in a test tube. The concentration of glucose was evaluated using an enzymatic reaction after 1 week. Statistical analysis was performed with the ANOVA and Dunnett tests at a significant level of P < 0.05.Results Portland cement, MTA, Ca(OH)(2) and sealer 26 reduced the concentration in the test tube of glucose significantly after 1 week (P < 0.05). Calcium sulphate reduced the concentration of glucose, but the difference in concentrations was not significant (P = 0.054).Conclusions Portland cement, MTA, Ca(OH)(2) and sealer 26 react with a 0.2 mg mL(-1) glucose solution. Therefore, these materials should not be evaluated for sealing ability with the glucose leakage model.