Tryptophan 650 of human granulocyte colony-stimulating factor (G-CSF) receptor, implicated in the activation of JAK2, is also required for G- CSF-mediated activation of signaling complexes of the p21ras route

Granulocyte colony-stimulating factor (G-CSF) induces rapid phosphorylation of JAK kinases as well as activation of the p21ras route through interaction with its specific receptor (G-CSF-R). The cytoplasmic membrane-proximal region of G-CSF-R (amino acids 631 to 684) is necessary for proliferation induction and activation of JAK2. In contrast, activation of Shc and Syp, signaling molecules implicated in the p21ras signaling route, depends on the phosphorylation of tyrosine residues located in the membrane-distal region (amino acids 685 to 813) of G-CSF-R. We investigated whether G-CSF-induced activation of signaling complexes of the p21ras route depends on the function of the membrane-proximal cytoplasmic region of G-CSF-R. A G- CSF-R mutant was constructed in which tryptophan 650 was replaced by arginine and expressed in BAF3 cells (BAF/W650R). In contrast to BAF3 cell transfectants expressing wild-type G-CSF-R, BAF/W650-R cells did not proliferate and did not show activation of JAK2, STAT1, or STAT3 in response to G-CSF. Immunoprecipitations with anti-Shc and anti-Grb2 antisera showed that mutant W650R also failed to activate Syp and Shc. These data indicate that the membrane-proximal cytoplasmic domain of G- CSF-R is not only crucial for proliferative signaling and activation of JAK2 and STATs, but is also required for activation of the p21ras route, which occurs via the membrane-distal region of G-CSF-R.     

Effect of air-abrasion pre-treatment with bioactive glass 45S5 on enamel surface loss after erosion/abrasion challenge

ObjectivesThe aim of this in vitro study was to evaluate the effect of air-abrasion surface pre-treatment with bioactive glass (BAG) 45S5 on enamel surface loss after erosion/abrasion challenge.MethodsTwenty-four sound bovine incisors were used for this study. Four experimental groups (n = 12) were assigned as follows: Group 1 was the negative control group, Group 2 specimens were treated with a SnF₂ gel (positive control group), Group 3 specimens were air-abraded with BAG 45S5 (ProSylc) and Group 4 received both treatments. The specimens were submitted to erosion/abrasion challenge using a common soft drink. Enamel surface loss was evaluated using an optical profilometer. Additionally, surface roughness (VSI method) and surface microhardness (Vickers method) changes were evaluated, as well as SEM and EDS analyses were performed on enamel surface. The data were statistically analyzed using one-way ANOVA and Tukey’s post-hoc test at a level of significance a = 0.05.ResultsSurface pre-treatment with BAG 45S5 reduced surface loss in comparison with negative control group (p < 0.001), which exhibited the highest surface loss of the experimental groups (p < 0.05). The positive control group (SnF₂ treatment) and Group 4 specimens, which received both air-abrasion pre-treatment and SnF₂ treatment, presented the lowest surface loss (p < 0.05), but did not show significant differences to each other (p = 0.65).SignificanceAir-abrasion pre-treatment with BAG 45S5 may be beneficial as an in-office preventive method for the limitation of enamel erosive wear induced by excessive consumption of soft drinks. The clinical significance of the results regarding this preventive method should be confirmed by clinical studies.