Tyrosine phosphatase and cytochrome P450 activity are critical in regulating store-operated calcium channels in human fibroblasts

Diverse signaling pathways have been proposed to regulate store-operated calcium entry (SOCE) in a wide variety of cell types. However, it still needs to be determined if all of these known pathways operate in a single cell type. In this study, we examined involvement of various signaling molecules in SOCE using human fibroblast cells (HSWP). Bradykinin (BK)-stimulated Ca2+ entry, previously shown to be via SOCE, is enhanced by the addition of vanadate, an inhibitor of tyrosine phosphatases. Furthermore, SOCE is regulated by cytochrome P-450, as demonstrated by the fact that the products of cytochrome P-450 activity (14,15 EET) stimulated SOCE while econazole, an inhibitor of cytochrome P450, suppressed BK-stimulated Ca2+ entry. In contrast, Ca2+ entry was unaffected by the guanylate cyclase inhibitor LY83583, or the membrane permeant cyclic GMP analog 8-bromo-cyclic GMP (8-Br-cGMP). Neither nitric oxide donors nor phorbol esters affected BK-stimulated Ca2+ entry. SOCE in HSWP cells is primarily regulated by tyrosine phosphorylation and the cytochrome P-450 pathway, but not by cyclic GMP, nitric oxide, or protein kinase C. Thus, multiple pathways do operate in a single cell type leading to the activation of Ca2+ entry and some of these signaling pathways are more prominently involved in regulating calcium entry in different cell types.

Disk diffusion susceptibility testing of dermatophytes with imidazoles.

In vitro susceptibility testing of 43 isolates of dermatophytes was carried out against imidazoles-ketoconazole, miconazole and econazole and griseofulvin by agar dilution and disk diffusion methods. Econazole was the most effective drug inhibiting all the isolates at a concentration of 0.1 microgram ml-1. The MIC 50s and MIC 90s for ketoconazole and miconazole were 1 and 2.5 mg ml-1 whereas the values for griseofulvin were 1 and 5 micrograms ml-1. Good correlation was seen between the MIC and sizes of zones of inhibition around the disks. Regression analysis was used to measure the degree of correlation between the MIC values and matched averaged zones of inhibition and the correlation coefficients for econazole, ketoconazole, miconazole and griseofulvin were -0.5554, -0.5886, -0.8558 and -0.8268 (p < 0.001) respectively.

In vitro evaluation of three topical antimycotics against ring-worm fungi singly and in combination.

Three antimycotic drugs, viz., Miconazole nitrate, Econazole nitrate and ciclopirox olamine were tested singly and in combination of miconazole nitrate and Econazole nitrate, Miconazole nitrate and Ciclopirox olamine, and Econazole nitrate and Ciclopirox olamine to evaluate in vitro efficacy against Trichophyton mentagrophytes and Macrosporum nanum. The best efficacy was shown by Ciclopirox olamine (MIC 0.78 microgram/ml) and a combination of Miconazole nitrate and Econazole nitrate (MIC 0.78 microgram/ml).