Amiodarone induces stress responses and calcium flux mediated by the cell wall in Saccharomyces cerevisiae.

We used a proteomic approach to study effects of amiodarone on cells of the yeast Saccharomyces cerevisiae. Amiodarone has been shown to have antifungal activity in vitro and causes a massive increase in cytoplasmic calcium levels ([Ca2+]cyt). Proteomic analysis of cells exposed to amiodarone show that this drug elicits stress responses and points to involvement of proteins associated with the cell wall. We tested several of those proteins for involvement in the Ca2+ flux. In particular, the amiodarone-induced Ca2+ flux was decreased in bgl2Delta cells, which have altered levels of beta-glucan and chitin. The involvement of the cell wall in the Ca2+ flux induced by amiodarone treatment was tested by addition of yeast cell-wall components. While mannan inhibited the rise in [Ca2+]cyt, beta-glucan potentiated the Ca2+ flux by 4.5-fold, providing evidence that the cell wall is directly involved in controlling this Ca2+ flux. This conclusion is corroborated by the inhibition of the Ca2+ flux by calcofluor, which is known to bind to cell-wall chitin and inhibit cell growth. Zymolyase treatment altered the kinetics of amiodarone-induced calcium flux and uncoupled the inhibitory effect of calcofluor. These effects demonstrate that the cell-wall beta-glucan regulates calcium flux elicited by amiodarone.

Enzyme immunoassay and immunoblotting analysis of Helicobacter pylori infection in Turkish asymptomatic subjects.

Approximately half of the world population is infected with Helicobacter pylori, particularly in developing countries. The aims of the study were to detect H. pylori infection in asymptomatic Turkish subjects, correlate the infection with the associated risk factors, and to evaluate the cytotoxin-associated gene (CagA) status and other H. pylori antigens. Three hundred nine asymptomatic subjects (124 female) 1-82 years of age (average: 31 years) were serologically tested by enzyme immunoassay and immunoblotting. The enzyme immunoassay detected IgG anti-H. pylori antibodies in sera of 216 (70%) out of 309 subjects, 132 (61%) male. Infection rates of 42% in subjects <10 years of age, 55% in 10-19 years, 66% in 20-29 years, 78% in 30-39 years, 79% in 40-49 years, 91% in 50-59 years, 100% in 60-69 years, and 80% in those >70 years of age were detected. Subjects >45 years of age had significantly higher antibody responses, odds ratio = 0.16 (95% confidence interval: 0.07-0.37), than those <45 years. H. pylori infection was significantly higher in married subjects, odds ratio = 0.38 (95% confidence interval: 0.20-0.73), and those with low socioeconomic status. No correlation between gender, education, smoking, and nonsteroidal anti-inflammatory drug intake and infection was detected. Immunoblots revealed antibodies to CagA in 58 (83%) of 70 samples tested. H. pylori infection is prevalent in the asymptomatic Turkish subjects. Marital and socioeconomic status was significantly associated with the acquisition of H. pylori. Antibodies to CagA antigen were highly prevalent in these subjects.