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Metabolomic-based investigation on the effects of antifungal agents in Candida albicans

ABSTRACT: Our aim is to determine the metabolic effects of increasing doses of an antifungal agent on C. albicans metabolism (untargeted, steady state metabolomics). We will culture in vitro Candida cells to the mid-logarithmic growth in liquid media (RPMI-1640) at 37°C and then inoculate biological replicates (1ml) onto 22mm nitrocellulose filters under vocuum filtration in sterile conditions. Subsequently, isolates will be cultivated to midlogarithmic phase of growth on the same agar (RPMI-1640) to which the antifungal agent has been added at a range of concentrations to achieve doses equivalent to 0 MIC (no drug), 0.0625 MIC, 0.125 MIC, 0.25 MIC, 0.5 MIC and 1.0 MIC at 37°C. At mid-logarithmic phase of growth (12h) replicates will be metabolically quenched by immersion into a solvent mixture of 40% acetonitrile: 40% methanol: 20% water precooled at -40°C. The resulting quenched isolate/solvent mixtrue will be mechanically lysed by bead-beating with 0.1mm Zirconia beads in a tissue homogenizer and then centrifuged to seperate out cell wall components. Supernatants will be removed and stored at -80°C until they will be sent to SECIM facility.

ORGANISM(S): Candida Albicans

TISSUE(S): Yeast Cells

SUBMITTER: Aspasia Katragkou

PROVIDER: ST000243 | MetabolomicsWorkbench | Tue Feb 10 00:00:00 GMT 2015

REPOSITORIES: MetabolomicsWorkbench

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