Fluconazole, supplied by InterChem, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Systematic discovery of multicomponent therapeutics
Journal: Proceedings of the National Academy of Sciences of the United States of America
Figure Legend Snippet: Multicomponent therapeutics that prevent proliferation of fluconazole-resistant C. albicans . All 435 possible two-component combinations of 30 compounds were tested in duplicate in 36-point dose matrices to identify synergistic combinations. C. albicans cells were treated with compounds in 384-well plates and the extent of inhibition of Alamar blue fluorescence was determined. ( A ) Overview of synergistic combinations in C. albicans Alamar blue proliferation assay. Yellow columns and rows indicate compounds that are commonly used as antifungal agents. Purple squares indicate combinations for which neither compound is used as an antifungal agent on its own and red indicates combinations for which one, but not both, of the compounds is used as an antifungal agent on its own. ( B ) A sample combination dose matrix, showing the combined effect of pentamidine and phenazopyridine at six concentrations (including a zero concentration point) each. The experimentally measured inhibition of Alamar blue signal is shown for each pair of combinations. The color of the squares also indicates the level of Alamar blue inhibition. ( C ) The calculated excess inhibition over the predicted Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( D ) The calculated excess inhibition over the HSA.
Techniques Used: Inhibition, Fluorescence, Proliferation Assay, Concentration Assay
Figure Legend Snippet: The combination of fluconazole and phenazopyridine selectively inhibits proliferation of fluconazole-resistant C. albicans .( A ) The percent inhibition of fluconazole-resistant C. albicans proliferation is shown for the indicated concentrations of fluconazole and phenazopyridine, determined by using an Alamar blue proliferation assay. The average of three measurements is shown. ( B ) The calculated excess inhibition over the Bliss additivism model. The predicted Bliss additive effect (see text) was subtracted from the experimentally observed inhibition at each pair of concentrations. ( C ) The calculated excess inhibition over the HSA. ( D ) Percent inhibition of fluconazole-resistant C. albicans proliferation at concentrations showing optimal synergy [250 μM fluconazole (flu) and/or 20 μM phenazolepyridine (PAP)]. ( E ) Fungicidal activity, determined by using a cfu assay. Fluconazole-resistant C. albicans were treated with 20 μM PAP and/or 250 μM flu, or 4 μM amphotericin B (amp B) as a fungicidal positive control, and in each case an equal number of yeast particles were plated in the absence of any compound. The number of colonies that grew after each treatment regime is indicated. ( F ) The combination of PAP and flu prevents dye efflux. Fluconazoleresistant C. albicans cells were treated with 20 μM PAP and/or 250 μM flu and the effect on efflux of rhodamine G was determined by using fluorescence microscopy. Phase-contrast images of the yeast cells are shown in each case to confirm that a large number of cells were present in each case, but that dye efflux was only effectively inhibited when both PAP and flu were present. For all numerical figures shown, the average of three measurements is represented. Error bars, 1 SD.
Techniques Used: Inhibition, Proliferation Assay, Activity Assay, Colony-forming Unit Assay, Positive Control, Fluorescence, Microscopy