Evaluating Antifungal Activities of C12 and C14

Representative time-kill studies were performed to investigate the activity of C₁₂ and C₁₄ in the presence or absence of POS against one azole-resistant strain, C. albicans ATCC 64124 (B). These assays were performed in 15 mL culture tubes using RPMI 1640 medium as previously described25 (link). Different sets of cell suspensions were prepared with C₁₂ (8 μg/mL), C₁₄ (4 μg/mL), and POS alone (10 μg/mL), or combinations of C₁₂ (2 μg/mL) plus POS (2.5 μg/mL) or C₁₄ (2 μg/mL) plus POS (1.25 and 2.5 μg/mL), or growth control (no drug) and sterility control (no cells and no drug). The final inoculum size of yeast cells used was 10⁵ cfu/mL as confirmed by colony count. The cell suspensions were then incubated at 35 °C with constant shaking (200 rpm). Aliquot of 100 μL from each tubes were removed at 0, 3, 6, 9, 12, and 24 h, and serially diluted in sterile ddH₂O. 50 μL of each dilution was plated onto potato dextrose agar (PDA) and then incubated at 35 °C. Colony counts were determined after 48 h of incubation. The experiments were performed in duplicate.