But the compromised clearance of uninfected RBCs or erythroid response could not be excluded as a possibility

lony formation was limited. Using higher concentrations viable counts were 81% and 90% of the control. Therefore, again, at the microcolony level the fungicidal activity of these drugs was marginal. These trends were similar for other echinocandin sensitive isolates, i.e. strains ATCC204305, JBZ17 and JBZ32 of A. fumigatus and strain CWZ59 of A. terreus. In contrast, amphotericin B reduced viability,86% when used at the MICs of JBZ11, JBZ13 and JBZ32. Quantification of cell lysis by anidulafungin and caspofungin. Cell lysis was scored for a range of concentrations of caspofungin and anidulafungin after 14 h. Lysis was particularly common at intermediate concentrations around the MIC. In contrast, whilst higher concentrations of echinocandins were more effective at limiting MedChemExpress LY3039478 growth lysis was seen less often. Anidulafungin appeared somewhat more effective than caspofungin, with.50% cell lysis at the most effective concentrations. Fungal microcolonies growing on echinocandins appeared heterogeneous in their response. Subpopulations of lysed cells and intact ones coexisted within the same microcolony, ones that were derived, in most cases, from a single conidium. In both cases, a total count of microcolonies after 14 h suggested that neither drug reduced the number of microcolonies, despite having a significant impact on the number of intact hyphal tips within a microcolony. Caspofungin resistance increased the concentration of this drug required for optimal tip lysis commensurate ” with the increased MIC compared to sensitive strains. A series of control experiments were performed, using different staining or fixing and imaging methods, to check that the “6145492 observed tip lysis was not affected by sample preparation or by dye choice. The latter point was considered, in part, as calcofluor white is known to destabilise the cell wall in A. nidulans. Fun1/calcofluor white staining, propidium iodide/Syto9 staining, scanning electron microscopy with vapour or gel fixation all gave similar frequencies of lysis. Therefore it can be concluded that imaging or staining methods did not bias the results.Correlation between observed lysis and staining with Syto9 or propidium iodide for anidulafungin. Microcolonies of A. fumigatus cultured from heterogeneity was observed, most completely with propidium iodide. microcolonies stained Recovery of microcolonies of A. fumigatus from the effect of anidulafungin and caspofungin. PAO strips were moved from plates containing echinocandins to those lacking the drugs in order to look at the potential for recovery from echinocandins. Control experiments were first performed by moving uninoculated strips from plates containing echinocandins to drug free plates and then inoculating with A. fumigatus conidia. No inhibition of growth was seen, suggesting that the carry-over of drugs within the minimal volume of the PAO pores was not sufficient to influence the results. It was clear from assessment of the average microcolony diameter at different time points that after removal recovery was widespread throughout the population. Microcolonies cultured for 14 h on plates containing 0.125 mg/ml anidulafungin were stained with propidium iodide and Syto9 and compared with those cultured for 17 h on the same concentration or those shifted to zero anidulafungin after 14 h then grown for 3 h. In all cases lysed hyphal tips stained with propidium iodide. Furthermore, there was no evidence for extension of a lysed hyphal tip beyond the de