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The minimal effective concentration (MEC) was determined only for caspofungin, defined as the lowest drug concentration at which short and branched hyphae were observed [11]. Antifungal susceptibility testing was additionally performed with the commercial Sensititre YeastOne panel (Trek Diagnostic System Ltd, East Grinstead, UK) according to the manufacturer��s instructions. Colorimetric MEC was defined as the lowest concentration visibly reducing growth (despite presence of red or purple colour). We added the colorimetric Sensititre YeastOne assay for comparison of susceptibility testing because several routine microbiology laboratories are using this method rather than the CLSI-reference microbroth dilution. For microcalorimetric evaluation of growth characteristics, Sabouraud dextrose broth (SDB) (Oxoid CM0147; Basingstoke, Hampshire, UK) was used (3?mL medium and 1?mL air in the headspace of the ampoule). An inoculum of c. 2.5?��?104?conidia/mL Imatinib datasheet PD98059 was used, determined by microscopic enumeration using a Neubauer haematocytometer. Air-tightly sealed ampoules were introduced into the isothermal microcalorimeter (TAM III; TA Instruments, Newcastle, DE, USA) and measurements were performed at 37��C every 10?s. The detection threshold was determined at 5?��W to distinguish fungal heat production from the thermal background noise (i.e. growth media without moulds). The detection time (in h), heat-flow peak (in ��W), the time to peak (h) and total heat produced (in Joules, represented by the area under the heat-flow curve) at 24, 48 and 72?h were determined. For testing antifungal agents, two-fold dilutions of antifungal agents were added to SDB. The minimal heat inhibitory concentration (MHIC) of amphotericin B and triazoles was defined as the lowest antifungal concentration inhibiting 50% of the total heat produced by the growth control at 48?h. For echinocandins, the MHIC was defined as the lowest concentration reducing the heat-flow peak Carnitine palmitoyltransferase II by 50%. Susceptibility testing experiments were performed in duplicate. Data analysis was carried out with the manufacturer��s software (TAM Assistant; TA Instruments) and GraphPad Prism 5.0 (GraphPad Software, La Jolla, CA, USA). The thermokinetic characteristics of the Aspergillus strains tested in the absence of antifungals are presented in Table?S1. The median heat detection time was