ABSTRACT: We found that the antibiotic colistin acts synergistically with antifungals of the echinocandin class (e.g. aminocandin) on C. albicans cells. In order to elucidate the mode of action of colistin in fungi we performed microarray analysis of samples treated with only aminocandin (0.00125µg/ml) or treated with aminocandin (0.00125µg/ml) and colistin (5µg/ml). We compared: (A). untreated cells to cells treated with aminocandin only; (B). cells treated with aminocandin to cells treated with aminocandin and colistin. By comparing those datasets it should be possible to identify genes differentially expressed in response to aminocandin and in response to both drugs. And subsequently to be able to interpret where in the cell colistin acts. (See related experiment in ArrayExpress: E-MEXP-3438)
Project description:We found that the antibiotic colistin acts synergistically with antifungals of the echinocandin class (e.g. aminocandin) on C. albicans cells. In order to elucidate the mode of action of colistin in fungi we performed microarray analysis of samples treated with only aminocandin (0.00125µg/ml) or treated with aminocandin (0.00125µg/ml) and colistin (5µg/ml). We compared: (A). untreated cells to cells treated with aminocandin only; (B). cells treated with aminocandin to cells treated with aminocandin and colistin (which is the focus of this experiment). By comparing those datasets it should be possible to identify genes differentially expressed in response to aminocandin and in response to both drugs. And subsequently to be able to interpret where in the cell colistin acts. (See related experiment in ArrayExpress: E-MEXP-3437 for comparison between untreated cells vs cells treated with aminocandin only.)
Project description:We performed a gene expression analysis of C. albicans SC5314 planktonic cells exposed to the antifungal peptide ApoEdpL-W. Exponentially-growing C. albicans SC5314 cells in SD at 30°C medium were exposed to 2.5 µM ApoEdpL-W and samples were collected after 10 and 30 min. for transcript profiling
Project description:Candida albicans wild type SC5314 and the eed1 delta mutant were used to infect reconstituted human oral epithelium (RHE) for 24h at 37°C and 5% CO2. Samples were taken 1h, 12h and 24h after infection. Total RNA was isolated, labeled with Cy5 and hybridised with a Cy3- labeled common reference.
Project description:An exponentially growing C. albicans SC5314 culture in SD medium at 30 C was split into two flasks, one exposed to MIC90 concentration of compound 2 (6.25 ug./ml-1)mannich ketones, the other to the same volume of water. Samples were collected after 30 and 60 min for transcript profiling.
Project description:3 replicates of WT and srbA knockout strains grown on GMM plates, harvested spores and inoculated 5x10^5 spores/ml in 300 ml of LGMM. Cultures incubated for 10h at 28C and then shifted for 2h to 37C (both at 200rpm) so they are just germinating at the start of the treatment. Cultures were shifted to hypoxia (37C, 200rpm) and incubated for 1hr. Samples were collected by pouring them over a filter paper in a Buechner funnel and snap froze the cells immediately in liquid nitrogen, and stored the cells at -80C and lyophilized all tissue at the same time after all samples were collected. RNA extraction was then done simultaneously with all time points.
Project description:An exponentially growing culture of strain R6 in AGCH at OD620nm=0.4 was either non-treated or treated with two LVX concentrations: 0.125 ug/ml LVX (0.5x MIC of R6) and 2.5 ug/ml LVX (10x MIC of R6). Samples were taken before treatment (0 min), at 15, 30 and 60 min in the non-treated culture, and at 5, 15 ,30 and 60 min in the LVX treated cultures
Project description:C. albicans wild type strain SC5314, the eed1 deletion mutant and an eed1 delta mutant overexpressing UME6 (eed1 + pTET-UME6) were grown on plastic (37°C, RPMI1640 medium, plastic surface, 5% CO2) for 12h. Total RNA was isolated using a phenol-chloroform protocol and labeled with Cy5. Cy5- labeled sample RNA was hybridised with Cy3- labeled common reference (SC5314, 37°C, exponential culture).
Project description:To assess the effect of carbon nanotubes substrated on dendritic cell (DCs) properties, DCs were generated from human monocytes of healthy donors as previously described (Aldinucci A et al, 2010). In particular, isolated monocytes were cultured for 6 days in medium supplemented with GM-CSF (1000U/ml) and IL-4 (1000U/ml), in presence or absence of multi-walled carbon nanotubes (MWCNT); then DCs were activated by 24 hours of incubation with LPS (1ug/ml). RNA extracted from floating and CNT adherent DCs were then used for transcriptional profilingthen used
Project description:2 million E. histolytica HM1:IMSS was treated with 0.5% DMSO for 3 h and 2 million E. histolytica HM1:IMSS was treated with 1 M-5M auranofin for 3 h