ABSTRACT: 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
The 18-amino-acid cationic, tryptophan-rich ApoEdpL-W peptide derived from human ApoE apolipoprotein was shown to have antifungal activity against pathogenic yeasts of the Candida genus (except C. glabrata). ApoEdpL-W was active against planktonic cells and early-stage biofilms but less active against mature biofilms, possibly because of its affinity for extracellular matrix beta-glucans. Moreover, ApoEdpL-W absorbed to medically relevant materials partially prevented the formation of biofilms o ...[more]
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:Invasion of host tissue by the human fungal pathogen, Candida albicans is an important step during many forms of candidosis. However, not all C. albicans strains possess the same invasive and virulence properties. It is known for example that the two clinical isolates SC5314 and ATCC10231 differ in their ability to invade into host tissue and to cause infections. Strain SC5314 is invasive whereas strain ATCC10231 is non-invasive and strongly attenuated in virulence as compared to SC5314. In this study we compare the in vitro transcriptional profiles and the genotypic profiles of these two widely used laboratory strains in order to determine the principal biological and genetic properties which may govern the different potential for invasiveness and virulence. Keywords: transcriptional profiling, comparative genomic hybridisation, invasive vs. non-invasive C. albicans strain Genomic DNA from C. albicans strains SC5314 and ATCC10231 hybridisations were done in duplicate including one dye swap. Total RNA from C. albicans strains SC5314 and ATCC10231 strains were compared in triplicate including one biological replicate and one dye swap.
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. 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: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: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:Azole resistance and varying degrees of cross-resistance to other members of the azole family in clinical isolates have been documented, which has necessitated additional and prolonged use of the antifungal agents available. 2-Amino-Nonyl-6-Methoxyl-Tetralin Muriate (10b), a novel chemical structural aminotetralin derivate, is synthesized as an antifungal agent and exibited strong antifungal activity. To further investigated the action mechanism, we used microarray analysis to investigate the genes expression profiles of C. albicans cells treated or untreated with 10b and found 957 genes were differentially expressed. Of them,457 showed a decrease in expression and 500 showed an increase in expression. 33 down-regulated genes were involved in glycolysis (e.g., PFK1, CDC19 and HXK2), fermentation (e.g., PDC11, ALD5 and ADH1) and respiratory electron transport chain (e.g., CBP3, COR1 and QCR8). 30 differentially expressed genes were found to relate to biofilm formation, filamentous or hyphal growth. It was noticed that striking up-regulation of SFL1 and marked down-regulation of YWP1 directly related to prevent C. albicans from changing its morphology from the yeast form to the hyphal. Two genes related to specifically hydrolyzing beta-1, 3 glucan (e.g., XOG1) and chitin (e.g., CHT1) were significantly increased. 40 overexpressed genes and 15 down-regulated genes were related to the lipid metabolic process. Of them, Eight were directly linked to ergosterol biosynthesis, including ERG2, ERG6 and ERG11. 99 genes related to translation were down-regulated following exposure to 10b, which account for 21.66% in down-regulated genes. This suggested that translation might be lower in SC5314 cells exposed to 10b than in control. Total RNA from the control SC5314 cells and 10b-treated SC5314 cells were used to generate target cDNA, and then hybridized to 8k Candida albicans Genome Array Genechips, representing about 7925 characterized Candida albicans genes. Two independent experiments were conducted. Reference strain was control SC5314 cells and test strain was SC5314 cells treated with 10b.
Project description:Candida albicans is an opportunistic pathogenic yeast that is commensally found in variety of host niches. Rhb1 serves as an positive regulator of Tor1 kinase which is the central component of the TOR signaling in controlling several virulence factors. Here, we used microarray to determined changes in transcript profile while the cell lacks the RHB1 gene. 10 array generated by six batchs of individual experiment (from cell isolation to culture); each included one flask of wild type culture (SC5314) and one flask of rhb1∆ (CCT-D1)mutant culture.
Project description:We perform microarray analysis of HUVECs upon stimulation with virulent wildtype C. albicans strain SC5314 or its efg1/efg1 cph1/cph1 hyphal-deficient derivative strain CAN34 to compare the gene expression profiles elicited from HUVECs in response to these strains. In addition, these responses are compared to that of TNF-alpha induced responses to determine which responses are Candida-specific. Experiment Overall Design: HUVECs are co-cultured for 3 or 8 hours in M199 medium alone or with CAN34, SC5314, or TNF-alpha. Total RNA is isolated, cRNA is synthesized and labeled, and labeled cRNA is hybridized onto Affymetrix chips. Each Candida and TNF sample is compared to its corresponding medium alone sample to determine fold changes in expression at each time point.