Project description:We examined the antifungal activity of artemisinin against Aspergillus fumigatus (A. fumigatus), a pathogenic filamentous fungus responsible for allergic and invasive aspergillosis in humans and analyzed transcript profiles of the fungus on exposure to Artemisinin. A. fumigatus spores were cultured for 48 h and then treated with artemisinin (at MIC50 concentration) or solvent control (DMSO) for 3 h to study its transcriptomic profiles.
Project description:Genomic DNA from five strains, Aspergillus fumigatus Af71, Aspergillus fumigatus Af294, Aspergillus clavatus, Neosartorya fenneliae, and Neosartorya fischeri, were co-hybridized with that of Aspergillus fumigatus Af293 and compared.
Project description:Aspergillus fumigatus is the most important pulmonary fungal pathogen and is able to cause several diseases collectively called aspergillosis. Conidia is the most important infection structure making the initial contact with the human host. Here, we used a phylogenomic approach comparing proteins present in the A. fumigatus conidial surface, two closely related non-pathogenic species A. fischeri and A. oerlinghausenensis, and the far-related pathogenic A. lentulus. We were able to identify 62 proteins specifically expressed on A. fumigatus conidial surface. We deleted 42 of the encoding-genes and observed that many of them have altered susceptibility to macrophage killing, penetration and damage to epithelial cells, and cytokine production. We demonstrated that one of these genes encoding a glycosylasparaginase is modulating IL-1β levels and is important for the infection in an immunocompetent murine model. Our results provide opportunities for characterizing A. fumigatus effectors important for evasion and modulation of the immune response.
Project description:Acyl-homoserine lactone (acyl-HSL) quorum sensing was first discovered in Vibrio fischeri where it serves as a key control element of the seven-gene luminescence (lux) operon. Since this initial discovery, other bacteria have been shown to control hundreds of genes by acyl-HSL quorum sensing. Until recently, it has been difficult to examine the global nature of quorum sensing in V. fischeri. However, the complete genome sequence of V. fischeri is now available and this has enabled us to use transcriptomics to identify quorum-sensing regulated genes and to study the quorum-controlled regulon of this bacterium. In this study, we used DNA microarray technology to identify over two-dozen V. fischeri genes regulated by the quorum sensing signal N-3-oxohexanoyl-L-homoserine lactone (3OC6-HSL). Keywords: Comparison of transcriptome profiles
Project description:Aspergillosis covers a range of infections cause by Aspergillus species, and in many cases can be life threatening. Individuals with weakened immune systems are particularly at risk. Dendritic cells were derived from patients with allergic brochopulmonary aspergillosis (ABPA), chronic cavitary pulmonary aspergillosis (CCPA), and asthma, as well as from healthy donors. Each sample was split in two, and one sample from each pair was cultured with Aspergillus fumigatus. RNA-sequencing was used to assess transcriptional changes in the human cells in response to pathogen challenge. Many genes known to be important in immunity were found to exhibit differential expression after fungal challenge between healthy and diseased individuals, including chemokines and C-type lectins.
Project description:The marine bacterium Vibrio fischeri requires flagellar motility to undergo symbiotic initiation with its host, the Hawaiian bobtail squid Euprymna scolopes. We sought to identify the genes activated by the sigma54-dependent flagellar master regulator, FlrA, in V. fischeri, thereby determining the flagellar regulon in this model symbiont.
Project description:COVID-19-associated pulmonary aspergillosis (CAPA) is a frequent, severe superinfection with the fungus Aspergillus affecting critically ill COVID-19 patients. The pathophysiology of this infection is still largely unknown. We performed an explorative study by performing single-cell RNA sequencing (scRNA-seq) on 43 bronchoalveolar lavage (BAL) fluid samples from 39 mechanically ventilated COVID-19 patients.
Project description:The marine bacterium Vibrio fischeri requires flagellar motility to undergo symbiotic initiation with its host, the Hawaiian bobtail squid Euprymna scolopes. We sought to identify the genes activated by the sigma54-dependent flagellar master regulator, FlrA, in V. fischeri, thereby determining the flagellar regulon in this model symbiont. We performed microarray analysis on wild-type Vibrio fischeri ES114 and a flrA deletion mutant, DM159, grown to mid-log phase in seawater tryptone, a condition in which cells are highly motile (two biological replicates per condition).
Project description:Aspergillus fumigatus is a filamentous fungus capable to cause an important disease known as invasive aspergillosis. Challenge different cell lines is a crucial strategy to undercover new virulence factors involved in the infection process. In this research, RAW 264.7 macrophages and A549 lung epithelial cells were challenge using A. fumigatus conidia in a MOI of 10 and 5 respectively. When these conidia reach 30% of germination, the total RNA was isolated (A. fumigatus alone (Control), A. fumigatus vs RAW 264.7, and A. fumigatus vs A549) and purified using the RNeasy Plant Mini Kit (Qiagen). The AWAFUGE (Agilent Whole A. fumigatus Genome Expression 44K v.1) hybridization was carried out following previously publish protocols (A-MEXP-2352 and E-MTAB-5314).
