Project description:This project has two goals. Firstly, to compare the gene expression profiles of Caco cells following exposure to Verocytotoxigenic E. coli0157:H7 (VTEC) isolates from food animals (bovine, ovine, porcine) and human in an effort to assess the invasive and toxigenic potential of isolates of different origin. All sources contain the common virulence and type 3 secretory system genes. Secondly, to compare the gene expression profiles of Caco-2 cells following exposure to VTEC isolates that contain (positive) or do not contain (negative) the genes of the type 3 secretory system (TTSS).
Project description:CF patients suffer from chronic and recurrent respiratory tract infections which eventually lead to lung failure followed by death. Pseudomonas aeruginosa is one of the major pathogens for CF patients and is the principal cause of mortality and morbidity in CF patients. Once it gets adapted, P. aeruginosa can persist for several decades in the respiratory tracts of CF patients, overcoming host defense mechanisms as well as intensive antibiotic therapies. P. aeruginosa CF strains isolated from different infection stage were selected for RNA extraction and hybridization on Affymetrix microarrays. Two batch of P. aeruginosa CF isolates are chosen : 1) isolates from a group of patients since 1973-2008 as described in ref (PMID: 21518885); 2) isolates from a group of newly infected children as described in ref (PMID: 20406284).
Project description:In 2014, enterovirus D68 (EV-D68), previously associated primarily with mild respiratory illness, caused a large outbreak of severe respiratory illness and, in rare instances, paralysis. We compared viral binding and replication of eight recent EV-D68 clinical isolates and the prototype Fermon strain from 1962 in cultured HeLa cells and differentiated human primary bronchial epithelial cells (BEC) to understand the possible reasons for the change in virus pathogenicity. We found no significant differences in binding or replication in HeLa cell cultures between the recent clinical isolates. However, in HeLa cells, Fermon had significantly greater (1.5-2 log) binding and virus progeny yields but a similar level of replication (~2-log increase in viral RNA from 2h to 24h post infection) compared to recent isolates. In differentiated BECs, Fermon and the recent EV-D68 isolates had similar levels of binding; however, the recent isolates produced 1-2-log higher virus progeny yields than Fermon due to increased replication. We then utilized RNA-seq to define the transcriptional responses in BECs infected with four recent EV-D68 isolates, representing major phylogenetic clades, and Fermon strain. All the tested clinical isolates induced similar responses in BECs; however, numerous upregulated genes in antiviral and pro-inflammatory response pathways were identified when comparing the response to clinical isolates versus Fermon. These results indicate that the recent emergence in severe EV-D68 cases could be explained by increased replication efficiency and enhanced inflammatory response induced by newly emerged clinical isolates.
Project description:Comparative genomic hybridisation of Streptococcus pneumoniae isolates from a single clonal complex, in order to determine genomic diversity. Isolates were selected from a range of tissue types and serotypes in order to cover the full diversity of the clone, and also in order to try and identify tissue-specific genes
Project description:Comparative genomic hybridisation of Streptococcus pneumoniae isolates from a single clonal complex, in order to determine genomic diversity. Isolates were selected from a range of tissue types and serotypes in order to cover the full diversity of the clone, and also in order to try and identify tissue-specific genes Biological replicates: 19 clonal complex 199 S. pneumoniae isolates. One clonal complex 180 isolate used as an outgroup. Independently grown and isolated. One isolate per array
Project description:ChIP-seq of Isw2 (subunit of Isw2 complex) and Nhp10 (subunit of Ino80 complex) was performed on two independent isolates of FLAG-tagged Isw2 or Nhp10 Sample naming convention: <ChIP target>_<growth condition>_<genotype-replicate>_<strain name>
Project description:Ovine mastitis is defined as the inflammation of the sheep udder, most commonly caused in response to intramammary infections. Based on the occurrence of clinical signs, mastitis is characterized as either clinical or subclinical (SCM). The impact of ovine SCM on the overall sustainability of dairy sheep farms has been documented underpinning the significance of efficient diagnosis. Although SCM can be detected in cows, the performance and the validity of the methods used do not transfer in dairy sheep. This fact challenges the development of evidence-based ovine udder health management protocols and renders the detection and control of ovine mastitis rather problematic. Currently, cell culture-based models are being successfully used in biomedical studies and have also been effectively used in the case of bovine mastitis. The objective of the present study was to culture ovine primary mammary cells for the development of 2D and 3D cell culture-based models for the study of ovine SCM. Cells were infected by mastitis-inducing pathogens mimicking the pathogenesis of SCM as derived by natural intramammary infections. The secreted proteins were subjected to mass-spectrometry resulting in the identification of 79 distinct proteins. Among those, several had already been identified in healthy or mastitic milk, while others were detected for the first time in the ovine mammary secretome. The development of cell-based models for the early detection and the overall study of SCM has the potential to be applicable and beneficial for the udder health management in dairy sheep.