Translocation of outer membrane vesicles from enterohemorrhagic Escherichia coli O157 across the intestinal epithelial barrier

Outer membrane vesicles (OMVs) carrying virulence factors of enterohemorrhagic Escherichia coli (EHEC) are assumed to play a role in the pathogenesis of life-threatening hemolytic uremic syndrome (HUS). However, it is unknown if and how OMVs, which are produced in the intestinal lumen, cross the intestinal epithelial barrier (IEB) to reach the renal glomerular endothelium, the major target in HUS. We investigated the ability of EHEC O157 OMVs to translocate across the IEB using a model of polarized Caco-2 cells grown on Transwell inserts and characterized important aspects of this process. Using unlabeled or fluorescently labeled OMVs, tests of the intestinal barrier integrity, inhibitors of endocytosis, cell viability assay, and microscopic techniques, we demonstrated that EHEC O157 OMVs translocated across the IEB. OMV translocation involved both paracellular and transcellular pathways and was significantly increased under simulated inflammatory conditions. In addition, translocation was not dependent on OMV-associated virulence factors and did not affect viability of intestinal epithelial cells. Importantly, translocation of EHEC O157 OMVs was confirmed in human colonoids thereby supporting physiological relevance of OMVs in the pathogenesis of HUS

EU Structural Funds and priorities of financing South Bohemian Region

The subject of my diploma work is the {\clqq}EU Structural Funds and priorities of financing South Bohemian Region``. The thesis are focused on the South Bohemian Region and its unused potential and primary objectives that need to be targeted and that could be funded from actual programme period 2007 - 2013. Obtaining these financial sources should lead to further developement of the region as a whole and finaly improve the life of the South Bohemian citizens. The theoretical part of the thesis define the basis and main purpose of the economic policy and social cohesion in its entire context. Programme tools that can help to implement projects supporting the development a strengthening the competitiveness of the region are further defined. The aim of this study is to evaluate the possibilities that the South Bohemian Region can use for obtaining financial sources from structural funds alocated by the EU for the Czech Republic on individual priorities, which were evaluated as important and were stated in the basic, medium-term document - Regional Development Programme (PRK). The evaluation shows that the South Bohemian Region in its basic document (PRK) has defined a priority, which corresponds with the intentions of the European Community, which can be documented by the fact that especially in these areas European Union intend to provide its help. This is a good chance for the Region to fulfill the strategic goals and measures of the region. These goals are met largely through thematic operational programs that are available in current programming period and that correspond with the area of the financial support in the Czech Republic. Solving environmet problems can be given as good example how the EU funds are used. It was confirmed that priority issues can be successfully financed from the operational program for selected areas of environment according to priority axes. Programs in the total volume of 50,6 milliards CZK have already been approved for Czech Republic, 4 milliards CZK were allocated to the South Bohemian Region., It is apparent that the proportion is equivalent to the size of the region. Sufficient use of European financial sources significantly helps to fulfil the goals of the Regional Development Programme

Direct PCR Detection of Burkholderia cepacia Complex and Identification of Its Genomovars by Using Sputum as Source of DNA

We developed a nested PCR assay that detects the recA gene of the Burkholderia cepacia complex in sputum. The product of the first PCR round is also used to identify the genomovar of the pathogen. The protocol achieves high sensitivity and specificity with simple interpretation of genomovar status