Host Factors Required for Modulation of Phagosome Biogenesis and Proliferation of Francisella tularensis within the Cytosol

Francisella tularensis is a highly infectious facultative intracellular bacterium that can be transmitted between mammals by arthropod vectors. Similar to many other intracellular bacteria that replicate within the cytosol, such as Listeria, Shigella, Burkholderia, and Rickettsia, the virulence of F. tularensis depends on its ability to modulate biogenesis of its phagosome and to escape into the host cell cytosol where it proliferates. Recent studies have identified the F. tularensis genes required for modulation of phagosome biogenesis and escape into the host cell cytosol within human and arthropod-derived cells. However, the arthropod and mammalian host factors required for intracellular proliferation of F. tularensis are not known. We have utilized a forward genetic approach employing genome-wide RNAi screen in Drosophila melanogaster-derived cells. Screening a library of ∼21,300 RNAi, we have identified at least 186 host factors required for intracellular bacterial proliferation. We silenced twelve mammalian homologues by RNAi in HEK293T cells and identified three conserved factors, the PI4 kinase PI4KCA, the ubiquitin hydrolase USP22, and the ubiquitin ligase CDC27, which are also required for replication in human cells. The PI4KCA and USP22 mammalian factors are not required for modulation of phagosome biogenesis or phagosomal escape but are required for proliferation within the cytosol. In contrast, the CDC27 ubiquitin ligase is required for evading lysosomal fusion and for phagosomal escape into the cytosol. Although F. tularensis interacts with the autophagy pathway during late stages of proliferation in mouse macrophages, this does not occur in human cells. Our data suggest that F. tularensis utilizes host ubiquitin turnover in distinct mechanisms during the phagosomal and cytosolic phases and phosphoinositide metabolism is essential for cytosolic proliferation of F. tularensis. Our data will facilitate deciphering molecular ecology, patho-adaptation of F. tularensis to the arthropod vector and its role in bacterial ecology and patho-evolution to infect mammals

Journalistic views on post-violent peacebuilding in Bosnia and Herzegovina

In this chapter we focus on the media portrayal content of a specific ­traumatic event and journalists’ discourse about it in Bosnia and Herzegovina (BIH). Despite the growing role and authority of journalists in shaping our understanding of collective pasts, the possible role of journalists as active agents in contributing to heightening tensions has been marginalized. Analyzing media texts can demonstrate how a “specific, limited truth” about the start of war in BIH is being selected, instrumentalized, and legitimized in the public awareness. Focus on journalists’ perceptions of war and positive post-violence offers an understanding of different views about the start of the war, and guilt. This is why the basic research questions here deal with how journalists in BIH represent the violent past. Specifically, how do they cover a specific traumatic event and what are their perceptions about possibilities of realizing positive post-violence? Research on post-conflict processes looks at the ways in which people attempt to recreate their social fabric in ways appropriated to the changes in their social environment. Thus, the larger question that we are interested in here is whether journalists, like storytellers, frame their stories according to their ethnical belonging and the cultural environment? Furthermore, what media conditions might make possible positive post-violence after violent conflict

No association between coding polymorphism within Exon 4 of the human surfactant protein B gene and pulmonary function in healthy men

The coding polymorphism (rs1130866) within the surfactant protein B gene is known to associate with certain respiratory abnormalities. We investigated, using spirometry and fluorescence-based PCR, whether this variant influenced pulmonary function in healthy, nonsmoking men. We found no association of pulmonary function with genotype at the rs1130866 locus