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Nanoparticles for gene editing
Kružíková, Zuzana ; Grantz Šašková, Klára (advisor) ; Beranová, Jana (referee)
Early DNA-based therapies were tested for therapeutic applications, but they sooner or later revealed multiple hurdles and risks preventing their use in further clinical trials. Recently, they have been replaced by rapidly evolving gene editing using programmed nucleases capable of precise genome modifications by cleaving specific DNA sequences. Zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and CRISPR/Cas9 system are currently under investigation as potential therapeutics. However, their off-target effects must be controlled. Targeted delivery of nucleases in a form of mRNA seems as the most promising method. Various types of nanoparticles enable mRNA transfer and could be used to facilitate the nuclease application. Some of these nanoparticles together with characterization of the programmed nucleases are described in this thesis.
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Interaction of Borrelia sp. with HL-60 cells and monocytes and cultivation of Anaplasma phagocytophilum in HL-60 cell culture
Marková, Lucie ; Hulínská, Dagmar (advisor) ; Beranová, Jana (referee)
Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...
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Microdomains in freshwater sediment as a driving factor in the biogeochemical processes
Duchoslav, Vojtěch ; Falteisek, Lukáš (advisor) ; Beranová, Jana (referee)
A natural arsenic anomaly at Mokrsko is a site of biogenic precipitation of realgar (As4S4) in stream sediment. The extent of the phenomenon in Mokrsko is globally unique. A previous study showed the ability of microorganisms to solubilize arsenic from its secondary minerals and to produce realgar precursors, i.e. sulfides and arsenites, by anaerobic respiration. The study also raised questions about the mechanism of realgar precipitation since physicochemical conditions favoring this reaction were never detected despite significant sampling efforts. We chemically and microbiologically analyzed sedimentary profiles to the depth of ca. 120 cm in order to understand the functioning of the biogeochemical system. The profiles comprised both the unsaturated and the saturated zone. We distinguished six different domains representing environments from the surface soil to anoxic sedimnent containing realgar-encrusted wood. An analysis of phylogenetic dissimilarity revealed that microbial communities from the various domains form distinct clusters. This suggests that different conditions prevail in the various domains, and that different biogeochemical processes take place there. Incubation (microcosm) experiments showed that bioprecipitation of realgar can be conducted in vitro. It requires a suppression...
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Application on nanoparticles in bio-medicine.
Bělinová, Tereza ; Hubálek Kalbáčová, Marie (advisor) ; Beranová, Jana (referee)
During last few years nanoparticles of different origin have been used in biomedicine. Their interactions with cells are however a big point of concern and so further research is needed to be done regarding their properties, internalization into cells and their fate within the cell. All of this is needed for correct selection of proper nanoparticles for bioimaging and also for future use in human medicine. One of the main parts of research is deep understanding of interactions of nanodiamonds and silicon based nanoparticles with cells, because of their highly unique properties (autofluorescence), biocompatibility and in case of silicon nanoparticles also their degradability.
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Internet, public libraries and information policy in Cuba
Beranová, Jaroslava ; Šisler, Vít (advisor) ; Očko, Petr (referee)
The aim of this thesis is to present the development and current state of information policy and public libraries in Cuba. The first part of thesis outlines historical and political development in Cuba since the beginning of the revolution in 1959 to the present and introduces the development of selected areas of information policy on the island. The second part analyzes the expansion of information and communication technologies, including Internet and its censorship. The last part of thesis summarizes historical development and current state of public libraries and partially librarianship.
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