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Návrh revitalizace vybrané zahrady
ŠAŠKOVÁ, Klára
The aim of this thesis is to create a draft of revitalization of one garden in the village of Nuzbely in the cadastral territory Hroby. The paper is divided into a literary and a practical part. The literary research deals at first with gardens, their planning and revitalization and also with various garden styles in general. Next parts of the research are focused on the function, distribution and planting of the greenery. The practical part includes mapping of the interest location, as well as the current condition of the garden concerned and consequently, a draft solution of the renewal of its functions.
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Preparation of fusion ligands and evaluation of their binding to NK cell receptors
Nepokojová, Tereza ; Vaněk, Ondřej (advisor) ; Grantz Šašková, Klára (referee)
Natural killer cells (NK cells) are an important part of innate immunity. On their surface they express a complex group of receptors that use different signalling motifs to activate or inhibit NK cell cytotoxic activity. NK cells are capable to kill aberrant cells (namely, viral, infected, and tumour cells) by using special cytotoxic mechanisms to trigger apoptosis. The activating receptors recognize tumour or stress-induced ligands, e.g., NKG2D receptor recognizes the MICA ligand and NKp30 recognizes the B7-H6 ligand. Therefore for human immune system it is only natural that cancer cells are destroyed by NK cells. The current therapeutic goals in the treatment of cancer are primarily focused on strengthening the body's own natural ability to fight with cancer and one possible way is stimulation of NK cells to win this deadly fight. In addition to NK cells, antibodies are also widely used for the treatment of cancer, as well as other immune-related disorders. Most of them are monoclonal antibodies, but antibody fragments are getting attention and are being tested more and more in recent years. This work describes the preparation of three bifunctional fusion proteins: B7-H6-L-aHER2, MICA-L-aHER2, and aHER2-L-MICA, which contain immunoligands for the activating receptors of NK cell and VHH fragment...
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Problems of supplementary food sales in schools and school facilities
Šašková, Klára ; Dlouhý, Pavel (advisor) ; Rambousková, Jolana (referee)
This bachelor's thesis deals with the issue of the complementary sale of food products in schools and school facilities, which is, at the moment, regulated by the appropriate statute (the Decree No. 282/2016 Coll., on requirements on food products for which an advertisement is allowed and which can be offered for sale and be sold in schools and school facilities). In the theoretical part of this thesis, the contemporary legislation on the subject-matter is described first. A brief chapter is dedicated to school meals. Subsequent chapters are devoted to requirements set for child food and the question of technological opportunities for food modification. The underlying goal of the theoretical part was to raise awareness of the contemporary alarming state with a growing trend of the obesity occurrence and to provide the reasoning why an emphasis on a proper food, especially during childhood, is so essential and, consequently, why a certain level of legal regulation in this field is desired. In the practical part of this thesis, food products, which satisfy the requirements set forth by the Decree No. 282/2016 Coll., were searched for. The results were projected into tables and they were verbally evaluated in the form of a discussion and they were compared with the requirements, which are now set forth...
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Preparation of nanoparticles for hepatitis B viral therapy
Kružíková, Zuzana ; Grantz Šašková, Klára (advisor) ; Žáčková Suchanová, Jiřina (referee)
Hepatitis B virus (HBV) represents one of the hot topics of current basic and pharmaceutical research. Although an effective vaccine against HBV exists since 1982, the world prevalence of chronic infection is still alarming. The infection can lead to significant liver damage, often resulting in hepatocellular carcinoma. Chronic HBV infection cannot be cured due to the fact that the viral genome persists in the infected hepatocyte hidden from the host immune response as well as from the antiviral treatment. One of the novel approaches aiming for HBV cure suggests that this cccDNA pool could be destroyed using gene editing tools such as CRISPR/Cas9 system. In order to shift this gene editing system to possible medicinal application, CRISPR/Cas9 has to be specifically delivered into the target cell in order to minimize its putative off-target activity. This thesis focuses at first on the design and efficacy testing of new sgRNAs targeting HBV cccDNA and secondly, it describes modular lipid nanoparticles developed specially for delivery of the CRISPR/Cas9 system in the form of RNA. Keywords: hepatitis B virus, CRISPR/Cas9, gene editing, lipid nanoparticles, mRNA delivery, targeted delivery
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Bacteriophage therapy with focus on urinary catheters biofilms
Ludvík, Vojtěch ; Drda Morávková, Alena (advisor) ; Grantz Šašková, Klára (referee)
The term of the bacteriophage therapy denotes the use of viruses for killing bacteria. My thesis refers about the use of the bacteriophage therapy in the process of treating nosocomial infections caused by the urinary tract catheterization. I focus on the bacteria that are found in the catheters' biofilms and on the selection of bacteriophages that will be capable of the enzymatically degradation of the biofilms as well as the lysis of the present bacteria. After the body fluids contact the surface of the catheter, an environment for the evolution of the biofilm begins to evolve, which leads to its fast expansion and to the development of an infection. In the case of improper hygienically measures and unreasonable duration of the catheterization, the incidence of the infections reaches 100%. Because of the presence of the biofilm, the bacteria demonstrate high resistance to antibiotics, which is why the infections often aren't suppressed and may have fatal consequences. If applied, the bacteriophage cocktail and genetically modified bacteriophages can successfully treat the infection and even prevent from its development. Keywords: Bacteriophage therapy, urinary tract infection, catheterization, biofilm, EPS
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Searching for a physiological partner of Ddi2 (DNA damage-inducible protein homolog 2) protein
Kurfürst, Jaroslav ; Grantz Šašková, Klára (advisor) ; Vaněk, Ondřej (referee)
One of the most important cellular processes, essential not only for protein degradation, is the so called ubiquitin-proteasome system. A key player in this system is ubiquitin, a small protein with unique ability to form various chains. Cellular proteins marked for degradation via ubiquitin, are recruited to the proteasome either by a direct interaction with one of the intrinsic proteasomal receptors, or by adaptor proteins. These proteins typically possess ubiquitin-like domain and ubiquitin associated domain that predispose them for delivering ubiquinated proteins to the proteasome. Adaptor protein called Ddi2 differs from other members of this family by possessing additional domain called the retroviral protease like domain. This domain is structurally similar to HIV protease and its proteolytic function has been discovered only recently. Due to the presence of this proteolytic domain one could expect that Ddi2 might be a deubiquitinase. Here we therefore tested the possible cleavage of diubiquitin chains by recombinantly prepared Ddi2 protein. We can conclude that Ddi2 did not show any deubiquitinating activity in given conditions.
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Ddi1-like proteins: a novel family of retroviral-like aspartyl proteases
Šmilauerová, Kristýna ; Grantz Šašková, Klára (advisor) ; Šmahel, Michal (referee)
Ubiquitin-proteasome system is one of the key pathways which maintain cell homeostasis. Its purpose is to degrade damaged, misfolded or unnecessary proteins. It is also involved in multiple other processes such as DNA damage repair, cell cycle control or signaling. The entire system consists of multiple components, which are mutually strictly regulated. Important part of this system is group of so called proteasome adaptor proteins. Their role is to recognize and bind targeted substrates and transport them to the proteasome for degradation. Ddi1-like (abbrev. from DNA damage-inducible protein 1) protein family, a group of proteins with retroviral aspartyl protease-like domain, belongs to proteasome adaptor proteins. Global biological role of this protein family is only partially understood the most studied member is Ddi1 protein from Saccharomyces cerevisiae, and it is thus a subject of active research. This thesis summarizes published information about this protein family, describes its general characteristics and known functions, situates them in the context of cell processes and thereby might suggest the course of further study.
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Structure determination of helical domain of DNA damage-inducible protein 2
Staníček, Jakub ; Grantz Šašková, Klára (advisor) ; Obšil, Tomáš (referee)
Human Ddi2 and his homologues are scarcely explored family of ubiquitin- like/ubiquitin-associated domain proteins (UBL/UBA proteins), containing domain which is structurally related to dimeric aspartyl proteases from retroviruses. The most studied of this family is Ddi1 protein from Saccharomyces cerevisiae, which functions within the ubiquitin- proteasome system. This key cellular system exploits tightly regulated enzymatic apparatus for highly selective posttranslational modifications of proteins with molecules of ubiquitin, which serves as intracellular signal determining the proteins fate, importantly its degradation in many cases. Ddi1 plays a role of proteasome adaptor within this context, helping the recognition of ubiquitylated proteins by the proteasome. Even though the function as a proteasome receptor is possible for human Ddi2 protein as well, its cellular role might have been altered during the evolution. One of the important steps in decoding its purpose in cell is exploration of function of its individual domains. This work focuses on structural study of neighbouring region of this protease domain, where the helix-rich region called HDD domain is located. This region of Ddi2 was cloned, expressed and subjected to the NMR measurements. Structural information based on the NMR data was...
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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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