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Investigation of compouds affecting the neoplastic cell changes
Andrš, Martin ; Jun, Daniel (advisor) ; Šinkorová, Zuzana (referee) ; Vávrová, Jiřina (referee)
Radiation and genotoxic drugs after more than 70 years since their discovery still belong to the cornerstones of cancer treatment. However, these types of therapy often suffer from severe adverse effects and resistance caused by DNA repair mechanisms. The basic feature of cancer cells is genome instability, leading to various mutations and upregulated or otherwise defective DNA repair, making the cancer cells vulnerable to additional interference with the DNA damage response (DDR) mechanisms. This is the fundamental idea behind the DDR targeted therapy, which has been thoroughly studied for almost two decades. The main goals of this therapy is an improvement of the efficacy of DNA damaging treatments leading to lesser doses and adverse effects but also enabling selective targeting of defective cancer cells. The development of this area of research was very slow at the onset, but last few years it finally brought the first compound into therapy and several others into clinical trials. Among the plethora of signal and effector proteins involved in DDR, three related kinases ATM (ataxia telangiectasia mutated), ATR (ATM and Rad3-related) and DNA-PK (DNA- dependent protein kinase) play the principal roles in initiation and regulation of signaling pathways in response to DNA double and single strand...
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Investigation of compouds affecting the neoplastic cell changes
Andrš, Martin ; Jun, Daniel (advisor) ; Šinkorová, Zuzana (referee) ; Vávrová, Jiřina (referee)
Radiation and genotoxic drugs after more than 70 years since their discovery still belong to the cornerstones of cancer treatment. However, these types of therapy often suffer from severe adverse effects and resistance caused by DNA repair mechanisms. The basic feature of cancer cells is genome instability, leading to various mutations and upregulated or otherwise defective DNA repair, making the cancer cells vulnerable to additional interference with the DNA damage response (DDR) mechanisms. This is the fundamental idea behind the DDR targeted therapy, which has been thoroughly studied for almost two decades. The main goals of this therapy is an improvement of the efficacy of DNA damaging treatments leading to lesser doses and adverse effects but also enabling selective targeting of defective cancer cells. The development of this area of research was very slow at the onset, but last few years it finally brought the first compound into therapy and several others into clinical trials. Among the plethora of signal and effector proteins involved in DDR, three related kinases ATM (ataxia telangiectasia mutated), ATR (ATM and Rad3-related) and DNA-PK (DNA- dependent protein kinase) play the principal roles in initiation and regulation of signaling pathways in response to DNA double and single strand...
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Changes in protein expression in response to genotoxic stress
Ćmielová, Jana ; Řezáčová, Martina (advisor) ; Hofer, Michal (referee) ; Šinkorová, Zuzana (referee)
Human mesenchymal stem cells from the bone marrow, the dental pulp and the periondontal ligament are known for their high proliferation potential and the ability of self-renewal comparable to the other stem cells. They represent a promising field in the therapy due to their ability to differentiate into many cell types. On the other hand, these cells persist in the organism for a long time and a damage in their genetic material would result in mutation of cell genome. The characterization of these cells under genotoxic stress is therefore important. Ionizing radiation is one of the powerful DNA-damaging stressors causing cell cycle arrest associated with reparation or apoptosis. The aim of our work is to study the effect of ionizing radiation on mesenchymal stem cells from the bone marrow, the dental pulp and the periondontal ligament and the response of these cells to ionizing radiation after ATM kinase inhibition. ATM kinase is thought to be a key molecule reflecting DNA damage, especially double strand breaks. We evaluated molecular mechanisms of the apoptosis induction and the reaction to DNA damage caused by ionizing radiation in mesenchymal stem cells isolated from bone marrow, dental pulp and periodontal ligament. The main result of our work is that after the irradiation these cells do not...
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The Study of the Cell and Molecular Mechanisms of B Cell and Intracellular Pathogen Francisella Tularensis Interaction
Bramborová, Lucie ; Wsól, Vladimír (advisor) ; Krejsek, Jan (referee) ; Šinkorová, Zuzana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Lucie Bramborová Supervisor: Prof. Ing. Vladimír Wsól, Ph.D. Supervisor specialist: RNDr. Zuzana Kročová, Ph.D. Title of Doctoral Thesis: Study of cellular and molecular mechanisms of interactions between B-cells and intracellular pathogen Francisella tularensis F. tularensis is Gram-negative, intracellular pathogen responsible for the zoonotic disease tularemia. Immunity to F. tularensis is largely mediated by T lymphocytes but an important role of B lymphocytes during early stage of infection was previously uncovered. The target host cells for F. tularensis replication are macrophages. After the entry into macrophages, F. tularensis resides in phagosomes and then escapes into the cytosol. It replicates within the cytosol and leads macrophages to apoptosis. Recently, important role of B lymphocytes in innate immunity against F. tularensis was demonstrated. Elkins et al. demonstrated strong early protection of mice that was highly dependent on B cells. Direct interaction of B lymphocytes with F. tularensis has not been described. We detected adhesion of F. tularensis to mice and human B lymphocytes, later the entry of bacteria into cells and morphological changes on mitochondria of host...
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Cadaveric bone marrow transplantation: effects of hypoxia and metabolic starvation on mouse hematopoietic stem cells
Linhartová, Jana ; Šefc, Luděk (advisor) ; Průcha, Miroslav (referee) ; Šinkorová, Zuzana (referee)
Objectives: Hematopoietic stem cell transplantation (HSCT) is a widely used method for treatment of hematological disorders and some other diseases. However, sometimes a suitable donor of hematopoietic stem cells (HSC) is not found for a patient. Because HSC have been described as cells with low proliferative and metabolic activity, their tolerance to the lack of oxygen or metabolic substrates may be assumed. In this study, we explored cadaveric bone marrow as an alternative source of HSC for HSCT, using a mouse experimental model. In addition, the effect of in vitro metabolic inhibition and short-term in vitro storage (1 - 4 days) on functional properties of mouse HSC was investigated. Methods: C57Bl/6 mice (wild-type or p53-/- ) were used in the experiments. To explore cadaveric HSC, bone marrow (BM) was left in intact femurs at 37řC, 20řC and 4řC under the conditions of ischemia. The bone marrow cells were harvested after defined time periods ranging 0 - 48 hours. For metabolic inhibition, the electron transport chain inhibitor potassium cyanide (KCN) and inhibitor of glycolysis 2-deoxy-D-glucose (2-DG) were used in vitro. To determine the impact of ischemia, metabolic inhibition, or in vitro storage on transplantability of HSC, the competitive repopulation assay using Ly5.1/Ly5.2 congenic model...
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