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The influence of anesthesia on the degree of DNA oxidative damage
Zubáňová, Veronika ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Background: Oxidative damage is one of the most frequent types of cell components damage leading to oxidation of lipids, proteins and the molecule of DNA. As a consequence, there is a higher occurrence of several pathologies such as atherosclerosis, neurodegenerative diseases, cancer; or diabetes. In our study, influence of whole body anesthesia during minor surgery on the level of DNA damage was examined using comet assay technique. Methods: The basic principle of this method is fixing the cells (lymphocytes) in agarose, their lysis for the removal of membranes, incubation with the specific enzymes and electrophoresis of the released cell nuclei. During the electrophoresis, free low-molecular weight and negatively charged fragments of DNA move towards anode which causes the formation of the typical comet cell shape. Finally, the gels are stained by ethidium bromide (DNA intercalating dye) and visualized. Results: We have observed single strand breakages (SSBs) and, with the use of modified assay using specific enzymes for detection of specific lesions, also oxidized purines and pyrimidines. The extent of DNA damage as determined by the intensity of the tail of the comet was quantified using LUCIA Comet Assay (Laboratory Imaging, Czech Republic) software for image analysis. The results were used...
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Mitochondrial dysfunction and neurodegenerative diseases
Novotná, Veronika ; Herink, Josef (advisor) ; Kovařík, Miroslav (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Veronika Novotná Supervisor: doc. MUDr. Josef Herink, DrSc. Title of diploma thesis: Mitochondrial dysfunction and neurodegenerative diseases The diploma thesis deals with mitochondrial dysfunction and neurodegenerative diseases and it is divided into two main parts. The first part summarized the classification of neurodegenerative diseases and general charakteristic of mitochondria.Then a describe of the processes of oxidative stress, excitotoxicity, apoptosis and briefly decribe the nervous system. The second part deals with description of mitochondrial dysfunction in selected nerodegenerative diseases. The recent studies refer to connection between mitochondrial dysfunctions and formation of neurodegenerative diseases. Keywords: excitotoxicity, mitochondrial dysfunction, neurodegenerative disorders, neuronal cell death, oxidative damage
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The influence of critical condition of patients on DNA damage
Verešpejová, Natália ; Kuchařová, Monika (advisor) ; Bárta, Pavel (referee)
The first cases of patients with pneumonia which grew into an acute respiratory distress syndrome and caused breathing problems began to appear in December 2019. Coronavirus disease 2019 (COVID-19) is the cause of a global pandemic and it is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A complex interplay of factors is responsible for the progression of the disease. Some studies suggest that it promotes oxidative stress and thus may lead to oxidative damage to cells and DNA. The purpose of this study was to observe the relationship between oxidative DNA damage and a critical condition caused by COVID-19 using a comet assay technique. The basic principle of the used method consists in fixation of lymphocytes in an agarose gel, removal of the membrane and cytoplasm of cells, incubation with specific enzymes and electrophoresis. In the process of electrophoresis, negatively charged DNA fragments migrates towards the anode and the cell thus acquires the typical shape of a comet. Comets are visualized using the DNA intercalation dye ethidium bromide. We quantified single - strand breaks and oxidized pyrimidines and purines by using specific enzymes (modification of the method for detecting specific lesions). Results are reported as % tail DNA, thus the percentage of DNA in the...
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The influence of anesthesia on the degree of DNA oxidative damage
Zubáňová, Veronika ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Background: Oxidative damage is one of the most frequent types of cell components damage leading to oxidation of lipids, proteins and the molecule of DNA. As a consequence, there is a higher occurrence of several pathologies such as atherosclerosis, neurodegenerative diseases, cancer; or diabetes. In our study, influence of whole body anesthesia during minor surgery on the level of DNA damage was examined using comet assay technique. Methods: The basic principle of this method is fixing the cells (lymphocytes) in agarose, their lysis for the removal of membranes, incubation with the specific enzymes and electrophoresis of the released cell nuclei. During the electrophoresis, free low-molecular weight and negatively charged fragments of DNA move towards anode which causes the formation of the typical comet cell shape. Finally, the gels are stained by ethidium bromide (DNA intercalating dye) and visualized. Results: We have observed single strand breakages (SSBs) and, with the use of modified assay using specific enzymes for detection of specific lesions, also oxidized purines and pyrimidines. The extent of DNA damage as determined by the intensity of the tail of the comet was quantified using LUCIA Comet Assay (Laboratory Imaging, Czech Republic) software for image analysis. The results were used...
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Mitochondrial dysfunction and neurodegenerative diseases
Novotná, Veronika ; Herink, Josef (advisor) ; Kovařík, Miroslav (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Veronika Novotná Supervisor: doc. MUDr. Josef Herink, DrSc. Title of diploma thesis: Mitochondrial dysfunction and neurodegenerative diseases The diploma thesis deals with mitochondrial dysfunction and neurodegenerative diseases and it is divided into two main parts. The first part summarized the classification of neurodegenerative diseases and general charakteristic of mitochondria.Then a describe of the processes of oxidative stress, excitotoxicity, apoptosis and briefly decribe the nervous system. The second part deals with description of mitochondrial dysfunction in selected nerodegenerative diseases. The recent studies refer to connection between mitochondrial dysfunctions and formation of neurodegenerative diseases. Keywords: excitotoxicity, mitochondrial dysfunction, neurodegenerative disorders, neuronal cell death, oxidative damage
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The influence of anesthesia on the degree of DNA oxidative damage
Zubáňová, Veronika ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Background: Oxidative damage is one of the most frequent types of cell components damage leading to oxidation of lipids, proteins and the molecule of DNA. As a consequence, there is a higher occurrence of several pathologies such as atherosclerosis, neurodegenerative diseases, cancer; or diabetes. In our study, influence of whole body anesthesia during minor surgery on the level of DNA damage was examined using comet assay technique. Methods: The basic principle of this method is fixing the cells (lymphocytes) in agarose, their lysis for the removal of membranes, incubation with the specific enzymes and electrophoresis of the released cell nuclei. During the electrophoresis, free low-molecular weight and negatively charged fragments of DNA move towards anode which causes the formation of the typical comet cell shape. Finally, the gels are stained by ethidium bromide (DNA intercalating dye) and visualized. Results: We have observed single strand breakages (SSBs) and, with the use of modified assay using specific enzymes for detection of specific lesions, also oxidized purines and pyrimidines. The extent of DNA damage as determined by the intensity of the tail of the comet was quantified using LUCIA Comet Assay (Laboratory Imaging, Czech Republic) software for image analysis. The results were used...
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