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The influence of spinal anesthesia on the degree of DNA damage
Koščáková, Mária ; Kuchařová, Monika (advisor) ; Vokřál, Ivan (referee)
Background: The human organism is exposed daily to many endogenous and exogenous substances that are the source of oxidative damage. Cell structures, including DNA (deoxyribonucleic acid) in the nucleus are damaged due to high concentrations of these substances and accumulation of oxidative stress in cells. The predominance of these damaging processes may later be responsible for human diseases such as cancer, neurodegenerative diseases or heart failure. In our study, we observed oxidative damage at the DNA level due to spinal anesthesia. Methods: Sample processing was performed by comet analysis. The principle consists in fixation of cells (lymphocytes) in the agarose gel, lysis of cell structures for nucleotide release, incubation with specific enzymes and exposure to electrophoresis. Damaged, negatively charged parts of the DNA in the electric field are directed to the positive charged anode, creating a typical comet shape. For visualization, the gels were stained with ethidium bromide (DNA intercalating dye). Results: We have quantified single-strand breaks, oxidized purines and pyrimidines (use of enzymes to detect specific damages). The results are reported in percentage of DNA in the comet's tail. The principle is to compare the intensity of the comet's tail with the total comet intensity....
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The influence of spinal anesthesia on the degree of DNA damage
Koščáková, Mária ; Kuchařová, Monika (advisor) ; Vokřál, Ivan (referee)
Background: The human organism is exposed daily to many endogenous and exogenous substances that are the source of oxidative damage. Cell structures, including DNA (deoxyribonucleic acid) in the nucleus are damaged due to high concentrations of these substances and accumulation of oxidative stress in cells. The predominance of these damaging processes may later be responsible for human diseases such as cancer, neurodegenerative diseases or heart failure. In our study, we observed oxidative damage at the DNA level due to spinal anesthesia. Methods: Sample processing was performed by comet analysis. The principle consists in fixation of cells (lymphocytes) in the agarose gel, lysis of cell structures for nucleotide release, incubation with specific enzymes and exposure to electrophoresis. Damaged, negatively charged parts of the DNA in the electric field are directed to the positive charged anode, creating a typical comet shape. For visualization, the gels were stained with ethidium bromide (DNA intercalating dye). Results: We have quantified single-strand breaks, oxidized purines and pyrimidines (use of enzymes to detect specific damages). The results are reported in percentage of DNA in the comet's tail. The principle is to compare the intensity of the comet's tail with the total comet intensity....
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The influence of spinal anesthesia on the degree of DNA damage
Koščáková, Mária ; Kuchařová, Monika (advisor) ; Vokřál, Ivan (referee)
Background: The human organism is exposed daily to many endogenous and exogenous substances that are the source of oxidative damage. Cell structures, including DNA (deoxyribonucleic acid) in the nucleus are damaged due to high concentrations of these substances and accumulation of oxidative stress in cells. The predominance of these damaging processes may later be responsible for human diseases such as cancer, neurodegenerative diseases or heart failure. In our study, we observed oxidative damage at the DNA level due to spinal anesthesia. Methods: Sample processing was performed by comet analysis. The principle consists in fixation of cells (lymphocytes) in the agarose gel, lysis of cell structures for nucleotide release, incubation with specific enzymes and exposure to electrophoresis. Damaged, negatively charged parts of the DNA in the electric field are directed to the positive charged anode, creating a typical comet shape. For visualization, the gels were stained with ethidium bromide (DNA intercalating dye). Results: We have quantified single-strand breaks, oxidized purines and pyrimidines (use of enzymes to detect specific damages). The results are reported in percentage of DNA in the comet's tail. The principle is to compare the intensity of the comet's tail with the total comet intensity....
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L01 DNA damage formation and DNA repair following an intervention of colorectal cell lines with ganoderma lucidum
Vodička, Pavel ; Opattová, Alena ; Čumová, Andrea ; Slíva, D.
Colorectal cancer (CRC) is the third most common malignancy in the world and second most common cause of cancer related deaths in Europe. CRC is complex disease that develops as consequence of environmental and health risk factors with involvement of suboptimal DNA repair, resulting in an accumulation of DNA damage. Reactive oxygen species (ROS) are highly reactive molecules strictly controlled by cellular antioxidant system. Disturbance in the prooxidation–antioxidation homeostasis increases an extent of ROS and consequently an accumulation of DNA damage as well as apoptosis. \nMany natural compounds possess anti-cancer activities tentatively mediated by the generation of ROS. Cancer cells are more sensitive to oxidative DNA damage than non-malignant ones. Modulation of oxidative DNA damage and its repair by natural compounds may lead to selective cancer cell-death and further sensitization of cancer cells to the treatment. Ganoderma Lucidum (GLC) (Reishi, Ling-Zhi), a mushroom used in Chinese medicine for thousands of years, represents an example of a natural compound with empirically recorded anti-cancer as well as anti-proliferative effects. \nThe aim of our study is to define effect of Ganoderma lucidum (GLC) extract on DNA damage and DNA repair system in colorectal cell lines with different genetic backgrounds.\nOur results suggest that GLC extract decreases activity of the cellular antioxidant system which leads to oxidative DNA damage. GLC extract increases genotoxic burden in colorectal cancer cell lines, highlighted by the suppressed base excision repair capacity. These data indicate that specific oxidative DNA damage caused by natural compounds may become a potential tool for the improvement of specific anti-cancer treatment.\n
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