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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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Bioactive substances of rose hip
Ďubašáková, Mária ; Skoumalová, Petra (referee) ; Veselá, Mária (advisor)
The aim of this master´s thesis was to determine the concentration of bioactive substances such as total polyphenols and flavonoids as well as the total antioxidant capacity in tea extracts from Rosa canina. In the second part, an experiment was found to detect antimicrobial activity and an effective concentration of ethanol against selected bacteria. The theoretical part is focused on description of natural antioxidants and their effect on free radicals, description of tea, description of rose hazel and possibilities of spectrophotometric determination of natural substances and eventually basic distribution of bacteria. In the experimental part, ethanol and aqueous extracts were prepared in which the total number of polyphenols, flavonoids and antioxidant effects. The results of the determination of polyphenols and flavonoids show that the solvent with ethanol is more suitable for flower fermentation and water is more suitable for dropping the rose. Comparison of the antioxidant activity of macerates showed no significant differences; slightly higher antioxidant activity showed macerates from pink flowers. The extracts showed antimicrobial activity against Micrococcus luteus and slightly against Bacillus cereus. The highest antimicrobial activity was determined for the micrococcus luteus
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Lipid composition of biological membranes as a pace-maker of metabolic rate and life-histories
Šimonová, Kateřina ; Tomášek, Oldřich (advisor) ; Vojtek, Libor (referee)
Slože í iologi ký h e rá á zásad í vliv a pol e as e é ast é k seli , jako je apříklad DHA, jsou vel i itlivé k poškoze í vol ý i radikál , oproti o o e as e ý MUFA a as e ý SFA ast ý k seli á . předpokladu, že spolu s e as e ostí e rá se zv šuje o idač í u poškoze í a ásled ě r hlost stár utí, vz ikla a e akerová teorie stár utí Tato h potéza je slože á ze dvou částí. Prv í z týká souvislosti ezi slože í e rá a délkou života, kd živoči hové s ví e e as e ý i e rá a i s v šší pero idač í i de e a počte dvoj ý h vaze ěli ít, dík v šší itlivosti k o idač í u kratší délku života Tato část ašla této rešerši podporu. Druhá část týkají í se slože í e rá a r hlosti eta olis u, la větši ou studií v vrá e a. Me rá ů v šší r hlostí eta olis u spíše é ě e as e é oproti e rá á živoči hů po alejší eta olis e , se e ašla sig ifika t í korela e ož h potézou.
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Physiological adaptations associated with body size and longevity in endothermic vertebrates
Pazdera, Lukáš ; Tomášek, Oldřich (advisor) ; Blažek, Radim (referee)
Abstact Lifespan is related to a number of physiological processes that are interconnected. Longevous species are assumed to have evolved certain physiological adaptations that slow down ageing rate and enable long lifespan. The aim of my thesis was to review physiological traits that has been hypothesised to be associated with lifespan, and to evaluate the empirical support for such hypotheses. Studies testing the association of these traits with body size has also been included in my thesis, as body size is known to be tightly related to lifespan. Traits that are reviewed in my thesis due to their hypothesised coevolution with lifespan are as follows: metabolic rate (i.e., a total amount of energy expended by the organism per unit of body mass and unit of time), oxidative stress resistance, telomere length and rate of telomere shortening and intensity of stress and immune responses. Key words: Lifespan, longevity, survival, reproduction, oxidative stress, free radicals, reactive oxygen and nitrogen species, metabnolic rate, immunity, antioxidants
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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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Impairment of cellular metabolism as common pathophzsiological mechanism of CNS diseases
Hasala, Ondřej ; Otáhal, Jakub (advisor) ; Konopková, Renata (referee)
Name of thesis: Impairment of cellular metabolism as common pathophysiological mechanism of CNS diseases Problem definition: Every human cell needs energy for living. If the prodcution of ATP (as an universal energy carrier) is broken, cell restricts its activity first and during longterm depletion of ATP, dies. It was found, that cellular metabolism is broken in most pathologies in CNS. Disorder of respiratory chain by free radicals is the best known at Parkinson's disease, epilepsy, brain ischemia etc. Mitochondria, where respiratory chain is situated, is not only the aim of free radicals, but it is their major producer. The activity of respiratory chain decreases during the life and this phenomenan is called aging. Aim of thesis: To determine whether there is increased production of free radicals in mitochondria of rat (LE Wistar) hippocampus during the epileptic seizure. Method: Thesis involves experiment which was done with acute rat hippocampal slices. To induce epileptic seizure it was used 4-aminopyridine model. It was used fluorescence imaging as imaging method. Changes of superoxide production was detected with MitoSOX. Electrophysiological record was taken by programme Spike 2 with stimulation and recording electrode inside the slice. Results: There was no significant difference between...
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Oxidative stress as the possible outcome of opioid treatment
Böhmová, Barbora ; Novotný, Jiří (advisor) ; Žurmanová, Jitka (referee)
Free radicals and reactive oxygen species are continuously formed in all aerobic organisms. They may be implicated in ethiology of some diseases such as cancer, atherosclerosis, rheumatoid arthritis, liver, renal, cardiovascular, and neurodegenerative diseases and premature aging. Free radicals and reactive oxygen species can also cause oxidative damage to important cellular components - proteins, lipids, and DNA. Cells are equipped with antioxidant systems to neutralize these radicals. The antioxidant system includes some vitamins (ascorbic acid, α-tocopherol, β-carotene) and enzymes, e.g., superoxide dismutase converting superoxide radical to hydrogen peroxide and molecular oxygen or catalase destroying H2O2. Another important H2O2-removing enzyme in human cells is glutathione peroxidase. Reduced activity of antioxidants leads to excessive production of free radicals resulting in oxidative stress. Opioids belong among pro-oxidants (chemicals inducting oxidative stress). Opioids can decrease activity of antioxidant enzymes and generate reactive oxygen species. The aim of this bachelor thesis is to gather information regarding the opioid ability to elicit oxidative stress. It briefly summarizes our current knowledge about opioids, oxidative stress, free radicals and antioxidants. Key words Opioids,...
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Effect of CO2 on free-radical damage
Dušková, Eliška ; Kalous, Martin (referee) ; Wilhelm, Jiří (advisor)
CO2, known for its role in acid-base regulations, is a mediator of peroxynitrite-induced oxidative damage and it also enhances Cu,Zn-SOD's antioxidant activity. These two means, by which CO₂ affects free-radical damage, are well-explored, but there are many other mechanisms. When CO2 joins in free radical reactions, carbonate radical is often produced. The carbonate radical specifically damages substrates, but it is not possible to generalize, if the effect is pro-oxidant or antioxidant. A protective role of CO2 has been observed during lipid peroxidation and during peroxynitrite-induced oxidation of DNA, when the carbonate radical caused injury to specific bases, but in the same time it prevented DNA strand breaks. Similarly, CO2 prevented peroxynitrite- induced protein fragmentation as well as it caused injury to specific aminoacids. These observations are mostly based on experiments in a chemical system, which means under simplified conditions. In vivo, CO2 exerts much more mechanisms to affect free radical reactions. Under more complex conditions, as cell culture is, there was an increase of oxidative stress after CO2 exposure. Increased concentration of CO2 causes a change in erythrocyte's function and an increase of oxidative stress on the organism's level.
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Assay of fluorescent products in erytrocytes of patients with Alzheimer' s disease
Kohutiar, Matej ; Wilhelm, Jiří (advisor)
Free radicals are highly reactive species with one unpaired electron in orbital. Reactive oxygen and nitrogen species count among important biologic molecules of radical nature. It is very important to fix a concentration of free radicals in cell on non-toxic limits. Products of radical damage are cumulated extra or intracellulary and they are main components of lipofuscin-like pigments. Lipofuscin-like pigments contains in their molecular structure fluorofores, so they are good substrates for fluorescent analysis. Alzheimer's disease is a very actual social and economical problem. Etiology of Alzheimer's disease is still unknown. Histologically, the characteristic presence of Alzheimer's disease is a senil plaques of amyloide ?. ROS and RNS diffuse through hematoencephalic barrier in vessel's lumen and attacks red blood cells. Radical damage of erythrocytes is associated with an increase of concentration of oxidative stress products in cytosol. Sample for fluorescent analysis has been prepared from a red-blood cell extract from 30 patients and 8 healthy controls. Fluorescent spectra of healthy controls have emission maxima in area 327-343nm. In compare with controls, spectra of pacients were more heterogenous in area upon 380nm. The study of oxidative cell damage is important for understanding of...
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