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Impact of downregulation of gene expression of the peptidase subunit ClpP of the mitochondrial protease ClpXP on structure and function of mitochondria in human cells
Kolařík, Daniel ; Stibůrek, Lukáš (advisor) ; Pecina, Petr (referee)
Mitochondria are some of the most complex organelles of eukaryotic cell. They have their own genome and transcriptional apparatus and maintain several key cellular functions. A substantial part of cellular energetic metabolism happens in the mitochondria, as well as formation of iron-sulfur complexes, synthesis of several key molecules and they are also the essential organelles for the apoptotic pathway. In order to maintain the quality of proteins in their oxidative environment, mitochondria have developed a complex system of proteases that reaches all the mitochondrial compartments that degrade damaged proteins and thus promote mitochondrial turnover. The aim of this work was to characterise function of ClpP subunit of ClpXP matrix protease, which role was not yet extensively investigated in human cells. Therefore, we used RNA-interference to silence expression of ClpP in HEK 293 cells and then we performed rescue experiment during which we reintroduced ClpP in cells. Our results show that the ClpP subunit does not actively participate in apoptotic pathway, nevertheless it is essential for correct assembly of all the respiratory complexes as well as the quality of mitochondria itself. We have also shown that the system of mitochondrial proteases is highly functional and that a lack of ClpP...
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The role of energy metabolism in cardioprotection induced by the adaptation to chronic hypoxia
Kolář, David ; Žurmanová, Jitka (advisor) ; Adamcová, Michaela (referee) ; Bardová, Kristina (referee)
Cardiac energy metabolism is the one of the most complex system in the body. To sustain life, but also to respond quickly to any sudden changes (e.g. running, emotional stress), the heart has developed a unique ability and has become a metabolic "omnivore". At physiological conditions, long chain fatty acids (LCFAs) present the major energetic source for the adult myocardium. However, the cardiac energy metabolism may be compromised during pathophysiological states. One of the most dangerous is, undoubtedly, ischaemia-reperfusion injury with its acute form, myocardial infarction. However, the adaptation to chronic hypoxia has been known for decades for its cardioprotective effect against I/R. Changes of cardiac energy metabolism induced by the adaptation have not been fully explored and the system conceals still too many secrets. This thesis has aimed to determine how adaptation to chronic hypoxia affects the cardiac metabolism of the rat LVs in the following set-ups: 1. The effect of chronic normobaric hypoxia (CNH; 3 weeks, 5500m) during a brief I/R protocol in vitro on the protein kinase B/hexokinase (Akt/HK) pathway, including the expression and phosphorylation of Akt, the expression and localization of HK, the expression of mitochondrial creatine kinase (mtCKS), and the level of Bcl-2 family...
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Change in bodyweight and body composition during the weightlifting training unit and nutrition in strength sports
Vašků, Karolína ; Matoulek, Martin (advisor) ; Petráková Doležalová, Radka (referee)
This bachelor thesis monitors changes in body weight and body composition during training in professional athletes, three women and four men, from the Czech weightlifting team. The theoretical part describes components of the body composition, basic methods and calculations for its assessment. In addition, energy substrates used in weightlifting and sport nutrition basics, including suitable supplements for power athletes, are mentioned. In the practical part, using the bioelectric impedance device, we evaluated the actual body composition and energy needs of individuals in a series of three defined measurements-before training, after training, and after meals. The results of our qualitative research confirm that the energy needs of power athletes are far higher than predicted. We compared the minimum caloric requirement values obtained with the In Body 230 device measurement to those obtained using the Harris-Benedicts predictive equations (HBE), and pointed to significant deviations in basal metabolism calculations and inaccuracy of the results obtained with this equation. In most cases, with HBE, we have experienced undervalue in comparison to the In Body measurement. We have also confirmed the fact that the BMI is not suitable for the weight loss evaluation in weightlifters because of the...
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Circadian system in mitochondria
Šemíková, Johana ; Bendová, Zdeňka (advisor) ; Moravcová, Radka (referee)
The rhythms of circadian clocks throughout our bodies are not governed only by the light/dark cycle, but in many peripheral tissues circadian clocks are reset based on availability of nutrients and timing of food intake. The circadian system responds to changes in the levels of two metabolites (AMP and NAD+ ) that are central to biochemical reactions involved in energy production, storage, and utilization through the metabolic sensors AMPK and SIRT1. The aim of this review is to summarize mechanisms for energetic metabolism known to date that are connected with the regulation of circadian rhythms and explain their function in maintaining their stability. Last but not least, to show possible dysregulation of these mechanisms and their impact on the circadian system.
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Energy metabolism and apoptotsis markers in cold heart aclimated rat.
Pospíšilová, Barbora ; Horníková, Daniela (advisor) ; Knytl, Martin (referee)
Cold adaptation and her effects has been known for many decates. Positive or negative impact depends especially on its length and strength. The lower temperature can very often cause the stress for organism. On the other hand in expreriment with long-term adapatation were found positive consequences on cardiovascular system. We found the lack of studies devoted to the energy metabolism and apoptosis in heart tissue during long-term cold adaptation. In this work we used a model with milder conditions of the adaptation (10žC±1), so there wouldn't be damage of the experimental animals. We compared the resuls betwen control, cold and regressive group of rats. In this expreriment we used methods of electrophoresis and Western blot. The target of the work was found if we can find any differences betwen chosen HIF targeted genes. The next goal was to detect the differences betwen chosen pro-apoptotic and anti-apoptotic markers. Keywords: cold adaptation, heart, energetic metabolism, HIF, apoptosis
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Modulation of RNA demethylase FTO function in SH-SY5Y cells: the effect on insulin signaling and mitochondrial respiration
Čočková, Zuzana ; Novotný, Jiří (advisor) ; Hlaváčková, Markéta (referee)
Aim of this thesis was to observe changes in oxidative metabolism and expression of important neuroenergetic proteins in human neuroblastoma cell line SH-SY5Y due to inhibition of FTO. FTO is a RNA demethylase that uses N6-methyladenosine as substrate. Differences in enzyme expression are connected to broad area of effects involving energy homeostasis. Mitochondria are cellular powerhouses, a key elements in production of energy and metabolic substrates, yet a source of potentially dangerous reactive oxygen species (ROS) and analogous reactive molecules. In order to better understand FTO purpose in neuronal energetic metabolism, we examined mitochondrial respiratory chain. Using high-resolution respirometry we were capable of observing impairment in mitochondrial respiration after FTO inhibition. There was considerable decline in endogenous respiration, maximal respiration rate and reserve capacity. In order to obtain more detailed view into mitochondrial respiration, expression levels of electron-transport complexes were quantified by Western blot technique. Slight reduction was identified in subunits of complex I and IV. However, the most prominent alteration was seen in complex II subunit. There were no differences in expression of complex III and ATP synthase subunits. Beside disrupted activity...
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Monitoring of carbohydrate income in sportsmen's diet
Pourová, Veronika ; Vilikus, Zdeněk (advisor) ; Petráková Doležalová, Radka (referee)
The topic of my master thesis is monitoring of carbohydrate intake in sportsmen's diet. In the thesis, intake of carbohydrates as well as other macronutrients is monitored in order to evaluate, if athletes prefer other energy sources to carbohydrates and if their total energy intake is sufficient. If athletes prefer easily digestible carbohydrates or if they follow recommendation for healthy diet and increase their intake of whole foods, legumes, fruits and vegetables can be evaluated by monitoring fiber and sugar intake. All the monitored data are compared with non-athletes in order to see, in which field their diets vary. In the theoretical part, principles of sports nutrition are defined, basics of population recommendations are mentioned and methods of nutritional assessment and energy expenditure estimation are explained. In the practical part, five-day food-diaries of athletes and non-athletes are evaluated by NutriPro Expert software. Five-day physical activity diaries recorded during the same days are evaluated by using the Compendium of Physical Activities. All the studied values (energy intake, energy expenditure, carbohydrate, fat, protein, sugar, fiber and alcohol intake) were compared to recommended values for athletes and non- athletes and further compared among these two groups. The...
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