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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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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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The role adipose tissue in development of insulin resistance and other metabolic disorders in patients with pheochromocytoma
Klímová, Judita ; Petrák, Ondřej (advisor) ; Kříž, Jan (referee) ; Mlíková Seidlerová, Jitka (referee)
Pheochromocytoma and functional paraganglioma (PPGL) are rare neuroendocrine tumors characterized by catecholamines overproduction, which give a rise to disorders of glucose, lipid, and energy metabolism. The role of adipose tissue in these processes remains unclear. Our aim was to determine the gene expression profile in subcutaneous and visceral adipose tissue of patients with PPGL focusing on endocrine functions of adipose tissue, occurrence of brown (BAT) and beige adipose tissue (BeAT), all in connection with other measured metabolic and energy parameters and levels of circulating adipokines. We demonstrate signs of UCP1-dependent norepinephrine induced thermogenesis connected with overexpression of DIO2 in retroperitoneal VAT of PPGL and higher expression of key transcriptional factors of brown/beige adipogenesis, namely PPARGC1α, CEBPB and PRDM16. However, classic murine BAT or BeAT gene signature in VAT of PPGL was not detected. In subcutaneous adipose tissue (SAT) of PPGL we found signs of possible BeAT transformation, however without simultaneously undergoing UCP1-dependent thermogenesis. We also demonstrate that patients with PPGL have higher serum levels of FGF21 compared to healthy controls and these levels do not differ from obese patients. Furthermore, successful tumor removal...
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The role of energy metabolism in cardioprotection induced by the adaptation to chronic hypoxia
Kolář, David
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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Výběr potravin ve výživě sportovců
Mašková, Dominika
This bachelor thesis is focused on a food selection in sport nutrition, especially on differences in nutrients and energy needs in strength and endurance sports. The first two chapters are devoted to principles of rational nutrition and basic components of nutrition. The third chapter describes energy metabolism during exercise. The other chapters include specifics in the intake of basic nutrients in sport nutrition, fluid intake including. Furthermore in these chapters are described the differences in food timing and choices before, during and after exercise according to the differences between endurance and strength sports. The sixth chapter is devoted to an issue of functional and fortified foods. The last chapter's aims are food supplements and their use in sport nutrition.
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Doplňky stravy pro sportovce
Bartl, Martin
This bachelor thesis focuses on the issue of rational nutrition, nutrition of athletes and nutritional supplements for athletes. First chapter is about dealing with the basics and principle of rational nutrition and basic food components, where I describe the composition, effects and distribution. In the second chapter I focus on the energy metabolism, where I look at nutrition from the energy point of view. The third chapter contains the specifics of intake of basic nutrients in sports nutrition. The last chapter deals with selected nutritional supplements for athletes. I focus on the main effects, target groups of users, appropriate dosage and composition of food supplements.
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Genomics of Preaxostyla flagellates
Novák, Lukáš ; Hampl, Vladimír (advisor) ; Hirt, Robert (referee) ; Van der Giezen, Mark (referee)
Protists inhabiting oxygen-depleted environments have evolved various adaptation to thrive in their niches, including modified mitochondria to various degrees adapted to anaerobiosis. The most radically altered forms of these organelles (Mitochondria-Related Organelles, MROs) have completely lost their genomes and other defining features of canonical aerobic mitochondria. Anaerobic protists are often found as endobionts (parasites, mutualists, etc.) of larger organisms. The endobiotic lifestyle combined with anaerobiosis poses another source of evolutionary pressure forcing unique adaptations in the endobionts. Here we present new insights into the adaptations of an anaerobic protistan phylum Preaxostyla, especially with regard to the reductive evolution of mitochondria, which, uniquely among all known eukaryotes, led to a complete loss of the organelle in the oxymonad Monocercomonoides exilis. We have obtained M. exilis genomic assembly of good quality and completeness, as well as genomic and transcriptomic data of varying quality and completeness from 9 other Preaxostyla species. Based on extensive, thorough gene searches and functional gene annotation on these datasets, as well as phylogenetic analyses and protein localization experiments, we conclude: 1) M. exilis has completely lost the...
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The role of the m6A pathway in the regulation of brain ontogenesis in the rat
Tabáková, Petra ; Telenský, Petr (advisor) ; Holzerová, Kristýna (referee)
N6-methyladenosine (m6A) is the most ubiquitous post-transcriptional RNA modification and has an important role in determining the fate of mRNA transcripts. Among the key proteins of the m6A pathway are methyltransferases (METTL family enzymes), demethylases (FTO, ALKBH family enzymes), and m6A binding proteins (e.g., YTHDF family) which recognize RNA sequences depending on the amount and localization of m6A in target transcripts and subsequently influence the fate of mRNA transcripts. The role of methyltransferases and demethylases is to provide a dynamic balance of m6A levels and possibly to convey mechanisms of specificity for these so-called epitranscriptomic marks, which are not yet fully understood. The main objective of this work was to determine the relative changes in the expression of key m6A pathway proteins during early postnatal development and adulthood in the rat brain. We found that the level of expression of key m6A pathway proteins decreases from birth to adulthood, with the exception of a transient increase between postnatal days 10 and 18. During this period, we also found significant changes in the expression of respiratory chain complexes. However, further research is needed to provide evidence of a mechanistic link between the m6A pathway and brain energy homeostasis during...
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Differences in nutrition of Czech national team in sport climbing and competitive climbers
Kuncířová, Veronika ; Vilikus, Zdeněk (advisor) ; Petráková Doležalová, Radka (referee)
This master thesis evaluates the nutrition of Czech representation in sport climbing. The main goal is the monitoring of quantitative and qualitative composition of the Czech national team in sport climbing and comparing the results with the average group of climbers. The thesis is divided into two parts. The theoretical part deals with the morphological and functional characteristics of a sports climber. It also discusses the specific nutritional requirements related to this sport. The practical part consists of evaluation of the data, which were obtained on the basis of anthropometric measurement, short questionnaire, and five-day dietary record. Anthropometric measurements compare the body compositions of respondents. The aim of questionnaires is to evaluate the interest of individuals in nutrition and type of climbing training. Dietary records monitor the energy balance and the nutritional composition of the food. Eating records were evaluated in Nutri Pro Expert software. Results from both groups were compared with each other and also with the recommendations for the population. The purpose was to find out whether the nutrition recommendations of the Czech representation were fulfilled and also to describe the main differences in diet between top athletes and the average climbing group. The...
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The role of energy metabolism in cardioprotection induced by the adaptation to chronic hypoxia
Kolář, David
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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