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Effect of obesity and metabolic complications on molecular characteristics of bone marrow skeletal stem cells in relation to bone and adipose tissue metabolism
Beňová, Andrea ; Tencerová, Michaela (advisor) ; Kuda, Ondřej (referee) ; Rauner, Martina (referee)
Obesity and diabetes are metabolic complications associated with higher accumulation of fat in periphery and bones leading to higher fracture risk. Several approaches have been introduced in the treatment of obesity and diabetes - non-pharmacological (e.g. dietary intervention or physical activity) and pharmacological (e.g. metformin, thiazolidinediones (TZDs)) to improve metabolic complications. Dietary intervention including omega-3 polyunsaturated fatty acid (PUFA) supplementation showed multiple beneficial effects on bone parameters and bone marrow skeletal stem cells (BMSC) properties in animal and clinical studies. However, this was not well studied in obesogenic conditions. Further, TZDs (e.g. pioglitazone, rosiglitazone) are very effective insulin sensitizers but with several side effects (increased bone marrow adiposity (BMA) with higher fracture risk) due to their mechanism of action. MSDC-0602K, a novel TZD analog, has been developed to minimize side effects on fat metabolism. However, its impact on bone phenotype and bone marrow skeletal stem cells BMSCs in relation to obesity has not been intensively studied. Thus, in my PhD thesis I investigated both approaches using high-fat diet (HFD)-induced obesity mouse model. 8 weeks old male mice were fed with HFD or HFD supplemented with...
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Microplastics and their effects on the metabolism of animal cells
Fiedlerová, Gabriela ; Cajthaml, Tomáš (advisor) ; Černý, Jan (referee)
Microplastics are tiny particles smaller than 5 mm in size found in various environments, including seas, rivers and soil. These particles can be intentionally produced as part of cosmetic products or formed by the breakdown of larger plastic objects. Microplastics represent a serious threat to the environment and animals and could eventually reach humans through the food chain. This thesis deals with a critical and synthetic overview of the literature related to the influence of microplastics on the metabolism of animal cells. The thesis also considers the experimental conditions under which the data were obtained. In particular, factors such as the tested microplastics' shape, size and ageing are critically evaluated as these aspects are often neglected in the literature. Current knowledge shows that microplastics accumulate in the bodies of animals and cause physical and metabolic damage, inflammation, disruption of energy metabolism, protein metabolism and amino acid and lipid metabolism. The most serious effects of microplastics can be considered to be the formation of oxygen radicals, neurotoxicity, disruption of gametogenesis and offspring development.
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Characterization of a role of selected antiapoptotic Bcl-2 family proteins in mitochondrial metabolism.
Antoš, Šimon ; Anděra, Ladislav (advisor) ; Brábek, Jan (referee)
Proteins from the Bcl-2 family are now for over 30 years widely studied mainly for their key role in apoptosis, a principal mode of regulated cell death. In the last ten years Bcl-2 proteins were also linked to the regulation of cellular signaling, mainly cellular metabolism and respiration. In this study we aimed to analyze non-apoptotic function of Bcl-2 proteins by their genetic elimination using the CRISPR-Cas12a approach and by the subsequent analysis of mitochondrial respiration, glycolysis and metabolic profiling. Our results confirmed that Bcl-2 proteins can modulate the level of mitochondrial respiration. The elimination of anti-apoptotic proteins Bcl-2, Bcl-XL and Mcl-1 decreased high respiration of cells lacking pro-apoptotic proteins Bax and Bak to the levels observed in parental U87-MG glioblastoma cells. Therefore, the loss of anti-apoptotic Bcl-2 proteins has greatly impacted mitochondrial respiration and it points to their role in a regulation of oxidative phosphorylation.
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Role of sulfhydryl oxidase 1 in cancerogenesis
Beranová, Lea Marie ; Truksa, Jaroslav (advisor) ; Šuťák, Róbert (referee)
Disulfide bridges play a significant role in protein-folding as well as en- zyme activity and thus regulate many intra- and extracellular processes. Sulfhydryl oxidase QSOX1 forms S-S bridges de novo, modulating the activity of its substrates and thus directly or indirectly influences vital cel- lular processes. The first part of this thesis focuses on characterization of the role of QSOX1 in cancerogenesis, using breast cancer cell lines (MCF7, MDA-MB-231) and pancreatic cancer cell line (Panc-1), while the second part emphasizes the regulation of QSOX1 expression by different oxygen concentrations. To study the effect of QSOX1 on proliferation of triple-negative cancer cells MDA-MB-231, two genetically modified cell lines - QSOX1-overexpressing and QSOX1 knockout cell lines - were constructed. While increased QSOX1 protein levels do not have a significant effect, the absence of QSOX1 leads to a decreased cellular growth. Lack of QSOX1 also results in visible change in cellular morphology. QSOX1 knockout cells can be mostly characterized as more round-shaped with less noticeable or completely missing lamellipo- dia. This finding is with agreement with to-date literature suggesting that QSOX1 is important not only for cellular proliferation but also for migration and invasiveness. While authenticating the theory of...
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Role of sulfhydryl oxidase 1 in cancerogenesis
Beranová, Lea Marie ; Truksa, Jaroslav (advisor) ; Šuťák, Róbert (referee)
Disulfide bridges play a significant role in protein-folding as well as en- zyme activity and thus regulate many intra- and extracellular processes. Sulfhydryl oxidase QSOX1 forms S-S bridges de novo, modulating the activity of its substrates and thus directly or indirectly influences vital cel- lular processes. The first part of this thesis focuses on characterization of the role of QSOX1 in cancerogenesis, using breast cancer cell lines (MCF7, MDA-MB-231) and pancreatic cancer cell line (Panc-1), while the second part emphasizes the regulation of QSOX1 expression by different oxygen concentrations. To study the effect of QSOX1 on proliferation of triple-negative cancer cells MDA-MB-231, two genetically modified cell lines - QSOX1-overexpressing and QSOX1 knockout cell lines - were constructed. While increased QSOX1 protein levels do not have a significant effect, the absence of QSOX1 leads to a decreased cellular growth. Lack of QSOX1 also results in visible change in cellular morphology. QSOX1 knockout cells can be mostly characterized as more round-shaped with less noticeable or completely missing lamellipo- dia. This finding is with agreement with to-date literature suggesting that QSOX1 is important not only for cellular proliferation but also for migration and invasiveness. While authenticating the theory of...
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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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