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Effect of mitochondrially targeted senolytic agents on the development of respiratory diseases
Rennerová, Michaela ; Štemberková-Hubáčková, Soňa (advisor) ; Neckář, Jan (referee)
Aging is a complex biological process characterized by gradual decline in the physiological functions of cells and tissues, leading to the activation of a process known as cellular senescence. This results in the development of structural and functional changes in the organism accompanied by an increased risk of various diseases. Respiratory diseases are among the most common chronic conditions associated with the accumulation of senescent cells in the body. Current treatment for chronic respiratory diseases primarily focuses on symptom relief rather than addressing the primary cause of these conditions. Given that senescent cells play a significant role in the pathogenesis of respiratory diseases, the future of therapies lies in their elimination using senolytic agents. Despite promising results from some studies, current research is limited by the heterogeneity of senescent cells, which is reflected in their sensitivity to senolytic agents and the variability of therapeutic responses. Mitochondria play a central role in the development and maintenance of the senescent cell phenotype by regulating key processes such as energy metabolism and cellular signaling, regardless of the type of senescent cell. Thus, targeting mitochondria in senescent cells may represent a promising therapeutic strategy...
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Ketogenic diet as a treatment for inherited disorders of ATP synthase
Kudrnovská, Barbora ; Mráček, Tomáš (advisor) ; Tesařová, Markéta (referee)
The Tmem70 protein plays a significant role in the biogenesis of ATP synthase, mutations in the Tmem70 gene are a common cause of mitochondrial disease and have been associated with neonatal encephalocardiomyopathy. Similar to other mitochondrial disorders, there is currently no available treatment for Tmem70 defects. However, the ketogenic diet (KD) appears to be a potential therapeutic intervention for various mitochondrial disorders, and our preliminary results demonstrate its positive impact in a mouse model of Tmem70 dysfunction. Therefore, using this Tmem70 model, we decided to test a modified KD, which could be a better alternative for a potential application in human patients. The effect of the modified KD was examined in a mouse model of tamoxifen-induced whole-body knockout of Tmem70 (Tmem70 KO). The modified KD led to improved survival, slowed weight loss, and normalization of plasma indicators of liver damage. We detected a positive effect of the modified KD on the assembly of ATP synthase in liver tissue, however, skeletal muscle showed only a minimal response. Based on the results of this study, we infer that although the absence of functional Tmem70 affects both liver and muscle tissues, the greater impact on the phenotypic manifestation in the Tmem70 KO mouse model is due to liver...
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Changes in mitochondrial DNA in colorectal cancer patients
Danešová, Natálie ; Vodenková, Soňa (advisor) ; Büchler, Tomáš (referee)
Mitochondrial DNA (mtDNA) changes occur more frequently in the cell than alterations in nuclear DNA (nDNA) due to factors such as proximity to reactive oxygen species (ROS) or fewer DNA repair pathways. While nDNA damage is known to play a role in colorectal cancer (CRC), there is a lack of studies concerning mtDNA damage. Changes in mtDNA copy number, a possible indirect marker of mtDNA damage known as mtDNA content, have been reported in CRC with conflicting results. Various changes in mtDNA have been observed in multiple cancer types and proposed as potential biomarkers, including CRC; however, its exact role in disease progression, patients' prognosis, or prediction of treatment is yet to be determined. The primary hypothesis of this study was that mitochondrial dysfunctions resulting from mtDNA changes play a role in colorectal carcinogenesis and could serve as potential CRC biomarkers. To test this hypothesis, we measured mtDNA damage, mtDNA content, and the expression of selected DNA repair genes in both tumor and adjacent non-malignant mucosa. Initially, we conducted a Pilot study involving 7 patients, utilizing RNA sequencing and qPCR. Subsequently, a Validation study was performed on a larger cohort of 50 patients using qPCR. Our findings revealed that adjacent mucosa exhibited higher...
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Modulation of mitochondrial transfer by influencing mesenchymal stem cells
Fráňová, Markéta ; Krulová, Magdaléna (advisor) ; Rohlenová, Kateřina (referee)
Mesenchymal stem cells (MSCs) have the ability to modulate the immune response. They use several mechanisms to affect the function of immune cells, and mitochondrial transfer is one of them. Recieving mitochondria from MSCs induces metabolic changes in immune cells, thereby promoting their shift to an anti-inflammatory phenotype. Due to their properties, MSCs have a potencial to be used in therapies, for example in a treatment of autoimmune diseases. The problem of MSCs-based therapies is their low efficacy, mainly due to the high mortality of stem cells after transplantation. In order to achieve at least some effect, the large number of cells is needed for application. The required number of cells can be obtained only by in vitro expansion. However, a long-term culture has a negative impact on MSCs and their immunomodulatory properties. Enhancing MSCs function could increase the efficacy of MSCs-based therapies. The aim of this thesis was to determine whether mitochondrial transfer can be modulated by stimulation of MSCs with selected factors. MSCs were treated with rapamycin, insulin-like growth factor 1 (IGF-1), interferon gamma, or oligomycin. Then the effect of these factors on mitochondria and their transfer to immune cells, metabolism, and immunomodulatory properties of MSCs was analyzed. We...
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The role of exosomes in communication between adipose and cardiac tissue
Novotná, Denisa Regine ; Zouhar, Petr (advisor) ; Kašík, Petr (referee)
Extracellular vesicles, including exosomes, represent an important, newly identified, factor in the information transmission between individual cells and organs of a multicellular organism. Usually, exosomes contain nucleic acids, proteins and even whole organelles such as mitochondria. The exosomes production takes place, among others, in adipocytes of adipose tissue. Because adipose tissue is the main repository of stored lipids, its metabolism and the functionality of its mitochondria react sensitively e.g. to nutritional conditions. Recent studies have demonstrated that exosome secretion from adipose tissue may play a pivotal role in the progression of metabolic syndrome. In hypertrophied tissue, mitochondria fail, and adipocytes dispose of them by secreting them as exosomes. The majority of these exosomes with damaged mitochondria are directly taken up by macrophages within the adipose tissue. However, a subset of them can escape into the circulation and reach the heart. The uptake of these exosomes by the myocardium and the incorporation of damaged mitochondria into its own mitochondrial network may have surprisingly positive effects for increasing resistance to damage in infarction. Furthermore, exosomes may also play an important role in ridding brown adipose tissue and other cells of...
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Analysis of mitochondrial energetic metabolism in early stages of Huntington's disease
Ďurčeková, Katarína ; Hansíková, Hana (advisor) ; Kohoutová, Michaela (referee)
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, psychiatric and cognitive symptoms. The average age of onset of HD is 40 years and affected individuals have an unfavourable prognosis. Since there is still no curative treatment available, the current treatment of HD focuses only on symptom suppression. The clinical manifestation of HD is preceded by an asymptomatic period in which continuous biochemical, structural and functional changes at the cellular level are already occurring. Our previous studies have demonstrated impaired mitochondrial parameters in lymphocytes and cultured fibroblasts of patients with advanced HD. The aim of this study was to characterize the essential components of mitochondrial energy metabolism by functional methods and to find suitable mitochondrial biomarkers to monitor progression of HD. The analysed set consisted of 12 cell lines of cultured skin fibroblasts from patients with genetically proven HD, four patients in asymptomatic, four in early and four in symptomatic stages of HD. Four lines from healthy individuals served as controls. Mitochondrial respiration and glycolytic activity were analysed by real-time monitoring of oxygen consumption and pH changes, ATP production by luminescence, and metabolic turnover...
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Similarity/dissimilarity analysis of genomic data on the basis of graphical representation
Těthal, Jiří ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
The work deals with the identification of species of animal through the density of nucleotids of mitochondrial gene CO1. In the first part the theory summarized information about DNA barcoding and the mitochondria, cellular organel that with this method is closely related. The second part deals with virtually comparing different sequences using the density of nucleotides. For this it was created program, which uses two functions, the first of the specified nucleotide sequence calculates the density and one can compare the density by distance methods.
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Comparison of mitochondrial DNA for species identification
Labounek, René ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
The work deals with the method of recognizing species on the analysis of mitochondrial DNA segment. This analysis and classification using segment gene called CO1 in literatures such as barcode of life. In the beginning of work is analyzed the mitochondrial theory of heredity and conditions of formation of barcode. Practical use is based on this theory in creating database of barcodes generated to different animal species. Data used for creating the library are drawn from public databases NCBI and BOLD Systems. The next part of this work concerns about methods of comparison of the individual barcodes to the others and especially to the barcode of human. Three main computing methods were used tore these analyses: Needleman-Wunsch algorithm, Smith- Waterman algorithm and comparison of similarities using distance matrix. This work also concerns about transformation of DNA molecule sequences from symbols to numeric formats, which is required for the distance matrix comparison method. Algorithms for searching for a barcode of a species and vice versa were created to ease the work with data.
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Computational modelling of mechanical tests of animal cell
Orlová, Lucie ; Fuis, Vladimír (referee) ; Burša, Jiří (advisor)
Předkládaná diplomová práce se zabývá stavbou živých živočišných buněk a jejich odezvou na mechanické zatěžování. Zobecněným zaměřením práce je popis mechanického chování buňky nejenom ve fyziologickém, ale i v patologickém stavu. Výchozím předpokladem pro úspěšné řešení zadané úlohy je vysoce interdisciplinární přístup kombinující výpočtové přístupy mechaniky těles (v~tomto případě metodu konečných prvků) s lékařským výzkumem. Nejdůležitějším bodem při tvorbě výpočtového modelu, pomocí něhož je možné aproximovat chování živé buňky při zatížení, je zejména identifikace mechanicky významných komponent a~jejich materiálových parametrů. V tomto případě jsou jako mechanicky význačné identifikovány spojité součásti jádro, membrána a cytoplazma, které jsou nově propojeny s prvky diskrétními (mitochondriální sítí) v hybridním modelu, jehož platnost je ověřena pomocí experimentálních dat. Tento model slouží jako podklad k vyhodnocení míry vlivu mitochondrií na celkovou tuhost buňky.
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Ferrous nanoparts localization in inner organelles
Solař, Jan ; Janoušek, Oto (referee) ; Čmiel, Vratislav (advisor)
This bachelor‘s thesis describes a behavior of iron nanoparticles in human mesenchymal stem cells. First section deals with methods of fluorescent labelling of organelles and nanoparticles and settings of confocal microscope for their detection. Next section describes the iron nanoparticles metabolism and accumulation in cells. Finally, there is a section about the developement of the software utility for the localization and for the quantitative analysis of the fluorescence organelles and nanoparticles inside the living cells.
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