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Detection of cells in confocal microscopy images
Hubálek, Michal ; Štursa, Dominik (referee) ; Škrabánek, Pavel (advisor)
The goal of the thesis was to create an application that automatically detects healthy cardiomyocytes from images captured by a confocal microscope. The thesis was created based on the specific needs of researchers from the Slovak Academy of Sciences.The application will facilitate and increase the efficiency of their research,because until now they have to evaluate the images and search for suitable cells manually. The RetinaNet convolutional neural network is used for detection and has been implemented in a user-friendly desktop application. The application also automatically records and stores coordinates of detected cells which can be used for capturing cells in higher image quality. Another advantage of the developed application is its versatility, which allows to train detection on other data, making it applicable to other projects. The result of this work is a functional, standalone and intuitive application that is ready to be used by researchers.
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Evaluation of viability of cardiomyocytes
Kremličková, Lenka ; Čmiel, Vratislav (referee) ; Odstrčilík, Jan (advisor)
The aim of this diploma thesis is to get acquainted with the properties of image data and the principle of their capture. Literary research on methods of image segmentation in the area of cardiac tissue imaging and, last but not least, efforts to find methods for classification of dead cardiomyocytes and analysis of their viability. Dead cardiomyocytes were analyzed for their shape and similarity to the template created as a mean of dead cells. Another approach was the application of the method based on local binary characters and the computation of symptoms from a simple and associated histogram.
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Electrostimulation method for cardiomyocyte life extension
Čermáková, Adéla ; Ronzhina, Marina (referee) ; Čmiel, Vratislav (advisor)
This work deals with electrostimulation cardiomyocytes and design stimulation device for their cultivation and stimulation. First part of work deals with electrophysiological myocytes and the next problems with electrostimulation and preserved their original morphology and physiology structure. The following section is designed and constructed stimulation device to improve cell viability. Stimulation took place under different conditions and the success rate depends on the number of cells that survived.
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Hif1a role in cardiovascular function and heart disease
Matějková, Kateřina ; Pavlínková, Gabriela (advisor) ; Holzerová, Kristýna (referee)
Hypoxia inducible factor 1 alpha (HIF1A) is a transcriptional factor, which plays a central role in the maintenance of homeostasis under hypoxic conditions. It regulates a wide variety of genes encoding proteins that influence metabolism, extracellular matrix composition, oxidoreductase activity or angiogenesis in response to reduced oxygen levels. When HIF1A protein function is impaired, the organism is unable respond appropriately to hypoxia. Altered HIF1A regulation can result in severe tissue damage and eventually lead to death. The heart, as an organ with a huge oxygen consumption, is susceptible to various pathologies caused by hypoxic stress. The role of HIF1A in the heart is rather ambiguous and remains to be elucidated. It plays a role in cardioprotective mechanisms but also promotes the development of inflammation and apoptosis. This thesis aims to clarify the role of HIF1A in maintaining physiological functions of the heart during adaptation to hypoxic conditions using a mouse model with heterozygous Hif1a+/- deletion. Experiments involving molecular and cell biology methods performed on left ventricular tissue were preceded by bioinformatic analysis of data obtained by RNA sequencing of isolated cardiomyocytes. RNA sequencing data were analyzed using the R scripting language (packages...
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Circadian regulation of the cardiovascular system
Nováková, Jessica ; Bendová, Zdeňka (advisor) ; Galatík, František (referee)
Circadian system consist of the main circadian oscillator and the peripheral oscillators. Their function is to enable the adaptation of living organisms to the diurnal environmental changes. Circadian system regulates rhythmical expression of genes in nearly every cell of the body, thereby affecting both the physiological and behavioural processes of mammals and other animals. This bachelor's thesis summarizes a knowledge from experiments which led to the demonstration of circadian rhytms in cardiovascular system. To introduce the topic, the first discoveries of circadian rhythms in the cardiovascular system are mentioned. Then, we focus on the circadian rhythms in cardiomyocytes, vascular endothelial cells and in vascular smooth muscle cells. Another focal point of my thesis is the role of transcription factor Krüppel-like factor 15. Circadian regulation of the cardiovascular system is important for the proper function of the heart and blood vessels. Circadian disruption can cause severe pathophysiological and cardiovascular conditions, such as ventricular hypertrophy, hypertension, atherosclerosis, myocardial infarction or coronary artery disease. Key words: circadian, clock genes, cardiovascular system, cardiomyocytes, vascular endothelial cells, vascular smooth muscle cells, Krüppel-like factor...
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Detection of cells in confocal microscopy images
Hubálek, Michal ; Štursa, Dominik (referee) ; Škrabánek, Pavel (advisor)
The goal of the thesis was to create an application that automatically detects healthy cardiomyocytes from images captured by a confocal microscope. The thesis was created based on the specific needs of researchers from the Slovak Academy of Sciences.The application will facilitate and increase the efficiency of their research,because until now they have to evaluate the images and search for suitable cells manually. The RetinaNet convolutional neural network is used for detection and has been implemented in a user-friendly desktop application. The application also automatically records and stores coordinates of detected cells which can be used for capturing cells in higher image quality. Another advantage of the developed application is its versatility, which allows to train detection on other data, making it applicable to other projects. The result of this work is a functional, standalone and intuitive application that is ready to be used by researchers.
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Mesenchymal stem cells and their regenerative and immunomodulatory potential
Brychtová, Michaela ; Lysák, Daniel (advisor) ; Horák, Vratislav (referee) ; Raška, Milan (referee)
Mesenchymal stem cells and their regenerative and immunomodulatory potential Abstract Mesenchymal stem cells (MSCs) possess multidirectional regenerative ability, which, together with their immunomodulatory potential, makes them promising cell type for therapy of wide variety of diseases. Despite ongoing research, which proved MSCs application to be safe, reported effect of MSCs administration on patients is not convincingly beneficial yet. In our work we focused on elucidation of MSCs role in regeneration of vital organs, heart and liver, where a large damage is life threatening for patients and any improvement in therapy would save many lives. Similar situation is in Graft versus host disease (GVHD), where MSCs immunomodulatory properties could be beneficial. Role of MSCs in heart regeneration was examined in vitro. Primary adult swine cardiomyocytes (CMCs) were co-cultured with or without swine MSCs for 3 days and morphological and functional parameters (contractions, current, respiration) of CMCs were measured. MSCs showed supportive effect on CMCs survival, especially at day 3 of the experiment, where in co-culture was significantly higher number of viable CMCs with physiological morphology and maintained function. Effect of MSCs on liver regeneration was observed in swine model of chronic liver...
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Effect of incubation temperature on growth of the working and conducting myocardium in the embryonic chick
Skuhrová, Kristýna ; Sedmera, David (advisor) ; Gregorovičová, Martina (referee)
It was shown almost 50 years ago that hypothermic incubation of chicken embryos results in a reduction in the size of embryos and an increase in the heart weight, presumably by hypertrophy (increase in cell volume). The chicken embryos were incubated in normothermia (37.5 ř C) and hypothermia (33.5 ř C) from the eleventh embryonic day. On the 17th day, the embryos were weighed and then their hearts were weighed. In agreement with the previous results, hypothermic embryos were 29% smaller and their hearts 18% heavier. The heart-to-body weight ratio was 67% higher in the hypothermic group. The measured cell size was very similar in the target areas and it was also between the two groups. The left ventricle width was twofold that the right one and the difference was not significantly higher in the hypothermia model. Purkinje fibers, the terminal part of the conduction system, were smaller than the working cardiomyocytes. Purkinje fibers were slightly enlarged after hypothermic incubation. The proliferation rate was measured by immunohistochemical labeling of anti-phospho histone H3. The experimental group showed much higher proliferation rate; it reached statistical significance in the right ventricle. Thus, hypothermic incubation resulted in increased growth of embryonic heart based on hyperplasia...
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Mesenchymal stem cells and their regenerative and immunomodulatory potential
Brychtová, Michaela ; Lysák, Daniel (advisor) ; Horák, Vratislav (referee) ; Raška, Milan (referee)
Mesenchymal stem cells and their regenerative and immunomodulatory potential Abstract Mesenchymal stem cells (MSCs) possess multidirectional regenerative ability, which, together with their immunomodulatory potential, makes them promising cell type for therapy of wide variety of diseases. Despite ongoing research, which proved MSCs application to be safe, reported effect of MSCs administration on patients is not convincingly beneficial yet. In our work we focused on elucidation of MSCs role in regeneration of vital organs, heart and liver, where a large damage is life threatening for patients and any improvement in therapy would save many lives. Similar situation is in Graft versus host disease (GVHD), where MSCs immunomodulatory properties could be beneficial. Role of MSCs in heart regeneration was examined in vitro. Primary adult swine cardiomyocytes (CMCs) were co-cultured with or without swine MSCs for 3 days and morphological and functional parameters (contractions, current, respiration) of CMCs were measured. MSCs showed supportive effect on CMCs survival, especially at day 3 of the experiment, where in co-culture was significantly higher number of viable CMCs with physiological morphology and maintained function. Effect of MSCs on liver regeneration was observed in swine model of chronic liver...
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