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Group of specialized cells that form cardiac conduction system is responsible for generation and coordinated propagation of the electrical impulse in the heart. Changes in its development can be connected with arrhythmias; therefore, a good level of knowledge is necessary and relevant for basic science and clinical practice. For correct development of the conduction system are important genes coding gap junctions proteins, ion channels, transcription factors and other molecules involved in signaling cascades (endothelin, neuregulin). Development of conduction system is determined in addition to genetic factors also by epigenetics and environmental factors. This thesis with its individual papers on which it is based is addressing different aspects of conduction system development, which appears to be a complex process. Another feature which is linking all papers together, is the methodological approach enabling us to study function of the conduction system - optical mapping. In the first publication we studied by the means of in vitro organ culture the impact of work load without interfering hemodynamics on the conduction system maturation in the chick embryonic heart. The phenotype observed during experiments was developmental regression of conduction system maturation together with changes in...

5 Abstract: Aims: The main objective of this work was to analyze in detail the effects of acute temperature changes on the function of isolated chick embryonic heart in vitro in comparison with natural conditions in ovo. Methods: The effects of temperature change (34 řC, 37 řC and 40 řC - hypo-, normo- and hyperthermia, respectively) on calcium dynamics in four days old isolated chick hearts in vitro were investigated by high-speed calcium optical imaging. For comparison and validation of in vitro measurements, experiments were also performed in ovo using videomicroscopy. Artificial electrical stimulation experiments were performed in vitro and in ovo to uncover conduction limits of different heart segments. Results: We observed almost linear dependence of sinus frequency on temperature in our temperature range. Sinus frequency during hypothermia and hyperthermia in vitro and in ovo changed about 20% in comparison with normothermia. We observed no significant changes in amplitude of calcium transients during temperature change to hypothermia but hyperthermia caused a significant decrease in amplitude of calcium transients (atria 35%, ventricles 38%). We observed a wide spectrum of arrhythmias, which occurred spontaneously even during normothermia in vitro. Occurrence of arrhythmias in vitro significantly...

Group of specialized cells that form cardiac conduction system is responsible for generation and coordinated propagation of the electrical impulse in the heart. Changes in its development can be connected with arrhythmias; therefore, a good level of knowledge is necessary and relevant for basic science and clinical practice. For correct development of the conduction system are important genes coding gap junctions proteins, ion channels, transcription factors and other molecules involved in signaling cascades (endothelin, neuregulin). Development of conduction system is determined in addition to genetic factors also by epigenetics and environmental factors. This thesis with its individual papers on which it is based is addressing different aspects of conduction system development, which appears to be a complex process. Another feature which is linking all papers together, is the methodological approach enabling us to study function of the conduction system - optical mapping. In the first publication we studied by the means of in vitro organ culture the impact of work load without interfering hemodynamics on the conduction system maturation in the chick embryonic heart. The phenotype observed during experiments was developmental regression of conduction system maturation together with changes in...

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Role of Nkx2.5 on development and electrophysiology of the mouse heart Prague 2015 Bc. Peter Hámor ABSTRACT The objective of this thesis is to investigate the role of Nkx2.5 gene dosage on electrophysiology of the mouse heart in prenatal stage of its development, in which the physiological functions of the heart fail to function properly. The main goal of this work is to search for differences in conduction of electric impulses through the embryonic mouse heart according to their genotype. Special method of capturing the conduction of electric impulse through myocardium was used for this purpose, called optical mapping. Thanks to this method I was able to construct images and videos capturing transition of the impulse with marked beginning of the activation and its direction in the heart. These outputs, or optical maps, help to define anomalies and defects compared with a normal functioning heart. The thesis focuses on the expression of the transcription factor Nkx2.5 and regulatory components related with the correct formation and physiology of the heart until 9.5 days post coitum. Individuals in this developmental stage were optically mapped and compared according to their genotypes - homozygous non-mutant, heterozygote and homozygous mutant mouse embryos exhibited some degree of similarity, while other...

Large scale image mosaicing methods are in great demand among scientists who study different aspects of the seabed, and have been fostered by impressive advances in the capabilities of underwater robots in gathering optical data from the seafloor. Cost and weight constraints mean that lowcost Remotely operated vehicles (ROVs) usually have a very limited number of sensors. When a low-cost robot carries out a seafloor survey using a down-looking camera, it usually follows a predetermined trajectory that provides several non time-consecutive overlapping image pairs. Finding these pairs (a process known as topology estimation) is indispensable to obtaining globally consistent mosaics and accurate trajectory estimates, which are necessary for a global view of the surveyed area, especially when optical sensors are the only data source. This thesis presents a set of consistent methods aimed at creating large area image mosaics from optical data obtained during surveys with low-cost underwater vehicles. First, a global alignment method developed within a Feature-based image mosaicing (FIM) framework, where nonlinear minimisation is substituted by two linear steps, is discussed. Then, a simple four-point mosaic rectifying method is proposed to reduce distortions that might occur due to lens distortions, error accumulation and the difficulties of optical imaging in an underwater medium. The topology estimation problem is addressed by means of an augmented state and extended Kalman filter combined framework, aimed at minimising the total number of matching attempts and simultaneously obtaining the best possible trajectory. Potential image pairs are predicted by taking into account the uncertainty in the trajectory. The contribution of matching an image pair is investigated using information theory principles. Lastly, a different solution to the topology estimation problem is proposed in a bundle adjustment framework. Innovative aspects include the use of fast image similarity criterion combined with a Minimum spanning tree (MST) solution, to obtain a tentative topology. This topology is improved by attempting image matching with the pairs for which there is the most overlap evidence. Unlike previous approaches for large-area mosaicing, our framework is able to deal naturally with cases where time-consecutive images cannot be matched successfully, such as completely unordered sets. Finally, the efficiency of the proposed methods is discussed and a comparison made with other state-of-the-art approaches, using a series of challenging datasets in underwater scenarios / Els mètodes de generació de mosaics de gran escala gaudeixen d’una gran demanda entre els científcs que estudien els diferents aspectes del fons submarí, afavorida pels impressionants avenços en les capacitats dels robots submarins per a l’obtenció de dades ptiques del fons. El cost i el pes constitueixen restriccions que impliquen que els vehicles operats remotament disposin habitualment d’un nombre limitat de sensors. Quan un robot de baix cost duu a terme una exploració del fons submarí utilitzant una càmera apuntant cap al terreny, aquest segueix habitualment una trajectòria que dóna com a resultat diverses parelles d’imatges amb superposició de manera sequencial. Trobar aquestes parelles (estimació de la topologia) és una tasca indispensable per a l’obtenció de mosaics globalment consistents així com una estimació de trajectòria precisa, necessària per disposar d’una visió global de la regió explorada, especialment en el cas en què els sensors òptics constitueixen la única font de dades. Aquesta tesi presenta un conjunt de mètodes robustos destinats a la creació de mosaics d’àrees de grans dimensions a partir de dades òptiques (imatges) obtingudes durant exploracions realitzades amb vehicles submarins de baix cost. En primer lloc, es presenta un mètode d’alineament global desenvolupat en el context de la generació de mosaics basat en característiques 2D, substituint una minimització no lineal per dues etapes lineals. Així mateix, es proposa un mètode simple de rectificació de mosaics basat en quatre punts per tal de reduir les distorsions que poden aparèixer a causa de la distorsió de les lents, l’acumulació d’errors i les dificultats d’adquisició d’imatges en el medi submarí. El problema de l’estimació de la topologia s’aborda mitjanant la combinació d’un estat augmentat amb un altre de Kalman estès, amb l’objectiu de minimitzar el nombre total d’intents de cerca de correspondències i obtenir simultàniament la millor trajectòria possible. La predicció de les parelles d’imatges potencials té en compte la incertesa de la trajectòria, i la contribució de l’obtenció de correspondències per a un parell d’imatges s’estudia d’acord amb principis de la teoria de la informació. Així mateix, el problema de l’estimació de la topologia és abordat en el context d’un alineament global. Les innovacions inclouen l’ús d’un criteri ràpid per a determinació de la similitud entre imatges combinat amb una solució basada en arbres d’expansió mínima, per tal d’obtenir una topologia provisional. Aquesta topologia és millorada mitjançant l’intent de cerca de correspondències entre parelles d’imatges amb major probabilitat de superposició. Contràriament al que succeïa en solucions prèvies per a la construcció de mosaics de grans àrees, el nostre entorn de treball és capaç de tractar amb casos en què imatges consecutives en el temps no han pogut ser relacionades satisfactòriament, com és el cas de conjunts d’imatges totalment desordenats. Finalment, es discuteix l’eficiència del mètode proposat i es compara amb altres solucions de l’estat de l’art, utilitzant una sèrie de conjunts de dades complexos en escenaris subaquàtics.