Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

No description available.

<p>Disruption of coordinated impulse propagation in the heart as a result of fibrosis or myocardial infarction can create an asynchronous substrate with poor conduction and impaired contractility. This can ultimately lead to cardiac failure and make the heart more vulnerable to life-threatening arrhythmias and sudden cardiac death. The transplantation of exogenous cells into the diseased myocardium, "cardiac cell therapy," has been proposed as a treatment option to improve compromised cardiac function. Clinical trials of stem cell-based cardiac therapy have shown promising results, but also raised concerns about our inability to predict or control the fate of implanted cells and the electrical consequences of their interactions with host cardiomyocytes. Alternatively, genetically engineered somatic cells could be implanted to selectively and safely modify the cardiac electrical substrate, but their unexcitable nature makes them incapable of electrically repairing large conduction defects. The objective of this thesis was thus to develop a methodology to generate actively conducting excitable cells from an unexcitable somatic cell source and to demonstrate their utility for studies of basic electrophysiology and cardiac cell therapy.</p><p>First, based on the principles of cardiac action potential propagation, we applied genetic engineering techniques to convert human unexcitable cells (HEK-293) into an autonomous source of excitable and conducting cells by the stable forced expression of only three genes encoding an inward rectifier potassium (Kir2.1), a fast sodium (Na_v1.5), and a gap junction (Cx43) channel. Systematic pharmacological and electrical pacing studies in these cells revealed the individual contributions of each expressed channel to action potential shape and propagation speed. Conduction slowing and instability of induced arrhythmic activity was shown to be governed by specific mechanisms of I_Na inhibition by TTX, lidocaine, or flecainide. Furthermore, expression of the Na_v1.5 A1924T mutant sodium channel or Ca_v3.3 T-type calcium channel was utilized to study the specific roles of these channels in action potential conduction and demonstrate that genetic modifications of the engineered excitable cells in this platform allow quantitative correlations between single-cell patch clamp data and tissue-level function.</p><p>We further performed proof-of-concept experiments to show that networks of biosynthetic excitable cells can successfully repair large conduction defects within primary excitable tissue cultures. Specifically, genetically engineered excitable cells supported active action potential propagation between neonatal rat ventricular myocytes (NRVMs) separated by at least 2.5 cm in 2-dimensional and 1.3 cm in 3-dimensional cocultures. Using elastic films with micropatterned zig-zag NRVM networks that mimicked the tortuous conduction patterns observed in cardiac fibrosis, we showed that electrical resynchronization of cardiomyocyte activation by application of engineered excitable cells improved transverse conduction by 370% and increased cardiac twitch force amplitude by 64%. This demonstrated that despite being noncontractile, engineered excitable cells could potentially improve both the electrical and mechanical function of diseased myocardial tissue. </p><p>Lastly, we investigated how activation and repolarization gradients at the interface between cardiomyocytes and other excitable cells influence the vulnerability to conduction block. Microscopic optical mapping of action potential propagation was used to quantify dispersion of repolarization (DOR) in micropatterned heterocellular strands in which either well-coupled or poorly-coupled engineered excitable cells with a short action potential duration (APD), seamlessly interfaced with NRVMs that had a significantly longer APD. The resulting electrical gradients originating from the underlying heterogeneity in intercellular coupling and APD dispersion were further manipulated by the application of barium chloride (BaCl2) to selectively prolong APD in the engineered cells. We measured how the parameters of DOR affected the vulnerable time window (VW) of conduction block and found a strong linear correlation between the size of the repolarization gradient and VW. Reduction of DOR by BaCl2 significantly reduced VW and showed that VW correlated directly with dispersion height but not width. Conversely, at larger DOR, VW was inversely correlated with the dispersion width but independent of the dispersion height. In addition, despite their similar APDs, poorly-coupled excitable cells were found to significantly increase the maximum repolarization gradient and VW compared to well-coupled excitable cells, but only at larger DOR.</p><p>In summary, this thesis presents the novel concept of genetically engineering membrane excitability and impulse conduction in previously unexcitable somatic cells. This biosynthetic excitable cell platform is expected to enable studies of ion channel function in a reproducible tissue-level setting, promote the integration of theoretical and experimental studies of action potential propagation, and stimulate the development of novel gene and cell-based therapies for myocardial infarction and cardiac arrhythmias.</p> / Dissertation

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Abstract The quantitation of the variability in cardiovascular signals provides information about the autonomic neural regulation of the heart and the circulatory system. Several factors have an indirect effect on these signals as well as artifacts and several types of noise are contained in the recorded signal. The dynamics of RR and QT interval time series have also been analyzed in order to predict a risk of adverse cardiac events and to diagnose them. An ambulatory measurement setting is an important and demanding condition for the recording and analysis of these signals. Sophisticated and robust signal analysis schemes are thus increasingly needed. In this thesis, essential points related to ambulatory data acquisition and analysis of cardiovascular signals are discussed including the accuracy and reproducibility of the variability measurement. The origin of artifacts in RR interval time series is discussed, and consequently their effects and possible correction procedures are concidered. The time series including intervals differing from a normal sinus rhythm which sometimes carry important information, but may not be as such suitable for an analysis performed by all approaches. A significant variation in the results in either intra- or intersubject analysis is unavoidable and should be kept in mind when interpreting the results. In addition to heart rate variability (HRV) measurement using RR intervals, the dy- namics of ventricular repolarization duration (VRD) is considered using the invasively obtained action potential duration (APD) and different estimates for a QT interval taken from a surface electrocardiogram (ECG). Estimating the low quantity of the VRD vari- ability involves obviously potential errors and more strict requirements. In this study, the accuracy of VRD measurement was improved by a better time resolution obtained through interpolating the ECG. Furthermore, RTmax interval was chosen as the best QT interval estimate using simulated noise tests. A computer program was developed for the time interval measurement from ambulatory ECGs. This thesis reviews the most commonly used analysis methods for cardiovascular vari- ability signals including time and frequency domain approaches. The estimation of the power spectrum is presented on the approach using an autoregressive model (AR) of time series, and a method for estimating the powers and the spectra of components is also presented. Time-frequency and time-variant spectral analysis schemes with applica- tions to HRV analysis are presented. As a novel approach, wavelet and wavelet packet transforms and the theory of signal denoising with several principles for the threshold selection is examined. The wavelet packet based noise removal approach made use of an optimized signal decomposition scheme called best tree structure. Wavelet and wavelet packet transforms are further used to test their effciency in removing simulated noise from the ECG. The power spectrum analysis is examined by means of wavelet transforms, which are then applied to estimate the nonstationary RR interval variability. Chaotic modelling is discussed with important questions related to HRV analysis.ciency in removing simulated noise from the ECG. The power spectrum analysis is examined by means of wavelet transforms, which are then applied to estimate the nonstationary RR interval variability. Chaotic modelling is discussed with important questions related to HRV analysis.

Abstract Macrophages found in the tumor microenvironment play a crucial role in initiating an immunosuppressive tumor microenvironment that negatively impacts immunotherapy efficacy and aids tumor progression and metastasis. Constituting the most abundant immune cell in tumor microenvironment (TME), tumor associated macrophages (TAMs) have emerged as an attractive approach for anti-cancer therapy. However, two major challenges need to be overcome for successfully utilizing macrophages for immunotherapy. First, tumors repolarize the TAMs predominantly to M2 tumor-aiding phenotype by secreting various immunosuppressive cytokines. Second, cancer cells overexpress a membrane protein CD47 that interacts with signal-regulating protein alpha (SIRPalpha) expressed on macrophages. This crosstalk provides a downregulatory signal in the form of activation of SHP1/2 that inhibits cancer cell phagocytosis, and CD47, therefore, functions as a “don’t-eat-me” signal. We rationalized that these challenges can be overcome by engineering a nanoparticle system that can deliver a rationale combination of immunomodulatory agents to the TAMs that can both repolarize the M2 macrophages to M1 phenotype efficiently and concurrently block CD47-SIRPalpha interactions by inhibiting SHP2 signaling. Herein, we designed a lipid nanoparticle (LNP) system loaded with amphiphilic R848-cholesterol in its hydrophobic lipid bilayer, while SHP099 gets encapsulated in the hydrophilic core. Our previous studies have shown that the conjugation of cholesterol to the inhibitor stabilizes the lipid bilayer at a high inhibitor concentration. The LNPs showed high optimal drug loading, size, and stability. In vitro studies showed that the M2 macrophages treated with the LNPs system repolarized to M1 phenotype and expressed co-stimulatory molecules while having enhanced phagocytic potential. In vivo efficacy studies in 4T1 tumor-bearing mice showed that LNPs exhibit superior anti-tumor efficacy compared to other treatments. We evaluated the effect of MARCO-targeted LPNs by the conjugating anti-MARCO antibody on the LPN surface. However, no comparable difference in treatment efficacy was observed between the targeted MARCO-LNPs and the non-targeted LNPs. These results demonstrate that the MARCO targeting system designed in this study is largely ineffective in the targeted delivery of its drug cargo specifically to TAMs. Thus, the lipid nanoparticle-mediated co-delivery of a rational combination of TLR7/8 agonist and SHP2 inhibitor in the TAMs increases M2 to M1 repolarization and phagocytosis potential of macrophages. Recommended Citation Malik, V., Ramesh, A. and Kulkarni, A.A. (2021), TLR7/8 Agonist and SHP2 Inhibitor Loaded Nanoparticle Enhances Macrophage Immunotherapy Efficacy. Adv. Therap., 4: 2100086. https://doi.org/10.1002/adtp.202100086

Le tamoxifène, un modulateur sélectif des récepteurs oestrogéniques, est un médicament largement utilisé depuis plus de vingt ans pour le traitement et la prévention du cancer du sein. Plusieurs études ont rapporté que l’administration aiguë du tamoxifène pouvait réduire certains courants K+ cardiaques. Cette observation suggère que les femmes traitées de façon chronique avec le tamoxifène risquent d’avoir une prolongation de leur intervalle QT, favorisant ainsi le développement de torsades de pointes. Puisque in vivo, le tamoxifène est largement métabolisé et son effet est attribué à celui du 4hydroxy-tamoxifène (4OH-tamoxifène), nous avons d'abord vérifié si les effets du tamoxifène sur la repolarisation pouvaient être dus au 4OH-tamoxifène. À l'aide de la méthode de patch-clamp, nous avons étudié l’effet aigu du 4OH-tamoxifène sur les courants K+ présents au niveau ventriculaire chez la souris femelle. En premier lieu, nous avons démontré que les souris traitées avec le 4OH-tamoxifène présentaient une diminution des courants K+ comparativement aux souris intactes. Fait intéressant, le prétraitement des myocytes avec l’antagoniste des récepteurs oestrogéniques, le ICI 182,780, ou l’inhibiteur de la synthèse protéique, l'actinomycine D, n’a pas modifié les effets du 4OH-tamoxifène. Ces résultats suggéraient que les effets du 4OH-tamoxifène sur les courants potassiques ne soient pas liés à la transcription génomique et n’implique pas les récepteurs aux œstrogènes. Bien que l’administration aiguë du 4OH-tamoxifène diminue les courants K+ cardiaques, l’absence de troubles au niveau du rythme cardiaque chez les femmes traitées à long terme exclu la possibilité de conclure que le traitement chronique avec le tamoxifène augmente la durée de l’intervalle QT. L'accès à des souris femelles et des cobayes nous a permis de démontrer que contrairement au traitement en aigu, les courants et les canaux K+ cardiaques sont augmentés en chronique. Les oestrogènes associés à une diminution des courants K+ d’une part et nos résultats obtenus avec le tamoxifène d’autre part suggèrent qu’en bloquant les récepteurs oestrogéniques, le tamoxifène puisse prévenir les effets inhibiteurs des oestrogènes sur les courants K+. Cette association œstrogènes- tamoxifène- récepteurs oestrogéniques et courants K+ nous a encouragées à approfondir encore nos études et vérifier l’influence des hormones sexuelles féminines sur la repolarisation ventriculaire. Une troisième étude a été ainsi réalisée chez des souris femelles ovariectomisées et des souris déficientes en récepteurs oestrogéniques α ou β afin de vérifier le rôle des oestrogènes et des récepteurs oestrogéniques sur la repolarisation ventriculaire. Nos résultats ont révélé clairement que l’absence des oestrogènes entraîne une augmentation de la densité du courant K+ transitoire indépendant du Ca2+ (Ito) et de l’expression du canal Kv4.3 et ces effets sont médiés par les REα. Ces données soutiennent davantage notre conclusion que l’inhibition des récepteurs oestrogéniques est responsable de l’augmentation des courants/canaux K+ et suggèrent fortement qu’ils jouent un rôle dans la régulation de la repolarisation ventriculaire. Elles soulignent aussi l'importance de vérifier le statut hormonal des animaux utilisés pour des études touchant l'électrophysiologie cardiaque. Dans la dernière partie de cette thèse nous avons vérifié les effets de la grossesse et du système nerveux autonome sur les différents paramètres électrocardiographiques et plus particulièrement sur le rythme cardiaque chez la souris. Nos données ont montré que, comme chez la femme enceinte, la grossesse est associée à une augmentation du rythme cardiaque. De plus, l'augmentation des niveaux des hormones féminines pourrait affecter l’automatisme et l’activité électrique cardiaque. Ces différentes études ont augmenté les connaissances sur la régulation hormonale de l'électrophysiologie cardiaque et aideront aux avancements des recherches chez les femmes. / Tamoxifen is a selective estrogen receptor modulator widely used in the treatment and prevention of breast cancer from more than 20 years. Other studies have reported that acute exposure to tamoxifen can reduce cardiac K+ currents. However, in vivo tamoxifen is largely metabolized and most of its activity is attributable to its major metabolite, 4- hydroxytamoxifen (4OH-tamoxifen). In our first study, we investigated the acute effects of 4OH-tamoxifen on cardiac K+ currents in mice. Using the patch-clamp technique, we found that, as with tamoxifen, short-term exposure to 4OH-tamoxifen reduced K+ currents in the mouse ventricle even in the presence of the selective oestrogen receptor antagonist, ICI-182,780, or the inhibitor of RNA synthesis, actinomycin D. These results suggest an inhibition independent of the intracellular oestrogen receptor and the protein synthesis. However, women receiving long-term tamoxifen therapy do not experience cardiac arrhythmias although acute perfusion of tamoxifen has been shown to inhibit cardiac K+ currents. This observation suggests that chronic tamoxifen treatment does not negatively modulate cardiac K+ currents. Therefore, in order to investigate the absence of cardiac arrhythmias in women, we examined the effects of long term tamoxifen therapy associated with low level of estrogen on ventricular K+ currents and channels expressed in mouse and guinea pig heart. Female mice and guinea pigs were treated with placebo or tamoxifen pellets for 60 days. Our results show an increase of the densities of K+ currents and the expression of their channels. Conditions with high oestrogen levels are associated with reduced K+ currents both in the heart and the uterus. Thus, conceivably, tamoxifen might prevent the inhibitory effects of oestrogen on K+ channels by blocking the oestrogen receptors, which would explain the reported increase in K+ currents. These findings could contribute to explain the absence of cardiac arrhythmia with long-term tamoxifen therapy. These association estrogen- tamoxifen and estrogen receptor lead us to study the influence of estrogens and estrogen receptors on ventricular repolarization. Accordingly, we assessed the involvement of estrogens and ER on K+ currents in mouse using ovariectomised (OVX), ER-knockout (ERKOα) or ERβ-knockout (ERKOβ) female mice. These experiments show that the absence of estrogen is associated with an increase of the density of the Ca2+-independent transient outward, Ito, the mRNA and protein expression of Kv4.3. These results vi obtained with ERKO mice suggest that estrogens regulate ventricular repolarization and ER mediates this effect. The last part of this thesis was to determine whether pregnancy elicits a change in the heart rate in mice and whether pregnancy-related changes are due to the cardiac conduction system rather than a change in autonomic tone. Our results revealed that pregnancy accelerates the resting heart rate and the AV node conduction time in the presence and absence of autonomic nervous system input suggesting an intrinsic mechanism. Moreover, Hormonal changes that occur during pregnancy are likely to be involved in these adaptations of the heart to pregnancy. These studies provide a new insight to understand the hormonal regulation of cardiac electrophysiology.

Le tamoxifène, un modulateur sélectif des récepteurs oestrogéniques, est un médicament largement utilisé depuis plus de vingt ans pour le traitement et la prévention du cancer du sein. Plusieurs études ont rapporté que l’administration aiguë du tamoxifène pouvait réduire certains courants K+ cardiaques. Cette observation suggère que les femmes traitées de façon chronique avec le tamoxifène risquent d’avoir une prolongation de leur intervalle QT, favorisant ainsi le développement de torsades de pointes. Puisque in vivo, le tamoxifène est largement métabolisé et son effet est attribué à celui du 4hydroxy-tamoxifène (4OH-tamoxifène), nous avons d'abord vérifié si les effets du tamoxifène sur la repolarisation pouvaient être dus au 4OH-tamoxifène. À l'aide de la méthode de patch-clamp, nous avons étudié l’effet aigu du 4OH-tamoxifène sur les courants K+ présents au niveau ventriculaire chez la souris femelle. En premier lieu, nous avons démontré que les souris traitées avec le 4OH-tamoxifène présentaient une diminution des courants K+ comparativement aux souris intactes. Fait intéressant, le prétraitement des myocytes avec l’antagoniste des récepteurs oestrogéniques, le ICI 182,780, ou l’inhibiteur de la synthèse protéique, l'actinomycine D, n’a pas modifié les effets du 4OH-tamoxifène. Ces résultats suggéraient que les effets du 4OH-tamoxifène sur les courants potassiques ne soient pas liés à la transcription génomique et n’implique pas les récepteurs aux œstrogènes. Bien que l’administration aiguë du 4OH-tamoxifène diminue les courants K+ cardiaques, l’absence de troubles au niveau du rythme cardiaque chez les femmes traitées à long terme exclu la possibilité de conclure que le traitement chronique avec le tamoxifène augmente la durée de l’intervalle QT. L'accès à des souris femelles et des cobayes nous a permis de démontrer que contrairement au traitement en aigu, les courants et les canaux K+ cardiaques sont augmentés en chronique. Les oestrogènes associés à une diminution des courants K+ d’une part et nos résultats obtenus avec le tamoxifène d’autre part suggèrent qu’en bloquant les récepteurs oestrogéniques, le tamoxifène puisse prévenir les effets inhibiteurs des oestrogènes sur les courants K+. Cette association œstrogènes- tamoxifène- récepteurs oestrogéniques et courants K+ nous a encouragées à approfondir encore nos études et vérifier l’influence des hormones sexuelles féminines sur la repolarisation ventriculaire. Une troisième étude a été ainsi réalisée chez des souris femelles ovariectomisées et des souris déficientes en récepteurs oestrogéniques α ou β afin de vérifier le rôle des oestrogènes et des récepteurs oestrogéniques sur la repolarisation ventriculaire. Nos résultats ont révélé clairement que l’absence des oestrogènes entraîne une augmentation de la densité du courant K+ transitoire indépendant du Ca2+ (Ito) et de l’expression du canal Kv4.3 et ces effets sont médiés par les REα. Ces données soutiennent davantage notre conclusion que l’inhibition des récepteurs oestrogéniques est responsable de l’augmentation des courants/canaux K+ et suggèrent fortement qu’ils jouent un rôle dans la régulation de la repolarisation ventriculaire. Elles soulignent aussi l'importance de vérifier le statut hormonal des animaux utilisés pour des études touchant l'électrophysiologie cardiaque. Dans la dernière partie de cette thèse nous avons vérifié les effets de la grossesse et du système nerveux autonome sur les différents paramètres électrocardiographiques et plus particulièrement sur le rythme cardiaque chez la souris. Nos données ont montré que, comme chez la femme enceinte, la grossesse est associée à une augmentation du rythme cardiaque. De plus, l'augmentation des niveaux des hormones féminines pourrait affecter l’automatisme et l’activité électrique cardiaque. Ces différentes études ont augmenté les connaissances sur la régulation hormonale de l'électrophysiologie cardiaque et aideront aux avancements des recherches chez les femmes. / Tamoxifen is a selective estrogen receptor modulator widely used in the treatment and prevention of breast cancer from more than 20 years. Other studies have reported that acute exposure to tamoxifen can reduce cardiac K+ currents. However, in vivo tamoxifen is largely metabolized and most of its activity is attributable to its major metabolite, 4- hydroxytamoxifen (4OH-tamoxifen). In our first study, we investigated the acute effects of 4OH-tamoxifen on cardiac K+ currents in mice. Using the patch-clamp technique, we found that, as with tamoxifen, short-term exposure to 4OH-tamoxifen reduced K+ currents in the mouse ventricle even in the presence of the selective oestrogen receptor antagonist, ICI-182,780, or the inhibitor of RNA synthesis, actinomycin D. These results suggest an inhibition independent of the intracellular oestrogen receptor and the protein synthesis. However, women receiving long-term tamoxifen therapy do not experience cardiac arrhythmias although acute perfusion of tamoxifen has been shown to inhibit cardiac K+ currents. This observation suggests that chronic tamoxifen treatment does not negatively modulate cardiac K+ currents. Therefore, in order to investigate the absence of cardiac arrhythmias in women, we examined the effects of long term tamoxifen therapy associated with low level of estrogen on ventricular K+ currents and channels expressed in mouse and guinea pig heart. Female mice and guinea pigs were treated with placebo or tamoxifen pellets for 60 days. Our results show an increase of the densities of K+ currents and the expression of their channels. Conditions with high oestrogen levels are associated with reduced K+ currents both in the heart and the uterus. Thus, conceivably, tamoxifen might prevent the inhibitory effects of oestrogen on K+ channels by blocking the oestrogen receptors, which would explain the reported increase in K+ currents. These findings could contribute to explain the absence of cardiac arrhythmia with long-term tamoxifen therapy. These association estrogen- tamoxifen and estrogen receptor lead us to study the influence of estrogens and estrogen receptors on ventricular repolarization. Accordingly, we assessed the involvement of estrogens and ER on K+ currents in mouse using ovariectomised (OVX), ER-knockout (ERKOα) or ERβ-knockout (ERKOβ) female mice. These experiments show that the absence of estrogen is associated with an increase of the density of the Ca2+-independent transient outward, Ito, the mRNA and protein expression of Kv4.3. These results vi obtained with ERKO mice suggest that estrogens regulate ventricular repolarization and ER mediates this effect. The last part of this thesis was to determine whether pregnancy elicits a change in the heart rate in mice and whether pregnancy-related changes are due to the cardiac conduction system rather than a change in autonomic tone. Our results revealed that pregnancy accelerates the resting heart rate and the AV node conduction time in the presence and absence of autonomic nervous system input suggesting an intrinsic mechanism. Moreover, Hormonal changes that occur during pregnancy are likely to be involved in these adaptations of the heart to pregnancy. These studies provide a new insight to understand the hormonal regulation of cardiac electrophysiology.

Abstract Based on experimental studies, heterogeneous ventricular repolarization has been suggested to predispose to the onset of life-threatening ventricular arrhythmias and subsequent studies in various patient populations have associated it with poor clinical outcome. The aim of this study was to assess the dynamics of ventricular repolarization from electrocardiogram (ECG) during a standard exercise stress test with techniques based on principal component analysis, and to investigate whether they would yield valuable prognostic information on cardiac mortality and sudden cardiac death. Exercise ECG recordings from 40 healthy men and women were analyzed in order to establish normal dynamics and rate-dependence for the spatial QRS/T angle and total cosine R to T (TCRT), which reflects the spatial deviation between the depolarization and repolarization wave-fronts. Subsequently, exercise ECG recordings from 20 patients with previous myocardial infarction (MI) were compared with 20 controls in order to assess the modulation of previous MI on the dynamics of TCRT. The prognostic value of abnormal dynamics and rate-dependence of TCRT and spatial QRS/T angle were assessed in 1,297 patients referred to a clinically indicated exercise stress test. The spatial QRS/T angle and TCRT were found to be significantly rate-dependent during both exercise and recovery periods as TCRT and the cosine of the spatial QRS/T angle decreased at higher heart rates (HR), indicating increase in the global heterogeneity of the ventricular repolarization. In the post-MI group, the dynamics of TCRT were impaired with a significantly lower rate-dependence in contrast to the healthy and control groups. In the study cohort, impaired rate-dependence of TCRT during both exercise and recovery periods was an independent predictor of cardiac and sudden cardiac death during the follow up of the study. Similarly, reduction of the area between the exercise and three-minute recovery TCRT/HR curves was associated with an increased risk of cardiac and sudden cardiac death. In conclusion, dynamics and rate-dependence of TCRT and spatial QRS/T angle are able to discriminate subjects with an increased risk of cardiac and sudden cardiac death. Reduced rate-dependence of TCRT during exercise and recovery periods, as well as decreased exercise-recovery hysteresis of TCRT, are significant predictors of cardiac and sudden cardiac death. / Tiivistelmä Sydämen kammioiden poikkeavan palautumisen on kokeellisissa tutkimuksissa havaittu altistavan kammioperäisille rytmihäiriöille. Lisäksi, useat tutkimukset erilaisissa potilasaineistoissa ovat osoittaneet sen olevan yhteydessä heikkoon kliiniseen ennusteeseen. Tämän työn tarkoituksena oli analysoida sydämen kammioiden repolarisaation dynamiikkaa sydänsähkökäyrästä (EKG) rasituskokeen aikana pääkomponenttianalyysiin perustuvilla menetelmillä sekä tutkia, ennustavatko uudet muuttujat sydänperäistä kuolemaa tai sydänperäistä äkkikuolemaa. Neljänkymmenen terveen miehen ja naisen rasitus-EKG:t analysoitiin depolarisaatio- ja repolarisaatioaaltojen välistä avaruudellista kulmaa kuvaavan ’total cosine R to T’:n (TCRT) sekä kolmiulotteisen QRS/T-kulman dynamiikan sekä sykeriippuvuuden normaaliarvojen määrittämiseksi. Lisäksi, vertailtiin 20 sydäninfarktin sairastaneen sekä 20 kontrollipotilaan rasitusmittauksia aiemman infarktin TCRT:n dynamiikkaan sekä sykeriippuvuuteen aiheuttaman modulaation määrittämiseksi. Muuttujien poikkeavan dynamiikan ennustearvoa testattiin 1297 rasitus-EKG-mittaukseen lähetteen saaneella potilaalla. Sekä TCRT että avaruudellisen QRS/T-kulman kosini olivat molemmat erittäin sykeriippuvaisia sekä rasituksessa että palautumisessa. Molemmat muuttujat pienenivät sykkeen (HR) kasvaessa, osoittaen kammioiden repolarisaation heterogeenisyyden kasvua korkeammilla syketaajuuksilla. Sydäninfarktin sairastaneilla potilailla muuttujien dynamiikka ja sykeriippuvuus olivat heikentyneitä. Kohorttiaineistossa heikentynyt sykeriippuvuus sekä rasituksen että palautumisen aikana ennustivat myöhempää sydänperäistä kuolemaa sekä sydänperäistä äkkikuolemaa itsenäisesti. Vastaavasti, rasituksen ja kolmen minuutin palautumisen aikaisten TCRT/HR kuvaajien väliin jäävän pinta-alan pienentyminen oli yhteydessä korkeampaan sydänperäisen kuoleman sekä sydänperäisen äkkikuoleman riskiin. Kammioiden repolarisaation dynamiikkaa kuvaavat muuttujat pystyvät erottelemaan potilaat, joilla on korkeampi riski kokea sydänperäinen kuolema tai sydänperäinen äkkikuolema. Poikkeavan alhainen sykeriippuvuus rasituksen ja palautumisen aikana sekä alentunut rasituksen ja palautumisen välinen hystereesi olivat merkittäviä sydänperäisen kuoleman sekä sydänperäisen äkkikuoleman ennustajia.

The thesis deals with possibilities of using wavelet transform in the field of surface electrocardiogram (ECG) signals denoising and ECG signals measuring. Several algorithms have been used to detect and estimate T-wave alternans (TWA), such as spectral method (SM), Poincaré Mapping (PM) or correlation method (CM). T-wave alternans, also called repolarization alternans, is a phenomenon appearing in the electrocardiogram as a consistent fluctuation in the repolarization morphology on every-other-beat basis. Electrical TWA has been recognized as a marker of electrical instability, and has been shown to be related with patients at increased risk for ventricular arrhytmias. Presence of TWA has been reported in a wide range of clinical and experimental situations including long QT syndrome, myocardial infarction, angina pectoris, acute ischemia, etc. Projected methods of detection TWA are realized in Matlab software, and they are experimentally verified on real ECG signals from the European ST-T Database.