Introduction: Inflammatory cardiomyopathy (DCMi) represents a non-familial form of dilated cardiomyopathy (DCM) and endomyocardial biopsy (EMB) is crucial for its diagnosis. Aims: To assess the prevalence of DCMi in patients with DCM of unclear origin, to evaluate the significance of serological tests for antibodies against infectious cardiotrophic agents and to analyze the effect of specific therapy guided by EMB results. Methods: EMB was performed in 56 subjects (mean age 52 ± 10 years) with DCM of unclear etiology and left ventricular (LV) ejection fraction (EF) < 40% with a history of heart failure less than 1 year. EMB samples were analyzed by immunohistochemistry, polymerase chain reaction (PCR) and electron microscopy. Results: Immunohistochemical examination revealed myocardial inflammation in 26 patients (46%), the PCR method detected genome of microbial agents in 32 patients (57%). Electron microscopy showed the presence of particles of microbial agents in 41 patients (73%). Serological blood tests found no IgM antibody positivity against any of the investigated microbial agents. Targeted antibiotic therapy in patients with evidence of Borrelia burgdorferi (Bb) genome in the EMB led to a reduction in LV size, improvement of LV EF and alleviate symptoms of heart failure. Conclusion: DCMi...

Distinct cellular level of the Ca2+ binding chaperone calreticulin (CRT) is essential for cardiac development and postnatal function. However, CRT is also a potential autoantigen eliciting formation of antibodies (Ab), whose role is not yet clarified. Immunization with CRT leads to cardiac injury, and overexpression of CRT in cardiomyocytes induces dilated cardiomyopathy (DCM) in experimental animals. Hence, we analysed levels of anti-CRT Ab and calreticulin in the sera of patients with idiopatic DCM and hypertrophic cardiomyopathy (HCM). ELISA and immunoblot using human recombinant CRT and Pepscan with synthetic, overlapping decapeptides of CRT were used to detect anti-CRT Ab. Significantly increased levels of anti-CRT Ab of IgA (P<0.001) and IgG (P<0.05) isotypes were found in patients with both DCM (12/34 seropositive for IgA, 7/34 for IgG) and HCM (13/38 seropositive for IgA, 11/38 for IgG) when compared with controls (2/79 for IgA, 1/79 for IgG). Titration analysis in seropositive DCM and HCM patients documented anti-CRT Ab detected at 1/1600 dilution for IgG and 1/800 for IgA (and IgA1) and at least at 1/200 dilution for IgA2, IgG1, IgG2 and IgG3. Pepscan identified several immunogenic CRT epitopes: EVKIDNSQVESGSLED, IDDPTDSKPE, DKAPEHIPDPDA and RKEEEEAEDKEDDAEDKDEDEEDE recognised by IgA and...

Introduction: Inflammatory cardiomyopathy (DCMi) represents a non-familial form of dilated cardiomyopathy (DCM) and endomyocardial biopsy (EMB) is crucial for its diagnosis. Aims: To assess the prevalence of DCMi in patients with DCM of unclear origin, to evaluate the significance of serological tests for antibodies against infectious cardiotrophic agents and to analyze the effect of specific therapy guided by EMB results. Methods: EMB was performed in 56 subjects (mean age 52 ± 10 years) with DCM of unclear etiology and left ventricular (LV) ejection fraction (EF) < 40% with a history of heart failure less than 1 year. EMB samples were analyzed by immunohistochemistry, polymerase chain reaction (PCR) and electron microscopy. Results: Immunohistochemical examination revealed myocardial inflammation in 26 patients (46%), the PCR method detected genome of microbial agents in 32 patients (57%). Electron microscopy showed the presence of particles of microbial agents in 41 patients (73%). Serological blood tests found no IgM antibody positivity against any of the investigated microbial agents. Targeted antibiotic therapy in patients with evidence of Borrelia burgdorferi (Bb) genome in the EMB led to a reduction in LV size, improvement of LV EF and alleviate symptoms of heart failure. Conclusion: DCMi...

Dilated cardiomyopathy (DCM) is the second leading cause of heart failure. The pathophysiology in DCM is still poorly understood, partly because of currently limited research tools. We investigated whether cardiovascular magnetic resonance (CMR), using novel imaging techniques, could be used for in vivo assessment of some key pathophysiological mechanisms related to DCM. In addition, we evaluated whether the pathological findings on CMR would predict clinically relevant functional and morphological improvement of the left ventricular (LV) function - the LV reverse remodeling (LVRR). CMR together with endomyocardial biopsy, echocardiography, cardiopulmonary exercise testing and a thorough assessment of cardiac biomarkers was performed in 44 patients with new-onset DCM (<6 months of duration). The imaging was repeated after 12 months of clinical follow-up. Endomyocardial biopsy revealed myocardial inflammation in 34 % of the patients. LVRR at 12 months occurred in 45 % of the patients. Presence of late gadolinium enhancement (LGE) in the left ventricle was a sensitive but unspecific sign of myocardial inflammation because it was also a feature of hemodynamic stress related to the heart failure. The baseline extent of LGE was an independent predictor of future LVRR and also a predictor of adverse clinical...

Dilated cardiomyopathy (DCM) is the second leading cause of heart failure. The pathophysiology in DCM is still poorly understood, partly because of currently limited research tools. We investigated whether cardiovascular magnetic resonance (CMR), using novel imaging techniques, could be used for in vivo assessment of some key pathophysiological mechanisms related to DCM. In addition, we evaluated whether the pathological findings on CMR would predict clinically relevant functional and morphological improvement of the left ventricular (LV) function - the LV reverse remodeling (LVRR). CMR together with endomyocardial biopsy, echocardiography, cardiopulmonary exercise testing and a thorough assessment of cardiac biomarkers was performed in 44 patients with new-onset DCM (<6 months of duration). The imaging was repeated after 12 months of clinical follow-up. Endomyocardial biopsy revealed myocardial inflammation in 34 % of the patients. LVRR at 12 months occurred in 45 % of the patients. Presence of late gadolinium enhancement (LGE) in the left ventricle was a sensitive but unspecific sign of myocardial inflammation because it was also a feature of hemodynamic stress related to the heart failure. The baseline extent of LGE was an independent predictor of future LVRR and also a predictor of adverse clinical...

Heart transplantation is performed in patients with end-stage heart failure, in whom all other methods of treatment failed. The most common causes of end-stage heart failure are dilated cardiomyopathy and coronary artery disease. The destiny of these patients is highly variable. Prediction of long term survival in patients after heart transplantation is not satisfactory and up to now has not been found reliable marker. Most of the patients die after heart transplantation due to cardiovascular disease. This thesis is focused on molecular genetics and statistical analysis of four single nucleotide polymorphisms, namely rs17817449 (16q12.2, FTO gene), rs2943634 (2q36.3; intergenic region), rs6922269 (6q25.1; MTHFD1L gene), and rs10757274 (9p21.3; intergenic region). According to genome wide association studies are these SNPs assosiated with cardiovascular diseases. We genotyped DNA samples of 364 heart donors and 364 heart recipients. The results were statistically compared (using OR and Pearson's χ2 test) with the control group, which consisted of samples of individuals from the general population MONICA study. We examined the genotype in patients whose hearts failed due to dilated cardiomyopathy or coronary artery disease and then in patients with cardiac allograft vasculopathy. Furthermore, we focused on...

Sudden death in sport is a tragic event mediated primarily through the media. This thesis deals with sudden cardiac death from non-traumatic causes. The first section provides the overview of the issue, I mentioned specific cases of sudden death and the most common cause of sudden cardiac death among young and old athletes. The thesis focuses on the possibilities of preventive measures. In the second part, as part of my research, preparedness of Czech stadiums is evaluated situations of sudden cardiac death. In the conclusion section, I present the results of a questionnaire survey among representatives of clubs and stadiums and change proposals. The aim of the research is to find out how hockey and football stadiums are prepared for unpredictable situations of sudden cardiac death of athletes. Keywords: Sudden death, sport, sports stadiums, circulation disorder, hypertrophic cardiomyopathy, AED, resuscitation, EKG, prevention

Rare diseases (RD) are a heterogeneous group of diseases that affect about 5% of the world population. RDs represent more than 7.000 different phenotypes and many of them are genetically determined. RDs provide unique biological models for understanding the basic principles of molecular and cellular organization and function of human tissues and organs. Results of studies focused at pathogenesis of RDs are often used to diagnose and treat the affected patients. Significant progress in molecular genetic techniques, specifically the use of the next generation sequencing (NGS) in clinical practice, substantially facilitated and improved efficiency of RD laboratory diagnostics. Moreover, these novel testing algorithms identified the previously unknown molecular causes of many RDs. This thesis demonstrates the utility of NGS techniques and bioinformatics processing of obtained data in studies aimed at understanding molecular basis of selected RDs. These methods led to identification and characterization of causative pathogenic variants in the NDUFAF6 and PLD1 genes among patients affected by the Acadian variant of Fanconi disease and patients with a rare congenital heart defect, respectively. This approach was further used to analyze exomes of a large cohort of patients with different types of...

Rare diseases represent a clinically and genetically heterogeneous group of diseases affecting various organs and presenting at different ages. Identification and functional characterization of genetic defects causing individual rare diseases represent unique opportunity to understand biological functions of human genes and gene products as well as to basic pathogenetic mechanisms of individual diseases. This knowledge is prerequisite for their effective diagnosis, specific treatment and prevention and it also opens up an avenue for better understanding of complex diseases. My thesis documents basic conceptual and methodological developments of biochemical genetics, functional cloning, genetic mapping, positional cloning, DNA microarrays and genomic sequencing, which have provided a universal framework for effective characterization of the genetic architecture of almost all human diseases. This conceptual and technological developments are demonstrated on several cases of rare genetic diseases - adenylosuccinate lyase deficiency, mucopolysacharidosis type IIIC, Rotor syndrome, deficiency of ATP synthase, neuronal ceroid lipofuscinosis, GAPO syndrome and X -linked restrictive cardiomyopathy, which genetic and molecular basis I have helped to elucidate.

Introduction: Diagnosing inborn metabolic diseases, as a large subgroup of rare diseases, due to their rarity and wide variety of clinical manifestations, can be demanding and often prolonged. Objective: The aim of this work is, with the regard to clinical, biochemical and genetical aspects of selected rare diseases, to contribute to their rapid detection, widen the features of the natural course of the disease and contribute to their preventability. Material: This work includes cohort studies of patiens with cardiac manifestations in mitochondrial diseases, namely a group of 48 patients with TMEM70 protein deficiency, a group of 4 patients with Barth syndrome and individual cases of rare mitochondrial cardiomyopathies, thimidine kinase 2 deficiency and alanyl tRNA synthetase 2 deficiency. By determining the frequency, severity and type of heart disease, the phenotype was expanded, and the design of a therapeutic algorithm then made a positive impact on the prognosis of these patients. The work is also focused on the role of cardiac disease in the differential diagnosis of other genetically determined rare diseases, Marfan's syndrome and especially Pompe disease, where the emphasis is on early diagnosis, mainly due to the existence of an effective therapy. Focus on a broader differential diagnosis...