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The role of cold acclimation in ischemicko-reperfusion injury
Marvanová, Aneta ; Žurmanová, Jitka (advisor) ; Čížková, Dana (referee) ; Králová Lesná, Ivana (referee)
Cardiovascular disease, including ischemic heart disease, its acute form of myocardial infarction and subsequent heart failure, is still the most common cause of death in the world. For decades, scientists have been aiming to find clinically usable cardioprotective interventions that effectively prevented this disease or reduced the consequences of this disease and improved the quality of life of patients. It is known that a healthy lifestyle, which includes long-term hardening, contributes to heart protection. The beneficial effects of hardening on personality have been known for two hundred years, but no one before us has investigated the effect of a cold environment on the magnitude of ischemia-reperfusion (I/R) damage 10 years ago, we introduced a model of mild cold exposure (8 ± 1 řC) (MCE) which was protective on the size of the myocardial infarction and at the same time no negative side effects such as hypertension and myocardial hypertrophy were demonstrated. Our aim was to investigate the mechanism of cardioprotection induced by MCE. We asked the following questions: What is the metabolic profile of rats exposed to MCE? What is the effect of MCE on the magnitude of I/R damage in the short and long term? When does brown adipose tissue mature during MCE? Which signalling pathways are...
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Detection of nestin in the developing muscle and injury induced in vivo using a model of skeletal muscle lesion
Pechancová, Adéla ; Němečková, Ivana (advisor) ; Čížková, Dana (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Author: Adéla Pechancová Title of bachelor's thesis: Detection of nestin during development and injury with the use of in vivo model of skeletal muscle lesion Supervisor: RNDr. Ivana Němečková, PhD. Introduction and the aim of the study: Regeneration of tissues and organs is recently very actual topic. Regenerative medicine is coming to the foreground, it is a medical field focused on reparation of tissues. Intermediate filament nestin, which is considered to be a marker of stem cells and progenitor cells especially in nerve tissue, was also detected in immature elements of muscle tissue. The aim of this study was to prove expression of nestin during development and injury of skeletal muscle after injection of cardiotoxin and it's following regeneration using immunohistochemical methods. Methods: In the experimental part we were detecting nestin on paraffin embedded sections using mainly indirect immunohistochemical methods (LSAB, EnVision FLEX kit). The goal for the paraffin embedded section of mouse embryo was to capture the development of skeletal muscle. For the paraffin embedded section of leg muscle injured by cardiotoxin, the goal was to capture the amount of damage of the skeletal muscle and it's following ability to...
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Morphological and functional changes in microvasculature and endomysium in human atrial myocardium with atrial fibrillation
Smorodinova, Natalia ; Kučera, Tomáš (advisor) ; Čížková, Dana (referee) ; Čížková, Kateřina (referee)
This doctorial thesis was focused on morphological and functional changes in microvasculature and endomysium in human atrial myocardium with atrial fibrillation. Atrial fibrillation (AF) is one of the most common arrhythmias in the clinical practice and it is associated with an increase in mortality risk that is strongly related with old age. Its pathogenesis is still not sufficiently explored. One of the generally recognized factors contributing to the initiation and maintenance of atrial fibrillation is structural remodeling of the myocardium. Structural remodeling is reflected by changes that affect both atrial cardiomyocytes as well as endomysium. We analyzed atrial biopsies obtained from patients undergoing bypass or mitral valve surgery. The patients had a regular sinus rhythm or were suffering from AF. Immunohistochemistry was used to visualize collagen I, collagen III, elastin, desmin, smooth muscle actin and VEGF in the atrial samples.To detect capillaries UEA-lectin was used. For detection of different types of immune cells the following markers were detected immunohistochemically: CD45 as a pan- leukocyte marker, CD3 for T-lymphocytes, CD68 for monocyte/macrophages, mast cell tryptase for mast cells and DC-SIGN for immature dendritic cells. Our results document that in patients...
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Cryopreservation Methods in Manufacturing of Advanced Therapy Medicinal Products
Jandová, Miroslava ; Čížková, Dana (advisor) ; Stříbrná, Lucie (referee) ; Dubový, Petr (referee)
This dissertation deals with application of cryopreservation methods in the production and transport of advanced therapy medicinal products (ATMPs), including temporary storage in the Hospital cryostorage facility prior its clinical use. This work kept a strict adherence to European Union regulations and harmonized national legal norms. The first section shows the method validation regarding the collection and cryopreservation of starting material - peripheral blood mononuclear cells for the production of registered ATMPs using chimeric antigen receptor of T-lymphocytes (CAR-T). As a practical output from this endeavor, we obtained the approval for the collection, processing, and distribution of mononuclear cells for ATMP manufacture, as well as an export license to foreign facilities where the final production of genetically modified ATMPs takes place. The second part of this work presents the results of collection of starting material for ATMP production - bone marrow for manufacturing of human mesenchymal stromal cells (hMSCs)-based investigational ATMP and explains the development of the cryopreservation protocol for this investigational product under the conditions of good manufacturing practice (GMP) as an experimental part of the clinical trial EUDRA CT No. 2016-000926-21. The protocol was...
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Histological Evaluation of Experimental Models of Liver Diseases
Malečková, Anna ; Tonar, Zbyněk (advisor) ; Kachlík, David (referee) ; Čížková, Dana (referee)
Animal models are widely used for research of liver diseases pathogenesis and progression and for development of new treatment strategies in hepatology. The dissertation thesis focuses on large animal model, specifically swine. The use of animals, which are anatomically and physiologically close to humans, allows us to bridge the gap between the experimental and human medicine. Histopathological analysis of the liver biopsies is still a fundamental part of liver disease diagnosis and therefore, it is also a part of the experimental design of the studies using the porcine liver. Our aim was to apply qualitative and quantitative histological methods of evaluation on porcine liver and to assess their usability in experimental medicine. The quantitative methods included automated image analysis as well as stereological methods, which guaranteed high reproducibility and comparability of the experimental results. The dissertation thesis is based on 10 manuscripts. Three of them are published reviews associated with the main topic of the thesis. Seven original manuscripts resulted from six experimental studies - their six conclusions are listed bellow: Conclusion 1: We developed an open-source software QuantAn for quantification of microvessels visualized by 3D imaging methods, such as computed tomography...
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The role of mitochondrial genome in cardioprotection induced by the adaptation to chronic hypoxia
Nedvědová, Iveta ; Žurmanová, Jitka (advisor) ; Polák, Jan (referee) ; Čížková, Dana (referee)
Cardiovascular intervention studies are a very important issue given that the ischaemic heart disease is one of the main mortality and morbidity causes in the Western world. Cardioprotection is mediated through a variety of signalling pathways in the cell that may directly or indirectly affect energy metabolism and mitochondria. Ischaemia-reperfusion injury of the heart significantly affect mitochondrial function revealing a potential therapeutic target. The role of mitochondria in the myocardium is not only in the field of energy homeostasis, but also in mediating the cellular response to reduced oxygen supply and in apoptosis regulation. This thesis aims to elucidate the response of the hypertrophied heart of the spontaneously hypertensive rat (SHR) and the derived conplastic strain with mitochondrial genome of normotensive Brown Norway (SHR-mtBN ) to the cardioprotective regime of adaptation to chronic normobaric hypoxia (CNH, Fi 0.1). The adaptive changes were studied at the cellular, protein and gene levels using Real-time RT-PCR, Biomark Chip Analysis, Western Blot, spectrophotometric measurements of enzyme activity and quantitative immunofluorescence analyses. The present thesis was based on a different cardioprotective phenotype between SHR and SHR-mtBN strains, i.e. a significantly smaller...
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Invalid termination of the employment relationship by the employer
Čížková, Dana ; Drápal, Ljubomír (advisor) ; Morávek, Jakub (referee)
English Abstract Invalid termination of the employment relationship by the employer The topic of this thesis is the invalid termination of an employment relationship by the employer. The aim of the thesis is to conduct a comprehensive analysis of this point in labor law within the context of Act No. 89/2012 Coll., the Civil Code, which plays a major role in this issue as it pertains to subsidiary use in labor law. The Civil Code has brought a new understanding of legal defects of juridical acts that makes the juridical act either invalid or putative. It is just the putative juridical act that is a new institute implemented by the Civil Code. The relationship of the Labor Code to the Civil Code, including a historical excursion, is explained in the first part of the thesis. Furthermore, the thesis deals not only with the definition of the concepts of invalid and putative juridical acts in the context of labor law, but also with the development of these institutes in labor law. The main part of the thesis is devoted to the description and analysis of the individual cases of invalid or putative termination of an employment relationship by the employer, both on a general level and in specific cases using extensive case law. The thesis also deals with the individual consequences of invalid termination of the...
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Decaying fungi on selected sites in protected landscape area Karlštejn
Tesárková, Monika ; Pešková, Vítězslava (advisor) ; Čížková, Dana (referee)
The thesis aims to evaluate the state of health of tree species growing in Karlštejn National Nature Reserve, with the main focus on ligniperdous fungi found in this location.
Data were gathered in two experimental sites near Hostím and Bubovice villages, situated in the protected landscape area of Bohemian Karst (Český Kras). Tree species were inventoried in both designated sites and list of individual species of wood decaying fungi was put together. Furthermore, the state of health of tree species was evaluated according to crown defoliation and abiotic damage.
The thesis also briefly describes the history of Bohemian Karst Protected Landscape Area and other related themes climatic and soil conditions, including geological and geomorphological characteristics of this region that significantly influence the natural character of the territory, including occurrence of ligniperdous fungi.
The examination and subsequent controls of the selected sites took place once a month in period lasting from April 2015 to November 2016, revealed that only common species of wood decay fungi are found in the area of Karlštejn National Nature Reserve, which is part of Bohemian Karst Protected Landscape Area. These fungi are commonly present in the territory of the Czech Republic, therefore no unusual or new species of wood rotting fungi were found. The Korovitka (Diatrype) is the most widespread decaying fungi in both experimental plots, but the only one found fungal patogen Sphaeropsis sapinea, which in recent years has spread further and causes death and drying of pine trees has an economic importance for this area. Due to the good health of trees were found no major differences in the defoliation of trees, up to the aforementioned black pine attacked by fungi Sphaeropsis sapinea. There was observed defoliation from 45 to 100 %. There was no significant damage abiotic species found in the areas under examination.
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Health condition of trees in the arboretum Bukovina
Hochová, Lenka ; Čížková, Dana (advisor) ; Šrůtka, Petr (referee)
Diploma thesis Health condition of trees in the arboretum Bukovina explains the damage of trees, tree diseases, impacts of deseases in its theoretical part. It also includes a brief summary of biotic and abiotic factors that cause damage. The Manionova stress theory, which clearly indicates the gradual effect of stressors on the species, is described. The second chapter is focused on the ways how is the health condition of trees assessed. The typical rating scale is also mentioned. The methodics is focused on finding damage of trees or other anomalies with subsequent determination of the method of damage in the area of the arboretum. Location arboretum in the area Hruboskalsko rock town gives the possibility to find such damages that are typical for dense, mature stands. The first trees were planted in arboretum in 1860, so it is expected that on this old trees many diseases are evident. I focused on introduced tree species that are typical for this arboretum, it is also mentioned visible damage of native woods. The result of the work is to evaluate the status of selected species and the end of the thesis show the table with the detected results. At the end of the discussion the results are confronted and some recommendations are proposed.
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