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Study of interactions participating in the separation mechanism of chromatographic separation systems
Kalíková, Květa ; Tesařová, Eva (advisor) ; Pacáková, Věra (referee) ; Ševčík, Juraj (referee)
Charles University in Prague Faculty of Science Department of Physical and Macromolecular Chemistry Study of interactions participating in the separation mechanism of chromatographic separation systems The PhD Thesis Report Květa Kalíková Prague 2009 The PhD thesis was carried out within the PhD study during years 2005-2009 at the Department Physical and Macromolecular Chemistry, Faculty of Science, Charles University in Prague. Supervisor: Doc. RNDr. Eva Tesařová, CSc. 2 Content 1. Introduction 4 2. Aims of the thesis 4 3. Results and discussion 5 3.1 Characterization of separation system using LFER model (publication I) 5 3.2 Occurrence and behavior of system peaks in RP HPLC with solely aqueous mobile phases (publication II) 6 3.3 Examples of optimized separation systems and their application 8 3.3.1 Achiral separation systems 8 3.3.1.1 HPLC separation of estrogens (publication III) 8 3.3.1.2 HPLC method for separation of 1,4-benzodiazepines (publication in progress) 9 3.3.2 Chiral separation systems (publication IV) 10 3.3.2.1 Optimization and validation of HPLC method for enantioseparation of (±)-cloprostenol (publication V) 11 4. Conclusions 12 5. References 13 6. List of publications 14 7. List of presentations 15 8. Curriculum Vitae 17 3 1. Introduction High performance liquid chromatography (HPLC)...
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Heidegger's Concept of Experience
Ševčík, Jan ; Benyovszky, Ladislav (advisor) ; Němec, Václav (referee)
Heidegger's Concept of Experience Abstract The paper aims to explore one of the central concepts of Heidegger's late philosophy. The experience here, compared to the concept in the early period of his thinking, takes on new forms of relation to the world and to the concealment and shows some basic features that the work will follow. These features of experience are: totality, immediacy, passivity, and the transformation of whoever experiences. A proper understanding of Heidegger's concept of experience should provide an important clue to understanding his late philosophy and the way of thinking that promotes it. But it also has wider implications for today's thinking and for grasping not only prominent experiences, such as religious experience.
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Využití zemědělského půdního fondu v katastrálním území Orlovice
Ševčík, Jan
This bachelor thesis deals with an evolution and use of an agricultural land fund in a cadastre unit of Orlovice, based on studies of historical and present map bases analyses. The work focuses on comparison of historical and present states of the agricultural land fund, general information of the agricultural land fund and an evolution of land ownership relations. In the thesis, I suggested provisions which would contribute to land protection and helped to general improvement of the landscape character in the area.
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Characterization of separation systems for determination of enantiomers
Geryk, Radim ; Tesařová, Eva (advisor) ; Ševčík, Juraj (referee) ; Nováková, Lucie (referee)
(EN) The dissertation thesis is focused on the research and characterization of retention and enantiorecognition mechanisms of chiral stationary phases based on derivatized polysaccharides. The separation systems with a variety of modern stationary phases (both achiral and chiral) were characterized in detail to provide a comprehensive view of the interactions participating in the separation process. The study of the retention/separation behavior significantly facilitates the development and the optimization of new enantioselective methods for a wide variety of compounds. The work deals with the comparison of enantioselective performance of polysaccharide-based chiral stationary phases. The objectives are to show the differences of separation behavior among these chiral stationary phases, as they differ by the nature of the polysaccharide backbone (amylose versus cellulose), by binding of chiral polymer to silica support (coated versus immobilized stationary phase) and by the phenyl moiety in the reversed and normal phase HPLC. In both separation modes amylose-based chiral stationary phases exhibited higher enantioselectivity, especially for acidic and bifunctional analytes. Chiral stationary phases based on derivatized cellulose showed higher enantiodiscrimination potential for basic analytes....
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Study of population specific alterations of breast cancer predisposition genes in Czech Republic.
Judasová, Kristýna ; Ševčík, Jan (advisor) ; Holá, Dana (referee)
Breast cancer is the most frequent malignant disease in the female population worldvide. About 10 % of all cases are of hereditary origin. The inactivation of tumor suppressor gene BRCA1 is the main genetic predisposing factor in breast cancer in the Czech Republic. Primarily, BRCA1 participates in DNA double strand break repair. Depending on cell cycle phase, the damage is repaired by homologous recombination or non-homologous end joining. Alternative splicing variants of BRCA1 are frequently detected during the genetic screening of high risk patients. The clinical significance of these variants is unknown. Understanding of the nature of breast cancer genetics is the critical factor for early diagnosis. Based on earlier studies from the Institute of Biochemistry and Experimental Oncology 1st Faculty of Medicine Charles University, two alternative splicing variants which were repeatedly detected in patients, were chosen for functional analysis. The aim of this work is to investigate the impact of alternative splicing variants BRCA1Δ5 and BRCA1Δ10 on DNA double strand breaks repair. Particular variants were over- expressed in the cells of model system. Activity of homologous recombination (HR) and non-homologous end joining (NHEJ) was scored by in vitro DNA repair assay. The cellular localization of...
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Establishment and Development of European Scheme for Greenhouse Gas Emission Allowance Trading
Ševčík, Jakub ; Žákovská, Karolina (advisor) ; Damohorský, Milan (referee)
The aim of this thesis is to describe and subsequently analyze the establishment and the development of the European legal regime for greenhouse gas emission allowance trading. Methodology of the thesis is focused on capturing the processes of establishment and development of the emission trading regime as processes of rather dynamic nature, having their causes and consequences, and so is not only an isolated description of each of their respective stages. The dynamics of development is analyzed within each of the basic elements of the emission trading regime, taking due account of the relevant case law of the Court of Justice of the EU. Given the considerable complexity of the subject concerned, and therefore also the need for integration of the analysis into a certain perspective, a brief explanation of the phenomenon of climate change, including its economic implications, as well as of the international legal framework set up to tackle this issue forms a part of this thesis.
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The functional in vitro analysis of the BRCA1alternative splicing variants
Ševčík, Jan ; Kleibl, Zdeněk (advisor) ; Stopka, Tomáš (referee) ; Macůrek, Libor (referee)
BACKGROUND: The inactivation of the tumor suppressor gene BRCA1 is a predisposing factor for a breast/ovarian cancer development. Formation of cancer-specific alternative splicing variants with aberrant biological properties can represent additional mechanism decreasing the overall BRCA1 activity in DNA double strand break (DDSB) repair. In this study, we analyzed BRCA1 alternative splicing variants BRCA114-15 and 17-19 ascertained previously during the screening of high-risk breast cancer individuals. METHODS: We established a stable MCF-7 cell line-based model system for an in vitro analysis of BRCA1 variants. Using this system, we analyzed the impact of BRCA114-15 and 17-19 variants on DNA repair kinetics using comet assay and confocal immunomicroscopy. The capacity of DNA repair was assessed directly by an in vitro NHEJ assay and indirectly by a mitomycin C sensitivity test. The proliferation activities were determined by a clonogenic assay and growth curves. RESULTS: Overexpression of BRCA114-15 and 17-19 increases the endogenous level of DNA damage, slows down the DDSB repair, and decelerates the initial phase of radiation-induced foci formation and prolongs their persistence. Moreover, BRCA114-15 and 17-19 differentially influence the activity of HR and NHEJ and sensitivity of MCF-7 cells to ionizing...
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A DNA double-strand break repair and it's disorders with a relationship to the cancerogenesis
Komžák, Josef ; Ševčík, Jan (advisor) ; Vopálenský, Václav (referee)
The DNA-double strand break (DSB) repair has an essential importance for the genomic integrity maintenance. The main DSB repair pathways are homologous recombination (HR), non-homologous end joining (NHEJ) and single-strand annealing (SSA). The most important protein factors contributing to the maintenance of genomic integrity by direct participation in DSB repair are MRN, ATM, Rad51, BRCA1/2 and PALB2 in the case of HR; Ku70/80 DNA-PKcs, XRCC4 and DNA ligase IV in the case of NHEJ and Msh2-Msh3 and Rad1- Rad10 in the case of SSA. If mutated, these proteins can cause the inability to repair DNA lesions leading to a malignant transformation. The predominant phenotype manifestation of BRCA1/2 inactivation is the hereditary breast and/or ovarian cancer (HBOC). Mutations in ATM have been described as a cause of ataxia telangiectasia and inactivation of NBN gene (Nbs1 protein) causes the Nijmegen breakage syndrome. Other syndromes connected with defects in a DSB repair pathways are Fanconi anemia and Werner syndrome. Detail knowledge of DSB repair process is a mandatory for diagnostics and effective therapy of a number of malignances. An example of practical and clinically relevant utilization of current knowledge about the DSB repair process is the concept of a synthetic lethality as a specific therapy. This...
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The role of proto-oncogene crk in invasiveness
Tomášová, Lea ; Rösel, Daniel (advisor) ; Ševčík, Jan (referee)
Proto-oncogene Crk was identified as an oncogenic product of an avian retrovirus in 1988. It is an adaptor protein containing SH2 and SH3 binding domains. Thanks to these domains Crk facilitates protein-protein interactions and therefore plays a crucial role in signal transduction. Crk forms signal complexes with several proteins and hence impacts many cellular processes, among them cell migration, tumorigenezis and invasion of the surrounding tissues. The increased invasiveness allows the tumour cells to detach from the primary tumour and form metastasis which is very problematic feature of cancer. Overexpression of Crk was observed in several tumour tissues, it correlates with an aggressive and metastatic phenotype of the tumours. The subject of this thesis is to describe the mechanisms of how Crk can regulate cellular motility and invasiveness.
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