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Study of Glycine-NBDCl Derivate Stability
Koudelková, Lenka ; Kubíček, Vladimír (advisor) ; Lázníčková, Alice (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of biophysics and physical chemistry Candidate: Lenka Koudelková Consultant: Ing.Vladimír Kubíček, CSc. Title of Thesis: Study of glycine-NBDCl derivate stability In this thesis the stability of glycine-NBDCl derivative under three different conditions (heat+light, heat+dark, cool+dark) is studied by HPLC with fluorimetric detection. Ater being stabilized by hydrochloric acid the derivatives exhibit a better stability, but the concentrations found during several weeks are too scattered. In addition an alternative chromatographic conditions were proposed for the HPLC analysis. Acetonitrile with hydrogen phosphate buffer (pH 7,0) in ratio 70:30 on an Atlantis column gave symetric peaks with a short time of elution.
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Structural and regulatory aspects of Src kinase activation
Koudelková, Lenka ; Brábek, Jan (advisor) ; Brdička, Tomáš (referee) ; Hejnar, Jiří (referee)
Src kinase plays a crucial role in a multitude of fundamental cellular processes. Src is an essential component of signalling pathways controlling cellular proliferation, motility or differentiation, and is often found deregulated in tumours. Src activity is therefore maintained under stringent and complex regulation mediated by SH3 and SH2 domains and the phosphorylation state of tyrosines 416 and 527. Active Src adopts an open conformation whereas inactive state of the kinase is characterised by a compact structure stabilised by inhibitory intramolecular interactions. We identified phosphorylation of tyrosine 90 within binding surface of SH3 domain as a new regulatory switch controlling Src kinase activation. Using substitutions mimicking phosphorylation state of the residue we demonstrated that tyrosine 90 phosphorylation controls Src catalytic activity, conformation and interactions mediated by the SH3 domain, representing a positive regulatory mechanism leading to elevated activation of mitogenic pathways and increased invasive potential of cells. Based on correlation between compactness of Src structure and its catalytic activity, we constructed a FRET-based sensor of Src conformation enabling to measure the dynamics of Src activation in cells with spatio-temporal resolution. We found that...
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Interplay between mutated huntingtin and precancerogenous status in mammalian cells.
Butalová, Nikola ; Baxa, Monika (advisor) ; Koudelková, Lenka (referee)
Huntington's disease is a serious hereditary disorder that causes mortification of neurons. The disease affects individuals around the age of 40. Its characteristics are involuntary movement of the limbs and a progressive dementia. This disorder is currently without any treatment and always ends with patient dying within a period of 15 years after the first symptoms are discovered. Special relation between Huntington's disease and malign neoplasia was observed at the end of the 20th century. This relation shows lower degree of cancer among the patients with this neurodegenerative disorder compared to the general population. An expanded sequence of a CAG section probably protects these persons against advancement of cancer. Creation of an applicable experimental model with characteristic highly resembling human body was necessary for superior research of this disease. This model can be represented by a transgene mini pig carrying a mutated protein huntingtin - tgHD pig. Another model of a mini-pig showing hereditary occurrence of malign lesions - MeLiM pig was created to research cancer. These two specific lines of mini-pigs were crossbred resulting in not only piglets with melanoma and transgene piglets with mtHTT but also transgene piglets having melanoma at the same time. Furthermore the same...
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Physical education for pre-school children with autism spectrum disorder
Koudelková, Lenka ; Strnad, Pavel (advisor) ; Prokešová, Eva (referee)
3 Abstract Theme of thesis: Physical education for pre-school children with autism spectrum disorder Objectives of thesis: The aimt of my thesis was to prepare a methodical guide for teachers and trainers of free time activities for pre- school children with ASD, then to identify, if the bee can contribute to improve the children self-reliance during practice of their motorical skills, if it can have a positive effect on the behaviour of children during work and whether it can help reducing problematic behaviour. Method: The main metod used in the qualitative part of my research was observation participation method, the other informal interview. The data was evaluated using simple analysis and statistical method of frequencies. Results and conclusions: The objective of my thesis is to prepare a methodical guide for teachers and trainers of free time activities dealing with pre-school children with ASD. I found outied, that participation in a bee can contribute to improve the children's self-reliance during practice of their motorical skills, it also positively affected the behaviour of children during work and it helped to reduce problematic behaviour. Key words: Autistic spectrum disorder, physical education, structured teaching, structured movement activity, pre-school children, free time activity
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Structural and regulatory aspects of Src kinase activation
Koudelková, Lenka
Src kinase plays a crucial role in a multitude of fundamental cellular processes. Src is an essential component of signalling pathways controlling cellular proliferation, motility or differentiation, and is often found deregulated in tumours. Src activity is therefore maintained under stringent and complex regulation mediated by SH3 and SH2 domains and the phosphorylation state of tyrosines 416 and 527. Active Src adopts an open conformation whereas inactive state of the kinase is characterised by a compact structure stabilised by inhibitory intramolecular interactions. We identified phosphorylation of tyrosine 90 within binding surface of SH3 domain as a new regulatory switch controlling Src kinase activation. Using substitutions mimicking phosphorylation state of the residue we demonstrated that tyrosine 90 phosphorylation controls Src catalytic activity, conformation and interactions mediated by the SH3 domain, representing a positive regulatory mechanism leading to elevated activation of mitogenic pathways and increased invasive potential of cells. Based on correlation between compactness of Src structure and its catalytic activity, we constructed a FRET-based sensor of Src conformation enabling to measure the dynamics of Src activation in cells with spatio-temporal resolution. We found that...
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Structural and regulatory aspects of Src kinase activation
Koudelková, Lenka
Src kinase plays a crucial role in a multitude of fundamental cellular processes. Src is an essential component of signalling pathways controlling cellular proliferation, motility or differentiation, and is often found deregulated in tumours. Src activity is therefore maintained under stringent and complex regulation mediated by SH3 and SH2 domains and the phosphorylation state of tyrosines 416 and 527. Active Src adopts an open conformation whereas inactive state of the kinase is characterised by a compact structure stabilised by inhibitory intramolecular interactions. We identified phosphorylation of tyrosine 90 within binding surface of SH3 domain as a new regulatory switch controlling Src kinase activation. Using substitutions mimicking phosphorylation state of the residue we demonstrated that tyrosine 90 phosphorylation controls Src catalytic activity, conformation and interactions mediated by the SH3 domain, representing a positive regulatory mechanism leading to elevated activation of mitogenic pathways and increased invasive potential of cells. Based on correlation between compactness of Src structure and its catalytic activity, we constructed a FRET-based sensor of Src conformation enabling to measure the dynamics of Src activation in cells with spatio-temporal resolution. We found that...
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Structural and regulatory aspects of Src kinase activation
Koudelková, Lenka ; Brábek, Jan (advisor) ; Brdička, Tomáš (referee) ; Hejnar, Jiří (referee)
Src kinase plays a crucial role in a multitude of fundamental cellular processes. Src is an essential component of signalling pathways controlling cellular proliferation, motility or differentiation, and is often found deregulated in tumours. Src activity is therefore maintained under stringent and complex regulation mediated by SH3 and SH2 domains and the phosphorylation state of tyrosines 416 and 527. Active Src adopts an open conformation whereas inactive state of the kinase is characterised by a compact structure stabilised by inhibitory intramolecular interactions. We identified phosphorylation of tyrosine 90 within binding surface of SH3 domain as a new regulatory switch controlling Src kinase activation. Using substitutions mimicking phosphorylation state of the residue we demonstrated that tyrosine 90 phosphorylation controls Src catalytic activity, conformation and interactions mediated by the SH3 domain, representing a positive regulatory mechanism leading to elevated activation of mitogenic pathways and increased invasive potential of cells. Based on correlation between compactness of Src structure and its catalytic activity, we constructed a FRET-based sensor of Src conformation enabling to measure the dynamics of Src activation in cells with spatio-temporal resolution. We found that...
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Advances in chemotherapy and novel antitumor drugs
Kraus, Michal ; Kovář, Marek (advisor) ; Koudelková, Lenka (referee)
Cancer is among the leading causes of death worldwide. While some types of cancer became almost entirely curable, majority of malignant tumors are still potentially deadly diseases due to unsensitivity of tumors to conventional chemotherapy or diversity of cancer cells within the tumor and subsequent development of resistance. The underlying mechanism of action of conventional antitumor drugs is mostly related to cell division. DNA damage, inhibition of DNA synthesis and repair or disrupted formation of mitotic spindle are the most common mechanisms. However, it implies that most of the drugs are cytotoxic for rapidly dividing cells in general which results in variety of undesirable side effects for patients. Search for novel anticancer drugs targeting cancer cells more selectively has been point of interest of researchers for decades. Hundreds of new potential anticancer drugs are being described every year, some posessing so far unrecognized mechanisms of action. Process called drug repurposing examines drugs that have already been approved for clinical use in other than oncology field and results into discovering of interesting "novel" anticancer agents. Another general trend is represented by shift towards development of targeted therapy which is slowly replacing traditional cytotoxic...
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Evolution of regulatory mechanisms of EGF receptor activation
Trávníčková, Květa ; Stříšovský, Kvido (advisor) ; Koudelková, Lenka (referee)
Signalling through EGF receptor is crucial both for ontogenesis and for maintaining homeostasis in adult organisms. It is involved in controlling cellular behaviours such as proliferation, migration or differentiation. This thesis provides an insight into evolution of the regulatory mechanisms of EGF receptor activation by discussing their principles in C. elegans, D. melanogaster and H. sapiens sapiens, on the basis of which conclusions about their evolutionary tendencies are made. Attention is focused on the roles of the rhomboid family of proteins, whose activity is tightly associated with EGF receptor signalling. Dysregulation of the EGF receptor unnegligibly contributes to the development of various diseases, mainly many types of cancer, but also schizophrenia, psoriasis and cardiovascular disorders. Experimental results obtained on this field of research therefore have the potential to be applied in drug design.
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Thyroid tumors and their molecular genetic causes.
Šmídová, Barbora ; Dvořáková, Šárka (advisor) ; Koudelková, Lenka (referee)
The aim of this work is to summarize actual literature overview and to collect the up- to-date knowledge on genetic causes of the development of thyroid carcinomas. Thyroid carcinomas represent the most often endocrine malignancy and its incidence is still growing. This work describes all types of thyroid carcinomas derived from different cell types and are distinguished also according to the degree of differentiation. The main genetic changes in the tumor tissues of the medullary, papillary, follicular and anaplastic thyroid carcinoma are described. Thyroid carcinomas occur mostly in sporadic form, rarely as a familial disease. The causes of familial forms of thyroid carcinomas are also summarized and the main mutations in the germ-line DNA are identified. Key words: carcinoma, genetics, mutation, thyroid
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