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Haematopoiesis in Sea lamprey
Kovář, Martin ; Bartůněk, Petr (advisor) ; Živný, Jan (referee)
To find out if the haematopoietic system is common feature of vertebrates, we decided to examine haematopoiesis in a sea lamprey (Petromyzon marinus). All blood cells arises from the haematopoietic stem cells in higher vertebrates. We assume that this is common for the higher vertebrates and a jawless vertebrates, but nobody was interested in the jawless haematopoiesis since 1970. Using a reverse genetic, we identify homologues of important hematopoietic of higher vertebrates in transcriptome of the sea lamprey with emphasis on important receptors or transcription factors, because they can be used as the specific markers of different blood cells and their progenitors. Then we use those sequences for cloning, expression measurements and other work. We picked up sea lamprey as model organism because its unique phylogenetic position, important foe evo-devo studies, but also because lack of elementary knowledge about sea lamprey haematopoiesis. Key words: Petromyzon marinus, haematopoiesis, HSC, evo-devo
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High-throughput screening for the discovery of small molecules modulating cell fate
Ribeiro Pombinho, António José ; Bartůněk, Petr (advisor) ; Bařinka, Cyril (referee) ; Jiráček, Jiří (referee)
The discovery of chemical compounds able to modify the way cells proliferate, differentiate or die can lead not only to the formulation of new drugs for disease treatment or prevention but also to their use as biological probes in the study of the molecular pathways involved in these processes. In order to test thousands of these small molecules in cellular assays, instrument automation and assay miniaturization are necessary. In this thesis, applications of High-Throughput Screening campaigns are described. The Hypoxia and Wnt pathways involved in stem and cancer cell proliferation; the differentiation of hematopoietic, neural and mesenchymal stem cells; and the TRAIL pathway leading to selective cancer cells death were the main subjects chosen. With this approach, it was possible to test the effect of small molecules in eukaryotic cells and in unicellular organisms as exemplified by the search of compounds leading to the death of the protozoan parasite Leishmania. Several chemical compounds were identified as active in modulating cell fate. Of remark were: Monensin that inhibits the Wnt pathway and prevents the growth of tumors in a mouse model of colorectal cancer; Homoharringtonine that, only in combination with TRAIL, induces the death of cancer cells implanted in immunodeficient mice; and...
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Mapping of regulatory elements within 5' region of the Disp3 locus
Oltová, Jana ; Bartůněk, Petr (advisor) ; Krásný, Libor (referee)
Dispatched 3 (Disp3), a thyroid hormone-regulated gene, is studied extensively in our laboratory. Phenotype of cells with overexpressed Disp3 and its expression pattern make it a perfect candidate for a molecular link between thyroid hormone action and cholesterol homeostasis in the brain. Moreover, we hypothesize that it might play a role in certain neurodegenerative disorders and brain tumours. This thesis is aimed at the process of regulation of this gene via thyroid hormone receptor (TR), specifically identification of responsive elements of the thyroid hormone receptor that are necessary for the regulation. Also, we searched for elements recognized by liver X receptor (LXR), as LXR binds to the same arrangement of repeats as TR and there are a number of genes regulated by both of them. We combined in silico analysis of the Disp3 locus with reporter luciferase assays. A cluster of six elements identified around the first exon with two of them being conserved among human and mice draw our attention. In order to analyze this sequence in more detail, reporter vectors of various truncations of 3 kb region around exon 1 were constructed and tested in reporter assays. Reporter assays did not reveal any substantial element activated by TR or LXR; on the other hand, region containing repressor element(s)...
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Panels of steroid receptor reporter cell lines for compound profiling and development of selective ligands for estrogen receptor alpha and beta
Sedlák, David ; Bartůněk, Petr (advisor) ; Anzenbacher, Pavel (referee) ; Kasal, Alexander (referee)
of the Ph.D. thesis Steroid hormone receptors represent a major target in the drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic molecules. The first part of this work describes a preparation of two panels of potent, selective and robust luciferase reporter cell lines on the unified cellular background in U2OS osteosarcoma cell line. This system consists of two panels of stable luciferase reporter cell lines for estrogen receptor α (ERα), estrogen receptor β (ERβ), androgen receptor (AR), glucocorticoid receptor (GR), mineralocorticoid receptor (MR) and progesterone receptor (PR). The first panel of reporter cell lines relies on the expression of the chimeric steroid receptors created by the replacement of the N-terminal part of the steroid receptor molecule by Gal4 DNA binding domain (Gal4 DBD) binding to 9 copies of Gal4 upstream activation sequences (Gal4 UAS) in the promoter of the pGL4 luciferase reporter vector. In the second panel of reporter cell lines the activation of either synthetic promoter containing multiple hormone response elements or viral promoter derived from MMTV LTR is mediated by full- length exogenously expressed steroid receptors. We have extensively validated both panels using 28 well established ligands, carefully...
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Regulation of gene expression by thyroid hormone receptors
Oltová, Jana ; Fafílek, Bohumil (referee) ; Bartůněk, Petr (advisor)
Hormones coordinate various processes in living organisms and their action is mediated either by cell membrane receptors or nuclear receptors. Thyroid hormones (THs) have prominent effects on the growth, development, and many aspects of metabolism, embryogenesis and early life. Their receptors (TRs) belong to the nuclear receptor superfamily, whose members function as ligand-activated transcription factors, and depending on the context, they can act both as activators or as inhibitors of transcription. Thyroid hormone receptors are encoded by two genes, TRα and TRβ, and by alternative splicing multiple isoforms are generated. The major form of TR binds to T3-response element as a heterodimer with retinoid X receptor (RXR). TRs are able to bind T3-response elements (TREs) independently of ligand occupancy. TREs contain two or more subsequent half-site sequences of AGGTCA, which are usually arranged as direct repeats with four nucleotide spacing (DR4). TH-responsive target genes are involved in a wide range of cellular pathways. Example of such gene is Disp3, which might link TH and cholesterol metabolism in certain cell types.
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Complete Care of Pacient with Acute Rupture of Aneurysm of Abdominal Aortis
Swárovská, Jana ; Bartůněk, Petr (advisor) ; Grus, Tomáš (referee)
The author introduces in her final work the summary of complex care of patient with rupture of aneuryzm of abdominal aortae in 6 case studies. In comparison with diffrent cases she introduces in summarry the most important aspects of care, which can significantly influence the next prognosis of patient. Similarly as the scheme of today vision of the world also the care about patient is global, complex and holistic. It is not possible to pass over in silence neither of factors of personality, desease, save system and hospital team. On the base of her long standing experience of nurce in acute attendant care in vascular surgery department, the author draw ours attention on the main important moments of care of patient with life - threatened indisposition and indikates the way to the standardization of procedures, of course by observance of individual and empathic approach of sanitary staff. Within work on project of final work the author except of her professional experience uses also scientific methods, as recognizing of reality and verification of hypothesis. (like secondary data analysis)
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Intensive Care of the Paediatric Invasive Meningococal Infection
Šutoriková, Veronika ; Vobruba, Václav (advisor) ; Bartůněk, Petr (referee)
The thesis deals with the issue of the pediatrie invasive meningococcal infection. Theoretical part is focused on the deseription of meningococcal infection, its incidence, epidemiology, clinical picture, complication and consequences. diagnosis, therapeutic and preventions procedures. Also there is an emphasis on early detection and prompt treatment of sepsis shock. The nursing section of the thesis includes comprehensive care of a seven-month-oldinfant. This is a deseriptive report which deseribes the dynamic course of the disease, aggressive treatment including antibiotics, steroids, respiratory support, circulation support, renal replacement therapy and overall therapy of disseminative intravascular coagulation syndrome. Through that care the child survived without any mutilation. The case report focused on the needs of the child and its mother in nursing care. It elaborates the main nursing diagnoses, their targets, plans and realizations.
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Quality Comparison of the Hygienic Hand Care before and after the Introduction of the Education for Intensive Care Unit Employees of University Hospital Olomouc
Svatošová, Petra ; Bartůněk, Petr (advisor) ; Schejbalová, Miriam (referee)
The thesis deals with the problem of prevention of hospital infections on the intensive care units with regard for sanitary hand care of the employees. The aim of the thesis was to find out the quality of sanitary hand care and whether it is possible to positively influence the attitude of employees to it by education. The theoretical part contains the history and characteristics of hospital infections, describes the recommended procedures of hand care and their significance in prevention of hospital infections and also the list of valid legislative standards. The empirical part of the thesis is oriented on comparison of changes in sanitary hand care before and after the education of intensive care employees in University Hospital Olomouc. The research was made by the method of hidden observation of employees and by the method of questionnaires.These methods were used in the work to confirm or disprove the hypothesis set. The result of the findings of this research investigation will be used in the process of improving the hand care hygiene, among others in the context of standardization, and improving the quality of nursing care.
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