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Tau protein, a biomarker of Alzheimer's disease: in vitro phosphorylation and tau-reactive antibodies characterization
Hromádková, Lenka ; Bílková, Zuzana (advisor) ; Fialová, Lenka (referee) ; Krejsek, Jan (referee)
Tau protein, a microtubule-associated protein localized in axonal projections of neurons, is a key molecule in the pathology of Alzheimer's disease (AD), the most common cause of dementia in the elderly population. Tau belongs to the group of natively unfolded proteins without globular structure and is prone to numerous posttranslational modifications (PTMs). Under pathological conditions, abnormal PTMs and misfolding of tau protein occurs and leads to oligomerization and aggregation into paired helical filaments forming neurofibrillary tangles, the histopathological hallmark of AD. Currently available drugs applied in AD treatment can only slow the disease progression and those, which halt the AD-specific neurodegenerative processes, are still missing. Very promising and evolving therapeutic approach is immunotherapy, and even immunomodulation by administration of intravenous immunoglobulin (IVIG) products, a reservoir of natural antibodies from the plasma of healthy donors, has been already tested. The discovery of naturally occurring antibodies directed to tau (nTau-Abs) in body fluids of both AD and healthy subjects and their presence in IVIG begin the investigation of their therapeutic potential. Considering a wide range of possible modifications of tau and of various tau species (oligomers,...
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Proteome analysis of secerned proteins of Francisella tularensis
Konečná, Klára ; Hernychová, Lenka (advisor) ; Krejsek, Jan (referee) ; Bílková, Zuzana (referee)
Title of dissertation thesis: Proteome analysis of secerned proteins of Francisella tularensis The facultative intracellular bacterium Francisella tularensis is the causal agent of the infectious disease called tularaemia. Despite the available wide range of new knowledge focused on bacterium Francisella, till now, mechanisms of tularaemia disease pathogenesis were not completely clarified. The contents of our work was based upon analysis and identification of culture filtrate proteins of bacterium F. tularensis of three strains (LVS, FSC00, SchuS4). Among identified proteins, there were seeked protein candidates for secretion and proteins, which could help with explanation of molecular mechanisms of disease pathogenesis caused by F. tularensis. The best protein candidate for secretion is enzyme acid phosphatase with proven important role in bacterium F. tularensis escape from phagosome. The attention was also focused on the new described mechanism of bacterial secretion, mediated by membrane vesicles. By the help of transmission electron microscopy was demonstrated, that F. tularensis of the strains LVS and FSC200 secretes membrane vesicles into extracellular milieu. Key words: Francisella tularensis, secretion systems, cultivation filtrate proteins, secreted proteins, outer membrane vesicles
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Biological activities of Lavandula angustifolia essential oil
Bílková, Zuzana ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Zuzana Bílková, Biological activities of Lavandula angustifolia essential oil, Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, thesis author: PharmDr. Jan Martin, PhD., Hradec Králové, 2013, 72 pages. The thesis called "Biological activities of Lavandula angustifolia essential oil" is interested in biological activities of Lavandula angustifolia essential oil, specifically antifungal, antioxidant, anti-inflammatory, cytotoxicity, nematicidal and repellency activities. The aim of work was to find out, if the oil can be used in these indications and become natural substitution to the chemical compound toxic to the environment. Antifungal activity was tested against the most common human patogenes, low toxicity proves that essential oil can be used on human cells without causing any harm. Antioxidant activity was tested on NO reaction, nematicidal activity was tried on proctection of crop where nematodes cause huge economic loss. Repellency activity can protect both decorative and agricultural plants. Key words: Lavandula angustifolia, biological activities, anti-inflammatory, antifungal, cytotoxicity, repellency, antioxidant, nematicidal, essential oil.
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The experience of parents of children with problem and disordered behavior
Voldřich, Michal ; Viktorová, Ida (advisor) ; Němec, Zbyněk (referee) ; Bílková, Zuzana (referee)
The presented dissertation focuses on the experience of parents of children with problem and disordered behavior. We build on research focused on parenting children with ADHD, with the intention of broadening our view to include the social conflict dimension in children's behavior. In the theoretical part, we examine how the categories of problem and disordered behavior are captured in medical and special pedagogical classifications. Furthermore, we elaborate on research focusing on parental experience. The research itself is based on qualitative methodology, on repeated interviews with the parents of children and their analysis through the methods of grounded theory. We are particularly interested in how the socially conflictual and medicalized dimension of problem behavior enters into experience. KEYWORDS parental experience, problem and disordered behavior, ADHD, qualitative methodology
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Segmentace mikroskopických snímků pomocí level-set metod
Bílková, Zuzana ; Kučera, Václav (advisor)
Název práce: Segmentace mikroskopických snímků pomocí level-set metod Autor: Zuzana Bílková Katedra: Katedra numerické matematiky Vedoucí diplomové práce: RNDr. Václav Kučera, Ph.D., KNM, MFF UK Konzultant: RNDr. Jindřich Soukup, ÚTIA, AV ČR Abstrakt: Tato diplomová práce představuje novou metodu pro segmentaci snímků pořízených mikroskopem s fázovým konrastem. Cílem je oddělit buňky od pozadí. Algoritmus je založen na variační formulaci level set metod, tedy na minimalizaci funkcionálu popisujícího level set funkci. Funkcionál je minimalizován gradientním tokem popsaným evoluční parciální diferenciální rovnicí. Nejdůležitější nové myšlenky jsou inicializace pomocí prahování a nové členy ve funkcionálu, které zrychlují konvergenci a zpřesňují výsledky. Také jsme použili nové funkce napsané v jazyce C k počítání gradientu a Laplaceova operátoru. Tato implementace je třikrát rychlejší než standardní funkce v MATLABu. Dosáhli jsme lepších výsledků než algoritmy, se kterými jsme metodu porovnávali. Klíčová slova: Segmentace, level set metody, aktivní kontury Title: Segmentation of microscopic images using level set methods Author: Zuzana Bílková Department: Department of Numerical Mathematics Supervisor: RNDr....
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Possibilities of flow cytometry in analysis of cellular response to genotoxic stress
Havelek, Radim ; Řezáčová, Martina (advisor) ; Bílková, Zuzana (referee) ; Štětina, Rudolf (referee)
Possibilities of flow cytometry in analysis of cellular response to genotoxic stress Flow cytometry is a modern tool for interrogating the immunophenotype and functional characteristics of cells. It is a technique of quantitative single cell analysis that works by sensing optical properties of cells in a flow stream with laser beam. Analysis and differentiation of the cells is based on size, granularity, and whether the cell is carrying fluorescent molecules in the form of either conjugated antibodies or dyes. The cells may be stained with fluorescent dyes, incubated with fluorogenic substrate or labelled with fluorochrome-linked antibodies specific for molecules either on the surface or in the intracellular components of the cell. Flow cytometers are multiparameter, recording several measurements on each cell. Therefore, it is possible to identify a homogeneous subpopulation within a heterogeneous population of cells. When cells are exposed to genotoxic agent, they respond quantitatively and qualitatively according to the absorbed dose and the cell type. To cope with the resulting damage to cellular DNA, the temporary cell-cycle checkpoints and DNA damage response mechanisms are activated to allow more time for effective repair. However, if these mechanisms fail or the damage is irreparable, then...
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Effect of Cryopreservation on Stem Cells
Pilbauerová, Nela ; Suchánek, Jakub (advisor) ; Dušková, Jana (referee) ; Bílková, Zuzana (referee)
Effect of cryopreservation on stem cells Summary Background: The topic of this study is the cryopreservation of dental pulp stem cells (DPSCs). Cryopreservation is a process of sustaining the viability of cells and tissues by freezing and storing them at sub-zero temperatures where biochemical reactions do not occur. It eliminates the need to preserve stem cells through long-term cultures, allows the storage of stem cells for potential future clinical use or clinical studies. However, it is necessary to fully understand any adverse effects on cryopreserved cells for full applicability in preclinical and clinical practice. The purpose of this study was to determine the effect of cryopreservation on DPSCs stored for 6 and 12 months using an uncontrolled-rate freezing technique. Methods: We successfully isolated ten dental pulp stem cell lineages from donors aged 13 - 18 years to be able to observe the effect of an uncontrolled rate freezing technique on cell size, viability, proliferation activity, relative telomere length, and differentiation potential. We used 10 % dimethyl sulfoxide (DMSO) as a cryoprotective agent (CPA). Results and conclusion: According to the data obtained, the uncontrolled-rate freezing technique is not technically demanding, and it is sufficient for the successful cryopreservation of...
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Analysis of proteins and their changes in biological tissues
Kulhavá, Lucie ; Mikšík, Ivan (advisor) ; Coufal, Pavel (referee) ; Bílková, Zuzana (referee)
(EN) Proteins are the important biologically active substances for the organism, their qualitative and quantitative composition determines thein function in the organism. Protein analysis in biological material is an important part of biological and medical research. This dissertation is focused on the optimalization of sample preparation - the biological materials - namely the pre-analytical phase of sample processing and their preparation for their own analases and also introducing new procedures in theie praparation. The conditions of individual analysis approaches were always optimized by means of one- dimensional and two-dimensional gel electrophoresis and mass spectrometry. The obtained results were statistically evaluated and other possible approaches to futher research were outlined. The main biological material studied was human saliva with regard to oral health. Comparative study of human saliva was performed (human saliva obtained from caries-free and caries-susceptible people). The saliva composition of individual fractions was investigated with regard to oral health. The study was carried out to compare differences in the abundances of proteins in the saliva of caries-free and caries-susceptible persons, based on label-free mass spectrometry quantification, and to perform a gender...
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