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Quantification of T1 for Preclinical MRI
Dvořáková, Lenka ; Macíček, Ondřej (referee) ; Jiřík, Radovan (advisor)
T1 mapping of myocardial tissue is important for diagnostics of myocardial fibrosis. Cardiac magnetic resonance imaging of small animals is challenging due to high heart and respiratory rates. Pulse sequences for T1 mapping are proposed in this thesis based on inversion recovery FLASH and Intragate FLASH. The sequence IR FLASH was compared to the reference sequence RARE on a static phantom. Both sequences were applied for measuring the myocardium of a rat. For T1 quantification a software in Matlab was developed. Using this software, T1 maps of rat myocardium were calculated.
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Cobas 8000 System Monitoring and Optimalisation
Dvořáková, Lenka ; Frančík, Vladimír (referee) ; Sekora, Jiří (advisor)
Cobas 8000 system is modular analytic biochemical analyzer. Depending on the composition of modules Cobas 8000 system allows to determine results of clinical chemistry tests (module c 501 or c 702), immunochemical analysis (module e 602) and the concentration of sodium, potassium and chloride ions (ISE module). The system at The University Hospital Brno is composed of one unit of ISE, two modules c 702 and two modules e 602. Bachelor thesis monitors workflow of the Cobas 8000 system and designs optimization that is tested in operation. Software tools developed in this thesis facilitate inspection of the system and analyse the time response. The time response (TAT, Turn Around Time) is time needed to complete a task. There were two program in Visual Basic created. The first program is intended for appraisal of system TAT. The second program is intended for everyday statistics of laboratory TAT. Optimization of the system is performed using program developed in Matlab. This program simulates the workflow of the system Cobas 8000.
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Monitoring of CAR-T lymphocytes in the course of hemato-oncological therapy
Dvořáková, Lenka ; Souček, Ondřej (advisor) ; Řezáčová, Vladimíra (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author Bc. Lenka Dvořáková Supervisor RNDr. Ondřej Souček, Ph.D. Title of Thesis Monitoring of CAR-T lymphocytes in the course of hemato-oncological therapy The diploma thesis deals with the therapy of T cells with a chimeric antigen receptor (CAR) in the treatment of blood malignancies. Specifically, it focuses on monitoring the population of CAR-T cells in patients over time, after the administration of a therapeutic product. Measurements were made from whole blood samples by measurement on a flow cytometer. The theoretical part is devoted to hematological malignancies: their diagnosis and treatment, as well as to CAR-T cells: history, production, clinical use, side effects and determinations. The experimental part is dedicated to processing of the measured results. The goal of the thesis was to prepare a research on the given topic and evaluate the results of monitoring the population of CAR-T cells over time. The performed technique was able to demonstrate CAR-T cells in the blood of all patients, flow cytometry using a test tube from the company Exbio and using the antiFMC63 scFv antibody from the company Acro Biosystems appears to be a suitable method. The rate of expansion of...
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Role of Next Generation Sequencing in Diagnostics and Prognosis of X-linked Lysosomal Storage Disorders
Řeboun, Martin ; Dvořáková, Lenka (advisor) ; Fajkusová, Lenka (referee) ; Krejčí, Jan (referee)
The characterisation of the molecular-genetic etiology of monogenic diseases includes not only the identification of the pathogenic variant, but also the description of its effect on the RNA structure and stability. Additionally, the X-inactivation plays an important role in X-linked diseases. In the presented thesis, we applied the methods of next generation sequencing to study three lysosomal disorders (mukopolysacharidosis type II, MPS II; Danon disease, DD; Fabry disease, FD). Two methodological approaches have been used: 1) panel sequencing with hybridization probes for identification of single nucleotide variants, small deletions/duplications and structure variants (CNVs) 2) amplicon sequencing for analysis of somatic mosaicism and allele specific expression. The panel sequencing enabled us to confirm the molecular-genetic basis of DD in two patients with the identification of two exons duplication and five exons deletion in LAMP2, respectively. The somatic mosaicism was being analyzed by the amplicon sequencing in families with DD, MPS II. We could identify the first case of somatic mosaicism in a patient with DD. The allele specific expression has enlarged the group of methods used in X-inactivation analysis. Its impact has been proved particularly in minimizing misinterpretation of XCI...
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Molecular genetic analysis in Niemann-Pick type C disease
Marešová, Ivona ; Dvořáková, Lenka (advisor) ; Hřebíček, Martin (referee)
Niemann-Pick disease type C (NPC) is a rare, severe disease with autosomal recessive inheritance. Disease is caused by pathogenic mutations located in genes NPC1/NPC2. These genes encode lysosomal non enzymatic NPC1/NPC2 proteins that are part of lipid transport. As a result of malfunction of these proteins intracellular accumulation of lipids occurs, in particular free cholesterol and glycolipids. Causal therapy is currently still unsatisfactory therefore new therapies are evolved. However these therapies depend on whether the patient cells contain at least residual amount of transcript NPC1 gene. In a group of patiens, for which a fibroblast culture was available, I analyzed the effect of pathogenic mutations on the expression level of the transcript. Results showed that for all pathogenic mutations transcript level is low, but detectable. Moreover, I characterized the structure of the NPC1 gene promoter. By sequence analysis I found polymorphisms rs8099071, rs28403610, rs2981422, rs1652354, rs1788774, rs1788772 in promoter. On the basis of the composition of polymorphisms in individual patiens, I estimate six different haplotypes. I performed mutation analysis in DNA of recently diagnosed patient. I found only one pathogenic mutation p.I1061T (c.3182T> C) in the NPC1 gene. Therefore I tested...
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Thermal analysis of onteraction of epidermal lipids III
Dvořáková, Lenka ; Musilová, Marie (advisor) ; Doležal, Pavel (referee)
Thermal analysis of interaction of epidermal lipids III Lenka Dvořáková, diploma paper, June 2008 Abstract The skin is the organ protecting the human body from unwanted influences from the environment. Physical chemical properties, structure and ordering of the epidermal lipids, mainly in the stratum corneum, play a key role in the barrier function of the skin. The major lipids are ceramides, cholesterol and free fatty acids in this layer. We are used the samples mimic such the most frequently referred composition of the epidermal lipids, contained pseudo-ceramid, oleic acid, cholesterol and cholesterol sulfate. Because the samples with oleic acids can at the higher temperatures influence the behavior of the particular constituents of the mixture, we are also performed the evaluation of the contained simpler samples which compound cholesterol and pseudo-ceramid. We are tried hydrated the mixtures at in vitro environment by various routes. And we are observed their characteristics and interactions by means of the differential scanning calorimetry. We are also deal with the evaluation of the particular raw materials and the double-barrelled mixtures. We are verified and in detail elaborated the operating sequences which are not completely unambiguously referred in the literature by means of these studies.
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The Quality of the Dietary Suplements with Content Skullcap
Dvořáková, Lenka ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Name of the work: The quality of the dietary suplements with content skullcap Author: Lenka Dvořáková The work was targered on the determination of main flavonoids from skullcap and most often incident flavonoids in the other herbs, which have been by part of polykomponents dietary supmements analyzed in this work. Three extracts from each preparation were prepared under different conditions. The highest amount of flavonoids was achieved in most of cases by extraction in ultrasonic bath and for using lower temperature. These extracts were prepared from capsulae durae. By contraries by extract prepared from capsulae moles, were measured levels (values) higher by extract prepared by using of water bath. Between companies were large differences in measured values and so informations featured on covers were too differently large.
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Heat shock proteins - - their role in diagnosis and prognosis of pregnancy related complications
Dvořáková, Lenka ; Hromadníková, Ilona (advisor) ; Růžičková, Šárka (referee)
Heat shock proteins increase their gene expression after exposure of cells or organisms to some forms of stress, which may be high temperature, infection, inflammation, hypoxia, lack of nutrients and water. Stressful situations for the body are also pregnancy-related complications associated with placental insufficiency - preeclampsia and IUGR, as well as other pregnancy-related complications such as fetal growth restriction and gestational hypertension. Therefore, I examined whether the occurrence of pregnancy-related complications (preeclampsia, fetal growth retardation, gestational hypertension) affect the gene expression of heat shock proteins. Five hsp systems was detected, namely Hsp27, Hsp60, Hsp70, Hsp90 and HspBP1 in placental tissue samples and whole maternal peripheral blood. Samples came from women with physiological pregnancy and from women with certain pregnancy-related complications (PE, FGR, GH). RNA was isolated from the samples. Detection of hsp expression was performed by using real-time RT-PCR and the comparative Ct method. Changes in gene expression between the test samples and reference sample were examined. To assess the difference between physiological pregnancies and pregnancies with selected pregnancy- related complications, an analysis of variance (ANOVA) was used....
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