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Heat shock proteins - their role in diagnosis and prognosis of pregnancy-related complications
Dvořáková, Lenka ; Hromadníková, Ilona (advisor) ; Holáň, Vladimír (referee) ; Lipoldová, Marie (referee)
Heat shock proteins provide a universal stress response to cells, for example, exposed to high temperature, heavy metals, extreme pH, infection, inflammation, oxygen radicals, or in case of lack of nutrients and oxygen. We assumed that stress conditions associated with pregnancy-related complications will result in a change in gene expression profile of heat shock proteins we selected for the study. Heat shock protein mRNA levels (Hsp27, Hsp60, Hsp70, Hsp90α and HspBP1) were tested in placental tissue, whole peripheral venous blood and maternal plasma in women with the following pregnancy-related complications - preeclampsia (PE), fetal growth restriction (FGR), gestational hypertension (GH), spontaneous preterm birth (PTB), preterm premature rupture of membranes (PPROM) and normal pregnancies. We also investigated whether the severity of the disease had any impact on hsp gene expression in particular biological samples. In placental tissue, overexpression of Hsp27, Hsp90α and HspBP1 was found in patients with mild preeclampsia (that does not require immediate termination of pregnancy if properly treated), and in women with late onset of preeclampsia with clinical manifestation after 34th week of gestation. Concerning preterm birth, overexpression of Hsp27 and Hsp60 was observed in both groups (PTB...
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Preparation and characterization of cell models of lysosomal hereditary diseases - Mucopolysaccharidoses
Presová, Gabriela ; Dobrovolný, Robert (advisor) ; Dvořáková, Lenka (referee)
Mucopolysaccharidoses are a group of diseases that belong to lysosomal storage disorders. A common sign of these monogenic multisystem diseases is a gene mutation leading to a deficiency of the lysosomal enzyme participating in glycosaminoglycan degradation. It results to their accumulation in the tissues and organs, where they cause a progressive damage. There is no efficient treatment available for most mucopolysaccharidoses. Moreover, the research is complicated because of the low prevalence and type of affected tissues. Animal models of these human diseases are used for an evaluation of newly developed therapeutic approaches. However, they also have many limitations due to the different pathogenesis and catabolic pathways of the accumulated substrates between humans and animals. Therefore, animal models are replaced by human cell models. In this thesis, the development of four mucopolysaccharidoses human cell models is reported (MPS IIID, MPS IVA, MPS IVB, MPS VI). Corresponding genes (GNS, GALNS, GLB1, ARSB) were inactivated using CRISPR/Cas9 technology, where plasmids containing specific inserts are delivered to the target human induced pluripotent stem cells (iPSC), using electroporation. Isolated clones, which represent iPSC disease models, were characterized by Sanger sequencing, enzyme...
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Gene expression of mutant alleles and X inactivation pattern in patients with selected X-linked disorders
Černá, Alena ; Dvořáková, Lenka (advisor) ; Sedláček, Zdeněk (referee)
In comparison to men, the number of X-linked genes is doubled in women as they have two chromosomes X while men are hemizygotes for X-linked genes. This imbalance is compensated by X inactivation (XCI) process, also known as primary X-inactivation, occurring in the early stage of embryogenesis. X inactivation is a random process and females are mosaics of two cell populations. The ratio of expressed alleles in women can be random (50:50) or skewed (≥80:20). The skewed X inactivation may occur due to selection when one of the alleles is preferentially inactivated (secondary X inactivation). In this study XCI status in heterozygous females with various severity of phenotypic symptoms and traits in selected X linked inherited metabolic diseases is analysed, with the focus being Fabry disease - the deficiency of the enzyme alpha-galactosidase A encoded by GLA gene. Moreover, XCI in one family with X linked agammaglobulinemia is examined. Mutant alleles and XCI status based on various loci, different methodical approaches and different tissues is subjected to examination. For the first time, the direct analysis of GLA gene transcript to detect the allele ratio was used alongside with the single-nucleotide polymorphisms in the IDS and LAMP genes for allele-specific expression (ASE) and the AR, RP2 and...
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Retrospective-gating Myocardial T1 Mapping in Rats with Doxorubicin Cardiomyopathy
Dvořáková, Lenka
In vivo T1 mapping of rat myocardium can be challenging given to the high heart and respiratory rate and small size of the heart. We propose a new method for in vivo T1 estimation based on retrospectively gated inversion-recovery fast low-angle shot (IR FLASH) pulse sequence. The method was tested on an animal model of dilated cardiomyopathy. The T1 estimates were mostly in line with the effects of doxorubicin and estradiol described in literature.
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Study of the function of the hybrid proline-rich protein family
Dvořáková, Lenka ; Fischer, Lukáš (advisor) ; Vágner, Martin (referee) ; Havel, Ladislav (referee)
Lenka Dvořáková SUI'^,ÍARYoF TřE PH.D. TFDSIS 4. CONCLUSIONS Ancestral HyPRP evolved probably from a lipid transfer protein (LTP) relative that had acquired a sequence encoding a long proline-rich N-terminal domain. The origin of HyPRPs may be one of the evolutionary innovationsof seedplants. Specific sequence is not probably important for formation of the functional 8 CM domain of HyPRPs with exception of eight conserved cysteine residues.which stabilize the tertiary structureof the domain. HyPRPs with long proline-rich N-terminal domains are relatively well-conserved and probably evolutionary more original. Proteins with aýpical N-terminal domains (very short. glycine-rich) evolved apparently relatively recently and independentlyin differentplant speciespossibly by means of shortening, loss or re€uTangements of the ancestral longer proline-rich domains. Glycine-rich domains could originate from proline-rich ones by way of inversion in the coding sequence.This mechanismcould have greatimpact in evolution of cell wall proteins in general, becausethey are often rich eitherin prolineor glycine. N-terminal domains of angiosperm C-type HyPRPs remained relatively well conserved (long and hydrophobic), while the rest of angiosperm HyPRPs has been undergoing rapid and continuous diversification. However, in the...
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Analysis of Czech P2P lending investors' behaviour drivers
Hudcová, Tereza ; Polák, Petr (advisor) ; Dvořáková Švejdová, Lenka (referee)
This empirical study develops an analysis of peer-to-peer lending market in the Czech Republic by analysing uniquely collected dataset from Czech P2P lending platform Zonky and information obtained directly from its investors. The research question investigates, if there exists correlation between peer-to-peer investors' risk attitude, which is inferred by validated survey method, and their real behaviour on platform. The thesis' results show that investors in online environment behave way riskier than they, from the theoretical point of view, actually should. Results also confirmed that investors' risk attitudes are domain specific. Subsequently, OLS estimation method uncovers that the only factor, that is highly statistically significant, in terms of the impact on lenders' expected return, is the interest rate stated at loan request. This finding further supports lenders' inclination to greater risk, resulting in lack of concern with borrower's characteristics.
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Working class colonies of the town Kladno. Contribution to study of live quality of workers in industrial towns during the time of industrialisation
Dvořáková, Lenka ; Matoušek, Václav (advisor) ; Pauknerová, Karolína (referee)
The topic of my diploma thesis is the workers' housing in Kladno. The aim of the thesis is to present information about the quality of life of the workers' social class in industrial cities in the period of industrialization. The thesis consists of two main parts. In the first part, essential terms important for understanding the topic are defined. It also focuses on the development of mining in the region of Kladno, the overview of mining companies mainly in the second half of the 19th century and the overview of the most important mines. Finally, the development of steel production in Kladno is described. The second part focuses on the research of the workers' quality of life, especially of miners and the development of the life quality in the reference period. Several areas of research have been stated. Workers' jobs, family life, impact of work on health of the workers and their families, the workers' social life and the development of the workers movement, living in workers' housing and demographic development of the population in the region of Kladno. All findings are summed up in the final part. Key words: workers' housing, quality of life, industrial revolution, industrialization
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