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Differentiation of mesenchymal stem cells
Červenková, Michaela ; Krylov, Vladimír (advisor) ; Čermák, Lukáš (referee)
There are still many diseases which cannot be successfully cured today. For some of them, the answer might be a cell therapy in the form of stem cells applications. Mesenchymal stem cells (MSCs) are multipotent stem cells which can be most frequently isolated from bone marrow and fat tissues. They have several advantages compared to embryonal stem cells, such as a lower risk of tumorigenesis and fewer ethical issues with their acquisition. MSCs also offer the advantage of immunomodulation abilities and autologous transplantation options. MSCs primarily differentiate into mesodermal lines, i.e. into osteocytes, chondrocytes and adipocytes. However, their differentiation into certain cell types of ectoderm and endoderm origin can also be achieved. This bachelor thesis focuses on the characterisation of mesenchymal stem cells and their differentiation into specific cell types. It addresses the factors which induce and influence differentiation process. Identification of desirable combinations of differentiation agents and factors and establishing safe differentiation protocols is necessary for successful utilization of mesenchymal stem cells in applied medicine. Key words: mesenchymal stem cells (MSCs), differentiation, transcription factor, growth factor, multipotency
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Expression of ubiquitin ligases in gastrointestinal tract
Pícková, Markéta ; Sedláček, Radislav (advisor) ; Čermák, Lukáš (referee)
Ubiquitin (Ub) ligases are important regulatory and signalling molecules, which are involved in majority of cellular processes such as differentiation, DNA repair, and regulation of energetic metabolism or immune response. E3 Ubiquitin ligases are also responsible for pathophysiological changes in the organism and their activity is associated with many human diseases including cancers. This makes E3 Ubiquitin ligases to be new diagnostic markers and interesting pharmaceutical targets. Based on previous studies, these enzymes evince very specific expression in the level of tissues or cell populations. Determination of this specific expression is important for a better understanding of their biological function. In this diploma thesis we systematically screened presence of 370 genes of E3-Ub ligases in gastrointestinal tract under physiological conditions and during acute inflammatory damage of distal colon. Obtained data allowed us to select genes, which can play important role in homeostasis as well as pathophysiology and regeneration of gastrointestinal tract. The screening was based on the expression profiling using qPCR, followed by in situ hybridization to determine the exact localization of the gene expression within tissues. From qPCR analysis was predicted hundred thirty seven candidates for...
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Studies towards biological function of ubiquitin E3 ligase Rnf121 in vivo and in vitro
Škarabellová, Kateřina ; Sedláček, Radislav (advisor) ; Čermák, Lukáš (referee)
Although the RING finger protein 121 (RNF121) is a highly conserved E3 ubiquitin ligase from Caenorhabditis elegans to human, its function is poorly understood and in higher eukaryotes it has been studied only at in vitro level. RNF121 has been described to have various functions: i) it was ascribed to function as a broad regulator of NF-κB activation, ii) it was shown to control intracellular trafficking of various membrane proteins, and iii) its downregulation leads to apoptosis. Moreover, RNF121 might have a role in cancer as its expression was found to be 16.4-fold higher in patients suffering from Barrett esophagus (precancerous lesion of esophageal adenocarcinoma) and was even more increased in esophageal adenocarcinoma comparing to healthy population. In addition, RNF121 gene is localized in the candidate region containing breast cancer susceptibility genes. To gain insight into physiological functions of RNF121, Rnf121 knockout mice (Rnf121tm1b(EUCOMM)Hmgu ) were generated in the Czech Centre for Phenogenomics and further studied in our laboratory. Rnf121+ /- intercross breedings showed a prenatal lethal phenotype of Rnf121-/- embryos, which were dying prior embryonic day (E) 11.5. Preliminary experiments carried out in our laboratory showed numerous vascular defects in null mutant embryo,...
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Analysis of software tools to support team cooperation
Čermák, Lukáš ; Šedivá, Zuzana (advisor) ; Pražský, Petr (referee)
This work is focused on the analysis of software tools that can support cooperation within small groups or large companies. The aim of this work is to characterize cooperation activities and describe technology, functions and principles that these tools uses. Another aims of this work are analysis of three software tools for cooperation and creating a decision tool for leaders of team projects and tasks. These objectives were achieved thanks to the knowledge of the studied materials and examination of individual tools and their functions. The benefit of this work is not only a global view of software tools for collaboration, but also creating of inspiration for future analyzes of this kind. Another benefit is creating of materials for business leadership or project manager to select and use appropriate tools and creating of material that initiate to the issues of modern technologies in the field of cooperation.
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The Alto trombone
Čermák, Lukáš ; Pulec, Zdeněk (advisor) ; Junek, Václav (referee) ; Rejlek, Vladimír (referee)
This diploma thesisdeals with the trombone origin and its development from the begenning up to the present time. The trombone, as it is known nowadays, has not actually changed as far as its structure since the 16th century. Except its development this work also tries to comparethe trombone teaching methods (France, Germany, Russia) and some outstanding trombone interpreters. This musica instrument must also be considered as the part and parcel of jezz. The chapter describes the jazz appearance and its styles in Chicago and New Orleans up to its styles in the sixties of the last century.
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Analýza genového clusteru anthramycinových a lincosamidových antibiotik
Jelínková, Markéta ; Koběrská, Markéta ; Čermák, Lukáš ; Kopecký, Jan ; Janata, Jiří ; Spížek, Jaroslav
Lincomycin, produced by Streptomyces lincolnensis, is important, clinically used antibiotic. Its gene cluster consists of twenty-seven putative open reading frames with biosynthetic or regulatory functions and three resistance genes. Comparison of the chemical structure of the lincomycin and celesticetin indicates, that in the biosynthesis of celesticetin the whole propylproline branch is absent and proline is substrate of the penultimate step of biosynthesis, the condensation reaction. Second part of biosynthetic pathway including conversion of L-tyrosine to L-proline should be involved in the biosynthesis of functionally different anthramycin antibiotics. Thus it appears, that genetic information on lincomycin and anthramycin biosynthesis must also share common elements (genes), both biosynthetic and regulatory. The organization of transcription units was found. The analysis of the lincomycin biosynthetic gene transcripts in various cultivation stages revealed the genes with putative regulatory functions which are transcribed sooner. A set of probes based on the sequence of lincomycin biosynthetic cluster was used to search for analogous genes in celesticetin producer Streptomyces caelestis and three producers of anthramycin antibiotics: Streptomyces refuineus, Streptomyces albus and Streptosporangium sibiricum. Hybridization experiments proved presence of several analogues of genes coding for sugar moiety biosynthesis in S. caelestis and analogues of some genes coding for anthramycin antibiotics in different anthramycin producers. A rough restriction map of a part of celesticetin gene cluster presumably responsible for sugar moiety biosynthesis shows the gene arrangement to be very similar to that of lincomycin biosynthetic cluster.
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