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Intrinsic factors in helper T-cells lineage choice
Andreyeva, Arina ; Neuwirth, Aleš (advisor) ; Chmelař, Jindřich (referee)
The process of clonal expansion of T lymphocytes, or T cells, belongs to the basic characteristics of adaptive immunity. A fundamental role in the immune response is played by the CD4+ T cells which are capable of evolving into the different subtypes (for example Th1 or Tfh) that help other types of cells to effectively eliminate pathogens. Each particular subtype activates different arms of the immune system for the most effective clearance of a particular pathogen. The way how the pathogen will be eliminated depends on the type of infection. This thesis aims to analyze relevant literature and known facts about factors that influence functional T-cell differentiation. This thesis will be mainly focused on the question of how much the T-cell receptor's structure or antigen affinity plays a role in this decision- making process. Another point of interest is the capability of T cells from one clone to produce different T helper cell subtypes, or they are preferentially biased towards a single differentiation pathway. Key words: adaptive immunity, CD4+ T cells, TCR, infections, differentiation
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Effect of oxidative stress on the differentiation of mesenchymal stem cells.
Bura, Radek ; Tlapáková, Tereza (advisor) ; Pacherník, Jiří (referee)
Mesenchymal stem cells are capable of forming different types of tissue such as muscle, bone, fat or cartilage tissue, thanks to the ability to divide and transform into another cell type. Mesenchymal stem cells obtained from various tissues are used for cell therapy and regenerative medicine. Knowledge of the influence of various factors on the differentiation of these multipotent cells is important. Currently, little is known about the effect of reactive oxygen species (ROS) on the differentiation potential of mesenchymal stem cells. The aim of this bachelor's thesis is to summarize the knowledge of the effect of oxidative stress on the differentiation of mesenchymal stem cells and the affected signaling pathways.
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Differentiation of mesenchymal stem cells on fibrin assemblies supported by immobilized growth factors FGF2 and VEGF
Musílková, Jana ; Filová, Elena ; Kaplan, Ondřej ; Bačáková, Lucie
Bioartificial heart valves and vascular grafts prepared from decellularized tissues could be recellularized with bone marrow-derived mesenchymal stem cells (MSCs) that are able to differentiate into both smooth muscle cells and endothelial cells. MSCs differentiation is facilitated by sustained release of growth factors. In our study assemblies based on fibrin, fibrin with heparin, fibrin with adsorbed or covalently-immobilized vascular endothelial growth factor A165 (VEGF) or basic fibroblast growth factor (FGF-2) via binding to heparin attached to fibrin have been prepared and were evaluated for their stimulation of MSCs differentiation. We estimated the mRNA expression of endothelial marker CD31 (PECAM1), smooth muscle marker α-actin (ACTA2), osteoblast markers osteocalcin (BGLAP) and alkaline phosphatase (ALP). The gene expression was estimated using RT-PCR on days 1, 7 and 21 after seeding. The cell morphology and viability was evaluated by LIVE/DEAD staining. VEGF, both adsorbed and covalently bound, increased significantly the expression of smooth muscle marker α-actin. The mRNA expression of ACTA2 on day 7 and 21 raised more than 200 times in comparison to control samples (undifferentiated cells before seeding). The ACTA2 gene expression significantly exceeded the expression of all other evaluated genes at all time intervals. Moreover, on day 21, the late smooth muscle marker desmin (DES) was steeply rising in cells cultivated on assemblies containing heparin and covalently bound VEGF. The expression of osteocalcin was minimal. We conclude that fibrin assembly containing covalently bound VEGF is the most convenient for MSCs differentiation towards smooth muscle cells.
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Elementary mathematics of imaging methods for radiology assistants
JELÍNKOVÁ, Barbora
The topic of the bachelor's thesis responded to the finding that there is currently no suitable educational text containing mathematical foundations for the education of radiological assistants and other related fields. The bachelor thesis contains selected areas of mathematics that are necessary to know to understand the physical foundations of imaging methods. The formulation of these areas is appropriate to the needs of future graduates of these fields of study. The goals of the bachelor thesis were the following: G1 Creation of an educational text and examples containing elementary mathematics that are essential to fully understand physical principles of imaging methods. G2 Compilation of a test that will be used to verify the knowledge of mathematics of radiological assistance field students and its graduates, followed by a statistical expression of knowledge level. Based on the goals, the following hypotheses were made: H1 By application of a curricular process a structure of elementary mathematics of imaging methods for radiological assistants can be described. H2a By comparing mathematical structures with components of a radiological assistant´s profile, individual structural levels of elementary mathematics for radiological assistants can be described. H2b According to needs and abilities of radiological assistants, examples of functions, differentiation, integration, and vectors can be chosen. H3 Knowledge of respondents in radiology mathematics will be distributed close to a normal distribution. With regard to the curricular process, a teaching text was compiled together with example illustrations. This step led to the fulfillment of goal G1. Then, a single-choice test of 20 questions was compiled to determine the level of mathematics knowledge across the field of radiological assistance. This step led to the achievement of goal G2. This test was subsequently extended using an online form among radiological assistants and students in this field. Given the above objectives, hypotheses H1, H2a and H2b could be confirmed. The test results were statistically expressed in the practical part of the bachelor thesis. It was confirmed that the empirical distribution of test responses is close to the normal distribution. Hypothesis H3 was also confirmed by this step. The benefits of the bachelor's thesis can be seen in the practical level (construction of an educational text verified by a test survey) and in the theoretical level (verification of the application of the theory of the curricular process).
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Use of Differentation and Individualization in the teaching of foreign languages.
VOPÁLENSKÁ, Aneta
The diploma thesis deals the theme with the use differentiation and individualization in foreign language teaching. This research is divided into theoretical and partical parts. The theoreticla part focuses on concept of differentiation and individualization, their history and mainly of the personality of Václav Příhoda and his concept of differentiation and individualization in education.It also deals with the concept of differentiation and individualization in present education. The partical part is based on the basis of qualitative research survey in the form of interviews that followed after realized German teaching with use of differentiation and individualization at the selected school.The aim of this thesis is to find answers to questions about how this way of teaching is reflected by the pupils themselves, how the teachers and the assistant who is in the lessons as an independent observer.
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Teaching Foreign Languages in Heterogeneous Classes at High Schools - Working with Talented and Gifted Learners
JANČÍKOVÁ, Karolína
This thesis deals with the teaching of languages in the case of gifted students in heterogenous groups in secondary schools in Czech Republic. The theoretical part of the thesis will first introduce the topic of heterogeneity and its different kinds. It will then move onto defining the key concepts of giftedness and specifically linguistic giftedness. The thesis will tackle the issue of the identification of gifted students and will present their possible characteristics. The second section of the theoretical part revolves around teaching differentiation and its various possibilities, which will be illustrated with concrete examples of approaches relevant to students gifted in languages during language classes in heterogenous groups. A further section also deals with the psychological, social and educational difficulties, which these students may have to face.\\ The practical part of the thesis will aim to verify the findings of the theoretical research and answer the research questions through questionnaires surveying the current situation of gifted students in language classes in Czech secondary schools.\\
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Study of adenosine effects on proliferation of BeWo cell line
Papírník, Josef ; Červený, Lukáš (advisor) ; Vokřál, Ivan (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Josef Papírník Supervisor: doc. PharmDr. Lukáš Červený, Ph.D. Title of diploma thesis: Studium of adenosine on proiferation activity in BeWo cell line The placenta is a rapidly developing organ that provides nutrition, protection and environment for the growing fetus. Fetal development is dependent on the supply of nutrients from the mother's blood either by passive diffusion or mediated by transporters. One of the essential nutrients are nucleosides, which are known to promote DNA synthesis and thus the growth of certain tissues. In order to pass across the plasma membrane they need nukleoside transporters (NTs) because of its hydrophilic feature. Additionally Ado takes part in cell signaling. Its wide- ranging effects, including influencing proliferation, are mediated by its binding to adenosine receptors (ARs). The placenta expresses NTs and ARs, which means that it is equipped to uptake nucleosides from maternal blood and has ability to receive signals from the external environment via the adenosine molecule. However, the importance of nucleosides for placental growth has not been investigated yet. The aim of this thesis is to test the effect of nucleosides on the proliferation of...
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Study of adenosine effects on proliferation of JEG-3 cell line
Nguyen, Ngoc Duong ; Červený, Lukáš (advisor) ; Jirkovský, Eduard (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Nguyen Ngoc Duong Supervisor: doc. PharmDr. Lukáš Červený, Ph.D. Title of diploma thesis: Effect of adenosine on the proliferation of JEG-3 cell line Adenosine is a purinergic signaling molecule that is used in nucleic acid synthesis. Transport of hydrophilic nucleosides through the plasma and/or organelle membranes is provided by equilibrative nucleoside transporters (ENTs), members of the SLC29A transporter family, and concentrative nucleoside transporters (CNTs), members of the SLC28A transporter family. The placenta is a complex and rapidly growing organ. It shows some patterns similar to tumors except for the fact that the placenta's growth is fully controlled. It was found that extracellular nucleosides support the proliferation of cancerous and some non-cancerous cells. The placenta expresses high levels of NTs which indicates the placenta's ability to take up the nucleosides from circulation, however, the contribution of this process to placental growth is known. The diploma thesis aims to test the effect of adenosine and other nucleosides on trophoblast proliferation. We evaluated whether adenosine and other nucleosides increase the proliferation of the choriocarcinoma-derived JEG-3 cell...
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Characterization of the effect of human mutated huntingtin on the neuronal stem cell differentiation.
Budková, Kateřina ; Vodičková, Kateřina (advisor) ; Romanyuk, Natalyia (referee)
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expansion of the CAG codon repeat in the huntingtin gene (HTT). This expansion causes a change in the biochemical properties of the huntingtin protein (HTT), its aggregation and cellular toxicity, which leads to the degeneration of brain neurons, especially in the striatum. Induced pluripotent cells (iPSC) derived directly from HD patient cells can serve as a model system for in vitro modeling of this disease. Because neuronal dysfunctions occur in HD patients years before the first clinical symptoms manifest, this model system may help elucidate the mechanisms that precede the onset of the disease. The aim of this thesis was to differentiate iPSCs (derived from fibroblasts of HD patients and healthy controls) into neural stem cells (NSCs) and subsequently into neuronal cell populations and to monitor molecular changes in their differentiation associated with the effect of mutated HTT. The differentiation process was monitored based on selected markers using immunofluorescence, western blot and qRT-PCR. We were able to generate stable NSC lines derived from 3 control and 3 HD iPSC lines. All 6 NSC lines were able to further differentiate into neural populations. At the transcriptional level, we found a higher...
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Immunomodulatory potential of Sertoli cell progenitors in Xenopus tadpoles during the healing of amputated tail
Mertová, Irem ; Krylov, Vladimír (advisor) ; Procházka, Jan (referee)
A cell culture of common Sertoli and peritubular myoid cells progenitors derived from the testes of male X. tropicalis, called XtiSC, was established in the Laboratory of Developmental Biology. XtiSCs exhibit similar properties as more well-known mesenchymal stem cells that are used in cell therapy for their immunomodulatory and proregenerative properties. Microinjection of XtiSC into the dorsal vein of the tail of the tadpoles has increased macrophage numbers 7 days after tail amputation (dpa), both in controls and in tadpoles after depletion of macrophages by the application of clodrosomes. Macrophage depletion also reduces the migratory ability of XtiSC to the site of tail amputation. Macrophage depletion also led to a reduction in the number of satellite cells 1 dpa. On the seventh day after tail amputation and XtiSC injection, there was a significant increase in their number compared to the control group without XtiSC injection. Using DAF-2DA probe, nitric oxide production was confirmed by injected XtiSC and at the injury site. For future research of the proliferation, differentiation and migration of satellite cells in vivo, a vector expressing EGFP under endogenous Pax7 promoter and Katushka-RFP under gama-crystalline promoter was created and integrated into the X. tropicalis genome using...
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