|
|
Large Extracellular Vesicles in Cell Culture and Blood: Role in Prion Transmission and Detection by Flow Cytometry
Soukup, Jakub ; Holada, Karel (advisor) ; Šebestová Janoušková, Olga (referee) ; Živný, Jan (referee)
Prions (PrP) are the main cause of neurodegenerative diseases such as Scrapie in sheep, bovine spongiform encephalopathy, chronic wasting disease in deer, and Creutzfeldt-Jakob disease in humans. Although the cellular PrP (PrPC ) is involved in many cellular processes, its precise function still needs to be discovered. The disease is caused by the accumulation of a pathological form of PrP (PrPTSE ), which is caused by direct contact of PrPTSE and PrPC . PrP is anchored in the membrane by GPI and can be transmitted by cell-to-cell contact, tunnelling nanotubes, or extracellular vesicles (EVs). EV factions are divided by different biogenesis into exosomes, microvesicles, and apoptotic bodies. PrPTSE was found in exosomes and microvesicles, but these fractions were never compared to each other. The first aim of the doctoral thesis is a comparison of PrP content, prion-converting activity and infectivity in these fractions on CAD5 and N2a-PK1 cellular models of infection. We isolated a fraction of large EVs (20,000× g) and small EVs (110,000× g) by centrifugation from a conditioned medium. We characterised EVs by cryo-electron microscopy and western blot with Alix, TSG-101, CD63, CD9, and HSP70 markers. The contamination from other cellular compartments was checked by calnexin. EV fractions differed...
|
|
|
Diagnostic potential of extracellular vesicles
Klementová, Helena ; Holada, Karel (advisor) ; Živný, Jan (referee)
Extracellular vesicles (EV) are membrane particles produced by cells. Many published papers describe changes in the level of EV in body fluids under various pathological conditions. In our study, we analyzed the numbers of EV in the peripheral blood of patients with multiple sclerosis (MS) and in the umbilical cord blood of preterm newborns with inflammation using flow cytometry. We extensively optimized the detection method and verified the presence of EV in the samples using electron microscopy. We found a reduced number of T-cell and phosphatidylserine-positive EV in patients with newly diagnosed MS compared to healthy blood donors. In MS patients requiring a change in the treatment, the number of B-cell EV was lower compared to neurological controls. Similarly, the level of endothelial EV was lower in MS patients receiving Natalizumab treatment. In the umbilical cord blood of preterm newborns with inflammation, we observed significantly increased numbers of placental, endothelial and platelet EV compared to preterm newborns without inflammation. Compared to the full-term controls, the changes in EV numbers were not significant. The number of endothelial EV in umbilical cord blood correlated with the concentration of IL-6 in the venous blood of newborns. Likewise, the number of platelet and...
|
|
|
The role of exosomes in chronic myeloid leukemia
Březinová, Lenka ; Krijt, Matyáš (advisor) ; Holada, Karel (referee)
Exosomes are extracellular vesicles of a size range 30-150 nm whose function has been explored in chronic myeloid leukemia (CML) due to their role in proliferation of CML cells, remodelling the bone marrow niche, angiogenesis and resistance to treatment with tyrosin- kinase inhibitors (TKIs). Although BCR-ABL kinase is effectively targeted by TKIs, 20-30 % of patients remain resistant to treatment. Resistance of CML cells to TKIs treatment is supported by exosomes. Exosomes transport proteins, nucleic acids, chemokines and small molecules that stimulate anti-apoptotic or suppress pro-apoptotic processes in leukemic cells. Anti-apoptotic processes are especially enhanced by upregulated protein levels: TGF- β1, USP6 and FGF2 and various types of RNA: miR-365, miR-21, Hsa_circ_0058493 and mRNA for BCR-ABL. In contrast leukemic cells tend to reduce the number of pro-apoptotic molecules, including miR-320, miR-328 and miR-146a-5p. Leukemic cells modify the bone marrow microenvironment through exosomes in the way to support their survival and also in order to adjust expression of adhesion and pro- angiogenic molecules. An important role in those processes play miR-126, miR-210 and miR- 92a. Neither the number of processes affected by CML exosomes nor their potential use in the treatment of CML is...
|
|
|
Elements of the pharmaceutics as the inspiration for teaching chemistry
Zachariášová, Miroslava ; Holada, Karel (advisor) ; Liška, František (referee)
AND KEYWORDS This thesis deals with school chemistry experiments that are inspired by pharmacy. The main goal is to create a chemistry experiment set containing instructions for experiments applicable to teaching chemistry at secondary schools or colleges or during leisure time. These experiments are closely connected to various branches of pharmacy. Pharmaceutical substances, industrial processes and technologies are used in them. The introduction of this thesis concentrates on the teaching of chemistry conceptions within the present educational framework, specifically on the methods and forms of teaching as well as on the specific methods of teaching chemistry i. e. chemistry experiments. References to sources of professional and other literature will be mentioned in the main body of the thesis. The enclosed company contacts point to the companies that have pharmaceutical substance at their disposal for school supply and usage. Finally, safety instructions for dangerous chemical substances in relation to school education will be briefly summarized. Keywords: experiment; chemistry; pharmacy; chemical substances; drug; safety at work; inspiration; motivation; methods and forms of teaching; medicine; pharmaceutical products
|
|
|
The Ecochemistry Games
Čermáková, Jana ; Holada, Karel (advisor) ; Pumpr, Václav (referee)
Diploma work deals with the opportunities of using the games at teaching of ecochemistry at the primary school. The method of game is described generally in the work including the system of curriculum documents. The compiled summary of the education games with the ecochemistry themes is introduced in the experimental part of the work. Effectiveness the games has been verified by practised investigation and its results ratified hypothesis that by means of games we can achieve more affictive and longer results with ecochemistry teaching. This summary can be used by teachers as the inspiration for not only chemistry lessons activities. Key words: environmental education, teaching of chemistry and ecochemistry, didactic games, curriculum documents, cross-sectional themes, key competency
|
|
|
Chemical Reactions and Light Energy
Sečková, Jana ; Holada, Karel (advisor) ; Liška, František (referee)
The thesis "Chemical reactions and light energy" deals with specific approach of teachers to students. Physical and chemical principles of chemical reactions and light energy are explained followed by suggestions of implementing these topics to lessons at basic schools, grammar schools and specialized chemistry high schools. In practical pedagogical part educational implementation of subject matter, image subject dictionary and school project of luminiscence are drawn. Practical chemical part includes 20 effective experiments for teachers, students and students with teacher. Demo experiments are organized in well-arranged file index with illustrations added. Selected experiments were tried out. There is suggestion of programme about luminiscence. Picture part includes photographs taken during laboratory works.
|
|
|
Chemistry of Verne's novels
Deutschová, Olga ; Holada, Karel (advisor) ; Hally, Jan (referee)
V současné době je stále obtížnější upoutat pozornost žáků během výuky a vybavit je do života širokým vzdělanostním základem a žádoucími klíčovými kompetencemi, které jsou dnes stavěny do popředí. Proto bych zde ráda nabídla učitelům chemie možný způsob, jakým lze žáky seznámit s učivem. Snažím se nabídnout zcela konkrétní činnosti a zároveň nechávám učitelům volnou ruku v uplatnění a rozvíjení dále uvedených témat (viz tabulka č. 1). Vycházím z toho, že schopnosti, které si žáci osvojí během vyučování, by měly být založeny na jejich znalostech, zkušenostech a hodnotách. Tato diplomová práce nabízí řadu možností, jak lze uchopit výuku chemie a zábavnou formou žákům předat důležité poznatky z tohoto oboru, motivovat je dalšímu studiu a samostatné činnosti. Četba a téma diplomové práce má význam pro mediální výchovu a možnost rozvíjet u žáků klíčové kompetence v rámci RVP a ŠVP. Právě dnes je aktuální dbát důraz na rozvíjení klíčových kompetencí u žáků ZŠ a SŠ. Tato diplomová práce obsahuje řadu konkrétních činností, kterými se učitel může zaměřit na jednotlivé kompetence. Rozhodně sem patří kompetence učení, řešení problémů, komunikativní, kompetence pracovní a sociální kompetence. Zájemci o níže uvedenou problematiku se naučí řešit samostatně běžné pracovní i mimopracovní problémy, využívat prostředky...
|
|
|
Experiments (with mercury) that you will not do
Záhorská, Denisa ; Holada, Karel (advisor) ; Hally, Jan (referee)
V úvodu práce jsem si vymezila cíl, který spočíval ve vytvoření záznamu ukazujícího fyzikální a chemické vlastnosti rtuti a jejích sloučenin, a to v konkrétních pokusech s kapkou rtuti. S realizací tohoto nelehkého úkolu mi velmi pomohl pan Doc. RNDr. K. Holada,CSc. Podstata mé práce spočívala v pokusech s minimálním množstvím rtuti - tzn. semimikro a mikro provedení. Je paradoxem, že zákon o chemických látkách a chemických přípravcích zakazuje nakládání s jakýmkoliv množstvím rtuti a přitom provádění pokusů v rámci této práce v již shora uvedeném množství, je v případě demonstračních pokusů prováděných učitelem v podstatě neškodné. Powered by TCPDF (www.tcpdf.org)
|
|
|
Games and Toys as Chemistry Teaching Aids
Dubská, Martina ; Holada, Karel (advisor) ; Liška, František (referee)
The thesis is devoted to the study of one of the methods of activation - the game. During the game, the students are involved actively, they are motivated and creative. The main aim is to explore the advantages of the introduction of gaming activities in the teaching of chemistry, as well as the overall topic of gaming activities, the conditions for these activities in chemistry teaching and suggestions for their implementation in education. The part of the thesis is complex of 11 precific games and toys, which are prepared for teaching chemistry at the primary and high schools and universities uncommonly. This file contains the rules, methodological instructions, time allocation, and everything needed for the teaching of games. The practical part will also find a list of many previously published and unpublished games and toys, which can serve as inspiration for teachers of chemistry. Making an inquiry carried out in two elementary schools to confirm the presumption that the study results achieved through gaming activities, are more satisfactory than the results, in which gaming activities were not used.
|
|
|
Apoptosis Signating Pathways and Biological Effects of TGFB
Šimáková, Olga ; Fuchs, Ota (advisor) ; Brdička, Radim (referee) ; Holada, Karel (referee)
Apoptosis is necessary for maintaing the integrity of all alive multicellular organisms and therefore needs to be precisely regulated. Very important regulators of apoptosis are pleiotropic cytokines from TFGβ superfamily (e.g. TGFβ, BMP, aktivins), whose signals are transduced by SMAD proteins. Patients with secondary myelodysplasias and acute myeloid leukemias (MDS/AML) frequently exhibit interstitial deletions of the chromosome-5q resulting in hemizygous loss of the transcription transactivator SMAD5. SMAD5 is a member of the signal transducer family conveying the pleiotropic TGFβ/BMP cytokine signals with roles in development, cell growth control, and tumor progression. Consistent Smad5 gene expression in these cell types and the gradual increase in its mRNA and protein levels in a model of induced erythroid differentiation of murine erythroleukemia (MEL) cells suggest a role of the gene in hematopoiesis. We show that bone morphogenetic protein 4 (BMP4) directs Smad5 activation in human hematopoietic cells, as monitored at the levels of protein phosphorylation, nuclear translocation, and specific transcription response. In vitro induction of normal human CD34+ cells by BMP4 results in significantly increased proliferation of erythroid progenitors (BFU-E) and formation of glycophorin- A+ cells, whereas...
|
guest ::
