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Interaction of animal cells with organic semiconductors
Schildová, Veronika ; Thalerová, Sandra (referee) ; Víteček,, Jan (advisor)
This bachelor´s thesis was focused on study of biocompatibility of organic semiconductors. In introduction, organic semiconductors and their properties, benefits and features of selected representatives were described. In the following part biocompatibility definition and approaches for its assays including tests of cytotoxicity, material stability were presented. The last part was focused on overview of cell cultures, appropriate cultivation conditions and compounds of media for in vitro testing of biocompatibility. Experimental part showed the result of biocompatibility testing of materials. For this purpose murine 3T3 fibroblasts were used.
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Large Extracellular Vesicles in Cell Culture and Blood: Role in Prion Transmission and Detection by Flow Cytometry
Soukup, Jakub
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...
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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...
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Studium vlivu sarkosinu na kalmodulinem-zprostředkovanou vnitrobuněčnou signalizaci
Gerych, Tomáš
The following diploma thesis titled Effects of sarcosine on calmodulin-dependent intracellular signalization is dedicated to analyzation of current findings in the area of increased sarcosine levels effects to calmodulin and calmodulin dependent kinases mediated intracellular signalization and experimental verification of these findings on malignant and non-malignant cell cultures of prostate origin. Use of sarcosine as a possible marker of prostate cancer is an assumption for elaboration of this diploma thesis. Diagnostics of prostate cancer could be simpler and more effective in case of its confirmation as a usable marker by availability of simple home-testing kits reacting on sarcosine level in urine of tested individual.
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Interaction of animal cells with organic semiconductors
Schildová, Veronika ; Thalerová, Sandra (referee) ; Víteček,, Jan (advisor)
This bachelor´s thesis was focused on study of biocompatibility of organic semiconductors. In introduction, organic semiconductors and their properties, benefits and features of selected representatives were described. In the following part biocompatibility definition and approaches for its assays including tests of cytotoxicity, material stability were presented. The last part was focused on overview of cell cultures, appropriate cultivation conditions and compounds of media for in vitro testing of biocompatibility. Experimental part showed the result of biocompatibility testing of materials. For this purpose murine 3T3 fibroblasts were used.
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Microglia control adenosine A2A-receptor mediated astrogliosis
Svobodová, Magdaléna ; Mladěnka, Přemysl (advisor) ; Červený, Lukáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Magdaléna Svobodová Supervisor: Assoc. Prof. Přemysl Mladěnka, Ph.D. Assoc. Prof. Maria da Glória Correia da Silva Queiroz, Ph.D. Title of diploma thesis: Microglia control adenosine A2A-receptor mediated astrogliosis In the central nervous system, astrocytes and microglia are the main cells coordinating the inflammatory response. During inflammation, dying or temporarily damaged cells release ATP, as a danger-associated signal molecule, that contributes to the induction of astrogliosis and promotes clearance of the debris by immune cells such as microglia. Adenosine that results from ATP metabolism also stimulates astrogliosis. However, the effects of adenosine on astrogliosis may be more complex, since it also modulates microglia phenotype and microglia have been shown to prevent excessive astroglial proliferation mediated by nucleotides. In this context, ATP and adenosine are assumed as relevant signalling molecules in the control of astrogliosis and its modulation by microglia. However, it is still unknown whether and how microglia modulate adenosine-mediated astrogliosis. The present study aims to clarify the impact of microglia in the control of adenosine-induced astrogliosis. Two...
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Microglia control adenosine A2A-receptor mediated astrogliosis
Svobodová, Magdaléna ; Mladěnka, Přemysl (advisor) ; Červený, Lukáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Magdaléna Svobodová Supervisor: Assoc. Prof. Přemysl Mladěnka, Ph.D. Assoc. Prof. Maria da Glória Correia da Silva Queiroz, Ph.D. Title of diploma thesis: Microglia control adenosine A2A-receptor mediated astrogliosis In the central nervous system, astrocytes and microglia are the main cells coordinating the inflammatory response. During inflammation, dying or temporarily damaged cells release ATP, as a danger-associated signal molecule, that contributes to the induction of astrogliosis and promotes clearance of the debris by immune cells such as microglia. Adenosine that results from ATP metabolism also stimulates astrogliosis. However, the effects of adenosine on astrogliosis may be more complex, since it also modulates microglia phenotype and microglia have been shown to prevent excessive astroglial proliferation mediated by nucleotides. In this context, ATP and adenosine are assumed as relevant signalling molecules in the control of astrogliosis and its modulation by microglia. However, it is still unknown whether and how microglia modulate adenosine-mediated astrogliosis. The present study aims to clarify the impact of microglia in the control of adenosine-induced astrogliosis. Two...
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Analysis of dynamical interactions of axon shafts and their biopysical modelling.
Šmít, Daniel ; Zápotocký, Martin (advisor) ; Reingruber, Jūrgen (referee) ; Maršálek, Petr (referee)
in English While axon fasciculation plays a key role in the development of neural networks, very lit- tle is known about its dynamics and the underlying biophysical mechanisms. In a model system composed of neurons grown ex vivo from explants of embryonic mouse olfactory epithelia, we observed that axons dynamically interact with each other through their shafts, leading to zippering and unzippering behaviour that regulates their fasciculation. Taking advantage of this new preparation suitable for studying such interactions, we carried out a detailed biophysical analysis of zippering, occurring either spontaneously or induced by micromanipulations and pharmacological treatments. We show that zippering arises from the competition of axon-axon adhesion and me- chanical tension in the axons. This is upheld on quantitative level by conforming change of network global structure in response to various pharmacological treatments, without active involvement of growth cones. The calibrated manipulations of interacting shafts provide qualitative support for the hypothesis, and also allow us to quantify the mechan- ical tension of axons in our system. Furthermore, we introduce a biophysical model of the zippering dynamics, which efficiently serves the purpose of estimating the magnitude of remaining involved...
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Response of animal cells to the extract, obtained from the colony matrix of Pectinatella magnifica
ŠUSTEROVÁ, Karolína
The aim of this thesis was to determine whether colonies freshwater bryozoans Pectinatella magnifica produce biologically active substances. On the basis of knowledge of the course of the growth curves of cell culture mouse fibroblasts were developed models for biological assays to determine the biological potential of substances contained in the extract of lyophilisate colonies of P. magnifica. Tests demonstrated the presence of substances with biological activity. Next part of this work was also survey the occurrence and distribution of the reference type to Vltavotýnsku, including the monitoring of basic parameters of the aquatic environment. It was explored in two years (2012 and 2013) repeated a total of 10 sites on rivers Vltava and Lužnice, the incidence was detected at 5 locations .
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