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Cytotoxic activity of selected Narcissus cultivars in vitro.
Maršálková, Aneta ; Hradiská Breiterová, Kateřina (advisor) ; Jošt, Petr (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany (16-16230) Author: Aneta Maršálková Supervisor: PharmDr. Kateřina Hradiská Breiterová, Ph.D. Title of thesis: Cytotoxic activity of selected Narcissus cultivars in vitro. Key words: Narcissus, Amaryllidaceae alkaloids, cytotoxicity, HepG2 The main focus of this diploma thesis is the screening of summary alkaloidal extracts of selected cultivars of the genus Narcissus for their cytotoxic activity against HepG2 liver carcinoma cell line. In the second phase, the cytotoxic activity of the majorly represented Amaryllidaceae alkaloids (AmA) was screened, followed by an IC50 value determination. All studied alkaloidal extracts underwent also GC-MS and GC-FID qualitative and quantitative analysis to identify as many alkaloids as possible. A total of 23 alkaloids, namely 11,12-didehydroanhydrolycorine, 1-O-acetyl-10-norpluviine, 9-O-methylpseudolycorine, anhydropseudolycorine, assoanine, dehydroassoanine, dihydrolycorine, epi-norgalanthamine, galanthamine, galanthine, haemanthamine, hippeastrine, incartine, caranine, lycoramine, lycoraminone, lycorine, narwedine, norpluviine, pluviine, pseudolycorine, sanguinine, tazettine and several unspecified alkaloids of the homolycorine structural type were...
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The role of mitochondrial metabolism in initiation and adaptation to hypoxic conditions.
Rohlenová, Terezie ; Novák, Petr (advisor) ; Rohlena, Jakub (referee)
We can meet pathological hypoxia in the cases of hearth attack, ischemic stroke, but also during tumor invasion, thanks to insufficient angiogenesis. The activation of HIF- 1 factor during hypoxic conditions is crucial for the cell survival. This factor modulates energetic metabolism in favor of fast progressing glycolysis (with the contribution of glutaminolysis) which provides to cell enough ATP and "building blocks", while suppressing Krebs cycle and respiration because of shortage of oxygen. The thesis studies energetic metabolism of HepG2 cells (derived from liver carcinoma) which are cultivated in the media with various energetic substrates, i. e. glucose or galactose (always together with glutamine and pyruvate) under the hypoxic conditions (5% O2). HepG2 cells use particularly oxidative metabolism for ATP and "building blocks" production under the normoxic conditions while hypoxic environment causes metabolic shift in glycemic condition. Interestingly, cells cultured in galactose (glutamine) didn't switch the energy metabolism from oxidative to aerobic glycolysis such as cells cultivated in glucose, although HIF-1 factor was stabilized. We found that enhanced activity and integrity of mitochondria, enhanced maximal capacity and reserve capacity of respiration chain correlates with...
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Metabolism of new polysacharidic nanomaterials for biomedicinal applications
Jirátová, Markéta ; Hrubý, Martin (advisor) ; Jirák, Daniel (referee)
Cancer is one of the leading cause of death in modern world, so there is an emerging demand for better diagnostic tools and more specific less toxique therapeutics. Nanoparticles offers characteristics that could fullfill such perspectives. They can easily target tumor by ehanced permeation and retention effect (EPR). Nanoparticles can combine more than one imaging properties, so we can say that they are multimodal, some of them could combine diagnostic and therapeutic molecules in one nanoparticle, which is now highly popular topic of nanoparticles for theranostics . The aim of this thesis was to characterize new multimodal glycogen-based nanoparticle. Glycogen is an ideal structure for nanoparticle design. Glycogen is part of natural dendrimers group which are easily to modify. Glycogen's size is suitable for EPR effect. We have evaluated biological characteristics of five different types of modified glycogen. The in vitro experiments were carried on HepG2 cells. We have set time curve of cellular uptake of this glycogen probes, evaluated cytoplasmatic localization and for the first time we have carried MTT assay. Biodistribution studies on CD1-Nude mice were performed by using non-invasive method for measuring in vivo fluorescence. In conlusion we've provided some of the biological characteristics of new...
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Optimization of culture conditions of human HepG2 liver cells on 4 types of nano- and micro-fiber carriers
Rössner ml., Pavel
The technology describes the optimization of the number of deployed human hepatocytes HepG2, suitable for use in the field of toxicological verification of new drugs or foods, on a 3D culture system consisting of four types of nano- and microfiber carriers. The motivation for finding suitable models for testing potentially genotoxic effects of drugs is the low relevance of tissue models used in the first stages of preclinical evaluation of new substances, as well as the reduction of animal testing.
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Metabolism of new polysacharidic nanomaterials for biomedicinal applications
Jirátová, Markéta ; Hrubý, Martin (advisor) ; Jirák, Daniel (referee)
Cancer is one of the leading cause of death in modern world, so there is an emerging demand for better diagnostic tools and more specific less toxique therapeutics. Nanoparticles offers characteristics that could fullfill such perspectives. They can easily target tumor by ehanced permeation and retention effect (EPR). Nanoparticles can combine more than one imaging properties, so we can say that they are multimodal, some of them could combine diagnostic and therapeutic molecules in one nanoparticle, which is now highly popular topic of nanoparticles for theranostics . The aim of this thesis was to characterize new multimodal glycogen-based nanoparticle. Glycogen is an ideal structure for nanoparticle design. Glycogen is part of natural dendrimers group which are easily to modify. Glycogen's size is suitable for EPR effect. We have evaluated biological characteristics of five different types of modified glycogen. The in vitro experiments were carried on HepG2 cells. We have set time curve of cellular uptake of this glycogen probes, evaluated cytoplasmatic localization and for the first time we have carried MTT assay. Biodistribution studies on CD1-Nude mice were performed by using non-invasive method for measuring in vivo fluorescence. In conlusion we've provided some of the biological characteristics of new...
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The role of mitochondrial metabolism in initiation and adaptation to hypoxic conditions.
Rohlenová, Terezie ; Novák, Petr (advisor) ; Rohlena, Jakub (referee)
We can meet pathological hypoxia in the cases of hearth attack, ischemic stroke, but also during tumor invasion, thanks to insufficient angiogenesis. The activation of HIF- 1 factor during hypoxic conditions is crucial for the cell survival. This factor modulates energetic metabolism in favor of fast progressing glycolysis (with the contribution of glutaminolysis) which provides to cell enough ATP and "building blocks", while suppressing Krebs cycle and respiration because of shortage of oxygen. The thesis studies energetic metabolism of HepG2 cells (derived from liver carcinoma) which are cultivated in the media with various energetic substrates, i. e. glucose or galactose (always together with glutamine and pyruvate) under the hypoxic conditions (5% O2). HepG2 cells use particularly oxidative metabolism for ATP and "building blocks" production under the normoxic conditions while hypoxic environment causes metabolic shift in glycemic condition. Interestingly, cells cultured in galactose (glutamine) didn't switch the energy metabolism from oxidative to aerobic glycolysis such as cells cultivated in glucose, although HIF-1 factor was stabilized. We found that enhanced activity and integrity of mitochondria, enhanced maximal capacity and reserve capacity of respiration chain correlates with...
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