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Study of free sphingoid bases in skin barrier
Jarešová, Zuzana ; Kováčik, Andrej (advisor) ; Vraníková, Barbora (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Author: Zuzana Jarešová Supervisor: PharmDr. Andrej Kováčik, Ph.D. Consultant: PharmDr. Lukáš Opálka, Ph.D. Title of diploma thesis: STUDY OF FREE SPHINGOID BASES IN SKIN BARRIER The skin barrier, localized in the stratum corneum (SC), consists of corneocytes and an intercellular matrix formed from three types of lipids - ceramides, free fatty acids, and cholesterol, represented in an equimolar ratio. The overall arrangement of lipids is organized and highly specialized. Ceramides are structurally formed from the fatty acid acyl attached to a sphingoid base. In minor but not insignificant amounts, free sphingoid bases can also be found in the skin barrier. Several studies show that there is an increased concentration of free sphingoid bases in skin barrier disorders, such as atopic dermatitis. Although it is assumed that the presence of free sphingoid bases affects the skin barrier, it is not elucidated the way of their participation till today. The lack of studies or their diverse results leads us to the main goal of this thesis - to clarify how free sphingoid bases influence the skin barrier. In this work, the model membranes were prepared by the isolation of human SC ex vivo. Sphingosine (S),...
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Study of free sphingoid bases in skin barrier
Jarešová, Zuzana ; Kováčik, Andrej (advisor) ; Vraníková, Barbora (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Author: Zuzana Jarešová Supervisor: PharmDr. Andrej Kováčik, Ph.D. Consultant: PharmDr. Lukáš Opálka, Ph.D. Title of diploma thesis: STUDY OF FREE SPHINGOID BASES IN SKIN BARRIER The skin barrier, localized in the stratum corneum (SC), consists of corneocytes and an intercellular matrix formed from three types of lipids - ceramides, free fatty acids, and cholesterol, represented in an equimolar ratio. The overall arrangement of lipids is organized and highly specialized. Ceramides are structurally formed from the fatty acid acyl attached to a sphingoid base. In minor but not insignificant amounts, free sphingoid bases can also be found in the skin barrier. Several studies show that there is an increased concentration of free sphingoid bases in skin barrier disorders, such as atopic dermatitis. Although it is assumed that the presence of free sphingoid bases affects the skin barrier, it is not elucidated the way of their participation till today. The lack of studies or their diverse results leads us to the main goal of this thesis - to clarify how free sphingoid bases influence the skin barrier. In this work, the model membranes were prepared by the isolation of human SC ex vivo. Sphingosine (S),...
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The study of model membrane systems, proteins and protein-membrane interactions using various fluorescence techniques
Štefl, Martin ; Hudeček, Jiří (advisor) ; Teisinger, Jan (referee) ; Plášek, Jaromír (referee)
Membrane rafts (also referred as nanodomains) are membrane structures responsible for many cell processes. Their characterization is challenging because of the transparency, dynamics and small size of those structures. Moreover, high variability of cells makes their study even more complicated. In order to simplify the studies of membrane processes including the formation of those rafts often model membranes like Giant Unilamellar Vesicles (GUVs) and Supported Phospholipid Bilayers (SPBs) are used. In this Thesis new fluorescent tools for studying such membrane processed were developed, tested, or improved. Specifically, the phasor plot an approach applicable to the analysis of the fluorescence lifetime data, was theoretically and experimentally tested and afterwards applied to the characterization of the membrane nanodomains in GUVs. First, we introduced the phasor plots to the excitation state processes like solvent relaxation and Förster resonance energy transfer (FRET) in lipid vesicles. We also employed the phasor plots in protein-ligand interaction, protein folding and denaturation studies. Finally, the phasor plot analysis of FRET data in combination with Fluorescence Correlation Spectroscopy (FCS) was used in characterization of membrane nanodomains in terms of the size, mobility and...
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