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The effect of Ca2+cations and cholesteryl sulfate on the human skin barrier model
Kelemanová, Aneta ; Pullmannová, Petra (advisor) ; Paraskevopoulou, Anna (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Author: Aneta Kelemanová Supervisor: Mgr. Petra Pullmannová, Ph.D. Title of diploma thesis: The effect of Ca2+ cations and cholesteryl sulfate on the human skin barrier model The structure and composition of the outermost layer of the epidermis, the stratum corneum, form a skin barrier to transepidermal water loss. The main components of the stratum corneum are corneocytes and the intercellular lipid matrix. The intercellular lipid matrix is a mixture of ceramides, cholesterol, and free fatty acids. Cholesterol esters, such as cholesteryl sulfate, are also present. This unique lipid mixture forms a multilamellar layer with short and long crystalline phases with periodicities of approximately 6 and 13 nm. A separated cholesterol phase with a periodicity of about 3,4 nm can also be found in the stratum corneum. The most common lateral packing of the lipid mixture is orthorhombic. Hexagonal or fluid packing is less common. The aim of this diploma thesis was to optimize the conditions for spontaneous arrangement of the isolated human skin lipids in an aqueous medium with the addition of Ca2+ cations and cholesteryl sulfate. A human skin barrier model was created from the prepared lipid dispersion...
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The effect of Ca2+cations and cholesteryl sulfate on the human skin barrier model
Kelemanová, Aneta ; Pullmannová, Petra (advisor) ; Paraskevopoulou, Anna (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Author: Aneta Kelemanová Supervisor: Mgr. Petra Pullmannová, Ph.D. Title of diploma thesis: The effect of Ca2+ cations and cholesteryl sulfate on the human skin barrier model The structure and composition of the outermost layer of the epidermis, the stratum corneum, form a skin barrier to transepidermal water loss. The main components of the stratum corneum are corneocytes and the intercellular lipid matrix. The intercellular lipid matrix is a mixture of ceramides, cholesterol, and free fatty acids. Cholesterol esters, such as cholesteryl sulfate, are also present. This unique lipid mixture forms a multilamellar layer with short and long crystalline phases with periodicities of approximately 6 and 13 nm. A separated cholesterol phase with a periodicity of about 3,4 nm can also be found in the stratum corneum. The most common lateral packing of the lipid mixture is orthorhombic. Hexagonal or fluid packing is less common. The aim of this diploma thesis was to optimize the conditions for spontaneous arrangement of the isolated human skin lipids in an aqueous medium with the addition of Ca2+ cations and cholesteryl sulfate. A human skin barrier model was created from the prepared lipid dispersion...
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New regulatory mechanisms of microtubule nucleation
Černohorská, Markéta ; Dráber, Pavel (advisor) ; Binarová, Pavla (referee) ; Hašek, Jiří (referee)
MT nucleation from γ-tubulin complexes, located at centrosome, is an essential step in the formation of MT cytoskeleton. In mammalian cells, -tubulin is encoded by two genes. We functionally characterized two γ-tubulin proteins and have found that both are functionally equivalent. γ-Tubulin 2 is able to substitute for γ-tubulin 1 in MT nucleation. However, we revealed that unlike TUBG1, TUBG2 expression is downregulated in mouse preimplantation development. Mast cells represent effectors of the allergy reaction. Their activation by antigen induces number of cellular processes such as degranulation, proliferation and cytoskeleton rearrangements. The regulatory mechanisms of MT reorganization during mast cell activation are unknown. We identified new signaling proteins, GIT1 and PIX that interact with - tubulin. Depletion of GIT1 or PIX leads to changes in MT nucleation. GIT1 is phosphorylated on tyrosine and associates with γ-tubulin in a Ca2+ -dependent manner. Our data suggested a novel signaling pathway for MT rearrangement in mast cells where tyrosine kinase-activated GIT1 and βPIX work in concert with Ca2+ signaling to regulate MT nucleation. We tested the capability of GIT1 and PIX to influence -tubulin function in more cell types. We found out that GIT1/βPIX signaling proteins together...
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