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Proteomic analysis in hematology: Identification of alfa2-macroglobulin as a specific carrier for the hormone hepcidin and proteomic analysis of the of leukemic K562 cell differentiation induced by sodium butyrate.
Pešlová, Gabriela ; Vyoral, Daniel (advisor) ; Krijt, Jan (referee) ; Suttnar, Jiří (referee)
The thesis "The proteomic analysis in hematology: Identification of alfa2- macroglobulin as a specific carrier for the hormone hepcidin and proteomic analysis of the leukemic K562 cell differentiation induced by sodium butyrate" describes proteomic approaches, used for the identification and functional characterisation of proteins, which are binding and transporting the iron metabolism regulating hormone hepcidin. Proteomic techniques are also exploited for the identification of proteins, participating in erythroid differentiation of the model cell line K562. In the first section of the thesis, non-denaturing, native techniques, such as chromatography and native electrophoresis are used, in the second section, the control and butyrate - induced K562 cell proteomes are compared using the classical 2D - SDS polyacrylamide gel electrophoresis approach. The methods, described in the thesis are broadening the spectrum of available techniques in experimental hematology. The results, described in this thesis together with the accompanying published manuscripts broaden our knowledge in the function of proteins of iron metabolism and proteins, functioning in erythroid differentiation. Key words: proteomic analysis, hepcidin, alfa2-macroglobulin, iron metabolism, CML, K562, sodium butyrate
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The role of cereblon in lenalidomide therapy of del(5q) myelodysplastic syndrome
Bokorová, Radka ; Fuchs, Ota (advisor) ; Lukačková, Renata (referee) ; Krijt, Jan (referee)
Myelodysplastic syndrome (MDS) with deletion of the long arm of the chromosome 5 (5q - syndrome, del( 5q)) can be characterized by anemia, macrocytosis, a normal or high platelet count, and hypolobulated megakaryocytes in the bone marrow. 5q - syndrome belongs to low - risk MDS, which means low risk to transform to acute myeloid leukemia. 5q - syndrome is ass ociated with female predominance and older age. Another sign is transfusion burden that is treated by erythropoiesis - stimulating agents (ESA) as erythropoietin (EPO). Moreover, the response of MDS patients is around 30 - 60% with the median of the response b eing ~ 24 months. The second line of treatment is lenalidomide (LEN) which is a derivate of teratogenic analog thalidomide. LEN increases erythropoiesis and inhibits the growth of del(5q) erythroid progenitors in vivo and it does not have a significant effe ct on the growth of normal CD34+ progenitors or cytogenetically normal progenitors in MDS with del(5q) clones. LEN is used as therapy in multiple myeloma, myelodysplastic syndrome, and lymphoma. LEN is an expensive agent and not every MDS patient re sponds to this therapy. This is a reason why is a need to find a biomarker for the determination of successful treatment. Some multiple myeloma studies showed that cereblon can be the biomarker...
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Molecular mechanisms of iron transport across plasma membrane in mammalian cells
Balušíková, Kamila ; Kovář, Jan (advisor) ; Ehrmann, Jiří (referee) ; Krijt, Jan (referee)
Iron belongs among the trace elements and its role in humans is irreplaceable. Up to 5 g of iron can be found in adult body distributed among different compounds. Iron ions are therefore essential to all cells of our body and its homeostasis is thoroughly controlled. Iron uptake into the organism is mediated by enterocyte cells in the small intestine, where heme as well as non-heme forms of iron are absorbed. Non-heme iron is absorbed via Dcytb (duodenal cytochrome b), DMT1 (divalent metal transporter 1), ferroportin, hephaestin, and ceruloplasmin molecules. Although these molecules can also participate in non-transferrin- bound iron transport across plasma membranes within the whole organism, mechanisms of this transport are not yet fully elucidated. The aim of the present work was to contribute to our understanding of molecular mechanisms that are involved in non-transferrin-bound iron transport across the plasma membrane of mammalian cells. Our project was focused on the description of non-transferrin- bound iron transport in human cells in vitro and in vivo under conditions of iron deficiency or iron overload. Transformed cell lines, that represent the three main types of cells involved in iron homeostasis, and tissue samples of duodenal biopsies were used as experimental models. The expression...
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Molecular mechanisms of iron transport across plasma membrane in mammalian cells
Balušíková, Kamila ; Kovář, Jan (advisor) ; Ehrmann, Jiří (referee) ; Krijt, Jan (referee)
Iron belongs among the trace elements and its role in humans is irreplaceable. Up to 5 g of iron can be found in adult body distributed among different compounds. Iron ions are therefore essential to all cells of our body and its homeostasis is thoroughly controlled. Iron uptake into the organism is mediated by enterocyte cells in the small intestine, where heme as well as non-heme forms of iron are absorbed. Non-heme iron is absorbed via Dcytb (duodenal cytochrome b), DMT1 (divalent metal transporter 1), ferroportin, hephaestin, and ceruloplasmin molecules. Although these molecules can also participate in non-transferrin- bound iron transport across plasma membranes within the whole organism, mechanisms of this transport are not yet fully elucidated. The aim of the present work was to contribute to our understanding of molecular mechanisms that are involved in non-transferrin-bound iron transport across the plasma membrane of mammalian cells. Our project was focused on the description of non-transferrin- bound iron transport in human cells in vitro and in vivo under conditions of iron deficiency or iron overload. Transformed cell lines, that represent the three main types of cells involved in iron homeostasis, and tissue samples of duodenal biopsies were used as experimental models. The expression...
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Proteomic analysis in hematology: Identification of alfa2-macroglobulin as a specific carrier for the hormone hepcidin and proteomic analysis of the of leukemic K562 cell differentiation induced by sodium butyrate.
Pešlová, Gabriela ; Vyoral, Daniel (advisor) ; Krijt, Jan (referee) ; Suttnar, Jiří (referee)
The thesis "The proteomic analysis in hematology: Identification of alfa2- macroglobulin as a specific carrier for the hormone hepcidin and proteomic analysis of the leukemic K562 cell differentiation induced by sodium butyrate" describes proteomic approaches, used for the identification and functional characterisation of proteins, which are binding and transporting the iron metabolism regulating hormone hepcidin. Proteomic techniques are also exploited for the identification of proteins, participating in erythroid differentiation of the model cell line K562. In the first section of the thesis, non-denaturing, native techniques, such as chromatography and native electrophoresis are used, in the second section, the control and butyrate - induced K562 cell proteomes are compared using the classical 2D - SDS polyacrylamide gel electrophoresis approach. The methods, described in the thesis are broadening the spectrum of available techniques in experimental hematology. The results, described in this thesis together with the accompanying published manuscripts broaden our knowledge in the function of proteins of iron metabolism and proteins, functioning in erythroid differentiation. Key words: proteomic analysis, hepcidin, alfa2-macroglobulin, iron metabolism, CML, K562, sodium butyrate
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