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Synthesis and biochemical characterization of hybrid analogues of human insulin and IGF-2
Povalová, Anna ; Stiborová, Marie (advisor) ; Koberová, Monika (referee)
The ever-increasing occurrence of diabetes mellitus brings about the need for development of new therapeutic agents to provide adequate treatment for patients. An important element in this research area is elucidation how insulin works, mainly in connection with insulin-like growth factors (IGF-1 and IGF-2), which show significant structural homology to each other. In addition, their respective receptors - insulin receptor (IR) and receptor for IGF-1 and IGF-2 (IGF-1R) - exhibit very high similarity. As a result, IGF-1 and IGF-2 can bind to IR and insulin can bind to IGF-1R. Of a particular importance is the high affinity binding of IGF-2 to the isoform A of IR. Unlike insulin, which predominantly mediates glucose entry into cells, IGFs induce growth or mitogenic effects. The finding which structural determinants in insulin and IGFs are responsible for the differences in the activation of their cognate receptors could provide an explanation for different functional responses upon binding of these hormones to different target cells. Understanding of this mechanism could also help in the development of functionally selective analogues of these hormones. The aim of this study was the synthesis and characterization of analogues of human insulin extended at the C terminus of the B chain with the amino...
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Comparison of standardized gait evaluation tests for post-stroke patients from physiotherapy point of view
Koberová, Monika ; Tichá, Monika (advisor) ; Drábová, Zuzana (referee)
Title of bachelor thesis: Comparison of standardized gait evaluation tests for post-stroke patients from physiotherapy point of view Abstract: This bachelor thesis deals with a comparison of standardized gait evaluation tests for post-stroke patients. The thesis consists of theoretical and practical part. The theoretical part is focused on stroke, walking in general, characteristics of hemiparetic gait, standardization and standardized tests. The practical part provides the comparison of standardized gait evaluation tests for post-stroke patients from the viewpoint of essential characteristics and qualitative point of view. This part also contains case reports of two patients, in which the gait was evaluated using the selected standardized tests (10 Meter Walk Test, 2 Minute Walk Test, Timed Up and Go, Dynamic Gait Index) and one non-standardized test (Emory Functional Ambulation Profile). These tests were performed at the beginning and at the end of the therapy. Their results were subsequently evaluated and compared. The interpretation of the results in the context of measurement errors, minimal detectable changes and values describing the improvement or deterioration of patient's condition is emphasized in this bachelor thesis. Key words: gait, gait assessment, standardized tests, stroke, measurement...
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Comparison of standardized gait evaluation tests for post-stroke patients from physiotherapy point of view
Koberová, Monika ; Tichá, Monika (advisor) ; Drábová, Zuzana (referee)
Title of bachelor thesis: Comparison of standardized gait evaluation tests for post-stroke patients from physiotherapy point of view Abstract: This bachelor thesis deals with a comparison of standardized gait evaluation tests for post-stroke patients. The thesis consists of theoretical and practical part. The theoretical part is focused on stroke, walking in general, characteristics of hemiparetic gait, standardization and standardized tests. The practical part provides the comparison of standardized gait evaluation tests for post-stroke patients from the viewpoint of essential characteristics and qualitative point of view. This part also contains case reports of two patients, in which the gait was evaluated using the selected standardized tests (10 Meter Walk Test, 2 Minute Walk Test, Timed Up and Go, Dynamic Gait Index) and one non-standardized test (Emory Functional Ambulation Profile). These tests were performed at the beginning and at the end of the therapy. Their results were subsequently evaluated and compared. The interpretation of the results in the context of measurement errors, minimal detectable changes and values describing the improvement or deterioration of patient's condition is emphasized in this bachelor thesis. Key words: gait, gait assessment, standardized tests, stroke, measurement...
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Synthesis and biochemical characterization of hybrid analogues of human insulin and IGF-2
Povalová, Anna ; Stiborová, Marie (advisor) ; Koberová, Monika (referee)
The ever-increasing occurrence of diabetes mellitus brings about the need for development of new therapeutic agents to provide adequate treatment for patients. An important element in this research area is elucidation how insulin works, mainly in connection with insulin-like growth factors (IGF-1 and IGF-2), which show significant structural homology to each other. In addition, their respective receptors - insulin receptor (IR) and receptor for IGF-1 and IGF-2 (IGF-1R) - exhibit very high similarity. As a result, IGF-1 and IGF-2 can bind to IR and insulin can bind to IGF-1R. Of a particular importance is the high affinity binding of IGF-2 to the isoform A of IR. Unlike insulin, which predominantly mediates glucose entry into cells, IGFs induce growth or mitogenic effects. The finding which structural determinants in insulin and IGFs are responsible for the differences in the activation of their cognate receptors could provide an explanation for different functional responses upon binding of these hormones to different target cells. Understanding of this mechanism could also help in the development of functionally selective analogues of these hormones. The aim of this study was the synthesis and characterization of analogues of human insulin extended at the C terminus of the B chain with the amino...
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Study of the composition and organization of cytochrome P450 system by covalent crosslinking
Koberová, Monika ; Hodek, Petr (advisor) ; Hansíková, Hana (referee)
The system of mixed function oxygenase (MFO system) participates in significant roles in the metabolism of endogenous compounds and xenobiotics. This system contains cytochrome P450, NADPH:cytochrome P450 reductase, and also there are assigned NADH: cytochrome b5 reductase and cytochrome b5. It was proved that cytochrome b5 can stimulate or inhibit cytochrome P450 (CYP)-dependent reactions and even change the ratio of resulting metabolites. The mechanism of cytochrome b5 action has not been fully elucidated yet. Elucidation of protein-protein interactions in MFO system and determination of topology of this system could explain the mechanism of cyt b5 action. The covalent cross- linking technique is suitable method for identifying protein-protein interactions within the membrane. Cytochrome b5 contains 3 methionines and in 2 cases the methionines are localized in a short hydrophobic C-terminal membrane anchor. Interactions with cytochrome P450 in the membrane environment can be identified by substitution of two methionine for photoactivatable analogue of methionine (photo-methionine) and subsequent photoactivation. This work is focused on expression and isolation of photo-cytochrome b5 (photo- cyt b5), cytochrome b5 analogue with incorporated photo-methionine. Conditions for photo-methionine...
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Study of modified amino acid incorporation into cytochrome b5
Koberová, Monika ; Hodek, Petr (advisor) ; Pavlásková, Kateřina (referee)
A cytochrome b5 (cyt b5) can influence cytochrome P450 (CYP)-dependent reactions. In consequence of these reactions cytochrome b5 can participate in substance activation (for example drugs, carcinogens) or it can influence proportions of formed metabolites. A mechanism of cyt b5 action has not been fully explained yet. Elucidation of protein-protein interactions in monooxygenase system could explain of the mechanism of cyt b5 action. To study these interactions by using cross-linking techniques is necessary to prepare photolabile cyt b5, which after photoactivation generated higly reactive intermediates which can create a complex with nearby components of the monoogynesase system. This thesis describes how was developed the method for the production of recombinant cyt b5 with modified amino acids. Cyt b5 was expressed in a bacterial strain E. coli BL-21 (DE3) Gold. Before the expression induction, cells were transformed into the limiting medium (DMEM-LM) which did not contain L-leucine and L-methionine. The limiting medium was supplemented by deuterated amino acid d3-methyl-L-methionine and D,L-Leucine. Expressed cyt b5 was isolated and incorporation of d3-methyl-L-methionene has been verified by mass spectrometry. Cyt b5 was obtained mainly as the apoprotein (apo-cyt b5). That is why in this...
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