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Mobilization of electroosmotic flow markers in capillary electrophoresis
Martínková, Eva ; Křížek, Tomáš (advisor) ; Sobotníková, Jana (referee)
Capillary zone electrophoresis is a separation method used to separate charged substances in electric field. Substances migrate to detector due to their own electrophoretic mobility and electoosmotic flow in capillary. Separation is possible because of electroforetic mobility of substances, which is directly proportional to the charge and undirectly proportional to the hydrated radius of ion. This means that neutral molecules form just one neutral zone, which is detected together. Well absorbing neutral substances are thus used as markers of average (mean) EOF. It was found, that some of the markers have dispositions to be mobilized with respect to the EOF. This mobilization is caused by interactions of free electron pair on atoms of nitrogen or oxygen of marker with components of background buffer. The aim of the study was to measure mobilization of some commonly used markers with different cations of acetate buffer and thus find "ideal" marker, whose mobility is almost the same as EOF and find combinations of marker+buffer, which are better to be avoided. Eleven markers in combination with six cations of background electrolyte were studied. We have found, that relative mobilities of some of chosen markers come up to five percent of limit mobilities of small inorganic cations. An appropriate...
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Combination of ellipticine chemotherapy and α5β1 integrin-targeted therapy in human glioblastoma
Martínková, Eva
Charles University Prague Faculty of Science Department of Biochemistry SUMMARY OF THE THESIS COMBINATION OF ELLIPTICINE CHEMOTHERAPY AND α5β1 INTEGRIN-TARGETED THERAPY IN HUMAN GLIOBLASTOMA by MARTÍNKOVÁ Eva Supervisors: Prof. RNDr. Marie STIBOROVÁ, DrSc. Dr. Monique DONTENWILL 2 Table
of
Contents INTRODUCTION............................................................................................................................... 3
AIMS
OF
THE
STUDY ...................................................................................................................... 5
RESULTS............................................................................................................................................. 6
PART
A ................................................................................................................................................................6
PART
B.................................................................................................................................................................6
DISCUSSION....................................................................................................................................... 8
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Study of interaction between protamine and heparin and its applicability in capillary electrophoresis
Martínková, Eva ; Křížek, Tomáš (advisor) ; Kubíčková, Anna (referee)
Heparin is an acid mixture of glycosaminoglycans with high negative charge density which naturally occurs in human body. Due to its ability to bind antithrombin III and thus accelerate inhibition of thrombin it has anticoagulant effect. This is abundantly used in clinical practice for operations, in case of embolia or heart-attacks. Protamine is a mixture of small basic peptides, which is used in clinical practice as a heparin antidote. The interaction between heparin and protamine is electrostatic and is also used for determination of heparin in human plasma or blood using affinity methods. In my study it was found that if protamine and heparin are mixed in one vial, a complex is formed. Its resulting charge depends on concentration ratio of protamine and heparin. On the other hand, in case the protamine is injected as a sample and heparin is added to background electrolyte as a protein-binding ligand, it is possible to determine heparin from decreasing protamine peak area. Because of the complexity of protamine-heparin interaction, tetraarginine was used as structurally close model of protamine to increase repeatability of measurements. The method for determination of heparin was optimalised. It uses 20 mM or 60 mM ortho-phosphoric acid as background electrolyte, 1 mg/mL solution of tetraarginine...
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Mobilization of electroosmotic flow markers in capillary electrophoresis
Martínková, Eva ; Křížek, Tomáš (advisor) ; Sobotníková, Jana (referee)
Capillary zone electrophoresis is a separation method used to separate charged substances in electric field. Substances migrate to detector due to their own electrophoretic mobility and electoosmotic flow in capillary. Separation is possible because of electroforetic mobility of substances, which is directly proportional to the charge and undirectly proportional to the hydrated radius of ion. This means that neutral molecules form just one neutral zone, which is detected together. Well absorbing neutral substances are thus used as markers of average (mean) EOF. It was found, that some of the markers have dispositions to be mobilized with respect to the EOF. This mobilization is caused by interactions of free electron pair on atoms of nitrogen or oxygen of marker with components of background buffer. The aim of the study was to measure mobilization of some commonly used markers with different cations of acetate buffer and thus find "ideal" marker, whose mobility is almost the same as EOF and find combinations of marker+buffer, which are better to be avoided. Eleven markers in combination with six cations of background electrolyte were studied. We have found, that relative mobilities of some of chosen markers come up to five percent of limit mobilities of small inorganic cations. An appropriate...
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Combination of ellipticine chemotherapy and α5β1 integrin-targeted therapy in human glioblastoma
Martínková, Eva
Charles University Prague Faculty of Science Department of Biochemistry SUMMARY OF THE THESIS COMBINATION OF ELLIPTICINE CHEMOTHERAPY AND α5β1 INTEGRIN-TARGETED THERAPY IN HUMAN GLIOBLASTOMA by MARTÍNKOVÁ Eva Supervisors: Prof. RNDr. Marie STIBOROVÁ, DrSc. Dr. Monique DONTENWILL 2 Table
of
Contents INTRODUCTION............................................................................................................................... 3
AIMS
OF
THE
STUDY ...................................................................................................................... 5
RESULTS............................................................................................................................................. 6
PART
A ................................................................................................................................................................6
PART
B.................................................................................................................................................................6
DISCUSSION....................................................................................................................................... 8
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Combination of ellipticine chemotherapy and a5b1 integrin-targeted therapy in human glioblastoma
Martínková, Eva ; Stiborová, Marie (advisor) ; Souček, Pavel (referee) ; Eckschlager, Tomáš (referee) ; Barberi-Heyob, Muriel (referee)
Charles University Prague Faculty of Science Department of Biochemistry SUMMARY OF THE THESIS COMBINATION OF ELLIPTICINE CHEMOTHERAPY AND α5β1 INTEGRIN-TARGETED THERAPY IN HUMAN GLIOBLASTOMA by MARTÍNKOVÁ Eva Supervisors: Prof. RNDr. Marie STIBOROVÁ, DrSc. Dr. Monique DONTENWILL 2 Table
of
Contents INTRODUCTION............................................................................................................................... 3
AIMS
OF
THE
STUDY ...................................................................................................................... 5
RESULTS............................................................................................................................................. 6
PART
A ................................................................................................................................................................6
PART
B.................................................................................................................................................................6
DISCUSSION....................................................................................................................................... 8
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Preparation of analogues of MRI contrast agents and study of their internalization on cell cultures
Krijt, Matyáš ; Poljaková, Jitka (advisor) ; Martínková, Eva (referee)
Magnetic resonance imaging (MRI) is a commonly used non-invasive method to examine internal organs and soft tissues. In order to increase signal intensity and improve specificity contrast agents (CAs) have been developed, which affect T1 or T2 relaxation times. T1-contrast agents are mostly based on Gd(III) complexes. The aim of this thesis was to prepare analogues to commercial Gd(III)-CAs by changing the central atom for europium and use europium fluorescence properties for observing CAs' cell-distribution. However, due to strong cell auto-fluoresce the fluorescence of complexes did not provide convincing images reporting their biochemical fate. The toxicity of prepared complexes was tested on adherent cell cultures and evaluated by Fluorescence-Activated Cell Sorter (FACS). Internalization of the CAs' analogues was confirmed also by elemental analysis (ICP-MS). (In Czech)
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