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Studies of properties of gene products of the Merkel cell carcinoma polyomavirus: Antibody preparation and expression vector construction.
Sauerová, Pavla ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
Merkel cell polyomavirus (MCPyV) is a recently discovered human virus, having it's genome often integrated in a genome of Merkel carcinoma cells. Although this type of carcinoma is not so usual, it is very aggressive and it's incidence has been rising in last few years. It is not surprising that this virus is nowadays in the centre of scientific interest, as well as other pathogens and mechanisms affecting human life. Because the virus was discovered not so long ago, its research has been at the whole beginning. This diploma thesisaims to contribute to the study of this virus from the molecular-virology point of view. A neutralizing monoclonal antibody, type IgG2a, targeted against the main capsid protein of MCPyV, VP1, and recognizing its conformational epitote was prepared. This antibody was then used for a pilot study of VP1 VLPs MCPyV movement in mammalian cells. Results showed that the studied virus, at least particularly, utilizes caveolin-1-carrying vesicles for its movement in cells (colocalisation of VP1 VLPs and caveolin-1 was observedColocalisation with EEA1 marker of early endosomes, LamP2 marker of endolysosomal compartments or with BiP marker of endoplasmic reticulum was sporadic but significant. These preliminary results suggest that MCPyV might utilise an endocytic pathway leading...
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Interference of selected DNA viruses with apoptotic processes
Sauerová, Pavla ; Forstová, Jitka (advisor) ; Štěpánek, Luděk (referee)
This work is focused on selected DNA viruses and some of their mechanisms used for inhibition or induction of the apoptotic processes. The selected DNA viruses are Hepatitis B virus, polyomaviruses, papillomaviruses and herpesviruses. Viruses developed different strategies for fighting the host defense mechanism during their evolution. One of the host defense mechanisms that reacts against virus infection is apoptosis. In case of viruses we can observe the phenomenon of inhibition or induction of apoptosis (which both depend on the life cycle phase of the virus). The purpose of these "fighting" strategies is to ensure successful replication, virus releasing from the cell and finally to let it spread in an organism or among them. Some "fighting" strategies are similar e.g. targeting and manipulation on p53 oncosupresor level or production of Bcl-2 homologs; other strategies are very specific. Certain viruses have mechanisms which allow them to survive in a host organism for a long time.
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Studium interakce buněk s biomimetickým materiálem a jeho využití v biomedicíně
Sauerová, Pavla ; Hubálek Kalbáčová, Marie (advisor) ; Vandrovcová, Marta (referee) ; Jendelová, Pavla (referee)
Biomaterials are considered as very promising tools for regenerative medicine. They have compensatory or supporting function in organism and they are often developed to support specific conventional medical procedures. So-called biomimetic materials are developed to imitate natural environment of organism and to induce positive innate responses of organism. An essential part of biomaterial development is in vitro biological evaluation, which characterizes (often for the first time) the potential of developed material for its clinical application. This Ph.D. thesis deals with in vitro biological evaluation of three different biomimetic materials. In all three cases, the comprehensive evaluation was an integral part of the material development and optimization processes. Each material was in vitro characterised at the level of cell-material interactions with respect to its intended specific application.. In the first part, cell response to potential drug delivery system based on colloidal complexes of cationic surfactants with hyaluronic acid (HyA) was characterized. HyA protection ability and its limits were described; also the role of fetal bovine serum (FBS) in cell response to the stress stimuli was confirmed. Results considered surfactant-HyA complexes as promising system for drug delivery. In...
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Studium interakce buněk s biomimetickým materiálem a jeho využití v biomedicíně
Sauerová, Pavla ; Hubálek Kalbáčová, Marie (advisor) ; Vandrovcová, Marta (referee) ; Jendelová, Pavla (referee)
Biomaterials are considered as very promising tools for regenerative medicine. They have compensatory or supporting function in organism and they are often developed to support specific conventional medical procedures. So-called biomimetic materials are developed to imitate natural environment of organism and to induce positive innate responses of organism. An essential part of biomaterial development is in vitro biological evaluation, which characterizes (often for the first time) the potential of developed material for its clinical application. This Ph.D. thesis deals with in vitro biological evaluation of three different biomimetic materials. In all three cases, the comprehensive evaluation was an integral part of the material development and optimization processes. Each material was in vitro characterised at the level of cell-material interactions with respect to its intended specific application.. In the first part, cell response to potential drug delivery system based on colloidal complexes of cationic surfactants with hyaluronic acid (HyA) was characterized. HyA protection ability and its limits were described; also the role of fetal bovine serum (FBS) in cell response to the stress stimuli was confirmed. Results considered surfactant-HyA complexes as promising system for drug delivery. In...
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Studies of properties of gene products of the Merkel cell carcinoma polyomavirus: Antibody preparation and expression vector construction.
Sauerová, Pavla ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
Merkel cell polyomavirus (MCPyV) is a recently discovered human virus, having it's genome often integrated in a genome of Merkel carcinoma cells. Although this type of carcinoma is not so usual, it is very aggressive and it's incidence has been rising in last few years. It is not surprising that this virus is nowadays in the centre of scientific interest, as well as other pathogens and mechanisms affecting human life. Because the virus was discovered not so long ago, its research has been at the whole beginning. This diploma thesisaims to contribute to the study of this virus from the molecular-virology point of view. A neutralizing monoclonal antibody, type IgG2a, targeted against the main capsid protein of MCPyV, VP1, and recognizing its conformational epitote was prepared. This antibody was then used for a pilot study of VP1 VLPs MCPyV movement in mammalian cells. Results showed that the studied virus, at least particularly, utilizes caveolin-1-carrying vesicles for its movement in cells (colocalisation of VP1 VLPs and caveolin-1 was observedColocalisation with EEA1 marker of early endosomes, LamP2 marker of endolysosomal compartments or with BiP marker of endoplasmic reticulum was sporadic but significant. These preliminary results suggest that MCPyV might utilise an endocytic pathway leading...
|
|
|
Interference of selected DNA viruses with apoptotic processes
Sauerová, Pavla ; Forstová, Jitka (advisor) ; Štěpánek, Luděk (referee)
This work is focused on selected DNA viruses and some of their mechanisms used for inhibition or induction of the apoptotic processes. The selected DNA viruses are Hepatitis B virus, polyomaviruses, papillomaviruses and herpesviruses. Viruses developed different strategies for fighting the host defense mechanism during their evolution. One of the host defense mechanisms that reacts against virus infection is apoptosis. In case of viruses we can observe the phenomenon of inhibition or induction of apoptosis (which both depend on the life cycle phase of the virus). The purpose of these "fighting" strategies is to ensure successful replication, virus releasing from the cell and finally to let it spread in an organism or among them. Some "fighting" strategies are similar e.g. targeting and manipulation on p53 oncosupresor level or production of Bcl-2 homologs; other strategies are very specific. Certain viruses have mechanisms which allow them to survive in a host organism for a long time.
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