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Cell biology of iron transport in plants.
Batík, Adam ; Žárský, Viktor (advisor) ; Tylová, Edita (referee)
Plants use iron as a cofactor of proteins used in photosynthetic systems, electron transport chain and many more. Iron bioavailability for plants in soil is low because it tends to oxidise and create insoluable compounds.For this reasonplants haveevolvedtwo distinct iron uptake mechanisms.Because of the iron toxicitycaused by production ofreactiveoxygen species via the Fenton reaction and the unspecific transport of metals other than iron, plants have to regulate cellular iron concentrationtightly.Theyhave evolved a complex system of signalling networks that has recently begun to uncover. In additionto the regulation ofiron uptake, the plant cell combats iron toxicity by sequestering iron into storage organelles and by chelating it. Iron is essential for seed sprouting but this work is focused on transport of iron into the plant from the soil,subcellulartransportandlongdistance transport ofironin the vasculature.
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WASH complex and its interactome in human pathology
Pácalt, Ondřej ; Libusová, Lenka (advisor) ; Drobná Krejčí, Eliška (referee)
Efficient transport of cargo to its correct destination is required for the proper functioning of eukaryotic cells. Vesicular trafficking is one of the important means of intracellular transport. Impairment of this process often leads to serious pathologies. Sorting and recycling is the crucial part of vesicular trafficking as it enhances its efficiency. The WASH complex has a key role in the regulation of branched actin patches formation. If this occurs on the membrane of endosomes, then it affects sorting, recycling and cargo trafficking. Mutations in the WASH complex or its interacting partners cause diseases such as hereditary spastic paraplegia, Parkinson disease or light intellectual disability. Despite certain advance in the understanding of above-mentioned pathologies, mechanism of the pathogenesis is still elusive. Research in this field can reveal basic molecular mechanisms responsible for the complexity of cargo sorting, recycling and trafficking and thus provide better opportunities for treatment of affected individuals.
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Dynamics of modified diamond nanocrystals in living cells
Majer, Jan ; Libusová, Lenka (advisor) ; Fišer, Radovan (referee)
Nanodiamonds (NDs) are an interesting platform in biological applications and disease treatment. Because of their photoluminescence properties and modifiable surface, they have been investigated as potential carriers for drugs and nucleic acids as well as fluorescent probes. In order to design NDs meeting specifically desired parameters, which would succeed in clinical trials and in medicinal therapy, understanding the mechanism of uptake and intracellular fate of NDs is crucial. The diploma thesis is focused on mechanistic investigation of ND-based nanoparticles delivering nucleic acids to human cells. First, NDs coated with a novel cationic co-polymer were prepared. NDs were then complexed with siRNA in order to transfect siRNA inside U-2 OS cells. NDs proved to be biocompatible and effective transfection particles as observed by qPCR and colorimetric cytotoxicity and cell viability tests. To examine ND uptake by cells, we inhibited endocytosis by specific inhibitors. Obtained results implicated that ND uptake was clathrin- and caveolin dependent. Nonetheless, more than half of NDs was internalized by cells in a different fashion. Some NDs colocalized with early endosomes, lysosomes and caveolin-derived endosomes after internalization. Other NDs resided either in unknown cell structures or escaped from...
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Dynamics of modified diamond nanocrystals in living cells
Majer, Jan ; Libusová, Lenka (advisor) ; Fišer, Radovan (referee)
Nanodiamonds (NDs) are an interesting platform in biological applications and disease treatment. Because of their photoluminescence properties and modifiable surface, they have been investigated as potential carriers for drugs and nucleic acids as well as fluorescent probes. In order to design NDs meeting specifically desired parameters, which would succeed in clinical trials and in medicinal therapy, understanding the mechanism of uptake and intracellular fate of NDs is crucial. The diploma thesis is focused on mechanistic investigation of ND-based nanoparticles delivering nucleic acids to human cells. First, NDs coated with a novel cationic co-polymer were prepared. NDs were then complexed with siRNA in order to transfect siRNA inside U-2 OS cells. NDs proved to be biocompatible and effective transfection particles as observed by qPCR and colorimetric cytotoxicity and cell viability tests. To examine ND uptake by cells, we inhibited endocytosis by specific inhibitors. Obtained results implicated that ND uptake was clathrin- and caveolin dependent. Nonetheless, more than half of NDs was internalized by cells in a different fashion. Some NDs colocalized with early endosomes, lysosomes and caveolin-derived endosomes after internalization. Other NDs resided either in unknown cell structures or escaped from...
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WASH complex and its interactome in human pathology
Pácalt, Ondřej ; Libusová, Lenka (advisor) ; Drobná Krejčí, Eliška (referee)
Efficient transport of cargo to its correct destination is required for the proper functioning of eukaryotic cells. Vesicular trafficking is one of the important means of intracellular transport. Impairment of this process often leads to serious pathologies. Sorting and recycling is the crucial part of vesicular trafficking as it enhances its efficiency. The WASH complex has a key role in the regulation of branched actin patches formation. If this occurs on the membrane of endosomes, then it affects sorting, recycling and cargo trafficking. Mutations in the WASH complex or its interacting partners cause diseases such as hereditary spastic paraplegia, Parkinson disease or light intellectual disability. Despite certain advance in the understanding of above-mentioned pathologies, mechanism of the pathogenesis is still elusive. Research in this field can reveal basic molecular mechanisms responsible for the complexity of cargo sorting, recycling and trafficking and thus provide better opportunities for treatment of affected individuals.
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