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Use of nanofluidic mixing for preparation of liposome carriers stained by gadolinium for contrast imaging by magnetic resonance (MRI)
Velínská, Kamila ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis focuses on the preparation of the liposomes, containing lipids with gadolinium, which are used for a contrast magnetic resonance imaging. The liposomes were prepared by the lipid film hydration followed by an extrusion and also by a new nanofluid mixing method on the NanoAssemblr Benchtop. The preparation technology has been optimized for parameters such as the composition of lipids, the flow rate ratio and total flow rate. The method of modification of the liposomes surface by gadolinium complexes has been developed. This method is using a conjugation reaction between the lipids containing cyanuric acid and Gd-DOTA. Prepared Gd-liposomes, which contain gadolinium, were complexly defined by the characterization techniques of DLS and NTA. The morphology of liposomes was observed by TEM and cryo-TEM. Methods for the determination of phospholipid content (Stewart test) and residual water in the lyophylisates of liposomes (Karl-Fischer titration) were used. Gadolinium in liposomal preparations was determined by ICP-OES. Using MR, the concept of gadolinium liposomes was verified and designed for MRI imaging of thrombi. The concept describing the mechanism of liposomes formation based on the experimentally proven existence of a phospholipid bilayer fragment has been developed. This concept is based on the experimentally proven existence of a phospholipid bilayer fragment.
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Use of nanofluidic mixing for preparation of liposome carriers stained by gadolinium for contrast imaging by magnetic resonance (MRI)
Velínská, Kamila ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis focuses on the preparation of the liposomes, containing lipids with gadolinium, which are used for a contrast magnetic resonance imaging. The liposomes were prepared by the lipid film hydration followed by an extrusion and also by a new nanofluid mixing method on the NanoAssemblr Benchtop. The preparation technology has been optimized for parameters such as the composition of lipids, the flow rate ratio and total flow rate. The method of modification of the liposomes surface by gadolinium complexes has been developed. This method is using a conjugation reaction between the lipids containing cyanuric acid and Gd-DOTA. Prepared Gd-liposomes, which contain gadolinium, were complexly defined by the characterization techniques of DLS and NTA. The morphology of liposomes was observed by TEM and cryo-TEM. Methods for the determination of phospholipid content (Stewart test) and residual water in the lyophylisates of liposomes (Karl-Fischer titration) were used. Gadolinium in liposomal preparations was determined by ICP-OES. Using MR, the concept of gadolinium liposomes was verified and designed for MRI imaging of thrombi. The concept describing the mechanism of liposomes formation based on the experimentally proven existence of a phospholipid bilayer fragment has been developed. This concept is based on the experimentally proven existence of a phospholipid bilayer fragment.
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Preparation of fluorescently labeled substrates and determination of transglycosylation activity in plants
Velínská, Kamila ; Ing.Kristína Kováčová (referee) ; Firáková, Zuzana (advisor)
The bachelor's thesis was focused on preparation of fluorescently labeled oligosaccharides and detection of transglycosylating/heterotransglycosylating activities of enzymes from dicotyledons. As the model organisms nasturtium and parsley were used. The quality and composition of labeled acceptor substrates was monitored by TLC and MALDI-TOF MS. The presence of enzymes in the plant seeds was analysed by SDS electrophoresis and IEF-PAGE. The paper method and HPLC were used for the detection of transglycosylating activities. The obtained results suggest that enzymes catalyzing heterotransglycosylating reactions are in the seeds of studied plants.
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