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Study of the interaction of lung surfactant with selected polymers
Suchá, Klára ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This thesis deals with the study of interactions between pulmonary surfactant (Curosurf) and selected polymers (N,N,N-trimethylchitosan, polyvinylpyrrolidone, sodium hyaluronate). This work also focuses on the inactivation of lung surfactant using bovine serum albumin, which causes an increase in the surface tension of the surfactant, whereby the inactivated surfactant is unable to fulfill its function in the lungs. The addition of polymers to this mixture has been shown to be an effective means of restoring the surface tension of the surfactant. First of all, this work focuses on finding the optimal concentration ratios of polymer and Curosurf, at which their mutual interaction occurs. The method of dynamic light scattering was used to obtain optimal ratios. In the second part, the work is devoted to the determination of surface tension using a du Noüy ring tensiometer. It was found that the addition of all selected polymers led to a reduction of the surface tension of the inactivated surfactant to values close to native Curosurf. This work provided useful information to understand the mechanism of lung surfactant surface tension recovery.
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Electrospinning of ceramic fibers
Nemčovský, Jakub ; Kaštyl, Jaroslav (referee) ; Částková, Klára (advisor)
This diploma thesis focuses on the fabrication of ceramic fibres by electrospinning. The theoretical part of the thesis summarizes the currently available information regarding ceramic fibres, their properties, applications and fabrication. The theoretical part also describes the process of electrospinning as one of the most frequently used methods of nanofibre fabrication, as well as the parametres influencing this process. The experimental part is aimed at the fabrication of ceramic fibres based on titania, pure non-doped zirconia and yttria-doped zirconia by electrospinning and at the characterization of thus fabricated fibres. Ceramic precursors based on propoxide and polyvinylpyrrolidone were subjected to electrospinning. The experimental part of this diploma thesis also describes the influence of precursor composition, process conditions and calcination temperature on the morphology and phase composition of the fibres. Precursors were characterized by viscosity measurements. Thermogravimetric analysis (TGA), Röntgen analysis (RTG) and scanning electron microscopy (SEM) were used to describe the fibres. By performing electrospinning of precursors based on titanium propoxide and subsequent calcination at 500-1300 °C, TiO2 fibres with thickness of 100-2500 nm were fabricated. The phase composition changed with calcination temperature from 500 °C from anatase phase through rutile blend to pure rutile at 900 °C. By performing electrospinning of precursors based on zirconium propoxide and subsequent calcination at 550-1100 °C, 0 – 8 mol% Y2O3 doped ZrO2 fibres with thickness of 50-1000 nm were fabricated. An analysis of fibres based on non-doped ZrO2, calcined at 550 °C showed a composition of predominantly monoclinic phase. An analysis of 3 or 8 mol% Y2O3 doped ZrO2 fibres calcined at 900 °C showed a composition of predominantly tetragonal phase or purely cubic phase, respectively. With the increasing calcination temperature, the morphology of the fibres changed from porous nanostructure to chain-like non-porous structure consisting of micrometer grains of TiO2 or ZrO2. The ZrO2 fibres calcined at 700 °C remained flexible as well as the spun ones, while their fragility increased with the increase in calcination temperature.
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Preparation of expression system of gamma-lactamase and expression testing
Magyerková, Monika ; Ingr, Marek (advisor) ; Šácha, Pavel (referee)
γ-lactamase is an enzyme clearing five-membered lactam cycles. Polyvinylpyrrolidone (PVP) is one of its potential substrates. Degradation of PVP by γ-lactamase is being studied due to its eventual use in waste-water purifying plants. The aim of the work was to prepare a synthetic gene from the bacterium Comamonas acidovorans and to clone it into the expression vector pET22b. PCA method was used for the synthesis of the γ-lactamase gene. 1725 bp long sequence of the γ-lactamase gene was split into two parts (synthons) which were synthesized individually. After the synthesis restriction cleavage and ligation to the vector pUC19 were performed. Competent cells E. coli, strain DH5α, were transformed by the obtained construct. After the sequence confirmation both synthons were cleaved by restriction endonucleases and connected by single-step ligation to the plasmid pET22b. Expression bacterial cells E. coli, strain BL21(DE3)RIL, were transformed by the recombinant plasmid containing the connected synthons and expression of the recombinant γ-lactamase was tested. Sequence of the clone producing a protein of the expected length was confirmed by sequencing analysis. The prepared plasmid will be used for the expression of recombinant γ- lactamase. (In English)
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Electrospinning of ceramic fibers
Nemčovský, Jakub ; Kaštyl, Jaroslav (referee) ; Částková, Klára (advisor)
This diploma thesis focuses on the fabrication of ceramic fibres by electrospinning. The theoretical part of the thesis summarizes the currently available information regarding ceramic fibres, their properties, applications and fabrication. The theoretical part also describes the process of electrospinning as one of the most frequently used methods of nanofibre fabrication, as well as the parametres influencing this process. The experimental part is aimed at the fabrication of ceramic fibres based on titania, pure non-doped zirconia and yttria-doped zirconia by electrospinning and at the characterization of thus fabricated fibres. Ceramic precursors based on propoxide and polyvinylpyrrolidone were subjected to electrospinning. The experimental part of this diploma thesis also describes the influence of precursor composition, process conditions and calcination temperature on the morphology and phase composition of the fibres. Precursors were characterized by viscosity measurements. Thermogravimetric analysis (TGA), Röntgen analysis (RTG) and scanning electron microscopy (SEM) were used to describe the fibres. By performing electrospinning of precursors based on titanium propoxide and subsequent calcination at 500-1300 °C, TiO2 fibres with thickness of 100-2500 nm were fabricated. The phase composition changed with calcination temperature from 500 °C from anatase phase through rutile blend to pure rutile at 900 °C. By performing electrospinning of precursors based on zirconium propoxide and subsequent calcination at 550-1100 °C, 0 – 8 mol% Y2O3 doped ZrO2 fibres with thickness of 50-1000 nm were fabricated. An analysis of fibres based on non-doped ZrO2, calcined at 550 °C showed a composition of predominantly monoclinic phase. An analysis of 3 or 8 mol% Y2O3 doped ZrO2 fibres calcined at 900 °C showed a composition of predominantly tetragonal phase or purely cubic phase, respectively. With the increasing calcination temperature, the morphology of the fibres changed from porous nanostructure to chain-like non-porous structure consisting of micrometer grains of TiO2 or ZrO2. The ZrO2 fibres calcined at 700 °C remained flexible as well as the spun ones, while their fragility increased with the increase in calcination temperature.
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Preparation of expression system of gamma-lactamase and expression testing
Magyerková, Monika ; Ingr, Marek (advisor) ; Šácha, Pavel (referee)
γ-lactamase is an enzyme clearing five-membered lactam cycles. Polyvinylpyrrolidone (PVP) is one of its potential substrates. Degradation of PVP by γ-lactamase is being studied due to its eventual use in waste-water purifying plants. The aim of the work was to prepare a synthetic gene from the bacterium Comamonas acidovorans and to clone it into the expression vector pET22b. PCA method was used for the synthesis of the γ-lactamase gene. 1725 bp long sequence of the γ-lactamase gene was split into two parts (synthons) which were synthesized individually. After the synthesis restriction cleavage and ligation to the vector pUC19 were performed. Competent cells E. coli, strain DH5α, were transformed by the obtained construct. After the sequence confirmation both synthons were cleaved by restriction endonucleases and connected by single-step ligation to the plasmid pET22b. Expression bacterial cells E. coli, strain BL21(DE3)RIL, were transformed by the recombinant plasmid containing the connected synthons and expression of the recombinant γ-lactamase was tested. Sequence of the clone producing a protein of the expected length was confirmed by sequencing analysis. The prepared plasmid will be used for the expression of recombinant γ- lactamase. (In English)
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