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Methods of preparation and characterization of liposomal systems
Chýlková, Patricie ; Venerová, Tereza (referee) ; Jugl, Adam (advisor)
In this bachelor's thesis the properties of liposomes prepared by reference thin-film rehydration method and Mozafari method were compared. The aim of the experimental part was to determine whether the Mozafari method is convenient alternative for liposome preparation and whether thin-film rehydration and Mozafari methods produce liposomes with the same properties. The prepared liposomes were characterized in terms of size, polydispersity and zeta potential using dynamic light scattering on certain days for one month since the day of preparation. It was determined that electrostatically or sterically stabilized liposomes of identical composition prepared by either method had mutually corresponding properties.
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Membrane modeling in a single-channel microfluidic chip
Tesák, Petr ; Pekař, Miloslav (referee) ; Venerová, Tereza (advisor)
This work deals with the modelation of artificial cellular membranes in a single channel microfluidic chip. It describes the conditions under which the model membranes are created in channels that are of three different widths. A new method of filling such chips was developed where it is possible to check if the chip is ready to use and at the same time fill it up with saline solution. Furthermore data based on the number of created and burst membranes in microfluidic chips was processed. This statistic takes into consideration the state of these artificial membranes in correlation with all of the chosen sizes and flow rates. Altough there were many complications while gathering this data it is still possible to use it to describe all the benefits and disadvantages of these channels.
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Use of advanced fluorescence methods in the study of aggregation
Uhrínová, Alžbeta ; Enev, Vojtěch (referee) ; Venerová, Tereza (advisor)
The diploma thesis deals with the use of advanced fluorescence techniques in the study of aggregation of selected aggregation models (casein and sodium dodecyl sulfate). At the beginning of the research, fluorescent probes Atto 488 and Rhodamine 6G (RH6G) were characterized using a spectrofluorometry, UV-VIS spectrometry, time-resolved fluorescence, and fluorescence correlation spectroscopy (FCS). Aggregation models with the addition of fluorescent probes were characterized using the same advanced fluorescence techniques. The interaction between the fluorescent probe Atto 488 and the aggregation models was not confirmed. The excitation maximum of Atto 488 was at 500 nm and the emission maximum at 520 nm. The lifetime of the fluorescent probe Atto 488 was from 4,20 to 4,82 ns and the hydrodynamic radius was determined to 0,6 nm. Rhodamine 6G had an excitation maximum at 525 nm and an emission maximum at 555 nm. The lifetime of the fluorescent probe RH6G was from 3,99 to 5,27 ns. The intensity of RH6G was found to decrease upon addition of a higher concentration of casein, resulting in quenching. The hydrodynamic radius of the RH6G probe was determined to 0,6 nm. With the addition of casein using the maximum entropy method (MEM) evaluation was from 0,5 to 534 nm. Stable SDS micelles had a hydrodynamic radius by MEM evaluation of 2,4 nm.
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Rheological characterization of polysaccharide gels with incorporated vesicles
Kalendová, Lucie ; Smilek, Jiří (referee) ; Venerová, Tereza (advisor)
This diploma thesis deals with the rheological characterization of polysaccharide gels with incorporated vesicular systems. Positively charged Ion Pair Amphiphile vesicles (IPA) consisting of cetrimethylammonium bromide, sodium dodecyl sulfate, dioctadecyldimethylammonium chloride and cholesterol were chosen for the incorporation. Other vesicles used for incorporation were oppositely charged liposomes made of phosphatidylcholine (DPPC), phosphatidic acid (PA) and cholesterol. The thesis is divided into two sections. The first one deals with preparation, characterization and stability control (temperature and time stability) of both vesicular systems by using dynamic light scattering (DLS). In this part, the vesicles’ stability was confirmed which made them perfect candidates for the following incorporation. The second part of the thesis describes the preparation and rheological characterization of selected gels. Three types of gels were prepared in total, specifically 1% and 2% gellan gum gels and 4% hyaluronic gel. All of them were studied in water, as well as in solutions of IPA vesicles or DPPC liposomes, both of same concentration (1 mmoll-). The subsequent rheological characterization including amplitude, frequency, and stress relaxation sweeps demonstrates there is no significant difference between reference gels and those containing vesicles. This indicates that vesicles used in this thesis were only freely stored in the gel networks, meaning the structure of the gels didn’t affect their transport properties. This is a significant information for using these systems in drug delivery.
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Isolation of photosystem II particles and their characterization
Svitačová, Denisa ; Pekař, Miloslav (referee) ; Venerová, Tereza (advisor)
This master's thesis deals with the isolation and characterization of spinach thylakoid membranes and photosystem II particles. Based on the research, two procedures were chosen for the isolation of spinach thylakoid membranes and photosystem II particles with an emphasis on preserving their native properties. Subsequently, the characterizations were carried out, focusing on the spectroscopic properties of the isolated samples. UV-VIS spectrometry was used to determine the ratio of chlorophyll a to chlorophyll b in samples of thylakoid membranes and photosystem II particles. The success of the isolations was confirmed by fluorescence methods. The excitation and emission spectra of the samples were measured by the steady-state fluorescence spectroscopy, and the fluorescence lifetimes of the samples of thylakoid membranes and photosystem II particles were determined by time-resolved fluorescence. The isolated samples were afterwards visualized with a fluorescence microscope with a CCD camera and the FCS technique.
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Humic substances in cosmetics: characterization and stability studies of lip balm
Dostálová, Tereza ; Venerová, Tereza (referee) ; Enev, Vojtěch (advisor)
This diploma thesis deals with the creation of lip balm containing humate to achieve an antiviral effect against the Herpes simplex virus as one of the other possible treatment options for this disease. To visually enhance the product, a dye was prepared from beetroot, the characterization of which is also part of the work. The dye was separated into its individual components by thin-layer chromatography and analyzed by FTIR to describe the structural molecular units it contains. The lip balm itself was prepared in several variants, containing both humate and dye, containing only humate or dye, and in the form of a "blank" sample. These samples were subjected to a long-term four-month stability study, where two different storage conditions were used. The analysis consisted in the assessment of organoleptic properties, spreading properties, determination of the linear viscoelastic area, flow point, melting point and bioburden. For greater specificity, the effect of dye and humate was studied using diffusion and antimicrobial tests. At the end of the study, a sensory analysis was carried out to determine the possible success of the product among consumers.
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Hydrogels with incorporated vesicular systems
Kalendová, Lucie ; Pekař, Miloslav (referee) ; Venerová, Tereza (advisor)
This bachelor thesis deals with the incorporation of vesicular systems into polysaccharide – based hydrogels. The chosen vesicular systems were Ion Pair Amphiphile vesicles (IPA), which consist of cetrimethylammonium bromide, sodium dodecyl sulfate, dioctadecyldimethylammonium chloride and cholesterol. The work is divided into two parts. In the first part, interactions between IPA vesicles and sodium hyaluronan and interactions between IPA vesicles and sodium polystyrene sulfonate were studied. Even though interactions occured, it was not possible to prepare a hydrogel based on these interactions. The second part of this work deals with incorporation of IPA vesicles into an agarose – based gel when heated. Particles of two different concentrations (1 mmol·l1 and 2 mmol·l1) were used. The effect of the IPA particles on the properties of the gel was observed with rheological measurements. The results showed that the gel with incorporated particles has a longer linear viscoelastic region and that there were no interactions between the IPA particles and the gel. The particles only fill the gaps of the gel network. Different behaviour within the gels with different concentrations of IPA vesicles was not observed.
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Interactions and aggregation in hyaluronan-aminoacid-surfactant systems
Venerová, Tereza ; Velebný, Vladimír (referee) ; Pekař, Miloslav (advisor)
The effect of native hyaluronan addition on CMC of excluded surfactants (Tween 20, BETADET THC 2, SDS, CTAT a CTAB) in physiological solution (0,15 M NaCl) has been investigated by fluoresence spectroscopy with Pyrene and Nile red as probes. The greatest influence on CMC has been observed in systems with cationic surfactant. System of cetyltrimethylamonnium bromid has been closely studied. Solubilization experiments with a hydrophobic dye Sudan red has been realized and aggregation number of this system has been determinated via fluorescence quenching with cetylpyridinium chloride as quencher and Pyrene as fluorescence probe. Addition of hyaluronan (native or hydrophobicaly modified) reduces aggregation number of system.
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Characterization of microfludic chips
Vejrosta, Jakub ; Enev, Vojtěch (referee) ; Venerová, Tereza (advisor)
Nowadays, when we are moving to smaller and smaller scales, microfluidics is being used, which helps us to move from fluid devices to the dimensions of microfluidics, which may have a great application in the future. This work deals with the correct preparation and production of a microfluidic chip. Which consist of two parts. One part is the polydimethylsiloxane (PDMS) part of the chip, and the other is the coverslip. In this work, the production process of this chip is improved. The functionality of these chips and the time dependence of the age of the chip and the polydimethylsiloxane from which the chips are made were tested using fluorescence microscopy.
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Inverse FCS in colloidal systems research
Richterová, Veronika ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on the study of inverse fluorescence correlation spectroscopy, especially with the regard for the usage of different fluorescent probes and different sized analysed particles. At first, the proper concentration of fluorescent probes was determined. In this concentration is the probe considered as a medium surrounding the analysed particles. Based on this concentration, which was determined as 400 M, several sets of samples were prepared. This samples contained different concentration of polystyrene particles of 100 and 500 nm diameter and multilamellar liposomes. Then, the FCS curves of samples with different fluorescent probes were measured. Fluorescein, rhodamine 6G and Atto 488 were used as fluorescent probes. As a result from experiments, it was found, that particles with 100 nm diameter cannot be analysed with none of the fluorescent probes. Inverse FCS method can be applied to systems, that contains particles with 500 nm diameter and fluorescein. Systems with rhodamine 6G have the same behaviour as typical FCS measurement. It is caused by dimerization of this probe and it cannot be used for 500 nm particles. Liposome samples can be established with iFCS method, but the results are biased by random distribution of liposomes size.
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