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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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Migration of pharmaceuticals in soil systems
Janebová, Denisa ; Pekař, Miloslav (referee) ; Klučáková, Martina (advisor)
The increasing use of drugs leads to their frequent occurrence in the environment, which poses a risk not only because it can lead to the proliferation of antibiotic-resistant bacterial strains, but also because it can threaten human health. Therefore, this work focuses on the issue of the occurrence and behaviour of two antibiotics in soil, specifically tetracycline and sulfamethoxazole. Due to the presence of soil organic matter, it is believed that the soil can partially prevent the spread of drugs that enter the environment. The diploma thesis involved conducting diffusion experiments in soil columns under real conditions to determine the effective diffusion coefficients of tetracycline and sulfamethoxazole in three extraction agents. Sorption and desorption processes were also conducted in the individual layers of the soil column. The concentrations of the leachates were measured using UVVis spectrometry and liquid chromatography, while FTIR spectrometry was used to confirm the presence of the bound drug in the soil column.
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Micro/macro-scale investigation of the viscoelastic properties of hydrogel materials
Obrusníková, Klára ; Pekař, Miloslav (referee) ; Smilek, Jiří (advisor)
Tato diplomová diplomová práce byla zaměřená na srovnání mezi mikroreologickými metodami a makroreologickým přístupem, zejména při charakterizaci hydrogelů. Dynamický rozptyl světla, fluorescenční korelační spektroskopie a optická pinzeta byly vybrány jako mikroreologické techniky, zatímco oscilační reometrie sloužila jako makroreologický nástroj. Výsledky z dynamického rozptylu světla a fluorescenční korelační spektroskopie pro hydrogely agarózy a gellanu byly porovnány s reometrií a mezi sebou. Hodnoty viskoelastických modulů získané z mikroreologických měření byly výrazně nižší než ty získané pomocí makroreologie. Jak dynamický rozptyl světla, tak fluorescenční korelační spektroskopie nabízejí širší škálu měřitelných frekvencí než reometrie, ale obě metody mají své limitace při charakterizaci hydrogelů. U dynamického rozptylu světla dochází k odrazu světla od gelové sítě, což způsobuje šum a zhoršuje kvalitu získaných výsledků. Fluorescenční korelační spektroskopie, přes svou vyšší specifitu, vykazuje jen velmi malé rozdíly mezi vzorky různých koncentrací. Optická pinzeta je na Fakultě chemické novou metodou, a byla provedena pouze kalibrační měření. Tuhost optické pasti, důležitá hodnota pro kalibraci optické pinzety, byla stanovena pro roztoky glycerolu.
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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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Transport and mechanical properties of hybrid hydrogel networks based on gellan
Sokolínský, Jan ; Pekař, Miloslav (referee) ; Kalina, Michal (advisor)
This diploma thesis deals with the characterization of the mechanical and transport properties of gellan based hybrid hydrogels. As part of this work, the process of optimizing the preparation of gellan-poly(vinylalcohol) IPN hydrogel was first studied with an emphasis on the repeatability of the preparation due to the preservation of mechanical properties of the hydrogel. Then, both on the gellan hydrogels themselves and on selected IPN compositions, the effect of various additives was studied (specifically, the effect of the addition of a plasticizer and the effect of cross-linking by polyvalent ions). Based on these tests, a set of samples was selected, on which dynamic mechanical analysis, swelling and drying experiments were additionally measured. At the end of this diploma thesis, the transport properties of selected hydrogel compositions were also studied. The tests conducted in this thesis revealed that both the formation of a dual network and the addition of the mentioned additives allow us to influence the studied properties.
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Utilization of novel strategies of alginate crosslinking in the development of next-generation bioinoculants
Gašparová, Dominika ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
Tato diplomová práce se zaměřuje na různé strategie síťování alginátu, s cílem posunout vpřed vývoj bioinokulantů nové generace. Zkoumání různých strategií gelování se aplikuje na pět komerčně dostupných alginátů a izolovaný bakteriální alginát. Experimenty zahrnující přípravu alginátového makrogelu sloužily ke stanovení výskytu a kvality síťování, které byly analyzovány amplitudovými testy. Výběr vhodných síťovacích činidel a stanovení jejich optimálních koncentrací byly klíčovými cíli. Síťovací činidla, která byla vybrána jako vhodná pro makro- i mikroenkapsulační procesy, byly ty obsahující chlorid zinečnatý, chlorid hlinitý, chlorid barnatý, síran vápenatý a chlorid železitý. Optimální podmínky síťování byly stanoveny při teplotě 7~°C a optimální délka doby síťování v rozmezí od 30 do 120 minut. Byly provedeny komparativní analýzy šesti vybraných síťovacích činidel a to prostřednictvím analýzy mechanických vlastností skrz squeeze test, termogravimetrickou analýzou, botnáním gelu a obrazovou analýzou. Síťování barnatým síťovacím činidlem prokázalo slibné výsledky při zachování obsahu vody během rehydratace, zatímco sítování zinečnatým síťovacím činidlem dosáhlo nejvyššího obsahu sušiny. Obrazová analýza odhalila menší gelové perly tvořené síťovacími činidly s obsahem hliníku a barya, což naznačuje jejich účinnost. Mikroenkapsulované gelové perly, zejména ty síťované s baryem, prokázaly potenciál díky jejich menší velikosti. Vrcholem pokusů bylo síťování bakteriální kultury, kde hliníková a barnatá síťovací činidla dosahovala nejmenších gelových perel. Obzvlášť síťování barnatým činidlem dosahovalo nejmenších velikostí s nejmenší odchylkou, indikujíc jeho efektivitu.
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Computer modeling of diffusional transport in hydrogel
Koláček, Jakub ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
This thesis focuses on the design of models applicable to the simulation of particle motion in a viscoelastic environment in COMSOL Multiphysics. Two approaches have been chosen for the design of these models: the first is the design of a geometry representing the porous structure of hydrogels and the second is the implementation of viscoelasticity using the mathematical concept of a continuous environment. Two elementary geometrical models are presented in this work – a three-dimensional periodic lattice and a spherical volume model. Furthermore, the possibilities of implementing the generalized Langevin equation in COMSOL Multiphysics are explored by using the interface for custom partial differential equations, by adding an auxiliary dependent variable, or by defining a custom external function written in the C language. The proposed geometric models did not prove to be suitable for the simulation of viscoelastic environment. An implementation using an external function seems to be the most promising, as it offers the most customization possibilities, and its implementation reflects the theoretical foundations. The thesis also includes a custom add-in written for COMSOL Multiphysics to facilitate the evaluation of simulation data and a modified Python script to calculate the complex shear modulus from MSD data.
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Preparation and characterization of triple-IPA for the formation of vesicular systems
Novotná, Ludmila ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This thesis deals with the preparation and characterization of vesicular systems consisting of triple-IPA (ion pair amphiphile with three hydrophobic chains). For the preparation of tripleIPA were used double-chain cationic surfactants dimethyldimyristylammonium bromide (DMSAB), dimethyldipalmitylammonium bromide (DPAB), dimethyldioctadecylammonium bromide (DODAB) and dimethyldioctadecylammonium chloride (DODAC) and the singlechain anionic surfactant sodium dodecyl sulfate (SDS). Among the cationic surfactants, DMSAB and DPAB were selected to be studied more thoroughly. Triple-IPAs were prepared by mixing solutions of oppositely charged surfactants in equimolar ratio, and the resulting triple-IPA precipitate was filtered and dried. From the obtained powder, vesicular systems were prepared by rehydration and sonication. Cholesterol and cationic surfactants, from which the triple-IPAs were prepared, were used to stabilize the cationic vesicles. The properties of the vesicular systems were evaluated by dynamic and electrophoretic light scattering measurements and turbidimetry. The dispersion solutions with the addition of cholesterol were less turbid; therefore, it can be assumed that they supported the formation of cationic vesicles. The optimum cholesterol content is 20 mol. %. At this concentration, the effect of cholesterol on vesicle membrane reorganization becomes apparent and the cationic system remains monodisperse. A positive zeta potential in the stable region was determined for all prepared solutions, which was further increased by the addition of a positive surfactant. The most suitable cationic surfactant for the preparation of triple-IPA is DPAB, which is easy to work with, produces the most monodisperse systems and has the lowest measurement uncertainties.
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Hydrogels modified by amphiphilic structures
Heger, Richard ; Sedlařík, Vladimír (referee) ; Kráčalík, Milan (referee) ; Pekař, Miloslav (advisor)
The submitted dissertation deals with the influence of amphiphilic structures on hydrogel properties. Additions of various amphiphilic substances associated with the formation of highly ordered structures affect the mechanical, transport and structural properties of hydrogels. The main inspiration for this type of work was living tissue, more precisely the extracellular matrix, which is often mimicked by hydrocolloids, and its high orderliness is responsible for its unique properties. The knowledge obtained from this tissue was applied to the hydrogel systems studied in this work. Various cross-linked hydrogel matrices (physically cross-linked agarose and gelatin, ionically cross-linked alginate and chemically cross-linked mixture of polyvinyl alcohol and chitosan) were suitable representatives for this work. These hydrogel systems were modified by the addition of various amphiphilic substances. The human body’s own phospholipid, lecithin, or variously charged more classic surfactants (CTAB, SDS and Triton X-100). Experimentally, this work is divided into three areas, the study of mechanical properties using rheology, the description of transport properties via release and flow experiments using various model drugs (rhodamine 6G, eosin B, amido black 10B, methylene blue and riboflavin), and morphological characterization using SEM. The characterization of hydrogel systems was supported by other techniques used in this work, such as drying and swelling experiments or gas sorption.
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Hyaluronan micro- and nanoparticles
Mourycová, Jana ; Marián, Lehocký (referee) ; Pekař, Miloslav (advisor)
The aim of this thesis was to prepare hyaluronic acid micro- and nanoparticles based on electrostatic interactions with oppositely charged molecules. Following parameters were monitored: correlation function behavior, the particle size and zeta potential value. At the beginning, it was necessary to study the behavior of hyaluronan in solution by dynamic light scattering measurement. Micro- and nanoparticles were prepared by mixing different volume ratios of negatively charged hyaluronan and positively charged polyarginine or cetyltrimethylammonium bromide. Micro- and nanoparticles were prepared in aqueous solution as well as in 0,15 M sodium chloride solution (physiological solution). In the case of the hyaluronan solution a polydisperse character of hyaluronan was detected. It was found that the dissolution of hyaluronan in the physiological solution gives us the smaller particle size in opposite to particle size obtained from the same concentrations of hyaluronan dissolved in water. Furthermore, it was found that systems composed of hyaluronan and polyarginine create particle size of about 100 nm. Whereas systems consisting of cetyltrimethylaminoum bromide and hyaluronan form larger particles, in units of hundreds of nanometers, the particle size in physiological solution were smaller than the same systems dissolved in aqueous solution.
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