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Cationic fluorescence probes in a polyanion-surfactant system.
Mondek, Jakub ; Šmejkalová, Daniela (referee) ; Pekař, Miloslav (advisor)
In this bachelor thesis were studied properties of aqueous solutions of hyaluronan and celytrimethylamonnium bromid with help of acridine orange as fluorescence probe. At first, this thesis was focused on interactions of acridine orange and hyaluronan. Emision and excitation spectra of acridine orange were observed. Result of these measurement was bonding of acridine orange dimer to carboxylic group of hyaluronan. Next studied interaction was interaction of cetyltrimethyl amonium bromid with acridine orange – hyaluronan system. Result of these interaction was, that molecules of cationactive surfactant pushed away dimers of acridine orange and took place of dimer of acridine orange on carboxylic group of hyaluronan. Dimer of acridine orange was changed into monomers. The reason why this thesis was focused on study of aqueus solutions of hyaluronan with cationactive surfactant is benefit of hyaluronan – surfactant system in research of transport system for target distribution of drugs.
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Stability evaluation of cosmetic dispersions using centrifugal analyser
Procházková, Michaela ; Krouská, Jitka (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis is focused on the evaluation of stability of cosmetics emulsions prepared from ingredients commonly available in laboratory and compared to the stability of commercially available body lotions. In the theoretical part cosmetics emulsion and methods used to stability measurements were described. Experimental part of this work at first describes the preparation of the emulsions. Furthermore, the stability of prepared emulsion was compared to commercially available ones using the centrifugal analyser LUMiSizer. In conclusion, the test results obtained from analyser LUMiSizer have showed the stability of the commercial emulsions and two prepared samples.
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Incorporation of microbial cells in hydrogel carriers
Orišková, Sofia ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
The presented diploma thesis focuses on the use of plant growth promoting bacteria as an ecological alternative to conventional fertilizers. The incorporation of bacterial cells into hydrogel carriers is already a well-studied topic, but due to its disadvantages it has not yet found wider application in agriculture. This work offers a novel concept of encapsulating bacteria by gelation directly from the culture. This is achieved by crosslinking the bacterial alginate produced by the model microorganism Azotobacter vinelandii. Since this process was not described before, first its optimization was needed. Alginate production was determined gravimetrically, and its parameters were further characterized using available analytical methods – infrared spectroscopy to monitor structural parameters (monomer composition and the extent of acetylation), dynamic light scattering to characterize the size distribution and AF4-MALS-dRI to obtain the molecular weight. Bacterial PHB production was also investigated using gas chromatography and infrared spectroscopy. The second part of the work is focused on the optimization of the gelling process using bacterial alginate from the culture and CaCl2 as a crosslinking agent. Rheological experiments were used as a tool in understanding the viscoelastic properties of the prepared gels. Gelation was demonstrated within the first day after inoculation. Maximum production of alginate (1,9 ± 0,3) g/l was reached on the fourth day after inoculation. It was found that the addition of 5 g/l of calcium carbonate promotes the production of alginate. Nevertheless, further addition of CaCO3 (30 g/l) showed adverse effects on the molecular weight and is therefore not recommended. Production of PHB was confirmed by both FTIR and GC measurements, with a maximum yield of (23 ± 3) % CDW. Rheological testing confirmed that the product of the crosslinking was a gel. It was found that the crosslinker concentration plays an important role at time 0 min of the gelation, forming a denser network in the structure and causing higher rigidity. Using the highest studied concentration of CaCl2, the critical strain reached values of (5,0 ± 0,7) %. Finally, the incorporation of bacterial cells into the hydrogel was confirmed using fluorescence microscope.
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Interaction Between Hyaluronan and Amphiphilic Molecules
Venerová, Tereza ; Burget, Ladislav (referee) ; Čeppan, Michal (referee) ; Pekař, Miloslav (advisor)
This work is focused on interaction between hyaluronan (Hya) and amphiphilic molecules. Using fluorescent probes method were carried out screening of the interaction of various surfactants with Hya. For further examination was chosen cetyltrimethylammonium bromide (CTAB). Interactions in this system have also been studied using fluorescence correlation spectroscopy (FCS), and additionally were performed initial tests of the cytotoxicity of the system. As an alternative CTAB were also used hydrophobically modified amino acids. Results showed their self-aggregation, but their interactions with hylauronanem was not as strong as with CTAB. The interaction of gel systems Hya-CTAB have also been studied, and was found to contain a hydrophobic domain capable solubilize hydrophobic substance. Thermogravimetric have been established that are able to contain up to 98% wt. water and rheologically were characterized their mechanical properties. These are tunable due to molecular weight (MW) of the hyaluronate. With increasing value of MW increases mechanical strength of the gel and decreases its fluidity.
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Microrheology of Biocolloids
Hnyluchová, Zuzana ; Omelka, Ladislav (referee) ; Kráčalík, Milan (referee) ; Pekař, Miloslav (advisor)
The main aim of the doctoral thesis was study of passive microrheological techniques as advanced methods for characterisation of viscoelastic properties of soft material. These techniques are able to provide certain advantaged or additional information on classical rheology. Several techniques from the wide range of passive microrheological group such as one particle video – microrhelogy with IDL data procesing, microrheology based on light scattering – DLS or FCS microrheology have been applied in practice during my PhD studies. New Matlab script has been also invented to simply obtain information about viscosity of low volume samples. Aplicability of techniques were studied on several various bicolloids such as solutions or gels of hyaluronic acid, on study of properties at the interfaces or on gelation proces studies of agarose. Obtained data were compared classical rheology results. Suitability of each technique for investigated systems or appropriate reccomendations for further measurements were didcussed based on obtained data. Main advantages and limites of passive microrheology techniques were also described in comparison wih classical rheology method.
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Rheology of Hyaluronan Solutions
Bilerová, Helena ; Porkorný,, Marek (referee) ; Lehocký,, Marián (referee) ; Pekař, Miloslav (advisor)
Předložená práce se zabývá reologickými vlastnostmi roztoků hyaluronanu. Studuje chování hyaluronanových roztoků v prostředí několika rozpouštědel, dále se zabývá vlivem měnícího se pH a rostoucí teploty na viskozitu roztoků. V neposlední řadě je zkoumána správná příprava roztoků, jejich časová stabilita a tvorba agregátů.Všechny experimenty vycházejí z praktického upotřebení, např. při výrobě hyaluronanu, jeho sterilizaci a následném skladování roztoků.
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Dialysis study of hyaluronan-surfactant interactions
Šejnohová, Michaela ; Venerová, Tereza (referee) ; Pekař, Miloslav (advisor)
This diploma thesis is concentrated on the interactions between polyelectrolyte (hyaluronan) and cationic surfactant (CTAB). The experiments were performed in an aqueous solution and in an environment of physiological ionic strength (0,15mmoldm-3 NaCl). The determination of the surfactant concentration in solutions was based on the formation of colored complexes of CTAB and picric acid in chloroform. The concentrations of surfactant were measured by UV-VIS spectroscopy. The stability of CTAB+HyA was examined by a dialysis method. The results showed that, regardless of the environment, the presence of HyA in solution reduces the number of free molecules of CTAB which can be determined in the sample. It has been proved that there is an interaction between HyA and surfactant and that CTAB has greater affinity for HyA then for the picric acid. The stability of CTAB+HyA was determined by dialysis of 120 hours. After that time, the concentrations of the retentate and permeate were settled. The results showed that in the membrane remains a certain amount of CTAB bounded to hyaluronan. The system can be suitable for the preparation of targeted carriers of biologically active substances.
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Correlation of Thermoanalytical Data with Primary Properties of Humified Substrata
Kislinger, Jiří ; Šimon, Peter (referee) ; Tesařová, Marta (referee) ; Schaumann, Gabriele (referee) ; Pekař, Miloslav (advisor)
Tato dizertační práce je napsána jako termoanalytická studie humifikovaných substrátů, tj. huminových látek a půdních vzorků, získaných z několika různých zdrojů tak, aby byl pokryt co nejširší rozsah primárních charakteristik. Práce hledá lineární korelace (s využitím Pearsonova korelačního koeficientu) mezi kinetikou termo-oxidační degradace (sledovanou pomocí termogravimetrie (TGA) a vyjádřenou konverzním časem stanoveným neizotermickou izokonverzní metodou) a standardními charakteristikami, jako je množství aromatického a karboxylového uhlíku přítomného v huminových látkách stanoveného spektroskopií nukleární magnetické rezonance (NMR). Hlavním cílem této práce je objasnit proces termo-oxidační degradace humifikované části přírodní organické hmoty a hledat závislosti mezi primární a sekundární strukturou huminových látek. Dalším úkolem je nalezení korelace mezi termogravimetrickým úbytkem hmotnosti půdních vzorků a jejich mikrobiální stabilitou měřenou v laboratorních experimentech jakožto půdní respirace (tedy uvolňování oxidu uhličitého). Účelem je zjistit, zdali vzájemné propojení vůbec existuje, a objasnit jaké druhy procesů, které provázejí půdní respiraci, je možné jednoduše odhalit pomocí TGA. Výsledky práce skýtají detailnější pohled na termickou analýzu půdních vzorků a jiných humifikovaných materiálů. Hlavním přínosem této práce je prezentace vyskytujících se aplikací termické analýzy pro charakterizaci půdní organické hmoty a dále také příspěvek k rozvoji aplikací technik termické analýzy v půdní vědě.
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Energy transfer and hydrophobic domains in colloidal systems
Kučerová, Petra ; Čeppan, Michal (referee) ; Pekař, Miloslav (advisor)
In this thesis resonance energy transfer between perylene and fluorescein in tetradecyltrimethylammonium bromide (TTAB) solution was studied. The influence of addition of native or modified sodium hyaluronate on resonance energy transfer was also investigated. The addition of native sodium hyaluronate supports energy transfer at lower fluorescein concentrations and the addition of modified sodium hyaluronate influences the maximal value of energy transfer effectivity. Strong influence on the critical micelle concentration (CMC) values with the addition of sodium hyaluronate during CMC of TTAB determination was investigated. The data indicates not only micelles formation, but also formation of aggregates of sodium hyaluronate with TTAB. Aggregation numbers of TTAB with addition of native and modified sodium hyaluronate by the quenching of pyrene by cetylpyridinium chloride (CPC) was investigated. The addition of sodium hyaluronate into the solution of TTAB changes the average mean aggregation number. Solute exchange between micelles of TTAB and the influence of addition of modified sodium hyaluronate on this exchange was also investigated. No solute exchange between micelles in TTAB and in TTAB with added modified sodium hyaluronate was discovered during this experiment.
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Influence of selected active compounds for properties of gel and emulsion cosmetics
Gardoňová, Lenka ; Pekař, Miloslav (referee) ; Zemanová, Jana (advisor)
This master‘s thesis studies problems of chosen active substances in cosmetic emulsions and gels. There is their main characteristics – distribution and properties of cosmetic emulsions and gels, preparation of the active substances, effects on the skin and conditions, which could them influence. In experimental part, preparation of gels and emulsions bases was described which were than enriched by active substances - evening primrose oil, hemp oil, Salvia officinalis extract, Symphytum officinale extract and sea salt. Properties of these cosmetic gels and emulsions bases with the addition of active substances depend on temperature and method of storage, amount of light exposure, method of preparation and another conditions. Action of these factors can cause changes of the stability, thereby the cosmetics effects on the skin can be lost. On the bases of performed rheology measurements (oscillation and flow measurements), pH measurements and sensory analysis were determined, if the properties of examined samples were changed. The measurement was performed in the certain time intervals and the properties were evaluated like at laboratory temperature 23±2 °C, so after exposure to temperature regime 37 °C in thermoregulator. On the basis of performed measurements, there was determined, that gel base with addition of Symphytum officinale extract had the most stability properties.
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