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Adsorption of microbial cells on model biological surfaces
Mikušová, Janka ; Krouská, Jitka (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is creating sorption surface suitable for studying adsorption bacterial strain Lactobacillus rhamnosus, as representative probiotic bacterial strain. Included in the experimental part of this thesis was the process of creating sorption surface, according to the designed and especially optimized method. Optimization method comprised of preparing sorbent with sorption surface including mucin, on which was the bacterial strain Lactobacillus rhamnosus bound to, using specific interaction. UV-VIS spectrophotometry was utilized in the optimization of the sorption surface preparation for monitoring losses of mucin in solution that happens during the bondation on used substrate. Designed sorption surfaces were subjected to various physical-chemical analyses, which roles were displaying and characterisation of surface and confirming the presence of mucin. One of the major components, in the process of observing and characterization of changes in surface structure of the sorption surface, was infrared spectroscopy with Fourier transformation. Scanning electron microscopy was used for more detailed observation of the surface structure, created by adhesive interactions, which participate in probiotic bacteria adhesion. Finally, the created surfaces were used in pilot experiment, during which, the kinetics in growing bacterial culture with and without the presence of sorbent using microcalorimetry were observed. Designed sorbent appears to be a suitable mediator for detailed characterisation of bacterial adhesion, which is the first step in creating bacterial biofilm.
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Mucin hydrogels - artificial models of native mucus systems
Mikušová, Janka ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
The scope of this masters thesis is the preparation of a model mucin system and its utilization as an artificial model of the native mucus system. The creation of this model system, according to several designed methods was a part of experimental part of the thesis. The preparation of mucin system comprised of physical and chemical methods of hydrogel formation, screening and characterisation of the various physical conditions of the mucin properties on its molecular level, and the preparation of sorbent with sorption surface containing mucin. Methods of light scattering, namely dynamic light scattering (DLS), used for mucin particles size change monitoring, and electroforetic light scattering (ELS), used for Zeta potential change monitoring, were used for the screening of the impact of physical factors on the properties of mucin.For the characterisation of impact of the temperature on changes in mucin sctructure was, apart from monitoring of light scattering, used also a diferential scanning calorimetry (DSC), which registered temperature value, at which mucin thermal denaturation occurs. In the next part of the thesis we subdued the created sorption surfaces to various physical-chemical analyses, which task is the characterisation and projection of surface and confirmation of mucin presence.Substancial part in monitoring and characterisation of changes in surface sctructure of sorption surface was accomplished by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscophy (SEM) was used for the final, more detailed, projection of the mucin enriched, sorbent surface structure. Suggested methods of mucin hydrogel, didnt prove sufficient results for the possibility of application of hydrogel as a artificial model of real mucus system, but the sorbent application was indicated as a suitable alternative and an instrument for the further mucin behaviour research and possibly subsequent bacterial adhesion, which represents the first step in the formation of the bacterial biofilm.
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Characterization of adsorption properties of probiotic bacteria
Černá, Klára ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of the diploma thesis is to characterize the adsorption properties of probiotic bacteria. Characterization was performed using of designed and optimized spectrophotometric method of determination of adhesion of probiotic bacteria to mucin as the main viscoelastic component of mucus. In order to provide a more detailed description of the adhesion interactions involved in the adhesion of probiotic bacteria to mucin, the viability of selected probiotic bacterial strains Lactobacillus rhamnosus CCM 1825, Lactobacillus plantarum CCM 7039, Lactobacillus delbrueckii subsp. bulgaricus CCM 7190, Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 4798, Bifidobacterium breve CCM 7825, Bacillus coagulans CCM 2658 and a potential probiotic strain of Lactobacillus zeae CCM 7069. Moreover, bacterial adhesion to carbohydrates was also determined for all the bacteria tested. The last proposed and optimized technique was dynamic and electrophoretic light scattering characterizing surface properties such as -potential, average size distribution and isoelectric point. From this method, information was obtained on aggregation of bacterial cells and on electrostatic interactions. The combination of these methods was used as a complex tool to characterize adhesion of the tested bacterial cultures as a very specific, sensitive and key parameter of a successful probiotic microorganism that is influence by the multiple effects.
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Clinico-pathological characteristics of mucin-producing colorectal carcinoma
Sojka, Ladislav ; Levý, Miroslav (advisor) ; Boublíková, Ludmila (referee) ; Klos, Dušan (referee)
Colorectal carcinoma (CRC) is one of the most common malignancies worldwide with persistently high mortality. As a consequence, high demands are placed on research into this disease. The aim of this study was to evaluate the epidemiology of colorectal carcinoma in the Czech Republic in comparison with worldwide data. We focused on the evaluation of the latest findings in screening and diagnosis, investigated prognostic factors, and also focused on the therapeutic options of CRC. Furthermore, mucin-producing carcinomas are described in detail and mucin itself was confirmed as a risk factor for the prognosis of CRC patients. The experimental part of the study focused on gene mucin MUC13 and the MUC13 - miR-4647 axis, which is associated with adverse prognosis in CRC patients. Samples of CRC and samples of adjacent non-malignant mucosa tissue were used. The samples were collected from patients who underwent surgery at the General University Hospital in Prague, at the Thomayer University Hospital in Prague and at the University Hospital in Pilsen (Czech Republic). The collection of samples took place between 2011 and 2015. The total of 187 samples were collected from patients with sporadic colorectal cancer. The subjects of the study all provided information on their lifestyle habits, body mass index...
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Adhesive and rheological properties of chitosan-based mixtures
Korpasová, Marie ; Šnejdrová, Eva (advisor) ; Mužíková, Jitka (referee)
CHARLES UNIVERSITY IN PRAGUE Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical technology Name and surname: Marie Korpasová Title of diploma thesis: Adhesive and rheological properties of mixtures based on chitosan Supervisor: PharmDr. Eva Šnejdrová, Ph.D. Diploma thesis deals with evaluation of rheological and adhesive properties of compounds for matrix tablets formulation, based on Chitosan with addition of retardant compound. The retardant compound consists of Sodium Alginate and Hypromellose in concentrations of 30 %, 40 % and 50 %. Theoretical part describes Chitosan, Sodium Alginate and Hypromellose. As next is division of matrix tablets. Theoretically is also described evaluation of rotational, oscillational and adhesive testing which was used in practical part. From viscosity curves can be seen, that viscosity is decreasing with increasing stress. Viscosity increases with increasing concentration of Sodium Alginate and Hypromellose. Oscillational testing is better for gel characterization, because it provides lower stress on test samples. Samples of given compound were prepared by hydrating with phosphate buffer of pH 6,8. Coefficient of consistency K and index flow rate n characterize rheological behaviour of gels. Viscoelastic compounds behave like solids with plastic...
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Mucin hydrogels - artificial models of native mucus systems
Mikušová, Janka ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
The scope of this masters thesis is the preparation of a model mucin system and its utilization as an artificial model of the native mucus system. The creation of this model system, according to several designed methods was a part of experimental part of the thesis. The preparation of mucin system comprised of physical and chemical methods of hydrogel formation, screening and characterisation of the various physical conditions of the mucin properties on its molecular level, and the preparation of sorbent with sorption surface containing mucin. Methods of light scattering, namely dynamic light scattering (DLS), used for mucin particles size change monitoring, and electroforetic light scattering (ELS), used for Zeta potential change monitoring, were used for the screening of the impact of physical factors on the properties of mucin.For the characterisation of impact of the temperature on changes in mucin sctructure was, apart from monitoring of light scattering, used also a diferential scanning calorimetry (DSC), which registered temperature value, at which mucin thermal denaturation occurs. In the next part of the thesis we subdued the created sorption surfaces to various physical-chemical analyses, which task is the characterisation and projection of surface and confirmation of mucin presence.Substancial part in monitoring and characterisation of changes in surface sctructure of sorption surface was accomplished by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscophy (SEM) was used for the final, more detailed, projection of the mucin enriched, sorbent surface structure. Suggested methods of mucin hydrogel, didnt prove sufficient results for the possibility of application of hydrogel as a artificial model of real mucus system, but the sorbent application was indicated as a suitable alternative and an instrument for the further mucin behaviour research and possibly subsequent bacterial adhesion, which represents the first step in the formation of the bacterial biofilm.
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Adhesive and rheological properties of chitosan-based mixtures
Korpasová, Marie ; Šnejdrová, Eva (advisor) ; Mužíková, Jitka (referee)
CHARLES UNIVERSITY IN PRAGUE Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical technology Name and surname: Marie Korpasová Title of diploma thesis: Adhesive and rheological properties of mixtures based on chitosan Supervisor: PharmDr. Eva Šnejdrová, Ph.D. Diploma thesis deals with evaluation of rheological and adhesive properties of compounds for matrix tablets formulation, based on Chitosan with addition of retardant compound. The retardant compound consists of Sodium Alginate and Hypromellose in concentrations of 30 %, 40 % and 50 %. Theoretical part describes Chitosan, Sodium Alginate and Hypromellose. As next is division of matrix tablets. Theoretically is also described evaluation of rotational, oscillational and adhesive testing which was used in practical part. From viscosity curves can be seen, that viscosity is decreasing with increasing stress. Viscosity increases with increasing concentration of Sodium Alginate and Hypromellose. Oscillational testing is better for gel characterization, because it provides lower stress on test samples. Samples of given compound were prepared by hydrating with phosphate buffer of pH 6,8. Coefficient of consistency K and index flow rate n characterize rheological behaviour of gels. Viscoelastic compounds behave like solids with plastic...
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Rheological behaviour of mixtures for freeze-drying
Vavřich, Dominik ; Šnejdrová, Eva (advisor) ; Šklubalová, Zdeňka (referee)
CHARLES UNIVERSITY Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Name of author: Dominik Vavřich Title of diploma thesis: Rheological behaviour of mixtures for freeze-drying Supervisor: PharmDr. Eva Šnejdrová, Ph.D. The diploma thesis deals with the evaluation of rheological properties of aqueous dispersions of fish gelatin and carrageenans intended for lyophilization. Theoretical part of this work is dedicated to the characterization of fish gelatin and carrageenans which were used for preparation of the mixtures in experimental part of this work. Characterization of an oral cavity from an application of medicaments point of view and an orally disintegrating tablets of the medicine with a focus on the lyophilized tablets are also presented. Mixtures of the fish gelatin and the carrageenans were prepared in the experimental part of the work. Their rheological behaviour employing an absolute rotational rheometer Kinexus Pro+ was measured and evaluated. Also, the testing of the rheological characteristics of these materials transformed into the freeze-dried tablets was performed. Nonlinear viscous curves were obtained, and they were mathematically evaluated by the Power law model. Pseudoplastic behaviour of the mixtures can be derived from the results of acquired...
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Adsorption of microbial cells on model biological surfaces
Mikušová, Janka ; Krouská, Jitka (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is creating sorption surface suitable for studying adsorption bacterial strain Lactobacillus rhamnosus, as representative probiotic bacterial strain. Included in the experimental part of this thesis was the process of creating sorption surface, according to the designed and especially optimized method. Optimization method comprised of preparing sorbent with sorption surface including mucin, on which was the bacterial strain Lactobacillus rhamnosus bound to, using specific interaction. UV-VIS spectrophotometry was utilized in the optimization of the sorption surface preparation for monitoring losses of mucin in solution that happens during the bondation on used substrate. Designed sorption surfaces were subjected to various physical-chemical analyses, which roles were displaying and characterisation of surface and confirming the presence of mucin. One of the major components, in the process of observing and characterization of changes in surface structure of the sorption surface, was infrared spectroscopy with Fourier transformation. Scanning electron microscopy was used for more detailed observation of the surface structure, created by adhesive interactions, which participate in probiotic bacteria adhesion. Finally, the created surfaces were used in pilot experiment, during which, the kinetics in growing bacterial culture with and without the presence of sorbent using microcalorimetry were observed. Designed sorbent appears to be a suitable mediator for detailed characterisation of bacterial adhesion, which is the first step in creating bacterial biofilm.
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Characterization of adsorption properties of probiotic bacteria
Černá, Klára ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of the diploma thesis is to characterize the adsorption properties of probiotic bacteria. Characterization was performed using of designed and optimized spectrophotometric method of determination of adhesion of probiotic bacteria to mucin as the main viscoelastic component of mucus. In order to provide a more detailed description of the adhesion interactions involved in the adhesion of probiotic bacteria to mucin, the viability of selected probiotic bacterial strains Lactobacillus rhamnosus CCM 1825, Lactobacillus plantarum CCM 7039, Lactobacillus delbrueckii subsp. bulgaricus CCM 7190, Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 4798, Bifidobacterium breve CCM 7825, Bacillus coagulans CCM 2658 and a potential probiotic strain of Lactobacillus zeae CCM 7069. Moreover, bacterial adhesion to carbohydrates was also determined for all the bacteria tested. The last proposed and optimized technique was dynamic and electrophoretic light scattering characterizing surface properties such as -potential, average size distribution and isoelectric point. From this method, information was obtained on aggregation of bacterial cells and on electrostatic interactions. The combination of these methods was used as a complex tool to characterize adhesion of the tested bacterial cultures as a very specific, sensitive and key parameter of a successful probiotic microorganism that is influence by the multiple effects.
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