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Hyaluronan oligosacharides and their biological function
Kučerová, Barbora ; Netopilová, Miloslava (referee) ; Lázníčková, Alice (advisor)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysic and Physical Chemistry Candidate: Bc. Barbora Kučerová Supervisor: Doc. Ing. Alice Lázníčková, CSc. Title of diploma thesis: Hyaluronan oligosaccharides and their biological function In my diploma thesis I compile informations about hyaluronan oligosaccharides. Although hyaluronan is in the body found mostly in a form of long chains, we can find it also as small fragments - the oligosaccharides. The oligosaccharides embody different properties than hyaluronan polymers. Hyaluronan oligosaccharides can be synthetised, separated and analysed in laborarory in a variety of ways. Large molecules can be broken into fragments chemically, physically or enzymatically. For thein subsequent separation and purification especially chromatographic methods are used. As the most suitable methods for their final analysis electrophoretic techniques or mass spectrometry were shown. Continous metabolism of hyaluronan proceeds inside the body . Hyaluronan synthase takes a part in synthesis, and the biodegradation important for human is supplyed by several hyaluronidases. Interaction of hyaluronan with cells is mediated by several receptors. Specific receptor for hyaluronan is CD44, which can bind either large polymers and...
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Establishment of CD method for studies of amyloid-oligomerization process during Alzheimer disease
Vilímová, Jana ; Kubíček, Vladimír (referee) ; Lázníčková, Alice (advisor)
One of the main features of Alzheimer's disease (AD) are senile plaques in the brain. They are composed of β-amyloid (Aβ) peptides, which oligomerize and form fibrils during pathological conditions. Inhibition of this oligomerization process is believed to be a possible therapy to prevent the progression of AD. Circular dichroism (CD) spectrometry is an optical method which enables to follow changes in the secondary structures of proteins. Within this study a CD-method was established by which the oligomerization process of synthetic Aβ 42 was monitored. During the validation of the method several parameters as the influence of several organic solvents as Ethanol, Methanol and DMSO on the spectra and the solvent solution and concentration of Aβ 42 were assessed. Then it was investigated if the addition of proposed disintegrating substances (natural compound, fluorescence dyes and antibiotics) resulted in a delayed onset of the oligomerization process of Aβ 42. Since these substances were dissolved in different solvents, the possible effects of the solvents alone as appropriate blank values were also carried out. The oligomerization experiments were carried out at 37 řC and CD-spectra were recorded at several time points (0, 7, 24, 48 hours). In general, resulted CD-spectra showed concentration...
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Study of radiolabelling of hyaluronic acid by oxidative iodination
Dvořáčková, Eliška ; Lázníčková, Alice (advisor) ; Kubíček, Vladimír (referee)
Study of radiolabelling of hyaluronic acid by oxidative iodination Diploma paper Eliška Dvořáčková 2009 Charles University in Prague, Pharmaceutical Faculty in Hradec Králové Department of biophysics and physical chemistry In the frame of this work the literature describing characteristics and possible applications of hyaluronic acid and methods of radiolabelling of compounds with a view to the radioiodination was read up. In experimental part of the task two methods of oxidative iodination of hyaluronic acid modified by tyramine with the lowest degree of modification ( HA - TM - 1 ) was examined. Chloramine method, used as first one, turned out ineligible as due to presence of oxidative agent in the phase with hyaluronic acid a partial degradation of the sequence occurred. Another method of iodination with the use of iodogen is heterogenous reaction, oxidative agent is present as an insoluble deposit on the walls of the reaction vial. This method of radiolabelling was more moderate to hyaluronic acid. The reaction was interrupted by applying the reaction system on the column with Sephadex G - 50 wherby the labelled compound was separated from the aggressive reactive environment. It turned out that this way of refinement of the labelled product does not eliminate the rests of the radioactive iodide....
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Stability studies of monoclonal antibody nimotuzumab modified with chelator DOTA radiolabelled with lutetium-177 ([177Lu]Lu-hR3(p-SCN-Bn)DOTA)
Petřík, Michal ; Lázníčková, Alice (advisor) ; Kubíček, Vladimír (referee)
The higher expressivity of the gene for the receptor of epidermal growth factor (EGF) and consequently higher number of receptors (EGFR) on the surface of cells is closely linked with the tumour proliferation of these cells and with poor prognosis for patients. For such reason the EGFR became one of the main objectives in the therapy of tumours. Nimotuzumab (also known as h-R3) is humanized monoclonal antibody which blocks receptor for epidermal growth factor. The object of this thesis was to determinate radiochemical purity of this antibody conjugated with bifunctional chelator DOTA and marked by luthecium, possibly purification of this antibody and consequently determination of its stability in different environments. In acetate buffer pH = 7, in an environment of the surplus of competitive ligand EDTA and in plasma of rats. Adequate analytic methods were used for these tasks: high-efficiency liquid chromatography, gel permeation chromatography and chromatography on thin layer. The results of the thesis demonstrated that this way of modification for further radiolabelling is not appropriate for large protein structures such as Nimotuzumab
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Use of the polarized light to study interactions of substances in solutions II
Otradovská, Zuzana ; Kubíček, Vladimír (advisor) ; Lázníčková, Alice (referee)
This rigorous thesis deals with a study of the interaction between sodium lauryl sulphate and organic dye neutral red utilizing fluorescence spectroscopy. The aim of this work was to verify interactions of tenside and organic dye in their solutions, which were indicated in my diploma thesis, by a further method. The work serves as the introductory basis for a possible utilization of fluorescence measurement with polarized light in biomedical and gerontological research. A study of the solutions gave very interesting results that indicate the formation of aggregates. But the question, if the aggregates are of highly arranged type or disarranged type, remains.
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Influence of Electrolytes on CMC
Šmerdová, Jana ; Kubíček, Vladimír (advisor) ; Lázníčková, Alice (referee)
Souhrn V této diplomové práci bylo použito UV-VIS absorpční spektrofotometrie k měření CMC katonických tenzidů a vlivu elektrolytů na CMC. Dosažené výsledky potvrzují očekávaný trend poklesu hodnot CMC tenzidů se stoupající iontovou silou roztoků. Použitá modifikace metody UV-VIS absorpční spektrofotometrie pro stanovení CMC byla rovněž úspěšná.
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