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Study of expression of transferrin receptors (TfR1) and their utilization in nanomedicine
Krausová, Kateřina ; Fohlerová, Zdenka (referee) ; Heger,, Zbyněk (advisor)
Bachelor thesis deals with the expression of the transferrin receptor (TfR1) and its use in nanomedicine. During the last decade, nanotechnology emerged as one of the central milestones in connecting all scientific and technological disciplines. Nanomedicine already demonstrated efficacy not only in animal models of cancer but also in clinical practice. The theoretical part is not only aimed at cancer of the human population, but also at the possibilities of targeted drug delivery into the tumor tissue, which greatly reduces the otherwise serious side effects of conventional treatment – systemic toxicity. The practical part is focused on optimization for studying the expression of the transferrin receptor, a protein overexpressed by neoplastic cells aiming to enrich the higher metabolic needs of tumor cells. The optimal conditions were as follows: lysate of 50 000 cells applied with nonreducing nondenaturing buffer and the concentration of the primary antibody of 1.0 𝜇g/ml. Different levels of TfR1 expression were detected, depending on the type of tumor cells. The cell lines of neuroblastoma, prostate cancer (occurence in every 7th man) and breast cancer (occurence in every 8th woman) were selected for the next experiments. Via this transferrin receptor, apoferritin, which is a protein storaging iron ions in many organisms, can be internalized into cells. Artificially, the internal cavity of apoferritin may be used for encapsulation and transport of any molecules. In the case of this bachelor thesis, the apoferritin was used for delivery of doxorubicin. Doxorubicin has been used for cancer treatment for more than 30 years; however, its administered dose is limited by its high toxicity. This can be reduced by its encapsulation in a suitable vector for targeted transport to the tumor cells only. Apoferritin could serve as such suitable vector. In this thesis, the suitable usage of apoferritin as a nanocarrier for chemotherapeutic delivery was confirmed. Its molecule size of 10-12 nm allows it to employ the effect of increased permeability and retention. At the same time, this size makes it possible to avoid renal clearance. The properties of encapsulated doxorubicin are not affected by apoferritin, thus preserving its toxicity for cells with a high level of TfR1 expression (30% growth inhibition of these cells after 24 h of treatment).
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The proteomic study of abiotic stress of plants.
Barabášová, Kamila ; Podlipná, Radka (advisor) ; Smrček, Stanislav (referee)
Keywords: Arabidopsis thaliana, phytoremediation, abiotic stress, ibuprofene, doxorubicin, two-dimensional electrophoresis Nowadays, develop of the pharmaceutical industry is very fast. Reason of this trend is ever-increasing number of diseases, lifestyle and still increasing demand for the drugs. With this trend growing interest in the analysis of the residues of pharmaceuticals in the environment which is result of incomplete wastewater treatment. This diploma thesis is studying effect of cytostatic drugs, specifically doxorubicin and one of the most widely used analgesics - ibuprofen, at the proteome level of the model plant Arabidopsis thaliana. Proteins isolated from plants exposed to the drugs were separated by two-dimensional electrophoresis. Comparing of protein maps by PDQest program (Bio-Rad, USA) was found several proteins whose expression was affected by the presence of drugs in the culture medium. Selected proteins were identified by LC - MS / MS.
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Nanomaterials as a platform for the doxorubicin transport - Fluorescence imaging
Blažková, Iva
Doxorubicin (DOX) is an important anthracycline antineoplastic drug used to treat a variety of cancers. As well as other antineoplastic agents, DOX has negative side effects; the most serious one is cardiotoxicity. Conjugation of DOX with nanomaterials could help to reduce its adverse effects. The presented thesis entitled ,,Nanomaterials as a platform for the DOX transport -- Fluorescence imaging" is primarily focused on the study of DOX interaction with biomolecules such as amino acids and albumin. The interaction was analysed by fluorescence spectrometry and by other analytical methods and significant interaction with amino acids as well as with protein albumin was detected. Next work was focused on the utilization of nanoparticles in DOX transport. DOX was conjugated with fullerenes or encapsulated in liposomes and properties of these nanoconstructs, in vitro and in vivo, were analysed. Both of these nanomaterials seemed to be good nanotransporters for DOX targeted delivery.
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Study of DNA-doxorubicine interaction by electrochemical methods on nanostructured electrodes
Přibyl, Jakub ; Fohlerová, Zdenka (referee) ; Kynclová, Hana (advisor)
Cancer diseases are one of the leading causes of mortality worldwide, for this reason, great attention anticancer drugs. Doxorubicin falls into the category of the most widely used cancer drugs. The method of electrochemical impedance spectroscopy were investigated properties of golds electrodes without deposit nanoparticles and electrodes with deposit gold nanoparticles. Using voltammetric measurements we detected doxorubicin and investigated the interaction of doxorubicin with oligonucleotides on gold electrodes without nanoparticles and electrodes modified with gold nanoparticles.
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Raman spectroscopy of biologically active species and antitumor drugs
Třeštíková, Liběna ; Vetterl,, Vladimír (referee) ; Vrána, Oldřich (advisor)
SERRS spektra of biological materials are very komplex, because they consist of signals from all molecules present in cells. In this text are presented SERRS spektra of antitumor drugs and its komplex with DNA. Experimental are rated on doxorubicin and another antitumor druha and on study of theirs potential by treatment for tumors. Doxorubicin belong to clase antracycline antibiotics and is used for stop of tumor cells reproduction. Scientists found still new ways, new drugs. SERRS is one of possibilities for study of this drugs and theirs interaction with DNA.
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