|
Exprese transgenu bphC u ječmene
Sedlářová, Kateřina
Polychlorinated biphenyls are toxic organic compounds, characterized by a low degradability in the environment and accumulation in the food chain. This is the reason, why decontamination of soils polluted with such substances is very important issue for today's society. Decontamination of areas infected by PCBs is very complicated due to its physical and chemical characteristics. Appropriate decontamination method is phytoremediation, which is used for removing polychlorinated biphenyls by natural processes of plants. Using the transformation of plants can increase the efficiency of phytoremediation. The thesis has been investigated in expression of remediation gene bphC/His at T2 generation of spring barley (Hordeum vulgare L.), variety Golden Promise at the DNA, RNA and protein levels. Expression of the transgene was detected using PCR, real-time PCR and Western blot. To obtain dihaploid homozygous plants of spring barley, variety Golden Promise containing a phytoremediation gene bphC/His was used anther cultures technique. In the thesis were choosed plants from the T2 generation, showing the presence of the inserted gene to test the ability of transgenic plants to accumulate polychlorinated biphenyls.
|
|
Charge transport in DNA
Drevený, Lukáš ; Mravec, Filip (referee) ; Vala, Martin (advisor)
In this bachelor thesis were investigated charge transport (CT) through a duplexes of DNA by stationary fluorescence spectroscopy, with a fluorescent analogue of adenine - 2-aminopurine (Ap), as fluorescent probe. The aim of study was an influence of changes in the structure of duplexes DNA:DNA or DNA:RNA and effect of the presence of Hg(II) in double-stranded DNA:DNA containing a mutation thymine-thymine in the chain, in the efficiency of charge transfer along the DNA strand. Firstly, in the experimental part was investigated a efficiency of incorporation of Ap into the DNA strand from an excitation spectra and secondly, from the integrals of the emission spectras the quenched factor was calculated. It was found that the Ap can be better incorporated into the chain of DNA:RNA, which is not as rigid as a string of DNA:DNA and thus may be better accepted by the nearby bases in the chain; furthermore, it was found that the photo-induced charge transfer (PET) through the DNA strand sharply drops after exceeding a temperature corresponding with melting point of DNA chain, at which the string of DNA is unraveling. It was also found that the presence of Hg(II) in the chain of DNA:DNA does not adversely affect a charge transfer, but on the contrary, the string is after integration of Hg(II) into the chain more stable and the melting point is shifted to higher temperatures. In this thesis it was proved that photoinduced charge transfer is very sensitive process and could be used for detection of even tiny changes in the structure of nucleic acids.
|
|
The potential of deep seabed polymetallic nodules
Tormová, Lucia ; Dvořák, Antonín (advisor) ; Hadrabová, Alena (referee)
The deep seabed polymetallic nodules are one the major types of deep-sea ore resources. Oceanic deposits of these raw materials represent a potential source of numerous metals that are in short supply on the land-base deposits at the nearest future. The process of geological survey to the deep ocean floor is considered to be an onset of a quantitatively new stage in the process of mineral resource extraction. Continuation of the on-going work on polymetallic nodule deposits within the Clarion-Clipperton Zone is regarded as a serious task to be undertaken over a period until the year 2025, with a due consideration to advance in designing of an efficient mining unit and application of modern processing technologies for extraction of major metals : manganese, nickel, cobalt and copper. A prerequisite of a rational management of oceanic resources is the preservation of natural oceanic environment. Considered is the extraction's profitability of a concrete deposit within the Clarion-Clipperton Zone, which is the exploration area of the IOM, with annual production scale of 2.2 Mt of wet nodules for the year 2025.
|
|
Magnetic Nanoparticles for Targeted Delivery System of Plasmid into HEK293 cells
Bílek, Ondřej ; Svoboda, Ondřej (referee) ; Fohlerová, Zdenka (advisor)
This bachelor’s thesis deals with using of magnetic nanoparticles, SPIO nanoparticles and nanoparticles of commercial product MATra for transfection of ASAP plasmid into HEK293 cells and subsequent optimization of the process. The experimental part is enclosed with theorethical part that summarizes findings from the fields of synthesis of magnetic nanoparticles and their biomedical applications, theory dealing with possibilities of the insertion of foreign DNA or plasmids into the cell and of its efficiency evaluation.
|
|
Underpricing and the Long-Run Underperformance of IPOs
Pindroch, Michal ; Musílek, Petr (advisor) ; Witzany, Jiří (referee)
When companies go public, the shares they sell tend to be underpriced, and thus exhibit a significant price jump on the first day of trading. As a result, IPO investors materialize significant first-day returns. In the long-run, however, relative to some benchmark, investors appear to lose out by continuing to hold the stocks of firms that have recently gone public. These IPO phenomena are subject of the following study. The thesis addresses two main objectives. First, it systematically surveys relevant empirical evidence and theories that have been proposed to explain IPO underpricing and long-run underperformance. In addition, both anomalies are studied form the viewpoints of two competing finance theories: efficient market hypothesis and behavioral finance. Theories of underpricing are grouped within two broad categories: asymmetric information based models and behavioral theories. While asymmetric information based models assume that one of the IPO transaction parties knows more than others, and that these information frictions give rise to underpricing, behavioral explanations, on the other hand, assume the presence of irrational investors who are the prime cause of underpricing. Theories of poor long-term performance are based on behavioral finance perspective only, where "investor sentiment" plays the main role. On the contrary, proponents of market efficiency strongly argue that the notion of systematic IPOs long-run underperformance is spurious. Secondly, the thesis empirically examines the presence of underpricing and the long-performance of IPOs in European NYSE Euronext markets. In general, the results undoubtedly show that IPOs in the sample are moderately underpriced on average. However, the assessment of IPOs long-run performance provides contentious findings and probably requires further research.
|
|
Methodology of transgenic mouse preparation.
Reslová, Gabriela ; Komrsková, Kateřina (advisor) ; Pěknicová, Jana (referee)
The thesis deals with methods used for preparation of transgenic mice and with comparison of them in terms of efficiency and suitability of applications for research purposes. Technologies for the transfer of gene constructs involve microinjection of DNA into the pronukleus of fertilized oocyte, which seems to be the most reliable one. Another possibility is associated with the use of sperm, which carry the incorporated gene construct into the oocyte during in vitro fertilization, or with the use of retroviral vectors by which the oocytes are transfected. Somatic cell nuclear transfer into enucleated oocyte, the use of embryonic stem cells which are incorporated into strange embryo or transgenesis by liposomes are techniques far less explored and not so often used. The next section of this thesis introduces to the reader individual steps which are necessery for successful transgenesis. It is focused on ovarian stimulation which is necessery to obtain sufficient amount of oocytes, as well as on stimulation of recipient mother in embryo transfer as well as on capacitation of sperm required for in vitro fertilization. The thesis also deals with backward analysis confirming positivity of transgene expression by genotyping of pups and by crossbreeding of transgene positive and compares advantages and...
|
|
Plant alkaloids and their effects on enzymes metabolizing xenobiotics
Višněvská, Kateřina ; Stiborová, Marie (advisor) ; Černá, Věra (referee)
Sanguinarine and chelerythrine are quaternary benzo[c]phenanthridine alkaloids. The first step in sanguinarine metabolism is its reduction to dihydrosanguinarin. Antimicrobial and anti-inflammatory activities of these alkaloids are used in dentistry and as feed additives. Sanguinarine and chelerythrine induce apoptosis of cells. Fluorescence of these alkaloids and intercalation into DNA could be utilized to use the alkaloids as supravital DNA probe. Negative effect of sanguinarine and chelerythrine is their genotoxicity. Cytochrome P450 and peroxidase oxidize ellipticine to detoxication and activation metabolites. Ellipticine is a potent antineoplastic agent exhibiting the multimodal mechanism of its action. Ellipticine intercalates into DNA and inhibits topoisomerase II. Covalent DNA aducts are mediated by CYP or peroxidase oxidation of ellipticine. The anti-tumor activity of ellipticine and its derivatives is caused by a combination mechanism of cell cycle arrest and induction of the apoptotic pathway. Pharmacological efficiencies and geneotoxic side effects of ellipticine is dependent on levels and activities of cytochrome P450 or peroxidase in target tissues. Aristolactams are the major metabolites of biotransformation of aristolochic acid. Nitroreduction is the crucial step in formation of an...
|
|
Development of transient transfection protocol for HEK293 EBNA1 cells
Šmíd, Jiří ; Španová, Alena (referee) ; Ševčík, Mojmír (advisor)
Recombinant proteins belong to considerable biofarmaceutics products used in biomedical research and in the treatment of human disease. Recombinant protines can be produced by stable transfection in big amount or by faster transient transfection with smaller amounts. To provide regular biological activity, it is necessary for the protein to be properly folded and post-translationally modified. As these modifications can be accurately performed only in mammalian cells, they have become the major host for complex r-protein expression. In this thesis is described transient transfection HEK 293 EBNA1 cells with linear polyethylenimines. These cells has been adapted to suspension cultivation in serum free medium. The cells were transfected with pcDNA3.1, pCI, pEBSV1, pCEP4, pEAK8 a pcDNA5/FRT/TO plasmids, everyone contained repoter gene SEAP. Concentration of SEAP in cell culture supernatants were determined in order to compare efficiencies of individual transfections. DNA:PEI ratio was another factor which was optimised and two different PEIs were compared. Highest achieved expresion was 50 mg per litre with transfection in 24 well plate when DNA:PEI ratio was 1:5. Comparison of six different plasmids give the bigest expresion pCEP4/SEAP, in well plate as well as in scaled up system.
|
|
Enhancement of the efficacy of DNA vaccines against the cervical cancer with helper epitopes
Peřinová, Lucie ; Brábek, Jan (referee) ; Šmahel, Michal (advisor)
The human papillomaviruses (HPV) are the etiological agent of cervical cancer. Their oncoproteins E6 and E7 are involved in the transformation of an infected cell into a neoplastic cell, thereby they are the target antigenes for the development of DNA vaccines. Helper epitopes activating CD4+ T cells are under study because they enhance the efficacy of DNA vaccines through increasing the number of cytotoxic T lymphocytes and thereby removal of the tumor. There are already being used epitopes derived directly from oncoproteins, synthetic epitopes or bacterial epitopes for the general enhancement of the immune response. Sufficient number of comparative studies which would establish the exactly most efficient helper epitopes has not been made. The research aims at combining more peptide types using immunostimulatory molecules.
|
| |
guest ::
