|
|
Fluorescence Spectroscopy: Advanced methods and their defined applications in protein science
Pospíšil, Petr ; Hof, Martin (advisor) ; Heřman, Petr (referee) ; Polívka, Tomáš (referee)
The hydration and dynamics of the biomolecules appear to be vital for their proper biological functioning. In the presented thesis, various fluorescence techniques were developed and applied to access these properties and their changes upon the mutual interactions of the biomolecules. Initially, the solvent relaxation method based on recording time-dependent fluorescence shift (TDFS) was used to map DNA interactions with proteins and lipids by the newly synthesised fluorene dye covalently bound to the DNA. Secondly, copper-transporting ATPase was probed by Badan attached to the copper-binding cysteine-proline-cysteine motif. The variations in hydration were found to be crucial for the proper ATPase function. Third, a detailed study on quenching of Badan/Prodan fluorescence by tryptophan revealed the limitations of the TDFS method for protein studies, which is essential finding for further applications of TDFS. Fourth application involves investigations of heavy atom effects on the excited state relaxation processes by up-conversion approach in iodinated metallocorroles, which are promising dyes for biological imaging. The obtained findings shall help in further tuning of the optical properties of the corroles desired for the variety of applications. Finally, fluorescence correlation spectroscopy...
|
|
| |
|
|
The History of the Sokol Movement in Lysé nad Labem
Herman, Petr ; Kašpar, Ladislav (advisor) ; Zachariášová, Jana (referee)
Sokolská organizace má za sebou bohatou minulost a je navždy zapsána do historie tohoto státu. Z jejích řad vyšlo mnoho obránců naší republiky, ať už jako legionáři nebo bojovníci za svobodu na bojištích druhé světové války. Sokol z nich vychoval lidi s vynikající morálkou i fyzickou kondicí. Lyský Sokol vznikl jen pár let po "průkopnickém" Sokole v Praze a hned byl velký zájem o členství v jednotě. Bohužel tragickou nehodou ve městě, která vyvolala krizi, byla činnost jednoty záhy na 2 desetiletí neúmyslně pozastavena. Naštěstí obyvatelé nezapomněli a Sokol znovu ožil a dále sílil. Tentokrát organizaci do cesty vstoupila válka, a tak činnost ochabla. Po válce se vzchopil a zažíval svá nej lepší léta. S druhou světovou válkou byla činnost Sokola opět zastavena a tentokrát ani po skončení neměl příliš prostoru k rozletu, komunistická diktatura ho též shledala nepřípustným. Přesto se Sokolové nevzdali a přes mnoho nepravostí působil v ilegalitě. Revoluční rok 1989 mu opět otevřel brány. Tak jak byl Sokol v rozkvětu za první republiky, si však dnes můžeme nechat zdát, ačkoliv dle mého názoru jsou jeho myšlenky stále aktuální. Bohužel dnešní společnost, jak se zdá, upřednostňuje zřejmě jiné cíle a hodnoty.
|
|
|
Deconvolution fluorescence microscopy of yeast cells
Štec, Tomáš ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Deconvolution fluorescence microscopy of yeast cells Author: Tomáš Štec Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jarmoír Plášek, CSc., Institute of Physics of Charles Uni- versity Abstract: Fluorescence microscopy presents an fast and cheap alternative to more advanced imaging methods like confocal and electron microscopy, even though it is subject to heavy image distortion. It is possible to recover most of the original distortion-free image using deconvolution in computer image processing. This al- lows reconstruction of 3D structure of studied objects. Deconvolution procedure of NIS Elements AR program undergoes an thorough inspection in this diploma the- sis. It is then applied on restoration of 3D structure of calcofluor stained cell wall of budding yeast Saccharomyces cerevisiae. Changes of the structure of the cell wall during cell ageing are being examined. Cell wall of aged cells shows increased surface roughness and even ruptures at the end of cell life. Keywords: fluorescence, microscopy, deconvolution, NIS Elements AR, calcofluor, yeast, cell wall, ageing
|
|
| |
|
|
Study of the relations between the structure and function of C - terminal vanilloid receptor TRPV1
Gryčová, Lenka ; Obšil, Tomáš (advisor) ; Heřman, Petr (referee) ; Urbánková, Eva (referee) ; Krůšek, Jan (referee)
Transient receptor potential channel vanilloid receptor subunit 1 (TRPV1) is a thermosensitive cation channel activated by noxious heat as well as a wide range of chemical stimuli. Although ATP by itself does not directly activate TRPV1, it was shown that intracellular ATP increases its activity by directly interacting with the Walker A motif residing on the C-terminus of TRPV1. In order to identify the amino acid residues that are essential for the binding of ATP to the TRPV1 channel, we performed the following point mutations of the Walker A motif: P732A, D733A, G734A, K735A, D736A, and D737A. Employing bulk fluorescence measurements, namely a TNP-ATP competition assay and FITC labeling and quenching experiments, we identified the key role of the K735 residue in the binding of the nucleotide. Experimental data was interpreted according to our molecular modelling simulations. Calmodulin (CaM) is known to play an important role in the regulation of TRP channels activity. Although it has been reported that CaM binds to the Cterminus of TRPV1 (TRPV1-CT), no classic CaM-binding motif was found in this region. In this work, we explored this unusual TRPV1 CaM-binding motif in detail and found that five residues from a putative CaM-binding motif are important for TRPV1-CT's binding to CaM, with arginine R785...
|
|
|
Lepidoptera Community of the Woodland Complex Loučany near Klatovy
Rendlová, Veronika ; Heřman, Petr (advisor) ; Hanel, Lubomír (referee)
This thesis is focused on faunistic survey of Macrolepidoptera in Loučany (foothills of the Šumava Mts, Czech Republic). The research, involving different capture methods, proceeded from April to November 2015 on given transect and several other habitats. Based on the results (including partial data from 2013 and the last two decades of the 20th century), an inventory of identified species of Macrolepidoptera was made. Altogether 205 species from 16 families were found in Loučany. The most important species are for example the Black-veined White (Aporia crataegi), High Brown Fritillary (Argynnis adippe), Pearl-bordered Fritillary (Boloria euphrosyne), Purple Emperor (Apatura iris), Spruce Carpet (Thera britannica) and Sandy Carpet (Perizoma affinitata). The environmental parameters of experimental sites were noticed during research and the habitat preferences of species were analysed using specialized literature. Altogether 186 species (90,7 % of all identified species) are linked to some kind of woodland habitat. To support present biodiversity of the community of Macrolepidoptera, it is necessary to practice a suitable woodland management, conserving mosaicity and open canopy. Key words: Lepidoptera, species diversity, Czech Republic, Klatovy area, conservation, woodland management
|
|
| |
|
|
Response of plasma membrane potential and intracellular pH of yeast to changes in the concentration of extracellular K+
Babuka, David ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
The bachelor thesis is focused on studying yeast cells and their response to various external conditions. Main focus was on the study of intracellular pH and membrane potential change under the condition of varying extracellular concentration of K+ ions. In particular we studied to what extent are the yeast cells able to compensate these changes. The ability of yeasts to resist the changes of external pH of the cell medium was studied in an experiment complementary to the measurements of intracellular pH. To measure the intracellular pH a genetically encoded fluorescent probe ratiometric pHluorin was used and to measure the changes of membrane potential a fluorescent probe diS-C3(3) was used. Also we successfully applied a method of synchronously scanned fluorescence to supress the cell autofluorescence.
|
|
|
Proposal for Using Virtual Reality Technology in Marketing of a Company
Šmerda, David ; Herman, Pavel (referee) ; Kaňovská, Lucie (advisor)
This Bachelor Thesis describes the current advent of virtual reality (VR) technology and possible use of this technology in marketing of the company. I will apply this technology to PTÁČEK - velkoobchod a. s. In the first part I will focus on the theory of marketing, marketing communication, marketing trends including virtual reality and psychological aspects in connection with VR. In the second part I will introduce the company PTÁČEK - velkoobchod a. s. And using selected analyzes I will analyze its marketing environment, which I will summarize in the resulting SWOT analysis. The third part contains a proposal to improve the company's marketing through VR technology, it‘s implementation and solution in practice.
|
guest ::
RSS feed.