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Vliv odvodnění a vegetace na množství a diverzitu metanogenů v rašeliništích
LÍSALOVÁ, Olga
This work deals with attribute of microbial communities, especially methanogenic Archaea in pristine and drained peatlands. Methanogens are strictly anaerobic microorganism thus they live in deeper anoxic layers of peatlands. Therefore, the aim of this study was to determine how the drainage affects the amount and diversity of methanogens depending on different depths, types of peatlands and litter type.
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CHR-test-method of excercise testing by endurance sportswomen related to renal functions
Šturmová, Anita ; Radvanský, Jiří (advisor) ; Procházka, Michal (referee)
The aim of this study was to familiarize and imlement methodics of CHR-test. For using exercise testing were used endurace trained women. The results of exercise testing were compared with previously published study for untrained probands. The paper discussed resulting indexes of testing depending on individual fitness and the conslusiveness itselfs of this type of testing, that is not expanded to date. The next part of the thesis presents the results of experimental work: relations between stress and reaction of kidney in the form of microalbumiuria and transferinuria. Powered by TCPDF (www.tcpdf.org)
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Characterization of collagen from different animal tissues
Mikulíková, Zuzana ; Drábková, Michaela (referee) ; Márová, Ivana (advisor)
The aim of the proposed diploma thesis is to optimize the procedure of collagen isolation from animal tissues and methods for molecular characterization of isolates. In experimental part, isolations of collagens from animal tissues (chicken, hen, carp, and pork skin) were performed. Total protein concentration was determined by Biuret method and by TNBSA method. Both methods were used also in order to study physiological and thermal stability of the isolates under model conditions. Isolated collagens were characterized at molecular level using PAGE–SDS and microfluidic electrophoresis. Further, amino acids composition and microelement content were analyzed. Finally, the stability of isolated collagens in several types of simulated physiological and pathological conditions was tested too. During isolation relatively sufficient collagen yields were obtained (about 15 % of purified collagen per original mass of biological material). In all analyzed tissues Type I collagen was found. Thermal stability of individual samples differed according to biological source type and tissue age. Viscosimetry measurements confirmed higher stability in collagen samples of older animals. Collagenase exhibited the lowest degradation effect to bovine collagen, while selected mixed microbial hydrolases differed according to enzyme preparative type. Incubation of collagen isolates with selected human pathogens confirmed higher resistance of bovine collagen to biofilm formation when compared with the chicken one.
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Study of metabolic changes in carotenogenic yeasts cultivated under different conditions
Starečková, Terezie ; Breierová, Emília (referee) ; Márová, Ivana (advisor)
The aim of this diploma thesis realized as a comparative study was the study of regulation of carotenoid and ergosterol production in several carotenogenic yeast strains. Yeasts were exposed to exogenous stress factors. Salt stress and oxidative stress (hydrogen peroxide) were reached by addition of NaCl and hydrogen peroxide into production media. Complex changes on metabolome (e.g. pigment and ergosterol production, RP-HPLC), proteome and genome were followed. Proteome changes were analyzed by PAGE-SDS and 2D electrophoresis. To isolation and analysis of chromosome DNA pulsed field gel electrophoresis (PFGE) was used. Six yeast strains were enrolled into the comparative study; three strains of the genus Rhodotorula and three strains of the genus Sporobolomyces. While yeasts Rhodotorula sp. were characterized by enhanced biomass as well as carotenoid production in normal and stress conditions, production of biomass by Sporobolomyces sp. was substantially lower. Carotenoid production in Sporobolomyces sp. was higher than in Rhodotorula sp.; the highest increase of was beta-carotene production was observed in Sporobolomyces salmonicolor cells stressed by salt (4x higher than in control) or peroxide (5x higher). Proteins were isolated from yeast cells by combination of mechanical and chemical disruption by glass beads and NaOh or SDS. Better yields were obtained by NaOH. Two staining methods were tested in PAGE-SDS protein analysis. Coomassie Brilliant Blue staining exhibited lower sensitivity, silver staining led to better visualisation of minor protein fractions too. 1D protein profiles was difficult to evaluate, therefore, 2D electrophoresis of selected strains (R.glutinis, R.rubra) was done. In yeast genome analysis by PFGE at minimum 7 DNA fractions were observed. These results probably are not final, further study will be needed for detailed characterization of red yeast genome.
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Inovace klasifikace a zkoušení podložních zemin pro realizaci lesních odvozních cest
Tvrdý, Daniel
Basic presumption of good functionality of forest roads is not only high quality of laminars in flexible pavement, but also right distribution of elasticity modules in soil plain and it's protection against degradation during the process of building. Materials used for TA ČR project, which were isolated from forest roads subsoil, were analyzed in soil mechanics laboratory of Geostar s.r.o. company for the purposes of diploma thesis. Basic laboratory tests were carried out in order to classify soils. Sub-component is to propose a simplified classification of soils for the implementation of forest haul routes. In this work are validated according to the static load tests ( SZZ ) and dynamic light boards ( LDD ) . The thesis includes evaluation of the quality of existing forest roads for faults and failure on forest roads classes 1L and 2L with different surfaces and ground covers .
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The mechanism of action of anticancer drug ellipticin in target tissues of its effect
Vranová, Iveta ; Stiborová, Marie (advisor) ; Martínková, Markéta (referee)
Ellipticine is an alkaloid isolated from Apocynaceae plants exhibiting significant antitumor and anti-HIV activities. Cytochromes P450 (CYP) and peroxidases are the enzymes participating in metabolism of ellipticine. This process provides activation and detoxication metabolites of ellipticine. The CYP enzymes, which participate in oxidation of ellipticine in different tissues (liver, lung and kidney) of rat, a model organism simulating the fate of ellipticine in humans have already been identified. In this work, the effects of ellipticine on contents and catalytic activities of CYPs and other components of the mixed-function oxidase (MFO) system in this animal system were studied. For detection of contents of CYPs and other components of the MFO system, spectroscopic and electrochemical methods were used. To determine catalytic activities of CYPs and NADPH:cytochrome P450 reductase, reactions with specific substrates of these enzymes were utilized. The results found in this study demonstrate that expression and catalytic activity of CYP1A is induced by ellipticine in all of the tested organs (liver, kidney and lung) of rats treated with the drug. Moreover in liver, the cytochrome b5 expression is also induced. In addition, in this organ, expression and catalytic activity of CYP3A was increased by...
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Supercritical Fluid Extraction and Insecticidal Activity of Lamiaceae Plants
Sajfrtová, Marie ; Pavela, R. ; Karban, Jindřich ; Sovová, Helena
Natural insecticides based on bioactive extracts from plants represent a new trend in plant protection. Supercritical extraction (SFE) using CO2 is a modern extraction technique able to isolate high-added value compounds from plant without any thermal degradation, which allows to obtain extracts with high biological activity. In this work, insecticidal activity of SFE extracts from plant of mint family (Lamiaceae) has been studied and compared to activity of isolates obtained by conventional extraction techniques (hydrodistillation, organic solvent extraction). As a result of variation in extraction pressure, three types of extract were obtained: extract rich in volatile compounds (at 12 MPa), extract rich in oleoresins (at 28 MPa) and in polar compounds (5% of acetone used as modifier). Chemical composition and biological activity (LD50, antifeedant activity) of all isolates were evaluated and compared.
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Isolation and purification of the environmental DNA from the horizon of the capping: stratification study
Zahradník, Jiří ; Šulc, Miroslav (advisor) ; Weignerová, Lenka (referee)
According to our contemporary knowledge soil is the most abundant source of microbial biomass. Unfortunately, only one percent of the microorganism species is available by classical cultivation techniques. Soil metagenomic DNA is a collection of the whole DNA including also uncultivated microorganism in the soil sample and provides information to study molecular aspects of microorganism and their DNA sequences inaccessible by other techniques. This work is focused on characterization and isolation of soil metagenomic DNA from deep horizon of capping. Evaluation in term of the isolation and techniques of recombination of DNA and stratification study are included in this work. Obtained collection of samples was preliminary characterized with the view of quality - content of clay, humic compounds and crude number of microorganism. The purified soil metagenomic DNA was quantitatively and qualitatively characterized for each sample. The quantification method and DNA quality determined the next applications and procedures of DNA techniques. Also the soil quality was discussed from this point of view. According to the results of DNA analysis, the three selected DNA samples were processed to DNA library with 16S rRNA DNA loci and after DNA sequencing analysis the phylogenetic study was performed. This...
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The occurrence of the HW element in Welsh English
Kolísková, Barbora ; Skarnitzl, Radek (advisor) ; Šturm, Pavel (referee)
The aim of this diploma thesis is to investigate the accent realizations of the initial consonant in wh- words in Welsh English. The theoretical background of this study deals with the development of Welsh language and its interactions with the British standard variety. Together with the previously mentioned the phonological inventories of Welsh, RP, and Welsh English are described in the first part. We further analyzed the development of the <hw> element in the British Isles in general. The research part of this study is concerned with the analysis of the data from 20 speakers of Welsh English. Three speech styles were used for the final analysis: spontaneous speech (informal style), isolated words (formal style) and reading of Cinderella (formal style). Signal-to-noise ratio and voicing of the <hw> segments were measured in order to discover the occurrence and the nature of the <hw> element. The lowest levels of harmonicity were measured in segments that were obtained from the formal reading style. The highest levels were measures in the spontaneous speech tokens. Younger speakers showed higher levels of harmonicity than older speakers. The results of the male and the female participants were different in each type. In the spontaneous speech style the male speakers tended to have lower levels of...
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Important roles of heme as a signal and a gas-sensing site: heme-sensing and gas-sensing proteins
Fojtíková, Veronika ; Martínková, Markéta (advisor) ; Hudeček, Jiří (referee)
Heme-containing sensor proteins are heme proteins, which are divided into two groups: heme-sensing and gas-sensing proteins. The function of heme-sensing proteins is affected by heme availability. Association (or dissociation) of heme moiety of heme-sensing protein regulates various physiological functions, including protein kinase activity, transcription and other important functions essential for cell survival. In gas-sensing proteins, heme acts as the sensing site for binding of gaseous molecules (including O2, NO and CO) and indirectly regulates physiological functions, including protein kinase activity, transcription and other important functions essential for cell survival. The recent studies on heme-containing senzor proteins published in scientific journals are summarized in this thesis. The experimental part of this thesis focused on the specific heme-containing sensor protein - a globin-coupled histidine kinase from Anaeromyxobacter sp. strain Fw 109-5 (AfGcHK). The aim of this thesis was to amplified and isolate plasmid carrying gen for AfGcHK. Consequently the protein was expressed in E.coli BL-21(DE3) and the protein was isolated. Based on the results, the isolation process was optimized. Moreover, the purified preparation of isolated AfGcHK was prepared in more than 99% of homogeneity....
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