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Study of retention behaviour of neopterine in HILIC chromatographic system with different stationary phases
Matula, Aleš ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
TITLE: The study of neopterin retention characteristics in HILIC chromatography system with different types of stacionary phases SUMMARY: This graduation thesis was dealing with the development of an analytical method for the identification and quantification of biologically active substances biopterin, neopterin and 7,8-dihydroneopterin by hydrophilic liquid interaction chromatography (HILIC) connected to fluorescence detection. Neopterin represents an important and useful parameter for monitoring of the immune system activation, that accompanies different infectious and autoimmune diseases. Increased level of neopterin concentration can be observed in several types of malignant tumors. Recently the coherency between neopterin, 7,8-dihydroneopterin and intracelular oxidative stress and apoptosis of the cells has been demonstrated. Totally reduced form of biopterin serves as a cofactor of some enzymes in human organism. HILIC is an option of high performance liquid chromatography (HPLC). Three chromatographic columns (LUNA HILIC, OBELISK R, OBELISK N) were tested at different chromatografic conditions. Mobile phase consisted of mixture of aqueous component (ammonium acetate buffer, acetic acid, water) and water miscible organic component (acetonitrile). The mixture of acetonitrile and 100mM ammonium acetate...
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Thermal analysis of interaction of epidermal lipids IV
Komárková, Radka ; Musilová, Marie (advisor) ; Doležal, Pavel (referee)
Thermal analysis of interaction of epidermal lipids IV Radka Komárková, diploma paper, June 2009 Abstract Main function of skin is protect organism in face of chemical and mechanical damage. Skin is composed of several layers, and most considerable barrier of which forges most upper part stratum corneum. Stratum corneum contains cells fill in with keratin, which are surrounded lipid matrix. Main components of lipid matrix are ceramides (50%), cholesterol (25%) and fatty acids (10%). At present are many experimental works specialized up to preperation in vitro lipid membrane and aim of our work is find out, whether concentration of water can influence hydration lipophilic components of lipid membrane. We used samples, which had constant composition lipid mixture (cholesterol : ceramid 14S24, 1:5) and they differentiated content of water (75%, 50% and 25%). This samples were remit to several cycles of cool down and warming up by the help of DSC, where was establishment only one mixed peak of lipid mixture. All samples showed out interaction with water, however sample with 25% substantiality, it was rather on limits of analytic mistake.
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Sport activities in human-dog interaction
Grbavčic, David ; Gnad, Tomáš (referee) ; Neuman, Jan (advisor)
Thesis title Sport activities in human - dog interaction Thesis objectives The thesis introduces sport activities involving the interaction of human and dog. Its aim is to ascertain the physiological load imposed on the body during selected sport activities in which a human is engaged in an interaction with a dog and to compare the findings with the physiological load generated by another sport activity (mountain bike). Methods Analysis of expert documents. Heart-rate monitoring with the Polar RS400 sporttester capable of processing data on heart-rate and body load. Results The results obtained in separate measurements have proven the defence work to be more physically demanding for a sportsperson than riding a mountain bike. The assumption was confirmed by the differences in heart-rate values and the varying time periods needed for the sportsperson's recovery. Recovery time after a mountain bike training session is fourteen hours. In contrast, it takes the sportsperson (the helper) twenty-seven hours to recover from a defence work session of equal length. Considering the diversity of defence work training units and the small number of measurements the results are to be taken as tentative. However, they will facilitate improved training preparation of defence work helpers. Key terms dog sport,...
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Teacher`s influence on social climate in elementary school
Innemanová, Jana ; Valentová, Lidmila (referee) ; Hadj - Moussová, Zuzana (advisor)
The diploma work describes a research done in the social climate of the observed class and its development in the course of a school half-year. The main task was to discover the social climate in the class and the possibilities of its affecting through a pedagogue's influence. The work finds out how the social climate is affected by mutual interaction between the teacher and his/her pupils and how a positive climate of the class affects the pupils' prosperity in the learning process.
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Protein-protein interaction mapping of cytochrome P-450 by methods using chemical modification and mass spectrometry
Ječmen, Tomáš ; Černá, Věra (referee) ; Šulc, Miroslav (advisor)
Cytochromes P-450 (P450s) belong to haemoprotein superfamily and they are responsible for metabolism of a wide variety of compounds, among others many drugs and carcinogens. P450s serve as the terminal oxidases in the mixed function oxidase system in cooperation with a redox partner NADPH: cytochrome P450 reductase (CPR) providing input of two electrons to the reaction cycle of P450. The CPR can be substituted by other redox partner of P450, cytochrome b5 (cyt b5), to deliver the second electron. Three dimensional structure of P450 is required in order to fully understand its reaction mechanism. At the present time, a homology model of cytochrome P-450 2B4 (CYP 2B4) is available in our laboratory. In this study, the mapping of interaction domain between CYP 2B4 and cyt b5 employing a crosslinking agent EDC to form amide bonds between close complementary charged amino acid side chains was the first goal. We have identified five interacting amino acid pairs in total using mass spectrometry (MS). The second research interest was to verify and refine the CYP 2B4 model using a photoaffinity labelling with N-(p-azidobenzyl)-N-methyl-p-aminophenylamine probe. This photoreactive probe is known as CYP 2B4 ligand binding to the central iron atom of haem. After photoactivation the arginine 197 was found by MS...
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Diversity of methods used for characterization of molluscan hemocytes
Jindrová, Zuzana ; Skála, Vladimír (referee) ; Horák, Petr (advisor)
Hemocytes are the main immune cells of invertebrates; therefore they can be found in molluscs, too. They differ both in morphology and function. The two generally accep- ted morphological types, granulocytes and hyalinocytes, vary in the level of phagocy- tosis and encapsulation, production of reactive oxygen species and nitrogen oxide, and presence of some enzymes. There is an array of methods by means of which hemocy- tes can be characterized. Microscopy serves particularly for study of morphology. An- tigens localized on the surface can be determined by monoclonal antibodies or lectin probes. Hemocytes can be divided on the basis of cell size and granularity using gra- dient centrifugation or flow cytometry. Production of nitrogen oxide and reactive oxy- gen species is monitored by adding appropriate substrate which changes its proper- ties after reaction with the radical. It may become fluorescent, change absorbance of the solution or form a visible precipitate. Another possibility is the use of chemilu- miniscence. The objective of hemocyte research is to explain mollusc-pathogen inter- action. 1
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Error in Descartes' philosophy
Klinka, Tomáš ; Hill, James (referee) ; Palkoska, Jan (advisor)
The present work analyses the theory of error of René Descartes, mainly in the context of his Fourth Meditation. This analyze begins with the exposition of the Descartes' theory of judgment and his notions of understanding and volition. The main question is the following: Is Descartes' theory of error valid, even if we consider all the objections of Descartes' critics? The maill objection is: in the explication of error during our judgment as a result of interaction between a limited understanding and an unlimited human will, we have to consider us capable do decide at will what we believe and what we don't, which is not an intuitive position. But this work suggests that this non-intuitive position is sustainable, if we abandon the "strong" version of Descartes' voluntarism, in which the will influences our believes directly, and we see Descartes' theory of error as using the "milder" version of voluntarism (called here "attention" voluntarism), in which the will influence us indirectly, by forcing our attention in the needed way. As a final conclusion, we consider Descartes' theory of error as valid.
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Possibilities of pre-travel consultancy and specific prophylaxis of imported infectious diseases in the region of South Bohemia
RYTÍŘOVÁ, Zuzana
The theoretical part of the thesis deals with the problems of imported infectious diseases. It comprises the factors influencing their incidence and the possibilities of specific and nonspecific prophylaxis and prevention. Further on, this part focuses on chosen infectious diseases, their epidemiological and clinical characteristics and trends in incidence. These infections are divided according to possibilities of specific prophylaxis. The thesis offers an overview of the possibilities of pre-travel consultancy in the South Bohemia region. The practical part of the thesis aims at the characterization of the incidence of imported infectious diseases in the districts within the region of South Bohemia in the years 2000 ? 2010. It also presents a survey of medical facilities and other subjects providing pre-travel consultancy and specific prophylaxis in the sphere of imported infectious diseases prevention. During the monitoring period, the total number of imported diseases which occurred in the region of South Bohemia and were registered in the EPIDAT system was 351. Out of this number, 205 cases occurred in the district of České Budějovice. (Český Krumlov ? 47, Jindřichův Hradec ? 25, Písek ? 2, Prachatice ? 17, Strakonice ? 31, Tábor ? 24). The most frequent imported infections were alimentary infections (240 cases, i.e.68 %); out of this, the most frequent cases were diseases caused by Salmonella (73 cases), Shigella (71 cases) and campylobacters (42 cases). The most frequent countries of origin were Egypt (43 cases), Croatia (42 cases) and India (30 cases). The number of medical facilities and other subjects providing pre-travelling consultancy and specific prophylaxis of infectious diseases rose from nine to thirteen during the period of monitoring. Four hypotheses were determined within this thesis. Hypothesis no.1 ? during the period of monitoring the total number of imported infectious diseases increased. Hypothesis no.2 ? during the period of monitoring the total number of infectious diseases imported from exotic destinations increased. Hypothesis no. 3 ? during the period of monitoring there was an increase in the number of the facilities and other subjects providing pre-travel advice, vaccination or other specific prophylaxis. Hypothesis no. 4 ? the increasing number of medical facilities or other subjects providing pre-travel advice, vaccination or other specific prophylaxis did not influence an increase in the number of imported infectious diseases during the period of monitoring. Only hypotheses no. 3 and no. 4 were proved.
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The influence of repeated defrosting on nutritional efficiency of Artemia salina nauplii for early developmental stages of fish
HULAN, Petr
Nutritional efficiency of deeply frosted and repeatedly defrosted zooplankton has not been totally clarified, and it is often questioned, however, the nutrients composition and effect of such deteriorated feed are missing. The aim of this study was to assess nutritional eddiciency of repeatedly defrosted nauplia Artemis salina, as well as changes in their chemical composition on the basis of the growth, survival and presence of the deformations in early ontogeny of Japanese medaka ( Oryzias latipes). The samples of fresh hatched artemia nauplii served as a control group. The experiment was carried out with samples of live nauplii, permanently frosted at -14°C, defrosted to 4°C and again frosted and defrosted to 25°C and frosted again. The larvae of medaka in the period of one week after shift to the exogenous nutrition were examined. Before the commencement of the experiment the composition of samples in terms of nutrient, amino acids and fatty acid contents was determined. The results proved that Oryzias latipes fed with live artemia not only grew better but also survived more that other fis in other treatments. No deformations were recorded Fish fed frozen food survived better than those fed with live Artemia. It was also found that fish fed live Artemia seems to be the highest however from the amino acid point of view it seems to be more appropriate to freeze it to -14°C, defreeze to 25°C and refreezing to -4°C. as for the fatty acids Artemia adjusted by the permanently freezing to -14°C. The processes of thawing and repeated freezing of artemia nauplii result in leaching of some nutrients but in the thickening of some others.
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Studies on NKR-P1A and NKR-P1B receptors expressed in eukaryotic organisms
Ivanova, Lyubina ; Rychlovský, Petr (referee) ; Bezouška, Karel (advisor)
NK (natural killer) cells, with their ability to identify antigens and extraneous substances, available in the organism through various moleculary receptors, are an important component of the immune system. The NKR-P1A and NKR-P1B proteins belong to the lectin receptors of natural killer cells. Primary ligands of lectin receptors comprise terminal oligosaccharides of glycoproteins on the surface of target (e.g. tumor) cells. The interaction between carbohydrate structures on the surface of antigens and their binding partners on NK receptors is followed by triggering the effector function of NK cells against the targets. The NK cells and NK receptors findings and their interactions with ligands are greatly utilized in the treatment of cancer, viral and autoimmune diseases. Heterologous protein production in the eukaryotic organism brings a lot of advantages. Unlike the prokaryotic organism, the methylotrophic yeast Pichia pastoris has the capability of performing many posttranslational modifications resulting in production of biological active protein molecule. Usually, the P. pastoris expression system disposes of high level protein expression and is also generally regarded as being faster, easier, and less expensive to use than expression systems derived from other eukaryotes. In this thesis, I...
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