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Study of factors influencing proton transport mediated by MntH protein.
Straková, Eva ; Urbánková, Eva (advisor) ; Žáčková, Markéta (referee)
The natural resistance-associated macrophage proteins (Nramp) form a family of secondary active transporters facilitating the transport of divalent metal ions (Mn2+, Fe2+, Cd2+, Co2+, Zn2+) across biomembranes. Bacterial proteins belonging to the Nramp family were describedas proton-dependent manganese transporters - MntH. MntH homolog from E. coli representing a model system for structure function relationship was used in this study. Metalinduced proton transport mediated by MntH was measured by means of changes in intracellular proton concentrations using pH sensitive fluorescent protein pHluorin. This approach does not allow proper estimation of real quantity of transported protons, since the internal pH is tightly regulated. The cellular buffering capacity was calculated from measured pH changes induced by addition of propanoic acid, in order to better quantify the transport. The changes in [H+] induced by cadmium were recalculated using buffering capacity for the wild type MntH and its single-point mutation N401G, known to influence intracellular pH. It was shown that the chosen approach significantly influences results, including the dependence of transport on external pH. Potential influence of cadmium to respiration was imitated by using inhibitors of respiratory chain (KCN, NaN3) in measurements...
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Ionizing radiation shielding simulation using MCNP code
Konček, Róbert ; Košťál,, Michal (referee) ; Katovský, Karel (advisor)
Radiation is defined as ionizing if it has enough energy to remove electrons from atoms or molecules when it passes through or collides with matter. This ability implies potentially detrimental effects on living tissue. Ionizing radiation shielding is therefore a discipline of great practical importance. The thesis builds upon the author's previous work on the topic and widens the scope of discussion with theoretical and practical issues of advanced shielding calculations. The theoretical part of the thesis describes several approaches to calculating fluence or absorbed dose at an arbitrary point in space. Point-kernel methods provide sufficiently accurate results for simpler shielding problems. In many practical cases, however, calculations based on the transport theory are necessary. There are two basic types of transport calculations: deterministic transport calculations in which the linear Boltzmann equation is solved numerically, and Monte Carlo calculations in which a simulation is made of how particles migrate stochastically through the problem geometry. Advantages and disadvantages of both methods are discussed. In the practical part are the results of radiation shielding calculations performed with a major Monte Carlo code - MCNP6, compared with those obtained in the experiments, which were carried out at the Ionizing Radiation Laboratory at Department of Electrical Power Engeneering, FEEC BUT. The experiments consisted of placing a cobalt-60 radioisotope source at three different positions inside a lead collimator, and counting pulses with two different scintillation detectors positioned in front of the opening of the collimator, alternately with or without lead shield located between the source and the used detector. Agreement of the calculations and the data from the measurements is reasonable, given the inherent uncertainties of the experimental set-up. Performed sensitivity analysis shows relative importances of different parameters used as inputs in simulations, such as densities of materials, or dimensions of the scintillation crystals. Annotated MCNP input files used for simulation are also part of the thesis.
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Signal analysis of transport and stochastic processes of emission detectors
Míšek, David ; Tofel, Pavel (referee) ; Andreev, Alexey (advisor)
This Diploma thesis deals with properties of CdTe detectors. This material is ranked among optical sensitive group. This thesis can be thematically divided into two basic parts. The first part describes properties of both elements and the chemical adduct. Selected properties show perfections of the material, which are specializing it to optical area of application. The end of this part contains description of contact metal-semiconductor. This problem is important to comprehension principle the material contact. In second part there were making many measurements. For more accurately compare were first measurements doing without light and then with light. During making measurement was changed wave length and temperature of sample. Key factors were Volt-Ampere characteristics and resistance result with changed temperature and wave length impact to the sample. For better accuracy measurements were done many times. All of data from measurement were cultivate by PC Easyplot programme.
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Validation of atmosphere prediction model in meso-scale
Mejstřík, Tomáš ; Žák, Michal (advisor) ; Řezáčová, Daniela (referee)
In this thesis, we are aimed at various physical principles of convective parametrization. First, traditional parametrization schemes are described. This schemes are based on hypothesis of quasi-equilibrium between large-scale processes and convective activity and suggestion about negligible mesh fraction of convection in grid-box. But, passing to resolution in the range below 10 km, the deep convection si partly subgrid, partly resolved. A new generation of prediction with high horizontal resolution is now introduced. This model has new prognostic convective scheme with new description of interaction between convective mass transport and microphysical computation of clouds and precipitation. A prognostic scheme has been used for the vertical velocity of udraft and downdraft and more elaborate microphysics, include effect of autoconversion, evaporation and sedimentation of precipitation. These models are expected to forecast many atmospheric events much more precisely (e.g. strong convection). Validation of model in experiments at different resolutions is the main task of this work. Comparsion of new developed model with model currently used, sensitivity study of new components, are presented.
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Regulace vnitřního pH kvasinek - vliv vybraných transportních proteinů
Zalom, Peter ; Chaloupka, Roman (advisor) ; Holoubek, Aleš (referee)
Intracellular pH affects nearly all biochemical processes in yeast, the processes regulating the cytosolic pH includes function of many transport proteins. In this work, the impact of selected sodium transporters on cytosolic pH has been studied in two yeast species: Saccharomyces cerevisiae and Zygosaccharomyces rouxii including wild-type and mutants with affected sodium transport. Measurements of cytosolic pH and buffering capacity have been performed using fluorescent protein probe pHluorin - a pH sensitive derivate of green fluorescence protein. Several procedures for calibration of pHluorin fluorescence response have been compared and the importance of a proper correction of the calibration curve has been demonstrated. It has been shown that cytosolic pH is influenced by the function of Nha1 transport protein in S. cerevisiae as well as in Z. rouxii but not by Sod2-22 transporter in Z. rouxii. It has been demonstrated that the buffering capacity of cytosol decrease in the presence of glucose in all strains studied.
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Sense of physical activity in patients with type 1 diabetes
Petráňová, Jitka ; Radvanský, Jiří (advisor) ; Máček, Miloš (referee)
In this thesis I'm analysing the topic physical activity with the connection to compensation of diabetes mellitus type 1. I focus on different attributes of physical activity, especially on endurance and resistance exercises. I observe the impat of physical activity on sensitivity of insulin receptors and on activation of GLUT 4 transporters in relation to the impact on compensation of diabetes. Powered by TCPDF (www.tcpdf.org)
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Study of transport proteins of the Nramp family
Surá, Lucie ; Chaloupka, Roman (advisor) ; Holoubek, Aleš (referee)
The Natural Resistance-Associated Macrophage Proteins (Nramp) form functionally conserved family of proton-dependent divalent metal ion transporters. In the present study, we investigated transport properties of a prokaryotic Nramp homolog - MntH transporter from Escherichia coli. H+ transport mediated by MntH was monitored in a bacterial model system using pH-sensitive green fluorescent protein (pHluorin). Our experimental conditions enabled us to observe an uncoupled H+ transport mediated by MntH. Uncoupled H+ flux had been previously described in eukaryotic Nramp proteins, nevertheless this is the first observation of this phenomenon in a prokaryotic homolog. We demonstrated that the uncoupled H+ transport is pH- and temperature- dependent. The uncoupled transport H+ is also affected by specific single-point mutations at functionally important residues Asp34, His211 and Asn401. The second part of the work focused on effect of different ions, which are not MntH substrates, on transport properties of MntH. It was shown that addition of excess calcium or magnesium resulted in increase of H+ transport induced by divalent metal ions, but on the other hand our data suggest that calcium inhibits uncoupled H+ transport.
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Heterologous expression of NADPH:cytochrome P450 reductase
Stráňava, Martin ; Černá, Věra (advisor) ; Martínek, Václav (referee)
NADPH:cytochrome P450 reductase (CPR) is a 78 kDa flavoprotein, which is together with cytochrome P450 component of monooxygenase system bound in the membrane of the endoplasmic reticulum. Monooxygenase system is involved in the metabolism of a wide range of organic substances, including drugs or various pollutants present in the environment (polycyclic aromatic hydrocarbons, aromatic amines, etc.). CPR works as a transporter of reducing equivalents from NADPH to the cytochromes P450. For proper interaction with cytochromes P450, intact N-terminal hydrophobic domain anchoring protein in the membrane is needed. Removing this domain, e.g. during trypsin proteolysis, gives rise a soluble CPR (72 kDa) and cause loss of catalytic activity towards cytochrome P450. During heterologous expression in E. coli proteolytically sensitive site of CPR (Lys56 - Ile57) is cleaved by intracellular trypsin-like proteases, that may negatively affect the yields of native 78 kDa protein. This thesis describes the heterologous expression, purification and characterization of two forms of rat CPR. WtCPR is a protein naturally occurring in rats (Wistar strain), while mCPR contains one amino acid substitution (K56Q) in the site of proteolytic degradation. The result of that substitution is proteolytically stable CPR,...
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Mathematical Programs for Dynamic Pricing - Demand Based Management
Hrabec, Dušan ; Karpíšek, Zdeněk (referee) ; Hoff, Arild (referee) ; Haugen, Kjetil Kare (advisor)
Tato disertační práce se zabývá vývojem, modelováním a analýzou poptávkově orientovaných úloh, které zahrnují marketingová, operační a logistická rozhodnutí. Úlohy jsou zvoleny tak, aby mohly být dále rozšířeny o koncept tzv. dynamického oceňování a jiných dynamických marketingových rozhodnutí. V práci jsou využity dvě základní poptávkově orientované úlohy: a) úloha kolportéra novin, která je zvolena pro její jednoduchou formu a která tak slouží jako nástroj pro ilustrativní ukázky rozhodovacích procesů v podobných typech úloh, a b) úloha návrhu dopravní sítě, kde jsou využity některé výsledky a znalosti získané při řešení úlohy kolportéra novin. Kolportér (či obecně maloobchodník) čelí náhodné poptávce, která může být postupně ovlivněna oceňováním, marketingovými (tj. reklamními) rozhodnutími a nakonec jejich kombinací. Poptávka obsahuje tedy náhodnou složku, která je pomocí přístupů stochastické optimalizace modelována ve specifickém tvaru (tj. aditivní či multiplikativní tvar). Závislost cena-poptávka je zachycena pomocí nelineární klesající poptávkové funkce, zatímco (vhodná) reklama vede ke zvýšení poptávky (běžně rostoucí s-křivka či konkávní funkce). Výsledky získané při řešení úlohy kolportéra novin s oceňováním jsou následně využity v úloze návrhu dopravní sítě. Tato stochastická úloha je modelována (reformulována) pomocí dvou přístupů stochastické optimalizace: wait-and-see přístup a here-and-now přístup. Jelikož tato implementace vede na lineární či nelineární celočíselnou (navíc scénářovou) úlohu, jsou v práci zmíněny taky výpočetní nástroje. Autor pro řešení používá (původní) tzv. hybridní algoritmus, což je kombinace heuristického (genetického) algoritmu a nástroje optimalizačního softwaru. Potenciální aplikace sestavených modelů, obzvláště v oblasti odpadového hospodářství, jsou diskutovány v závěrečné části disertační práce.
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