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The size of individual lipoproteins in various pathological conditions
Dušejovská, Magdaléna ; Žák, Aleš (advisor) ; Kalousová, Marta (referee) ; Kraml, Pavel (referee)
Metabolic syndrome (MS) and end-stage renal disease (ESRD) represent two clinical- pathologic states with increased risk of atherosclerotic cardiovascular complications with considerable impact on the quality of life of the patients. The knowledge about the changes in distribution of individual lipoprotein subfractions could countribute to the estimation of risk of atherosclerosis development. The studies presented in this thesis aimed at analyses of subfractions of LDL and HDL in the abovementioned pathologic states; moreover, we tried to elucidate the associations of changes in lipoprotein subfractions with clinical as well as biochemical alterations. The Study I was a placebo controlled study observing the effect of polyunsaturated fatty acids of n-3 family (PUFA n-3) administration to patients with MS who were divided to statin-treated ones (36 patients), and those without statin therapy (24 probands). The Study II comprised of 57 patients with ESRD on high volume haemodiafiltration (HV-HDF). In this Study, the parameters after 5-year follow-up were compared with baseline characteristics. Also, we included comparisons with the control group of 50 age and sex matched patients without the signs of ESRD. In Study I, we observed lowering of triacylglycerol and cholesterol content in VLDL...
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Analysis of WASH complex component FAM21
Dostál, Vojtěch ; Libusová, Lenka (advisor) ; Žárský, Viktor (referee)
The dynamics and function of the actin cytoskeleton depends on polymerization and branching of actin filaments, an event that is stimulated by Arp2/3. Arp2/3-dependent branching is closely linked to the pentameric WASH complex which consists of WASH, strumpellin, SWIP, CCDC53 and FAM21. WASH complex is associated mainly with endosomes. It was traditionally localized to retromer-coated domains of early endosomes which enable sorting and recycling of endocytosed material. However, latest scientific data extend the role of WASH complex to other endosomal or even non-endosomal sites. Of all the subunits of the WASH complex, FAM21 is the most prominent hub for protein-protein interactions, thanks to its long unstructured C-terminal domain. In my diploma thesis FAM21 was localized to early and late endosomes and lysosomes of U2OS human cell line. Dictyostelium discoideum was then used as a model organism to investigate FAM21 protein interactions as well as the proteins associated specifically with the C terminal domain of FAM21. Results of the study shed new light on the complex network of FAM21 interactions and question the long-standing theories on the function of WASH complex in cells. Powered by TCPDF (www.tcpdf.org)
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Large laser sparks for laboratory simulation of high-energy-density events in planetary atmospheres
Babánková, Dagmar ; Civiš, Svatopluk (advisor) ; Strauch, Bohuslav (referee) ; Špirko, Vladimír (referee) ; Rohlena, Karel (referee)
3 Conclusion The main goal of this doctoral thesis is to investigate the possibility of synthesis of simply organic molecules from inorganic gases such CO,CO2N2,NH3,CHa,H2O,H2, using the large laser sparks. The laser sparks provide a unique way to mimic the chemical effects of high-energy-density events in planetary atmospheres (cometary impact, lightning) matching the natural energy-density, its spatio-temporal evolution and plasma-volume scaling ofsuch events in a fully-controlled laboratory environment. Laser induced dielectric breakdown was induced by Š1kJ laser pulses in molecular gases and mixtures related to various planetary atmospheres. We believe that the single-shot experiments realized at the high-power laser facility simulate more realistically high-energy-density atmospheric events than similar experiments conducted with electrical discharges or experiments using low- energy pulses from high-repetition-rate lasers. In our case three different gaseous mixtures were used for these purposes; the strongly.reduced mixture NH3-Cř{4.H2-H2o, medium-reduced mixture CO-N2-H2O and mildly-reduced mixture CO2-N2-H2O. For all kind of my experiments the emission spectra w€re measured in different spectral regions. The motecular bands of CN and C2 radicals dominates in the visible spectra taken from the...
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Modified Substrates in β-N-Acetylhexosaminidase-Catalyzed Synthesis
Bojarová, Pavla ; Křen, Vladimír (advisor) ; Moravcová, Jitka (referee) ; Walterová, Daniela (referee)
4 Conclrrsion 4 CoNcrusroN This Ph. D. thesis is a systematic study of the substrate specificity and the synthetic potential of B-N-acetylhexosaminidases (EC 3.2.1.52) with structurally modified substrates. It comprises four publications in intemational journals, one review and 17 oral and poster contributions. The following parts of the substrate molecule were modified: 2-acetamido moiety, the C-6 hydroxyl (oxidations, introduction of a cyano group) and the aglycon part (glycosyl azides - C-N bond hydrolysis). Thirteen modified substrates were synthesized, seven of them were described for the first time. They were tested for hydrolysis and transglycosylation by over thirty fungal $-N-acetylhexosaminidases (culture collections at Charles University and at the Institute of Microbiology, Academy of Sciences of the Czech Republic) and the results were discussed in relation to the conclusions of molecular modeling (B-N-acetylhexosaminidase from Aspergillus oryzae CCF 1066). Eight oligosaccharidic structures (six of them novel) were prepared by semi- preparative transglycosylation reactions (tens of miligrams), isolated (mostly 16_377a yields, even 787o yie\ď) and fully characterized. Noteworthy properties like immunoactivity (binding to natural killer cell activation receptors) and inhibitory potential were...
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Role of membrane phospholipid remodeling, oxidative stress and PKC-δ upregulation in cardioprotection induced by chronic hypoxia
Ježková, Jana ; Kalous, Martin (referee)
Role of membrane phospholipid remodeling, oxidative stress and PKC- δ upregulation in cardioprotection induced by chronic hypoxia Introduction: Cardiovascular disease is the main cause of morbidity and mortality in industrially developed countries. Therefore, a number of experimental and clinical studies deal with the issue of how to increase tolerance to myocardial ischemia/reperfusion (I/R) injury. The present studies are concerned with mechanisms involved in the induction of endogenous cardioprotection after adaptation to intermittent altitude (IHA) hypoxia and influence of fat diets enriched with different fatty acids (FA) composition. We focused on changes in the composition of serum and heart lipids, the expression of PKC and on markers of oxidative stress and antioxidant enzyme activity in the myocardium. We also studied the role of reactive oxygen species (ROS) in the cardioprotective effect of IHA hypoxia using the antioxidant N-acetylcysteine (NAC). Methods: Adult male Wistar rats were exposed to IHA hypoxia in hyperbaric chamber, altitude 7000 m, 8 hours per day, 5 days a week, 25 exposures. Animals were fed by standard diet and diet enriched with 10% fat (saturated - SFA and MUFA mononemnasycené), 10% corn oil (n-6 polyunsaturated MK PUFA) and 10% fish oil (n-3 PUFA) in the diet experiment....
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Efficiency of chemical stabilization of a contaminated soil using amorphous manganese oxide
Tomášová, Zdeňka ; Ettler, Vojtěch (advisor) ; Trakal, Lukáš (referee)
Soil contamination by toxic metals and metalloids is currently one of the most discussed topics in the environmental sciences. Chemical stabilization of contaminated soils using amendments such as Fe, Mn and Al oxides belongs to one of the in situ remediation methods. Manganese oxides are naturally present in soils and they exhibit a number of properties for being efficient sorbents of toxic metal(loid)s, such as a low point of zero charge, a large specific surface and a structure that allows ion acceptance. The focus of this thesis is to determine efficiency of chemical stabilization of a smelter-contaminated soil using the amorphous manganese oxide (AMO) under various pH conditions. The agricultural soil polluted by emission from a Pb smelter (located in Příbram, Czech Republic; Pb 1 100 mg/kg, Zn 294 mg/kg, Cd 4.98 mg/kg, As 118 mg/kg and Sb 48.9 mg/kg) was amended with the AMO and incubated for 2 and 6 months. The soil was subsequently subjected to a pH-static leaching procedure in the pH range of 3-8. The presence of AMO in soil increased the soil natural pH from 5.77 to 6.59 and 6.23 after 2 and 6 months, respectively. The pH-static experiments indicated that no effect of the AMO treatment was observed for Cd and Zn, whereas the leaching of other contaminants (As, Cu, Pb, Sb) decreased...
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Časoprostorové vzory zmlazení v přírodních temperátních lesích
Holík, Jan
Interactions among woody individuals can be mediated by natural enemies depending on the local community composition, thus contributing fundamentally to the formation of the spatio-temporal patterns of tree species in temperate forests. The effects of density and distance to neighbouring individuals are thought to influence the susceptibility of woody individuals to predation and their survival. However, how the effects of neighbouring individuals depend on, for example, the species and life stage of woody individuals, type of natural enemies and environmental conditions is poorly understood. The spatial extent of the effects of neighbouring individuals that influence the susceptibility of woody individuals to predation and their survival also remains an important question. This dissertation addresses the role of neighbourhood effects on the predation (herbivory) and survival of woody individuals at early life stages at several temperate forest sites in the Czech Republic. It compiles the findings from one experimental and three observational studies representing its main research areas. In terms of ecological theory, the area of research on woody seedling survival investigates neighbourhood effects associated with the Janzen-Connell effect (Papers 1, 2). In terms of foraging theory, the area of research on the susceptibility of woody individual to herbivory then investigates associational effects (Papers 3, 4). The dissertation evaluated variation in spatial scale and direction of neighbourhood effects on seedling survival in a lowland alluvial forest depending on seedling size, growth form and species (Paper 1). We found a negative relationship between the density of conspecific neighbours and seedling survival only at low light availability. This relationship was observed for the group of all seedling species except the most abundant Acer campestre. The next part of the dissertation examined the importance of natural enemies for the neighbourhood effects of adult trees (Paper 2). Due to vertebrates and partially insects, the survival of first-year Fagus sylvatica seedlings was lower beneath conspecific than heterospecific adults and it was also lower beneath conspecific adults than the survival of Picea abies. Conversely, the establishment of Picea abies was lower beneath heterospecific than conspecific adults, although it was eventually lower beneath conspecific adults than the establishment of Fagus sylvatica. The last part of the dissertation evaluated spatial patterns in neighbourhood effects on deer herbivory (Papers 3, 4). A positive relationship between conspecific neighbours and browsing of Fagus sylvatica seedlings was significant only at short distances up to 1.5 m. Conversely, we found that Abies alba neighbours increased browsing of Abies alba and Fagus sylvatica seedlings along the whole distance interval up to 4 m. In a species-rich lowland alluvial forest, neigbourhood effects of trees and shrubs either increased or decreased the susceptibility of individuals to bark stripping and its intensity at distances of 2–30 from neighbouring individuals depending on the species of focal and neighbouring individuals. Finally, I illustrate that the local composition of tree communities can modify the susceptibility of woody individuals to herbivory and their survival at early life stages. I identify the species, life stage of focal and neighbouring individuals, type of natural enemies and light availability in the understorey as key factors determining the importance of neighbourhood effects. This dissertation also shows how and at what spatial scales enemy-mediated neighbourhood effects can contribute to community structure and composition.
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Analýza imunitní odpovědi na infekci Actinobacillus pleuropneumoniae
Jarošová, Rea
Actinobacillus pleuropneumoniae (APP) is a specific respiratory pathogen and etiological agent of swine pleuropneumonia, a highly contagious disease that causes significant global economic losses, despite the availability of commercial vaccines against the disease. The experimental part of this work is conceived from two partial points of view: The first, dealing with the investigation of nonspecific immune mechanisms, and the second, dedicated to the immunization of pigs against APP. The primary goals were to optimize the method of cryopreservation of lung tissue and subsequently, using immunohistochemical analysis together with the innovative MALDI MSI method, to detect inflammatory mediators in the tissues of infected pigs. The main objective was to determine, whether specific populations of CD163 positive inflammatory monocytes and – or macrophages are responsible for the production of these inflammatory mediators. The second main objective was to compare the ability of different adjuvants to modulate the immune response in relation to the route of vaccine administration and to evaluate these findings in relation to adverse local reactions and the protection of pigs in the challenge test. The results showed that: 1) the most suitable method of cryopreservation of lung tissue is the instillation of a mixture of cryoprotectant O.C.T. with phosphate buffered saline 1: 1 into the lung parenchyma and freezing in supercooled nheptane placed on dry ice; 2) spatial distribution of inflammatory mediators in the tissues of infected pigs can be detected by MALDI MSI and this method can be used to supplement the current methods; 3) inflammatory CD163 positive monocytes are unlikely to contribute to elevated plasma cytokine levels. However, they are source of inflammatory mediators, especially interleukin 1beta at the site of inflammation; 4) intradermal administration of vaccine with the mineral oil adjuvant Montanide ISA 201 VG (Seppic, France) is more reactogenic and slightly less immunogenic; however, it provides more effective protection in the challenge test than intramuscular administration of the vaccine.
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Porovnání různých systémů hnojení v podmínkách ekologického zemědělství
Antošovský, Jiří
The main treatment to ensure a sufficient nutrient in organic farming is realized by green manure crop cultivation, post-harvest residues incorporation to the soil and application of organic fertilizers. The aim of this long-term experiment was to compare two different production systems with and without livestock in terms of organic farming. The idea of this long-term experiment is to determine whether an organic farming can be sustainable without livestock production or not. Control variant with no fertilization (treatment 1) and incorporation of sole green manure crops (treatment 2) were also included. The production system without animal husbandry was based solely on the application of renewable external resources (compost or digestate) (treatment 3) and the same fertilization with the addition of auxiliary substances (AS) (treatment 4). The production system with animal husbandry included utilization of fertilizers produced on the farm (fermented urine or manure) using solely farm fertilizers (treatment 5) and in addition with AS (treatment 6). Each treatment had three replications. This work describes the average yields from four experimental years and five localities. Winter wheat, potatoes, winter wheat spelt and legume-cereal mix with corn were used and examined as model crops during the first four years of this long-term research. The highest average yield of winter wheat grain and potato tubers during the first two years of the experiment were obtained after the treatments 3 (7.1 t/ha grain, 33.9 t/ha tubers) and 4 (7.0 t/ha grain, 34.1 t/ha tubers). The several times higher nitrogen content in applied digestate and compost in addition with narrow C:N ratio of applied compost in comparison with fermented urine and manure were probably the reasons for such results. On the contrary, the results obtained from the third (spelt) and fourth (LCM and corn) experimental years was higher after treatment 5 (5.5 t/ha grain, 4.6 cereal unit/ha) and 6 (5.4 t/ha grain, 4.7 cereal unit/ha). After four experimental years, the presented results supported the application of farm fertilizers as a preferable option from the long-term point of view. The treatments with additional application of AS did not provide a higher yield, therefore, such an application seems unnecessary. The average yield after the fertilization with solely green manure crops was higher in comparison with control variant, but it was statistically insignificant.
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Effect of selected abiotic stress factors on yield formation parameters of winter wheat genotypes
Hlaváčová, Marcela
This dissertation thesis is a compilation of four scientific papers based on experiments with pot-grown winter wheat plants. The responses of winter wheat to different lengths of drought and heat stress treatments, applied at growth phases crucial for grain formation, both from the perspective of final yield formation and photosynthetic parameters, were analyzed. Growth chambers (heat and drought stress) and phenotyping platform (drought stress; phenotyping experiment) were used to set up these stress conditions. The first aim was to assess the sensitivity of yield formation parameters to short-term heat stress at anthesis. The second aim was to assess the sensitivity of yield formation parameters to drought stress during the transition from the vegetative stage to the generative stage. The third aim was to assess the sensitivity of yield formation parameters and photosynthetic characteristics to combined short-term heat and drought stress at stem elongation, anthesis, and grain filling stages. The final aim was to assess the sensitivity of photosynthetic characteristics to combined medium-term heat and drought stress at the anthesis stage. The results showed that short-term (3 and 7 days) heat (32, 35, 38 °C, and compared to a control temperature of 26 °C) or interactive effects of heat and drought stress at stem elongation, anthesis, and grain filling stages generally led to minor reductions in yield formation parameters, whereas significant affects on photosynthetic parameters were observed (with the lowest impacts on photosynthetic parameters at anthesis). It indicated that plants were capable of quickly recovering their photosynthetic processes; thus, yield formation parameters could be compensated. Drought stress during the transition from the vegetative stage to the generative stage of development resulted in significant declines in yield formation parameters. The photosynthetic responses to medium-term (7 and 14 days) combined heat and drought stress were genotype-specific in their magnitudes and closely associated with changes in stomatal conductance.
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