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Application Database Layer Generator
Kuboš, Jaroslav ; Rychlý, Marek (referee) ; Burget, Radek (advisor)
This diploma thesis deals with the design and implementation of a framework for the database persistence layer development. This framework is easy to use while keeping the code elegance. It supports object oriented programming features such as inheritance and collections. Other features include versioning of objects and lazy loading. The object metadata are obtained through reflection provided by the .NET framework. The framework is not using any literal for identification (classes, attributes) even in object queries. Most of checks are done by compiler.
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The Study of Distribution of Halogenated Diphenylethers to Parts of Environment
Hroch, Martin ; Hajšlová, Jana (referee) ; Kráčmar, Stanislav (referee) ; Šucman, Emanuel (referee) ; Vávrová, Milada (advisor)
In recent years considerable attention is paid to problems of brominated flame retardants (BFR's – Brominated Flame retardants), which are a diverse group of organic compounds. Even in the recent past have been the most often represented group particularly polybrominated diphenyl ethers (PBDEs), which have found wide use in many industrial sectors. The reason of aplication of these substances is the ability to slow down the combustion process and reduce the risk of ignition by the excessive heating of material. On the other hand, serious concerns about the use of PBDEs was added. The most serious trouble is particular their possible to release from consumer products during their normal use, toxicity and high persistence in parts of environment. Some of these contaminants are more characterized by a high degree of accumulation in biological systems. In this work by several sub-studies the issue of assessment the level of pollution of the aquatic ecosystem of selected localities of the Czech Republic just PBDEs was addressed. One of objectives was monitored the situation of breeding ponds near the village Záhlinice in Central Moravia. Here was assessed level of contamination in fish and birds as two consecutive segments of food chain. The obtained results largely confirmed the assumption that with increasing trophic level of organisms leads to bioaccumulation and thus also higher findings. For comparison, the capture of fish and birds in other localities of the CR was also implemented. In Bartošovice and Hustopeče nad Becvou levels at the buzzard and heron were detected as comparable. The cormorants of the order higher levels of PBDEs were found. Further were also evaluated levels of PBDEs detected in individuals of bioindicator kind of chub caught in two locations on the river Svratka in spring and autumn. Findings of PBDE in muscle, skin and intestines were compared. The obtained results show that between tissues and sites wasn’t significant variability respectively and the findings of controlled substances are practically comparable. With aim to determine the dependence of the concentration of PBDEs in organisms on the length of the river was conducted monitoring of Vír and Brno water reservoirs. In both locations was main monitored kind of bream, which was completed by the other species. The results of the analysis indicate a higher total concentration at the lower part of river Svratka. Differences were also the distribution of individual congeners in both sampling locations. For monitoring of terrestrial system contamination levels of PBDEs has been selected several types of plant bioindicators. These are mainly pine needles (Scots pine, Eastern white pine, Silver fir, Blue spruce) collected from different localities of the Czech Republic. Other matrices were chosen roughages (Common wheat, Naked barley, Alfalfa wheat, Red clover) and oilseeds (Rapeseed) from the Novy Jicin, where Agriculture plant school of the University of Veterinary and Pharmaceutical Sciences Brno. Detected values of PBDE concentrations were very low, generally ranged at the detection limits of the analytical method.
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Influence of Staphylococcus aureus Quorum sensing system to antibiotic tolerance
Šaňková, Michaela ; Lichá, Irena (advisor) ; Buriánková, Karolina (referee)
Staphylococcus aureus is a gram-positive opportunistic pathogenic bacterium that causes a range of chronic infections especially to weakened individuals. Recently, the most serious problem in clinical medicine has been infections caused by so-called persisters. They form a subpopulation of cells that, thanks to adaptations to stressful conditions, are able to withstand lethal doses of antibiotics and then cause an infection again. Agr system, which is responsible for the mechanism of quorum-sensing, plays the main role in switching between the active virulent form and the persistent form associated with biofilm formation. During chronic infections, adaptive mutations accumulate in this Agr system and affect the ability to persist. For the purpose of this thesis, clinical isolates were obtained from patients diagnosed with cystic fibrosis in chronological order within the first three years of their lives. The aim of this thesis was to confirm the influence of adaptive mutations on the persistence ability of clinical isolates and also to observe how the membrane potential and metabolic activity change depending on the on the action of selected antibiotics. Additional aim was to create a deletion of the response regulator AgrA gene using the CRISPR/Cas9 system. In order to characterize the persistence...
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Effect of adaptive mutations on persister formation in Staphylococcus aureus
Hloucalová, Nikola ; Lichá, Irena (advisor) ; Buriánková, Karolina (referee)
Persistence represents a transient state during which are bacterial cells able to survive antibiotic treatment. Only a small subpopulation of cells enter this state, these cells are then capable of causing disease recurrence. The size of a persister subpopulation is influenced by adaptive mutations formed during a chronic disease in genes related to transition of cells between a virulent form and biofilm formation. Regulation of entering and exiting persistence is also influenced by intercellular communication and by molecules produced into the cell surroundings. It is assumed that this effect is caused by PSM modulines and AIP molecules which take part in Quorum sensing mechanism. The main aim of this thesis was to clarify how adaptive mutations of clinical isolates obtained from two patients diagnosed with cystic fibrosis influence the ability to persist. Another aim was to determine changes in membrane potential and metabolic aktivity while entering persistence and to establish the effect of extracellular molecules produced into the culture medium on growth parameters of the studied isolates. By using techniques of TD test and establishment of persistence rate by CFU determination, it was found that the isolates obtained from patient 2 did not acquire adaptive mutations affecting persistence...
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Methods for characterization of persistent state after exposure to selected antibiotics in Staphylococcus aureus
Valtová, Aneta ; Lichá, Irena (advisor) ; Tkadlec, Jan (referee)
Staphylococcus aureus is a opportunistic pathogen that can cause severe and chronic infections. The reason of the infections relapse is often the persistence. It is about adapting to stressful conditions by inducing a dormant state, which would allow bacteria to survive exposure to antibiotics and grow again after their elimination. Bacteria that persist in the patient acquire various adaptive mutations, which are transmited creating subpopulations that have a better ability to persist. The aim of this diploma thesis was to compare individual methods of persistent study that could be used in clinical practice in the future, and at the same time to try a closer molecular characterization of the persistent state with using methods for calculating gene expression. I had chronological isolates of Staphylococcus aureus at my disposal, the initial one being the primoisolate, an isolate taken at the diagnostics of cystic fibrosis before the start of antibiotic treatment. Another was taken at a distance of three-quarters of a year and the last with a half-year interval from the previous one. Following whole genome sequencing, genes in which adaptive mutations occurred were identified. The first method determines the degree of persistence by calculating CFU (Colony Forming Units) after antibiotic treatment....
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The Persistence of Human Polyomaviruses
Blažková, Kristýna ; Drda Morávková, Alena (advisor) ; Kuthan, Martin (referee)
Despite years of research, even the most scrutinized Polyomaviruses - BK and JC - have not yet been thoroughly understood. With a number of new Polyomaviruses - KIV, WUV, MCV, HPyV6, HPyV7, TSV and HPyV9 described in the past few years, the need to understand how Polyomaviruses operate in their hosts has become even more urgent. The probable route of transmission appears to be either respiratory or faecal-oral. The initial infection occurs most likely in the early childhood or early-adolescence and is followed by a life-long persistence. The seroprevalence of Human Polyomaviruses among healthy adult population is high: BKV (81-97 %), JCV (35-69 %), KIV (55 %), WUV (69 %), MCV (25-46 %) and TSV (70-80 %). Human Polyomaviruses can cause fatal diseases in immunocompromised patients. The site of persistence in humans probably varies depending on the specific Polyomavirus. BK and JC are known to persist in kidneys and the urinary tract. Human Polyomaviruses have been detected in the lymphatic tissues, blood, respiratory, urinary, and gastrointestinal systems. It is not clear, however, if they persist in all of these sites. Mechanisms which Polyomaviruses use to establish and maintain persistent infection could include the viral miRNA and viral agnoprotein, which would result in a modulation of viral...
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Adherence to pharmacotherapy of osteoporosis
Santariová, Martina ; Vytřísalová, Magda (advisor) ; Doseděl, Martin (referee)
ADHERENCE TO PHARMACOTHERAPY OF OSTEOPOROSIS Author: Martina Santariová Supervisor: PharmDr. Magda Vytřísalová, Ph.D. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Social and Clinical Pharmacy INTRODUCTION: Adherence is the extent to which the patient's behavior (using of medication, following recommended regimen) is in accordance with the advice of the health care provider. Adherence to pharmacotherapy of osteoporosis, including compliance and persistence, is still suboptimal. OBJECTIVE: The objective of the study was assessing of subjectively evaluated adherence with peroral bisphosphonates used to treat osteoporosis within Czech women. METHODS: A cross-sectional multicentre survey. Data collecting was carried out using anonymous questionnaire in five osteocenters in the Czech Republic. The research was performed between November 2012 and March 2013. Adherence was evaluated using a standardized questionnaire OS- MMAS (Osteoporosis-specific Morisky Medication Adherence Scale). RESULTS: A total of 363 postmenopausal women aged 55 years and older with osteoporosis or osteopenia were participated in the study. 133 (36,6 %) women used weekly dosage forms of BIS (alendronate, alendronate and vitamine D in a fixed combination, risedronate) and 230 (63,4 %) women used...
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Interaction of Borrelia sp. with HL-60 cells and monocytes and cultivation of Anaplasma phagocytophilum in HL-60 cell culture
Marková, Lucie ; Hulínská, Dagmar (advisor) ; Beranová, Jana (referee)
Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...
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Mechanisms of bacterial persistence and clinical significance
Smělíková, Eva ; Melter, Oto (advisor) ; Pinkas, Dominik (referee)
Persistent bacteria, shortly persisters, are cells that are characterized by their tolerance to antibiotics without containing resistance genes. These are not resistant bacteria, because resistant bacteria are determined by genetic code. Persisters are indistinguishable from other antibiotic sensitive bacteria, and they are rather transient phenotypic subpopulations. Probably all types of bacteria can create a persistent stage, the ability is not species-bound and persistence has been described in a number of bacterial species. The reason for the formation of persistence may be sudden stress, then it is induced formation, or the reason may be an insurance for the future, then we call it stochastic phenotypic variability. Then a variety of phenotypes of different subpopulations within a genetically uniform population can be distinguished. They differ in growth properties and tolerance to antibiotics. Bet-hedging strategy is a hypothesis that describes persistence as insurance against the risk of external fluctuations. During stress an alarmon guanosine tetraphosphate or guanosine pentaphosphate is accumulated to alert the cell to unfavorable conditions and it initiates transition to a more tolerant state. The existence of these tolerant subpopulations is important in recurrent diseases because small...
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