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Laboratory diagnosis of mycobacteria, with a focus on the bacteriological proof
VELKOVÁ, Martina
Laboratory methods play an important role in disease diagnostic. Although there are currently available fast molecular genetic methods and methods based on cell response, basic diagnostic method of direct evidence is still microscopy and cultivation. Microscopy (staining by Ziehl-Neelsen and fluorescence microscopy) is the primary method for most of the specimens, especially for sputum. The sensitivity of this method is not very high because for the detection of a positive finding in 1 mm3 is needed at least 105 microbes. However, the method is important because it can fastly proof extensive clinical and epidemiological serious diseases. The ?Golden standard? is today the cultivation on solid egg media. Nowadays it is filled in automatic detection system, which accelerates the time of detection. The thesis was carried out in the Laboratory of Medical Microbiology, Bacteriology department, Hospital Ceske Budejovice in the period from 1.1.2013 till 6.6.2013, and included 400 sputa, which were processed and examined. The purpose was to compare the effectiveness of decontamination methods with HCl and with NALC, and to monitor the recovery and detection for the automatic detection system and conventional cultivation. In the automatic system using the BACTECTM MGITTM 960 Non-Radiometric fluorescent technology is cultivation in liquid Middlebrook 7H9 medium supplemented with antimicrobials (polymyxin B, amphotericin B, nalidixic acid, trimethoprim, azlocilin). The classical culture used solid egg culture medium: Lowenstein-Jensen and culture medium by Ogawa. The individual results then show that the method of HCl had the overall contamination of 1 % of the samples. For each methods in 5 % BACTECTM MGITTM 960 and 4 % in the conventional cultivation. In comparison the method NALC, had the overall contamination of 21 % of the samples. For the individual methods in 26 % of BACTECTM MGITTM 960 and 50 % in the conventional cultivation. From these results it is obvious that the method with the NALC has lower efficiency, the proportion of contamination compared to the method with HCL increased by 20 %. Decontamination with the NALC method recommended by the manufacturer for BACTECTM MGITTM 960 was found to be unsatisfactory and was canceled. Comparing the recovery and the detection of the strains isolated in individual methods showed that the BACTECTM MGITTM 960 exhibits greater sensitivity than conventional cultivation, since the total of 17 strains isolated BACTECTM MGITTM 960 captured 15 against 10 strains isolated in conventional cultivation. There is also a significantly shorter time to detect positive samples. The average detection time for BACTECTM MGITTM 960 was 16.2 days, while a conventional culturing was 31 days. It was confirmed that the BACTECTM MGITTM 960 system achieves better results, but optimization is achieved by combining the two methods used. There is an apparent decrease in detection of mycobacteria from the processed statistical data of the strains isolated during the period 2010 - 2012, but a substantial reduction does not occur. Noticeable is only the decline of the isolated stains of Mycobacterium bovis BCG, which can be explained by the fact that the vaccination against tuberculosis is since 2011 no longer carried out across the board, but according to the new legislation in force only in high-risk groups. An interesting fact is that every year the highest laboratory detection is found in the age group of over 60 years.
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Analysis of metabolome effects of hop (H. lupulus) transcription factors in heterologous of Petunia hybrida
MORAVCOVÁ, Vendula
Plant transformation is now a key research in plant biology, and also is a unique practical tool in the improvement of varieties of cultivated plants. The aim of this work was to obtain transgenic plants Petunia x hybrida containing transgene nptII (kanamycin resistance gene) using the indirect transformation by Agrobacterium tumefaciens. In terms of in-vivo was grown from seeds experimental material in the form of plant Petunia x hybrida cv. Andrea. For the experiments three different constructs HLWD40 3278-80 and HLbHLH 3577 and HLbHLH 3677 GFP were used. The leaf explants from Petunia plants were prepared. Explants were transferred using tweezers into the crucible containing liquid ? MS and prepared bacteria were added. Thus prepared explants remained in bacteria the next day, when they were transferred to the regenerating solid ground. The explants formed callus gradually (clusters of cells) which regenerate a new plant. Transformation should satisfy the condition that the transformed plants should be fertile. For this reason, an attempt was made crossing the transformed plants were crossed Pap1 1527/2 and Pap1 1572/4 with plants crossed HLWD 40 3278-80. The grown plants were analysed by a PCR reaction. The explants began during the first 2-4 weeks after transfromation to form callus and during the next week to remove the first regenerants. At one exulant fell averaged around 7 regenerants and the explants regenerated by construct HLbHLH 3577 regenerate better than regenerants transformed with construct HLWD 40 3278-80. During the transformation we obtained regenerants. With "tissue" PCR, it was found that 15 plants carried the gene for kanamycin resistance. Transformation efficiency was 3.75%.
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Company Investment Strategy
Svoboda, Filip ; Krátký, Petr (referee) ; Čižinská, Romana (advisor)
The diploma thesis deals with the issues of a company´s investment strategy. The aim is to propose an investment portfolio that will meet the company requirements regardless of its field of business, efficiently utilize various financial instruments and, at the same time, meet the requirement of liquid money.
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Localisation of Fluorescent Probes and the Estimation of Lipid Nanodomain Sizes by Modern Fluorescence Techniques
Šachl, Radek ; Hof, Martin (advisor) ; Obšil, Tomáš (referee) ; Almgren, Mats (referee)
iii Organizations Umeå University Department of Chemistry, Umeå, SE & Charles University in Prague, Faculty of Science, Prague, CZ Document name Doctoral thesis Date of issue February 2012 Author Radek Šachl Title Localisation of Fluorescent Probes and the Estimation of Lipid Nanodomain Sizes by Modern Fluorescence Techniques Abstract The thesis is divided into two major parts. The first part focuses on the localisation of probes in lipid/polymeric bilayers and in GM1 micelles. Included in this thesis is a new approach based on electronic energy transfer/migration (FRET/DDEM), which efficiently determines transversal positions of fluorescent molecules in lipid bilayers. This approach has been used to locate newly synthesized lipid probes in DOPC bilayers. The label was introduced at the end of sn-2 acyl chains of variable length. Analytical models accounting for FRET exist for a limited number of basic geometries. Here, a combination of FRET and Monte Carlo simulations enables the localisation of probes in bicelles and in bilayers containing pores, i.e. in lipid systems with variable curvature, or in non-homogenous lipid systems. This approach has been used to test whether conical-like fluorescence probes have an increased affinity to highly curved regions, which would enable preferential labelling of...
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Liquid phase epitaxy
Nohavica, Dušan
Extended lecture collects mojority aspect of the "Liquid Phase Epitaxy", LPE, with special attention to the semiconductors growth. Methods of growth modifications enabling to increase growth reproduction and deposition efficiency are included. Also dislocations generation by different mechanisms is discussed in the article.
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The effectiveness of welding on furnace structures.
Rousová, Michaela ; Mrňa, Libor (referee) ; Kubíček, Jaroslav (advisor)
This diploma thesis is resolving all the possibilities of enhancing the efficiency of furnace structures welding. Small batch production does not offer many possibilities for implementing mechanization or automation. On the other hand, when using a big batch production, efficiency can be enhanced by means of a robotic workstation. When the production batch is big enough, we will see a costs save after a short time, mainly in labor costs. This means the return of investments will be in short time period. In the LAC company there are three types of products made. At laboratory furnaces the efficiency can be enhance by using fixtures. At other standard and atypical furnaces is very difficult to design fixtures or positioners because of their different sizes. Big complication can be also a company location on second floor. The most suitable product regarding to welding efficiency enhancement is big batch production of heaters. For this type the welding time can be shortened by means of a robotic workstation.
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Modern reverse stationary phases based on silicagel, zirconium dioxide and organic monoliths: application to analysis of biologically active compounds
Soukupová, Klára
Charles University in Prague Faculty of Science Department of Analytical Chemistry Modern Reversed Stationary Phases Based on Silicagel, Zirconium Dioxide and Organic Monoliths; Their Used in Separation of Biologically Active Compounds Synopsis of PhD. Thesis Prague 2008 Klára Soukupová The PhD. Thesis was carried out at the Department of Analytical Chemistry, Faculty of Science, Charles University in Prague, in the period of 2003-2008. Author: Mgr. Klára Soukupová Supervisors: RNDr. Jana Suchánková-Sobotníková, Ph.D Doc. RNDr. Eva Tesařová, CSc. Prague 2008 1 Table of contents 1. Introduction 2 2. Objectives of the Thesis 3 3. Results and Discussion 4 4. Conclusions 12 5. References 13 Appendix 1: List of Publications, Lectures and Poster presentations 14 Appendix 2: Curriculum Vitae 16 2 1. Introduction High performance liquid chromatography (HPLC) is modern instrumental method used mainly in analytical chemistry. Most used separation mode is reversed phase liquid chromatography (RP HPLC). This method is very useful because of high separation efficiency, reliability and wide choice of stationary phases with different selectivity. Most used stationary phases in RP HPLC are bonded phases, which have many advantages: good availability, wide application range, fast establishment of equilibrium and ability to...
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Modern reverse stationary phases based on silicagel, zirconium dioxide and organic monoliths: application to analysis of biologically active compounds
Soukupová, Klára ; Sobotníková, Jana (advisor) ; Pacáková, Věra (referee) ; Cvačka, Josef (referee)
Charles University in Prague Faculty of Science Department of Analytical Chemistry Modern Reversed Stationary Phases Based on Silicagel, Zirconium Dioxide and Organic Monoliths; Their Used in Separation of Biologically Active Compounds Synopsis of PhD. Thesis Prague 2008 Klára Soukupová The PhD. Thesis was carried out at the Department of Analytical Chemistry, Faculty of Science, Charles University in Prague, in the period of 2003-2008. Author: Mgr. Klára Soukupová Supervisors: RNDr. Jana Suchánková-Sobotníková, Ph.D Doc. RNDr. Eva Tesařová, CSc. Prague 2008 1 Table of contents 1. Introduction 2 2. Objectives of the Thesis 3 3. Results and Discussion 4 4. Conclusions 12 5. References 13 Appendix 1: List of Publications, Lectures and Poster presentations 14 Appendix 2: Curriculum Vitae 16 2 1. Introduction High performance liquid chromatography (HPLC) is modern instrumental method used mainly in analytical chemistry. Most used separation mode is reversed phase liquid chromatography (RP HPLC). This method is very useful because of high separation efficiency, reliability and wide choice of stationary phases with different selectivity. Most used stationary phases in RP HPLC are bonded phases, which have many advantages: good availability, wide application range, fast establishment of equilibrium and ability to...
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IEF and HPLC-based methods for efficient bacterial characterization
Kubesová, Anna ; Šalplachta, Jiří ; Moravcová, Dana ; Horká, Marie
Pectobacterium and Dickeya species are responsible for blackleg and tuber soft rot diseases on crop and ornamental plants. Recently, a more virulent and aggressive Dickeya type of E. chrysanthemi was described and it becomes serious problem for potato production in Europe. In this study, we suggest procedures using two commonly available laboratory techniques, gel isoelectric focusing and liquid chromatography, for differentiation and characterization of Pectobacterium and Dickeya species. We have confirmed that fingerprinting approach can be used for bacterial differentiation, which can be further used in rapid diagnosis of plant disease.
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Silica-based monolithic capillary columns modified to zwitterionic stationary phase for hydrophilic interaction liquid chromatography
Moravcová, Dana ; Planeta, Josef ; Kahle, Vladislav ; Horká, Marie ; Roth, Michal
Zwitterionic monolithic capillary columns intended for isocratic gradient hydrophilic interaction chromatography (HILIC) separations are introduced. Silica-based capillary columns (150 mm x 0.1 mm) were prepared by acidic hydrolysis of tetramethoxysilane in the presence of polyethylene glycol and urea. The modification by a 3-(trimethoxysilyl)propyl methacrylate and then by a zwitterionic [2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide to HILIC stationary phase bearing sulfoalkylbetaine groups on its surface followed. Prepared columns were characterized in HILIC separation mode employing mobile phase containing 10% (v/v) of 5 mM ammonium acetate pH = 4.5 in acetonitrile. Comparison with the commercially available ZIC-HILIC® column (Merck SeQuant®) under the same separation conditions using a mixture of aromatic carboxylic acids as a sample was done on the basis of separation efficiency of tested columns as well as retention factors and peak asymmetry of individual solutes.
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