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Plasmid DNA vaccines
Machan, Radoslav ; Chroboková, Maria (referee) ; Rittich, Bohuslav (advisor)
Plasmid DNA vaccines are the new generation of vaccines with a great potential in prevention of many diseases. Recent studies and clinical test are aimed at prevention against cancer, hepatitis, malaria, HIV, influenza and other diseases. Recent main challenges covering plasmid DNA vaccines are associated with optimalization of each step of production and mainly purification steps allowing production of pDNA at kilogram levels. Main purification techniques used are based upon chromatographic methods, but research and development also shows other potential methods, like two-phase aqueous systems or magnetic microparticles as carriers. In experimental part of this thesis isolation of pUC 19 plasmid from Escherichia coli JM 109 (pUC 19) cell culture was performed via method of alkaline lysis. Isolation was verified by agarose gel electrophoresis. Isolated samples were purified in four repetitions with lithium chloride and magnetic microparticle carriers and the extent of purification was verified spectrophotometrically. Purified samples were visualised via agarose gel electrophoresis and results were compared.
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Nucleic acids as terapeutic agent
Ráčková, Lucie ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
With the development of molecular biology development of oncotherapy proceeds. The major progress of modern medicine is gene therapy. In the gene therapy are two categeories, namely, viral vectors and nonviral vectors which are used mainly. Nonviral vectors include plasmids. Plasmid DNA used in medicine must be perfectly purified. Chromatographic methods are mainly used at present. Research and development deals with other methods for example two-phase aqueous systems and magnetic carriers. In experimental part of this thesis, isolation of pUC 19 plasmid DNA from Escherichia coli JM 109 (pUC 19) cell culture was performed via method of alkaline lysis. Quality of isolated plasmid DNA was verified spectrophotometrically and by agarose gel electrophoresis. Isolated plasmid DNA was purified using three methods: RNA in plasmid DNA was precipitated by lithium chloride, RNA was degraded by immobilized RNase A and plasmid DNA was purified using two-phase aqueous system.
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Growth of Escherichia coli on nanocrystalline diamond
Jurková, Blanka ; Kozak, Halyna ; Artemenko, Anna ; Ukraintsev, Egor ; Beranová, J. ; Konopásek, I. ; Kromka, Alexander
In this contribution, we compared the attachment of gram-negative model bacterium Escherichia coli to\nuncoated glass and glass coated by hydrogenated and oxidized NCD films. For attachment experiments,\ncontinuous cultivation in commercially available CDC Bioreactor was used. Antibacterial tests indicated\nhigher attachment of gram-negative model bacterium Escherichia coli to NCD surface compared to uncoated\nglass. We assign this effect to higher roughness of NCD surface compared to glass. Bacterial cells preferred\nthe hydrophobic surface of hydrogenated NCD surface to hydrophilic oxidized NCD for their attachment.
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Semiquantitative bacteriological examination of urine
VITANOVSKÁ, Alena
Urine is a liquid product of metabolism excreted through the kidneys. It refers to the overall health of the body. Urinary tract infections are the second most common disease in the population caused by various pathogens. E. coli is the most frequent pathogen. The increase in resistance of bacterial pathogens to antibiotics is related to the high incidence of persistent infections and their treatment. The main objectives of this thesis are to introduce the bacteriological examination of urinary tract infections and mastering the art semiquantitative bacteriological examination of urine and other processes leading to the diagnosis of urinary pathogens using in the Medical Microbiology Department of Klatovská nemocnice, a.s.. Then, based on the results taken from LIS OLM to evaluate the obtained results and to compare them with the literature. The first part deals with the definition of basic concepts that are associated with urinary tract infections, and its infections and occurring pathogens. The various phases of laboratory procedure are described. In particular, analytical part of laboratory procedure which deals with theoretical description of methods of laboratory diagnostics. The procedures of identification methods, which are used in the Medical Microbiology Department of Klatovská nemocnice, a.s. are described in the methodology. That means semiquantitative examination, microscopy, various biochemical tests for pathogens identification and process for the determination of sensitivity. The research results are evaluated using simple statistics in tables and graphs. In the year 2014 8623 urine samples were examined. 6267 samples came from hospital patients. The samples from hospital patients were for the statistical evaluation. From these samples 1260 were cases of E. coli, Enterococcus 829 cases and 297 cases of Proteus. For further statistical evaluation is carried out with E. coli which determine sensitivity to nitrofurantoin such were 1030. From that 819 women and 211 men. The highest incidence of this pathogen was among women in the age group over 61 years, it were the 542 patients. For men the capture also the highest over the age of 61 years, with 166 patients. The lowest detection of E. coli in both men and women was the lowest compared to other categories of age 7-17.
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The analysis of selected parameters of water quality
KŘÍŽOVÁ, Iveta
Cílem této diplomové práce je provést analýzu vybraných ukazatelů kvality pitné vody a posouzení faktorů, které ji ovlivňují. V první části jsou zachyceny vybrané mikrobiologické ukazatele, tak jak je popisuje literatura, včetně platných norem a vyhlášek. Druhá část popisuje průběh provádění vlastní analýzy, z poskytnutých materiálů akreditovanou laboratoří společnosti AGRO-LA, spol. s r. o., až po vyhodnocení získaných výsledků.
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Development of antibiotic resistance in the region Písek in 2000-2010
MAREŠOVÁ, Lucie
This bachleor´ s dissertation is aimed to the developement of antibiotical resistance by choosed kinds of bacteria, particulary Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae in the region of Písek and also in the whole Czech Republic during the period 2000-2010. There is described matter of the antibiotical resistance, antimicrobial agents and also information about the EARS-Net system in the beginning of this work. The practical part contains the evaluation of resistance spread in the region of Pisek according to the data of clinical microbiology of Nemocnice Pisek, the spread of resistance in the Czech Republic according to the EARS-Net and also the consumption of individual groups of antibiotics according to the State Institute for Drug Control. Antibiotical resistance is spreading constantly, for example because of inappropriate usage of the antibiotics or insufficient information about pathogens that cause infectious diseases the most frequently.
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The Quality of Drinking Water in Public Distribution Systems
SOMPEKOVÁ, Zuzana
This research project was aimed at monitoring the quality of drinking water that is supplied to the inhabitants of small villages. The quality of drinking water produced by small waterworks in South Bohemia, in municipalities Mazelov, Ortvínovice, Doubravka and Rábín, was studied. Sanitary analyses of drinking water samples carried out by the waterworks operators in 2004-2009 showed some variability in the concentrations of free chlorine, nitrates, pH, turbidity and the content of Escherichia coli in all the waterworks during the investigated period. The hypothesis assuming that the quality of drinking water produced by water treatment from small water sources is stable and that it does not vary in some key indicators, such as nitrates, the contents of Escherichia coli etc., throughout the year was not confirmed. The other hypothesis assuming that the number of small water sources used for public drinking water supplies decreases during the period was confirmed. The causes of these changes depend on many factors, such as the location and source of drinking water, the type of treatment plant, and, last but not least, the quality of service and economic potential of the waterworks operators play a negative role.
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