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PROBIOTIC GENES OF SIGNIFICANT LACTIC ACID BACTERIA IN FOOD
Konečná, Jana ; Ševčovičová,, Andrea (referee) ; Doškař, Jiří (referee) ; Španová, Alena (advisor)
Isolation of deoxyribonucleic acid (DNA) is an important step in the molecular diagnostics of microorganisms. A high quality of isolated DNA is necessary for DNA amplification by the polymerase chain reaction (PCR). The conventional DNA isolation using phenol chloroform extraction and DNA precipitation in ethanol is time consuming and requires the use of toxic phenol. Magnetic separation techniques using magnetic solid particles are one of modern methods to speed up the nucleic acids isolation. The aim of this work was to use two different types of magnetic particles for solidphase DNA extraction. The amounts of DNA in separation mixtures were measured using ultraviolet spectrophotometry (UV). The first experimental conditions were tested on chicken erythrocytes DNA. Phosphate buffer (pH 7, 7.6 and 8) was used for adsorption of DNA on magnetic particles. It was shown that approximately almost one half of DNA was adsorbed to the particles. The elution conditions of DNA were also optimized. Secondly, bacterial DNA was tested. This DNA eluted from the particles was in PCR ready quality. High resolution melting (HRM) curve analysis is a simple, low-cost method for amplicon discrimination and easy connection with real-time polymerase chain reaction (PCR). In this contribution, we report rapid species identification of strains belonging to the Lactobacillus group using HRM-PCR. Three different DNA isolation methods were used in this work: phenol extraction, separation using magnetic particles and commercial kit. Ten sets of targeted gene fragments primers (LAC1 – LAC2, LAC2 – LAC4, P1V1 – P2V1, Gro F – Gro R, 3BA-338f – Primer 1, V1F – V1R, CHAU - V3F – CHAU - V3R, CHAU - V6F – CHAU - V6R, poxcDNAFw – poxPromRVC, poxcDNAFw – poxPromRVT) were tested for amplification of the 16S rRNA gene. Use of GroF/R and LAC2/4 primers pairs successfully identify strains belong to the Lactobacillus group. The variance between used extraction methods for evidence of HRM curves was found.
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Magteic particles and their applications in biotechnology
Knápková, Monika ; Konečná, Jana (referee) ; Trachtová, Štěpánka (advisor)
The thesis is focused on the magnetic particles which are used in several biotechnological applications. The theoretical part deals with the specific properties of these nanoparticles and materials of which the nanoparticles can be made. There are also mentioned some of the biotechnological applications of magnetic particles. During the experimental part, selected types of magnetic particles were used to isolate nucleic acid. The quality of the isolated DNA with respect to purity was evaluated using the polymerase chain reaction and its modifications. High resolution analysis (HRM analysis) was also used to verify the quality of the isolated DNA and to resolution Lactobacillus casei and Lactobacillus rhamnosus. DNA isolation using magnetic carriers was successful. Commercially available MPG microcarriers and magnetic microparticles Fkol 77ox were the most suitable. In terms of purity magnetic nanoparticles F79/L3-PLL were the most suitable for the DNA isolation. The resolution of bacterial strains of Lactobacillus casei and Lactobacillus rhamnosus was not successful.
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Nurses' duty of conifidentiality and the consequences of its breach
Benešová, Pavla ; Prošková, Eva (advisor) ; Konečná, Jana (referee)
The aim of my thesis was to find out what is the legal awareness of nurses in the issue of confidentiality. Furthermore, it was examined whether nurses know what are the consequences for unauthorized access to the medical records. The theoretical part of this thesis describes the mandatory confidentiality issue and its consequences. And it defines the concepts of personal and sensitive personal data. In addition the conditions under which it is required to break confidentiality and the consequences of breach of legal obligation are mentioned. Describe the legal responsibility of nurses. In the practical part of the work I use qualitative methods of data collection, interview, for me to respond to major and minor research questions. I use interviews conducted with nurses, for doing analysis of the case and to describe the measures taken by the employer at the Municipal Hospital in Litoměřice.
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Sacred Choir Music by Vít Clar
Konečná, Jolana ; Slavický, Tomáš (advisor) ; Douša, Eduard (referee)
- 48 - 9. RESUME Bakalářská práce je zaměřena na život a dílo Víta Clara. V práci byly použity ukázky ze sborové duchovní tvorby, byly shrnuty kompoziční prostředky. V závěru práce je zhodnocení vybraných skladeb z hlediska interpretačního. Pozornost byla věnována významným českým osobnostem současnosti v české hudební pedagogice posledních desetiletí. Nemalou překážkou byl stav pramenů a nedostatek dohledatelných materiálů, stejně tak i čekání na zpracování notové pozůstalosti. Naopak byly potěšitelné postřehy a odpovědi kolegů a Clarových studentů, zejména jejich milé vzpomínky na nezapomenutelného skladatele a pedagoga. Tato práce je věnována památce Víta Clara im memoriam. 9. RESUME This thesis focuses on the life and work of Vít Clar. It presents some examples of sacred music for choir and summarizes Clar's musical composition elements. The last chapter evaluates selected compositions from the interpretation point of view. This text also pays attention to significant Czech personalities in contemporary musical education of several past decades. Work on this thesis was considerably hampered by the poor conditions of resources and the lack of available materials as well as by the duration of Clar's score estate's processing. On the other hand, observations and answers of Clar's colleagues and students,...
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Non-traditional foods containing probiotic bacteria
Sobotková, Markéta ; Konečná, Jana (referee) ; Trachtová, Štěpánka (advisor)
Probiotic microorganisms are contained in common dairy products as well in unconventional probiotic products consumed especially in Asian countries. In the thesis is shown overview of chosen less common foodstuffs containing probiotic microorganisms, which no belong to fermented dairy products. In the thesis are discussed positive health benefits of probiotics foodstuffs on the host. The experimental part deals with the identification of probiotic microorganisms, which are contained in chosen probiotic product from Asia. DNA was isolated from the product by use phenol extraction and using magnetic particles. Probiotic microorganisms in chosen product were detected using method real-time polymerase chain reaction (qPCR).
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Identification of probiotic bacteria in complex food matrices and dietary supplements
Cesnak, Filip ; Konečná, Jana (referee) ; Trachtová, Štěpánka (advisor)
Theoretical part of the thesis deals with the general characteristics of probiotic bacteria, their effect on human health and with technological demands imposed on them. Furthermore the thesis deals with actual methods of bacterial identification, especially on the base of polymerase chain reaction. Presence of the declared probiotic bacteria was analysed with the polymerase chain reaction (PCR) and the DNA was isolated from the sample of food additive with the use of magnetic particles.
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Assessment of Market Value of the Easement on the Route of the Sewage Pipelines
Konečná, Jana ; Hlavinková, Vítězslava (referee) ; Hrubanová, Michaela (advisor)
This dissertation deals wtih a problematice of easements of real estates in Czech Republic.The main goal of the dissertation is to estimate the value of considered easement and value of the family house burdened with the easements, consider and give reasons for possible devaluation of the real estate by making the easements. In the theoretical part are defined the terms connected with the real estate business, methods of valuation of the real estates and terms from land registry. Legislation of easements, methods and proceduresof valuation of easementsand valuationof real estates with easements follows. Practical part is about valuation of easements. Municipakl authority in Vyškov reallized construction of sewers in local parts Lhota and Rychtářov. The sewers goes through lands thats why the easements are established for the land owners. The easements are provided with conrtracts. The final part of the dissertation includes the summary and comparison of easements in 2011 and 2014, example of valuation of family house in local part Rychtářov and valuation of the influence of easement on the real estate.
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