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Porovnání výkonnosti potomstva belgického ovčáka malinois po domácích a zahraničních plemenících
KAMENÍK, Zdeněk
This graduation thesis deals with comparison of performance of offspring's of belgian shepherd malinois - the national and foreign stud dogs. In literary overview describes the history and a present form of the International Examination Rules IPO, breeding Belgian shepherds in the Czech Republic and in the world. It describes complete characteristic Belgian shepherd like physique, body proportions, life requirements. The thesis subsequently pursues impacts affecting dog´s sport performance, breeding methods and breeding dogs and observes me the most common health issues in Belgian shepherd breeding. In the own thesis are analyses of performance of offspring's of national and foreign stud dogs in the position of a father or father´s father (grandfather) in championship ČR BO, championship Czech Republic IPO. There were monitored data on numbers of individuals in specified competitions in monitored years 2012 - 2018. Dogs placed on 1st to 10th positions at MČR IPO and at MČR BO were futher evaluated. By detailed observation and analysis was discovered that for sports cynology are used dogs from foreing breeding descent whether it is after father or dog on the grandfather position. It was found that dogs, which placed to the tenth placed at MČR IPO had been 64% of foreign and 36 % of Czech descent. At championship ČR BO stud dogs of placed placed dogs in ratio 65% foreign and 35 % domestic individuals. The best stud dogs from abroad were Da´Eder vom der Brunnenstad a Solero des Teutones. Jaguar de Alphaville Bohemia belonged to the best national stud dogs.
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The role of F420-dependent oxidoreductases in actinobacteria
Kekrt, Lukáš ; Kameník, Zdeněk (advisor) ; Palyzová, Andrea (referee)
Deazaflavin F420 is an unusual cofactor involved in oxidoreduction reactions in the cells of some microorganisms. The role of F420/F420H2-dependent oxidoreductases has been extensively described in the case of central archaeal metabolism, particularly those of methanogens. In contrast, our knowledge of these enzymes in actinobacteria is limited. This work focuses on the characterization of selected actinobacterial oxidoreductases from luciferase-like hydride transferase family, which putatively use the F420 cofactor. Specifically, Apd6 biosynthetic proteins and their sequence homologs were studied. Three recombinant proteins were prepared and purified and their enzymatic activity was tested in the presence of F420 and a set of putative substrates by means of in vitro reactions. The reaction products were monitored by liquid chromatography with UV and mass spectrometry detection. Some of the expected reaction products were detected in in vitro reactions, confirming that the proteins were catalytically active. Furthermore, one of the proteins surprisingly exhibited an unusual reaction specificity. Key words: Specialized metabolism, actinobacteria, F420 cofactor, redox reactions, reaction specificity, 4-alkyl-L-proline derivates.
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Bioinformatics Tool for the Analysis of Gene Neighbourhoods
Hřebíček, Ondřej ; Kameník, Zdeněk (advisor) ; Větrovský, Tomáš (referee)
The discovery of novel biosynthetic gene clusters (BGCs), particularly in Actinobacteria, is key for the development of new antimicrobials and other bioactive compounds derived from bacterial specialized metabolites. BGC database-independent BGC-finding software represents a largely-unexplored avenue of research capable of finding BGCs missed by mainstream software due to its dependence on databases of published BGCs. In this work, a bioinformatics tool is presented, which seeks to unite the fragmented functions of database-independent BGC-finding software into a single comprehensive package and make it accessible to non-expert users through an easy-to-use graphical user interface. The software utilizes the output of NCBI BLAST containing user-selected marker proteins to identify points of co-localization, and then displays the results through a fully- interactive graphical interface which allows users to further analyse the data. The software's capabilities and limitations have been tested on L-tyrosine-derived and L-leucine-derived 4-alkyl-L- proline biosynthetic gene subclusters. The software was able to find 120 out of 121 manually-found biosynthetic gene subclusters, showcasing the reliability of the core algorithm. In addition, a further 131 putative L-leucine-derived 4-alkyl-L-proline...
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Liquid Chromatography Methods for Analysis of Actinomycete Secondary Metabolites - Potential Antibiotics
Kameník, Zdeněk ; Nesměrák, Karel (advisor) ; Cvačka, Josef (referee) ; Bosáková, Zuzana (referee)
(EN) This dissertation thesis contains scientific results achieved in the field of analytical chemistry, particularly liquid chromatography. The major part of the results has been published in prestigious international journals in five papers. In addition to that, relevant yet unpublished results have been included as well. In general terms, the work presented here contributed to the concerted efforts to tackle the current lack of novel antibiotics. Specifically, high-performance liquid chromatography (HPLC) and ultra high-performance liquid chromatography (UHPLC) techniques coupled to a variety of detection systems have been employed for analysis of antibiotics and actinomycete secondary metabolites. The first thematic part describes the development of liquid chromatography methods for analysis of lincomycin precursors, lincomycin precursor analogues, and lincomycin derivatives. The methods have been applied to study lincomycin biosynthetic pathway and obtain improved lincomycin derivatives by mutasynthesis. The second thematic part aims at investigating alternative approaches for analysis of antibiotics. Firstly, the core-shell particle and the sub-2 μm particle chromatographic columns were compared. The core-shell particle columns compatible with HPLC proved to be a convenient alternative to the...
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Development of miniaturized extraction method used for GC-NCD screening of non-volatile nitroso compounds in malt
Malečková, Michaela ; Sobotníková, Jana (advisor) ; Kameník, Zdeněk (referee)
The aim of this diploma thesis was to develop a miniaturized extraction method for a fast screening of non-volatile nitroso compounds using gas chromatography with a nitroso specific chemiluminescence detection. According to a final methodology, the samples were prepared by extraction of grinded malt using a mixture of pyridine and acetonitrile in ratio 60:40 (v/v). To enhance volatility of the determined analytes, the two-step derivatization using hexamethyldisalazane and N,O-bis(trimethyl)-trifluoroacetamide was used. The total volume of the sample was 200 l and the preparation time after optimization was in total 80 min. The extraction method was connected to a classification method, which can divide chromatographic peaks into the groups of N-nitroso and C-nitroso compounds, and interfering substances. After application of the methods mentioned above to real malt samples, the specific chromatographic peaks of C-nitroso and N-nitroso compounds were selected. Description of their properties and structure suggestion will be a subject of the following study. Keywords Nitroso compounds, malt, extraction, derivatization, gas chromatography, chemiluminescence detector
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Interactions between Schistosoma spp. and their hosts at the metabolome level
Kurečka, Martin ; Kameník, Zdeněk (advisor) ; Mikeš, Libor (referee)
The blood flukes of the genus Schistosoma are important parasites that cause serious chronic diseases in mammals, including humans, in tropical and subtropical countries. Treatment of these diseases is challenging; therefore, new molecular targets are still being sought for the development of vaccines and more effective drugs. To achieve this, better understanding of interactions between the parasite and the host at the molecular level is an important prerequisite. These processes can be studied by quantitative and qualitative determination of metabolite differences in healthy and infected individuals using metabolomics. The work represents a review of low molecular weight substances in tissues and body fluids of schistosome hosts, in which a change in concentration of metabolites putatively related to the infection was observed. Only metabolites with a hypothetical or known mechanism of these changes in the context of infection are covered. The thesis also includes a brief overview of basic methods of analytical chemistry, which are used in studies based on metabolomics. Key words: Schistosoma spp.; intermediate host; definitive host; spectrometry; metabolomics; low molecular compounds
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Biologically active compounds of selected model trematodes
Kurečka, Martin ; Kameník, Zdeněk (advisor) ; Mikeš, Libor (referee)
Trematoda are parasites known for their ability to manipulate their host for survival and reproduction. They have complex life cycles with the intermediate host represented by mollusks and the definitive host, represented by vertebrates. This work focuses on three medically important genera of trematodes: Schistosoma, Fasciola and Opisthorchis. The aim of this work is to summarize biologically active low molecular weight substances that parasites modulate or produce in order to manipulate their host. The result of the work is a literature research of a comparative change in the concentration of metabolites of infected and uninfected trematode hosts with a focus on the analytical method used. Metabolomics deals with a comprehensive analysis of the metabolism of biological samples. It uses spectrometric analytical methods such as nuclear magnetic resonance and mass spectrometry combined with gas or liquid chromatography. Part of the work is also a summary of the importance, development and perspectives of metabolic profiling in parasitology. Current research in this area focuses mainly on vertebrate hosts. In addition, for vertebrate hosts, the sum of studied substances is still much broader than that in intermediate hosts. In intermediate host studies also focus on different types of substances,...
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Ergothioneine and mycothiol in the biosynthesis of lincosamides
Seidlová, Bára ; Kameník, Zdeněk (advisor) ; Kopecký, Jan (referee)
Specialized microbial metabolites are described as low-molecular-weight bioactive compounds, which are dispensable for the growth, evolution, or reproduction of its producer. This group of substances includes the lincosamides, which are produced mainly by the bacteria of the Streptomyces genera. Apart from other precursors, two low-molecular-weight thiols, ergothioneine and mycothiol, are essential participants of the lincosamide biosynthesis. Mycothiol (MSH) serves in this pathway as a source of sulphur, on the other hand, ergothioneine (ESH) constitutes a conjugate with the aminosugar moiety of lincosamide structure. The conjugate is condensed with an activated amino acid, which is catalyzed by an unusual enzyme to form a core of the lincosamide molecule. The objective of this diploma thesis is to isolate the conjugate of ESH and aminooctose, which serves as a substrate of the LmbD biosynthetic protein. Another aim is to study the links between the thiol metabolism and the biosynthesis of three lincosamides, lincomycin, celesticetin, and intervencin, which are produced by different bacterial strains. Bacterial strains were cultivated under laboratory conditions and methods of liquid chromatography with UV and MS detection were used for the analysis. The parameters of the methods were developed...
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Development of miniaturized extraction method used for GC-NCD screening of non-volatile nitroso compounds in malt
Malečková, Michaela ; Sobotníková, Jana (advisor) ; Kameník, Zdeněk (referee)
The aim of this diploma thesis was to develop a miniaturized extraction method for a fast screening of non-volatile nitroso compounds using gas chromatography with a nitroso specific chemiluminescence detection. According to a final methodology, the samples were prepared by extraction of grinded malt using a mixture of pyridine and acetonitrile in ratio 60:40 (v/v). To enhance volatility of the determined analytes, the two-step derivatization using hexamethyldisalazane and N,O-bis(trimethyl)-trifluoroacetamide was used. The total volume of the sample was 200 l and the preparation time after optimization was in total 80 min. The extraction method was connected to a classification method, which can divide chromatographic peaks into the groups of N-nitroso and C-nitroso compounds, and interfering substances. After application of the methods mentioned above to real malt samples, the specific chromatographic peaks of C-nitroso and N-nitroso compounds were selected. Description of their properties and structure suggestion will be a subject of the following study. Keywords Nitroso compounds, malt, extraction, derivatization, gas chromatography, chemiluminescence detector
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