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Development of a novel method for nucleotide pool analysis in bacterial cells
Zborníková, Eva ; Rejman, Dominik (advisor) ; Pacáková, Věra (referee) ; Kozlík, Petr (referee)
(EN) This thesis deals with the determination of nucleotides in bacterial cells. Nucleotides play crucial role in most of the metabolic pathway. Determining their concentrations could give us important clues about the influence of internal and external conditions on the bacterial metabolism. Previously published papers dealing with the analysis of nucleotides and other intracellular metabolites can be divided into two groups according to the analytical approach: a) metabolomic approach and b) targeted approach dealing with narrow group of target analytes. In the case a) most authors use the state-of-the-art LC-MS/MS technique, whereas in the case b) robust UV detection coupled mainly to IP-LC is widely used. The aim of this study was to combine both approaches to obtain a method for routine analysis that would take advantages of mass detection, such as sensitivity and selectivity, while avoiding the need of demanding optimization of MS/MS transitions and expert service. The main purpose of the newly developed HILIC-MS method is its universal applicability in most biological and biochemical laboratories.
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Development of a novel method for nucleotide pool analysis in bacterial cells
Zborníková, Eva ; Rejman, Dominik (advisor) ; Pacáková, Věra (referee) ; Kozlík, Petr (referee)
(EN) This thesis deals with the determination of nucleotides in bacterial cells. Nucleotides play crucial role in most of the metabolic pathway. Determining their concentrations could give us important clues about the influence of internal and external conditions on the bacterial metabolism. Previously published papers dealing with the analysis of nucleotides and other intracellular metabolites can be divided into two groups according to the analytical approach: a) metabolomic approach and b) targeted approach dealing with narrow group of target analytes. In the case a) most authors use the state-of-the-art LC-MS/MS technique, whereas in the case b) robust UV detection coupled mainly to IP-LC is widely used. The aim of this study was to combine both approaches to obtain a method for routine analysis that would take advantages of mass detection, such as sensitivity and selectivity, while avoiding the need of demanding optimization of MS/MS transitions and expert service. The main purpose of the newly developed HILIC-MS method is its universal applicability in most biological and biochemical laboratories.
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Phosphanucleosides containing 1-hydroxymethylphospholane 1-oxide ring
Páv, Ondřej ; Zborníková, Eva ; Buděšínský, Miloš ; Rosenberg, Ivan
Novel phosphanucleosides containing 1-hydroxymethylphospholane 1-oxide ring were synthesized as compounds with potential biological activity. A ring-closing metathesis was employed to prepare a phospholene precursor, which was then converted into either 4-azido-3-hydroxy-1-hydroxymethylphospholane 1-oxide or 3-azido-1- hydroxymethylphospholane 1-oxide derivatives. Subsequently, nucleobase was introduced using a nucleobase construction. The biological activity of prepared compounds was evaluated.
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Insights into the mechanism of action of bactericidal lipophosphonoxins
Panova, Natalya ; Zborníková, Eva ; Šimák, Ondřej ; Krásný, Libor ; Kolář, M. ; Látal, T. ; Seydlová, G. ; Rejman, Dominik
The advantages offered by antibiotics in the treatment of infectious diseases are endangered due to the increase in the number of antibiotic-resistant bacterial strains. This reduces the efficiency of antibiotic treatments and poses a serious health and economical problem. Currently, the need for novel antibiotics is becoming increasingly apparent. Recently, we discovered a series of compounds termed lipophosphonoxins exhibiting selective cytotoxicity towards gram-positive bacterial cells. The attempt to elucidate the mode of action of lipophosphonoxins is presented here.
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