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Reeducation of mechanical knowledge in mathematics during private supplementary tutoring
Novotná, Gabriela ; Vondrová, Naďa (advisor) ; Novotná, Jarmila (referee)
The diploma thesis is aimed at reeducation of mechanical knowledge in the area of fractions. Since every student has different needs and pace in his/her process of gaining knowledge, the reeducation is set into their private supplementary tutoring lessons. In the theoretical part, the basic concepts (how is the private supplementary tutoring understood in this thesis, which students are taken into consideration, and what is mechanical knowledge) are explained based on Czech and foreign literary sources, subsequently, the constructivist way of teaching, which is suitable for the reeducation of mechanical knowledge, is described. The second part of this thesis is constituted by the practical implementation of the theoretical findings into three case studies. Three students were diagnosed with mechanical knowledge in the area of fractions. Subsequently, their knowledge was being reeducated during their private supplementary tutoring lessons. The process of reeducation of each student is described in detail, moreover, a kind of manual describing what should not be avoided when dealing with this topic is included. KEY WORDS mechanical knowledge, reeducation, private supplementary tutoring, fractions 5
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Pronounciation and perceptual acceptability of the sound r in Czech speeches by foreign speakers whose L1 is German
Novotná, Gabriela ; Veroňková, Jitka (advisor) ; Martínek, František (referee)
The topic of this diploma thesis belongs to the field of language acquisition and language learning. It aims to explore Czech as a second and foreign language of speakers whose mother tongue is German, namely concerning the pronunciation of the r-sound and its perceptual acceptability for Czech native speakers. The theoretical part summarizes the area of pronunciation learning and acquisition in a second and foreign language and outlines various pronunciation difficulties German speakers tend to have in Czech. It deals with the rhotics in general and subsequently focuses on the r- sound, both in Czech and in German, as well as in mutual comparison. In the last chapter, the combinatorial qualities of the r-sound which are used as a basis for the practical part are discussed. The practical part describes the conducted research. Five recordings of German students were collected. The students were staying in Prague for one semester with the Erasmus programme exchange and were learning Czech for the first time. For the recordings a written text was constructed which the students read individually. The text emphasized words including the r-sound. From the recordings, various key words were cut off and a perceptual test was compiled and given to Czech native speakers to evaluate the pronunciation of the...
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Critical sites determining the resistance phenotype of ABC proteins from the ARE subfamily and the molecular mechanism of their function
Lenart, Jakub ; Balíková Novotná, Gabriela (advisor) ; Melter, Oto (referee) ; Branny, Pavel (referee)
Vga(A) and Msr(A) are resistance proteins belonging to the ARE subfamily of ABC -F proteins. They confer resistance to inhibitors of the peptidyltransferase center. It has been proposed that the mechanism of resistance is based on interaction with a transmembrane partner that forms the functional transporter. Their ribosomal function has been described by cryoelectron microscopy of ribosome complexes with ABCF mutants unable to hydrolyze ATP. However, the exact mechanism of resistance is not yet known. We have produced the mutant proteins combining the four amino acid residues in Vga(A) and Vga(A)LC at the linker tip, and we were the first to describe the effects of substrate specificity of the single mutants. Amino acid positions 212 and 220 are important for resistance to lincosamides and pleuromutilins, respectively, while position 219 is responsible for resistance to streptogramin A. Each amino acid property plays a critical role in conferring antibiotic specificity, as confirmed by the fact that amino acid substitution at position K218T in the Vga(A) protein causes the shift in resistance from streptogramins to lincosamides and pleuromutilins. The mechanism of resistance conferred by Vga(A) is ribosomal protection. This is supported by the fact that the rate of [3H]-lincomycin accumulation in...
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Mechanism of inducible gene expression of resistance protein Vga(A)LC from Staphylococcus haemolyticus.
Novotná, Michaela ; Balíková Novotná, Gabriela (advisor) ; Lišková, Petra (referee)
The staphylococcal protein VgaA belongs to ARE ABCF family, which confers resistance to ribosome binding antibiotics by the target protection mechanism. VgaA confers resistance to lincosamides, streptogramins A and pleuromutilins and thus provides the so-called LSAP resistance phenotype. The expression of resistance genes often reduces fitness in the absence of an antibiotic, therefore the expression of resistance genes is often tightly controlled and triggered only in response to the presence of an antibiotic to which the protein confers resistance. The inducible expression has also been observed for the vgaA gene, nevertheless, its mechanism has not been elucidated. In the diploma thesis, it was shown that the vgaALC gene from Staphylococcus haemolyticus is regulated by ribosome-mediated attenuation. The mechanism is based on the detection of translation inhibitors via a ribosome translating a special regulatory open reading frame (uORF), which is part of an attenuator located in the 5' untranslated region of the mRNA. The vgaALC gene is regulated at the transcriptional level in response to LSAP antibiotics. Antibiotic specificity of induction is affected not only by the nature of the peptide encoded by uORF but also by the antibiotic specificity of the resistance protein. Fluorescence microscopy...
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Enterococcus spp. as a reservoir of resistance genes.
Zajíčková, Adéla ; Lichá, Irena (advisor) ; Balíková Novotná, Gabriela (referee)
Bacteria of the genus Enterococcus are categorised among common nosocomial pathogens. They are a significant reservoir of resistance genes to a majority of antibiotics and exhibit an intrinsic resistance to low levels of beta-lactams, glycopeptides, aminoglycosides, streptogramines and lincosamides. The aim of this paper is to review the main resistance genes and other mechanisms involved in the resistance of bacteria of this genus to antibiotics. The paper is mainly focused on the resistance to beta-lactam antibiotics, which is provided by the expression and mutations of low-affinity PBPs, the individual van resistance types mediating resistance to vancomycin, and the expression of enzymes capable of modifying the functional groups of aminoglycoside antibiotics. The paper also describes the resistance to newer antibiotics that are used to treat vancomycin-resistant isolates. The resistance to individual antibiotics can arise from the coding of their own chromosomal genes or entire signaling pathways leading to a reduction in the effect of antibiotics, the acquisition of genetic mutations, and especially the spread of new resistance genes by horizontal transfer.
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Reeducation of mechanical knowledge in mathematics during private supplementary tutoring
Novotná, Gabriela
The diploma thesis is aimed at reeducation of mechanical knowledge in the area of fractions. Since every student has different needs and pace in his/her process of gaining knowledge, the reeducation is set into their private supplementary tutoring lessons. In the theoretical part, the basic concepts (how is the private supplementary tutoring understood in this thesis, which students are taken into consideration, and what is mechanical knowledge) are explained based on Czech and foreign literary sources, subsequently, the constructivist way of teaching, which is suitable for the reeducation of mechanical knowledge, is described. The second part of this thesis is constituted by the practical implementation of the theoretical findings into three case studies. Three students were diagnosed with mechanical knowledge in the area of fractions. Subsequently, their knowledge was being reeducated during their private supplementary tutoring lessons. The process of reeducation of each student is described in detail, moreover, a kind of manual describing what should not be avoided when dealing with this topic is included.
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Characterization of the ABC-F protein Sco0636 in Streptomyces coelicolor
Pinďáková, Nikola ; Balíková Novotná, Gabriela (advisor) ; Mikušová, Gabriela (referee)
The main topic of this diploma thesis is ARE (resistance) proteins from the ABC-F family of the second class of ABC proteins. ARE proteins confer resistance to antibiotics that bind to a large ribosomal subunit and therefore inhibit proteosynthesis. One of the ARE proteins is the Lmr (C) protein, which is part of the linkomycin biosynthesis cluster of Streptomyces lincolnensis, and according to new results, Lmr (C) does not have to be just resistant protein but may have also regulatory function. We decided to study Sco0636, the closest homologue to Lmr (C) in Streptomyces coelicolor, which is a model organism in the study of secondary metabolism. Thanks to the production of color pigments, it is possible to monitor the effect of ARE proteins on secondary metabolism directly on the plates. I prepared the deletion mutant and the strain with constitutive expression of sco0636, and observed the effect on the phenotype. I followed the production of a blue asset and set a minimum inhibitory concentration to selected antibiotics, which bind to the ribosome. I have found that Sco0636 gives high resistance to tiamulin and so it has been named TiaA. The deletion of gene sco0636 accelerated production of actinorodine, and constitutive expression of this gene slowed down production. Keywords: ABC proteins,...
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The function of ABCF proteins in bacteria
Mičke, Bianka ; Balíková Novotná, Gabriela (advisor) ; Lišková, Petra (referee)
Translation belongs to the most basic processes which happens in the living cells. It is the last step of proteosynthesis when genetic information encoded by the mRNA is transformed into the protein on a ribosome. Organisms have developed a wide range of mechanisms that can regulate it's needs. I focused on one of them - ABCF proteins. This protein group is a member of the ABC transporters superfamily but they haven't a transmembrane domain and their purpose is protect the ribosomes from antibiotics that bind 50S ribosomal subunit or interact with the ribosomes and influence ribosomal functions. Today, we can divide ABCF proteins into the two functional groups: antibiotic resistence proteins (ARE) and proteins with the regulatory functions. The translational regulatory function has been confirmed There is 45 ABCF protein subfamilies spread through the bacteries and eukaryotes but many essential informations like the structure and exact function of them are still missing. My bachelor thesis is analysis and summary of facts that are known about the bacterial ABCF proteins. Key words: ABCF proteins, antibiotic resistence, ARE, translational regulation, ribosome, translation, translational factors
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The function of ClpX chaperone in bacteria
Kýr, Jan ; Balíková Novotná, Gabriela (advisor) ; Šiková, Michaela (referee)
Intracellular proteolysis is an essential regulatory process that affects cellular physiology. Since proteolysis destroys proteins irreversibly, this process must be strictly controlled. The AAA+ proteins are the key factors in regulated proteolysis in bacteria. These proteins consist of two functional domains, the AAA+ chaperone domain and the protease domain. One particular group of these AAA+ protein is the Clp protein family. Functional domains of the Clp family are formed by seperate proteins. The hexameric unfoldase ClpX is a member of this protein family. This unfoldase can interact with the highly conserved ClpP protease to form a ClpXP proteolytic complex. This proteolytic complex utilizes the energy of ATP binding and hydrolysis to unfold and translocate the specifically tagged substrate into the ClpP degradation chamber. Substrate recognition is mediated by the binding of ClpX to short unstructured sequences called degradation tags. ClpX recognizes several degradation tags, but the most important one is recognition of the ssrA degradation tag, which is the output of the tmRNA ribosome rescue system. Although ClpX interacts with ClpP, it affects a variety of cellular processes such as the expression of virulence factors or the adaptation to stress factors, ClpX can work independently of...
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