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Analysis of adenosine triphosphate and adenosine diphosphate by HPLC-MS/MS
Černá, Martina ; Coufal, Pavel (advisor) ; Bosáková, Zuzana (referee)
In this bachelor thesis, ADP and ATP samples were analysed and detected with HPLC- MS/MS method. Approximate limit of detection (LOD) for these particular substances were found and their values were compared with the LOD values published in the literature obtained via the same methods and under very similar experimental conditions. Our limits of detection for nucleotides were comparable with the limits described in the literature. Mass spectrometry analysis was performed in the positive and the negative mode of multiple reaction monitoring analysis and electrospray was used for the analyte ionization. The optimal conditions for high performance liquid chromatography of ATP and ADP analysis were acquired on a ZIC - HILIC column with the mobile phase of 75:25 (v/v) acetonitrile / 10 mM ammonium acetate. Ammonium acetate buffer was adjusted to pH of 7.15 and the separation was done under the isocratic elution.
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Chemical enucleation in mammalian oocytes
Černá, Martina ; Krylov, Vladimír (advisor) ; Petr, Jaroslav (referee)
The common techniques of enucleation are inexact and time-consuming. Nuclear DNA staining with Hoechst is frequently used, whereas the staining agent is toxic for the oocyte and has negative impact on its further development. Chemical enucleation is a method which facilitates the production of cytoplasts in higher amount and shorter time. Chemical enucleation has a potentional use in the field of biotechnology, especially in nuclear transfer methods (therapeutical, reproductive cloning). The chemical enucleation was successfully used on several model animals: mouse, sheep, cattle. The technique is beeing continually improved on pigs, regarding their similarity to human where it should be consequently applied. It was discovered that same chemicals have different effect on oocytes of each species and therefore it is necessary to examine each species separately. In our study we use porcine oocytes for chemical enucleation. We determined demekolcine in concentration 0,4 Ug/ml and acting time 30 minutes as the most suitable for chemical enucleation. Furthermore we used cytochalasin B in concetration 7,5 Ug/ml and acting time 10 minutes. The protrusion rate correlates with quality of oocytes. In other experiment we focused on distribution of mitochondria after fusion of normal and enucleation oocyte. We found...
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Ways of application of drama methods in german language teaching
Černá, Martina ; Nečasová, Pavla (advisor) ; Švermová, Dagmar (referee)
TITLE: Ways of application of drama methods in German language teaching ABSTRACT This diploma thesis deals with ways of application of drama methods in teaching the German language. The thesis is divided into two parts. The first part describes the theoretical background of educational drama and communicative language teaching. Also, the relation of the Framework Educational Programme for primary education to drama and foreign language teaching is characterized. The main part of this thesis contains a suggestion for integrating drama methods into teaching the German language and a selection of drama-oriented exercises. Further, the process and the results of my research, the main purpose of which was to examine the influence of drama methods on communicative competence in pupils, are described here. KEY WORDS: Educational drama, communicative competence, foreign language teaching
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Determination of adenosine triphosphate and adenosine diphosphate in real samples
Černá, Martina ; Coufal, Pavel (advisor) ; Zima, Jiří (referee)
The aim of the diploma thesis was to find optimal conditions of high pressure liquid chromatography for the detection and quantification of two common nucleotides, namely adenosine diphosphate and adenosine triphosphate, as well as to perform an analysis of these in real life samples of citrus fruits and plant extracts. Further aim of the project was to determine the limits of detection and quantification of adenosine diphosphate and adenosine triphosphate under the optimized conditions and using these to compare the sensitivity of given detectors. To achieve this HPLC-UV, capillary HPLC-DAD and HPLC-MS apparatus were used. With the help of HPLC with UV detection and capillary HPLC with diode array detector, the calibration curves of the mixture of analytes were measured and the limits of detection as well as quantification of adenosine diphosphate and adenosine triphosphate were determined. Separation of the analytes up to the base line using HPLC-UV and capillary HPLC-DAD was achieved under the conditions of ion pairing chromatography. Column C18 was chosen as an appropriate column. The mobile phase included phosphate buffer, acetonitrile and tetrabutylammonium bisulphate as an ion pairing reagent. The separation was performed with gradient elution. Conditions for analysis using LC-MS were...
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Phonetic features in loud reading of words and non-words, and their perceptual impact
Fischerová, Jana ; Volín, Jan (advisor) ; Černá, Martina (referee)
The recipient in communication is influenced by many complex phenomena through which he or she subconsciously evaluates some personality traits of the speaker. In the current study we are dealing with the question of whether phonetic features also enter into this personal evaluation and how much they can influence perception. As research material we used recordings of 20 speakers 11-15 years old, from which 80 audio items were selected. The items varied by their length (two-syllables vs. four-syllables), existence in the lexicon of the Czech language (real words vs. non-words) and also in the number of speech problems that occurred in them (reduction, assimilation, non-continuous speech etc.). These problematic moments were identified before the creation of the perceptual test and scored by the author of the study and by another student of phonetics. These score points served as the basis for an objective phonetic evaluation, which was later related to the assessment from the respondents. The perceptual test (with versions A and B) was made up of 40 items and was divided into 5 parts according to questions which were concerned with different angles such as the personality traits of diligence and acceptability. These features are part of the Five-Factor Model of Personality and can be included under...
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Educational game activities with chemical theme
Černá, Martina ; Beneš, Pavel (referee)
The rigorous thesis deals with study of one of the method, which go to development of creative genius and activity of students, namely with game. The main goal of the thesis is to explore problems of the game activities, inclusive of conditions, possibilities and advantages of their introduce to teaching chemistry at the second grade of basic school and to present suggestions for their realization. The thesis contains also fifteen concrete game activities with their rules and methodical hints. By means of the guest-size pedagogical research, practised on the choiced sample of the pupils from six basic schools, the prediction, that the result of teaching with games are better than results of teaching, where game activities weren't used, was ratified. Powered by TCPDF (www.tcpdf.org)
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Lexicon evaluation of Children attending Speech Therapy Clinics
Černá, Martina ; Durdilová, Lucie (advisor) ; Hájková, Vanda (referee)
This thesis deals with lexicon evaluation of children attending speech therapy clinics. Compared to other developed countries, there is no modern standardized vocabulary test in the Czech republic. We assume that under the given conditions, procedures and materials used in individual clinics are more or less different. The mail goal of the work lies in the description of the methods used in the selected clinics. In the theoretical chapters we summarize main theoretical knowledge on vocabulary presented in the literature, both Czech and foreign. The focus of the thesis lies in the qualitative research that was conducted in order to identify particular materials used to assess children's vocabulary and describe them in detail. Research results are not limited to the detailed description of these procedures, but also suggest concrete proposals for follow-up work. Such work should focus especially on beginning a process of creating of a standardized diagnostic material allowing more objective quantification of children's skills in the lexical-semantic field. Then clinical speech therapy would gain instrument through which could be better compared not only performance of particular client at various stages of speech therapy, but also different groups of clients between each other, which would clearly be...
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The effect of metabolic surgery in obese patients to calcium-phosphate metabolism and serum concentrations of some micronutrients
Černá, Martina ; Raška, Ivan (advisor) ; Michalská, Dana (referee)
This thesis focuse on bariatric-metabolic surgery and its impact on serum concentrations of various metabolic parameters. The thesis is divided into two parts - theoretical and practical. Basic facts about bariatric-metabolic surgery such as its history, indications, contraindications, surgical methods and their results, nutrition after surgery and the frequency of micronutrient's deficits are described in the theoretical part. Furthermore, information about calcium-phosphate metabolism and metabolites such as calcium, phosphorus, magnesium, vitamin D, vitamin K, parathyroid hormone and osteomarkers is also included. The influence of nutrition and nutritional status on the quality of bone tissue is also mentioned. Last section of the theoretical part is focused on the important vitamins and minerals. The practical part discusses measurements of selected metabolites in serum before and half a year after the surgery. Results of questionnaires which were given to respondents are also included. Questionnaires were focused on lifestyle, nutrition, physical activities and whether the patients take some supplements of micronutrients and protein concentrates. The goal of this work is to evaluate the effect of the surgery on serum concentrations of metabolic parameters and their effect on the health and...
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Chemical signals and reproductive processes of the house mouse (Mus musculus)
Černá, Martina
The aim of my thesis was to identify proteins involved in chemical communication and especially those that are involved in sexual signalling. Volatile chemical signals are transported with lipocalins in their beta-barrel structure to present their ligands to receptors or out of the body. Thus, I focused on the identification of these proteins in saliva and vaginal secretion of the house mouse using proteomic and transcriptomic approaches. Due to a cyclic manner of reproduction and its hormonal control, I have also focused on the role of estradiol on sperm phenotype in the laboratory mouse. We have identified an elevated sexual dimorphism in several lipocalins (i.e. 10 out of 20) in the saliva proteome where they may play a role in sexual signalling (i.e. similar to their described roles in the mouse urine). Interestingly, vaginal secretion also contains lipocalins and they rise from proestrus to estrus and remain steady during metestrus. Such variation provides evidence that they serve sexual signalling, however, due to their elevated levels during metestrus it is most likely that their ligands function as signals and not the proteins themselves. On the level of sperm phenotype, we have provided evidence, that experimental concentrations of estradiol have differential effects on sperm. This is due...
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