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Microbiological excursions
Záhořová, Karolína ; Pavlasová, Lenka (advisor) ; Skýbová, Jana (referee)
This thesis work is aimed at the meaning of excursion in teaching biology at secondary vocational school for thematic complex of microbiology. The topic of microbiology is not attractive for students of secondary vocational school and it is very important to find out what the impact of the excursion will be on the knowledge of students who will participate on the excursion Part of this work is evaluation of excursion as an alternative method of teaching and its overall importance in the teaching of biology. Diploma thesis work brings an overview how the individual stages of the excursion should theoretically look like. It provides list of institutions and factories which may be asked for microbiology excursion in practice. Next part includes completely described excursion in brewery U Fleků including preparation realization and evaluation. Excursion was taken by two classes of different fields with identical pre concept of their studies. Part of the school phase of the excursion is testing students in the form of pre test with the knowledge of microbiology theme, particularly beer production and yeasting process. It is followed by excursion part of brewery and at the end comes diagnostic school part when the students will be again tested in the form of post test. At the end of this work there are...
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Study of membrane transport processes in yeast using potentiometric fluorescent porbe diS-C3(3)
Bartl, Tomáš ; Gášková, Dana (advisor) ; Krůšek, Jan (referee)
1 Title: Study of membrane transport processes in yeast using potentiometric fluorescent probe diS-C3(3) Author: Tomáš Bartl Department: Institute of Physics of Charles University Supervisor: doc. RNDr. Dana Gášková, CSc., Institute of Physics of Charles University Abstract: Yeast membranes contain a number of transporters. Some are responsible for flow of nutrients to the inside of the cell, others for disposing of waste and foreign substances and some for transport of small ions or protons across the membrane. The focus of this work is on the activity of specific transport membrane proteins, so-called MDR pumps, which are responsible for transport of foreign substances or drugs, out of the cell. Using the series of mutant strains of the yeast Saccharomyces cerevisiae (AD1-3, AD1-8 and AD12) differentiated in the presence of specific MDR pumps in their membrane, an influence of various chemical substances on the intracellular concentration of the potentiometric fluorescent probe diS-C3(3), which is actively being transported out of the cell by some of the MDR pumps, was observed. By the examination of the effect of 2-deoxyglucose we proved the active contribution of not only the main MDR pump, Pdr5p, but also of some other pumps, in lowering the intracellular probe concentration. It was observed that...
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TORC1 pathway regulators in yeast cells
Trubitsyna, Yana ; Palková, Zdena (advisor) ; Mašek, Tomáš (referee)
TORC1 ("Target Of Rapamycin") is highly conserved serine-threonine kinase that regulates cell growth, metabolism, proliferation and apoptosis. Some of the regulators and mechanisms of regulation of TORC1 activity are conserved in different eukaryotic organisms. In mammalian cells mTORC1 ("mammalian Target Of Rapamycin") is present that is homologous to yeast TORC1. The correct regulation of TORC1 activity is required for vitality of organisms and thus TORC1 and its regulations are investigated by many researches. A mutation in a single protein that is a part of TORC1 regulating complexes may result in a serious damage to the cell and to the organism. This thesis aims to describe the known TORC1 regulators in yeast and, in some cases, to compare TORC1 and mTORC1 regulations. Information on mTORC1 regulations that have not yet been identified in budding yeasts is also included. Keywords - TORC1, mTORC1, regulation, activity, yeast
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Yeast as the model for for neurodegenerative diseases
Motyčková, Věra ; Schierová, Michaela (advisor) ; Zikánová, Blanka (referee)
Yeasts are traditional model organisms, which can be, surprisingly, used for studying neurodegenerative diseases such as Parkinson, Alzheimer and Huntington diseases. These diseases affect mostly elder population. It's because of decrease of cell control. Common denominator of these diseases are impaired vesicular transport, protein synthesis and degradation, increasing oxidative stress and ROS. The main cause of neurodegenerative diseases is protein misfolding, which provokes cytotoxic protein aggregation. Modified yeast strains can be used to identify genes and proteins that affect cytotoxicity of these key proteins. If these genes have human orthologues, they could be therapeutic targets. The aim of this work is to summarize methods that study causes of neurodegeneration in yeast and to describe the main results discovered in yeast models. Key words: yeast, model system, Parkinson disease, Alzheimer disease, Huntington disease
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Protein synthesis in cellular stress
Cienciala, Martin ; Pospíšek, Martin (advisor) ; Holá, Dana (referee)
Environmental stress is a daily bread for organisms across many different branches of life. Very complex response mechanisms have evolved to tackle such insults. Yeast Saccharomyces cerevisiae is adapted especially well for counteracting oxidative and osmotic stress. These unfavorable conditions usually lead to inhibition of protein synthesis. The GCN2 kinase is thought to be responsible for this phenomenon. General inhibition of protein synthesis is accompanied by an increase in expression of proteins engaging in stress response. Production of these proteins is often preceded by specialized regulatory processes, that operate on various stages of expression. This thesis will try to present the diversity and complexity of the individual regulatory layers.
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The analysis of membrane potential recovery in yeast under CCCP-induced stress
Babuka, David ; Plášek, Jaromír (advisor) ; Sigler, Karel (referee)
The master's thesis is focused on the study of response of the intracellular pH of the yeast cells on various external environments, primarily in a relation to the protonophore carbonyl cyanide m-chlorophenylhydrazone, CCCP. To measure the intracellular pH of the yeast cells we used a genetically coded fluorescent probe the ratiometric pHluorin. Using the method of synchronously scanned fluorescent spectra we were able to measure the intracellular pH of the cells with high precision. As a part of these experiments we also studied the influence of ionic strength of the cell suspensions buffers on the surface potential as well as the influence of the mineral salt KCl on the depolarization of the yeast membranes and cytosolic acidification induced by the protonophore CCCP. We examined the changes of cytosolic pH as such but we also used the measured pH as an indicator of the processes and the state of environment outside the cell. One of the most notable outcomes of this thesis is a new method of monitoring the value of the surface potential of the yeast cells by measuring the titration curves of cytosolic acidification induced by the protonophore CCCP.
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Deconvolution fluorescence microscopy of yeast cells
Štec, Tomáš ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Deconvolution fluorescence microscopy of yeast cells Author: Tomáš Štec Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jarmoír Plášek, CSc., Institute of Physics of Charles Uni- versity Abstract: Fluorescence microscopy presents an fast and cheap alternative to more advanced imaging methods like confocal and electron microscopy, even though it is subject to heavy image distortion. It is possible to recover most of the original distortion-free image using deconvolution in computer image processing. This al- lows reconstruction of 3D structure of studied objects. Deconvolution procedure of NIS Elements AR program undergoes an thorough inspection in this diploma the- sis. It is then applied on restoration of 3D structure of calcofluor stained cell wall of budding yeast Saccharomyces cerevisiae. Changes of the structure of the cell wall during cell ageing are being examined. Cell wall of aged cells shows increased surface roughness and even ruptures at the end of cell life. Keywords: fluorescence, microscopy, deconvolution, NIS Elements AR, calcofluor, yeast, cell wall, ageing
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Extracellular matrix in yeast populations
Novotná, Pavla ; Kuthan, Martin (advisor) ; Dvořáček, Lukáš (referee)
The microorganisms in a natural environment are frequently found in multicellular forms, most commonly in biofilms. Biofilm is characterized as a community of cells living at the interface of two environments, embedded in the extracellular matrix. ECM is a significant component of biofilms in yeast populations. Extracellular matrix acts as a protective barrier and allows cells to survive under adverse conditions and better compete with other microorganisms. It also forms an effective barrier against antibiotics and other harmful substances, what makes biofilms a serious problem in medicine and industry. Formation of the matrix may be influenced by the morphological forms of colonies. Increased formation of ECM is commonly connected in phenotypic switching in response to changes of their living conditions. The compositeion of the ECM is a genus-and species-specific. The main component of the matrix consists of polysaccharides and proteins.
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GAL4/UAS binary system as a toll for the study of gene function
Soukup, Tomáš ; Krylov, Vladimír (advisor) ; Půta, František (referee)
GAL4/UAS system is a bipartite gene engineering tool, enabling ectopic expression in temporal and tissue-specific manner in vivo. Design of this technique is based on a mechanism of gene transcription, which was elucidated of large portion by an experimental study of Saccharomyces cerevisiae regulation of metabolic control circuit for processing galactose. It is possible to generate hundreds of stable transgenic lines by independent incorporation of the gene for the transcription factor Gal4p and its binding sequence (UAS), respectively, by using transposible or specific-sequence integration techniques. An individual organism, manifesting ectopic expression in suitable, adjustable conditions, can be obtained by cross breeding individual of GAL4 lines with individual from UAS line. This thesis represents a synthesis of the basic principles of GAL4/UAS system and its variants, particularly adapted to the needs of genetic manipulation of model organisms Drosophila melanogaster and Danio rerio. Additionally, this text summarizes the connection GAL4/UAS system with other techniques and briefly highlights the potential for practical applications mainly in research area of neurology. Powered by TCPDF (www.tcpdf.org)
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Changes of intracellular pH in yeast cells under stress conditions
Divín, Radek ; Plášek, Jaromír (advisor) ; Krůšek, Jan (referee)
Title: Changes of intracellular pH in yeast cells under stress conditions Author: Radek Divín Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jaromír Plášek, CSc. Abstract: Specific values of intracellular pH (pHi) can affect all biochemical processes in a cell and this phenomenon is closely connected with the degree of importance of changes in the intracellular pH under the stress conditions. In the Master Thesis, the yeast cells Saccharomyces cerevisiae were used as a model of organism eukaryotic cells. Monitoring of intracellular pH of the cells was performed by the method of synchronous fluorescence scan technique of genetically encoded fluorescent probes pHluorin which was located in the cytosol of the cells. The cells were exposed to stress conditions due to the chemical changes in the environment. Consequently, their ability to maintain a stable value of the intracellular pH in various acidic environments was studied in more detail. The attention was also focused on the impact on optimizing of glucose cytosolic pH. The work was centered on the changes in intracellular pH under the influence of the presence of KCl in suspension. Furthermore, the decrease of cytosolic pH of monitored cells by protonophore CCCP was investigated. The effect of stress environment on the...
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