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Study of microplastic sorption properties
Hlaváčková, Hana ; Kučerík, Jiří (referee) ; Řezáčová, Veronika (advisor)
There is currently an effort to replace conventional plastics with biodegradable products that will have the ability to replace these substances. One of the possible biodegradable felt, which is currently being studied in detail, is also poly (3-hydroxybutyrate), which is produced by bacteria. This bachelor thesis deals with the sorption of selected substances on poly(3-hydroxybutyrate) microparticles. Substances whose contamination of water sources is a significant problem today have been selected for this work. Specifically, the sorption of the nonsteroidal anti-inflammatory substance ibuprofen, the triazole fungicide tebuconazole and the most common drug caffeine on PHB particles, which fall in the size of the microplastic group, were studied. The method of liquid chromatography with mass detection was used for analyzes.
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Studying the authenticity of coffee of various geographical origins
Flegr, Šimon ; Pořízka, Jaromír (referee) ; Diviš, Pavel (advisor)
This diploma thesis researches coffee authenticity problematice, mainly focusing on the authenticity of geographic origin. In the theoretical part of this work, botanical classification is described as well as production technology and processes. The work also includes chemical composition of coffee, describing the major components and changes during production phases. It describes major production areas of the world, in terms of general description and brief history. Problematics with coffee fraud and its identification are also described. Theoretical part also includes general geological description of 17 studied coffee growing regions. Experimental part is devoted to trace amount analysis of selected elements and volatile compounds. The element analysis was conducted using mass spectrometry or optical emission spectrometry, volatile compounds were determined using gas chromatography combined with mass spectrometry detection. Results were statistically described and analyzed, resulting in several discrimination models based on geographic origin.
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Monitoring of the plasmachemical process using mass spectrometry
Ondra, Zdeněk ; Čáslavský, Josef (referee) ; Čech, Vladimír (advisor)
This bachelor thesis deals with plasma-enhanced chemical vapor deposition (PECVD) and the use of mass spectrometry to monitor the processes in plasma during the deposition of thin film. Tetravinylsilane plasma was used in the process of forming a thin film on the silicon wafer. The background of the spectrometer, the residual gases in the plasma reactor at basic pressure were characterized and the plasma polymerization process was monitored. This process was monitored with increasing effective power (2-150 W). The obtained mass spectra were assigned and described in detail. The plasma species that showed the greatest change were then characterized as a function of time during film deposition.
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Posttranslační modifikace proteinů a jejich role v klíčení rostlin
Berka, Miroslav
Protein posttranslational modification represents a complex extension of proteome diversity and provides a transcription-independent control of cellular processes. This thesis "Protein posttranslational modifications and their role in seed germination" briefly reviews major posttranslational modifications, highlights the role of protein phosphorylation in plant regulatory circuits, and summarises enrichment techniques that are employed in phosphoproteomics. The experimental part describes the development and optimisation of methods that can be utilised in a monitoring of protein posttranslational modifications during barley (Hordeum vulgare L.) seed germination, including a tissue fractionation, proteome equalisation and peptide fractionation techniques. Finally, these methods were employed in an analysis of hydrogen peroxide effect on seed germination. In total, over 1,400 proteins were identified in individual fractions, quantitative profiles were established for more than 881 (~60%) and the validated differentially abundant proteins could indicate a potential mechanism behind the peroxide stimulatory effect. Peptides that would correspond to the identified seed phosphoproteins were mostly inaccessible in this experiment and would require optimised targeted analysis.
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Charakterizace rostlinného proteomu
Petrová, Silvie
Plant leaf tissue proteomics analyses face multiple challenges, including the presence of highly abundant proteins like RuBisCO. This thesis reviews methods in protein analysis and outlines proteome fraction techniques. The experimental part compares the leaf proteome composition of five plants, including three representatives of herbaceous (Arabidopsis thaliana, Solanum lycopersicum, Hordeum vulgare), and two woody plants (Malus domestica and Populus hybrid). Further, peptide fractionations by a high-pH C18 and SCX chromatography, and the role of data processing in proteomics analyses are illustrated.
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Light and its effect on plant proteome
Luklová, Markéta
Light is one of the main environmental factors largely determining plant life. Light provides energy resources for photosynthesis and it is also the most important medium of perception as it represents the main source of information about plant surroundings. My diploma thesis entitled “Light and its effect on plant proteome” briefly summarizes present knowledge about the light perception in plants, its interpretation of light quality, quantity, and direction via photoreceptors. These components trigger a whole range of changes, both on molecular and developmental levels, but the complex molecular mechanisms regulating system of light signalling in plants are far from being resolved. Here, I followed effects of different light parameters on plant proteome, namely, diurnal variations, low- and high-light intensity, spectral composition, and a change in the light period duration. To study these effects two model organisms were selected (i) Arabidopsis thaliana as a representative plant model with the most annotated genome/proteome and (ii) Solanum lycopersicum as a representative plant model significant for agriculture and biotechnology. Proteome changes in response to different light factors were analyzed by LC-MS. In total, the analyses revealed over 100 and 300 light-responsive proteins in Arabidopsis and S. lycopersicum, respectively. Proteomics provided highly sensitive snapshots of protein abundance in response to light and selected experiments have been complemented with a GC-MS metabolite profiling, conductivity measurements, and fluorescence microscopy.
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Fast methods for analysis of migrants from food-contact materials
Vavrouš, Adam ; Čabala, Radomír (advisor) ; Jelínek, Ivan (referee) ; Bajerová, Petra (referee)
(AJ) Food contact materials pose a risk to human health due to a variety of contaminants which they can release into food. These compounds, migrants, include aromatic hydrocarbons, dialkyl phthalates, bisphenols, printing ink photoinitiators, and perfluoroalkyl compounds. The determination of all these substances can be expensive and time-consuming since universally applicable analytical approaches are not available nowadays. This work attempted to develop methods for the simultaneous determination of migrants from all of the mentioned groups in paper-based food contact materials and fatty food. A total of 68 contaminants were studied in paper products using liquid chromatography and gas chromatography coupled with tandem mass spectrometry detection. All analytes were isolated simultaneously using the modified "QuEChERS" method. This method demonstrated acceptable recovery and repeatability for most analytes in the validation study; LOQs ranged from 1.3 to 220 µg/kg. Analysis of 132 real paper products confirmed the occurrence of almost all studied analytes, which were often present in complex mixtures and at concentrations up to 628 mg/kg. A total of 41 potential contaminants were monitored in fatty foods by liquid chromatography with tandem mass detection. Contamination of the mobile phase by...
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Development of components for processing and visualization of proteomic data in KNIME environment
Schneiderová, Anna ; Tauš, Petr (referee) ; Mgr. David Potěšil, Ph.D (advisor)
Proteomics investigates the structure and function of proteins. The proteomic data processing involves several sequential steps, from data transformation and normalization to statistical data evaluation. An integral part of processing proteomic data is also their visualization, which is key for example in the control of input data quality and assessment of statistical evaluation outputs. In the Zbyněk Zdráhal Research Group, a software container is used with KNIME application for reproducible processing of proteomic data. As a part of the bachelor’s thesis, a request survey for new components for KNIME application was carried out. An interactive Volcano plot was chosen for implementation from the proposed components. Python programming language using Dash library was used for implementation. The application can be run independently or as a component of the KNIME environment for working with data from KNIME workflows. The interactive Volcano plot application makes it easy to identify and filter proteomic data.
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Using of techniques of mass spectrometry in the analysis of natural compounds
Vopelková, Alžběta ; Cahlíková, Lucie (advisor) ; Šafratová, Marcela (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Bc. Alžběta Vopelková Supervisor: doc. Ing. Lucie Cahlíková, Ph.D. Title of diploma thesis: Using of techniques of mass spectrometry in the analysis of natural compounds This diploma thesis deals with the topic " Using of techniques of mass spectrometry in the analysis of natural compounds ". The aim of the work was to create a research work and describe the profile of alkaloids in individual genera of the family Amaryllidaceae. The introduction to the theoretical part summarizes the analyzes gas chromatography (GC). Alkaloids have been classified according to their heterocyclic parts and the family Amaryllidaceae has been described. The work was based on professional texts by Czech and foreign authors. Used sources focused on the analysis of alkaloids in bulbs, or in the aboveground parts of plants in this family, using gas chromatography with mass detection (GC-MS). Key words: mass spectometry, analysis, natural compounds
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Clinically relevant protein-protein interactions participating in process of bacterial pathogenesis B. pertusis
Málek, Albert ; Šulc, Miroslav (advisor) ; Černá, Věra (referee)
Whooping cought in human population was strongly suppressed during the 20th century. But in the past few years, the incidence of whooping cough began to rise. The origin of this disease is a pathogenic gram-negative bacteria Bordetella pertussis, which is becoming resistang to currecently used antibiotics or vaccination. B. pertussis attacks human respiratory system. One of it's virulent factors is adenylate cyclase toxin (ACT). It's secreted extracellularly from bacteria and binds to cytoplasmic membrane of host cells and translocates adenylatecyclase domain (dAC) to cytosol. This enzymatic domain is activated by non- covalent interaction with eukaryotic Calmodulin (CaM). After it's activation, dAC in high concentrations synthesizes cAMP, triggering host cell's apoptosis. We studied protein interaction of dAC with CaM by the PIXL method (Photo Induced Cross Linking) and mass spectrometry (MS). Mutant of dAC, with photo-methionine (pM), incorporated in position of Leucine 240 (dACL240pM) was expressed in transformed E.coli cells B834 in mineral medium containing pM. Expressed protein was isolated by affinite chromatography and characterized by MS (determined incorporation of pM was approximately 50 % in the final protein preparation). We performed a photochemical cross-linking with isolated...
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