Ni-P based coatings preparation on AZ61 wrought magnesium alloy after heat treatment

Náplní této bakalářské práce je příprava tepelně zpracované tvářené hořčíkové slitiny AZ61 a vyloučení nikl-fosforového povlaku pomocí bezproudé depozice na původní a tepelně zpracovanou hořčíkovou slitinu AZ61. Teoretická část obsahuje dvě hlavní kapitoly. První kapitola se zabývá popisem slitiny AZ61. Druhá část se věnuje bezproudé depozici Ni-P povlaku. Na závěr teoretické části je zpracována rešerše z článků na toto téma. Experimentální část se věnuje charakterizaci a popisu hořčíkové slitiny, jejímu tepelnému zpracování a následnému bezproudému niklování. Byly studovány chemické a mechanické vlastnosti původní AZ61, tepelně zpracované AZ61 a nikl-fosforového povlaku. Pro studium bylo použito metod rastrovací elektronové mikroskopie s energiově-disperzní spektroskopií, optické světelné mikroskopie a měření mikrotvrdosti dle Vickerse.The aim of the bachelor’s thesis is preparation of heat treated wrought magnesium alloy AZ61 and exclusion nickel-phosphorus coating using electroless deposition to the original and heat treated magnesium alloy AZ61. The theoretical part contains two main chapters. The first chapter describes the alloy AZ61. The second part deals with electroless deposition of nickel-phosphorus coating. At the conclusion of the theoretical part is searches on the topic. Experimental part deals with characterization and description of the magnesium alloy, its heat treatment and electroless plating. They were studied chemical and mechanical properties of the original and heat treated AZ61 and nickel-phosphor coating. For the study methods of scanning electron microscopy with energy-dispersive spectroscopy, optical light microscope and Vickers’s microhardness was used.

Comparison of Engineering Education in the United States versus the United Kingdom

There is debate within history books as to exactly who first made the assertion that "America and Britain are two countries divided by a common language." However, no matter who said it first, the statement is an acknowledgement that despite centuries of common goals and aspirations, as well as historic and cultural ties, the two countries have some very distinctive differences. A recent opportunity to teach for two terms at a major university in the United Kingdom, as part of the Fulbright Visiting Scholar Exchange program, provided an opportunity for one U.S. engineering faculty member to examine not only the gaps in our common language, but also the major differences in our university educational systems. Opportunities to associate with four different campuses in Scotland and England provided an excellent chance to compare and contrast how engineering higher education is conducted within these two English-speaking countries on opposite sides of the Atlantic. This paper will examine significant differences which were found in the areas of course conduct, projects, assignments, exams, expectations of student work, coursework assessment and program assessment. Additionally, differences in campus life, pre-collegiate education and general work-life philosophy of both faculty and students will be discussed

Engineering Bacterial Gene Expression: Applications towards Biofuels and Antibiotic Resistance

Nature is rarely static. Organisms live in diverse and stressful environments that necessitate rapid response strategies for survival. Microorganisms have responded to this by evolving bet-hedging, wherein they exhibit constitutively heterogeneous gene expression to maximize fitness across numerous background. The goal of this thesis is to “hijack” this phenomenon using novel gene expression engineering techniques to alter how bacteria respond to their environments, in order to address pressing societal concerns.This begins with a systematic exploration of how bacterial gene expression naturally responds to antibiotics and biofuels. This reveals promising gene candidates for targeted manipulation, for which a library of CRISPR gene expression perturbation devices is constructed. This library is applied to Escherichia coli during exposure to antibiotics and biofuels, and the impact of CRISPR perturbation on growth and fitness is quantified. Many perturbations show significant non-heritable improvements or detriments on growth, indicating the potential of this approach for biofuel and antibiotic applications. To improve the desired bacterial response, individual perturbations are combined in a multiplexed fashion. A significant trend towards lower fitness as more perturbations are combined emerges, which is supported by a systematic exploration of combinatorial perturbation libraries. This trend is correlated to a diminished adaptive potential, suggesting the applicability of multiplexed perturbations for restricting bacterial evolution. In a similar vein, the ability of gene expression perturbations to synergize with antibiotics is explored to identify novel potentiating therapies. Significant gene-drug synergies are characterized and used to potentiate antibiotic treatment in an infection model. Therapeutic peptide nucleic acid molecules are subsequently designed to re-sensitize clinically isolated multidrug-resistant bacteria to treatment. Finally, this thesis expands our available synthetic biology toolkit for manipulating gene expression by outlining novel CRISPR engineering strategies. Deactivated CRISPR proteins are fused with bacterial initiation factor one, and the potential for these constructs to increase translation rates by promoting 30S subunit binding to the ribosome is explored. The design of a smart antibiotic utilizing a CRISPR-holin RNA-based kill switch is also presented. Collectively, this thesis demonstrates the power that manipulating gene expression has in affecting desired phenotypes in bacteria

Bioconversion of cellulose into bisabolene using Ruminococcus flavefaciens and Rhodosporidium toruloides

In this study, organic acids were demonstrated as a promising carbon source for bisabolene production by the non-conventional yeast, Rhodosporidium toruloides, at microscale with a maximum titre of 1055 ± 7 mg/L. A 125-fold scale-up of the optimal process, enhanced bisabolene titres 2.5-fold to 2606 mg/L. Implementation of a pH controlled organic acid feeding strategy at this scale lead to a further threefold improvement in bisabolene titre to 7758 mg/L, the highest reported microbial titre. Finally, a proof-of-concept sequential bioreactor approach was investigated. Firstly, the cellulolytic bacterium Ruminococcus flavefaciens was employed to ferment cellulose, yielding 4.2 g/L of organic acids. R. toruloides was subsequently cultivated in the resulting supernatant, producing 318 ± 22 mg/L of bisabolene. This highlights the feasibility of a sequential bioprocess for the bioconversion of cellulose, into biojet fuel candidates. Future work will focus on enhancing organic acid yields and the use of real lignocellulosic feedstocks to further enhance bisabolene production

Bioconversion of cellulose into bisabolene using Ruminococcus flavefaciens and Rhodosporidium toruloides

In this study, organic acids were demonstrated as a promising carbon source for bisabolene production by the non-conventional yeast, Rhodosporidium toruloides, at microscale with a maximum titre of 1055 ± 7 mg/L. A 125-fold scale-up of the optimal process, enhanced bisabolene titres 2.5-fold to 2606 mg/L. Implementation of a pH controlled organic acid feeding strategy at this scale lead to a further threefold improvement in bisabolene titre to 7758 mg/L, the highest reported microbial titre. Finally, a proof-of-concept sequential bioreactor approach was investigated. Firstly, the cellulolytic bacterium Ruminococcus flavefaciens was employed to ferment cellulose, yielding 4.2 g/L of organic acids. R. toruloides was subsequently cultivated in the resulting supernatant, producing 318 ± 22 mg/L of bisabolene. This highlights the feasibility of a sequential bioprocess for the bioconversion of cellulose, into biojet fuel candidates. Future work will focus on enhancing organic acid yields and the use of real lignocellulosic feedstocks to further enhance bisabolene production

Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate

Microbial production of valuable bioproducts is a promising route towards green and sustainable manufacturing. The oleaginous yeast, Rhodosporidium toruloides, has emerged as an attractive host for the production of biofuels and bioproducts from lignocellulosic hydrolysates. 3-hydroxypropionic acid (3HP) is an attractive platform molecule that can be used to produce a wide range of commodity chemicals. This study focuses on establishing and optimizing the production of 3HP in R. toruloides. As R. toruloides naturally has a high metabolic flux towards malonyl-CoA, we exploited this pathway to produce 3HP. Upon finding the yeast capable of catabolizing 3HP, we then implemented functional genomics and metabolomic analysis to identify the catabolic pathways. Deletion of a putative malonate semialdehyde dehydrogenase gene encoding an oxidative 3HP pathway was found to significantly reduce 3HP degradation. We further explored monocarboxylate transporters to promote 3HP transport and identified a novel 3HP transporter in Aspergillus pseudoterreus by RNA-seq and proteomics. Combining these engineering efforts with media optimization in a fed-batch fermentation resulted in 45.4 g/L 3HP production. This represents one of the highest 3HP titers reported in yeast from lignocellulosic feedstocks. This work establishes R. toruloides as a host for 3HP production from lignocellulosic hydrolysate at high titers, and paves the way for further strain and process optimization towards enabling industrial production of 3HP in the future

Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Lukáš Otoupal Supervisior: RNDr. Hana Vlčková, Ph.D. Title of Thesis: Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum This diploma thesis deals with development of a method for determination of statins in human serum. Extraction by Stage tips pipette tips was chosen and optimized for their extraction. The measurement was performed by the ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometer. The extraction procedure and important mass spectrometer parameters were optimized. Previously developed LC method for determination of statins was used for the analysis. The optimization of the mass spectrometer parameters was started by the selection of precursor ions, consequently the ion source parameters were tuned and finally appropriate ionization mode, SRM transition and collision energy for each analyte were chosen. The suitability of the LC-MS method was verified by the repeatability method (RSD ≤ 1 for the retention time and RSD ≤ 10 for the peak area), linearity (correlation coefficient ≥ 0,997) and sensitivity (LOQ in the range of 5 x 10-10 - 1 x 10- 9 g/ml). Firstly, the optimization employing mixed standard..

Metaverse VR Application

Tato práce shrnuje vývoj Metaverse aplikace od výběru herního jádra, vytváření herních 3D a 2D modelů, herních mechanik, textur a světa tak, aby byly pro hráče zábavné a poutavé. Cílem této práce bylo přiblížit vývoj Metaverse aplikací a vytvoření hratelné aplikace, která se odehrává ve Virtuální realitě.This thesis summarizes the development of Metaverse application from selecting the game core, creating 3D and 2D game models, game mechanics, textures and world to make it fun and engaging for the player. The goal of this thesis was to give an overview of Metaverse app development and the creation of a playable app that takes place in Virtual Reality

Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Lukáš Otoupal Supervisior: RNDr. Hana Vlčková, Ph.D. Title of Thesis: Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum This diploma thesis deals with development of a method for determination of statins in human serum. Extraction by Stage tips pipette tips was chosen and optimized for their extraction. The measurement was performed by the ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometer. The extraction procedure and important mass spectrometer parameters were optimized. Previously developed LC method for determination of statins was used for the analysis. The optimization of the mass spectrometer parameters was started by the selection of precursor ions, consequently the ion source parameters were tuned and finally appropriate ionization mode, SRM transition and collision energy for each analyte were chosen. The suitability of the LC-MS method was verified by the repeatability method (RSD ≤ 1 for the retention time and RSD ≤ 10 for the peak area), linearity (correlation coefficient ≥ 0,997) and sensitivity (LOQ in the range of 5 x 10-10 - 1 x 10- 9 g/ml). Firstly, the optimization employing mixed standard..

Rubidium extraction from cement kiln bypass dust

Náplní diplomové práce je separace rubidia z cementářských bypassových odprašků pomocí kolon na bázi hexakyanoželezantů a pryskyřic.The aim of master's thesis is separation rubidium from cement bypass dust by columns based od hexacyanoferric and resins