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Development and application of methods for qualitative and quantitative analysis of amino acid post-translational modifications using liquid chromatography coupled to mass spectrometryLoik, Nikita D. January 2014 (has links)
The 2-oxoglutarate and ferrous ion dependent oxygenases are a super family of enzymes that are involved in a wide range of biological processes regulated trough the mechanism of post-translational modification (PTM). Such biological processes include hypoxia sensing (through regulating HIF transcription), fatty acid metabolism (through carnitine production), transcriptional regulation (through demethylation of histones), and collagen structure formation (through proline and lysine hydroxylation). To understand the underlying mechanisms of such regulatory processes, and to develop clinically useful inhibitors, and thereby regulate these processes in living organisms, requires sensitive methods for monitoring enzyme activity. The use of indirect methods such as quantification of reaction products (14CO2 or succinate) can be problematic, as both products can result from competitive reactions. Alternative direct measurement of substrate modifications using mass spectrometry-based proteomics can be applied; however, (1) for this technique the limit of detection is often prohibitive, (2) the method is best suited for the confirmation of known modifications, rather than for the discovery of new modifications, and (3) sequence coverage may often be only 60%, and therefore many modifications can be missed. The aim of the research presented in this thesis was to develop amino acid analysis and to apply these methods to the identification and quantification of PTMs catalysed by 2-oxoglutarate and ferrous ion dependent oxygenases. A range of LC-MS approaches were investigated including: (1) C18 reversed phase chromatography of quinoline derivatised amino acids, (2) ion paring chromatography, and (3) mixed mode chromatography with either UV, or conventional molecular MS, or isotope ratio mass spectrometry detection. Analysis of the elution patterns for those separation techniques enabled estimation of the retention parameters of modified amino acids and the identification of the modifications, where no standards were available. The most sensitive approach developed employed mixed mode chromatography coupled to isotope ratio mass spectrometry which was optimised for the analysis of modified amino acids. This was shown to have a limit of detection two orders of magnitude lower (0.01μM) than other conventional mass spectrometry techniques. Using amino acid labelling in cell culture, a quantification protocol was developed which employed a non-labelled internal standard and selectively labelled cell culture. The method was shown to be suitable for both very accurate quantification at low concentration levels and metabolic studies, allowing us to track back the modifications to their precursors. The analytical methods developed for amino acid analysis were successfully applied to the analysis of modifications resulting from 2-oxoglutarate and Fe dependent oxygenase activity. Stereochemistry of lysyl hydroxylation in the splicing regulatory protein Luk7L2 by JmjD6 as well as of the self hydroxylation of the JmjD6 was identified. The stereochemistry was shown to be different from that of previously reported for the collagen hydroxyline, hydroxylated by the another member of this enzyme family. mbP4H enzyme was shown to catalyse prolyl hydroxylation of taODD resulting in 4R-hydroxyprolyl. Amino acid analysis was used in order to verify the mechanism of the hBBOX catalysed rearrangement of Mildronate. Using the method developed for the analysis of non-derivatised amino acids the screening of potential substrates of hBBOX enzyme was carried out and two new substrates were identified. The isotope ratio mass spectrometry protocol was applied to the study of histones from cell culture; low levels of hydroxylated and methylated amino acids were quantified. The analytical methods described were developed to complement to the well established proteomics techniques. The methods developed enable investigation into the region- and stereo- chemistry of the modified groups within the modified AA residue and has proved to be a powerful tool of exploratory PTM analysis.
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Transitioning from a Traditional Nursing Home Environment to Green House Homes: What are Stakeholders' Attitudes Toward and Satisfaction With the Small House Care EnvironmentHarrop-Stein, Christine 01 January 2014 (has links)
This dissertation research was designed as two independent research studies. The first study, qualitative, and non-experimental, aimed to examine residents’, family members’, and staff members’ (stakeholders’) satisfaction with, and attitudes toward Green House living one month prior to moving and again at one and three months after moving. Focus groups were the primary method of data collection. Thirty residents and 40 staff members transitioned to one of three Green House homes beginning January, 2013. Data collected began in December, 2012. Following each focus group, tape recordings were transcribed, and coded. Using grounded theory and the constant comparative method of analysis, themes emerged. Pre-move focus group themes revealed that stakeholders were concerned about (a) the quality of care in a system using fewer staff members and (b) the challenges associated with adjusting to a new environment. Post-move focus group themes revealed that (a) stakeholders remained concerned about staffing levels; (b) residents’ had improvements in appetite, socializing, and ambulation; and (c) staff members struggled with autonomous work teams, but preferred the Green House model of care to that of a traditional nursing home. The final model reflects a synthesis of themes from which self-efficacy beliefs were hypothesized. Themes were also linked to existing gerontological theories: Person-Environment Fit, Place-Space, Thriving, and Personhood. The second study, designed to explore the construct validity of the Person-Centered Care Attitude Tool (Per-CCat), consisted of 42 Likert-type questions divided into four sections that align with person-centered care principles. Eighty-six employees of Virginia Mennonite Retirement Community completed the survey; only 70 were analyzed due to missing data. Principal Components Analysis was the analytic approached used for these data. Bartlett’s Test of Sphericity (X2 = 2006.56, p = 0.000) and Keiser-Myers-Olkins measure of sampling adequacy (0.746) indicated that the data were factorable. The final four-factor 34-item solution aligned with the following person-centered care principals: resident autonomy, social interaction and community, work culture, and feelings toward work. Further validations studies of the Per-CCat are necessary. Given the trend in long-term care toward person-centered care, a validated survey will be useful for hiring and educating caregivers and other nursing home personnel.
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The development of a human factors tool for the successful implementation of industrial human-robot collaborationCharalambous, George January 2014 (has links)
Manufacturing organisations have placed significant attention to the potential of industrial human-robot collaboration (HRC) as a means for enhancing productivity and product quality. This concept has predominantly been seen from an engineering and safety aspect, while the human related issues tend to be disregarded. As the key human factors relevant to industrial HRC have not yet been fully investigated, the research presented in this thesis sought to develop a human factors tool to enable the successful implementation of industrial HRC. First, a theoretical framework was developed which collected the key organisational and individual level human factors by reviewing comparable contexts to HRC. The human factors at each level were investigated separately. To identify whether the organisational human factors outlined in the theoretical framework were enablers or barriers, an industrial exploratory case study was conducted where traditional manual work was being automated. The implications provided an initial roadmap of the key organisational human factors that need to be considered as well as the critical inter-relations between them. From the list of individual level human factors identified in the theoretical framework, the focus was given on exploring the development of trust between human workers and industrial robots. A psychometric scale that measures trust specifically in industrial HRC was developed. The scale offers the opportunity to system designers to identify the key system aspects that can be manipulated to optimise trust in industrial HRC. Finally, the results were gathered together to address the overall aim of the research. A human factors guidance tool was developed which provides practitioners propositions to enable successful implementation of industrial HRC.
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Marketingové řízení sportovní značky Nike v ČR / Marketing management of sports brand Nike in the Czech RepublicOndrášková, Adéla January 2015 (has links)
Title: Marketing management of sports brand Nike in the Czech Republic Objectives: The aim of thesis is to describe and evaluate the current marketing management of sports brand Nike in the Czech Republic based on made qualitative analysis. The next point is to detect strengths and weaknesses. Then in the consequence of obtainable knowledge suggest new procedures and implements in marketing which can be used to improve marketing management of the brand. Methods: In this theses there was used a qualitative analysis (descriptive case study), witch was based on document analysis and interview with an employee of marketing department of company. Results: The results showed that marketing management of sports brand Nike in the Czech Republic is on a high level. This fact is mainly a consequence of its global and stable position on a worldwide market. For the reason, that its daughter company Nike CZECH, s. r. o. has been presented on Czech market since 2011, there are many possibilities to improve its marketing management with respect to local specifics. It contains mainly expansion of official shops in the Czech Republic, extensions of brand series, improvements of a communication strategy, sponsorship, changes in an organisation structure and reviews of a brand success. Keywords: brand, marketing,...
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"Vi vinner rättvist" : En studie i lokala sportjournalisters hantering av de journalistiska yrkesidealen. / The home team won fair and square : A qualitative analysis of the level of objectivity in Swedish local sports journalism.Abrahamsson, Joel January 2017 (has links)
In Sweden sports journalism has been critisised among many for lacking professionalism and sports journalists have been accused of being both restricted and biased. Especially in local sports journalism the issue is said to be evenmore prominent.Through a qualitative discourse analysis the main purpose of this essay is to study the standard of objectivity in swedish local sports journalism.Thirty two randomly selected articles written in ten local newspapers after ten games in swedish footballs top divison Allsvenskan were read and analyzed during the study. In relation to the profesional ideals of journalism some major themes of shortcomings were discovered and categorized. The most significant was the lack of hearing both sides. The reporting was very one-sided in a way that most of the time favored the local team. From an objective view questionable reporting was also found when it came to the inclusion and exclusion of certain in-game situations but also in what surrounding circumstances the journalists chose to aim their focus on. Uncertaincies were likewise exposed in the amount of personal opinions being shown by the author.
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Srovnání vybraných herních výkonů hráčů elitní výkonnosti / Comparison of chosen indicators at players of elite performanceKulatka, Ondřej January 2011 (has links)
Title: Comparison of selected variables of the game performance in football. Work objectives: The main objectives of this dissertation is primarily a mutual comparison of two consecutive years of UEFA Champions League 2007/ 2008 and 2008/ 2009, where the greatest effort is to identify new and effective trends in training and scoring offensive action from the "open play", due to the ever improving defense systems and outline new methodological and didactic practices in the training process, which could help the current needs of modern football. Methods: Research is carried out by indirect observation using a video records with all the scored goals that were scored in the Champions League in 2008 / 2009. After an accurate record of all the goals, all goals are evaluated and compared with the previous year. All goals are analyzed from two perspectives, in terms of quantity and quality. The former analysis follows the form of attacks, shooting technique, shooting method, the most common areas of shooting or shooting success due to the game time. Based on these quantitative data are then created interesting graphical illustrations. Qualitative analysis focuses on the breakdown of individual model offensive situations in terms gradual developing of attack and fast counter attack. Results: The results...
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Modely herní součinnosti při zakončování útoků ve fotbale / Models of game cooperations at finishing in footballFajfr, Jakub January 2013 (has links)
Title: Models of game cooperation at finishing in football Objectives: The aim of this diploma thesis is to carry out quantitative and qualitative analysis of football matches played at EURO 2012. Methods: Research was made by indirect observation of football matches. Results: The aim of this thesis was accomplished. The teams were compared in several categories. The hypothesis was confirmed in particular categories and disproved in others. Research questions were answered. Acquired knowledge was used for setting up training exercise. Keywords: game performance, players, game situations, qualitative analysis
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Novináři a ochrana zdrojů ve filmové tvorbě / Journalists and source protection in motion picturesKroupa, Tomáš January 2013 (has links)
This diploma thesis named "Journalists and source protection in motion pictures" focuses on investigative journalists pictured in movies that have originated in Anglo-American language and cultural area. Analyzed are such movies that either have been filmed or have had their opening night in last three decades of the twentieth century or the first decade of twenty first century. From 1970s decade there is one movie analyzed while from all the other decades there are always two movies were being taken into analysis. Theoretical part of this thesis consists first of all of all necessary terms followed by several chapters about history of investigative journalism in English speaking countries. Several important personas and events from the area of investigative journalism from the twentieth century are also mentioned. Political movies from 1960s and 1970s are also briefly mentioned at this point. Briefly discussed are also principles of qualitative analysis method and necessary assumptions. Within the practical part of this thesis every movie is described and analyzed. Specific qualitative analysis method is the same in case of all the analyzed movies and results are discussed in specific subchapters by the same structure so all movies can be easily compared and cross-analyzed. Detailed descriptions...
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Magazín Forbes perspektivou politické ekonomie komunikace: Kritická analýza roku existence titulu na českém mediálním trhu / Forbes magazine in terms of the political economy of communication: a critical analysis of a year of existence of the czech media marketBendlová, Klára January 2015 (has links)
Master thesis concerns itself with Czech edition Forbes magazine. Regarding the global media market today, journal Forbes without a doubt belongs to the fully respected periodical. Magazine Forbes has gained its reputation thru regular publishing business charts. Concept of this magazine was established on the stories of successful people. Thesis examines introduction of the journal on the Czech market and its first year of existence in the conditions of domestic media market. This view is based on political economical communication. The emphasis, in the Czech issue Forbes magazine, is put on the rise of success and identities thru political economical communication and its confrontation with cultural studies. In the frame of wider politico-economic context is part of the thesis dedicated to original American edition of the magazine, whose publisher owns the rights for publishing the magazine in Czech Republic. The main research on the political economical bases related to Forbes magazine is characterised by the production of this magazine, its position and goals on the market, strategy to achieve and target audience. Thesis contains quantitative analysis of the magazine contents and qualitative analysis editorials editor in chief. That should have provided deeper view into the meanings and identities in...
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Dynamics of biotechnology entrepreneurship in South Africa and BrazilAlagbaoso, Manessah Obinali 04 August 2014 (has links)
Biotechnology entrepreneurship is a relatively new and distinct field of
entrepreneurship. Most current empirical research is conducted in the developed
economies and cannot be directly extrapolated to the developing economies. The
paucity of empirical research and the lack of a conceptual framework for
biotechnology entrepreneurship constitute gaps that this research addressed through
the development of a proposed theoretical framework of biotechnology
entrepreneurship based on empirical research conducted within the context of the
developing economies of South Africa and Brazil.
The current methodological approaches to research in biotechnology
entrepreneurship predominantly make use of a nomothetic philosophical approach
and employs quantitative methods. Current research is also often based on surveys
conducted within one developing economy context. Consequently, few studies in
biotechnology entrepreneurship use the qualitative multiple case study approach.
This methodological gap is addressed in this research through the use of qualitative
multiple case studies, in the idiographic philosophical tradition, in two developing
economies; South Africa and Brazil. The data collection process included in-depth
interviews, documents review and observations, which improved the quality of the
research through data triangulation.
Ten themes were identified, which formed the basis for developing the proposed
theoretical framework. In addition, seven factors that influence the process of
biotechnology entrepreneurship in South Africa and Brazil were identified as
regulation; funding; infrastructure; skills; entrepreneurial and commercialisation
capabilities; market for biotechnology products; and social development.
This research shows that the individual-opportunity nexus of entrepreneurship does
not entirely hold for biotechnology entrepreneurship in South Africa and Brazil.
Instead, there is a nexus of research and development; and a governmentincentivised
environment that is conducive for biotechnology entrepreneurship.
The policy implications of these dynamics in South Africa and Brazil; as well as implications for the other stakeholders in the biotechnology industry are articulated as being linked to the control of the factors that influence biotechnology entrepreneurship by the various stakeholders. Hence, the implications for government are predominantly linked to regulation and infrastructure; and the implications for the other stakeholders are predominantly linked to funding and skills.
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