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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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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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Developing a methodological geographic information system framework to augment identification of future risk of anomalous dwelling firesDean, Emma January 2015 (has links)
This thesis outlines research completed in partnership between Merseyside Fire and Rescue Service and Liverpool John Moores University. The aim of the research was to investigate ways to develop and implement a bespoke Geographic Information System framework that could be used to identify risk of future anomalous accidental dwelling fires. This thesis outlines the techniques used to develop the framework and its application. In particular, the thesis presents an understanding of accidental dwelling fire causal factors and how data related to these can be incorporated into a model for identifying risk and targeting initiatives relative to the risk. The thesis also investigates two strands of customer insight developed for Merseyside Fire and Rescue Service. These are community profiles, based on a cluster analysis approach, to understand risks present within communities and the vulnerable person index, which identifies individuals most at risk from fire using data shared through information sharing agreements. Nationally recognised risk modelling toolkits, such as the Fire Service Emergency Cover toolkit do not utilise local information or have the ability to identify risk to an individual level. There is a need for this intelligence to be able to proactively target services, such as the Home Fire Safety Check. This paper also discusses some of the key operational and strategic areas that benefit from this information and presents some case studies related to the application of the research.
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Qualitative and quantitative analysis of systems and synthetic biology constructs using P systemsKonur, Savas, Gheorghe, Marian, Dragomir, C., Mierla, L.M., Ipate, F., Krasnogor, N. 04 August 2014 (has links)
Yes / Computational models are perceived as an attractive alternative to mathematical models (e.g., ordinary differential equations). These models incorporate a set of methods for specifying, modeling, testing, and simulating biological systems. In addition, they can be analyzed using algorithmic techniques (e.g., formal verification). This paper shows how formal verification is utilized in systems and synthetic biology through qualitative vs quantitative analysis. Here, we choose two well-known case studies: quorum sensing in P. aeruginosas and pulse generator. The paper reports verification analysis of two systems carried out using some model checking tools, integrated to the Infobiotics Workbench platform, where system models are based on stochastic P systems. / EPSRC
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Mediální reprezentace utváření komunální politiky (na příkladu sporu o grantování pražských divadel v letech 2007 až 2008) / Mediarepresentation of forming local policyŠvubová, Jana January 2013 (has links)
The diploma thesis on the "Media representation of forming of local policy (on the example of the financing of the Prague theatres)" suggests that mass media contribute to the forming of local policy by selective reporting of the politics. This fact is illustrated on the example of forming grant policy of the capital city of Prague. The analysed media are the daily papers MF Dnes, Lidové noviny, Právo and the televisions TV Nova and Česká televize in 2007 and 2008. The theoretical part of the diploma thesis introduces the relation between mass media and reality, the effects of media and the factors influencing forming of the news content. The political communication is discussed; this phenomenon is introduced together with the personalisation of politics and the mediatisation. Furthermore the paper deals with the agenda- setting theory and its relation to framing theory and the theory of news values. Then the development of the dispute is given, the most important moments are emphasized. The research part of the diploma thesis includes results of the quantitative content analysis. The aim of this research was to discover the media image of the dispute and to confirm or to disprove the hypothesis (i.e. the mass media were not impartial). The qualitative frame analysis shows the ways the dispute was...
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Estudo do conversor D : an?lises qualitativa e quantitativa, crit?rios de dimensionamento e modelagem em espa?o de estadosDutra, S?lvio Cesar Antunes 31 August 2018 (has links)
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Previous issue date: 2018-08-31 / Power converters are widely used in the context of power electronics. In the same way that transformers have a fundamental role in applications involving AC circuits, power converters modify voltage magnitudes in DC circuits. These devices can either present an output voltage higher than the input voltage, acting as a Boost converter, or lower, acting, in this case, as a Buck converter. The input and output signals can also present either a voltage source or a current source behavior, depending on the topology. This work presents a systematic study of the D converter operating in the continuous conduction mode, in permanent regime, including the qualitative and quantitative analyzes, resulting in a methodology of dimensioning of the converter, as well as its modeling in space of states, which makes feasible studies of control strategies for it. Although this is not a new topology, this converter has recently been reintroduced due to its ability to increase efficiency in the energy conversion process, especially in photovoltaic systems. However, this research is fully justified given the lack of similar work for the D converter topology. This study was validated from tests performed in the PSIM simulation software, in addition to MATLAB / SIMULINK. A laboratory prototype was specially constructed for validation purposes in a rapid prototyping platform, as well as the consequent implementation in a PCI, for use in future works. / Os conversores est?ticos s?o amplamente utilizados no contexto da eletr?nica de pot?ncia. Da mesma forma que os transformadores t?m papel fundamental nas aplica??es que envolvem circuitos alimentados em corrente alternada, os conversores est?ticos realizam a transforma??o de magnitudes de tens?o em circuitos de corrente cont?nua. Tais dispositivos podem tanto apresentar em sua sa?da um valor de tens?o superior em rela??o ao sinal de entrada, atuando como elevadores, quanto inferior, atuando, neste caso, como redutores. Estes tamb?m podem apresentar suas sa?das ou entradas com um comportamento de fonte de tens?o ou corrente, dependendo da topologia utilizada. Este trabalho apresenta um estudo sistematizado do conversor D operando no modo de condu??o cont?nua, em regime permanente, incluindo as an?lises qualitativa e quantitativa, resultando em uma metodologia de dimensionamento do conversor, bem como a sua modelagem em espa?o de estados, o que viabiliza estudos de estrat?gias de controle para o mesmo. Apesar de n?o se tratar de uma nova topologia, recentemente este conversor foi reintroduzido, devido a sua capacidade de aumentar a efici?ncia no processo de convers?o de energia, especialmente em sistemas fotovoltaicos. Esta pesquisa encontra justificativa dada a inexist?ncia de trabalhos semelhantes especificamente para a topologia do conversor D. Este estudo foi validado a partir de ensaios realizados no software de simula??o PSIM, al?m do MATLAB/SIMULINK. Um prot?tipo de laborat?rio foi especialmente constru?do para fins de valida??o em uma plataforma de prototipagem r?pida, bem como a consequente implementa??o do mesmo em uma PCI, para utiliza??o em trabalhos futuros.
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Cross-cultural study on decision making of German and Indian university studentsTipandjan, Arun 04 June 2010 (has links) (PDF)
The dissertation consists of an introduction and three empirical articles. The introduction gives the theoretical background, integrates the three articles, and elaborates on future research questions. The first article investigates the important decision in the lives of German and Indian university students to identify the important areas of decision making. The second article examines the structure of real life decision making and reveals the underlying factors of five major decision areas. The third article investigates the similarities and differences on decision making between German and Indian students using prior qualitative findings in a large quantitative survey.
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Modélisation, analyse et conception de protocoles MAC multi-canaux dans les réseaux sans fil / Modeling, analysis and design of multi-channel MAC protocols in wireless networksEl Fatni, Abdelaziz 03 June 2013 (has links)
L’utilisation des communications multi-canaux au niveau des protocoles MAC (MAC multi-canaux) dans les réseaux sans fil peut améliorer significativement le débit du réseau par rapport aux protocoles MAC mono-canal en permettant à plusieurs transmissions concurrentes d’avoir lieu simultanément sans interférence sur différents canaux non recouverts. Cependant, l’environnement multicanaux ajoute une nouvelle dimension aux protocoles MAC qui nécessite un nouveau mécanisme appelé "le mécanisme d’allocation des canaux" pour coordonner l’utilisation des canaux. Le rôle de ce mécanisme est de permettre aux nœuds de s’accorder sur le canal qui doit être utilisé et à quel moment pour la transmission de données dans un contexte distribué. L’absence de ce mécanisme ou une mauvaise coordination de l’utilisation des canaux peut produire l’effet inverse de celui recherché. Ainsi, l’efficacité d’un protocole MAC multi-canaux dépend strictement de l’efficacité du mécanisme d’allocation des canaux, qui par ailleurs, doit offrir les garanties nécessaires au fonctionnement de la communication multi-canaux. Dans cette thèse, nous nous intéressons à la modélisation, l’analyse et la conception de nouveaux protocoles MAC multi-canaux dans les réseaux sans fil. Dans un premier temps, nous nous focalisons sur la spécification formelle et la vérification des propriétés fondamentales inhérentes aux activités de l’allocation des canaux et son impact sur le transfert de données des principales approches MAC multi-canaux proposées dans la littérature. Dans un second temps, nous visons à proposer, modéliser et à analyser formellement deux nouveaux protocoles MAC multicanaux résultants de l’analyse qualitative réalisée. Enfin, une étude quantitative et comparative est menée afin d’analyser les performances des protocoles proposés en terme de débit et d’impact du temps de commutation entre les canaux. / The use of multi-channel communications in MAC protocols (multi-channel MAC) for wireless networks can significantly improve the network throughput compared with single channel MAC protocols since concurrent transmissions can take place simultaneously on distinct non-overlapping channels without interfering. However, due to the multi-channel environment, a novel issue must be addressed in MAC protocols, which is called "the channel assignment mechanism" in order to coordinate the channels access. The role of this mechanism is to enable nodes to agree on which channel should be used and when for data transmission in a distributed way. The absence of this mechanism or a bad coordination of the channel usage can incur the opposite effect to that intended. Thus, the efficiency of a multi-channel MAC protocol depends strictly on the efficiency of the channel assignment mechanism, which also must provide the guarantees necessary to the good behavior of the multi-channel communications. In this thesis, we focus on the modeling, analysis, and design of new multi-channel MAC protocols in wireless networks. Firstly, we focus on the formal specification and verification of fundamental properties inherent to the activities of the channel assignment and its impact on the data transfer for the main multi-channel MAC approaches proposed in the literature. Secondly, we aim to propose, model and analyze formally two new multichannel MAC protocols, resulting from the qualitative analysis conducted. Finally, a quantitative and comparative study is conducted to analyze the performance of the proposed protocols in terms of the throughput and the impact of the switching time between channels.
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The relevance of preoperative ultrasound cervical mapping in thyroid cancerKocharyan, Davit 12 1900 (has links)
Pendant les trente dernières années, le taux d'incidence du cancer de la thyroïde chez l'homme et la femme a considérablement augmenté partout dans le monde. Cependant, on estime que d'ici à 2019 le cancer de la thyroïde deviendra le troisième cancer le plus répandu chez les femmes dans tous les groupes d'âge en raison de la tendance d’augmentation plus dramatique chez elles. En général, il n'y a aucune raison claire qui explique l'augmentation mondiale de l'incidence du cancer de la thyroïde et il est émis l'hypothèse que cette recrudescence de l'incidence a une étiologie multifactorielle. Bien qu'il soit clair que le progrès technique des modalités de l’imagerie diagnostique telle que l'échographie peut amener à une augmentation du taux de détection du cancer de la thyroïde secondaire au sur-diagnostic des maladies sous-cliniques, il existe des preuves fortes indiquant une vraie augmentation du cancer de la thyroïde.
La cartographie cervicale échographique préopératoire est un outil important dans l'algorithme diagnostic du cancer de la thyroïde. Elle aide à identifier l’étendue des métastases ganglionnaires cervicales afin de guider la dissection chirurgicale anticipée. La dissection chirurgicale du cou orientée selon les compartiments anatomiques et guidée par la cartographie cervicale échographique peut amener à une réduction des risques des complications postopératoires et des récidives tumorales locorégionales.
Nous avons effectué une analyse qualitative et quantitative de la cartographie cervicale échographique afin d'évaluer la fiabilité diagnostique de ce test. Nos résultats ont démontré une valeur prédictive positive assez élevée de cette modalité diagnostique ainsi q’une association quantitative forte entre les données de la cartographie échographique et les résultats de l’histopathologie. Nous suggérons que l’utilisation de la cartographie cervicale échographique cible les patients présentant un risque plus important d’une maladie persistante / récidivante. / Over the last 30 years, the incidence rate of thyroid cancer has drastically increased in both genders all over the world. However, due to a more dramatic pattern in females, it is estimated that by 2019 it will become the third most prevalent cancer in women of all ages.
Overall, there are no clear reasons behind the worldwide increase in thyroid cancer incidence and it is hypothesized that this upsurge has a multifactorial etiology. Despite the fact that recent advances in imaging modalities such as ultrasound can lead to thyroid cancer overdiagnosis by improving the detection rate for subclinical disease, there is strong evidence indicating a true increase in the occurrence of thyroid cancer as well.
Preoperative ultrasound cervical mapping, an important tool in the diagnostic algorithm of thyroid cancer, helps to identify metastatic spread in cervical lymph nodes and guides the surgeon for subsequent surgical dissection. Compartment oriented neck dissection directed by ultrasound mapping decreases locoregional tumor recurrence and lowers the risk of postsurgical complications.
We conducted a qualitative and quantitative analysis of ultrasound mapping to evaluate this test’s diagnostic reliability. Our results demonstrated that the positive predictive value of this diagnostic modality was sufficiently high and that there was a strong quantitative association between ultrasound mapping and histopathology results. We therefore recommend that ultrasound mapping be used to target patients with a higher risk of persistent or recurrent thyroid cancer.
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Desenvolvimento de metodologia analítica específica e seletiva para a determinação do nitrato de econazol em cremes / Development of a specific and selective analytical methodology for the determination of econazole nitrate in creamsGaldos, Angel Arturo Gaona 18 April 2008 (has links)
Neste trabalho realizaram-se provas de identificação qualitativa e quantitativa do nitrato de econazol (matéria-prima); ensaios de caracterização térmica do nitrato de econazol, excipientes e formulação farmacêutica (creme), assim como desenvolvimento de metodologia analítica específica e seletiva para a quantificação do nitrato de econazol na formulação farmacêutica de creme. A caracterização térmica dos excipientes foi realizada usando-se a Termogravimetria (TG). Para a caracterização térmica do nitrato de econazol e da formulação foram empregadas a Termogravimetria, a Termogravimetria Derivada (TG/DTG) e a Calorimetria Exploratória Diferencial (DSC). Na caracterização térmica da formulação foram identificados e quantificados dois tipos de água com interações diferentes com a matriz. No desenvolvimento da metodologia específica utilizou-se a eletroforese capilar em zona, método linear com concentrações entre 80 e 120 µg mL-1, com coeficiente de correlação de 0,9995; precisão calculada como desvio padrão relativo (DPR) menor de 2%; exatidão do método comprovada mediante teste de recuperação, obtendo-se valores de 100±2,5%; limites de detecção e quantificação 1,853 µg.mL-1 e 5,617 µg.mL-1, respectivamente, comprovando-se também que o método é robusto. No desenvolvimento da metodologia seletiva foram usadas a cromatografia líquida de alta eficiência com eluição em gradiente (CLAE) e a eletroforese capilar pelo método da cromatografia eletrocinética micelar (MEKC). Nestes métodos foram separados o nitrato de econazol, as impurezas relatadas na literatura do nitrato de econazol (4-álcool clorobenzílico, alfa-2,4-diclorofenil-1 H¬imidazol-1-etanol) e os conservantes presentes na formulação (metilparabeno e propilparabeno). O método seletivo desenvolvido por CLAE foi linear para todas as moléculas, com coeficientes de correlação maiores de 0,99. A precisão calculada para o nitrato de econazol como DPR foi menor de 2%; a exatidão para o nitrato de econazol foi comprovada mediante teste de recuperação, obtendo-se valores de 100±2%; comprovou-se também que o método é robusto e que poderia ser aplicado para a quantificação de cada um destes cinco compostos. No método seletivo desenvolvido por MEKC se obteve tempo menor em comparação ao método por CLAE. / In this work, were performed qualitative and quantitative identitication econazole nitrate (raw material); thermal characterization of econazole nitrate, excipients and formulation (cream). There also were developed and validated specific and selective analytical methodology for quantitative determination of econazole nitrate in creams. The thermal characterization of drug product excipients was performed using the Thermogravimetry (TG). For the thermal characterization of econazole nitrate drug and their formulation were used the Thermogravimetry, Derivative Thermogravimetry (TG/DTG) and Differential Scanning Calorimetry (DSC). From cream formulation were identified and quantified two kinds of water which have different interactions with matrix. As regards lo developing of specific methodology was used the capillary zone electrophoresis (CZE). The method was linear, with a correlation coefficient of 0.9995 in the concentration range of 80 to 120 µg.mL-1 of econazole nitrate, precision calculated as relative standard deviation (RSD) was less than 2%, accuracy calculated percentage of recovery for commercial sample was of 100 ± 2.5%, detection and quantitation limits were 1.853 µg.mL-1 and 5.617 µg.mL-1, respectively. The CZE method showed to be robust. For developing of selective methodology were used high performance liquid chromatography (HPLC) with gradient elution, and micellar electrokinetic chromatography (MEKC) methods. Econazole nitrate, related impurities the drug (4-Chlorobenzyl alcohol, alpha-2,4-dichlorophenyl-1H - imidazole-1-ethanol) and preservatives (methylparaben, propylparaben) present in the drug product were separated. The HPLC method was linear for all analytes, the correlation coefficients were higher than 0.99. The precision for nitrate econazol in terms of RSD was less than 2%, the percentage of recovery for commercial sample of econazole nitrate was 100±2%, the method is robust and could be applied for the quantitation of the five substances cited above. With the MEKC method developed the five substances were separated less time comparated with HPLC method.
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