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BARBOSA, Ana Clara de Oliveira Ferraz. "Avaliação de critérios de compatibilidade entre pares de primers para otimização de sistemas multiplex de genotipagem". Universidade Federal de Goiás, 2010. http://repositorio.bc.ufg.br/tede/handle/tde/1291.
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Previous issue date: 2010-01-12The progress of Molecular Biology and Genetics provided the appearance of several molecular markers that detect the genetic polymorphism directly at DNA. Among these markers are the microsatellites (SSR), which are distinguished by their high degree of polymorphism. The use of these markers for individual genotyping has evolved into multiplex systems, which allow many SSR fragments to be detected and analyzed simultaneously. Currently there are several articles in literature discussing the criteria to be used in the primer design for use in PCR, as well as various softwares are available for this end. However, there are few studies and tools for the analysis of compatibility between pairs of primers for use in multiplex systems, where multiple fragments are simultaneously amplified using PCR. This paper evaluated different criteria for compatibility between pairs of primers. A set of 74 combinations of pairs of primers, involving the amplification of 94 SSR loci were evaluated in duplex systems. The same combinations were evaluated according to different criteria, including the degree of complementarity between primers, the magnitude of differences of denaturation temperatures (Tm) and the tendency to annealing between pairs of primers based on the Gibbs free energy resulting from the association between them. The comparison between the different criteria allowed the identification of a set of criteria with positive predictive value equal to 94%. These criteria were implemented for use in a software called Multiplexer, which from the analysis in sequence of pairs of primers, suggests compatible combinations for use in multiplex genotyping systems. Using this tool can significantly reduce the costs related to laboratory activities for genotyping using PCR.
Os avanços da Biologia Molecular e da Genética proporcionaram o surgimento de diversos marcadores moleculares que detectam o polimorfismo genético diretamente no DNA. Entre estes marcadores se encontram os microssatélites (SSR), que se destacam pelo seu elevado grau de polimorfismo. O uso desses marcadores para fins de genotipagem individual tem evoluído para sistemas multiplex, os quais permitem que vários fragmentos SSR sejam detectados e analisados simultaneamente. Atualmente são abundantes na literatura artigos que discutem os critérios a serem utilizados no desenho de pares de primers para aplicação em PCR, bem como estão disponíveis diversos softwares para este fim. No entanto, ainda são escassos os estudos e ferramentas destinados à análise de compatibilidade entre pares de primers para aplicação em sistemas multiplex, onde vários fragmentos são amplificados simultaneamente por PCR. Neste trabalho são avaliados diferentes critérios de compatibilidade entre pares de primers. Um conjunto de 74 combinações de pares de primers, envolvendo a amplificação de 94 locos SSR foram avaliados em sistemas duplex. As mesmas combinações foram avaliadas segundo diferentes critérios, incluindo o grau de complementariedade entre primers, magnitude das diferenças de temperaturas de desnaturação (Tm) e a tendência ao anelamento entre pares de primers com base na energia livre de Gibbs resultante da associação entre eles. A comparação entre os diferentes critérios permitiu a identificação de um conjunto de critérios com valor preditivo positivo igual a 94%. Estes critérios foram implementados para utilização em um software denominado Multiplexer, que a partir da análise de sequências de pares de primers, sugere combinações compatíveis para a utilização em sistemas de genotipagem multiplex. O uso dessa ferramenta pode reduzir consideravelmente os custos laboratoriais relativos às atividades de genotipagem utilizando PCR.
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Dahl, Fredrik. "Selector Technology : For Multiplex DNA Analysis". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5921.
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Ren, Haolin. "Visualizing media with interactive multiplex networks". Thesis, Bordeaux, 2019. http://www.theses.fr/2019BORD0036/document.
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Les flux d’information suivent aujourd’hui des chemins complexes: la propagation des informations, impliquant éditeurs on-line, chaînes d’information en continu et réseaux sociaux, emprunte alors des chemins croisés, susceptibles d’agir sur le contenu et sa perception. Ce projet de thèse étudie l’adaptation des mesures de graphes classiques aux graphes multiplexes en relation avec le domaine étudié, propose de construire des visualisations à partir de plusieurs représentations graphiques des réseaux, et de les combiner (visualisations multi-vues synchronisées, représentations hybrides, etc.). L’accent est mis sur les modes d’interaction permettant de prendre en compte l’aspect multiplexe (multicouche) des réseaux. Ces représentations et manipulations interactives s’appuient aussi sur le calcul d’indicateurs propres aux réseaux multiplexes. Ce travail est basé sur deux jeux de données principaux: l’un est une archive de 12 ans de l’émission japonaise publique quotidienne NHK News 7, de 2001 à 2013. L’autre recense les participants aux émissions de télévision/radio françaises entre 2010 et 2015. Deux systèmes de visualisation s’appuyant sur une interface Web ont été développés pour analyser des réseaux multiplexes, que nous appelons «Visual Cloud» et «Laputa». Dans le Visual Cloud, nous définissons formellement une notion de similitude entre les concepts et les groupes de concepts que nous nommons possibilité de co-occurrence (CP). Conformément à cette définition, nous proposons un algorithme de classification hiérarchique. Nous regroupons les couches dans le réseau multiplexe de documents, et intégrons cette hiérarchie dans un nuage de mots interactif. Nous améliorons les algorithmes traditionnels de disposition de mise en forme de nuages de mots de sorte à préserver les contraintes sur la hiérarchie de concepts. Le système Laputa est destiné à l’analyse complexe de réseaux temporels denses et multidimensionnels. Pour ce faire, il associe un graphe à une segmentation. La segmentation par communauté, par attribut, ou encore par tranche temporelle, forme des vues de ce graphe. Afin d’associer ces vues avec le tout global, nous utilisons des diagrammes de Sankey pour révéler l’évolution des communautés (diagrammes que nous avons augmentés avec un zoom sémantique). Cette thèse nous permet ainsi de parcourir trois aspects (3V) des plus intéressants de la donnée et du BigData appliqués aux archives multimédia: Le Volume de nos données dans l’immensité des archives, nous atteignons des ordres de grandeurs qui ne sont pas praticables pour la visualisation et l’exploitation des liens. La Vélocité à cause de la nature temporelle de nos données (par définition). La Variété qui est un corollaire de la richesse des données multimédia et de tout ce que l’on peut souhaiter vouloir y investiguer. Ce que l’on peut retenir de cette thèse c’est que la traduction de ces trois défis a pris dans tous les cas une réponse sous la forme d’une analyse de réseaux multiplexes. Nous retrouvons toujours ces structures au coeur de notre travail, que ce soit de manière plus discrète dans les critères pour filtrer les arêtes par l’algorithme Simmelian backbone, que ce soit par la superposition de tranches temporelles, ou bien que ce soit beaucoup plus directement dans la combinaison d’indices sémantiques visuels et textuels pour laquelle nous extrayons les hiérarchies permettant notre visualisationNowadays, information follows complex paths: information propagation involving on-line editors, 24-hour news providers and social medias following entangled paths acting on information content and perception. This thesis studies the adaptation of classical graph measurements to multiplex graphs, to build visualizations from several graphical representations of the networks, and to combine them (synchronized multi-view visualizations, hybrid representations, etc.). Emphasis is placed on the modes of interaction allowing to take in hand the multiplex nature (multilayer) of the networks. These representations and interactive manipulations are also based on the calculation of indicators specific to multiplex networks. The work is based on two main datasets: one is a 12-year archive of the Japanese public daily broadcast NHK News 7, from 2001 to 2013. Another lists the participants in the French TV/radio shows between 2010 and 2015. Two visualization systems based on a Web interface have been developed for multiplex network analysis, which we call "Visual Cloud" and "Laputa". In the Visual Cloud, we formally define a notion of similarity between concepts and groups of concepts that we call co-occurrence possibility (CP). According to this definition, we propose a hierarchical classification algorithm. We aggregate the layers in a multiplex network of documents, and integrate that hierarchy into an interactive word cloud. Here we improve the traditional word cloud layout algorithms so as to preserve the constraints on the concept hierarchy. The Laputa system is intended for the complex analysis of dense and multidimensional temporal networks. To do this, it associates a graph with a segmentation. The segmentation by communities, by attributes, or by time slices, forms views of this graph. In order to associate these views with the global whole, we use Sankey diagrams to reveal the evolution of the communities (diagrams that we have increased with a semantic zoom). This thesis allows us to browse three aspects of the most interesting aspects of the data miming and BigData applied to multimedia archives: The Volume since our archives are immense and reach orders of magnitude that are usually not practicable for the visualization; Velocity, because of the temporal nature of our data (by definition). The Variety that is a corollary of the richness of multimedia data and of all that one may wish to want to investigate. What we can retain from this thesis is that we met each of these three challenges by taking an answer in the form of a multiplex network analysis. These structures are always at the heart of our work, whether in the criteria for filtering edges using the Simmelian backbone algorithm, or in the superposition of time slices in the complex networks, or much more directly in the combinations of visual and textual semantic indices for which we extract hierarchies allowing our visualization
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Wegner, Margret [Verfasser]. "Arthrogryposis multiplex congenita : Eine Klassifikation / Margret Wegner". Aachen : Shaker, 2007. http://d-nb.info/117052771X/34.
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Rios, Villanueva Xavier. "Toward Multiplex Genome Engineering in Mammalian Cells". Thesis, Harvard University, 2013. http://dissertations.umi.com/gsas.harvard:11179.
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Given the explosion in human genetic data, new high-throughput genetic methods are necessary for studying variants and elucidating their role in human disease. In Chapter I, I will expand on this concept and describe current methods for genetically modifying human cells. In E. coli, Multiplex Automatable Genome Engineering (MAGE) is a powerful tool that enables the targeting of multiple genomic loci simultaneously with synthetic oligos that are recombined at high frequencies in an optimized strain. MAGE as a method has two components: organism-specific optimization of oligo recombination parameters and a protein capable of increasing recombination frequencies.
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Battiston, Federico. "The structure and dynamics of multiplex networks". Thesis, Queen Mary, University of London, 2017. http://qmro.qmul.ac.uk/xmlui/handle/123456789/30631.
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Network science has provided useful answers to research questions in many fields, from biology to social science, from ecology to urban science. The first analyses of networked systems focused on binary networks, where only the topology of the connections were considered. Soon network scientists started considering weighted networks, to represent interactions with different strength, cost, or distance in space and time. Also, connections are not fixed but change over time. This is why in more recent years, a lot of attention has been devoted to temporal or time-varying networks. We now entered the era of multi-layer networks, or multiplex networks, relational systems whose units are connected by different relationships, with links of distinct types embedded in different layers. Multiplexity has been observed in many contexts, from social network analysis to economics, medicine and ecology. The new challenge consists in applying the new tools of multiplex theory to unveil the richness associated to this novel level of complexity. How do agents organise their interactions across layers? How does this affect the dynamics of the system? In the first part of the thesis, we provide a mathematical framework to deal with multiplex networks. We suggest metrics to unveil multiplexity from basic node, layer and edge properties to more complicated structure at the micro- and meso-scale, such as motifs, communities and cores. Measures are validated through the analysis of real-world systems such as social and collaboration networks, transportation systems and the human brain. In the second part of the thesis we focus on dynamical processes taking place on top of multiplex networks, namely biased random walks, opinion dynamics, cultural dynamics and evolutionary game theory. All these examples show how multiplexity is crucial to determine the emergence of unexpected and instrinsically multiplex collective behavior, opening novel perspectives for the field of non-linear dynamics on networks.
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Allen, James. "The public goods game on multiplex networks". Thesis, University of Surrey, 2018. http://epubs.surrey.ac.uk/845834/.
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Cooperation is acting in the interests of one’s social group, often at a cost to yourself. When the level of cooperation is observed in the laboratory, people cooperate more often, and at higher levels than are predicted by standard theories. In this thesis I find two novel ways in which cooperation on multilayered populations is increased. These models contribute to an understanding of how people cooperate in real-world social situations, and help us to explain why people cooperate as much as they are observed to do. In each study I model the tension between the individual and the group using the public goods game. This game is played on a structured population defined by a multilayered network. Each layer represents a different sphere of influence on the player’s decision to cooperate or defect. The first model studies the effect of a player choosing whether to cooperate or defect on either all layers simultaneously (synchronously) or on one layer at a time (asynchronously). Updating asynchronously leads to increased cooperation across a number of different parameter regimes. This demonstrates a new way in which cooperation can be increased in a system with multiple influences, and also helps to understand exactly why cooperation is increased in multilayered systems. Inspired by empirical examples, the second model adds to the standard model of the public goods game on networks in two ways. The first is to include conditional cooperators, and the second is the addition of a layer of social influence. This combination of economic and social influence has not been considered in previous models of the public goods game, and I find that this additional layer of influence results in high levels of cooperation. In the final chapter, I study these dynamics on more realistic network structures, with results echoing empirical findings under certain parameters.
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Vorce, Richard G. "A TUNABLE MULTIPLEX DISCRIMINATOR WITH DIGITAL OUTPUT". International Foundation for Telemetering, 1985. http://hdl.handle.net/10150/615551.
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International Telemetering Conference Proceedings / October 28-31, 1985 / Riviera Hotel, Las Vegas, NevadaThe unit to be described differs from present tunable discriminator products chiefly because it is designed to simultaneously demodulate all the subcarrier channels in a frequency division multiplex. In addition the demodulated output data is presented in digital format that is compatible for direct computer entry. The discriminator implementation techniques will be discussed at the block diagram level. Particular emphasis will be given to the use of “Finite Impulse Response” filters and also to the internal tape speed compensation process.
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Qishan, Zhang, Zhang Wenguan i Mu Zhiying. "THE APPLICATION OF ORTHOGONAL FUNCTION TO MULTIPLEX". International Foundation for Telemetering, 1985. http://hdl.handle.net/10150/615706.
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International Telemetering Conference Proceedings / October 28-31, 1985 / Riviera Hotel, Las Vegas, NevadaA unified approach to the multiplex is proposed in terms of the orthogonal functions. It is called quadrature division multiplex, or Q D M in short. The orthogonal function is essential to the multiplex. Except these functions mentioned above, there are other orthogonal functions which are suitable for engineering practice. The orthogonality of the functions is used for the division of signals. A block diagram of Q D M is briefly described. The perfomances of the Q D M system are analysed. The speciality of the Q D M is simple and good. A model of the Q D M is built in our laboratory. A New type of bridge function is presented. It is called copy-shift bridge functions, which may be very useful for multiplex.
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Young, Mitchell Patrick. "A 3-D Multiplex Paper-microfluidic Platform". DigitalCommons@CalPoly, 2016. https://digitalcommons.calpoly.edu/theses/1680.
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3-D paper-based microfluidic devices (micoPADs) are small and portable devices made out of paper that offer a promising platform for diagnostic applications outside of a laboratory. These devices are easy to use, low cost, require no power source, and capable of detecting multiple targets simultaneously. The work in this thesis demonstrated the ability of a 3-D paper-microfluidic platform to simultaneously detect 5 targets. Rubber cord stock was used in conjunction with an acrylic housing unit to apply pressure along the edge of the channel. The indirect pressure application was successful in promoting vertical fluid flow between layers. Average channel development times were recorded between 110 seconds and 150 seconds.
The implementation of the 3-D paper-microfluidic platform as a diagnostic device was validated with a colorimetric glucose assay. In a novel application, reagents were deposited onto the 3-D platform via a glucose reagent pencil created by Martinez et al. A visual signal was observed for the successful detection of glucose at a concentration of 1.2 mM. These results offer promise for future work in combing new reagent deposition techniques with a multi-layer paper-microfluidic platform. Overall, this research made advancements in the design of a paper-microfluidic platform capable of the simultaneous detection of 5 targets.
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Novoa, Del Toro Elva Maria. "Detecting active modules in multiplex biological networks". Thesis, Aix-Marseille, 2020. http://theses.univ-amu.fr.lama.univ-amu.fr/200514_NOVOADELTORO_173hc776k263go601dzf_TH(1).pdf.
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L'expression des gènes est régulée dans le temps, les types de cellules et les conditions. Nous avons de nos jours accès à des technologies nous permettant de mesurer l'expression des gènes. Nous pouvons donc calculer les différences d'expression génique entre patients et témoins, et identifier ainsi les gènes dont l'expression est dérégulées. Nous pouvons aussi essayer de trouver un enrichissement des fonctions cellulaires à partir de la liste des gènes dérégulés. Dans ce contexte, j'ai analysé les données d'expression transcriptomiques de patients ayant le syndrome progéria de Hutchinson-Gilford (HGPS) et aux témoins sains. Ces analyses ont conduit à l'identification ARNs candidats pour validation expérimentale.À l'intérieur des cellules les molécules n'agissent pas isolément, mais interagissent pour accomplir ses fonctions. Actuellement, nous disposons de techniques pour déchiffrer ces interactions à grande échelle. Les interactions peut être représenté comme des réseaux, où les noeuds représentent des molécules, et les arêtes représentent des relations physiques et/ou fonctionnelles. L'hypothèse principale de ma thèse est que des sous-réseaux denses et dérégulé correspondent aux processus cellulaires affectés chez les patients. J'ai intégré des données d'expression génique et des réseaux pour identifier de tels modules. J'ai développé MOGAMUN, un algorithme génétique multi-objectif qui recherche des modules actifs. MOGAMUN est le premier algorithme d'identification de modules actifs capable de considérer les réseaux multiplexes, i.e. réseaux composés de différentes couches d'interactions biologiquesGene expression is regulated in time, cell types and conditions. We have access to technologies allowing us to measure the gene expression. We can therefore calculate the differences in gene expression between patients and controls, thereby identifying deregulated genes. We can also try to find significant enrichment of one or more cellular functions from the list of deregulated genes. I analyzed transcriptomics data corresponding to Hutchinson-Gilford Progeria Syndrome (HGPS) patients and healthy controls. Our analyses led to the identification of candidate RNAs for experimental validation.Inside cells the molecules do not act isolated, but they interact to accomplish their functions. Nowadays, we have experimental techniques to decipher these interactions on a large scale. Biological interactions can be represented as networks, where the nodes represent molecules, and the edges represent physical and/or functional relationships. The main hypothesis I followed during my thesis is that dense subnetworks associated to an overall expression deregulation correspond to affected cellular processes in patients. I integrated gene expression data and networks to identify such modules. I developed MOGAMUN, a multi-objective genetic algorithm that seeks for active modules. MOGAMUN is the first active module identification algorithm able to consider multiplex networks, i.e. networks composed of different layers of biological interactions
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Schaich, Frank. "Wellenlängen-Zeit-Codierung für dichten Wellenlängen-Multiplex /". Aachen : Shaker, 2008. http://d-nb.info/990928616/04.
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Vörös, András. "The emergence of multiple status systems in adolescent communities : a multiplex network theory of group formation". Thesis, University of Oxford, 2015. https://ora.ox.ac.uk/objects/uuid:9590194f-84e9-4548-b1fe-cf2f64ffc329.
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How do informal groups emerge in adolescent communities? What distinguishes a group from just a set of students? Who will end up together in a group and who will be left out? Why are there more groups in some classrooms and fewer in others? What determines whether these groups overlap in their members or they are completely segregated, perhaps antagonistic? While a huge body of research in sociology and social psychology focuses on these questions, an integrated approach that is able to answer all of them is yet to be developed. Without realizing that these five issues are interrelated, we cannot hope to understand how groups influence individuals and how they shape our communities. This thesis proposes an integrative theory of informal group formation in communities. Based on the tradition of Social Network Analysis, it develops a framework in which interpersonal relations and reputations are formed through a process called informal status production. Groups emerge from this micro-process by the alignment of positive relations and agreement in peer-perceptions between actors. The main micro-mechanisms predicted by the theory are tested on a unique longitudinal network dataset from school classrooms. To this end, a new empirical procedure was developed, by which a few aggregated networks can be constructed from tens of relational items. This allows the analysis of rich network data with several relational dimensions. The empirical studies of multiplex network dynamics confirm that there are strong interdependencies between friendships and perceptions. Students who agree about their peers tend to become friends, but more so when they hold a minority opinion in the class. This contributes to group formation. Friends also influence each other's perceptions, but we manage to show that the presence of groups around them interferes with this process by moderating the influence of individual peers.
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Wang, Yu. "Development of a multiplex fluorescent immunoassay for the simultaneous detection of serum antibodies to multiple swine pathogens". Thesis, Kansas State University, 2013. http://hdl.handle.net/2097/15704.
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Master of ScienceDepartment of Diagnostic Medicine and Pathobiology
Raymond R. R. Rowland
Three economically important swine diseases: Porcine Reproductive and Respiratory Syndrome (PRRS), Porcine Circovirus Associated Disease (PCVAD) and Swine influenza cost the US swine industry more than a billion dollars each year. This study developed a fluorescent microsphere immunoassay (FMIA) to simultaneously detect antibodies to the causative pathogens: PRRSV, porcine circovirus (PCV2) and swine influenza virus (SIV). The results showed that the multiplex assay possessed the predicted specificities. In the case of PRRSV NA, the assay displayed higher sensitivity when compared to a commercially available ELISA. The assay was employed to measure both IgG and IgM responses. The FMIA was found to possess several advantages over standard ELISA which include reduced sample volume, time and cost and provides a new tool for veterinary diagnostics. The FMIA was applied for swine disease surveillance in Hawaiian and Texan feral swine populations. The antibodies against PCV2 showed the highest prevalence among these three pathogens in both Hawaii and Texas. Hence we consider PCV2 as the most prevalent pathogen in Hawaiian and Texan feral pigs and this pathogen poses the greatest threat to commercial pigs. SIV seroprevelance increased from 2007 to 2010 in Hawaii State, suggesting an increasing risk for commercial pigs. Moreover, yearly surveillance in Texas State shows growth in seropositive response to all pathogens, particularly PCV2. The development of FMIA for detection of antibodies to multiple swine pathogens in serum samples offers an important alternative for swine disease surveillance in commercial and feral herds.
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Kirchner, Piero. "Multiplex-System für digitale Datenströme in multimedialen Anwendungen /". Hamburg : New-Business-Verl, 2010. http://d-nb.info/1000027724/04.
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Stella, Massimo. "Network structure and dynamics of empirical multiplex systems". Thesis, University of Southampton, 2017. https://eprints.soton.ac.uk/419488/.
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Interactions are important, since they can influence and shape a variety of real-world networked systems. Interactions can have a multi-relational nature, i.e. being of different type. Different layers of interactions can look very different from each other, motivating the need for developing multi-layer network models and metrics in network science. This thesis aims at developing novel multiplex frameworks for the quantitative investigation of two real-world systems: (i) the web of relationships between words in the human mind and (ii) the ecological interactions among animals in ecosystems. Despite being different in nature, both the mental lexicon of words and ecosystems can be represented as a multiplex network, where nodes represent distinct entities (e.g. words or animal groups) interacting on different layers in different ways (e.g. words being semantically and/or phonologically similar; animal species eating or parasitising each other). In both the considered systems, interactions crucially determine function and dynamics of a variety of processes. In the mental lexicon, individual interactions have been shown to influence both language acquisition and usage. In Part I of this thesis, I show that the structure of the phonological layer reflects constraints related to language use. I proceed by introducing the framework of multiplex lexical networks for quantifying, for the first time, the influence that phonology and semantics combined can have on (i) word acquisition of toddlers and (ii) word processing of adults. Results highlight phenomena that are not observable in single-layer networks. In toddlers word learning strategies based on the whole multiplex structure match empirical word learning significantly better than strategies based on individual layers, indicating that multiplexity is important for early word acquisition. At later ages the multiplex structure evolves by displaying an early, explosive emergence of a multiplex network core of words, which facilitates mental navigation and increases robustness against cognitive impairments. The second part of this thesis focuses on ecosystems, where interactions encapsulated in food webs or host-parasite networks greatly influence species extinction. In Part II of this thesis, I introduce the framework of ecological multiplex or “ecomultiplex” networks for combining predator-prey and host-parasite contact interactions as two layers of a network representing trophic links in a given ecosystem. I show that host-parasite interactions can dramatically increase the susceptibility of ecosystems to a parasite pandemic compared to models based on single-layer trophic networks only. Results of the ecomultiplex model are tested against empirical findings from field work in Brazilian ecosystems, finding agreement between my theoretical results and empirical data. Furthermore, by considering the multi-relational nature of trophic interactions, I quantitatively show that generalist top predators might accelerate parasite spread rather than hampering it, thus providing a theoretical explanation to recent empirical findings. Both in the mental lexicon and ecosystems, multiplexity influences structure, dynamics and function in ways not yet accounted for in the literature. This thesis aims to fill this gap by suggesting multiplex frameworks suitable for quantitative testing of empirical conjectures, while opening new modelling challenges at the interface of physics, network science and other disciplines.
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Cambronero, Christoffer. "Multiplex protein data and how to handle it". Thesis, Uppsala universitet, Statistiska institutionen, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-297390.
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Objective: To nd which methods and techniques that work well when working with multiplex proteindata. A short analysis of a dataset to show the methods is done at the end of the thesis. Material and methods: A study containing 171 individuals is used to test if there are any dierencesin protein values based on obesity, sugar level and type of obesity(5 subgroups). To investigate this OlinksProseek multiplex is used to generate 92 dierent protein values for each individual in the dataset. MainlyWelch's t-test and ANOVA are used as the comparison method for multiplex protein data. Since multipletests are performed the p-values are adjusted to avoid multiple type I error. Dierent ways to visualize thedata are also presented. Results: When comparing the obese and non-obese groups 25 proteins are shown to be signicantly dierent(p-value <0:05). Low and high sugar have no proteins that are signicantly dierent(p-value <0:05) betweenthe two groups and for the dierent types of obesity 18 proteins are signicant dierent(p-value <0:05). Conclusion: Both the tests for obesity and types of obesity show dierences between the groups while thesugar level do not have any proteins that signicantly dier between the groups. However when performinga multivariate techniques of low and high sugar the two can be separated indicating some dierences inprotein values.
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Ramos, Elisabete Manuela de Sousa. "Sistemas Multiplex para a deteção e caracterização molecular de infeções". Master's thesis, [s.n.], 2012. http://hdl.handle.net/10284/3575.
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Trabalho apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências FarmacêuticasAtualmente existe uma enorme preocupação em estudar os microrganismos, em particular os patogénicos para o Homem, uma vez que a sua presença no organismo constitui um risco potencial para a saúde Humana. Por este motivo, nos últimos 10-20 anos, temos assistido a uma evolução significativa nas técnicas disponíveis para a identificação laboratorial dos agentes responsáveis por patologias de base infeciosa. Os novos métodos, especialmente os baseados na tecnologia dos ácidos nucleicos, apresentam enormes vantagens em termos de sensibilidade, especificidade e grande rapidez na obtenção de resultados. Por este motivo, têm sido cada vez mais utilizados na deteção e identificação de microrganismos com interesse diagnóstico, prognóstico e de orientação do tratamento. Diversas tecnologias moleculares são utilizadas em laboratório para detetar agentes patogénicos. As mais utilizadas dependem de plataformas de PCR, e mais recentemente de PCR-em-tempo-real, disponibilizando sensibilidade, especificidade e rapidez sem precedentes na história da microbiologia e virologia clínica. No entanto a elevada especificidade destas técnicas podem também constituir uma desvantagem em situações clínicas que possam ter etiologias associadas a um largo espetro de microrganismos. Por este motivo têm sido desenvolvidas estratégias moleculares para detetar simultaneamente múltiplos agentes: os designados Sistemas Multiplex. O primeiro destes sistemas a ser desenvolvido foi o PCR Multiplex o qual permite detetar e amplificar simultaneamente mais do que um organismo identificando assim o agente etiológico. Mais recentemente têm sido desenvolvidos outros métodos multiplex com o mesmo fim: Técnicas baseadas em Microchips (de hibridização ou de amplificação as quais permitem a deteção e identificação de até milhares de sequências e consequentemente centenas a milhares de agentes patogénicos em simultâneo) bem comos sistemas baseados na tecnologia X-Map (fusão da citometria de fluxo e hibridização em solução com partículas fluorescentes). Do mesmo modo, os sistemas baseados na amplificação e sequenciação do gene rRNA 16S permitem a deteção e identificação de praticamente qualquer bactéria patogénica, sem necessitar de qualquer suspeita prévia. Finalmente, o advento das técnicas de sequenciação de 2ª e 3ª geração permitiu o desenvolvimento de uma nova área científica: a Metagenómica, a qual procura caracterizar simultaneamente e completamente a composição microbiológica de ecossistemas complexos, sejam eles humanos (pele, urina, fezes, boca, etc) ou ambientais (ar, águas residuais, solo, etc). Atualmente existe uma enorme preocupação em estudar os microrganismos, em particular os patogénicos para o Homem, uma vez que a sua presença no organismo constitui um risco potencial para a saúde Humana. Por este motivo, nos últimos 10-20 anos, temos assistido a uma evolução significativa nas técnicas disponíveis para a identificação laboratorial dos agentes responsáveis por patologias de base infeciosa. Os novos métodos, especialmente os baseados na tecnologia dos ácidos nucleicos, apresentam enormes vantagens em termos de sensibilidade, especificidade e grande rapidez na obtenção de resultados. Por este motivo, têm sido cada vez mais utilizados na deteção e identificação de microrganismos com interesse diagnóstico, prognóstico e de orientação do tratamento. Diversas tecnologias moleculares são utilizadas em laboratório para detetar agentes patogénicos. As mais utilizadas dependem de plataformas de PCR, e mais recentemente de PCR-em-tempo-real, disponibilizando sensibilidade, especificidade e rapidez sem precedentes na história da microbiologia e virologia clínica. No entanto a elevada especificidade destas técnicas podem também constituir uma desvantagem em situações clínicas que possam ter etiologias associadas a um largo espetro de microrganismos. Por este motivo têm sido desenvolvidas estratégias moleculares para detetar simultaneamente múltiplos agentes: os designados Sistemas Multiplex. O primeiro destes sistemas a ser desenvolvido foi o PCR Multiplex o qual permite detetar e amplificar simultaneamente mais do que um organismo identificando assim o agente etiológico. Mais recentemente têm sido desenvolvidos outros métodos multiplex com o mesmo fim: Técnicas baseadas em Microchips (de hibridização ou de amplificação as quais permitem a deteção e identificação de até milhares de sequências e consequentemente centenas a milhares de agentes patogénicos em simultâneo) bem comos sistemas baseados na tecnologia X-Map (fusão da citometria de fluxo e hibridização em solução com partículas fluorescentes). Do mesmo modo, os sistemas baseados na amplificação e sequenciação do gene rRNA 16S permitem a deteção e identificação de praticamente qualquer bactéria patogénica, sem necessitar de qualquer suspeita prévia. Finalmente, o advento das técnicas de sequenciação de 2ª e 3ª geração permitiu o desenvolvimento de uma nova área científica: a Metagenómica, a qual procura caracterizar simultaneamente e completamente a composição microbiológica de ecossistemas complexos, sejam eles humanos (pele, urina, fezes, boca, etc) ou ambientais (ar, águas residuais, solo, etc).
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Albuquerque, Renata Chaves. "Avaliação de um ensaio utilizando-se MULTIPLEX-PCR para a detecção de meningites por diferentes agentes bacterianos". Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/9/9136/tde-24012017-102958/.
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No presente trabalho, foi realizada uma MULTIPLEX-PCR para a detecção do DNA bacteriano de S. agalactiae, S.pneumoniae, N. meningitidis, H. influenzae e outros possíveis agentes etiológicos bacterianos das meningites. Este ensaio combina cinco diferentes iniciadores que detectam simultaneamente o gene crtA de N. meningitidis, o gene p6 de H. influenzae, o gene fbsA de S. agalactiae, o gene lytA de S. pneumoniae e o gene universal 16S rDNA para identificar a presença de agente bacteriano. Foram analisadas 447 amostras de LCR, o ensaio detectou 27 amostras positivas para cultura bacteriana e 13 amostras com resultado de cultura negativa. Estas amostras com cultura negativa apresentavam alterações bioquímicas, hematológicas, imunológicas ou microbiológicas (bacterioscopia) sugestivas de meningite, estes dados auxiliaram na análise dos resultados do MULTIPLEX-PCR. Este ensaio não apresentou reações inespecíficas com fungos, vírus e com outros agentes bacterianos testados (amplificando somente o gene 16S rDNA). A MULTIPLEX-PCR é um ensaio rápido, confiável, de fácil execução e facilmente implementável para a confirmação de meningite bacteriana. E este método pode auxiliar no diagnóstico de meningite com cultura de LCR negativa, particularmente para pacientes que previamente iniciaram antibioticoterapia e no diagnostico diferencial de meningite bacteriana ou viral.In this work, a MULTIPLEX-PCR has conducted for the bacterial DNA detection of S. agalactiae, S.pneumoniae, N. meningitidis, H. influenza and other possible etiologic bacterial meningitis agents. This test combines five different primers that detect simultaneously the crtA gene of N. meningitides, the p6 gene of H. influenza, the fbsA gene of S. agalactiae, the lytA gene of S. pneumoniae and the 16S rDNA universal gene to identify the presence of bacterial agent. From the 447 samples of CSF that were analyzed, the test detected 27 positive samples for bacterial culture and 13 samples with the result of negative culture. These negative culture samples presented biochemical changes, hematological, immunological or microbiological (bacterioscopy) suggestive of meningitis, these data helped in the analysis of the MULTIPLEX-PCR results. This test showed no nonspecific reactions with fungi, viruses and other bacterial agents tested (only amplifying the gene 16S rDNA). The MULTIPLEX-PCR test is a fast, reliable, easy to implement and easily implementable for of bacterial meningitis confirmation. And this method can aid in the meningitis diagnosis with negative culture of CSF, particularly for patients who previously started antibiotic therapy and in the differential diagnosis of bacterial or viral meningitis.
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Migoyan, Ara-Shant. "Development and evaluation of procedures and methods for Proseek Multiplex". Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-221287.
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Contemporary proximity extension assays (PEAs) are used for qualitative proteinquantifications in serological samples, with possibilities for scaling assays in multiplex. Medical research can however benefit from robust immunoassays functional for assessingprotein levels in other types of biospecimens. Formalin-fixed paraffin embedded (FFPE)tissues have long been used for morphological studies. The proteome encapsulated byextensive cross-linking from formalin fixation has however impeded the development ofproteomic analysis from the vast biorepositories FFPE-tissues constitute. In this study, Ipresent a proof of concept for assessing FFPE-samples in multiplex format through PEA.Furthermore, a homogenization and protein extraction protocol for assessing fresh-frozentissue with PEA is presented, together with a novel sample buffer for which remarkable risesin protein detection can be seen in several protein assays. Together, these findings extend theapplication area of PEA to tissues together with improved quantification characteristics.
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Mir, de Simón Bernat. "Multiplex optical detection system for prosthetic joint infection diagnosis". Doctoral thesis, Universitat Autònoma de Barcelona, 2016. http://hdl.handle.net/10803/400573.
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La infección de prótesis articular (IPA) es una patología de difícil manejo terapéutico que produce tanto morbilidad física como psicológica en los pacientes afectados y, además, está asociada con un impacto económico sustancial sobre el sistema sanitario. Su incidencia oscila entre 0.8 y 2.4% dependiendo de la articulación afectada, tales como la muñeca, el hombro, el codo, la rodilla o la cadera. A pesar de la baja incidencia de la IPA, el coste económico que se desprende ella es considerable (566M$ en 2009 para el sistema sanitario Americano). Por otro lado, un reconocimiento y manejo adecuados son críticos para preservar y restablecer la correcta funcionalidad de la articulación así como para evitar la morbilidad. Sin embargo, las infecciones crónicas tardías son difíciles de diagnosticar y normalmente están causadas por microorganismos que crecen en biopelículas. En ese estado, los microbios están protegidos contra agentes antimicrobianos así como de la respuesta inmune del huésped. Por lo tanto, una combinación de procedimientos complejos y de larga duración son necesarios para el diagnóstico de la IPA. Eso incluye, hallazgos clínicos, resultados de laboratorio de sangre periférica o líquido sinovial, datos microbiológicos, evaluación histológica del tejido periprotésico, inspección intraoperatoria y, en algunos casos, resultados radiográficos. Hoy en día, la técnica más usada para la identificación del agente infeccioso sigue siendo el cultivo celular. Desgraciadamente, esta técnica requiere entre 4 y 14 días para proporcionar un diagnóstico concluyente y dispone de una baja sensibilidad lo cual afecta al pronóstico de la infección.
En este trabajo se desarrolló un dispositivo basado en partículas plasmónicas modificadas con anticuerpos o aptámeros y Espectroscopia Raman Intensificada por Superfície (SERS) para la mejora del diagnóstico de la IPA. Dicho dispositivo fue capaz de identificar, con una sensibilidad por debajo de una unidad formadora de colonia, diferentes bacterias en un fluido biológico. Se obtuvo una detección simultanea y específica para cuatro agentes bacterianos (S. Aureus, E. Coli, S. Agalactiae y P. Aeruginosa). Para cada patógeno a identificar, se prepararon nanopartículas altamente eficientes en SERS con una combinación única de marcador Raman y anticuerpo/aptámero. Las partículas produjeron una señal Raman relativamente débil cuando estaban dispersas en el fluido. No obstante, cuando uno o más de los patógenos a identificar estaba presente, se produjo una acumulación de las nanopartículas asociadas sobre la membrana del microorganismo. Debido a ello, se formaron un numero considerable de “puntos calientes” en los que la señal Raman fue intensificada varios órdenes de magnitud (103) comparado con la señal que producían el mismo número de partículas dispersas en el fluido. De esta manera, la señal SERS fue lo suficientemente intensa para registrar cualquier agente bacteriano recubierto de nanopartículas que pasaba por el sistema de microfluidos. Este método fue capaz de monitorizar mililitros de diferentes fluidos biológicos (suero, sangre, orina y líquido pleural o ascítico) con un caudal de 0.1 mL/min. El estudio solo se realizó con cuatro patógenos pero el método de detección puede ser usado para la identificación simultánea de muchos otros microorganismos gracias al gran número de marcadores raman y anticuerpos/aptámeros selectivos disponibles. Por otro lado, pese a que el dispositivo aun no es capaz de discernir entre microorganismos resistentes o no resistentes, ofrece un diagnóstico preciso y en muy poco tiempo (menos de 15 minutos) comparado con los métodos de identificación comunes más usados (cultivo celular, reacción en cadena de polimerasa, ensayo por inmunoadsorción ligado a enzimas y analizador de tiempo de vuelo/desorción mediante láser asistida por matriz). Además, es capaz de monitorizar el efecto del tratamiento durante la evolución de la infección, aportando una herramienta muy útil para el correcto tratamiento de la IPA.Prosthetic Joint infection (PJI) is a disease of difficult therapeutic management that causes significant physical and psychological morbidity on patients and is associated with substantial financial burden on the healthcare system. Its incidence oscillates between 0.8-2.4% depending on the affected joint such as wrist, shoulder, elbow, hip and knee. In spite of the relatively low incidence of PJI, the financial burden remains enormous (i.e., $566 million in 2009 alone to the American health care system). Appropriate recognition and management are critical to preserving or restoring adequate function and preventing morbidity. However, late chronic infections are challenging to predict and typically caused by microorganisms that grow in biofilms. In that state, microbes are protected from antimicrobial agents and host immune responses. Therefore, a complex and time-consuming combination of procedures is required for its diagnosis. They include clinical findings, laboratory results from peripheral blood and synovial fluid, microbiological data, histological evaluation of periprosthetic tissue, intraoperative inspection, and, in some cases, radiographic results. Nowadays, microbial culture remains the most widespread technique for identifying the infectious agent, but unfortunately it requires 4-14 days to provide a conclusive diagnosis and has low sensitivity, affecting the prognosis of the infection. In order to develop a fast, accurate, and inexpensive method to improve the diagnosis of PJI, a device based on antibody/aptamer-modified plasmonic nanoparticles and Surface Enhanced Raman Scattering (SERS) spectroscopy was developed in this work. This device for infection diagnosis recognized, with sensitivity down to the single colony-forming unit, the presence of bacterial pathogens in biological fluids. The concept was confirmed with four bacterial agents (S. aureus, E. coli, S. agalactiae and P. aeruginosa). For each targeted pathogen receptor, high SERS efficient nanoparticles with a unique combination of a Raman label and an antibody/aptamer were prepared. The nanoparticles produced a relatively weak Raman signal when they are dispersed in a fluid. In contrast, the presence of one of the targeted pathogens triggered the accumulation of its partner nanoparticles on the antigen-carrying membrane of the microorganism, rapidly reaching full random coverage. Multiple gaps between nanoparticles are then formed that act as optical hotspots in which Raman scattering is enhanced by several orders of magnitude (103) relative to the same number of non-interacting nanoparticles. The resulting surface-enhanced Raman scattering signal is sufficiently intense as to record pathogen-specific inelastic light spectra from the nanoparticle-covered bacteria by driving the sample through a millifluidic channel. The method screened milliliters of different biological fluids (i.e., serum, blood, urine and pleural and ascites fluids) at a flow rate close to 0.1 mL/min. Although the concept was only confirmed with four bacterial agents, the detection method can be expanded to simultaneously identify more pathogens by using the large number of available spectral codes and selective antibodies/aptamers. On the other hand, this device is not yet capable of discerning directly in between resistant and non-resistant microorganisms, but it offers an accurate diagnosis in very short time (less than 15 minutes) as compared with all other currently available methods (i.e., cell culturing, matrix-assisted laser desorption/ionization-time-of-flight, polymerase chain reaction and enzyme-linked immunosorbent assay), and it can monitor the effect of the treatment on the time evolution of the infection.
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Granell, Martorell Clara. "From community structure to the physics of multiplex networks". Doctoral thesis, Universitat Rovira i Virgili, 2016. http://hdl.handle.net/10803/365314.
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Els sistemes complexes són sistemes que estan formats per un gran nombre d'unitats individuals, les quals interactuen entre elles, generalment de manera no-lineal, donant lloc a xarxes complexes d'interaccions. Sistemes biològics, socials i econòmics, entre d'altres, són sovint sistemes d'aquest tipus, el caràcter complex dels quals és mereixedor d'estudi. En els últims anys, un gran nombre de publicacions científiques han recolzat la conveniència d'analitzar aquest tipus de sistemes a través d'un conjunt d'eines i mètodes especialment dissenyats per la descripció de sistemes complexes. L'objectiu d'aquesta tesi és estudiar les xarxes complexes, i en particular, les xarxes multiplex. Estudiarem la caracterització topològica de les xarxes fent servir eines d'anàlisi de comunitats, posant especial èmfasi en els mètodes de multi-resolució, que són capaços d'informar-nos de tota la mesoescala. També ens ocuparem de caracteritzar processos dinàmics en xarxes multiplex, en el cas especial on dos processos epidèmics competeixen l'un contra l'altre sobre la mateixa estructura. Finalment, demostrarem l'aplicabilitat de les eines de xarxes complexes en un treball experimental dedicat a caracteritzar l'estructure funcional de cultius de neurones clusteritzades.
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Ustunel, Eser Kwon Hyuck M. "Time division duplex-wideband code division multiplex (TDD-WCDMA)". Diss., Click here for available full-text of this thesis, 2006. http://library.wichita.edu/digitallibrary/etd/2006/t029.pdf.
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Thesis (M.S.)--Wichita State University, Electrical and Computer Engineering."May 2006." Title from PDF title page (viewed on October 19, 2006). Thesis adviser: Hyuck M. Kwon. Includes bibliographic references (leaves 40-42).
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Mewis, Guido. "Entwicklung einer Multiplex-PCR zum Nachweis Enteropathogener Escherichia coli". [S.l.] : [s.n.], 2004. http://www.diss.fu-berlin.de/2004/93/index.html.
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Renoust, Benjamin. "Analysis and Visualisation of Edge Entanglement in Multiplex Networks". Phd thesis, Université Sciences et Technologies - Bordeaux I, 2013. http://tel.archives-ouvertes.fr/tel-00942358.
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When it comes to comprehension of complex phenomena, humans need to understand what interactions lie within them.These interactions are often captured with complex networks. However, the interaction pluralism is often shallowed by traditional network models. We propose a new way to look at these phenomena through the lens of multiplex networks, in which catalysts are drivers of the interaction through substrates. To study the entanglement of a multiplex network is to study how edges intertwine, in other words, how catalysts interact. Our entanglement analysis results in a full set of new objects which completes traditional network approaches: the entanglement homogeneity and intensity of the multiplex network, and the catalyst interaction network, with for each catalyst, an entanglement index. These objects are very suitable for embedment in a visual analytics framework, to enable comprehension of a complex structure. We thus propose of visual setting with coordinated multiple views. We take advantage of mental mapping and visual linking to present simultaneous information of a multiplex network at three different levels of abstraction. We complete brushing and linking with a leapfrog interaction that mimics the back-and-forth process involved in users' comprehension. The method is validated and enriched through multiple applications including assessing group cohesion in document collections, and identification of particular associations in social networks.
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Kownarumit, Supaporn. "Multiplex screening using enzyme inhibition, fluorescence detection and chemometrics". Thesis, Loughborough University, 2006. https://dspace.lboro.ac.uk/2134/12308.
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Fluorescence enzyme inhibition assays have been established for a number of years as valuable methods of analysis in clinical chemistry and other fields. Those in common use are normally single analyte assays. However, in many cases (e.g. drug screening) dual or multiplex assays would be much more valuable, with the advantages of increased information content, saving in time and costs, and the elimination of some sources of sampling variance. This project has investigated single and dual screening assays of enzyme inhibitors, namely 3-nitrophenylboronic acid (3-NPBA), phenylethyl /3-0- thiogalactopyranoside (PETG) and sodium vanadate (VI), using flow injection fluorescence spectroscopy and chemometric methods to resolve strongly overlapping fluorescence spectra. The single and dual screening assays have been based on flow injection analysis methodology, with immobilised enzymes on solid phase reactors to investigate the enzyme inhibitors. The assays were rapid, allowing around 15-25 measurements to be made per hour. The inhibitions of alkaline protease, alkaline phosphatase and /3-galactosidase with their inhibitors at flg/ml levels were achieved. An alternative approach to these dual assays has been investigated by the use of multivariate techniques. Such techniques allow accurate and reliable results to be obtained even from spectra that contain extremely overlapping signals. Moreover, preliminary investigation of three fluorophores which gave strongly overlapping spectra, using flow injection fluorescence spectroscopy and partial least squares (PLS-1) model has been successful. By combination of this flow injection fluorescence spectroscopy with the use of chemometrics, many applications can be envisaged in biochemical, clinical, and pharmaceutical industries. With the findings of this research the system described here can be developed for use in high throughput screening of candidate drug molecules and many screening processes throughout the different industries.
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Finski, Alexei. "Multiplex Analytical Measurements in Single Cells of Solid Tissues". Thesis, Harvard University, 2012. http://dissertations.umi.com/gsas.harvard:10652.
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Most human organs consist of solid tissues. Most human disorders occur in solid tissues. No two cells are equivalent in native solid tissues as single cells widely vary in their biochemistry, specialization, and location. Yet, the fundamental limitations of solid-tissue processing and analysis have made it challenging to access the richness of molecular information in single cells of animal and human solid tissues. We have eliminated a set of limitations of solid-tissue processing and analysis. In this thesis, I first describe a new method for sampling single cells in live solid tissues. This method preserves the molecules of all molecular classes and thus fulfills the precondition for multiplexing within and across molecular classes. I then describe new analytical methods and strategies for massively multiplex analysis within and across molecular classes in each sampled single cell. Standard curves, the basis of analytical methods, can be constructed in all measurements and signals can be mapped to the corresponding quantities. Proof of principle experiments are presented. These methods will enable the quantification of the molecular mechanism of each sampled single cell in solid tissues by analytically measuring tens of proteins, transcripts and/or metabolites at once. By performing these measurements in human solid-tissue biopsies, we will be able to define a new category of diagnostic tests, to personalize single-cell pharmacology and to rapidly identify mechanistic biomarkers and drug targets.
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Schaich, Frank [Verfasser]. "Wellenlängen-Zeit-Codierung für dichten Wellenlängen-Multiplex / Frank Schaich". Aachen : Shaker, 2008. http://d-nb.info/1161305734/34.
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Haidar, Ibrahim. "Digital code division multiplex techniques for data transmission applications". Thesis, Aston University, 1985. http://publications.aston.ac.uk/8052/.
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Way, Jaw-Shiow Chu. "Specific detection of Salmonella by multiplex polymerase chain reaction". Diss., The University of Arizona, 1993. http://hdl.handle.net/10150/186207.
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This study evaluates the use of multiplex polymerase chain reaction (PCR) technology for detection of Salmonella species in pure cultures and also in environmental samples. Three sets of oligonucleotide primers were used in the PCR assay: PhoP primers specific to a 299 bp region in the phoP/phoQ loci of coliform pathogenic bacteria such as Salmonella, Shigella, E. coli and Citrobacter species, served as a presumptive indicator of enteric bacteria. Subsequently Hin and H-1i primers, which targeted a 236 bp region of the hin/H2 gene, and a 173 bp region of the H-1-i flagellar gene in Salmonella typhimurium respectively, were used for specific detection of salmonellae. Specificity and sensitivity of PCR amplified products were evaluated by ethidium bromide (EtBr) staining or gene probe analysis. Under optimal PCR conditions described in this study, Salmonella species can be specifically detected by these 3 primer sets. The sensitivity of detection in terms of whole cells was a 10⁻⁵ dilution (approximately 10³ cells) of boiled late log phase cultured cells (detection by EtBr) after 25 cycles of PCR and 10⁻⁸ dilution (approximately 10° cells) after a 50 cycle 'double PCR' protocol. A similar level of sensitivity was observed using a gene probe analysis (³²P end-labeling) to detect the PCR amplified products. The multiplex PCR products observed from known environmental isolates allowed identification of several isolates as Salmonella species. These results agreed with conventional analyses. The efficacy of this PCR technology for environmental applications was also evaluated by testing the primers on soil and environmental water samples. All unseeded soil samples showed no PCR amplified products when detected by EtBr staining after 25 and 50 cycles PCR. Only one pond surface water sample resulted in a positive PCR result. This water sample was also positive when tested by conventional methodologies. Seeded soil and well water samples all resulted in PCR products being observed from 25 or 50 cycles. These results indicate that the primers have the potential to specifically detect Salmonella species in environmental samples.
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Soar, Filho Ercy José. "Varius multiplex multiformis : epistemologia do self no pós-modernismo". reponame:Repositório Institucional da UFSC, 1997. https://repositorio.ufsc.br/handle/123456789/112200.
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Chauvat, Anne. "Analyse de réponses lymphocytaires T par Elispot et Fluorospot au cours du suivi de protocoles de vaccination : mise en évidence de l’effet du thiomersal présent dans la préparation vaccinale". Thesis, Paris Est, 2012. http://www.theses.fr/2012PEST0064.
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L'analyse de la réponse lymphocytaire T représente une exploration de plus en plus importante pour la compréhension de la physiopathologie de maladies et le suivi de protocoles d'immunothérapie. La mise en évidence de la production de cytokines par ces cellules permet d'identifier les sous populations de lymphocytes T (LT) et d'analyser leur fonctionnalité et leur état d'activation. L'Elispot est une technique déjà largement utilisée dans le suivi de la réponse vaccinale pour la détection de cellules sécrétrices de cytokines. Bien que la séroprotection soit le critère reconnu pour déterminer l'efficacité de la vaccination antigrippale, un rôle important de la réponse lymphocytaire T a été démontré dans le contrôle de l'infection. Nos travaux ont permis de mettre en évidence que l'utilisation de vaccins totaux pour le suivi de la réponse lymphocytaire T, par l'introduction de leurs excipients, peut induire un biais dans la détection des cytokines. Le thimérosal, conservateur utilisé dans certains vaccins, provoque in vitro lors de la détection d'interféron (IFN) par Elispot l'apparition de petits spots que nous avons associés a l'activation abortive précoce des LT. Leur taille permet cependant d'éliminer automatiquement du comptage ces spots non spécifiques. Contrairement à l'Elispot dont elle est dérivée, la technique Fluorospot permet l'analyse simultanée de plusieurs cytokines et est donc plus adaptée à la détection des profils de sécrétion et de la fréquence des LT polyfonctionnels, associés à un bon pronostic dans plusieurs pathologies. Nos résultats ont permis la validation technique de ce test pour la détection des couples IFN/interleukine (IL)-10 et IFN/IL-2. Grâce au développement au laboratoire d'une lignée produisant de l'IFN et l'IL-10, nous avons montré que le Fluorospot avait une sensibilité supérieure à la cytométrie intracellulaire pour la détection de l'IFN et de l'IL-10. Il était déjà connu que l'IL-10 était difficilement détectable par cytométrie, renforçant l'intérêt du Fluorospot pour détecter les cellules Tr1. L'utilisation du Fluorospot lors du suivi de la réponse vaccinale antigrippale a également permis de démontrer une équivalence de sensibilité avec l'Elispot. Au cours du suivi de ce protocole anti-grippal, nous avons montré à l'aide du Fluorospot que la réponse LT antigrippale était surtout caractérisée par une forte production d'IL-2 et que la détection isolée d'IFN a une faible sensibilité pour la mesure des réponses LT anti-grippales. Il s'agit de la première utilisation de ce test pour le suivi d'un protocole vaccinal démontrant sa robustesse et sa faisabilité dans un contexte clinique. Nos résultats offrent donc une validation complète de la technique Fluorospot, aussi bien du point de vue technique que clinique, ouvrant des perspectives d'utilisation pour le suivi de futurs protocolesThe detection of cellular response via its different T cell subpopulations was proved to be crucial to better understand mechanisms of pathologies and monitor protocols of immunotherapy. Unicellular cytokine measurement not only allows the assessment of their inherent role on immunological response, but also the detection of T lymphocytes (LT) secretion profile which gives information on subpopulations involved, their activation state and their functionality. Detection of the cytokine secreting cells frequency is achievable with Elispot. This test is widely used in clinical trials to monitor vaccine response to diseases such as influenza. Although seroprotection is the recognized parameter to assess anti-influenza vaccine efficiency, cellular response has been proved to play an important role in infection control. Our results highlighted that the use of the whole vaccine to monitor T cell response may induce bias in cytokine detection. Indeed, their excipients can be considered as contaminants, like Thimerosal which is used as a preservative in some vaccines. We demonstrated that it induced detection of small spots in interferon (IFN) Elispot. We proved that these spots were associated with in vitro early abortive T cell activation. However, their size enabled us to remove these unspecific spots from bona fide spots.Fluorospot test is suited for multiplex cytokine analysis. Thus it is more adapted than Elispot to detect cytokine secretion profiles and polyfunctional T cells frequency. It is worth noting that polyfunctionnal Tcells are associated with a better clinical outcome in several pathologies. Our results led to technical validation of this test for detection of two cytokines couples, IFN/interleukin (IL)-10 and IFN/IL-2. Fluorospot showed a better sensitivity than intracellular staining cytometry (ICS) in detection of IFN and IL-10 produced by a cell line transfected with the cDNA encoding these cytokines. Moreover, Fluorospot demonstrated a similar sensitivity than Elispot when used to monitor the immunological response to an anti-influenza vaccine. Furthermore, using this technique, we showed that anti-influenza T cell producing IL-2 were the dominant population of T cells. Isolated IFN producing T cells were less sensitive than the detection of IL-2 to detect specific T cell response against influenza. Therefore our results provided a full validation of Fluorospot test, for the technical part as well as for the clinical part. These conclusions open perspectives of using this method to monitor protocols in a near future
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Hélard, Jean-François. "Modulations codées en treillis associées à un multiplex de porteuses orthogonales en présence de canaux affectés de trajets multiples". Rennes 1, 1992. http://www.theses.fr/1992REN1S117.
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Šifta, Radim. "Kvalita služeb v optických přístupových sítích". Doctoral thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2015. http://www.nusl.cz/ntk/nusl-233668.
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The thesis focuses on the optical access networks and deals with the issue of transmission parameters of the physical layer, their negative impact on the quality of the services parameters and the possibilities of reducing their effect using the polarization multiplexing. The first and second chapter describe the theoretical background necessary for understanding the subsequent parts. The third chapter contains the practical part of the thesis. At first the optical routes in the Czech Republic in terms of polarization mode dispersion, which is together with insertion loss the main limiting parameter of high-speed optical networks, are evaluated. In the following part the measuring of the temperature changes effects on the polarization mode dispersion of the passive components and the effect of temperature changes to the polarization multiplexed signal are evaluated. To double the bandwidth and reduce the influence of polarization mode dispersion on the quality of services, the two simulation models of optical access networks using polarization multiplexing were carried out. These theoretical outputs were verified by the practical measurements in the laboratory and subsequently on the real optical route. Finally, a draft of the broadband optical access network based on wavelength and polarization multiplexing was designed based on obtained knowledge.
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Kibalo, Tom, i Ben Miles. "DEFINITION OF A FULLY COMPLIANT IRIG RECORDING SYSTEM FOR TELEMETRY". International Foundation for Telemetering, 1997. http://hdl.handle.net/10150/609774.
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International Telemetering Conference Proceedings / October 27-30, 1997 / Riviera Hotel and Convention Center, Las Vegas, NevadaAlliant Techsystems’ Advanced Technology Applications organization incorporates the latest IRIG standards for range equipment and operation. Over the past five years, our objective has been to assure interoperability among diverse data recording users while achieving technical excellence for our ADARIO(Analog Digital Adaptable Input Output) family of products. In this paper, we summarize 25 years of ADARIO development; technical challenges, risks and processes; as well as our five-year effort to modify and develop our recording system products to meet the evolutionary standards of technical excellence.
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Almeida, Isa Azevedo de. "Padronização da tecnica para identificação do Nanoplex Y-STR em amostras de sangue humano". [s.n.], 2003. http://repositorio.unicamp.br/jspui/handle/REPOSIP/290746.
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Orientador: Eduardo Daruge JuniorDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
Made available in DSpace on 2018-08-03T09:15:14Z (GMT). No. of bitstreams: 1 Almeida_IsaAzevedode_M.pdf: 4484053 bytes, checksum: e5137b49df6ad4e6c1204bd7f37d39a9 (MD5) Previous issue date: 2003
Resumo: Nas últimas décadas, as Ciências Forenses vêm evoluindo de maneira a esclarecer e auxiliar cada vez mais precisamente a justiça. O presente trabalho, através da utilização de 25 amostras de sangue de cadáveres do Instituto Médico Legal de Cuiabá-MT, teve como objetivos: estabelecer a forma de coleta do material biológico através da utilização de cartões FTA@; aprimorar a extração do DNA em amostras de sangue, utilizando a Resina Quelante CHELEX@; aprimorar a PCR utilizando os primers para os loei DYS 3851/11, DYS 3891/11, DYS 390, DYS 391, DYS 392, DYS 393, DYS 19 através da padronização do Nanoplex; comprovar a importância do estudo do cromossomo Y nos casos de investigação de paternidade e em casos forenses. Com o desenvolvimento do trabalho, pôde se observar que os métodos de coleta das amostras de sangue e extração do DNA foram satisfatórios, pois obteve-se uma quantidade de DNA suficiente para o seu estudo. O estudo do Nanoplex fez-se com sucesso, pois, por meio de sua análise em um seqüenciador automático (ASI PRISM DNA Sequencer 377), pôde se detectar os fragmentos, estipulando-se, assim, seus respectivos alelos
Abstract: In the last decades, the Forensic Sciences come evolving in way to clarify and to assist the justice. The present work through the use of 25 samples of corpse blood of the Legal Medical Institute of Cuiabá-MT had as objective: to establish the fonn of collection of the biological material through the use of cards FT A@; to improve the extraction of the DNA in samples of blood being used the Resin Quelante CHELEX@; to improve the PCR using primers for locus DYS 385 1/11, DYS 389 1/11, DYS 390, DYS 391, DYS 392, DYS 393, DYS 19 through the standardization of the Nanoplex; to prove the importance of Y chromosome study in the cases of patemity and in forensic cases. Wlth the development of the work it could be observed that the methods of collection of blood samples and extration of the DNA had been satisfactory, therefore got a enough amount of DNA for its study. The study of the Nanoplex one became successfully, therefore through its analysis in an automatic sequencer (ASI PRISM DNA Sequencer 377), it could be detected the fragments, stipulating its respective alleles
Mestrado
Odontologia Legal e Deontologia
Mestre em Odontologia Legal e Deontologia
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Del, Valle Mendoza Juana, i Universidad Peruana de Ciencias Aplicadas (UPC). "Diagnóstico de 14 virus respiratorios y 3 gérmenes atípicos en pacientes inmunodeprimidos mediante la técnica RT-PCR multiplex". Universidad Peruana de Ciencias Aplicadas (UPC), 2015. http://hdl.handle.net/10757/345682.
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Class, Volker. "Entwicklung und Struktur des deutschen Kinomarktes". [S.l. : s.n.], 2003. http://www.bsz-bw.de/cgi-bin/xvms.cgi?SWB11675626.
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Zandegiacomo, Cella Alice. "Multiplex network analysis with application to biological high-throughput data". Master's thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amslaurea.unibo.it/10495/.
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In questa tesi vengono studiate alcune caratteristiche dei network a multiplex; in particolare l'analisi verte sulla quantificazione delle differenze fra i layer del multiplex. Le dissimilarita sono valutate sia osservando le connessioni di singoli nodi in layer diversi, sia stimando le diverse partizioni dei layer. Sono quindi introdotte alcune importanti misure per la caratterizzazione dei multiplex, che vengono poi usate per la costruzione di metodi di community detection . La quantificazione delle differenze tra le partizioni di due layer viene stimata utilizzando una misura di mutua informazione.
Viene inoltre approfondito l'uso del test dell'ipergeometrica per la determinazione di nodi sovra-rappresentati in un layer, mostrando l'efficacia del test in funzione della similarita dei layer.
Questi metodi per la caratterizzazione delle proprieta dei network a multiplex vengono applicati a dati biologici reali. I dati utilizzati sono stati raccolti dallo studio DILGOM con l'obiettivo di determinare le implicazioni genetiche, trascrittomiche e metaboliche dell'obesita e della sindrome metabolica. Questi dati sono utilizzati dal progetto Mimomics per la determinazione di relazioni fra diverse omiche. Nella tesi sono analizzati i dati metabolici utilizzando un approccio a multiplex network per verificare la presenza di differenze fra le relazioni di composti sanguigni di persone obese e normopeso.
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Santos, Sílvia Regina dos. "Determinação de sorotipos capsulares de Streptococcus pneumoniae por Multiplex-PCR sequencial". Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/9/9136/tde-12092012-100134/.
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S. pneumoniae coloniza a nasofaringe e é um dos principais agente de otite média, pneumonia, bacteremia e meningite com altas taxas de morbidade e mortalidade. Estima-se que 1,6 milhões de pessoas morram de doença pneumocócica por ano, a maioria crianças menores de cinco anos de idade, principalmente em países em desenvolvimento. A cápsula polissarídica antifagocitária é o principal fator de virulência deste microrganismo e determina os 93 sorotipos conhecidos, sendo o alvo de vacinas pneumocócicas. No presente trabalho foi padronizada a tipagem molecular por Multiplex PCR de S. pneumoniae, que compreende 30 pares de iniciadores agrupados em seis reações sequenciais. Foram tipadas 270 cepas de pneumococo isoladas entre janeiro de 2005 a setembro de 2011, proveniente de líquor (13%), sangue (76%) e líquido pleural (11%) de 232 pacientes atendidos no Hospital Universitário da USP. Além disso, a caraterização dessas amostras quanto ao perfil de sensibilidade aos antimicrobianos e à diversidade foi realizada, segundo o CLSI 2011 e a genotipagem molecular pelas técnicas de Multilocus Sequencie Typing Scheme (MLST) e Pulsed Field Eletrophoresis Gel (PFGE), respectivamente. A tipagem por Multiplex PCR detectou 24 sorotipos/sorogrupos diferentes, que foram: 14 (22%), 5 (12%), 12F/A (11%), 6A/B/C (10%), 7F/A (5%), 1 (4%), 3 (4%), 10A (4%), 19A (4%), 18 A/B/C/F (3%), 4 (3%), 8 (3%), 23F (3%), 19F (3%) e outros (9%) (9V/A, 9N/L, 15A, 22F, 11A/D, 31, 38, 34, 16F, 17F e não tipável). Este método apresentou 100% de especificidade e 98% de sensibilidade para determinação de sorogrupos e 66% para sorotipos. Os sorotipos 14, 6B, 5 e 19F foram significativamente mais comuns em criança até dois anos, já entre adultos, os sorotipos 5 e 12F foram os predominantes. O perfil de sensibilidade em infecções não meníngeas foi de 99% de sensibilidade e 1% de resistência intermediária para penicilina e ceftriaxona. Para infecções meníngeas os resultados mostraram 73% de sensibilidade e 27% de resistência para penicilina e 88% de sensibilidade e 12% de resistência intermediária para ceftriaxona. A resistência aos beta-lactâmicos está ligada principalmente ao sorotipo 14 que foi o sorotipo mais isolado com 52 cepas e dessas foram realizados MLST e PFGE. No MLST encontramos 51 cepas pertencentes ao clone Spain9V-3 (ST 156) que é predominante na região sul e sudeste do Brasil e uma cepa com um tipo de sequência ainda não depositada. Pela técnica de PFGE foram detectados três clusters e quatro amostras não relacionadas, o cluster A foi predominante com 41(79%) cepas com 81,7% de similaridade entre elas. A técnica de Multiplex PCR demonstrou ser excelente ferramenta para a detecção dos sorotipos/sorogrupos de S. pneumoniae. Não foi detectada resistência plena à penicilina e ceftriaxona em infecções não meníngeas consolidando a importância do uso da penicilina no tratamento da doença pneumocócica não meníngea. Houve grande similaridade genética entre cepas de S. pneumoniae sorotipo 14.S.pneumoniae colonizes the nasopharynx and is a major agent of otitis media, pneumonia, bacteremia and meningitis with high morbidity and mortality. It is estimated that 1.6 million people die of pneumococcal disease every year, mostly children under five years old, mainly in developing countries. The antiphagocytic polissarídica capsule is the main virulence factor of this organism and determine the 93 serotypes known for being the target of pneumococcal vaccines. In the present study was standardized molecular typing by Multiplex PCR molecular typing, which comprises 30 primer pairs grouped into six sequential reactions. We performed antimicrobial susceptibility profile, according to the CLSI 2011 and the most frequent serotype was made by molecular genotyping techniques Multilocus Sequence Typing (MLST) and pulsed-field gel Eletrophoresis (PFGE). We studied 270 pneumococcal strains isolated from 2005 to September 2011, from CSF (13%), blood (76%) and pleural fluid (11%) of 232 patients attended at University Hospital of USP. Typing by Multiplex PCR detected 24 serotypes / serogroups different, which were: 14 (22%), 5 (12%), 12F / A (11%), 6A/B/C (10%), 7F / A (5 %), 1 (4%), 3 (4%), 10A (4%), 19A (4%), 18 A / B / C / F (3%), 4 (3%), 8 (3% ), 23F (3%), 19F (3%) and others (9%) (9V / A, 9N / L, 15A, 22F, 11A / D, 31, 38, 34, 16F, 17F and nontypable). This method showed 100% specificity and 98% sensitivity for the determination of 66% for serogroups and serotypes. Serotypes significantly more common in children under two years were: 14, 6B, 5 and 19F among adults serotypes 5 e12F were predominant. The sensitivity profile in non-meningeal infections was 99% sensitivity and 1% penicillin intermediate resistance to ceftriaxone. For meningeal infections the results showed 73% sensitivity and 27% resistance to penicillin and 88% sensitivity and 12% intermediate resistance to ceftriaxone. Resistance to beta-lactams is linked mainly to serotype 14 was the serotype most isolated, and of these 52 strains were performed MLST and PFGE. MLST found in 51 strains belonging to clone Spain9V-3 (ST 156) which is prevalent in south and southeastern Brazil and a strain with a type of sequence is not deposited. The technique of PFGE found three clusters and four non-related samples, cluster A predominated with 41 (79%) strains with 81.7% similarity between them. Multiplex PCR technique proved to be an excellent tool for the detection of serotypes/serogroups of S. pneumoniae. We did not detect full resistance to penicillin and ceftriaxone in non-meningeal infections showing the importance of use of penicillin in the treatment of pneumococcal non-meningeal disease. There was great genetic similarity among strains of S. pneumoniae serotype 14.
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Dannered, Peter, i Eric Åkerhielm. "Utlokaslisering till Indien : En fallstudie av Multiplex utlokalisering av ekonomitjänster". Thesis, Uppsala universitet, Företagsekonomiska institutionen, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-169646.
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På grund av en hårdnande konkurrens på världsmarknaden har företags investeringar utomlands ökat frekvent de senaste åren. Fler och fler företag flyttar sina ekonomitjänster till lågkostnadsländer för att uppnå kostnadsfördelar. Indien utgör idag motiv för denna utlokalisering av ekonomitjänster främst på grund av deras höga utbildningsnivå, goda engelskkunskaper samt låga löner. Genom en egen modell som bygger på dessa teorier presenteras fyra bakomliggande faktorer; krav, kultur, kunskap och kostnader vilka utgör hinder och vidare möjligheter för utlokalisering av ekonomitjänster. Vidare visas vilken typ av ekonomitjänster som lämpar sig bäst för utlokalisering till Indien. Via en fallstudie av det multinationella företaget Multiplex utlokalisering av ekonomitjänster till Indien nås djupare insikt i problemet. I en analys granskas problemen och förslag presenteras på hur Multiplex kan överkomma de nämnda hindren. Resultatet visar vilken typ av ekonomitjänster som lämpar sig bäst att flytta till Indien samt vilka tjänster som Multiplex kan utlokalisera i framtiden.
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Sundequist, Blomdahl Kristofer. "An evaluation of random-walk based clustering of multiplex networks". Thesis, Uppsala universitet, Institutionen för informationsteknologi, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-333027.
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A network, or a graph, is a mathematical construct used for modeling relationships between different entities. An extension of an ordinary network is a multiplex network. A multiplex network enables one to model different kinds of relationships between the same entities, or even to model how relationships between entities change over time. A common network analysis task is to find groups of nodes that are unusually tightly connected. This is called community detection, and is a form of clustering. The multiplex extension complicates both the notion of what a community is, and the process of finding them.This project focuses on a random-walk based local method that can be used to find communities centered around supplied seed nodes. An implementation of the methodis made which is used to evaluate its ability to detect communities in different kinds of multiplex networks.
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Stenberg, Johan. "Software Tools for Design of Reagents for Multiplex Genetic Analyses". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-6832.
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Ereku, Luck Tosan. "Design of microfluidic multiplex cartridge for point of care diagnostics". Thesis, Brunel University, 2017. http://bura.brunel.ac.uk/handle/2438/15331.
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A simple, but innovative microfluidic Lab-on-a-chip (LOC) device which is broadly applicable in point of care diagnostics of biological pathogens was designed, fabricated and assembled utilising explicit microfluidic techniques. The purpose of this design was to develop a cartridge with the capability to perform multiplex DNA amplification reactions on a single device. To achieve this outcome, conventional laboratory protocols for sample preparation; involving DNA extraction, purification and elution were miniaturized to suit this lab-on-a-chip device of 75mm X 50mm cross-sectional area. The extraction process was carried out in a uniquely designed microchamber embedded with chitosan membrane that binds DNA at pH 5.0 and elutes when a different solution at pH 9.0 flows through. Likewise, purification protocol that occurs in the designed waste reservoir is very significant in biomedical field because it is concerned with waste treatment and cartridge disposability, was performed with a super absorbent powder that converts liquid to a gel like substance. This powder is known as sodium polyacrylate, which is also they treated with anti-bacterial chemicals to prevent environmental contamination. Furthermore, this process also employed the use of a passive valve for a precise fluid handling operation involving flow regulation from extraction to waste reservoir. In order to achieve the intended multiplexing function a multiplexer was created to distribute flow simultaneously through a bifurcated network of channels connected to six similar amplification microchambers. Prior to fabrication, computational fluid dynamics (CFD) simulation was utilized at flowrates less than 10μL/s as the means to test the effectiveness of each design components and also to specifically deduct empirical values that can be analyzed to improve or understand the relationship between the fluid and geometrical constraints of the microfluidic modular elements. The device produced was a hybrid cartridge composed of PDMS and glass which is the most widely used materials microfluidics research due to their low cost and simplicity of fabrication by soft lithography technique. The choice of material also took into account the various physical and chemical properties advantages and disadvantages in their bio-medical applications. Such properties include but not limited to surface energy that determines the wetting fluid characteristics, biocompatibility, optical transparency. Subsequently, after a prototype cartridge was developed fluid flow experimentation using liquid coloured dye was used on the fully fabricated cartridge to test the efficacy of its microfluidic functionalities before expensive DNA amplification reagents were utilised at similar flowrates to the CFD simulations. This gave rise to comparison between similar and dissimilar flow Peculiarities in the microfluidic circuit of both experiments. The final experiment was performed with the aid of a recent molecular technique in DNA amplification known as of RPA kit (recombinase polymerase amplification reaction). It involved performing two main reaction experiments; first, was the positive experiment that bears the sample DNA and the latter, negative that served as the control without DNA. In the end, quantitative analysis of results was done using an agarose gel that showed 143 base pairs, for the positive samples, thus validating the amplification experiment.
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Wanfang, Zhang. "THE TIME DIVISION MULTIPLEX MEASURING SYSTEM FOR SINGLE-TRANSIENT SIGNALS". International Foundation for Telemetering, 1993. http://hdl.handle.net/10150/608868.
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International Telemetering Conference Proceedings / October 25-28, 1993 / Riviera Hotel and Convention Center, Las Vegas, NevadaIn order to reduce the measuring channels for the single-transient signals, the author propose the time division multiplex technique and introduce the method of SAW delay line in this paper. That used method of SAW tap-delay line in this system is different from previous methods consists in making traditional method, which is one-path signal input different delayed multi- path signals output, alter new method, which is simultaneous multi-path signal inputs that are respectively delayed and one-path signal serial output.
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Armbruster, Katrin. "Entwicklung einer Multiplex-TaqMan-PCR zur Diagnostik der viralen Enzephalitis". [S.l. : s.n.], 2006. http://nbn-resolving.de/urn:nbn:de:bsz:25-opus-49607.
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Gorgannezhad, Lena. "Advanced Technologies in Rapid and Multiplex Detection of Nucleic acid". Thesis, Griffith University, 2020. http://hdl.handle.net/10072/397045.
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Nucleic acids are key macromolecules of living organisms transferring genetic inheritance from one generation to the next. From how a living individual is created to how it interacts with external factors, all and all, can be found in the nucleic acid sequences inside every single cell of every organism. Therefore, the analysis of nucleic acids sequences is a critical capability for cancer and pathogen diagnoses, genotyping, and disease monitoring. To date, numerous methods have been used to detect both characterised and uncharacterised mutations and sequence variations. However, the detection of low amounts of mutant genes in the presence of high levels of wildtype sequences is still a challenge, and existing technologies has room for improvement. The classical approaches for nucleic acid detection and analyses mainly include DNA sequencing and the polymerase chain reaction (PCR). Although these methods with high analytical performance and reliability have facilitated the interrogation of the nucleic acids, some key obstacles such as the need of labelling, high costs for routine clinical use, slow turnaround time for giving results, the complexity of operation, and the inability to dually detect the genetic mutation in one step have limited their applications. To avoid drawbacks of the traditional approaches, a number of chip-based methods leverage electrochemical readouts or microfluidics to identify nucleic acids. However, there is still an unmet need for a less complex, rapid, low-cost, sensitive and accurate method to enable nucleic acid analysis even in resource-poor settings. The overall objective of this PhD thesis is to develop simple, inexpensive and accurate platforms for nucleic acid evaluation.
To achieve the aforementioned goal, the first attempt was to develop a lab-on-a-chip platform for cancer diagnosis by detection of circulating tumor nucleic acids (ctNAs) in plasma samples of cancer patients. ctNAs are fragmented DNA released from cancerous cells and tumours into the bloodstream of patients with cancer. Tumour-specific (epi-)genetic alterations in ctNAs are assumed to reflect tumour burden and could be of high value for cancer diagnosis, prognosis, and management. In the first part of the thesis, I developed a new electrochemical assay for the detection of FGFR2:FAM76A fusion gene in ctNAs extracted from ovarian cancer patients. The assay was based on the high electrocatalytic activity of a new class of superparamagnetic graphene-loaded iron oxide nanoparticles. Electrochemical detection demonstrated a limit of detection (LOD) as low as 1.0 fM, high specificity and excellent reproducibility.
In the second part of the thesis, I designed and developed a real-time and quantitative PCR system for microbial source tracking (MST) in water samples. MST is a DNA-based technology that enables water-quality managers to identify sources of faecal pollution in environmental waters. Most of the MST methodologies typically require specialized and costly equipment, elaborated and time-consuming operations as well as trained personnel. Here, a simple, low-cost, and sensitive platform was implemented on a microfluidic array chip. The array was successfully used for the real-time PCR-based multiplex detection of three human-associated MST markers (H8, Gen bac III, UidA). The PCR mixture was loaded into an array of channels in a single step utilising capillary filling without the need for liquid handling instruments. The array was then integrated with our custom-made thermal cycling and optical detection system. By employing the fabricated platform, the LOD of 71.8 DNA copies/μL was achieved for Gen bac III sequence. In summary, we introduced a sensitive, simple and economical real-time and quantitative PCR system for MST in water samples.
In a further study, I investigated how a nucleic acid amplification setup can be miniaturised. To reach this goal, I utilised liquid marbles as an ideal biochemical microreactors for targeted amplification of the NAs. Liquid marbles are formed by encapsulating microscale volume of liquid with a thin layer of hydrophobic particles. Miniaturization of the nucleic acids (NAs) amplification process inside a liquid droplet provides several advantages upon routine methods, such as reducing reagents consumption and contamination possibility, easy handling of liquids, eliminating the usage of disposable plastic consumables for carrying out biochemical reactions. However, one of the major concerns in liquid marble applications is the high rate of evaporation through the porous walls during the thermal cycling step. To eliminate the evaporation, I used core-shell beads synthesized from a composite liquid marble as a NAs amplification micro reactor comprising two non-miscible liquid droplets forming a spherical shape and a coating of hydrophobic powder. The shell liquid was then polymerised into a solid after exposure to blue light, converting the liquid marble into a core-shell bead. Fabricated core-shell beads were extended to explore their potential as a versatile bioreactor for phylogrouping of the E.coli strains. In general, this platform provided easy manipulation and storage of sample, elimination of the evaporation, and sample protection from possible external contamination. Moreover, this simple and effective method presented a sensitive and inexpensive way to track NAs.
In conclusion, this research endeavour presents a step forward towards the adaption of the selected group of tools and technologies, for the development of assays that can be applied as powerful alternatives to conventional tools used in molecular diagnostic. These technologies have the potential to revolutionise the NAs-based diagnostic approaches, by providing sensitive, rapid, accurate, and inexpensive platforms for point of care devices and in-field tests.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Environment and Sc
Science, Environment, Engineering and Technology
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Babair, Yasser Hassan Salih. "Multiplex PCR for diagnosis of herpes viruses in neurological infections". Thesis, University of Manchester, 2004. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.488092.
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Costa, Thadeu Estevam Moreira Maramaldo. "Detecção de transgênicos em alimentos utilizando a técnica Multiplex-PCR". reponame:Repositório Institucional da FIOCRUZ, 2008. https://www.arca.fiocruz.br/handle/icict/9250.
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Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde
Durante o período de 1996 a 2006, a proporção da área global de lavouras biotecnológicas plantadas por países em desenvolvimento aumentou consistentemente a cada ano. Nos últimos dez anos, mais de 40 países adotaram políticas de rotulagem para alimentos geneticamente modificados, mas as características das regulamentações e seu grau de execução variam enormemente. Os efeitos observados dessas políticas sobre a escolha dos consumidores, a informação para os consumidores, comércio dos alimentos, comércio internacional também variam significativamente. No Brasil, o Decreto n. 4.680/03 regulamenta o direito à informação, assegurado pela Lei n. 8.078/90, quanto aos alimentos e ingredientes alimentares destinados ao consumo humano ou animal que contenham ou sejam produzidos a partir de organismos geneticamente modificados (OGMs). O cumprimento da legislação que regulamenta a comercialização de alimentos e ingredientes contendo OGMs é totalmente dependente da sensibilidade e confiabilidade dos métodos de detecção e quantificação de OGMs. . Entre os métodos existentes, a reação em cadeia da polimerase (PCR) é o mais aceito, por ser sensível e confiável para detecção de material geneticamente modificado em análises de rotina. A padronização da técnica de PCR multiplex para a detecção de vários transgênicos em produtos alimentícios facilita o monitoramento destes, pois além de diminuir o tempo de diagnóstico, utiliza uma menor quantidade de reagentes, barateando o processo de detecção. No presente estudo, foi utilizada uma duplex PCR para a detecção de soja geneticamente modificada tolerante ao herbicida glifosato (Roundup Ready) e uma PCR multiplex para a detecção simultânea de três linhagens de milho transgênico (MON810, Bt11 e Bt176) em amostras de alimentos humano e animal. Foram utilizadas duas metodologias para extração do DNA das amostras: o método CTAB (já utilizado pelo INCQS) e o kit DNeasy Mini. [...]
From 1996 to 2006, the proportion of the global area of biotech crops growing in developing countries has increased consistently. In the last ten years, over 40 countries have adopted policies for the labeling of genetically modified food, but the regulations characteristics and their degree of implementation varies greatly. The effects of these policies on consumer choice and information, on food trade and international trade also vary significantly. In Brazil, the Decree No. 4.680/03 regulates the right to consumers information, provided by Law No. 8078/90, on food and food ingredients intended for human or animal consumption containing or produced from genetically modified organisms (GMOs). Compliance with regulation on the marketing of foods and ingredients containing GMOs is totally dependent on the sensitivity and reliability of methods of detection and quantification of GMOs. Among the existing methods, the polymerase chain reaction (PCR) is the most accepted, sensitive and reliable for the detection of genetically modified material in routine. The multiplex PCR is used on the screening of GMO in food because its able to detect simultaneously several transgenic, being a less time and reagents consuming method, making the process of detection cheaper. In the present study, duplex PCR was tested for the detection of genetically modified soybeans tolerant to the herbicide glyphosate (Roundup Ready) and a multiplex PCR was tested for the simultaneous detection of three lineages of transgenic maize (MON810, Bt11 and Bt176) in samples of human food and feed. Two methodologies were used for extraction of DNA from samples: the CTAB method (already used by INCQS) and DNeasy® Mini kit. The results were compared to the standard method used by INCQS. Out of the fourty samples, thirty samples were positive for the presence of Roundup Ready soybeans, fourteen were positive for at least one of the GM corn studied. Nine were positive for MON 810 maize, three were positive for Bt 11 maize, and five were positive for Bt 176 maize. The method proved to be robust and can be used on a routine for the detection of GMOs in food.
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Teixeira, Ana Izabel Passarella. "Desenvolvimento de uma PCR multiplex para identificação de clostrídios histotóxicos". Universidade Federal de Minas Gerais, 2012. http://hdl.handle.net/1843/BUOS-97BHV2.
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The polymerase chain reaction (PCR) has emerged in recent years as an important technique for identification of microorganisms to be extremely sensible and versatile. This work was the development
of a multiplex PCR for the detection and discrimination of histotoxic clostridia. The analytical sensitivity obtained was approximately 0.15 ng for C.chauvoeiand 0.015ng for other agents. No amplification occurred in the test of specificity for species Clostridium tetani(ATCC 9441), Staphylococcus aureus(ATCC 27707), Pseudomonas aeruginosa(ATCC 25319), Escherichia coli(ATCC 21986), Clostridium difficile(ATCC 9689), Proteus mirabilis(ATCC 12453) and Salmonella typhi(ATCC 9992V),
confirming it. This paper presents an efficient andspecific multiplex PCR for detection and identification of clostridia histotoxics.A reação em cadeia da polimerase (PCR) tem se destacado nos últimos anos como uma importante técnica para identificação de microrganismos por ser extremamente sensível e versátil. Porém, até então não existia uma PCR Multiplex que fosse capas de identificar os cinco agentes etiológicos causadores de mionecroses clostridiais: Clostridium septicum, C. chauvoei, C. novyi tipo A, C. perfringens tipo A e C. sordellii. Portanto, nesse trabalho foi desenvolvida uma PCR Multiplexpara a detecção e discriminação dos clostrídios histotóxicos. A sensibilidade analítica obtida foi de aproximadamente 0,15 ng para C.chauvoei e 0,015 ng para outros agentes. Não aconteceram amplificações no teste de especificidade com as espécies Clostridium tetani(ATCC 9441), Staphylococcus aureus (ATCC 27707), Pseudomonas aeruginosa (ATCC 25319), Escherichia coli (ATCC 21986), Clostridium difficile (ATCC 9689), Proteus mirabillis (ATCC 12453) e Salmonella typhi (ATCC 9992V) , confirmando-a. Este trabalho apresenta uma PCR Multiplex eficientee especifica para detecção e identificação de clostrídios histotóxicos.