The Proximity Ligation Assay (PLA) is a relatively new method which utilizes the strength of both immunoassays and DNA detection. PLA has the capacity of high multiplexing due to the high specificity achieved with both dual protein-binding and dual primer binding during detection with Real-Time PCR. We developed a multiplexed PLA protocol that can measure 28 biomarkers in human EDTA plasma. The method was tested on 46 individuals diagnosed with colorectal cancer and 48 age matched healthy controls. The results are very promising as we re-discover the most well-known biomarkers for colorectal cancer and also find some potential new markers (significance tested with students T-test with p<0.05). Further improvements of the protocol are needed to decrease the variation.

The work offered for this PhD by Published Works charts the history of cinema exhibition in Britain from the late 1950s to the present. At the start of this period, cinemagoing as a form of public entertainment entered a long period of decline that was only arrested with the development and growth of multiplex cinemas in the 1980s and 1990s. Despite these changes, the feature film itself remained a culturally and commercially valuable artefact, though increasingly this meant the Hollywood film. Whilst due consideration is afforded to the technological changes in cinemas and the cinema apparatus, my work places the development of cinemagoing in a broad social, economic, cultural and political context, and explains how these issues impact upon on-going developments. In the late 1950s, cinemagoing declined partly in response to changing leisure habits, demographic shifts, the growth of consumer culture, television, and the widespread adoption of new broadcast technologies like home video and satellite. The multiplex returned feature films to cinemas, but was a definitively American commercial form closely associated with new forms of leisure and out-of-town retailing. There are also parallels between the context for development of the multiplex in the USA – suburbanisation, shopping malls and reliance on the motorcar – and developments in Britain in the last 30 years. To this end there is a specific emphasis on the development of the multiplex cinema as part of a wider narrative about the re-positioning of cinemagoing as a collective, public form of visual entertainment, in the period from the mid-1980s, in the context of some dramatic changes in the transient nature of capitalism and urban planning. From the early 1990s onwards there was a growing anxiety about the impact of out-of-town developments on Britain’s urban centres, and a concomitant and renewed emphasis on the importance of the urban core rather than the edge. Thus, the key to understanding the evolution of cinema exhibition today is to pay particular attention to urban planning as inherently ideological, shifting and changing in line with broader political, economic and social considerations.

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.

Lygus Hahn (Hemiptera: Miridae) are serious pests of economically important field, fruit, vegetable, and greenhouse crops in Canada. The release of European Peristenus Förster (Hymenoptera: Braconidae) in the USA has resulted in significant suppression of this pest and has renewed interest in the release of European Peristenus spp. in Canada. Prior to the release of exotic Peristenus spp., ecological host range studies need to be conducted to define their habitat and host associations. <p>These associations can be difficult to study using conventional methods. Morphological similarity of related parasitoids prevents species-level identification by dissection. Host rearing is time-consuming and can result in high levels of host and parasitoid mortality. To facilitate identification of immature Peristenus spp. in their hosts, a multiplex PCR assay was developed. This assay provided a specific and sensitive tool to screen individual insects for three parasitoid species simultaneously. <p>To validate the utility of the multiplex PCR assay in ecological host range studies, parasitism and parasitoid species composition obtained using conventional and molecular techniques were compared. Molecular methods compared favorably with conventional methods; however, more complete species composition information was available with the multiplex assay. To improve the quality of risk assessment studies and extract the most accurate ecological host range data, molecular methods were used to evaluate host-parasitoid associations in mirid populations collected in two ecoregions. Several new host-parasitoid associations were recorded for <i>P. digoneutis</i> and <i>P. relictus</i>, but parasitism of non-target mirids was low. <p>Parasitism of the target host collected from different plant species was evaluated to help clarify Peristenus host-plant associations. Despite the investigation of three different host plant species, no difference was observed in the parasitism level or parasitoid species composition in <i>L. rugulipennis</i>. The post-release utility of the multiplex assay was investigated in Canada, where Lygus parasitoids may have dispersed following release in the USA. To confirm establishment, samples were analyzed using the multiplex PCR assay, and P. digoneutis was detected for the first time in southern Ontario.

Lygus Hahn (Hemiptera: Miridae) are serious pests of economically important field, fruit, vegetable, and greenhouse crops in Canada. The release of European Peristenus Förster (Hymenoptera: Braconidae) in the USA has resulted in significant suppression of this pest and has renewed interest in the release of European Peristenus spp. in Canada. Prior to the release of exotic Peristenus spp., ecological host range studies need to be conducted to define their habitat and host associations. <p>These associations can be difficult to study using conventional methods. Morphological similarity of related parasitoids prevents species-level identification by dissection. Host rearing is time-consuming and can result in high levels of host and parasitoid mortality. To facilitate identification of immature Peristenus spp. in their hosts, a multiplex PCR assay was developed. This assay provided a specific and sensitive tool to screen individual insects for three parasitoid species simultaneously. <p>To validate the utility of the multiplex PCR assay in ecological host range studies, parasitism and parasitoid species composition obtained using conventional and molecular techniques were compared. Molecular methods compared favorably with conventional methods; however, more complete species composition information was available with the multiplex assay. To improve the quality of risk assessment studies and extract the most accurate ecological host range data, molecular methods were used to evaluate host-parasitoid associations in mirid populations collected in two ecoregions. Several new host-parasitoid associations were recorded for <i>P. digoneutis</i> and <i>P. relictus</i>, but parasitism of non-target mirids was low. <p>Parasitism of the target host collected from different plant species was evaluated to help clarify Peristenus host-plant associations. Despite the investigation of three different host plant species, no difference was observed in the parasitism level or parasitoid species composition in <i>L. rugulipennis</i>. The post-release utility of the multiplex assay was investigated in Canada, where Lygus parasitoids may have dispersed following release in the USA. To confirm establishment, samples were analyzed using the multiplex PCR assay, and P. digoneutis was detected for the first time in southern Ontario.

The multiplex PCR experiment is to amplify multiple regions of a DNA sequence at the same time by using different primer pairs. Although, in recent years, there are lots of methods for PCR primer design, only a few of them focus on the multiplex PCR primer design. The multiplex PCR primer design is a tedious task since there are too many constraints to be satisfied. A new method for multiplex PCR primer design strategy using genetic algorithm is proposed. The proposed algorithm is able to find a set of suitable primer pairs more efficient and uses a MAP model to speed up the examination of the specificity constraint. The dry-dock experiment shows that the proposed algorithm finds several sets of primer pairs for multiplex PCR that not only obey the design properties, but also have specificity.

260 leaves : ill., graphs, tables ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Electrical Engineering, 1980

Stuttgart, FH, Diplomarb., 2000.

Made available in DSpace on 2014-06-11T19:30:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-12-13Bitstream added on 2014-06-13T20:21:07Z : No. of bitstreams: 1 braoios_a_dr_arafcf.pdf: 1217984 bytes, checksum: 5239ef0501af6f189fd2b422aad8dc7d (MD5) / Universidade Estadual Paulista (UNESP) / Staphylococcus aureus é um dos principais agentes de infecção humana, especialmente em indivíduos hospitalizados. Cepas MRSA (Methicillin Resistant Staphylococcus aureus) constituem um grave problema em hospitais de todo o mundo, aumentando a morbidade e mortalidade de indivíduos infectados. A vancomicina é uma das únicas opções terapêuticas. No entanto, o uso excessivo desse antibiótico pode selecionar cepas resistentes, agravando ainda mais o problema. Trabalhadores hospitalares podem carrear S. aureus nas narinas anteriores e pele e, assim, constituem um importante elo na epidemiologia das infecções nosocomiais. Nesse trabalho foram coletadas amostras das mãos e narinas de 100 trabalhadores do Hospital Universitário Dr. Domingos Leonardo Cerávolo da Universidade do Oeste Paulista - UNOESTE, em Presidente Prudente (SP). Desse total, 68% não eram portadores de S. aureus, 27% carreavam S. aureus sensível a meticilina, 4% carreavam MRSA e 1% carreava BORSA (Borderline Oxacillin Resistant Staphylococcus aureus). No mesmo período (julho a dezembro de 2002), 54,3% das infecções estafilocócicas em pacientes internados nessa Instituição tinham como agente MRSA. A técnica da PCR (Polymerase Chain Reaction) Multiplex para a detecção do gene femA (gene espécie-específico), mecA (resistência à meticilina) e ileS-2 (resistência à mupirocina) foi comparada com o método da difusão com discos e provas convencionais de identificação. Os resultados da PCR Multiplex apresentaram completa concordância com os resultados obtidos com os testes fenotípicos convencionais. Para verificar a relação genética entre as 30 cepas MRSA, 5 isoladas de trabalhadores e 25 de pacientes, foram realizadas a antibiotipagem e tipagem molecular através da técnica RAPD (Random Amplified Polymorphic DNA). Pela antibiotipagem as cepas MRSA foram... . / Staphylococcus aureus is considered as a major infeccious disease agent in human, especially in hospitalized people. Methicillin-resistant strains (MRSA) constitute a serious problem in hospitals around the world, increasing the morbity and mortality rate of infected people. Usually, vancomycin is the only therapeutic choice for the treatment of MRSA infections. However, excessive use of this antibiotic can select resistant strains, aggravating the problem. Health workers can carry S. aureus in the anterior nares and skin, constituting an important link for the epidemiology of nosocomial infections. In this study, samples of the hands and nares of one hundred workers from the University Hospital Dr. Domingos Leonardo Cerávolo , at Presidente Prudente, SP, Brazil, were assessed. Among the one hundred workers, 68% did not carry S. aureus, methicillin-susceptible S. aureus was found in 27/100 (27%) of those people; MRSA in 4/100 (4%) and BORSA (Borderline Resistant Staphylococcus aureus) was found in 1 (1%) health worker. In the same period (July to December of 2002), 54,3% of the staphylococcal infections in hospitalized patients were caused by MRSA. Multiplex PCR assay for the detection of femA (species-specific gene), mecA (methicillin resistance gene) and ileS-2 (mupirocin resistance gene) was compared to the disk diffusion susceptibility test and conventional identification test methods. The Multiplex PCR technic results were in complete agreement with the results obtained from conventional methods. Genetic relationship among the 30 MRSA strains, five isolated from workers and twenty five from patients, was established by antibiotyping and molecular typing by RAPD. On the basis of the antibiotyping, the 30 strains were grouped into four clusters (A to D), of a which 87% were grouped into antibiotype A. According to the profile of RAPD, eight clusters... (Complete abstract, click electronic address below).

Orientador : Aldario Chrestani Bordonalli / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Eletrica e de Computação / Made available in DSpace on 2018-08-02T00:51:33Z (GMT). No. of bitstreams: 1 Goncalves_MarcosSergio_M.pdf: 2370554 bytes, checksum: 642f9a0c1af60c1af182ef3f04482395 (MD5) Previous issue date: 2002 / Mestrado