999 resultados para Post-confluence
Resumo:
To investigate the role of peroxisome proliferator-activated receptors (PPARs) chi and beta in the differentiation of colon cancer cells, we differentiated HT-29 cells using sodium butyrate (NaB) and culturing post-confluence and assessed differentiation using the marker intestinal alkaline phosphatase. While PPAR chi levels only changed with culturing post confluence, PPAR beta levels increased independent of the method of differentiation. To explore further the differences induced by NaB. we assessed changes in both PPAR isoforms in MCF-7 breast cancer cells cultured in the presence of NaB over 48 h. Again a very different expression pattern was observed with PPAR-1 increasing after 4 h and remaining elevated, while PPAR beta increased transiently. Our studies suggest that the expression of PPARs is dependent upon both the method of differentiation and on time. Moreover, these studies show that changes in levels are not required for the differentiation of colon cancer cell lines, whereas changes in PPAR beta are more closely associated with differentiation. (c) 2005 Elsevier Inc. All rights reserved.
Resumo:
Calcium transporters play vital roles in the transport of calcium ions across cells of the mammary gland and the intestine. One such transporter is the plasma membrane Ca2+-ATPase (PMCA), of which there are 4 different genes (PMCA1-4). In these studies we investigated the hypothesis that the expression of PMCA is altered in HT-29 colon cancer cells during sodium butyrate and post-confluence mediated differentiation. We also investigated if PMCA expression is altered in breast cancer cell lines in an isofrom specific manner. Our results indicate isoform specific changes in PMCA mRNA and protein levels in HT-29 cells during differentiation, using real time RT-PCR and western blotting, respectively. We also observed pronounced alterations in the mRNA levels of the PMCA isoform linked to lactation (PMCA2) in a bank of breast cancer cell lines compared to normal cell lines. Changes in other isoforms were less pronounced. To further study the role of specific calcium transporters we have optimised conditions for the reverse transfection of MCF-7 breast cancer cells using NeoFX (Ambion). Using real time RT-PCR we have confirmed gene knockdown for specific isoforms and have studied the time course of knockdown over 96 hours. We see approximately 68 % inhibition at 24 hours increasing to 84 % 96 hours post-reverse transfection. Our studies suggest that the expression of specific calcium transporter isoforms can be significantly altered in cancer cell lines and that isoform specific inhibition of calcium transporters is possible using reverse transfection of siRNA
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Smooth muscle cell (SMC) phenotypic modulation from the mature ’contractile’ to a less differentiated ’synthetic’ phenotype involves not only altered expression but also a reorganisation of contractile and cytoskeletal proteins. Objective: To investigate the role of RhoA, a known regulator of the actin cytoskeleton, in SMC phenotypic regulation. Methods: Rho transcription (RT-PCR), expression (Western analysis) and activation (membrane translocation or Rho ’pull-down’ assay) was investigated in cultured rabbit aortic SMC during phenotypic modulation, and under the influence of known SM-regulatory proteins (thrombin, heparin and TGF- β). Rho’s effect on cell morphology was examined by transient transfection of ’synthetic’ state SMC with either constitutively active Rho (Val14RhoA) or its inhibitor, C3 transferase. Results: RhoA transcription was elevated in the first 3 days of primary culture, and protein expression peaked at 2 days post-confluence when SMC return to a more ’contractile’ state. However, RhoA showed augmented activation at three time-points in primary culture: the transition point when SMCs enter logarithmic growth and are highly motile, upon reaching quiescence, and when they return to a more ’contractile’ state. Thrombin, heparin and TGF-β activated RhoA in ’synthetic’ state SMCs. Transfection with Val14RhoA caused a dramatic decrease in SMC size and a reorganization of cytoskeletal proteins, reminiscent of the ’contractile’ phenotype. Specific inhibition of endogenous Rho by C3 transferase resulted in an almost complete loss of contractile proteins. Conclusion: These data indicate that Rho is an important determining factor of SMC functional state.
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The River Tawd is a tributary of the River Douglas catchment. The Tawd is approximately 8km in length from its source, to the south of Skelmersdale, to its confluence with the River Douglas at Snipe Hall farm. The aim of this study was to determine whether the water quality of the River Tawd had improved sufficiently to allow a mixed coarse fish population to establish itself on the river after the stocking of juvenile Roach (Rutilus rutilus), Dace (Leucisus leucisus) and Chub (Leucisus cephalus) in 1997. This report will consider the survival of the stocked fish. If the stocking was successful, other potential stocking sites will be recommended for future stocking with a view to developing the River Tawd fishery.
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La conception architecturale du logis, individuel ou collectif, est un défi majeur pour les architectes depuis l'avènement de la modernité. Au début du 21ième siècle, la multiplicité des conditions démographiques et celle de la mobilité des personnes sous-tendent la complexité de l'élaboration de prototypes ou de modèles d'habitation. Que peut-on apprendre des expériences menées dans ce domaine après la seconde guerre mondiale aux États-Unis ? Ce mémoire de maîtrise est consacré à l'étude d’un cas particulier celui du programme de création architecturale réalisée en Californie de 1945 à 1966, connu sous le nom de : «Case Study House Program». Ce programme, dirigé par John Entenza, éditeur de la revue Arts and Architecture, rassemblait de nombreux architectes dont les plus célèbres sont Charles et Ray Eames, Richard Neutra, Craig Ellwood et Pierre Koenig, tous auteurs de maisons modernes devenues oeuvres canoniques dans l’histoire de l’architecture. L'analyse détaillé de ce cas et de ses retombées devrait permettre de mieux cerner les aspects suivants: la portée critique du CSHP (case study house program) qui s'opposait aux modèles dominants du marché immobilier, modèles généralement inspirés de styles traditionnels; le potentiel et les limites d'une telle démarche face à la demande sociale; la dimension anticipatrice des propositions du CSHP pour la conception de logis mieux adaptés aux besoins du 21ième siècle, en particulier ceux qui découlent des changements démographiques et de la mobilité géographiques des personnes; la valeur d'exemple du CSHP pour mieux comprendre les fondements de la résistance du public aux innovations architecturales, autant du point de vue technique que du point de vue esthétique.
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Language is a unique aspect of human communication because it can be used to discuss itself in its own terms. For this reason, human societies potentially have superior capacities of co-ordination, reflexive self-correction, and innovation than other animal, physical or cybernetic systems. However, this analysis also reveals that language is interconnected with the economically and technologically mediated social sphere and hence is vulnerable to abstraction, objectification, reification, and therefore ideology – all of which are antithetical to its reflexive function, whilst paradoxically being a fundamental part of it. In particular, in capitalism, language is increasingly commodified within the social domains created and affected by ubiquitous communication technologies. The advent of the so-called ‘knowledge economy’ implicates exchangeable forms of thought (language) as the fundamental commodities of this emerging system. The historical point at which a ‘knowledge economy’ emerges, then, is the critical point at which thought itself becomes a commodified ‘thing’, and language becomes its “objective” means of exchange. However, the processes by which such commodification and objectification occurs obscures the unique social relations within which these language commodities are produced. The latest economic phase of capitalism – the knowledge economy – and the obfuscating trajectory which accompanies it, we argue, is destroying the reflexive capacity of language particularly through the process of commodification. This can be seen in that the language practices that have emerged in conjunction with digital technologies are increasingly non-reflexive and therefore less capable of self-critical, conscious change.
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five:fashion musings is an upcoming 2009 publicaton to celebrate the fashion discipline's five-year milestone at the Queensland Unviesity of Technology. it represents a body of work by fashion practitioners, aceademic and educators commissioned to explore their research in fashion theory, practice and pedogogy through five key themes.
Resumo:
The primary aims of scoliosis surgery are to halt the progression of the deformity, and to reduce its severity (cosmesis). Currently, deformity correction is measured in terms of posterior parameters (Cobb angles and rib hump), even though the cosmetic concern for most patients is anterior chest wall deformity. In this study, we propose a new measure for assessing anterior chest wall deformity and examine the correlation between rib hump and the new measure. 22 sets of CT scans were retrieved from the QUT/Mater Paediatric Spinal Research Database. The Image J software (NIH) was used to manipulate formatted CT scans into 3-dimensional anterior chest wall reconstructions. A ‘chest wall angle’ was then measured in relation to the first sacral vertebral body. The chest wall angle was found to be a reliable tool in the analysis of chest wall deformity. No correlation was found between the new measure and rib hump angle. Since rib hump has been shown to correlate with vertebral rotation on CT, this suggests that there maybe no correlation between anterior and posterior deformity measures. While most surgical procedures will adequately address the coronal imbalance & posterior rib hump elements of scoliosis, they do not reliably alter the anterior chest wall shape. This implies that anterior chest wall deformity is to a large degree an intrinsic deformity, not directly related to vertebral rotation.
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The picturesque aesthetic in the work of Sir John Soane, architect and collector, resonates in the major work of his very personal practice – the development of his house museum, now the Soane Museum in Lincoln’s Inn Fields in London. Soane was actively involved with the debates, practices and proponents of picturesque and classical practices in architecture and landscape and his lectures reveal these influences in the making of The Soane, which was built to contain and present diverse collections of classical and contemporary art and architecture alongside scavenged curiosities. The Soane Museum has been described as a picturesque landscape, where a pictorial style, together with a carefully defined itinerary, has resulted in the ‘apotheosis of the Picturesque interior’. Soane also experimented with making mock ruinscapes within gardens, which led him to construct faux architectures alluding to archaeological practices based upon the ruin and the fragment. These ideas framed the making of interior landscapes expressed through spatial juxtapositions of room and corridor furnished with the collected object that characterise The Soane Museum. This paper is a personal journey through the Museum which describes and then reviews aspects of Soane’s work in the context of contemporary theories on ‘new’ museology. It describes the underpinning picturesque practices that Soane employed to exceed the boundaries between interior and exterior landscapes and the collection. It then applies particular picturesque principles drawn from visiting The Soane to a speculative project for a house/landscape museum for the Oratunga historic property in outback South Australia, where the often, normalising effects of conservation practices are reviewed using minimal architectural intervention through a celebration of ruinous states.