966 resultados para hydrophobic interaction chromatography
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Fibre Bragg Grating (FBG) sensors have been installed along an existing line for the purposes of train detection and weight measurement. The results show fair accuracy and high resolution on the vertical force acted on track when the train wheels are rolling upon. While the sensors are already in place and data is available, further applications beyond train detection are explored. This study presents the analysis on the unique signatures from the data collected to characterise wheel-rail interaction for rail defect detection. Focus of this first stage of work is placed on the repeatability of signals from the same wheel-rail interactions while the rail is in healthy state. Discussions on the preliminary results and hence the feasibility of this condition monitoring application, as well as technical issues to be addressed in practice, are given.
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Presented as part of the Sampled Festival at Sadlers Wells UK in January 2009, PIPP #2 continues the exploration of the first installation (PIPP #1 Leeds) and asks audiences to connect with an interactive work presented in the foyer of a major dance festival. Literally choreographing their own dances on the walls of the venue, pedestrians re-connect with the architectural surrounds generating unique memories of self.
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The Arabidopsis thaliana NPR1 has been shown to be a key regulator of gene expression during the onset of a plant disease-resistance response known as systemic acquired resistance. The npr1 mutant plants fail to respond to systemic acquired resistance-inducing signals such as salicylic acid (SA), or express SA-induced pathogenesis-related (PR) genes. Using NPR1 as bait in a yeast two-hybrid screen, we identified a subclass of transcription factors in the basic leucine zipper protein family (AHBP-1b and TGA6) and showed that they interact specifically in yeast and in vitro with NPR1. Point mutations that abolish the NPR1 function in A. thaliana also impair the interactions between NPR1 and the transcription factors in the yeast two-hybrid assay. Furthermore, a gel mobility shift assay showed that the purified transcription factor protein, AHBP-1b, binds specifically to an SA-responsive promoter element of the A. thaliana PR-1 gene. These data suggest that NPR1 may regulate PR-1 gene expression by interacting with a subclass of basic leucine zipper protein transcription factors.
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As more and more information is available on the Web finding quality and reliable information is becoming harder. To help solve this problem, Web search models need to incorporate users’ cognitive styles. This paper reports the preliminary results from a user study exploring the relationships between Web users’ searching behavior and their cognitive style. The data was collected using a questionnaire, Web search logs and think-aloud strategy. The preliminary findings reveal a number of cognitive factors, such as information searching processes, results evaluations and cognitive style, having an influence on users’ Web searching behavior. Among these factors, the cognitive style of the user was observed to have a greater impact. Based on the key findings, a conceptual model of Web searching and cognitive styles is presented.
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With an increasing body of literature linking the human resource management and marketing fields, one area receiving increased academic attention is how an organisation’s corporate reputation can be managed to attract potential recruits and shape their employment expectations through their psychological contracts. This paper seeks to enhance current models which focus on the interrelationship of corporate reputation and psychological contract theory. It is argued that a number of factors need to be considered in order the build a firmer foundation for such a theory. Firstly, a common understanding of the psychological contract needs to be established such that the focus on either expectations or promises is clarified. Secondly, the included components of the psychological contract need to be considered in light of their empirical founding and their relationship with one another. Thirdly, the interrelationship of corporate reputation, employer branding, identity and image needs to be explicated within the context of how they both influence and interrelate with the psychological contract. The final consideration surrounds the opportunity for potential employees to be considered within the corporate reputation literature as a significant stakeholder group.
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Prostate cancer is the second most common cause of cancer-related deaths in Western males. Current diagnostic, prognostic and treatment approaches are not ideal and advanced metastatic prostate cancer is incurable. There is an urgent need for improved adjunctive therapies and markers for this disease. GPCRs are likely to play a significant role in the initiation and progression of prostate cancer. Over the last decade, it has emerged that G protein coupled receptors (GPCRs) are likely to function as homodimers and heterodimers. Heterodimerisation between GPCRs can result in the formation of novel pharmacological receptors with altered functional outcomes, and a number of GPCR heterodimers have been implicated in the pathogenesis of human disease. Importantly, novel GPCR heterodimers represent potential new targets for the development of more specific therapeutic drugs. Ghrelin is a 28 amino acid peptide hormone which has a unique n-octanoic acid post-translational modification. Ghrelin has a number of important physiological roles, including roles in appetite regulation and the stimulation of growth hormone release. The ghrelin receptor is the growth hormone secretagogue receptor type 1a, GHS-R1a, a seven transmembrane domain GPCR, and GHS-R1b is a C-terminally truncated isoform of the ghrelin receptor, consisting of five transmembrane domains. Growing evidence suggests that ghrelin and the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, may have a role in the progression of a number of cancers, including prostate cancer. Previous studies by our research group have shown that the truncated ghrelin receptor isoform, GHS-R1b, is not expressed in normal prostate, however, it is expressed in prostate cancer. The altered expression of this truncated isoform may reflect a difference between a normal and cancerous state. A number of mutant GPCRs have been shown to regulate the function of their corresponding wild-type receptors. Therefore, we investigated the potential role of interactions between GHS-R1a and GHS-R1b, which are co-expressed in prostate cancer and aimed to investigate the function of this potentially new pharmacological receptor. In 2005, obestatin, a 23 amino acid C-terminally amidated peptide derived from preproghrelin was identified and was described as opposing the stimulating effects of ghrelin on appetite and food intake. GPR39, an orphan GPCR which is closely related to the ghrelin receptor, was identified as the endogenous receptor for obestatin. Recently, however, the ability of obestatin to oppose the effects of ghrelin on appetite and food intake has been questioned, and furthermore, it appears that GPR39 may in fact not be the obestatin receptor. The role of GPR39 in the prostate is of interest, however, as it is a zinc receptor. Zinc has a unique role in the biology of the prostate, where it is normally accumulated at high levels, and zinc accumulation is altered in the development of prostate malignancy. Ghrelin and zinc have important roles in prostate cancer and dimerisation of their receptors may have novel roles in malignant prostate cells. The aim of the current study, therefore, was to demonstrate the formation of GHS-R1a/GHS-R1b and GHS-R1a/GPR39 heterodimers and to investigate potential functions of these heterodimers in prostate cancer cell lines. To demonstrate dimerisation we first employed a classical co-immunoprecipitation technique. Using cells co-overexpressing FLAG- and Myc- tagged GHS-R1a, GHS-R1b and GPR39, we were able to co-immunoprecipitate these receptors. Significantly, however, the receptors formed high molecular weight aggregates. A number of questions have been raised over the propensity of GPCRs to aggregate during co-immunoprecipitation as a result of their hydrophobic nature and this may be misinterpreted as receptor dimerisation. As we observed significant receptor aggregation in this study, we used additional methods to confirm the specificity of these putative GPCR interactions. We used two different resonance energy transfer (RET) methods; bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), to investigate interactions between the ghrelin receptor isoforms and GPR39. RET is the transfer of energy from a donor fluorophore to an acceptor fluorophore when they are in close proximity, and RET methods are, therefore, applicable to the observation of specific protein-protein interactions. Extensive studies using the second generation bioluminescence resonance energy transfer (BRET2) technology were performed, however, a number of technical limitations were observed. The substrate used during BRET2 studies, coelenterazine 400a, has a low quantum yield and rapid signal decay. This study highlighted the requirement for the expression of donor and acceptor tagged receptors at high levels so that a BRET ratio can be determined. After performing a number of BRET2 experimental controls, our BRET2 data did not fit the predicted results for a specific interaction between these receptors. The interactions that we observed may in fact represent ‘bystander BRET’ resulting from high levels of expression, forcing the donor and acceptor into close proximity. Our FRET studies employed two different FRET techniques, acceptor photobleaching FRET and sensitised emission FRET measured by flow cytometry. We were unable to observe any significant FRET, or FRET values that were likely to result from specific receptor dimerisation between GHS-R1a, GHS-R1b and GPR39. While we were unable to conclusively demonstrate direct dimerisation between GHS-R1a, GHS-R1b and GPR39 using several methods, our findings do not exclude the possibility that these receptors interact. We aimed to investigate if co-expression of combinations of these receptors had functional effects in prostate cancers cells. It has previously been demonstrated that ghrelin stimulates cell proliferation in prostate cancer cell lines, through ERK1/2 activation, and GPR39 can stimulate ERK1/2 signalling in response to zinc treatments. Additionally, both GHS-R1a and GPR39 display a high level of constitutive signalling and these constitutively active receptors can attenuate apoptosis when overexpressed individually in some cell types. We, therefore, investigated ERK1/2 and AKT signalling and cell survival in prostate cancer the potential modulation of these functions by dimerisation between GHS-R1a, GHS-R1b and GPR39. Expression of these receptors in the PC-3 prostate cancer cell line, either alone or in combination, did not alter constitutive ERK1/2 or AKT signalling, basal apoptosis or tunicamycin-stimulated apoptosis, compared to controls. In summary, the potential interactions between the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, and the related zinc receptor, GPR39, and the potential for functional outcomes in prostate cancer were investigated using a number of independent methods. We did not definitively demonstrate the formation of these dimers using a number of state of the art methods to directly demonstrate receptor-receptor interactions. We investigated a number of potential functions of GPR39 and GHS-R1a in the prostate and did not observe altered function in response to co-expression of these receptors. The technical questions raised by this study highlight the requirement for the application of extensive controls when using current methods for the demonstration of GPCR dimerisation. Similar findings in this field reflect the current controversy surrounding the investigation of GPCR dimerisation. Although GHS-R1a/GHS-R1b or GHS-R1a/GPR39 heterodimerisation was not clearly demonstrated, this study provides a basis for future investigations of these receptors in prostate cancer. Additionally, the results presented in this study and growing evidence in the literature highlight the requirement for an extensive understanding of the experimental method and the performance of a range of controls to avoid the spurious interpretation of data gained from artificial expression systems. The future development of more robust techniques for investigating GPCR dimerisation is clearly required and will enable us to elucidate whether GHS-R1a, GHS-R1b and GPR39 form physiologically relevant dimers.
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Agile ridesharing aims to utilise the capability of social networks and mobile phones to facilitate people to share vehicles and travel in real time. However the application of social networking technologies in local communities to address issues of personal transport faces significant design challenges. In this paper we describe an iterative design-based approach to exploring this problem and discuss findings from the use of an early prototype. The findings focus upon interaction, privacy and profiling. Our early results suggest that explicitly entering information such as ride data and personal profile data into formal fields for explicit computation of matches, as is done in many systems, may not be the best strategy. It might be preferable to support informal communication and negotiation with text search techniques.
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This paper introduces Sapporo World Window, a screen-based application that is currently under development for the new underway passage at the centre of Sapporo City. There are ten large public screens installed in the space, displaying user-generated videos about various aspects of the city and a real-time map that visualises users’ interaction with the city. The application aims to engage the general public by functioning as a unique ‘point of connection’ for socio-cultural and technological interactions, making the space a lively social place where people can have meaningful experiences of interacting with people and places of Sapporo through mobile phones (keitai) and the public screens in the space. This paper first outlines the contextual background and key concept for the application’s design. Then the paper discusses the user interaction processes, technical specifications, and interface design, followed by the conclusions and outlook.
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Corepressors play a crucial role in negative gene regulation and are defective in several diseases. BCoR is a corepressor for the BCL6 repressor protein. Here we describe and functionally characterize BCoR-L1, a homolog of BCoR. When tethered to a heterologous promoter, BCoR-L1 is capable of strong repression. Like other corepressors, BCoR-L1 associates with histone deacetylase (HDAC) activity. Specifically, BCoR-L1 coprecipitates with the Class II HDACs, HDAC4, HDAC5, and HDAC7, suggesting that they are involved in its role as a transcriptional repressor. BCoR-L1 also interacts with the CtBP corepressor through a CtBP-interacting motif in its amino terminus. Abrogation of the CtBP binding site within BCoR-L1 partially relieves BCoR-L1-mediated transcriptional repression. Furthermore, BCoR-L1 is located on the E-cadherin promoter, a known CtBP-regulated promoter, and represses the E-cadherin promoter activity in a reporter assay. The inhibition of BCoR-L1 expression by RNA-mediated interference results in derepression of E-cadherin in cells that do not normally express E-cadherin, indicating that BCoR-L1 contributes to the repression of an authentic endogenous CtBP target.
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Archaeal transcription utilizes a complex multisubunit RNA polymerase and the basal transcription factors TBP and TF(II)B, closely resembling its eukaryal counterpart. We have uncovered a tight physical and functional interaction between RNA polymerase and the single-stranded DNA-binding protein SSB in Sulfolobus solfataricus. SSB stimulates transcription from promoters in vitro under TBP-limiting conditions and supports transcription in the absence of TBP. SSB also rescues transcription from repression by reconstituted chromatin. We demonstrate the potential for promoter melting by SSB, suggesting a plausible basis for the stimulation of transcription. This stimulation requires both the single-stranded DNA-binding domain and the acidic C-terminal tail of the SSB. The tail forms a stable interaction with RNA polymerase. These data reveal an unexpected role for single-stranded DNA-binding proteins in transcription in archaea.
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Many luxury heritage brands operate on the misconception that heritage is interchangeable with history rather than representative of the emotional response they originally developed in their customer. This idea of heritage as static history inhibits innovation, prevents dynamic renewal and impedes their ability to redefine, strengthen and position their brand in current and emerging marketplaces. This paper examines a number of heritage luxury brands that have successfully identified the original emotional responses they developed in their customers and, through innovative approaches in design, marketing, branding and distribution evoke these responses in contemporary consumers. Using heritage and innovation hand-in-hand, these brands have continued to grow and develop a vision of heritage that incorporates both historical and contemporary ideas to meet emerging customer needs. While what constitutes a ‘luxury’ item is constantly challenged in this era of accessible luxury products, up-scaling and aspirational spending, this paper sees consumers’ emotional needs as the key element in defining the concept of luxury. These emotional qualities consistently remain relevant due to their ability to enhance a positive sense of identity for the brand user. Luxury is about the ‘experience’ not just the product providing the consumer with a sense of enhanced status or identity through invoked feelings of exclusivity, authenticity, quality, uniqueness and culture. This paper will analyse luxury heritage brands that have successfully combined these emotional values with those of their ‘heritage’ to create an aura of authenticity and nostalgia that appeals to contemporary consumers. Like luxury, the line where clothing becomes fashion is blurred in the contemporary fashion industry; however, consumer emotion again plays an important role. For example, clothing becomes ‘fashion’ for consumers when it affects their self perception rather than fulfilling basic functions of shelter and protection. Successful luxury heritage brands can enhance consumers’ sense of self by involving them in the ‘experience’ and ‘personality’ of the brand so they see it as a reflection of their own exclusiveness, authentic uniqueness, belonging and cultural value. Innovation is a valuable tool for heritage luxury brands to successfully generate these desired emotional responses and meet the evolving needs of contemporary consumers. While traditionally fashion has been a monologue from brand to consumer, new technology has given consumers a voice to engage brands in a conversation to express their evolving needs, ideas and feedback. As a result, in this consumer-empowered era of information sharing, this paper defines innovation as the ability of heritage luxury brands to develop new design and branding strategies in response to this consumer feedback while retaining the emotional core values of their heritage. This paper analyses how luxury heritage brands can effectively position themselves in the contemporary marketplace by separating heritage from history to incorporate innovative strategies that will appeal to consumer needs of today and tomorrow.
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Interaction Design is a fast developing branch of Industrial Design. The availability of cheap microprocessors and sensor electronics allow interactions between people and products that were until recently impossible. This has added additional layers of complexity to the design process. Novice designers find it difficult to effectively juggle these complexities and typically tend to focus on one aspect at a time. They also tend to take a linear, step-by-step approach to the design process in contrast to expert designers who pursue “parallel lines of thought” whilst simultaneously co-evolving both problem and solution. (Lawson, 1993) This paper explores an approach that encourages designers (in this case novice designers) to take a parallel rather than linear approach to the design process. It also addresses the problem of social loafing that tends to occur in team activities.
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The interaction of 10-hydroxycamptothecine (HCPT) with DNA under pseudo-physiological conditions (Tris-HCl buffer of pH 7.4), using ethidium bromide (EB) dye as a probe, was investigated with the use of spectrofluorimetry, UV-vis spectrometry and viscosity measurement. The binding constant and binding number for HCPT with DNA were evaluated as (7.1 ± 0.5) × 104 M-1 and 1.1, respectively, by multivariate curve resolution-alternating least squares (MCR-ALS). Moreover, parallel factor analysis (PARAFAC) was applied to resolve the three-way fluorescence data obtained from the interaction system, and the concentration information for the three components of the system at equilibrium was simultaneously obtained. It was found that there was a cooperative interaction between the HCPT-DNA complex and EB, which produced a ternary complex of HCPT-DNA-EB. © 2011 Elsevier B.V.
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The broad research questions of the book are: How can successful, interdisciplinary collaboration contribute to research innovation through Practice-led research? What contributes to the design, production and curation of successful new media art? What are the implications of exhibiting it across dual sites for artists, curators and participant audiences? Is it possible to create an 'intimate transaction' between people who are separated by vast distances but joined by interfaces and distributed networks? Centred on a new media work of the same name by the Transmute Collective (led by Keith Armstrong), this book provides insights from multidisciplinary perspectives. Visual, sound and performance artists, furniture designers, spatial architects, technology systems designers, and curators who collaborated in the production of Intimate Transactions discuss their design philosophies, working processes and resolution of this major new media work. Analytical and philosophical essays by international writers complement these writings on production. They consider how new media art, like Intimate Transactions, challenges traditional understandings of art, curatorial installation and exhibition experience because of the need to take into account interaction, the reconfiguration of space, co-presence, performativity and inter-site collaboration.