Mapping Protein-DNA Interactions Using ChIP-exo and Illumina-Based Sequencing

Chromatin immunoprecipitation (ChIP) provides a means of enriching DNA associated with transcription factors, histone modifications, and indeed any other proteins for which suitably characterized antibodies are available. Over the years, sequence detection has progressed from quantitative real-time PCR and Southern blotting to microarrays (ChIP-chip) and now high-throughput sequencing (ChIP-seq). This progression has vastly increased the sequence coverage and data volumes generated. This in turn has enabled informaticians to predict the identity of multi-protein complexes on DNA based on the overrepresentation of sequence motifs in DNA enriched by ChIP with a single antibody against a single protein. In the course of the development of high-throughput sequencing, little has changed in the ChIP methodology until recently. In the last three years, a number of modifications have been made to the ChIP protocol with the goal of enhancing the sensitivity of the method and further reducing the levels of nonspecific background sequences in ChIPped samples. In this chapter, we provide a brief commentary on these methodological changes and describe a detailed ChIP-exo method able to generate narrower peaks and greater peak coverage from ChIPped material.

The Big Crunch : A Mapping of the Swedish Party System from 1991 to 2010

Previous research claims that there has been a narrowing of distance between the Swedish political parties. Typically, such research into political distance has primarily focused on studying voters rather than the political parties themselves. In this article, the author conducts a longitudinal analysis of Comparative Manifesto Project data to determine if, and to what extent, the political parties have converged ideologically on a Left-Right continuum in the period 1991-2010. After first unraveling the concept of political distance, the author moves on to explain why the ideological dispersion of political parties is an important and consequential characteristic within party systems. Furthermore, the author argues that the Left-Right ideological scale continues to be a highly useful model with which to conceptualize and study this characteristic. The author then discusses the methodological approach and explains why quantitative manifesto data, often overlooked in favor of voter interview data, is deemed a valid and reliable material for measuring the ideological positions of political parties. The findings are that there indeed have been over all tendencies of ideological convergence between the blocs and that, in terms of how political parties are dispersed on a Left- Right ideological continuum, by 2010, the Swedish party system (the Sweden Democrats excluded) had become much less polarized than it had been in 1991.

The Accuracy and Reliability of Traditional Surface Flow Type Mapping: Is It Time for A New Method of Characterising Physical River Habitat?

Surface flow types (SFT) are advocated as ecologically relevant hydraulic units, often mapped visually from the bankside to characterise rapidly the physical habitat of rivers. SFT mapping is simple, non-invasive and cost-efficient. However, it is also qualitative, subjective and plagued by difficulties in recording accurately the spatial extent of SFT units. Quantitative validation of the underlying physical habitat parameters is often lacking, and does not consistently differentiate between SFTs. Here, we investigate explicitly the accuracy, reliability and statistical separability of traditionally mapped SFTs as indicators of physical habitat, using independent, hydraulic and topographic data collected during three surveys of a c. 50m reach of the River Arrow, Warwickshire, England. We also explore the potential of a novel remote sensing approach, comprising a small unmanned aerial system (sUAS) and Structure-from-Motion photogrammetry (SfM), as an alternative method of physical habitat characterisation. Our key findings indicate that SFT mapping accuracy is highly variable, with overall mapping accuracy not exceeding 74%. Results from analysis of similarity (ANOSIM) tests found that strong differences did not exist between all SFT pairs. This leads us to question the suitability of SFTs for characterising physical habitat for river science and management applications. In contrast, the sUAS-SfM approach provided high resolution, spatially continuous, spatially explicit, quantitative measurements of water depth and point cloud roughness at the microscale (spatial scales ≤1m). Such data are acquired rapidly, inexpensively, and provide new opportunities for examining the heterogeneity of physical habitat over a range of spatial and temporal scales. Whilst continued refinement of the sUAS-SfM approach is required, we propose that this method offers an opportunity to move away from broad, mesoscale classifications of physical habitat (spatial scales 10-100m), and towards continuous, quantitative measurements of the continuum of hydraulic and geomorphic conditions which actually exists at the microscale.

Enhancing business English terminology through concept mapping

Given the need of a growing internationalization of business, to have a good command of English is, most of the times important for the development of technical (specific) competences. It is, thus, critical that professionals use accurate terminology to set grounds for a well-succeeded communication. Furthermore, business communication is increasingly moving to ICT-mediated sets and professionals have to be able to promptly adjust to these needs, resorting to trustworthy online information sources, but also using technologies that better serve their business purposes. In this scenario, the main objective of this study is to find evidence as to the utility of concept mapping as a teaching and learning strategy for the appropriation of business English terminology, enabling students to use English more efficiently as language of communication in business context. This study was based on a case study methodology, mainly of exploratory nature. Participants were students (n= 30) enrolled in the subject English Applied to Management II at Águeda School of Technology and Management – University of Aveiro (2013/14 edition). They were asked to create and peer review two concept maps (cmaps), one individually and another in pairs. The data gathered were treated and analysed resorting qualitative (content analysis) and to quantitative (descriptive statistical analysis) techniques. Results of the data analysis unveil that the use of a collaborative concept mapping tool promotes the development of linguistic competences as to the use of business terminology, but also of communication and collaboration competences. Besides, it was also a very important motivation element in the students’ engagement with the subject content.

Transcriptome Analysis of Capsicum Chlorosis Virus-Induced Hypersensitive Resistance Response in Bell Capsicum

Background Capsicum chlorosis virus (CaCV) is an emerging pathogen of capsicum, tomato and peanut crops in Australia and South-East Asia. Commercial capsicum cultivars with CaCV resistance are not yet available, but CaCV resistance identified in Capsicum chinense is being introgressed into commercial Bell capsicum. However, our knowledge of the molecular mechanisms leading to the resistance response to CaCV infection is limited. Therefore, transcriptome and expression profiling data provide an important resource to better understand CaCV resistance mechanisms. Methodology/Principal Findings We assembled capsicum transcriptomes and analysed gene expression using Illumina HiSeq platform combined with a tag-based digital gene expression system. Total RNA extracted from CaCV/mock inoculated CaCV resistant (R) and susceptible (S) capsicum at the time point when R line showed a strong hypersensitive response to CaCV infection was used in transcriptome assembly. Gene expression profiles of R and S capsicum in CaCV- and buffer-inoculated conditions were compared. None of the genes were differentially expressed (DE) between R and S cultivars when mock-inoculated, while 2484 genes were DE when inoculated with CaCV. Functional classification revealed that the most highly up-regulated DE genes in R capsicum included pathogenesis-related genes, cell death-associated genes, genes associated with hormone-mediated signalling pathways and genes encoding enzymes involved in synthesis of defense-related secondary metabolites. We selected 15 genes to confirm DE expression levels by real-time quantitative PCR. Conclusion/Significance DE transcript profiling data provided comprehensive gene expression information to gain an understanding of the underlying CaCV resistance mechanisms. Further, we identified candidate CaCV resistance genes in the CaCV-resistant C. annuum x C. chinense breeding line. This knowledge will be useful in future for fine mapping of the CaCV resistance locus and potential genetic engineering of resistance into CaCV-susceptible crops.

Mapping the Liminal Experience of Eastern Europe in Selected American Works, 1965-2002

The purpose of this dissertation is to study literary representations of Eastern Europe in the works of celebrated and less-known American authors, who visited and narrated the region between the mid-1960s and early 2000s. The main critical body focuses on Eastern Europe before 1989 and encompasses three major voices of American literature: John Updike, Joyce Carol Oates, and Philip Roth. However, in the last chapter I also explore American literary perceptions of the area following the collapse of communism. Importantly, the term “Eastern Europe” as used in this dissertation is charged with significance. I approach it not only as a space on the map or the geopolitical construct which emerged in the aftermath of the Second World War, but rather as a conceptual category and a repository of meanings built out of fact and fantasy: specific historical, political and cultural realities interlaced with subjective worldviews, preconceptions, and mental images. The critical framework of this dissertation is twofold. I reach for the concept of liminality to elucidate the indeterminacy and malleability which lies at the heart of the object of study—the idea, image, and experience of Eastern Europe. Bearing in mind the nature of the works under analysis, all of which were inspired by actual visits behind the Iron Curtain, I propose to interpret these transatlantic literary journeys in terms of generative experience, where Eastern Europe is mapped as a liminal space of possibility; a contact zone between cultures and, potentially, the locus of self-discovery and individual transformation. If liminality is the metaphor or a lens that I employ in order to account for the nature of the analyzed works and the complex terrain they map, imagology, whose purpose is to study the processes of constructing selfhood and otherness in literature, provides me with the method and the critical vocabulary for analyzing selected literary representations. The dissertation is divided into six chapters, the last of which serves as coda to the previous discussion. The first two chapters constitute the critical foundation of this work. Then, in chapters 3, 4, and 5 I study American images of Eastern Europe in the works written by John Updike, Joyce Carol Oates, and Philip Roth, respectively. The last, sixth chapter of this dissertation is divided into two parts. In the first one, I discuss new critical perspectives and avenues of research in the study of Eastern Europe following the collapse of communism. Then, I carry out a joint analysis of four works written after 1989 by Eva Hoffman, Arthur Phillips, John Beckman, and Gary Shteyngart. The dissertation ends with conclusions in which I summarize my findings and reflections, and suggest implications for future research. As this dissertation seeks to demonstrate, Eastern Europe portrayed in the analyzed works oscillates between contradictory representations which are contingent upon a number of factors, most importantly who maps it and in what context. Even though each experience of Eastern Europe is distinct and fueled by the profiles, identities, and interests of the characters and their creators, I have found out that certain patterns of othering are present in all the works. Thus, my research seems to suggest that there is something of a recurrent literary image of Eastern Europe, which goes beyond the context of the Cold War. Accordingly, while this dissertation hopes to be a valid contribution to the study of literary and cultural mappings of Eastern Europe, it also generates new questions regarding the current, post-communist representation of the area and its relationship to the national tropes explored in my work.

The burden of neck pain: its meaning for persons with neck pain and healthcare providers, explored by concept mapping

PURPOSE: To empirically define the concept of burden of neck pain. The lack of a clear understanding of this construct from the perspective of persons with neck pain and care providers hampers adequate measurement of this burden. An additional aim was to compare the conceptual model obtained with the frequently used Neck Disability Index (NDI). METHODS: Concept mapping, combining qualitative (nominal group technique and group consensus) and quantitative research methods (cluster analysis and multidimensional scaling), was applied to groups of persons with neck pain (n = 3) and professionals treating persons with neck pain (n = 2). Group members generated statements, which were organized into concept maps. Group members achieved consensus about the number and description of domains and the researchers then generated an overall mind map covering the full breadth of the burden of neck pain. RESULTS: Concept mapping revealed 12 domains of burden of neck pain: impaired mobility neck, neck pain, fatigue/concentration, physical complaints, psychological aspects/consequences, activities of daily living, social participation, financial consequences, difficult to treat/difficult to diagnose, difference of opinion with care providers, incomprehension by social environment, and how person with neck pain deal with complaints. All ten items of the NDI could be linked to the mind map, but the NDI measures only part of the burden of neck pain. CONCLUSION: This study revealed the relevant domains for the burden of neck pain from the viewpoints of persons with neck pain and their care providers. These results can guide the identification of existing measurements instruments for each domain or the development of new ones to measure the burden of neck pain.

Optimal control of non-linear systems through hybrid cell-mapping/artificial-neural-networks techniques

In this paper, a real-time optimal control technique for non-linear plants is proposed. The control system makes use of the cell-mapping (CM) techniques, widely used for the global analysis of highly non-linear systems. The CM framework is employed for designing approximate optimal controllers via a control variable discretization. Furthermore, CM-based designs can be improved by the use of supervised feedforward artificial neural networks (ANNs), which have proved to be universal and efficient tools for function approximation, providing also very fast responses. The quantitative nature of the approximate CM solutions fits very well with ANNs characteristics. Here, we propose several control architectures which combine, in a different manner, supervised neural networks and CM control algorithms. On the one hand, different CM control laws computed for various target objectives can be employed for training a neural network, explicitly including the target information in the input vectors. This way, tracking problems, in addition to regulation ones, can be addressed in a fast and unified manner, obtaining smooth, averaged and global feedback control laws. On the other hand, adjoining CM and ANNs are also combined into a hybrid architecture to address problems where accuracy and real-time response are critical. Finally, some optimal control problems are solved with the proposed CM, neural and hybrid techniques, illustrating their good performance.

Zeitgeist or chameleon? A quantitative analysis of CSR definitions.

Despite its increasing relevance, corporate social responsibility (CSR) remains hobbled by problems, variously charged as being chameleon, vacuous or an utterly meaningless concept. One reason is the absence of an agreed upon normative basis underpinning CSR. This is in large part due to the concept lacking a universally accepted definition. This paper explores how the concept of CSR has evolved over time drawing from 110 definitions of the construct. Using co-word analysis of definitions from 1953 to 2014, the study maps how the structure of the definitions has evolved during the field's historical development. The research uncovers the key terms underpinning the phenomenon, the centrality of these terms as well as mapping their interrelationships and evolution. The findings suggest that, despite the profusion and definitional heterogeneity over the six decades of the development of the field, there are six recurrent, enduring dimensions that underpin the CSR concept. These dimensions are economic, social, ethical, stakeholders, sustainability and voluntary. This paper makes several contributions to the academic literature. The systematic, quantitative analysis of definitions brings an objectivity that previous qualitative bibliometric analyses of CSR have lacked. The time period selected is substantially longer than previous analyses and captures the complete historical evolution of the concept. Moreover, the analysis provides the basis for the development of a new, comprehensive, yet concise, definition of CSR that captures all six of the recurring dimensions underpinning the concept.

The Expanded Human Kallikrein (KLK) gene family : genomic organisation, tissue specific expression and potential functions

The tissue kallikreins are serine proteases encoded by highly conserved multigene families. The rodent kallikrein (KLK) families are particularly large, consisting of 13 26 genes clustered in one chromosomal locus. It has been recently recognised that the human KLK gene family is of a similar size (15 genes) with the identification of another 12 related genes (KLK4-KLK15) within and adjacent to the original human KLK locus (KLK1-3) on chromosome 19q13.4. The structural organisation and size of these new genes is similar to that of other KLK genes except for additional exons encoding 5 or 3 untranslated regions. Moreover, many of these genes have multiple mRNA transcripts, a trait not observed with rodent genes. Unlike all other kallikreins, the KLK4-KLK15 encoded proteases are less related (25–44%) and do not contain a conventional kallikrein loop. Clusters of genes exhibit high prostatic (KLK2-4, KLK15) or pancreatic (KLK6-13) expression, suggesting evolutionary conservation of elements conferring tissue specificity. These genes are also expressed, to varying degrees, in a wider range of tissues suggesting a functional involvement of these newer human kallikrein proteases in a diverse range of physiological processes.