PDGF regulates the actin cytoskeleton through hnRNP-K-mediated activation of the ubiquitin E-3-ligase MIR

PDGF is a potent chemotactic mitogen and a strong inductor of fibroblast motility. In Swiss 3T3 fibroblasts, exposure to PDGF but not EGF or IGF-1 causes a rapid loss of actin stress fibers (SFs) and focal adhesions (FAs), which is followed by the development of retractile dendritic protrusions and induction of motility. The PDGF-specific actin reorganization was blocked by inhibition of Src-kinase and the 26S proteasome. PDGF induced Src-dependent association between the multifunctional transcription/translation regulator hnRNP-K and the mRNA-encoding myosin regulatory light-chain (MRLC)-interacting protein (MIR), a E3-ubiquitin ligase that is MRLC specific. This in turn rapidly increased MIR expression, and led to ubiquitination and proteasome-mediated degradation of MRLC. Downregulation of MIR by RNA muting prevented the reorganization of actin structures and severely reduced the migratory and wound-healing potential of PDGF-treated cells. The results show that activation of MIR and the resulting removal of diphosphorylated MRLC are essential for PDGF to instigate and maintain control over the actin-myosin-based contractile system in Swiss 3T3 fibroblasts. The PDGF induced protein destabilization through the regulation of hnRNP-K controlled ubiquitin-ligase translation identifies a novel pathway by which external stimuli can regulate phenotypic development through rapid, organelle-specific changes in the activity and stability of cytoskeletal regulators.

Placing defects at the heart of high quality new homes: a learning perspective

The UK new-build housing sector is facing dual pressures to expand supply, whilst delivering against tougher planning and Building Regulation requirements; predominantly in the areas of sustainability. The sector is currently responding by significantly scaling up production and incorporating new technical solutions into new homes. This trajectory of up-scaling and technical innovation has been of research interest; but this research has primarily focus on the ‘upstream’ implications for house builders’ business models and standardised design templates. There has been little attention, though, to the potential ‘downstream’ implications of the ramping up of supply and the introduction of new technologies for build quality and defects. This paper contributes to our understanding of the ‘downstream’ implications through a synthesis of the current UK defect literature with respect to new-build housing. It is found that the prevailing emphasis in the literature is limited to the responsibility, pathology and statistical analysis of defects (and failures). The literature does not extend to how house builders individually and collectively, in practice, collect and learn from defects information. The paper concludes by describing an ongoing collaborative research programme with the National House Building Council (NHBC) to: (a) understand house builders’ localised defects analysis procedures, and their current knowledge feedback loops to inform risk management strategies; and, (b) building on this understanding, design and test action research interventions to develop new data capture, learning processes and systems to reduce targeted defects.

A high-density immunoblotting methodology for quantification of total protein levels and phosphorylation modifications

The components of many signaling pathways have been identified and there is now a need to conduct quantitative data-rich temporal experiments for systems biology and modeling approaches to better understand pathway dynamics and regulation. Here we present a modified Western blotting method that allows the rapid and reproducible quantification and analysis of hundreds of data points per day on proteins and their phosphorylation state at individual sites. The approach is of particular use where samples show a high degree of sample-to-sample variability such as primary cells from multiple donors. We present a case study on the analysis of >800 phosphorylation data points from three phosphorylation sites in three signaling proteins over multiple time points from platelets isolated from ten donors, demonstrating the technique's potential to determine kinetic and regulatory information from limited cell numbers and to investigate signaling variation within a population. We envisage the approach being of use in the analysis of many cellular processes such as signaling pathway dynamics to identify regulatory feedback loops and the investigation of potential drug/inhibitor responses, using primary cells and tissues, to generate information about how a cell's physiological state changes over time.

From insect to man: Photorhabdus sheds light on the emergence of human pathogenicity

Photorhabdus are highly effective insect pathogenic bacteria that exist in a mutualistic relationship with Heterorhabditid nematodes. Unlike other members of the genus, Photorhabdus asymbiotica can also infect humans. Most Photorhabdus cannot replicate above 34°C, limiting their host-range to poikilothermic invertebrates. In contrast, P. asymbiotica must necessarily be able to replicate at 37°C or above. Many well-studied mammalian pathogens use the elevated temperature of their host as a signal to regulate the necessary changes in gene expression required for infection. Here we use RNA-seq, proteomics and phenotype microarrays to examine temperature dependent differences in transcription, translation and phenotype of P. asymbiotica at 28°C versus 37°C, relevant to the insect or human hosts respectively. Our findings reveal relatively few temperature dependant differences in gene expression. There is however a striking difference in metabolism at 37°C, with a significant reduction in the range of carbon and nitrogen sources that otherwise support respiration at 28°C. We propose that the key adaptation that enables P. asymbiotica to infect humans is to aggressively acquire amino acids, peptides and other nutrients from the human host, employing a so called “nutritional virulence” strategy. This would simultaneously cripple the host immune response while providing nutrients sufficient for reproduction. This might explain the severity of ulcerated lesions observed in clinical cases of Photorhabdosis. Furthermore, while P. asymbiotica can invade mammalian cells they must also resist immediate killing by humoral immunity components in serum. We observed an increase in the production of the insect Phenol-oxidase inhibitor Rhabduscin normally deployed to inhibit the melanisation immune cascade. Crucially we demonstrated this molecule also facilitates protection against killing by the alternative human complement pathway.

Avoiding the subject media, culture and the object

"This is a collection of inter-related essays on the postmillennial mediascape. Focusing on the neglected significance of the object within today's discourse networks, Avoiding the Subject extends the formal possibilities of cultural criticism by highlighting feedback loops between philosophy, technology, and politics. Students and teachers of visual culture, critical theory, cultural studies, film theory, and new media will find a wealth of ideas and insights in this fresh approach to the electronic environment."--BOOK JACKET.

Exercise, diet and skeletal muscle gene expression

Skeletal muscle, as a consequence of its mass and great capacity for altered metabolism, has a major impact on whole-body metabolic homeostasis and is capable of remarkable adaptation in response to various physiological stimuli, including exercise and dietary intervention. Exercise-induced increases in skeletal muscle mRNA levels of a number of genes have been reported, due to transcriptional activation and/or increased mRNA stability. The cellular adaptations to exercise training appear to be due to the cumulative effects of transient increases in gene transcription after repeated exercise bouts. The relative importance of transcriptional (mRNA synthesis) and translational (mRNA stability or translational efficiency) mechanisms for the training-induced increases in skeletal muscle protein abundance remains to be fully elucidated. Dietary manipulation, and the associated alterations in nutrient availability and hormone levels, can also modify skeletal muscle gene expression, although fewer studies have been reported. A major challenge is to understand how exercise and diet exert their effects on gene and protein expression in skeletal muscle. In relation to exercise, potential stimuli include stretch and muscle tension, the pattern of motor nerve activity and the resultant calcium transients, the energy charge of the cell and substrate availability, oxygen tension and circulating hormones. These are detected by various cellular signaling mechanisms, acting on a range of downstream targets and a wide range of putative transcription factors. A key goal in the years ahead is to identify how alterations at the level of gene expression are coupled to the changes in skeletal muscle phenotype. It is clear that gene expression, although representing a specific site of regulation, is only one step in a complex cascade from the initial stimulus to the final phenotypic adaptation and integrated physiological response.

Virulence determinants of infectious bursal disease virus

The very virulent (vv) pathotype of infectious bursal disease virus (IBDV) has spread rapidly throughout Europe, Asia, and the Middle East. Although Australia is currently unaffected, there remains the potential for incursion of an exotic isolate. The aim of this study was to identify putative virulence determinants of IBDV to facilitate the development of improved diagnostic assays for detection and characterisation of vvIBDV isolates. Sequencing of Indonesian vvIBDV Tasik94 revealed a unique substitution [ A�¨S222] in the hypervariable region (HVR) of viral protein (VP) VP2, which did not appear to impinge on virulence or antigenicity. Phylogenetic analyses indicated that Tasik94 was closely related to Asian and European vvIBDV strains. Extensive alignment of deduced protein sequences across the HVR of VP2 identified residuesI242 I256 and I294 as putative markers of the vv phcnotype. Comparison of the pathology induced by mildly-virulent Australian IBDV 002/73 and Indonesian vvIBDV Tasik94, revealed that histological lesions in the spleen, thymus and bone marrow were restricted to Tasik94-infected birds, suggesting the enhanced pathogenicity of vvIBDV might be attributed to replication in non-bursal lymphoid organs. The biological significance of the VP2 HVR in virulence was assessed using recombinant viruses generated by reverse genetics. Both genomic segments of Australian IBDV 002/73, and recombinant segment A constructs in which the HVR of 002/73 was replaced with the corresponding region of either tissue culture-adapted virus or vvIBDV (Tasik94), were cloned behind T7 RNA polymerase promoter sequences. In vitro transcription/translation of each construct resulted in expression of viral proteins. Co-transfection of synthetic RNA transcripts initiated replication of both tissue culture-adapted parental and recombinant viruses, however attempts to rescue non-adapted viruses in specific-pathogen-free (SPF) chickens were unsuccessful. Nucleotide sequence variation in the HVR of VP2 was exploited for the development of a new diagnostic assay to rapidly detect exotic IBDV isolates, including vvIBDV, using reverse transcription polymerase chain reaction (RT-PCR) amplification and Bmrl restriction enzyme digestion. The assay was capable of differentiating between endemic and exotic IBDV in 96% of 105 isolates sequenced to date.

Action research of cyclonic transactions in online management education

The aim of the research was to generate a cyclonic model for understanding the influences and processes of continuously improving management education in an environment rich in online learning technologies. The research questions were:
1. What is the nature of the cyclonic interactions observed in the transactions of a team of online management educators?
2. How might an understanding of cyclonic interactions
a. help refine action research, and
b. generate rich insight for online management education?
The methodology was an action research project. The research team worked in an online Master of Business Administration (MBA) to continuously develop teaching practice in one unit of the MBA. The methodology matched the objectives of the project, and the appropriate rigour associated with qualitative, interpretive research. The results showed that theories of systems and relational dynamics, adapted to hermeneutics and aligned with other learning theories, can be framed by the metaphor of a cyclone to conduct research into teaching practice and build upon the theory base in the field of online education.
Online management education is subject to reinterpretations. The cyclonic framework explains some of the changes. The project showed that a chaotic but organised cyclonic program development process in one particular MBA course was informative for and informed by the chaotic and cyclonic globalized business world. For the education of managers the cyclonic view was relevant. The approach was metaphorical and, therefore, opened new ways of seeing and speaking. Findings pertained to the nature of the cyclonic interactions, how an understanding of cyclonic interactions helped to refine action research, and how an understanding of cyclonic interactions helped generate rich insight for online management education.
It was found that it was the asymmetrical impetus of imperfection that created the examples of cyclonic learning spirals formed as double feedback loops for improved understanding. Online education in the action research required cyclical enhancement of connectedness by teachers, stronger emphasis on relational considerations in learning, and heightened expectations of collaboration by educators. It became possible to correlate earlier conceptions of action research with cyclonic categories and analyse the parallels with events in this action research project. Models were developed and presented to explain 3 cyclonic connections with hermeneutics, collaborative teaching, online resource
development, and the environment of online management.

Depression and sickness behavior are Janus-faced responses to shared inflammatory pathways

It is of considerable translational importance whether depression is a form or a consequence of sickness behavior. Sickness behavior is a behavioral complex induced by infections and immune trauma and mediated by pro-inflammatory cytokines. It is an adaptive response that enhances recovery by conserving energy to combat acute inflammation. There are considerable phenomenological similarities between sickness behavior and depression, for example, behavioral inhibition, anorexia and weight loss, and melancholic (anhedonia), physio-somatic (fatigue, hyperalgesia, malaise), anxiety and neurocognitive symptoms. In clinical depression, however, a transition occurs to sensitization of immuno-inflammatory pathways, progressive damage by oxidative and nitrosative stress to lipids, proteins, and DNA, and autoimmune responses directed against self-epitopes. The latter mechanisms are the substrate of a neuroprogressive process, whereby multiple depressive episodes cause neural tissue damage and consequent functional and cognitive sequelae. Thus, shared immuno-inflammatory pathways underpin the physiology of sickness behavior and the pathophysiology of clinical depression explaining their partially overlapping phenomenology. Inflammation may provoke a Janus-faced response with a good, acute side, generating protective inflammation through sickness behavior and a bad, chronic side, for example, clinical depression, a lifelong disorder with positive feedback loops between (neuro)inflammation and (neuro)degenerative processes following less well defined triggers.

Mobile phone-based audience response system and student engagement in large-group teaching

Engaging students in large classes is a challenge at the best of times. Teachers are increasingly seeking the help of new technology to keep the attention of their technologically savvy students. VotApedia, a free cell phone-based audience response system, is one such technology. This research aims to assess economics students' perceptions of use of VotApedia in helping to achieve deeper learning; to map importance of use of audience response technology for students in promoting engagement and to assess the promotion of a positive and active environment in lectures by use of VotApedia in an Australian University. Using the framework of activity theory, it argues that technologies such as VotApedia use positive feedback loops to facilitate improved student engagement and learning.

Paradigm shift to enhanced water supply planning through augmented grids, scarcity pricing and adaptive factory water : a system dynamics approach

This paper details a system dynamics model developed to simulate proposed changes to water governance through the integration of supply, demand and asset management processes. To effectively accomplish this, interconnected feedback loops in tariff structures, demand levels and financing capacity are included in the model design, representing the first comprehensive life-cycle modelling of potable water systems. A number of scenarios were applied to Australia's populated South-east Queensland region, demonstrating that introducing temporary drought pricing (i.e. progressive water prices set inverse with availability), in conjunction with supply augmentation through rain-independent sources, is capable of efficiently providing water security in the future. Modelling demonstrated that this alternative tariff structure reduced demand in scarcity periods thereby preserving supply, whilst revenues are maintained to build new water supply infrastructure. In addition to exploring alternative tariffs, the potential benefits of using adaptive pressure-retarded osmosis desalination plants for both potable water and power generation was explored. This operation of these plants for power production, when they would otherwise be idle, shows promise in reducing their net energy and carbon footprints. Stakeholders in industry, government and academia were engaged in model development and validation. The constructed model displays how water resource systems can be reorganised to cope with systemic change and uncertainty.

Drought and desalination: Melbourne water supply and development choices in the twenty-first century

Sharply reduced catchment inflows across Australia around the end of the twentieth century led to a sequence of water restrictions followed, as the drought persisted, by approximately $10 billion of investments in desalination plants near Perth, Adelaide, Melbourne, Sydney and Brisbane. This Deakin University project jointly with Griffith University, for the National Centre of Excellence in Desalination (NCEDA), follows these new investments. We ask how best to manage bulk water supply and retail supply given the facts and fears of uncertain rainfall, modelled over a 100 year simulation period. We use Monte Carlo style studies aiming to capture the new tensions and trade-offs regarding uncertain climate, rainfall and water supply. There are presently no comprehensive life-cycle approaches to model city water balances that incorporate economic feedbacks, such as tariff adjustment, which can in turn create a financing capacity for such investment responses to low catchment levels, models that could provide significant policy implications for water planners. This project addresses the gap, and presents excerpts from a system dynamics model that augments the usual water utility representation of the physical linkages and water grids. We add inter-connected feedback loops in tariff structures, demand levels and financing capacity. Tariffs are reset in association with drought and the modelling of responses both in terms of reduced consumption and increased revenue to the utility, depending on the elasticities of demand responses to higher tariffs, both short and long term, while also allowing effects from any transitional restrictions. Before reporting on parts of the simulations applied to Melbourne, this paper will first review the general issues surrounding whether desalination is or can be a “game changer” for economic development that hinges on secure water supply. We then explore options in bulk water supply management when desalination augments the choices, including catchments, dams, recycling, pipelines from rivers and savings in irrigation. Finally, the paper addresses the intriguing and important question of the value and cost of providing water for environmental uses.

Water security through scarcity pricing and reverse osmosis: a system dynamics approach

Water supply and demand planning is often conducted independently of social and economic strategies. There are presently no comprehensive life-cycle approaches to modelling urban water balances that incorporate economic feedbacks, such as tariff adjustment, which can in turn create a financing capacity for investment responses to low reservoir levels. This paper addresses this gap, and presents a system dynamics model that augments the usual water utility representation of the physical linkages of water grids, by adding inter-connected feedback loops in tariff structures, demand levels and financing capacity. The model, applied in the south-east Queensland region in Australia, enables simulation of alternatives and analysis of stocks and flows around a grid or portfolio of bulk supplies including an increasing proportion of rain-independent desalination plants. Such rain-independent water production plants complement the rain-dependent sources in the region and can potentially offer indefinite water security at a price. The study also shows how an alternative temporary drought pricing regime not only defers costly bulk supply infrastructure but actually generates greater price stability than traditional pricing approaches. The model has implications for water supply planners seeking to pro-actively plan, justify and finance portfolios of rain-dependent and rain-independent bulk water supply infrastructure. Interestingly, the modelling showed that a temporary drought pricing regime not only lowers the frequency and severity of water insecurity events but also reduces the long-run marginal cost of water supply for the region when compared to traditional reactive planning approaches that focus on restrictions to affect demand in scarcity periods.

The best are never normal : exploring the distribution of firm performance

Competitive Strategy literature predicts three different mechanisms of performance generation, thus distinguishing between firms that have competitive advantage, firms that have competitive disadvantage or firms that have neither. Nonetheless, previous works in the field have fitted a single normal distribution to model firm performance. Here, we develop a new approach that distinguishes among performance generating mechanisms and allows the identification of firms with competitive advantage or disadvantage. Theorizing on the positive feedback loops by which firms with competitive advantage have facilitated access to acquire new resources, we proposed a distribution we believe data on firm performance should follow. We illustrate our model by assessing its fit to data on firm performance, addressing its theoretical implications and comparing it to previous works.

Citizens vs. banks: institutional drivers of financial market litigiousness in Brazil

This article analyzes the institutional drivers of Brazil’s alarmingly high levels of litigation between clients and financial institutions. Most of the policy oriented literature that explores that phenomenon discusses the impacts of a perceived debtor-friendly bias of Brazilian courts on generating feedback loops of litigation that further increases interest rates and creates adverse selection within the pool of potential debtors. This literature therefore addresses the way courts behave once disputes reach their doorstep; conversely, we take a step back to understand the underlying reasons for why such a large number of disputes end up in courts in the first place. We accordingly attribute endemic litigation in Brazilian financial markets to a framework of political, economic and legal institutions and circumstances, which this article aims to unbound and explain.