Use of synthetic CaV2.2 Ca2+ channel peptides to study presynaptic function

Small, synthetic peptides based on specific regions of voltage-gated Ca2+ channels (VGCCs) have been widely used to study Ca2+ channel function and have been instrumental in confirming the contribution of specific amino acid sequences to interactions with putative binding partners. In particular, peptides based on the Ca2+ channel Alpha Interaction Domain (AID) on the intracellular region connecting domains I and II (the I-II loop) and the SYNaptic PRotein INTerction (synprint) site on the II-III loop have been widely used. Emerging evidence suggests that such peptides may themselves possess inherent functionality, a property that may be exploitable for future drug design. Here, we review our recent work using synthetic Ca2+ channel peptides based on sequences within the CaV2.2 amino terminal and I-II loop, originally identified as molecular determinates for G protein modulation, and their effects on VGCC function. These CaV2.2 peptides act as inhibitory modules to decrease Ca2+ influx with direct effects on VGCC gating, ultimately leading to a reduction of synaptic transmission. CaV2.2 peptides also attenuate G protein modulation of VGCCs. Amino acid substitutions generate CaV2.2 peptides with increased or decreased inhibitory effects suggesting that synthetic peptides can be used to further probe VGCC function and, potentially, form the basis for novel therapeutic development.

Estimation of wind storm impacts over Western Germany under future climate conditions using a statistical-dynamical downscaling approach

A statistical–dynamical regionalization approach is developed to assess possible changes in wind storm impacts. The method is applied to North Rhine-Westphalia (Western Germany) using the FOOT3DK mesoscale model for dynamical downscaling and ECHAM5/OM1 global circulation model climate projections. The method first classifies typical weather developments within the reanalysis period using K-means cluster algorithm. Most historical wind storms are associated with four weather developments (primary storm-clusters). Mesoscale simulations are performed for representative elements for all clusters to derive regional wind climatology. Additionally, 28 historical storms affecting Western Germany are simulated. Empirical functions are estimated to relate wind gust fields and insured losses. Transient ECHAM5/OM1 simulations show an enhanced frequency of primary storm-clusters and storms for 2060–2100 compared to 1960–2000. Accordingly, wind gusts increase over Western Germany, reaching locally +5% for 98th wind gust percentiles (A2-scenario). Consequently, storm losses are expected to increase substantially (+8% for A1B-scenario, +19% for A2-scenario). Regional patterns show larger changes over north-eastern parts of North Rhine-Westphalia than for western parts. For storms with return periods above 20 yr, loss expectations for Germany may increase by a factor of 2. These results document the method's functionality to assess future changes in loss potentials in regional terms.

Assessment of the Wind Gust Estimate Method in mesoscale modelling of storm events over West Germany

A physically based gust parameterisation is added to the atmospheric mesoscale model FOOT3DK to estimate wind gusts associated with storms over West Germany. The gust parameterisation follows the Wind Gust Estimate (WGE) method and its functionality is verified in this study. The method assumes that gusts occurring at the surface are induced by turbulent eddies in the planetary boundary layer, deflecting air parcels from higher levels down to the surface under suitable conditions. Model simulations are performed with horizontal resolutions of 20 km and 5 km. Ten historical storm events of different characteristics and intensities are chosen in order to include a wide range of typical storms affecting Central Europe. All simulated storms occurred between 1990 and 1998. The accuracy of the method is assessed objectively by validating the simulated wind gusts against data from 16 synoptic stations by means of “quality parameters”. Concerning these parameters, the temporal and spatial evolution of the simulated gusts is well reproduced. Simulated values for low altitude stations agree particularly well with the measured gusts. For orographically exposed locations, the gust speeds are partly underestimated. The absolute maximum gusts lie in most cases within the bounding interval given by the WGE method. Focussing on individual storms, the performance of the method is better for intense and large storms than for weaker ones. Particularly for weaker storms, the gusts are typically overestimated. The results for the sample of ten storms document that the method is generally applicable with the mesoscale model FOOT3DK for mid-latitude winter storms, even in areas with complex orography.

A digital ecosystems model of assessment feedback on student learning

The term ecosystem has been used to describe complex interactions between living organisms and the physical world. The principles underlying ecosystems can also be applied to complex human interactions in the digital world. As internet technologies make an increasing contribution to teaching and learning practice in higher education, the principles of digital ecosystems may help us understand how to maximise technology to benefit active, self-regulated learning especially among groups of learners. Here, feedback on student learning is presented within a conceptual digital ecosystems model of learning. Additionally, we have developed a Web 2.0-based system, called ASSET, which incorporates multimedia and social networking features to deliver assessment feedback within the functionality of the digital ecosystems model. Both the digital ecosystems model and the ASSET system are described and their implications for enhancing feedback on student learning are discussed.

Wearable device to assist independent living

Older people increasingly want to remain living independently in their own homes. The aim of the ENABLE project is to develop a wearable device that can be used to support older people in their daily lives and which can monitor their health status, detect potential problems, provide activity reminders and offer communication and alarm services. In order to determine the specifications and functionality required for the development of the device, user surveys and focus groups were undertaken, use case analysis and scenario modeling carried out. The project has resulted in the development of a wrist-worn device and mobile phone combination that can support and assist older and vulnerable wearers with a range of activities and services both inside their home and as they move around their local environment. The device is currently undergoing pilot trials in five European countries. The aim of this paper is to describe the ENABLE device, its features and services, and the infrastructure within which it operates.

Optimal frame length for keeping normalized goodput with lowest requirement on BER

This paper presents an adaptive frame length mechanism based on a cross-layer analysis of intrinsic relations between the MAC frame length, bit error rate (BER) of the wireless link and normalized goodput. The proposed mechanism selects the optimal frame length that keeps the service normalized goodput at required levels while satisfying the lowest requirement on the BER, thus increasing the transmission reliability. Numerical results are provided and show that an optimal frame length satisfying the lowest BER requirement does indeed exist. The performance of BER requirement as a function of the MAC frame length is evaluated and compared for transmission scenarios with and without automatic repeat request (ARQ). Furthermore, issues related to the MAC overhead length are also discussed to illuminate the functionality and performance of the proposed mechanism.

Human microbiota in health and disease

Each human body plays host to a microbial population which is both numerically vast (at around 1014 microbial cells) and phenomenally diverse (over 1,000 species). The majority of the microbial species in the gut have not been cultured but the application of culture-independent approaches for high throughput diversity and functionality analysis has allowed characterisation of the diverse microbial phylotypes present in health and disease. Studies in monozygotic twins, showing that these retain highly similar microbiota decades after birth and initial colonisation, are strongly indicative that diversity of the microbiome is host-specific and affected by the genotype. Microbial diversity in the human body is reflected in both richness and evenness. Diversity increases steeply from birth reaching its highest point in early adulthood, before declining in older age. However, in healthy subjects there appears to be a core of microbial phylotypes which remains relatively stable over time. Studies of individuals from diverse geopraphies suggest that clusters of intestinal bacterial groups tend to occur together, constituting ‘enterotypes’. So variation in intestinal microbiota is stratified rather than continuous and there may be a limited number of host/microbial states which respond differently to environmental influences. Exploration of enterotypes and functional groups may provide biomarkers for disease and insights into the potential for new treatments based on manipulation of the microbiome. In health, the microbiota interact with host defences and exist in harmonious homeostasis which can then be disturbed by invading organisms or when ‘carpet bombing’ by antibiotics occurs. In a portion of individuals with infections, the disease will resolve itself without the need for antibiotics and microbial homeostasis with the host’s defences is restored. The administration of probiotics (live microorganisms which when administered in adequate amounts confer a health benefit on the host) represents an artificial way to enhance or stimulate these natural processes. The study of innate mechanisms of antimicrobial defence on the skin, including the production of numerous antimicrobial peptides (AMPs), has shown an important role for skin commensal organisms. These organisms may produce AMPs, and also amplify the innate immune responses to pathogens by activating signalling pathways and processing host produced AMPs. Research continues into how to enhance and manipulate the role of commensal organisms on the skin. The challenges of skin infection (including diseases caused by multiply resistant organisms) and infestations remain considerable. The potential to re-colonise the skin to replace or reduce pathogens, and exploring the relationship between microbiota elsewhere and skin diseases are among a growing list of research targets. Lactobacillus species are among the best known ‘beneficial’ bacterial members of the human microbiota. Of the approximately 120 species known, about 15 are known to occur in the human vagina. These organisms have multiple properties, including the production of lactic acid, hydrogen peroxide and bacteriocins, which render the vagina inhospitable to potential pathogens. Depletion of the of the normal Lactobacillus population and overgrowth of vaginal anaerobes, accompanied by the loss of normal vaginal acidity can lead to bacterial vaginosis – the commonest cause of abnormal vaginal discharge in women. Some vaginal anaerobes are associated with the formation of vaginal biofilms which serve to act as a reservoir of organisms which persists after standard antibiotic therapy of bacterial vaginosis and may help to account for the characteristically high relapse rate in the condition. Administration of Lactobacillus species both vaginally and orally have shown beneficial effects in the treatment of bacterial vaginosis and such treatments have an excellent overall safety record. Candida albicans is a frequent coloniser of human skin and mucosal membranes, and is a normal part of the microbiota in the mouth, gut and vagina. Nevertheless Candida albicans is the most common fungal pathogen worldwide and is a leading cause of serious and often fatal nosocomial infections. What turns this organism from a commensal to a pathogen is a combination of increasing virulence in the organism and predisposing host factors that compromise immunity. There has been considerable research into the use of probiotic Lactobacillus spp. in vaginal candidiasis. Studies in reconstituted human epithelium and monolayer cell cultures have shown that L. rhamnosus GG can protect mucosa from damage caused by Candida albicans, and enhance the immune responses of mucosal surfaces. Such findings offer the promise that the use of such probiotic bacteria could provide new options for antifungal therapy. Studies of changes of the human intestinal microbiota in health and disease are complicated by its size and diversity. The Alimentary Pharmabiotic Centre in Cork (Republic of Ireland) has the mission to ‘mine microbes for mankind’ and its work illustrates the potential benefits of understanding the gut microbiota. Work undertaken at the centre includes: mapping changes in the microbiota with age; studies of the interaction between the microbiota and the gut; potential interactions between the gut microbiota and the central nervous system; the potential for probiotics to act as anti-infectives including through the production of bacteriocins; and the characterisation of interactions between gut microbiota and bile acids which have important roles as signalling molecules and in immunity. The important disease entity where the role of the gut microbiota appears to be central is the Irritable Bowel Syndrome (IBS). IBS patients show evidence of immune activation, impaired gut barrier function and abnormal gut microbiota. Studies with probiotics have shown that these organisms can exert anti-inflammatory effects in inflammatory bowel disease and may strengthen the gut barrier in IBS of the diarrhoea-predominant type. Formal randomised trials of probiotics in IBS show mixed results with limited benefit for some but not all. Studies confirm that administered probiotics can survive and temporarily colonise the gut. They can also stimulate the numbers of other lactic acid bacilli in the gut, and reduce the numbers of pathogens. However consuming live organisms is not the only way to influence gut microbiota. Dietary prebiotics are selectively fermented ingredients that can change the composition and/or activity of the gastrointestinal microbiota in beneficial ways. Dietary components that reach the colon, and are available to influence the microbiota include poorly digestible carbohydrates, such as non-starch polysaccharides, resistant starch, non-digestible oligosaccharides (NDOs) and polyphenols. Mixtures of probiotic and prebiotic ingredients that can selectively stimulate growth or activity of health promoting bacteria have been termed ‘synbiotics’. All of these approaches can influence gut microbial ecology, mainly to increase bifidobacteria and lactobacilli, but metagenomic approaches may reveal wider effects. Characterising how these changes produce physiological benefits may enable broader use of these tactics in health and disease in the future. The current status of probiotic products commercially available worldwide is less than ideal. Prevalent problems include misidentification of ingredient organisms and poor viability of probiotic microorganisms leading to inadequate shelf life. On occasions these problems mean that some commercially available products cannot be considered to meet the definition of a probiotic product. Given the potential benefits of manipulating the human microbiota for beneficial effects, there is a clear need for improved regulation of probiotics. The potential importance of the human microbiota cannot be overstated. ‘We feed our microbes, they talk to us and we benefit. We just have to understand and then exploit this.’ (Willem de Vos).

Patterning and detailed study of human hNT astrocytes on parylene-C/silicon dioxide substrates to the single cell level

It is estimated that the adult human brain contains 100 billion neurons with 5–10 times as many astrocytes. Although it has been generally considered that the astrocyte is a simple supportive cell to the neuron, recent research has revealed new functionality of the astrocyte in the form of information transfer to neurons of the brain. In our previous work we developed a protocol to pattern the hNT neuron (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/SiO2 substrates. In this work, we report how we have managed to pattern hNT astrocytes, on parylene-C/SiO2 substrates to single cell resolution. This article disseminates the nanofabrication and cell culturing steps necessary for the patterning of such cells. In addition, it reports the necessary strip lengths and strip width dimensions of parylene-C that encourage high degrees of cellular coverage and single cell isolation for this cell type. The significance in patterning the hNT astrocyte on silicon chip is that it will help enable single cell and network studies into the undiscovered functionality of this interesting cell, thus, contributing to closer pathological studies of the human brain.

Expression of epitope-tagged LMW glutenin subunits in the starchy endosperm of transgenic wheat and their incorporation into glutenin polymers

The low-molecular-weight (LMW) glutenin subunits are components of the highly cross-linked glutenin polymers that confer viscoelastic properties to gluten and dough. They have both quantitative and qualitative effects on dough quality that may relate to differences in their ability to form the inter-chain disulphide bonds that stabilise the polymers. In order to determine the relationship between dough quality and the amounts and properties of the LMW subunits, we have transformed the pasta wheat cultivars Svevo and Ofanto with three genes encoding proteins, which differ in their numbers or positions of cysteine residues. The transgenes were delivered under control of the high-molecular-weight (HMW) subunit 1Dx5 gene promoter and terminator regions, and the encoded proteins were C-terminally tagged by the introduction of the c-myc epitope. Stable transformants were obtained with both cultivars, and the use of a specific antibody to the c-myc epitope tag allowed the transgene products to be readily detected in the complex mixture of LMW subunits. A range of transgene expression levels was observed. The addition of the epitope tag did not compromise the correct folding of the trangenic subunits and their incorporation into the glutenin polymers. Our results demonstrate that the ability to specifically epitope-tag LMW glutenin transgenes can greatly assist in the elucidation of their individual contributions to the functionality of the complex gluten system.

Early modern Oxford bindings in twenty-first century markup

Purpose – The Bodleian Binders Book contains nearly 150 pages of seventeenth century library records, revealing information about the binders used by the library and the thousands of bindings they produced. The purpose of this paper is to explore a pilot project to survey and record bindings information contained in the Binders Book. Design/methodology/approach – A sample size of seven pages (91 works, 65 identifiable bindings) to develop a methodology for surveying and recording bindings listed in the manuscript. To create a successful product that would be useful to bindings researchers, it addressed questions of bindings terminology and the role of the library in the knowledge creation process within the context that text encoding is changing the landscape of library functions. Text encoding formats were examined, and a basic TEI (Text Encoding Initiative) transcription was produced. This facilitates tagging of names and titles and the display of transcriptions with text images. Findings – Encoding was found not only to make the manuscript content more accessible, but to allow for the construction of new knowledge: characteristic Oxford binding traits were revealed and bindings were matched to binders. Plans for added functionality were formed. Originality/value – This research presents a “big picture” analysis of Oxford bindings as a result of text encoding and the foundation for qualitative and statistical analysis. It exemplifies the benefits of interdisciplinary methods – in this case from Digital Humanities – to enhance access to and interpretation of specialist materials and the library's provenance record.

The consumer acceptance of novel vegetable-enriched bread products as a potential vehicle to increase vegetable consumption

Currently UK fruit and vegetable intakes are below recommendations. Bread is a staple food consumed by ~95% of adults in western countries. In addition, bread provides an ideal matrix by which functionality can be delivered to the consumer in an accepted food. Therefore, enriching bread with vegetables may be an effective strategy to increase vegetable consumption. This study evaluated consumer acceptance, purchase intent and intention of product replacement of bread enriched with red beetroot, carrot with coriander, red pepper with tomato or white beetroot (80g vegetable per serving of 200g) compared to white control bread (0g vegetable). Consumers (n=120) rated their liking of the breads overall, as well as their liking of appearance, flavour and texture using nine-point hedonic scales. Product replacement and purchase intent of the breads was rated using five-point scales. The effect of providing consumers with health information about the breads was also evaluated. There were significant differences in overall liking (P<0.0001), as well as liking of appearance (P<0.0001), flavour (P=0.0002) and texture (P=0.04), between the breads. However, the significant differences resulted from the red beetroot bread which was significantly (P<0.05) less liked compared to control bread. There were no significant differences in overall liking between any of the other vegetable-enriched breads compared with the control bread (no vegetable inclusion), apart from the red beetroot bread which was significantly less liked. The provision of health information about the breads did not increase consumer liking of the vegetable-enriched breads. In conclusion, this study demonstrated that vegetable-enriched bread appeared to be an acceptable strategy to increase vegetable intake, however, liking depended on vegetable type.

What is the right balance for performance and isolation with virtualization in HPC?

The use of virtualization in high-performance computing (HPC) has been suggested as a means to provide tailored services and added functionality that many users expect from full-featured Linux cluster environments. The use of virtual machines in HPC can offer several benefits, but maintaining performance is a crucial factor. In some instances the performance criteria are placed above the isolation properties. This selective relaxation of isolation for performance is an important characteristic when considering resilience for HPC environments that employ virtualization. In this paper we consider some of the factors associated with balancing performance and isolation in configurations that employ virtual machines. In this context, we propose a classification of errors based on the concept of “error zones”, as well as a detailed analysis of the trade-offs between resilience and performance based on the level of isolation provided by virtualization solutions. Finally, a set of experiments are performed using different virtualization solutions to elucidate the discussion.

Change management in practice: an ethnographic study of changes to contract requirements on a hospital project

Changes to client requirements are inevitable during construction. Industry discourse is concerned with minimizing and controlling changes. However, accounts of practices involved in making changes are rare. In response to calls for more research into working practices, an ethnographic study of a live hospital project was undertaken to explore how changes are made. A vignette of a meeting exploring the investigation of changes illustrates the issues. This represents an example from the ethnographic fieldwork, which produced many observations. There was a strong emphasis on using change management procedures contained within the contract to investigate changes, even when it was known that the change was not required. For the practitioners, this was a way of demonstrating best practice, transparent and accountable decision-making regarding changes. Hence, concerns for following procedures sometimes overshadowed considerations about whether or not a change was required to improve the functionality of the building. However, the procedures acted as boundary objects between the communities of practice involved on the project by coordinating the work of managing changes. Insights suggest how contract procedures facilitate and impede the making of changes, which can inform policy guidance and contract drafting.

Divergent evolution of the activity and regulation of the glutamate decarboxylase systems in Listeria monocytogenes EGD-e and 10403S : roles in virulence and acid tolerance

The glutamate decarboxylase (GAD) system has been shown to be important for the survival of Listeria monocytogenes in low pH environments. The bacterium can use this faculty to maintain pH homeostasis under acidic conditions. The accepted model for the GAD system proposes that the antiport of glutamate into the bacterial cell in exchange for γ-aminobutyric acid (GABA) is coupled to an intracellular decarboxylation reaction of glutamate into GABA that consumes protons and therefore facilitates pH homeostasis. Most strains of L. monocytogenes possess three decarboxylase genes (gadD1, D2 & D3) and two antiporter genes (gadT1 & gadT2). Here, we confirm that the gadD3 encodes a glutamate decarboxylase dedicated to the intracellular GAD system (GADi), which produces GABA from cytoplasmic glutamate in the absence of antiport activity. We also compare the functionality of the GAD system between two commonly studied reference strains, EGD-e and 10403S with differences in terms of acid resistance. Through functional genomics we show that EGD-e is unable to export GABA and relies exclusively in the GADi system, which is driven primarily by GadD3 in this strain. In contrast 10403S relies upon GadD2 to maintain both an intracellular and extracellular GAD system (GADi/GADe). Through experiments with a murinised variant of EGD-e (EGDm) in mice, we found that the GAD system plays a significant role in the overall virulence of this strain. Double mutants lacking either gadD1D3 or gadD2D3 of the GAD system displayed reduced acid tolerance and were significantly affected in their ability to cause infection following oral inoculation. Since EGDm exploits GADi but not GADe the results indicate that the GADi system makes a contribution to virulence within the mouse. Furthermore, we also provide evidence that there might be a separate line of evolution in the GAD system between two commonly used reference strains.

iPad use in fieldwork: formal and informal use to enhance pedagogical practice in a bring your own technology world

We report on use of iPads (and other IOS devices) for student fieldwork use and as electronic field notebooks and to promote active. We have used questionnaires and interviews of tutors and students to elicit their views and technology and iPad use for fieldwork. There is some reluctance for academic staff to relinquish paper notebooks for iPad use, whether in the classroom or on fieldwork, as well as use them for observational and measurement purposes. Students too are largely unaware of the potential of iPads for enhancing fieldwork. Apps can be configured for a wide variety of specific uses that make iPads useful for educational as well as social uses. Such abilities should be used to enhance existing practice as well as make new functionality. For example, for disabled students who find it difficult to use conventional note taking. iPads can be used to develop student self-directed learning and for group contributions. The technology becomes part of the students’ personal learning environments as well as at the heart of their knowledge spaces – academic and social. This blurring of boundaries is due to iPads’ usability to cultivate field use, instruction, assessment and feedback processes. iPads can become field microscopes and entries to citizen science and we see the iPad as the main ‘computing’ device for students in the near future. As part of the Bring Your Own Technology/Device (BYOD) the iPad has much to offer although, both staff and students need to be guided in the most effective use for self-directed education via development of Personal Learning Environments. A more student-oriented pedagogy is suggested to correspond to the increasing use of tablet technologies by students