Mastering the canonical loop of serine protease inhibitors : enhancing potency by optimising the internal hydrogen bond network

Background Canonical serine protease inhibitors commonly bind to their targets through a rigid loop stabilised by an internal hydrogen bond network and disulfide bond(s). The smallest of these is sunflower trypsin inhibitor (SFTI-1), a potent and broad-range protease inhibitor. Recently, we re-engineered the contact β-sheet of SFTI-1 to produce a selective inhibitor of kallikrein-related peptidase 4 (KLK4), a protease associated with prostate cancer progression. However, modifications in the binding loop to achieve specificity may compromise structural rigidity and prevent re-engineered inhibitors from reaching optimal binding affinity. Methodology/Principal Findings In this study, the effect of amino acid substitutions on the internal hydrogen bonding network of SFTI were investigated using an in silico screen of inhibitor variants in complex with KLK4 or trypsin. Substitutions favouring internal hydrogen bond formation directly correlated with increased potency of inhibition in vitro. This produced a second generation inhibitor (SFTI-FCQR Asn14) which displayed both a 125-fold increased capacity to inhibit KLK4 (Ki = 0.0386±0.0060 nM) and enhanced selectivity over off-target serine proteases. Further, SFTI-FCQR Asn14 was stable in cell culture and bioavailable in mice when administered by intraperitoneal perfusion. Conclusion/Significance These findings highlight the importance of conserving structural rigidity of the binding loop in addition to optimising protease/inhibitor contacts when re-engineering canonical serine protease inhibitors.

Enabling USDL by tools

Fundamental tooling is required in order to apply USDL in practical settings. This chapter discusses three fundamental types of tools for USDL. First, USDL editors have been developed for expert and casual users, respectively. Second, several USDL repositories have been built to allow editors accessing and storing USDL descriptions. Third, our generic USDL marketplace allows providers to describe their services once and potentially trade them anywhere. In addition, the iosyncrasies of service trading as opposed to the simpler case of product trading. The chapter also presents several deployment scenarios of such tools to foster individual value chains and support new business models across organizational boundaries. We close the chapter with an application of USDL in the context of service engineering.

SOA approaches

As the service-oriented architecture paradigm has become ever more popular, different standardization efforts have been proposed by various consortia to enable interaction among heterongeneous environments through this paradigm. This chapter will overview the most prevalent of these SOA approaches. It will first show how technical services can be described, how they can interact with each other and be discovered by users. Next, the chapter will present different standards to facilitate service composition and to design service-oriented environments in light of a universal understanding of service orientation. The chapter will conclude with a summary and a discussion on the limitations of the reviewed standards along their ability to describe service properties. This paves the way to the next chapters where the USDL standard will be presented, which aims to lift such limitations.

Design overview of USDL

Enabling web-based service networks and ecosystems requires a way of describing services by a "commercial envelope" as discussed in Chapter 1. A uniform conception of services across all walks of life (including technical services) is required capturing business, operational and technical aspects. Therefore, our proposed Unified Service Description Language (USDL) particularly draws from and generalizes the best-of-breed approaches presented in Part I. The following chapter presents the design rationale of USDL where the different aspects are put in a framework of descriptions requirements. This is followed by the subsequent chapters of this part that provide details on specific aspects such as pricing or legal issues.

Domestic energy monitoring in Victorian households with PV solar installations : feasibility report for GV Community Energy

Australia requires decisive action on climate change and issues of sustainability. The Urban Informatics Research Lab has been funded by the Queensland State Government to conduct a three year study (2009 – 2011) exploring ways to support Queensland residents in making more sustainable consumer and lifestyle choices. We conduct user-centred design research that inform the development of real-time, mobile, locational, networked information interfaces, feedback mechanisms and persuasive and motivational approaches that in turn assist in-situ decision making and environmental awareness in everyday settings. The study aims to deliver usable and useful prototypes offering individual and collective visualisations of ecological impact and opportunities for engagement and collaboration in order to foster a participatory and sustainable culture of life in Australia. Raising people’s awareness with environmental data and educational information does not necessarily trigger sufficient motivation to change their habits towards a more environmentally friendly and sustainable lifestyle. Our research seeks to develop a better understanding how to go beyond just informing and into motivating and encouraging action and change. Drawing on participatory culture, ubiquitous computing, and real-time information, the study delivers research that leads to viable new design approaches and information interfaces which will strengthen Australia’s position to meet the targets of the Clean Energy Future strategy, and contribute to the sustainability of a low-carbon future in Australia. As part of this program of research, the Urban Informatics Research Lab has been invited to partner with GV Community Energy Pty Ltd on a project funded by the Victorian Government Sustainability Fund. This feasibility report specifically looks at the challenges and opportunities of energy monitoring in households in Victoria that include a PV solar installation. The report is structured into two parts: In Part 1, we first review a range of energy monitoring solutions, both stand-alone and internet-enabled. This section primarily focusses on the technical capacilities. However, in order to understand this information and make an informed decision, it is crucial to understand the basic principles and limitations of energy monitoring as well as the opportunities and challenges of a networked approach towards energy monitoring which are discussed in Section 2.

Human proximal tubule epithelial cells modulate autologous dendritic cell function

Background We have previously demonstrated that human kidney proximal tubule epithelial cells (PTEC) are able to modulate autologous T and B lymphocyte responses. It is well established that dendritic cells (DC) are responsible for the initiation and direction of adaptive immune responses and that these cells occur in the renal interstitium in close apposition to PTEC under inflammatory disease settings. However, there is no information regarding the interaction of PTEC with DC in an autologous human context. Methods Human monocytes were differentiated into monocyte-derived DC (MoDC) in the absence or presence of primary autologous activated PTEC and matured with polyinosinic:polycytidylic acid [poly(I:C)], while purified, pre-formed myeloid blood DC (CD1c+ BDC) were cultured with autologous activated PTEC in the absence or presence of poly(I:C) stimulation. DC responses were monitored by surface antigen expression, cytokine secretion, antigen uptake capacity and allogeneic T-cell-stimulatory ability. Results The presence of autologous activated PTEC inhibited the differentiation of monocytes to MoDC. Furthermore, MoDC differentiated in the presence of PTEC displayed an immature surface phenotype, efficient phagocytic capacity and, upon poly(I:C) stimulation, secreted low levels of pro-inflammatory cytokine interleukin (IL)-12p70, high levels of anti-inflammatory cytokine IL-10 and induced weak Th1 responses. Similarly, pre-formed CD1c+ BDC matured in the presence of PTEC exhibited an immature tolerogenic surface phenotype, strong endocytic and phagocytic ability and stimulated significantly attenuated T-cell proliferative responses. Conclusions Our data suggest that activated PTEC regulate human autologous immunity via complex interactions with DC. The ability of PTEC to modulate autologous DC function has important implications for the dampening of pro-inflammatory immune responses within the tubulointerstitium in renal injuries. Further dissection of the mechanisms of PTEC modulation of autologous immune responses may offer targets for therapeutic intervention in renal medicine.

The α5 neuronal nicotinic acetylcholine receptor subunits plays an important role in the sedative effects of ethanol but does not modulate consumption in mice

Alcohol use disorders (AUDs) are a major public health problem, and the few treatment options available to those seeking treatment offer only modest success rates. There remains a need to identify novel targets for the treatment of AUDs. The neuronal nicotinic acetylcholine receptors (nAChRs) represent a potential therapeutic target in the brain, as recent human genetic studies have implicated gene variants in the α5 nAChR subunit as high risk factors for developing alcohol dependence. Here, we evaluate the role of 5* nAChR for ethanol-mediated behaviors using α5+/+ and α5-/- mice. We characterized the effect of hypnotic doses of ethanol and investigated drinking behavior using an adapted Drinking-in-the Dark (DID) paradigm that has been shown to induce high ethanol consumption in mice. We found the α5 subunit to be critical in mediating the sedative effects of ethanol. The α5-/- mice showed slower recovery from ethanol-induced sleep, as measured by loss of righting reflex. Additionally the α5-/- mice showed enhanced impairment to ethanol-induced ataxia. We found the initial sensitivity to ethanol and ethanol metabolism to be similar in both α5+/+ and α5-/- mice. Hence the enhanced sedation is likely due to a difference in the acute tolerance of ethanol in mice deficient of the α5 subunit. However the α5 subunit did not play a role in ethanol consumption for ethanol concentrations ranging from 5% to 30% in the DID paradigm. Additionally, varenicline (Chantix®) was effective in reducing ethanol intake in α5-/- mice. Together, our data suggest that the α5 nAChR subunit is important for the sedative hypnotic doses of ethanol but does not play a role in ethanol consumption. Varenicline can be a treatment option even when there is loss of function of the α5 nAChR subunit.

Levesys : innovation management enabling a conversation about the future

This case study reports on the impact and business transformation of an IMP³rove assessment and follow-up workshop on Australian SME LEVESYS (www.levesys.com), which was undertaken by QMI Solutions. Innovation was not a foreign term to the company, which focuses on the development of enterprise resource planning (ERP) software for the Australian construction sector. However, before seeing and undergoing the IMP³rove process, this company had difficulty articulating their innovation problems and, therefore, had not achieved growth targets from its R&D efforts. This case study highlights the role of IMP³rove in assisting LEVESYS to take the first step in transforming itself through innovation.

Fibroblast growth factor receptor inhibition synergizes with paclitaxel and doxorubicin in endometrial cancer cells

OBJECTIVE: The fibroblast growth factor (FGF) family of signaling molecules has been associated with chemoresistance and poor prognosis in a number of cancer types, including lung, breast, ovarian, prostate, and head and neck carcinomas. Given the identification of activating mutations in the FGF receptor 2 (FGFR2) receptor tyrosine kinase in a subset of endometrial tumors, agents with activity against FGFRs are currently being tested in clinical trials for recurrent and progressive endometrial cancer. Here, we evaluated the effect of FGFR inhibition on the in vitro efficacy of chemotherapy in endometrial cancer cell lines. METHODS: Human endometrial cancer cell lines with wild-type or activating FGFR2 mutations were used to determine any synergism with concurrent use of the pan-FGFR inhibitor, PD173074, and the chemotherapeutics, doxorubicin and paclitaxel, on cell proliferation and apoptosis. RESULTS: FGFR2 mutation status did not alter sensitivity to either chemotherapeutic agent alone. The combination of PD173074 with paclitaxel or doxorubicin showed synergistic activity in the 3 FGFR2 mutant cell lines evaluated. In addition, although nonmutant cell lines were resistant to FGFR inhibition alone, the addition of PD173074 potentiated the cytostatic effect of paclitaxel and doxorubicin in a subset of FGFR2 wild-type endometrial cancer cell lines. CONCLUSIONS: Together these data suggest a potential therapeutic benefit to combining an FGFR inhibitor with standard chemotherapeutic agents in endometrial cancer therapy particularly in patients with FGFR2 mutation positive tumors.

Visual servoing approach to collision avoidance for aircraft

This paper presents a reactive Sense and Avoid approach using spherical image-based visual servoing. Avoidance of point targets in the lateral or vertical plane is achieved without requiring an estimate of range. Simulated results for static and dynamic targets are provided using a realistic model of a small fixed wing unmanned aircraft.

Beyond barcoding : a mitochondrial genomics approach to molecular phylogenetics and diagnostics of blowflies (Diptera: Calliphoridae)

Members of the Calliphoridae (blowflies) are significant for medical and veterinary management, due to the ability of some species to consume living flesh as larvae, and for forensic investigations due to the ability of others to develop in corpses. Due to the difficulty of accurately identifying larval blowflies to species there is a need for DNA-based diagnostics for this family, however the widely used DNA-barcoding marker, cox1, has been shown to fail for several groups within this family. Additionally, many phylogenetic relationships within the Calliphoridae are still unresolved, particularly deeper level relationships. Sequencing whole mt genomes has been demonstrated both as an effective method for identifying the most informative diagnostic markers and for resolving phylogenetic relationships. Twenty-seven complete, or nearly so, mt genomes were sequenced representing 13 species, seven genera and four calliphorid subfamilies and a member of the related family Tachinidae. PCR and sequencing primers developed for sequencing one calliphorid species could be reused to sequence related species within the same superfamily with success rates ranging from 61% to 100%, demonstrating the speed and efficiency with which an mt genome dataset can be assembled. Comparison of molecular divergences for each of the 13 protein-coding genes and 2 ribosomal RNA genes, at a range of taxonomic scales identified novel targets for developing as diagnostic markers which were 117–200% more variable than the markers which have been used previously in calliphorids. Phylogenetic analysis of whole mt genome sequences resulted in much stronger support for family and subfamily-level relationships. The Calliphoridae are polyphyletic, with the Polleninae more closely related to the Tachinidae, and the Sarcophagidae are the sister group of the remaining calliphorids. Within the Calliphoridae, there was strong support for the monophyly of the Chrysomyinae and Luciliinae and for the sister-grouping of Luciliinae with Calliphorinae. Relationships within Chrysomya were not well resolved. Whole mt genome data, supported the previously demonstrated paraphyly of Lucilia cuprina with respect to L. sericata and allowed us to conclude that it is due to hybrid introgression prior to the last common ancestor of modern sericata populations, rather than due to recent hybridisation, nuclear pseudogenes or incomplete lineage sorting.

Investigating the deployment of project management : a new perspective based on the concept of certification

Purpose – The purpose of this paper is to provide a summary description of the doctoral thesis investigating the field of project management (PM) deployment. Researchers will be informed of the current contributions within this topic and of the possible further investigations and researches. The decision makers and practitioners will be aware of a set of tools addressing the PM deployment with new perspectives. Design/methodology/approach – Research undertaken with the thesis is based on quantitative methods using time series statistics (time distance analysis) and comparative and correlation analysis aimed to better define and understand the PM deployment within and between countries or groups. Findings – The results suggest a project management deployment index (PMDI) to objectively measure the PM deployment based on the concept of certification. A proposed framework to empirically benchmark the PM deployment between countries by integrating the PMDI time series with the two dimensional comparative analysis of Sicherl. The correlation analysis within Hoftsede cultural framework shows the impact of the national culture dimensions on the PM deployment. The forecasting model shows a general continual growth trend of the PM deployment, with continual increase in the time distance between the countries. Research limitations/implications – The PM researchers are offered an empirical quantification on which they can construct further investigations and understanding of this phenomenon. The number of possible units that can be studied offers wide possibilities to replicate the thesis work. New researches can be undertaken to investigate further the contribution of other social or economical indicators, or to refine and enrich the definition of the PMDI indicator. Practical implications – These results have important implications on the PM deployment approaches. The PMDI measurements and time series comparisons facilitate considerably the measurement and benchmarking between the units (e.g. countries) and against targets, while the readiness setting of the studied unit (in terms of development and cultural levels) impacts the PM deployment within this country. Originality/value – This paper provides a summary of cutting-edge research work in the studied field of PM deployment and a link to the published works that researchers can use to help them understand the thesis research as well as how it can be extended.