The duration of Chlamydia muridarum genital tract infection and associated chronic pathological changes are reduced in IL-17 knockout mice but protection is not increased further by immunization

IL-17 is believed to be important for protection against extracellular pathogens, where clearance is dependent on neutrophil recruitment and local activation of epithelial cell defences. However, the role of IL-17 in protection against intracellular pathogens such as Chlamydia is less clear. We have compared (i) the course of natural genital tract C. muridarum infection, (ii) the development of oviduct pathology and (iii) the development of vaccine-induced immunity against infection in wild type (WT) BALB/c and IL-17 knockout mice (IL-17-/-) to determine if IL-17-mediated immunity is implicated in the development of infection-induced pathology and/or protection. Both the magnitude and duration of genital infection was significantly reduced in IL-17-/- mice compared to BALB/c. Similarly, hydrosalpinx was also greatly reduced in IL-17-/- mice and this correlated with reduced neutrophil and macrophage infiltration of oviduct tissues. Matrix metalloproteinase (MMP) 9 and MMP2 were increased in WT oviducts compared to IL-17-/- animals at day 7 post-infection. In contrast, oviducts from IL-17-/- mice contained higher MMP9 and MMP2 at day 21. Infection also elicited higher levels of Chlamydia-neutralizing antibody in serum of IL-17-/- mice than WT mice. Following intranasal immunization with C. muridarum Major Outer Membrane Protein (MOMP) and cholera toxin plus CpG adjuvants, significantly higher levels of chlamydial MOMP-specific IgG and IgA were found in serum and vaginal washes of IL-17-/- mice. T cell proliferation and IFNγ production by splenocytes was greater in WT animals following in vitro re-stimulation, however vaccination was only effective at reducing infection in WT, not IL-17-/- mice. Intranasal or transcutaneous immunization protected WT but not IL-17-/- mice against hydrosalpinx development. Our data show that in the absence of IL-17, the severity of C. muridarum genital infection and associated oviduct pathology are significantly attenuated, however neither infection or pathology can be reduced further by vaccination protocols that effectively protect WT mice.

Mapping local solutions to entrenched transport problems: Key lessons regarding the use of geographical information technologies in community mapping with disadvantaged communities

There is an increasing interest in the use of information technology as a participatory planning tool, particularly the use of geographical information technologies to support collaborative activities such as community mapping. However, despite their promise, the introduction of such technologies does not necessarily promote better participation nor improve collaboration. In part this can be attributed to a tendency for planners to focus on the technical considerations associated with these technologies at the expense of broader participation considerations. In this paper we draw on the experiences of a community mapping project with disadvantaged communities in suburban Australia to highlight the importance of selecting tools and techniques which support and enhance participatory planning. This community mapping project, designed to identify and document community-generated transport issues and solutions, had originally intended to use cadastral maps extracted from the government’s digital cadastral database as the foundation for its community mapping approach. It was quickly discovered that the local residents found the cadastral maps confusing as the maps lacked sufficient detail to orient them to their suburb (the study area). In response to these concerns and consistent with the project’s participatory framework, a conceptual base map based on resident’s views of landmarks of local importance was developed to support the community mapping process. Based on this community mapping experience we outline four key lessons learned regarding the process of community mapping and the place of geographical information technologies within this process.

Design LED innovation to rejuvenate local food systems and healthy communities : an emerging research agenda

Food Sovereignty (food freedom) is about empowering people to develop their own local food system. Food Sovereignty challenges designers to enable people to innovate the local food system, rather than having a food system which is dictated by corporate interests and failed business ethics. Communities are realising the potential for design to assist in the innovation process, and add strategic value to potentially localise the food system. Design Led Innovation (DLI) offers a strategic framework to address large-scale cultural, systemic and economic changes. The DLI approach empowers communities to take organised action to achieve a healthy, prosperous and happy way of life. DLI can assist with business models in the business world and it is evident this approach can assist with creating social change too. This paper presents on an emerging research agenda aimed to assist designer’s focus from individuals and systems to communities and urban problems. This paper also presents the research proposition that DLI and service design coupled with social entrepreneurial ventures such as local food projects and creative community inventions, have the potential to enable social innovation for healthy and happy communities.

Review of current test methods for screwed connections

The pull-through/local dimpling failure strength of screwed connections is very important in the design of profiled steel cladding systems to help them resist storms and hurricanes. The current American and European provisions recommend four different test methods for the screwed connections in tension, but the accuracy of these methods in determining the connection strength is not known. It is unlikely that the four test methods are equivalent in all cases and thus it is necessary to reduce the number of methods recommended. This paper presents a review of these test methods based on some laboratory tests on crest- and valley-fixed claddings and then recommends alternative tests methods that reproduce the real behavior of the connections, including the bending and membrane deformations of the cladding around the screw fasteners and the tension load in the fastener.

Local plastic mechanisms in thin steel plates under in-plane compression

Thin-walled steel plates subjected to in-plane compression develop two types of local plastic mechanism, namely the roof-shaped mechanism and the so-called flip-disc mechanism, but the intriguing question of why two mechanisms should develop was not answered until recently. It was considered that the location of first yield point shifted from the centre of the plate to the midpoint of the longitudinal edge depending on the b/t ratio, imperfection level, and yield stress of steel, which then decided the type of mechanism. This paper has verified this hypothesis using analysis and laboratory experiments. An elastic analysis using Galerkin's method to solve Marguerre's equations was first used to determine the first yield point, based on which the local plastic mechanism/imperfection tolerance tables have been developed which give the type of mechanism as a function of b/t ratio, imperfection level and yield stress of steel. Laboratory experiments of thin-walled columns verified the imperfection tolerance tables and thus indirectly the hypothesis. Elastic and rigid-plastic curves were them used to predict the effect on the ultimate load due to the change of mechanism. A finite element analysis of selected cases also confirmed the results from simple analyses and experiments.

Local buckling behaviour of steel plate elements supported by a plastic foam material

Sandwich panels comprising steel facings and a polystyrene foam core are increasingly used as roof and wall claddings in buildings in Australia. When they are subjected to loads causing bending and/or axial compression, the steel plate elements of their profiled facing are susceptible to local buckling. However, when compared to panels with no foam core, they demonstrate significantly improved local buckling behaviour because they are supported by foam. In order to quantify such improvements and to validate the use of available design buckling stress formulae, an investigation using finite element analyses and laboratory experiments was carried out on steel plates that are commonly used in Australia of varying yield stress and thickness supported by a polystyrene foam core. This paper presents the details of this investigation, the buckling results and their comparison with available design buckling formulae.

Building Healthy Communities : GP Network, Medicare Local and Primary Health Care in the Wheatbelt Region of Western Australia

• Government reports consistently recognise the importance of Primary Health Care to an efficient health system. Barriers identified in Australia’s Primary Health Care include workforce pressures, increase rate of chronic disease, and equitable access to Primary Health Care services. • General Practitioners (GPs) are the key to the successful delivery of Primary Health Care especially in rural and remote regions such as the Wheatbelt region in Western Australia (WA). • The Wheatbelt region of WA is vast: some 72,500 residents spread across 150,000km2 in 43 Local Government Authorities catchments. Majority of the Wheatbelt residents live in small towns. There is a higher reported rates of chronic disease, more at risk of chronic diseases and less utilisation of Primary Health Care services in this region. • General practice patients in the Wheatbelt are among those most in need of Primary Health Care services. • Wheatbelt GP Network (the “Network”) was established in 1998. It is a key health service delivery stakeholder in the Wheatbelt. • The Network has responded to the health needs of the community by creating a mobile Allied Health Team that works closely with GPs and is adaptive to ensure priority needs are met. • The Medicare Local model introduced by the Australian Government in 2011 aimed to improve the delivery of Primary Health Care services by improved health planning and coordinating service delivery. • Little if any recognition has been given to the outstanding work that many Divisions of General Practice have done in improving the delivery of Primary Health Care services such as the Network. • The Network has continued to support GPs and general practices and created a complementary system that integrated general practice with the work of an Allied Health Team. Its program mix is extensive. • The Network has consistently delivered on-required contract outputs and has a fifteen (15) years history of operating successfully in a large geographical area comprising in the main smaller communities that cannot support the traditional health services model. • The complexity of supporting International Medical Graduates in the region requires special attention. • The introduction of the Medicare Local in the South West of WA and their intention to take over the delivery of health services, thus effectively shutting the Network will have catastrophic consequences and cannot be supported economically. • The Network proposes to create a new model, built on its past work that increases the delivery of Primary Health Care services through its current Allied Health Team. • The proposal uses the Wheatbelt GP Super Clinic currently under construction in Northam, part of the Network and funded by the Australian Government is a key to the proposed new model. • Wheatbelt GP Super Clinic is different from existing models of GP Super Clinics around Australia which focus predominately on co-location of services. Wheatbelt GP Super Clinic utilises a hub and spoke model of service outreach to small rural towns to ensure equitable Primary Health Care coverage and continuum of care in a financially responsible and viable manner. In particular, the Wheatbelt GP Super Clinic recognises the importance of Allied Health Professionals and will involve them in a collaborative model with rural general practice. • The proposed model advocated by the Network aims to substitute the South West WA Medicare Local direct service delivery proposed for the Wheatbelt. The Network’s proposed model is to expand on the current hub and spoke model of Primary Health Care delivery to otherwise small unviable Wheatbelt towns. A flexible and adaptive skill mix of Allied Health Professionals, Nurse Practitioners and GPs ensure equitable access to service. Expanded scope of practices are utilised to reduce duplication of service and concentration of services in major towns. This involves a partnership approach. • If the proposed model not funded, the Network and the Wheatbelt region will stand to lose 16 Allied Health Professionals and defeats the purpose of Australian Government current funding for the construction of the Wheatbelt GP Super Clinic. • The Network has considered how its model can best be funded. It proposes a re-allocation of funds made available to the South West WA Medicare Local. • This submission argues that the proposal for the South West WA Medicare Local to take over the service delivery of Primary Health Care services in the Wheatbelt makes no economic sense when an existing agency (the Network) has the infrastructure in place, is experienced in working in this geographical area that has special needs and is capable to expand its programs to meet demand.

Designing for participation in local suburban communications

People have difficulty accessing means to easily publicise and discuss their ideas and concerns with their local community. The aim of this research has been to design and evaluate capacity for Internet technologies coupled with public displays to engage a diverse range of community members in the making of a shared local suburban communications network. The research problem relates to the challenges of community building, in particular discovering mechanisms that work to engage a target community and motivate participation. In an effort to understand genuine participation and barriers to use, the study was embedded in a local community and purposely longitudinal. This research contributes knowledge about the limitations of public displays to increase visibility of local communications, the need for long-term and networked visibility of community-building communications, the integral role of community facilitators, and the challenges of sustaining shared communications.

A meshfree-based local Galerkin method with condensation of degree of freedom

Condensation technique of degree of freedom is firstly proposed to improve the computational efficiency of meshfree method with Galerkin weak form. In present method, scattered nodes without connectivity are divided into several subsets by cells with arbitrary shape. The local discrete equations are established over each cell by using moving kriging interpolation, in which the nodes that located in the cell are used for approximation. Then, the condensation technique can be introduced into the local discrete equations by transferring equations of inner nodes to equations of boundary nodes based on cell. In the scheme of present method, the calculation of each cell is carried out by meshfree method with Galerkin weak form, and local search is implemented in interpolation. Numerical examples show that the present method has high computational efficiency and convergence, and good accuracy is also obtained.

New strategy toward dioxin risk reduction for local residents surrounding severe dioxin hotspots in Vietnam

Background A public health intervention program with active involvement of local related stakeholders was piloted in the Bien Hoa dioxin hot spot (2007-2009), and then expanded to the Da Nang dioxin hot spot in Vietnam (2009-2011). It aimed to reduce the risk of dioxin exposure through foods for local residents. This article presents the results of the intervention in Da Nang. Methodology To assess the results of this intervention program, pre-intervention and post-intervention knowledge-attitude-practice (KAP) surveys were implemented in 400 households, randomly selected from four wards surrounding Da Nang Airbase in 2009 and 2011, respectively. Results After the intervention, the knowledge on the existence of dioxin in food, dioxin exposure pathways, potential high risk foods and preventive measures significantly increased (p < 0.05). 98% were willing to follow advice on preventing dioxin exposure. Practices to reduce the risk of dioxin exposure also statistical significantly improved (p<0.05). After intervention, 60.4% of households undertook exposure preventive measures, significantly higher than that of the pre-intervention survey (39.6%; χ2 =40.15 , P<0.001). High risk foods had quite low rates of daily consumption (from 0% to 2.5%) and were significantly reduced (p<0.05). Conclusions This is seen as an effective intervention strategy toward reducing the risk of human exposure to dioxin at dioxin hot spots. While greater efforts are needed for remediating dioxin polluted areas inside airbases, there is also evidence to suggest that, during the past four decades, pollution has been expanding to the surrounding areas. For this reason, this model should be quickly expanded to the remaining dioxin hot spots in Vietnam to further reduce the exposure risk in these areas.

Quantitative proteomic approach to study subcellular localization of membrane proteins

As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-state distribution of hundreds of integral membrane proteins within organelles. The technique uses a partial membrane fractionation strategy in conjunction with quantitative proteomics. Localization of proteins is achieved by measuring their distribution pattern across the density gradient using amine-reactive isotope tagging and comparing these patterns with those of known organelle residents. LOPIT relies on the assumption that proteins belonging to the same organelle will co-fractionate. Multivariate statistical tools are then used to group proteins according to the similarities in their distributions, and hence localization without complete centrifugal separation is achieved. The protocol requires approximately 3 weeks to complete and can be applied in a high-throughput manner to material from many varied sources.

Sub-cellular localization of membrane proteins

In eukaryotes, numerous complex sub-cellular structures exist. The majority of these are delineated by membranes. Many proteins are trafficked to these in order to be able to carry out their correct physiological function. Assigning the sub-cellular location of a protein is of paramount importance to biologists in the elucidation of its role and in the refinement of knowledge of cellular processes by tracing certain activities to specific organelles. Membrane proteins are a key set of proteins as these form part of the boundary of the organelles and represent many important functions such as transporters, receptors, and trafficking. They are, however, some of the most challenging proteins to work with due to poor solubility, a wide concentration range within the cell and inaccessibility to many of the tools employed in proteomics studies. This review focuses on membrane proteins with particular emphasis on sub-cellular localization in terms of methodologies that can be used to determine the accurate location of membrane proteins to organelles. We also discuss what is known about the membrane protein cohorts of major organelles.

The effect of extent of local resection on patterns of disease progression in malignant pleural mesothelioma

Background We sought to determine whether or not there are differences in disease progression after radical or nonradical (debulking) surgical procedures for malignant pleural mesothelioma. Methods Over a 49-month period, 132 patients with malignant pleural mesothelioma underwent surgery. Fifty-three underwent extrapleural pneumonectomy and 79 underwent nonradical procedures. Time to evidence of clinical disease progression was recorded, as was the site(s) of that disease. Results One-hundred nineteen patients were evaluable, of which 59% (22 radical; 48 nonradical) had disease progression. Overall 30-day mortality was 8.5% (7.5% radical; 9% nonradical). The median time to overall disease progression was considerably longer after extrapleural pneumonectomy than debulking surgery (319 days vs 197 days, p = 0.019), as was the time to local disease progression (631 days vs 218 days, p = 0.0018). There was no preponderance of earlier stage disease in the radical surgery group. There was a trend toward prolonged survival in those undergoing radical surgery, but no significant difference between the groups (497 days vs 324 days, p = 0.079). In those who had extrapleural pneumonectomy, time-to-disease progression significantly decreased with N2 disease compared with N0/1 involvement (197 days vs 358 days, p = 0.02). Conclusions Extrapleural pneumonectomy may be preferable to debulking surgery in malignant pleural mesothelioma to delay disease progression and give greater control of local disease. Involvement of N2 nodes is associated with accelerated disease progression and is therefore a contraindication to extrapleural pneumonectomy. © 2004 by The Society of Thoracic Surgeons.

Advanced Engineering of Lipid Metabolism in Nicotiana benthamiana Using a Draft Genome and the V2 Viral Silencing-Suppressor Protein

The transient leaf assay in Nicotiana benthamiana is widely used in plant sciences, with one application being the rapid assembly of complex multigene pathways that produce new fatty acid profiles. This rapid and facile assay would be further improved if it were possible to simultaneously overexpress transgenes while accurately silencing endogenes. Here, we report a draft genome resource for N. benthamiana spanning over 75% of the 3.1 Gb haploid genome. This resource revealed a two-member NbFAD2 family, NbFAD2.1 and NbFAD2.2, and quantitative RT-PCR (qRT-PCR) confirmed their expression in leaves. FAD2 activities were silenced using hairpin RNAi as monitored by qRT-PCR and biochemical assays. Silencing of endogenous FAD2 activities was combined with overexpression of transgenes via the use of the alternative viral silencing-suppressor protein, V2, from Tomato yellow leaf curl virus. We show that V2 permits maximal overexpression of transgenes but, crucially, also allows hairpin RNAi to operate unimpeded. To illustrate the efficacy of the V2-based leaf assay system, endogenous lipids were shunted from the desaturation of 18:1 to elongation reactions beginning with 18:1 as substrate. These V2-based leaf assays produced ~50% more elongated fatty acid products than p19-based assays. Analyses of small RNA populations generated from hairpin RNAi against NbFAD2 confirm that the siRNA population is dominated by 21 and 22 nt species derived from the hairpin. Collectively, these new tools expand the range of uses and possibilities for metabolic engineering in transient leaf assays. © 2012 Naim et al.

The Enamovirus P0 protein is a silencing suppressor which inhibits local and systemic RNA silencing through AGO1 degradation

The P0 protein of poleroviruses and P1 protein of sobemoviruses suppress the plant's RNA silencing machinery. Here we identified a silencing suppressor protein (SSP), P0PE, in the Enamovirus Pea enation mosaic virus-1 (PEMV-1) and showed that it and the P0s of poleroviruses Potato leaf roll virus and Cereal yellow dwarf virus have strong local and systemic SSP activity, while the P1 of Sobemovirus Southern bean mosaic virus supresses systemic silencing. The nuclear localized P0PE has no discernable sequence conservation with known SSPs, but proved to be a strong suppressor of local silencing and a moderate suppressor of systemic silencing. Like the P0s from poleroviruses, P0PE destabilizes AGO1 and this action is mediated by an F-box-like domain. Therefore, despite the lack of any sequence similarity, the poleroviral and enamoviral SSPs have a conserved mode of action upon the RNA silencing machinery. © 2012 Elsevier Inc.