Eimeria tenella protein trafficking: differential regulation of secretion versus surface tethering during the life cycle

Eimeria spp. are intracellular parasites that have a major impact on poultry. Effective live vaccines are available and the development of reverse genetic technologies has raised the prospect of using Eimeria spp. as recombinant vectors to express additional immunoprotective antigens. To study the ability of Eimeria to secrete foreign antigens or display them on the surface of the sporozoite, transiently transfected populations of E. tenella expressing the fluorescent protein mCherry, linked to endogenous signal peptide (SP) and glycophosphatidylinositol-anchor (GPI) sequences, were examined. The SP from microneme protein EtMIC2 (SP2) allowed efficient trafficking of mCherry to cytoplasmic vesicles and following the C-terminal addition of a GPI-anchor (from surface antigen EtSAG1) mCherry was expressed on the sporozoite surface. In stable transgenic populations, mCherry fused to SP2 was secreted into the sporocyst cavity of the oocysts and after excystation, secretion was detected in culture supernatants but not into the parasitophorous vacuole after invasion. When the GPI was incorporated, mCherry was observed on the sporozites surface and in the supernatant of invading sporozoites. The proven secretion and surface exposure of mCherry suggests that antigen fusions with SP2 and GPI of EtSAG1 may be promising candidates to examine induction of protective immunity against heterologous pathogens

Viral proteins expressed in the protozoan parasite Eimeria tenella are detected by the chicken immune system

BACKGROUND: Eimeria species are parasitic protozoa that cause coccidiosis, an intestinal disease commonly characterised by malabsorption, diarrhoea and haemorrhage that is particularly important in chickens. Vaccination against chicken coccidiosis is effective using wild-type or attenuated live parasite lines. The development of protocols to express foreign proteins in Eimeria species has opened up the possibility of using Eimeria live vaccines to deliver heterologous antigens and function as multivalent vaccine vectors that could protect chickens against a range of pathogens. RESULTS: In this study, genetic complementation was used to express immunoprotective virus antigens in Eimeria tenella. Infectious bursal disease virus (IBDV) causes Gumboro, an immunosuppressive disease that affects productivity and can interfere with the efficacy of poultry vaccination programmes. Infectious laryngotracheitis virus (ILTV) causes a highly transmissible respiratory disease for which strong cellular immunity and antibody responses are required for effective vaccination. Genes encoding the VP2 protein from a very virulent strain of IBDV (vvVP2) and glycoprotein I from ILTV (gI) were cloned downstream of 5’Et-Actin or 5’Et-TIF promoter regions in plasmids that also contained a mCitrine fluorescent reporter cassette under control of the 5’Et-MIC1 promoter. The plasmids were introduced by nucleofection into E. tenella sporozoites, which were then used to infect chickens. Progeny oocysts were sorted by FACS and passaged several times in vivo until the proportion of fluorescent parasites in each transgenic population reached ~20 % and the number of transgene copies per parasite genome decreased to < 10. All populations were found to transcribe and express the transgene and induced the generation of low titre, transgene-specific antibodies when used to immunise chickens. CONCLUSIONS: E. tenella can express antigens of other poultry pathogens that are successfully recognised by the chicken immune system. Nonetheless, further work has to be done in order to improve the levels of expression for its future use as a multivalent vaccine vector. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-016-1756-2) contains supplementary material, which is available to authorized users

The Price of Slavery: An Analysis of Human Trafficking Policy and Spending in Department of Defense Procurement

Excerpt from the Proceedings of the Nineteenth Annual Acquisition Research SymposiumThe Department of Defense (DoD) is charged with upholding the U.S. zero tolerance human trafficking policy in a world of complex, opaque supply chains and constrained human and capital resources. This study explores how the DoD can better leverage its acquisition workforce, sourcing expertise, and data to rigorously uphold the U.S. policy of zero tolerance for human trafficking in such an environment. As part of this analysis a program evaluation of the DoD’s Combating Trafficking in Persons training was conducted for the acquisition workforce. This evaluation categorized the training related to prevention, monitoring, and response to human trafficking. A quantitative analysis of DoD spending was conducted to illuminate the amount of tax dollars spent in categories, based on product and service codes, where trafficking is most prevalent to help the DoD focus its efforts for program improvement. The spend analysis revealed areas that are most at risk. Globally, an estimated 24.9 million people are subjected to human trafficking, which generates an estimated $150 billion annually in illicit profits (White House, 2020, p. 8). Specifically, the DoD spent$13.1 billion (2018–2020) in countries that are not making significant efforts to combat human trafficking according to the Department of State. The research provides insight and recommendations on where the DoD should focus attention to address human trafficking in contracts constrained of prevention, monitoring, and response resources. Finally, we provided recommended courses of action to increase participation and enhance the mandatory training for the acquisition workforce.Approved for public release; distribution is unlimited

THE PRICE OF SLAVERY: AN ANALYSIS OF HUMAN TRAFFICKING POLICY AND SPENDING IN DEPARTMENT OF DEFENSE PROCUREMENT

The Department of Defense (DOD) is charged with upholding the U.S. zero-tolerance human trafficking policy in a world of complex, opaque supply chains and constrained human and capital resources. This study explores how the DOD can better leverage its acquisition workforce, sourcing expertise, and data to rigorously uphold U.S. policy of zero-tolerance for human trafficking in such an environment. As part of this analysis a program evaluation of the DOD’s Combating Trafficking in Persons training was conducted for the acquisition workforce. This evaluation categorized the training related to prevention, monitoring, and response to human trafficking. A quantitative analysis of DOD spend was conducted to illuminate the amount of tax dollars spent in categories, based on product and service codes, where trafficking is most prevalent to help the DOD focus its efforts for program improvement. The spend analysis revealed areas that are most at risk. Specifically, the DOD spent $13.1 billion (2018–2020) in countries that are not making significant efforts to combat human trafficking according to the Department of State. The research provides insight and recommendations on where the DOD should focus attention to address human trafficking in contracts constrained of prevention, monitoring, and response resources. Finally, we provided recommended courses of action to increase participation and enhance the mandatory training for the acquisition workforce.Outstanding ThesisFirst Lieutenant, United States Air ForceCaptain, United States Air ForceFirst Lieutenant, United States Air ForceApproved for public release. Distribution is unlimited

Meta-analysis of the Sea Change Consultation Reports

Sea Change is a three-year, EU funded, Horizon 2020 project with 17 partners in nine European countries (Belgium, Denmark, Greece, Sweden, France, Ireland, Portugal, Spain and UK). During April and May 2016, eight of these countries (except France) carried out consultations with education stakeholders (classified as incumbents, regulating agencies and challengers, see p. xii) for the purpose of gaining deeper insights into the barriers to teaching 12-19 year olds about the ocean, and to identify options for overcoming these barriers. The Sea Change partners used the method Collective intelligence (CI) to consult with the education stakeholders. The CI method used facilitation and problem solving to harmonise input from education stakeholders from different backgrounds and perspectives. This report was built on the Collective Intelligence consultation work completed and reported in Sea For Society (D2.4 Global Analysis of the SFS Consultation Process from a Social Sciences Perspective), as reported in Domegan et al., 2014

Implementation, impact and costs of policies for safe staffing in acute NHS trusts

The aim of this research has been to describe the implementation of safe staffing policies in NHS general acute Trusts in England looking at costs and consequences, and examining the factors that have influenced implementation. A mix of qualitative and quantitative methods were used (national survey, analysis of national data, four case studies, realist evaluation) to examine the impact of policies nationally, and explore commonality and variation in local responses to safe staffing policies. The report is based on independent research commissioned and funded by the NIHR Policy Research Programme (“Implementation, Impact and Costs of Policies for Safe Staffing in Acute NHS Trusts”, PR-ST-1115-10017). The views expressed are those of the authors and not necessarily those of the NIHR, the Department of Health and Social Care, ‘arm’s length bodies’ or other government departments

The New Political Economy of EU State Aid Policy

Despite its importance and singularity, the EU’s state aid policy has attracted less scholarly attention than other elements of EU competition policy. Introducing the themes addressed by the special issue, this article briefly reviews the development of EU policy and highlights why the control of state aid matters. The Commission’s response to the current economic crisis notably in banking and the car industry is a key concern, but the interests of the special issue go far beyond. They include: the role of the European Commission in the development of EU policy, the politics of state aid, and a clash between models of capitalism. The special issue also examines the impact of EU policy. It investigates how EU state aid decisions affect not only industrial policy at the national level (and therefore at the EU level), but the welfare state and territorial relations within federal member states, the external implications of EU action and the strategies pursued by the Commission to limit any potential disadvantage to European firms, and the conflict between the EU’s expanding legal order and national

Three Dimensional MHD Wave Propagation and Conversion to Alfven Waves near the Solar Surface. I. Direct Numerical Solution

The efficacy of fast/slow MHD mode conversion in the surface layers of sunspots has been demonstrated over recent years using a number of modelling techniques, including ray theory, perturbation theory, differential eigensystem analysis, and direct numerical simulation. These show that significant energy may be transferred between the fast and slow modes in the neighbourhood of the equipartition layer where the Alfven and sound speeds coincide. However, most of the models so far have been two dimensional. In three dimensions the Alfven wave may couple to the magneto-acoustic waves with important implications for energy loss from helioseismic modes and for oscillations in the atmosphere above the spot. In this paper, we carry out a numerical ``scattering experiment'', placing an acoustic driver 4 Mm below the solar surface and monitoring the acoustic and Alfvenic wave energy flux high in an isothermal atmosphere placed above it. These calculations indeed show that energy conversion to upward travelling Alfven waves can be substantial, in many cases exceeding loss to slow (acoustic) waves. Typically, at penumbral magnetic field strengths, the strongest Alfven fluxes are produced when the field is inclined 30-40 degrees from the vertical, with the vertical plane of wave propagation offset from the vertical plane containing field lines by some 60-80 degrees.Comment: Accepted for the HELAS II/ SOHO 19/ GONG 2007 Topical Issue of Solar Physic

NEXUS/Physics: An interdisciplinary repurposing of physics for biologists

In response to increasing calls for the reform of the undergraduate science curriculum for life science majors and pre-medical students (Bio2010, Scientific Foundations for Future Physicians, Vision & Change), an interdisciplinary team has created NEXUS/Physics: a repurposing of an introductory physics curriculum for the life sciences. The curriculum interacts strongly and supportively with introductory biology and chemistry courses taken by life sciences students, with the goal of helping students build general, multi-discipline scientific competencies. In order to do this, our two-semester NEXUS/Physics course sequence is positioned as a second year course so students will have had some exposure to basic concepts in biology and chemistry. NEXUS/Physics stresses interdisciplinary examples and the content differs markedly from traditional introductory physics to facilitate this. It extends the discussion of energy to include interatomic potentials and chemical reactions, the discussion of thermodynamics to include enthalpy and Gibbs free energy, and includes a serious discussion of random vs. coherent motion including diffusion. The development of instructional materials is coordinated with careful education research. Both the new content and the results of the research are described in a series of papers for which this paper serves as an overview and context.Comment: 12 page

On the making and taking of professionalism in the further education workplace

This paper examines the changing nature of professional practice in English further education. At a time when neo-liberal reform has significantly impacted on this under-researched and over-market-tested sector, little is known about who its practitioners are and how they construct meaning in their work. Sociological interest in the field has tended to focus on further education practitioners as either the subjects of market and managerial reform or as creative agents operating within the contradictions of audit and inspection cultures. In challenging such dualism, which is reflective of wider sociological thinking, the paper examines the ways in which agency and structure combine to produce a more transformative conception of the further education professional. The approach contrasts with a prevailing policy discourse that seeks to re-professionalise and modernise further education practice without interrogating either the terms of its professionalism or the neo-liberal practices in which it resides