In the enteric pathogen Salmonella enterica serovar Typhimurium, invasion and motility are coordinated by the master regulator HilD, which induces expression of the type III secretion system 1 (T3SS1) and motility genes. Methyl-accepting chemotaxis proteins (MCPs) detect specific ligands and control the direction of the flagellar motor, promoting tumbling and changes in direction (if a repellent is detected) or smooth swimming (in the presence of an attractant). Here, we show that HilD induces smooth swimming by upregulating an uncharacterized MCP (McpC), and this is important for invasion of epithelial cells. Remarkably, in vitro assays show that McpC can suppress tumbling and increase smooth swimming in the absence of exogenous ligands. Expression of mcpC is repressed by the universal regulator H-NS, which can be displaced by HilD. Our results highlight the importance of smooth swimming for Salmonella Typhimurium invasiveness and indicate that McpC can act via a ligand-independent mechanism when incorporated into the chemotactic receptor array.

Classical Swine Fever Virus (CSFV) causes classical swine fever, a highly contagious hemorrhagic fever affecting both feral and domesticated pigs. Outbreaks of CSF in Europe, Asia, Africa and South America had significant adverse impacts on animal health, food security and the pig industry. The disease is generally contained by prevention of exposure through import restrictions (e.g. banning import of live pigs and pork products), localized vaccination programmes and culling of infected or at-risk animals, often at very high cost. Current CSFV-modified live virus vaccines are protective, but do not allow differentiation of infected from vaccinated animals (DIVA), a critical aspect of disease surveillance programmes. Alternatively, first-generation subunit vaccines using the viral protein E2 allow for use of DIVA diagnostic tests, but are slow to induce a protective response, provide limited prevention of vertical transmission and may fail to block viral shedding. CSFV E2 subunit vaccines from a baculovirus/insect cell system have been developed for several vaccination campaigns in Europe and Asia. However, this expression system is considered expensive for a veterinary vaccine and is not ideal for wide-spread deployment. To address the issues of scalability, cost of production and immunogenicity, we have employed an Agrobacterium-mediated transient expression platform in Nicotiana benthamiana and formulated the purified antigen in novel oil-in-water emulsion adjuvants. We report the manufacturing of adjuvanted, plant-made CSFV E2 subunit vaccine. The vaccine provided complete protection in challenged pigs, even after single-dose vaccination, which was accompanied by strong virus neutralization antibody responses.

A reverse vaccinology system, Vaxign, was used to identify and select a subset of five African Swine Fever (ASF) antigens that were successfully purified from human embryonic kidney 293 (HEK) cells and produced in Modified vaccinia virus Ankara (MVA) viral vectors. Three HEK-purified antigens [B646L (p72), E183L (p54), and O61R (p12)], and three MVA-vectored antigens [B646L, EP153R, and EP402R (CD2v)] were evaluated using a prime-boost immunization regimen swine safety and immunogenicity study. Antibody responses were detected in pigs following prime-boost immunization four weeks apart with the HEK-293-purified p72, p54, and p12 antigens. Notably, sera from the vaccinees were positive by immunofluorescence on ASFV (Georgia 2007/1)-infected primary macrophages. Although MVA-vectored p72, CD2v, and EP153R failed to induce antibody responses, interferon-gamma (IFN-γ+) spot forming cell responses against all three antigens were detected one week post-boost. The highest IFN-γ+ spot forming cell responses were detected against p72 in pigs primed with MVA-p72 and boosted with the recombinant p72. Antigen-specific (p12, p72, CD2v, and EP153R) T-cell proliferative responses were also detected post-boost. Collectively, these results are the first demonstration that ASFV subunit antigens purified from mammalian cells or expressed in MVA vectors are safe and can induce ASFV-specific antibody and T-cell responses following a prime-boost immunization regimen in swine.

Characterization of host microbial interactions typically occurs on the cellular or protein level. Recently, a more thorough and accurate appreciation of cellular interactions has come into better focus with improved understanding of membrane vesicles (OMV). While OMVs are documented primarily in Gram-negative bacteria, certain Gram-positive species generate these structures, despite the obvious physical limitations of the cell envelope. Here, we briefly review the current understanding of OMVs in content and function, their role in pathogenesis, and the consequences of somatic cell gene expression on these events.

Rift Valley fever Virus (RVFV), a negative-stranded RNA virus, is the etiological agent of the vector-borne zoonotic disease, Rift Valley fever (RVF). In both humans and livestock, protective immunity can be achieved through vaccination. Earlier and more recent vaccine trials in cattle and sheep demonstrated a strong neutralizing antibody and total IgG response induced by the RVF vaccine, authentic recombinant MP-12 (arMP-12). From previous work, protective immunity in sheep and cattle vaccinates normally occurs from 7 to 21 days after inoculation with arMP-12. While the serology and protective response induced by arMP-12 has been studied, little attention has been paid to the underlying molecular and genetic events occurring prior to the serologic immune response. To address this, we isolated RNA from whole blood of vaccinated calves over a time course of 21 days before and after vaccination with arMP-12. The time course RNAs were sequenced by RNASeq and bioinformatically analyzed. Our results revealed time-dependent activation or repression of numerous gene ontologies and pathways related to the vaccine induced immune response and its regulation. Additional bioinformatic analyses identified a correlative relationship between specific host immune response genes and protective immunity prior to the detection of protective serum neutralizing antibody responses. These results contribute an important proof of concept for identifying molecular and genetic components underlying the immune response to RVF vaccination and protection prior to serologic detection.

The purpose of this paper is to reflect on the life of Tony Lowe, Emeritus Professor of Accounting and Financial Management at the University of Sheffield, who died on 5 March 2014. It celebrates Tony Lowe’s considerable direct contributions to accounting knowledge and, possibly more significantly, his indirect contribution through his enabling of a range of those associated with him at Sheffield to become scholars of distinction in their own right. Apart from providing insight into Tony Lowe’s direct contribution to accounting knowledge through an analysis of a range of significant sole authored and joint authored publications, the paper gives rather more attention to his more indirect enabling contribution. In this regard, it traces the development of initially the Management Control Association and subsequently the “Sheffield School” to Tony Lowe, clarifying the values that underlie these groups. It also clarifies how some of the key elements that have allowed the now global Interdisciplinary and Critical Perspectives on Accounting (ICPA) Project to exist and flourish are traceable to Tony Lowe and the “Sheffield School” he created.

Abstract This article summarises the extensive management changes, broadly and generically referred to as the New Public Management (NPM), or, the New Public Financial Management (NPFM) reforms as they are more accurately described, that have been occurring in the public services in the United Kingdom and other developed countries across the world. It demonstrates not only the diversity and increasing intensity of these NPFM changes but also the centrality of accounting, in its many different forms, to not only enable these changes but also to define and shape their nature. The article also demonstrates the power and significance of a range of qualitative and interpretive accounting research, which goes beyond seeing accounting as little more than a technology, to provide contextually‐rich understandings of the central role of accounting in defining and enabling these NPFM changes.

Membrane vesicles (MVs), discrete nano-structures produced from the outer membrane of Gram-negative bacteria such as Salmonella enterica Typhimurium (S. Typhimurium), strongly activate dendritic cells (DCs), contain major antigens (Ags) recognized by Salmonella-specific B-cells and CD4+ T-cells, and provide protection against S. Typhimurium challenge in a mouse model. With this in mind, we hypothesized that alterations to the gene expression profile of bacteria will be reflected in the immunologic response to MVs. To test this, we assessed the ability of MVs from wild-type (WT) S. Typhimurium or a strain with a phenotype mimicking the intracellular-phase of S. Typhimurium (PhoPc) to activate dendritic cells and initiate a strong inflammatory response. MVs, isolated from wild-type and PhoPc S. Typhimurium (WTMVs and PhoPcMVs, respectively) had pro-inflammatory properties consistent with the parental bacterial strains: PhoPcMVs were less stimulatory for DC activation in vitro and were impaired for subsequent inflammatory responses compared to WTMVs. Interestingly, the reduced pro-inflammatory properties of PhoPcMVs did not completely rely on signals through TLR4, the receptor for LPS. Nonetheless, both WTMVs and PhoPcMVs contained abundant immunogenic antigens capable of being recognized by memory-immune CD4+ T-cells from mice previously infected with S. Typhimurium. Furthermore, we analyzed a suite of pathogenic Gram-negative bacteria and their purified MVs for their ability to activate DCs and stimulate inflammation in a manner consistent with the known inflammatory properties of the parental strains, as shown for S. Typhimurium. Finally, analysis of the potential vaccine utility of S. Typhimurium MVs revealed their capacity to encapsulate an exogenous model antigen and stimulate antigen-specific CD4+ and CD8+ T-cell responses. Taken together, our results demonstrate the dependence of bacterial cell gene expression for MV immunogenicity and subsequent in vitro immunologic response, as well as their potential utility as a vaccine platform.

Purpose - The purpose of this paper is to reflect on the life of Tony Lowe, Emeritus Professor of Accounting and Financial Management at the University of Sheffield, who died on 5 March 2014. It celebrates Tony Lowe's considerable direct contributions to accounting knowledge and, possibly more significantly, his indirect contribution through his enabling of a range of those associated with him at Sheffield to become scholars of distinction in their own right. Design/methodology/approach - Publication review, personal reflections and argument. Findings - Apart from providing insight into Tony Lowe's direct contribution to accounting knowledge through an analysis of a range of significant sole authored and joint authored publications, the paper gives rather more attention to his more indirect enabling contribution. In this regard it traces the development of initially the Management Control Association and subsequently the "Sheffield School" to Tony Lowe, clarifying the values that underlie these groups. It also clarifies how some of the key elements that have allowed the now global Interdisciplinary and Critical Perspectives on Accounting (ICPA) Project to exist and flourish are traceable to Tony Lowe and the "Sheffield School" he created. Research limitations/implications - This paper provides an important historical analysis of the direct and indirect influence of a unique scholar on the beginnings and development of particularly the now global ICPA Project. This history is personal and maybe selective and possibly limited because of this but hopefully will encourage others to investigate the claims further. Originality/value - The history of the ICPA Project has only partially been told before. This is another part of this history that has not been analysed before on which further work can build.

This paper comes from comments made at an event at the 2013 CSEAR Summer School to celebrate the considerable and ongoing contributions of Professor Rob Gray to the development of Social and Environmental Accounting (SEA). It concentrates on Rob Gray's contributions to the pressing agenda for far-reaching organisational change to reverse the considerable range of social and environmental problems that exist in our societies, to the nature and role of SEA in this change agenda and in changing the nature of financial reporting and regulation more generally. The paper also explores some of Rob Gray's ongoing concerns that this much needed organisational change is partial at best and that SEA, as currently articulated, is not providing the vehicle for enabling the levels of change that are required.

, 708 (2013); 339 Science et al. Sebastian E. Winter in the Inflamed Gut E. coli Host-Derived Nitrate Boosts Growth of This copy is for your personal, non-commercial use only. clicking here. colleagues, clients, or customers by , you can order high-quality copies for your If you wish to distribute this article to others here. following the guidelines can be obtained by Permission to republish or repurpose articles or portions of articles ): July 8, 2014 www.sciencemag.org (this information is current as of The following resources related to this article are available online at http://www.sciencemag.org/content/339/6120/708.full.html version of this article at: including high-resolution figures, can be found in the online Updated information and services, http://www.sciencemag.org/content/suppl/2013/02/07/339.6120.708.DC1.html can be found at: Supporting Online Material http://www.sciencemag.org/content/339/6120/708.full.html#related found at: can be related to this article A list of selected additional articles on the Science Web sites http://www.sciencemag.org/content/339/6120/708.full.html#ref-list-1 , 20 of which can be accessed free: cites 36 articles This article http://www.sciencemag.org/content/339/6120/708.full.html#related-urls 12 articles hosted by HighWire Press; see: cited by This article has been http://www.sciencemag.org/cgi/collection/microbio Microbiology subject collections: This article appears in the following

Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying Salmonella-host interactions are important to elucidate the molecular pathogenesis of Salmonella infection. The Gentamicin protection assay to phenotype Salmonella association, invasion and replication in phagocytic cells was adapted to allow high-throughput screening to define the roles of deletion mutants of Salmonella enterica serotype Typhimurium in host interactions using RAW 264.7 murine macrophages. Under this protocol, the variance in measurements is significantly reduced compared to the standard protocol, because wild-type and multiple mutant strains can be tested in the same culture dish and at the same time. The use of multichannel pipettes increases the throughput and enhances precision. Furthermore, concerns related to using less host cells per well in 96-well culture dish were addressed. Here, the protocol of the modified in vitro Salmonella invasion assay using phagocytic cells was successfully employed to phenotype 38 individual Salmonella deletion mutants for association, invasion and intracellular replication. The in vitro phenotypes are presented, some of which were subsequently confirmed to have in vivo phenotypes in an animal model. Thus, the modified, standardized assay to phenotype Salmonella association, invasion and replication in macrophages with high-throughput capacity could be utilized more broadly to study bacterial-host interactions.

For over 25 years, we have undertaken research that has used a research approach called Middle-Range Thinking (MRT) (cf. Laughlin, 1995, 2004; Broadbent & Laughlin, 2008) as a means of seeking to develop understanding of the role of accounting in change in the social world. In considering change in both society and organizations, we have concentrated on wide-ranging management controls, particularly those using accounting technologies, which have been implemented in the attempt to implement the required change. This chapter explores some elements of MRT both generally and specifically in the context of understanding control in conditions of uncertainty.

ABSTRACT To establish a replicative niche during its infectious cycle between the intestinal lumen and tissue, the enteric pathogen Salmonella enterica serovar Typhimurium requires numerous virulence genes, including genes for two type III secretion systems (T3SS) and their cognate effectors. To better understand the host-pathogen relationship, including early infection dynamics and induction kinetics of the bacterial virulence program in the context of a natural host, we monitored the subcellular localization and temporal expression of T3SS-1 and T3SS-2 using fluorescent single-cell reporters in a bovine, ligated ileal loop model of infection. We observed that the majority of bacteria at 2 h postinfection are flagellated, express T3SS-1 but not T3SS-2, and are associated with the epithelium or with extruding enterocytes. In epithelial cells, S. Typhimurium cells were surrounded by intact vacuolar membranes or present within membrane-compromised vacuoles that typically contained numerous vesicular structures. By 8 h postinfection, T3SS-2-expressing bacteria were detected in the lamina propria and in the underlying mucosa, while T3SS-1-expressing bacteria were in the lumen. Our work identifies for the first time the temporal and spatial regulation of T3SS-1 and -2 expression during an enteric infection in a natural host and provides further support for the concept of cytosolic S. Typhimurium in extruding epithelium as a mechanism for reseeding the lumen. IMPORTANCE The pathogenic bacterium Salmonella enterica serovar Typhimurium invades and persists within host cells using distinct sets of virulence genes. Genes from Salmonella pathogenicity island 1 (SPI-1) are used to initiate contact and facilitate uptake into nonphagocytic host cells, while genes within SPI-2 allow the pathogen to colonize host cells. While many studies have identified bacterial virulence determinants in animal models of infection, very few have focused on virulence gene expression at the single-cell level during an in vivo infection. To better understand when and where bacterial virulence factors are expressed during an acute enteric infection of a natural host, we infected bovine jejunal-ileal loops with S. Typhimurium cells harboring fluorescent transcriptional reporters for SPI-1 and -2 (PinvF and PssaG, respectively). After a prescribed time of infection, tissue and luminal fluid were collected and analyzed by microscopy. During early infection (≤2 h), bacteria within both intact and compromised membrane-bound vacuoles were observed within the epithelium, with the majority expressing SPI-1. As the infection progressed, S. Typhimurium displayed differential expression of the SPI-1 and SPI-2 regulons, with the majority of tissue-associated bacteria expressing SPI-2 and the majority of lumen-associated bacteria expressing SPI-1. This underscores the finding that Salmonella virulence gene expression changes as the pathogen transitions from one anatomical location to the next.