Expression and crystallization of SeDsbA, SeDsbL and SeSrgA from Salmonella enterica serovar Typhimurium

Pathogens require protein-folding enzymes to produce functional virulence determinants. These foldases include the Dsb family of proteins, which catalyze oxidative folding in bacteria. Bacterial disulfide catalytic processes have been well characterized in Escherichia coli K-12 and these mechanisms have been extrapolated to other organisms. However, recent research indicates that the K-12 complement of Dsb proteins is not common to all bacteria. Importantly, many pathogenic bacteria have an extended arsenal of Dsb catalysts that is linked to their virulence. To help to elucidate the process of oxidative folding in pathogens containing a wide repertoire of Dsb proteins, Salmonella enterica serovar Typhimurium has been focused on. This Gram-negative bacterium contains three DsbA proteins: SeDsbA, SeDsbL and SeSrgA. Here, the expression, purification, crystallization and preliminary diffraction analysis of these three proteins are reported. SeDsbA, SeDsbL and SeSrgA crystals diffracted to resolution limits of 1.55, 1.57 and 2.6 Å and belonged to space groups P21, P21212 and C2, respectively.

Escherichia coli isolates causing asymptomatic bacteriuria in catheterized and noncatheterized individuals possess similar virulence properties

Urinary tract infections (UTIs) are among the most common infectious diseases of humans, with Escherichia coli being responsible for >80% of all cases. Asymptomatic bacteriuria (ABU) occurs when bacteria colonize the urinary tract without causing clinical symptoms and can affect both catheterized patients (catheter-associated ABU [CA-ABU]) and noncatheterized patients. Here, we compared the virulence properties of a collection of ABU and CA-ABU nosocomial E. coli isolates in terms of antibiotic resistance, phylogenetic grouping, specific UTI-associated virulence genes, hemagglutination characteristics, and biofilm formation. CA-ABU isolates were similar to ABU isolates with regard to the majority of these characteristics; exceptions were that CA-ABU isolates had a higher prevalence of the polysaccharide capsule marker genes kpsMT II and kpsMT K1, while more ABU strains were capable of mannose-resistant hemagglutination. To examine biofilm growth in detail, we performed a global gene expression analysis with two CA-ABU strains that formed a strong biofilm and that possessed a limited adhesin repertoire. The gene expression profile of the CA-ABU strains during biofilm growth showed considerable overlap with that previously described for the prototype ABU E. coli strain, 83972. This is the first global gene expression analysis of E. coli CA-ABU strains. Overall, our data suggest that nosocomial ABU and CA-ABU E. coli isolates possess similar virulence profiles.

Regulatory interplay between pap operons in uropathogenic Escherichia coli

Pathogenic bacteria have a large repertoire of surface organelles involved in adherence, motility and protein export, but how individual bacteria co-ordinate surface organelle expression to prevent interference and excessive immune stimulation is unclear. Phase variation is a mechanism by which expression of surface factors is limited to a fraction of the bacterial population; however, the presence of multiple homologous surface structures controlled by related mechanisms and regulators antagonizes the independent expression achieved by phase variation. To investigate whether other mechanisms have evolved to sort out the bacterial cell surface, we examined regulatory cross-talk between multiple phase-variable pyelonephritis-associated pili (pap) operons in Escherichia coli isolates associated with urinary tract infections. Allelic variation identified in the regulatory regions and regulators acts synergistically to limit coexpression of homologous fimbrial operons. In particular, there is evidence that papI is under positive selection and PapI variants displayed differences in their capacity to activate related pap operons. Alleles of the high-affinity binding site for PapB were shown to contain a variable number of (T/A)3 repeats occurring every 9 bp that altered the sensitivity of pap operon activation. Taken together with other examples of surface organelle cross-talk, we illustrate how this regulation could promote sequential expression.

Turning event logs into process movies: animating what has really happened

Today’s information systems log vast amounts of data. These collections of data (implicitly) describe events (e.g. placing an order or taking a blood test) and, hence, provide information on the actual execution of business processes. The analysis of such data provides an excellent starting point for business process improvement. This is the realm of process mining, an area which has provided a repertoire of many analysis techniques. Despite the impressive capabilities of existing process mining algorithms, dealing with the abundance of data recorded by contemporary systems and devices remains a challenge. Of particular importance is the capability to guide the meaningful interpretation of “oceans of data” by process analysts. To this end, insights from the field of visual analytics can be leveraged. This article proposes an approach where process states are reconstructed from event logs and visualised in succession, leading to an animated history of a process. This approach is customisable in how a process state, partially defined through a collection of activity instances, is visualised: one can select a map and specify a projection of events on this map based on the properties of the events. This paper describes a comprehensive implementation of the proposal. It was realised using the open-source process mining framework ProM. Moreover, this paper also reports on an evaluation of the approach conducted with Suncorp, one of Australia’s largest insurance companies.

Mentors’ and mentees’ views about desirable attributes and practices for preservice teachers

Universities provide preservice teachers (mentees) with directions on how to engage within their school placements, yet mentees have ongoing concerns about how to interact with their mentors. What are desirable attributes and practices for preservice teachers in their roles as mentees? This qualitative study gathers data from primary and secondary Australian teachers (n=25) through extended response questionnaire and audio-recorded focus group discussions, and preservice teachers (n=10) using audio-recorded interviews at the conclusion of their four-week practicum. Findings indicated that mentors had clear views on desirable attributes (e.g., enthusiasm, commitment, resilience) and practices (e.g., planning, preparation, building a teaching repertoire) for mentees; whereas mentees had varying views on identifying such attributes and practices and did not refer to: content knowledge, differentiation, and knowing school and university policies. This showed that mentees need more guidance for focusing on specific attributes and practices to benefit the mentoring relationship and their teaching development.

Mentor feedback : models, viewpoints and strategies

A mentor’s feedback can present professional insights to allow a mentee to reflect and develop practice. This paper positions two models for feedback that have emanated from empirical studies. It also demonstrates the diverse viewpoints of mentors and suggests strategies for providing quality feedback. In one qualitative study, 24 mentors observed a final-year preservice teacher through a professionally video-recorded lesson and wrote their observations towards giving feedback to the potential mentee. Tables illustrated in the paper, show that mentors’ positive feedback and constructive criticisms vary considerably on the same observed events. Data from this study were synthesised to posit a theoretical model for analysing mentor feedback in an interconnected, three-way Venn diagram, namely: visual, auditory and conceptual frames. Another study (n=28), which is a collection of mentor teachers’ work samples during the Mentoring for Effective Teaching (MET) program, provides strategies within six feedback practices, that is: (1) negotiated mentor-mentee expectations for providing feedback on practices, (2) reviewing teaching plans, (3) arranging for observations of practices, (4) providing oral feedback, (5) providing written feedback, and; (6) presenting opportunities for the mentee to evaluate teaching practices with consideration of the mentor’s feedback. For example, on the last mentioned practice (6) there were strategies such as “Plan a time for evaluation of practices (guided reflection)”, “Read the mentee’s reflection on practice and discuss how it aligns with your observations of their practices”, and “Highlight verbally and/or in writing where the mentee is perceptive about the reflection and how the reflection could be enhanced for future evaluations”. Developing a range of strategies that may assist the mentee in professional growth, include enlisting a community of mentors, ensuring mentors have a repertoire of strategies for articulating feedback, and using mentor feedback tools and models. This study has implications for the development of feedback models and strategies.

Curriculum reform in testing and accountability contexts

Recent international educational developments have important implications for the skills and understandings in curriculum and assessment that teachers develop, both in pre-service and in practice. Global developments in curriculum and assessment reform require teachers to utilise a network of knowledges and develop a repertoire of assessment skills and understandings. In a context of testing, accountability and auditing, data analysis skills are increasingly required to examine pedagogic practices for the development of intervention teaching and learning strategies to improve learning outcomes for all students (Marsh, 2009). However, too often the data are used predominantly for accountability purposes that serve at national levels as a catalyst for measurement, comparison and allocation of funding (Lingard and Sellar, 2013). With increased accountability demands brought about by global competitiveness and programs for international measurement of educational attainment, there has also emerged an increase in the use of testing, which in some countries has become the dominant form of assessment. For example in Australia, national testing of students in Years 3, 5, 7 and 9 began in 2008 under the National Australia Program – Literacy and Numeracy (NAPLAN). The results from this program for each school are published on the My School website (www.myschool.edu.au), increasing the competitive nature of the testing and intensifying the demands on teachers and schools. In particular, there has been a shift in the enacted curriculum in Australia to a focus on literacy and numeracy because the curriculum is tested.

Total transcriptome, proteome, and allergome of Johnson grass pollen, which is important for allergic rhinitis in subtropical regions

Background Genomic data are lacking for many allergen sources. To circumvent this limitation, we implemented a strategy to reveal the repertoire of pollen allergens of a grass with clinical importance in subtropical regions, where an increasing proportion of the world's population resides. Objective We sought to identify and immunologically characterize the allergenic components of the Panicoideae Johnson grass pollen (JGP; Sorghum halepense). Methods The total pollen transcriptome, proteome, and allergome of JGP were documented. Serum IgE reactivities with pollen and purified allergens were assessed in 64 patients with grass pollen allergy from a subtropical region. Results Purified Sor h 1 and Sor h 13 were identified as clinically important allergen components of JGP with serum IgE reactivity in 49 (76%) and 28 (43.8%), respectively, of patients with grass pollen allergy. Within whole JGP, multiple cDNA transcripts and peptide spectra belonging to grass pollen allergen families 1, 2, 4, 7, 11, 12, 13, and 25 were identified. Pollen allergens restricted to subtropical grasses (groups 22-24) were also present within the JGP transcriptome and proteome. Mass spectrometry confirmed the IgE-reactive components of JGP included isoforms of Sor h 1, Sor h 2, Sor h 13, and Sor h 23. Conclusion Our integrated molecular approach revealed qualitative differences between the allergenic components of JGP and temperate grass pollens. Knowledge of these newly identified allergens has the potential to improve specific diagnosis and allergen immunotherapy treatment for patients with grass pollen allergy in subtropical regions and reduce the burden of allergic respiratory disease globally.

The enigma of IgE+ B-cell memory in human subjects

Our understanding of the origin and fate of the IgE-switched B cell has been markedly improved by studies in mouse models. The immediate precursor of the IgE-switched B cell is either a relatively naive nonswitched B cell or a mature IgG-switched B cell. These 2 routes are referred to as the direct and indirect pathways, respectively. IgE responses derived from each pathway differ significantly, largely reflecting the difference in time spent in a germinal center and thus time for clonal expansion, somatic hypermutation, affinity maturation, and acquisition of a memory phenotype. The clinical and therapeutic implications for IgE responses in human subjects are still a matter of debate, largely because the immunization procedures used in the animal models are significantly different from classical atopic sensitization to allergens from pollen and mites. On the basis of the limited information available, it seems likely that these atopic IgE responses are characterized by a relatively low IgG/IgE ratio, low B-cell memory, and modest affinity maturation, which fits well with the direct switching pathway. It is still unresolved how the IgE response evolves to cover a wide epitope repertoire involving many epitopes per allergen, as well as many different allergens from a single allergen source. © 2013 American Academy of Allergy, Asthma & Immunology.

Altered immunoglobulin E diversity and regulation of allergic inflammation in asthma

The clinical efficacy of anti-immunoglobulin E (IgE) therapy indicates a central role for IgE in perpetuation of allergic inflammatory diseases. Omalizumab is now uti- lized in treatment of a wide variety of allergic conditions including severe asthma, allergic rhinitis, atopic dermati- tis, food allergy and urticaria either alone or adjunct with other therapies such as steroid administration or allergen- specific immunotherapy [1, 2]. Current research activity is focused on the cellular and molecular mechanisms by which IgE influences the immunopathogenesis of allergic disease [3]. Increased knowledge of how IgE exerts its effects will underpin effective clinical use of anti-IgE treatment. In this issue Kerzel et al. [4] investigate the effects of altered antibo dy repertoire on the outcomes of an experimental model of allergic asthma.

Molecular mimicry: Can epitope mimicry induce autoimmune disease?

Mimicry of host antigens by infectious agents may induce cross-reactive autoimmune responses to epitopes within host proteins which, in susceptible individuals, may tip the balance of immunological response versus tolerance toward response and subsequently lead to autoimmune disease. Epitope mimicry may indeed be involved in the pathogenesis of several diseases such as post-viral myocarditis or Chagas disease, but for many other diseases in which it has been implicated, such as insulin-dependent diabetes mellitis or rheumatoid arthritis, convincing evidence is still lacking. Even if an epitope mimic can support a cross-reactive T or B cell response in vitro, its ability to induce an autoimmune disease in vivo will depend upon the appropriate presentation of the mimicked host antigen in the target tissue and, in the case of T cell mimics, the ability of the mimicking epitope to induce a proliferative rather than anergizing response upon engagement of the MHC-peptide complex with the T cell receptor. B cell presentation of mimicking foreign antigen to T cells is a possible mechanism for instigating an autoimmune response to self antigens that in turn can lead to autoimmune disease under particular conditions of antigen presentation, secondary signalling and effector cell repertoire. In this review evidence in support of epitope mimicry is examined in the light of the necessary immunological considerations of the theory.

Genetic associations and functional characterization of M1 aminopeptidases and immune-mediated diseases

Endosplasmic reticulum aminopeptidase 1 (ERAP1), endoplasmic reticulum aminopeptidase 2 (ERAP2) and puromycin-sensitive aminopeptidase (NPEPPS) are key zinc metallopeptidases that belong to the oxytocinase subfamily of M1 aminopeptidase family. NPEPPS catalyzes the processing of proteosome-derived peptide repertoire followed by trimming of antigenic peptides by ERAP1 and ERAP2 for presentation on major histocompatibility complex (MHC) Class I molecules. A series of genome-wide association studies have demonstrated associations of these aminopeptidases with a range of immune-mediated diseases such as ankylosing spondylitis, psoriasis, Behçet's disease, inflammatory bowel disease and type I diabetes, and significantly, genetic interaction between some aminopeptidases and HLA Class I loci with which these diseases are strongly associated. In this review, we highlight the current state of understanding of the genetic associations of this class of genes, their functional role in disease, and potential as therapeutic targets.

Repositioning the innovation rationale for public service media

The values that gave rise to the ethos of public service broadcasting (PSB) almost a century ago, and which have provided the rationale for PSBs around the world across that time, are under question. This article argues that the process of reinvention of PSBs is enhanced through repositioning the innovation rationale for public service media (PSM). It is organized around a differentiation which is part of the standard repertoire of innovation studies – that between product, process and organizational innovation – as they are being practised by the two Australian PSBs, the Australian Broadcasting Corporation (ABC) and the Special Broadcasting Service (SBS). The article then considers the general problematics of innovation for PSBs through an analysis of the operations of the public value test in the context of European PSM, and its, to this stage, non-application in Australia. The innovation rationale is argued to be a distinctive via media between complementary and comprehensive roles for PSM, which in turn suggests an international, policy-relevant research agenda focusing on international circumstances in which the public broadcaster is not market dominant.

Honey flows through fertile valleys : The cognitive and evolutionary foundations of paradise representations

The aim of the study is to explain how paradise beliefs are born from the viewpoint of mental functions of the human mind. The focus is on the observation that paradise beliefs across the world are mutually more similar than dissimilar. By using recent theories and results from the cognitive and evolutionary study of religion as well as from studies of environmental preferences, I suggest that this is because pan-human unconscious motivations, the architecture of mind, and the way the human mind processes information constrain the possible repertoire of paradise beliefs. The study is divided into two parts, theoretical and empirical. The arguments in the theoretical part are tested with data in the empirical part with two data sets. The first data set was collected using an Internet survey. The second data set was derived from literary sources. The first data test the assumption that intuitive conceptions of an environment of dreams generally follow the outlines set by evolved environmental preferences, but that they can be tweaked by modifying the presence of desirable elements. The second data test the assumption that familiarity is a dominant factor determining the content of paradise beliefs. The results of the study show that in addition to the widely studied belief in supernatural agents, belief in supernatural environments wells from the natural functioning of the human mind attesting the view that religious thinking and ideas are natural for human species and are produced by the same mental mechanisms as other cultural information. The results also help us to understand that the mental structures behind the belief in the supernatural have a wider scope than has been previously acknowledged.

Teatteri liikkeessä : Työväenteatterit Suomen teatterikentällä ja työväenliikkeessä kaksiteatterijärjestelmän syntyyn asti vuonna 1922

Workers' theatres in Finland until 1922 The topic of this dissertation is the workers' theatres in Finland before the year 1922. The main question is: why did these amateur theatres within the workers' associations become part of the professional theatre field in the 1910s by getting state subsidy as local theatre institutions? How is it possible that they received this status even after the civil war in 1918 when new professional theatres were founded all over the country? The study also asks, what kind of position did workers' theatres have in the workers' associations and in the workers' movement, what did the Social Democrats and Communists think of theatre and in particular of workers' theatre, and what kind of repertoire did the workers' theatres perform? It is a particular feature of Finland that the professional theatre field was not organised and that the workers’ movement had a relatively strong political position. The study concludes that some workers' theatres were the only steady theatre institutions in their surroundings, and thus functioned as local popular theatres performing to all social groups. Although amateur-based, they started to resemble professional theatres. Even though the Social Democratic Party did not have a specific theatre policy, the leaders of the Party appreciated and supported the workers' theatres as educational institutions and worked for their artistic improvement. The workers' theatres were also largely approved of and seen as people's theatres thought to unite and educate the nation and the working class. This reveals the need for national consensus, in the 1910s against the Russian government who worked to dissolve the autonomous position of the Finnish state, and after the civil war (1918) against the threat of a communist revolution. A wave of agitating proletarian theatre was felt in Finland in the early 1920s but it was marginalised by the large anti-communist majority.