Wolbachia uses host microRNAs to manipulate host gene expression and facilitate colonization of the dengue vector Aedes aegypti

The obligate endosymbiont Wolbachia pipientis is found in a wide range of invertebrates where they are best known for manipulating host reproduction. Recent studies have shown that Wolbachia also can modulate the lifespan of host insects and interfere with the development of human pathogens in mosquito vectors. Despite considerable study, very little is known about the molecular interactions between Wolbachia and its hosts that might mediate these effects. Using microarrays, we show that the microRNA (miRNA) profile of the mosquito, Aedes aegypti, is significantly altered by the wMelPop-CLA strain of W. pipientis. We found that a host miRNA (aae-miR-2940) is induced after Wolbachia infection in both mosquitoes and cell lines. One target of aae-miR-2940 is the Ae. aegypti metalloprotease gene. Interestingly, expression of the target gene was induced after Wolbachia infection, ectopic expression of the miRNA independent of Wolbachia, or transfection of an artificial mimic of the miRNA into mosquito cells. We also confirmed the interaction of aae-miR-2940 with the target sequences using GFP as a reporter gene. Silencing of the metalloprotease gene in both Wolbachia-infected cells and adult mosquitoes led to a significant reduction in Wolbachia density, as did inhibition of the miRNA in cells. These results indicate that manipulation of the mosquito metalloprotease gene via aae-miR-2940 is crucial for efficient maintenance of the endosymbiont. This report shows how Wolbachia alters the host miRNA profile and provides insight into the mechanisms of host manipulation used by this widespread endosymbiont.

Transient expression of Human papillomavirus type 16 L1 protein in Nicotiana benthamiana using an infectious tobamovirus vector

A Tobacco mosaic virus (TMV)-derived vector was used to express a native Human papillomavirus type 16 (HPV-16) L1 gene in Nicotiana benthamiana by means of infectious in vitro RNA transcripts inoculated onto N. benthamiana plants. HPV-16 L1 protein expression was quantitated by enzyme-linked immunosorbent assays (ELISA) after concentration of the plant extract. We estimated that the L1 product yield was 20-37 μg/kg of fresh leaf material. The L1 protein in the concentrated extract was antigenically characterised using the neutralising and conformation-specific Mabs H16:V5 and H16:E70, which bound to the plant-produced protein. Particles observed by transmission electron microscopy were mainly capsomers but virus-like particles (VLPs) similar to those produced in other systems were also present. Immunisation of rabbits with the concentrated plant extract induced a weak immune response. This is the first report of the successful expression of an HPV L1 gene in plants using a plant virus vector. © 2006 Elsevier B.V. All rights reserved.

High level protein expression in plants through the use of a novel autonomously replicating geminivirus shuttle vector

We constructed a novel autonomously replicating gene expression shuttle vector, with the aim of developing a system for transiently expressing proteins at levels useful for commercial production of vaccines and other proteins in plants. The vector, pRIC, is based on the mild strain of the geminivirus Bean yellow dwarf virus (BeYDV-m) and is replicationally released into plant cells from a recombinant Agrobacterium tumefaciens Ti plasmid. pRIC differs from most other geminivirus-based vectors in that the BeYDV replication-associated elements were included in cis rather than from a co-transfected plasmid, while the BeYDV capsid protein (CP) and movement protein (MP) genes were replaced by an antigen encoding transgene expression cassette derived from the non-replicating A. tumefaciens vector, pTRAc. We tested vector efficacy in Nicotiana benthamiana by comparing transient cytoplasmic expression between pRIC and pTRAc constructs encoding either enhanced green fluorescent protein (EGFP) or the subunit vaccine antigens, human papillomavirus subtype 16 (HPV-16) major CP L1 and human immunodeficiency virus subtype C p24 antigen. The pRIC constructs were amplified in planta by up to two orders of magnitude by replication, while 50% more HPV-16 L1 and three- to seven-fold more EGFP and HIV-1 p24 were expressed from pRIC than from pTRAc. Vector replication was shown to be correlated with increased protein expression. We anticipate that this new high-yielding plant expression vector will contribute towards the development of a viable plant production platform for vaccine candidates and other pharmaceuticals. © 2009 Blackwell Publishing Ltd.

An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6-8 kb. © 2007 Springer-Verlag.

Young children’s metarepresentational competence in data modelling

longitudinal study of data modelling across grades 1-3. The activity engaged children in designing, implementing, and analysing a survey about their new playground. Data modelling involves investigations of meaningful phenomena, deciding what is worthy of attention (identifying complex attributes), and then progressing to organising, structuring, visualising, and representing data. The core components of data modelling addressed here are children’s structuring and representing of data, with a focus on their display of metarepresentational competence (diSessa, 2004). Such competence includes students’ abilities to invent or design a variety of new representations, explain their creations, understand the role they play, and critique and compare the adequacy of representations. Reported here are the ways in which the children structured and represented their data, the metarepresentational competence displayed, and links between their metarepresentational competence and conceptual competence.

Intercultural competence in engineering education : who are we teaching?

BACKGROUND There is little doubt that our engineering graduates’ ability to identify cultural differences and their potential to impact on engineering projects, and to work effectively with these differences is of key importance in the modern engineering practice. Within engineering degree programs themselves there is also a significant need to recognise the impact of changing student and staff profiles on what happens in the classroom. The research described in this paper forms part of a larger project exploring issues of intercultural competence in engineering. PURPOSE This paper presents an observational and survey study of undergraduate and postgraduate engineering students from four institutions working in groups on tasks with a purely technical focus, or with a cultural and humanitarian element. The study sought to explore how students rate their own intercultural competence and team process and whether any differences exist depending on the nature of the task they are working on. We also investigated whether any differences were evident between groups of first year, second year and postgraduate students. DESIGN/METHOD The study used the miniCQS instrument (Ang & Van Dyne, 2008) and a Bales Interaction Process Analysis based scale (Bales, 1950; Carney, 1976) to collect students self ratings of group process, task management, and cultural experience and behaviour. The Bales IPA was also used for coding video observations of students working in groups. Survey data were used to form descriptive variables to compare outcomes across the different tasks and contexts. Observations analysed in Nvivo were used to provide commentary and additional detail on the quantitative data. RESULTS The results of the survey indicated consistent mean scores on each survey item for each group of students, despite vastly different tasks, student backgrounds and educational contexts. Some small, statistically significant mean differences existed, offering some basic insights into how task and student group composition could affect self ratings. Overall though, the results suggest minimal shift in how students view group function and their intercultural experience, irrespective of differing educational experience. CONCLUSIONS The survey results, contrasted with group observations, indicate that either students are not translating their experience (in the group tasks) into critical self assessment of their cultural competence and teamwork, or that they become more critical of team performance and cultural competence as their competence in these areas grows, so their ratings remain consistent. Both outcomes indicate that students need more intensive guidance to build their critical self and peer assessment skills in these areas irrespective of their year level of study.

Building intercultural competence one “Patch” at a time

This paper describes a program called Patches that was implemented to assist a group of Australian and Malaysian pre-service teachers to enhance their intercultural competence through their involvement in a series of reciprocal learning activities. Each learning experience was considered a “patch” that eventually created a “quilt of intercultural learning.” The purpose of this study was to enhance the intercultural competence of domestic and international students through organized intercultural activities, through a series of reflective writing sessions, and mutual engagement on a common project. The effectiveness of the Patches program was analysed in accordance with Deardorff’s elements of intercultural competence. The qualitative findings indicate that both cohorts of preservice teachers showed elements of intercultural competence through participation in the program, with both groups reporting a deeper appreciation and understanding of how to communicate more effectively in intercultural contexts.

Developing intercultural competence in pre-service teachers from Australia and Malaysia : insights from the “Patches” programme

Developing intercultural competence in pre-service teachers from Australia and Malaysia: Insights from a Patches program. Innovative pedagogies can offer pre-service teachers the opportunity to develop their intercultural competence and take up more globalised viewpoints. One such innovation is the Patches program which brought together Malaysian and Australian pre-service teachers who were studying at the same university in Brisbane, Australia, to actively explore issues of cultural and linguistic difference. The participants were 14 Australian fourth-year pre-service teachers who were enrolled in a program on inclusive education, and 58 Malaysian pre-service teachers who had recently arrived at the university in Brisbane to commence their second year of an international education program. In peer groupings, these domestic and international pre-service teachers engaged in a series of interactive tasks and reflective writing workshops exploring intercultural experiences, over a period of ten weeks. Each element or ‘patch’ in the program was designed to build up into a mosaic of intercultural learning. The flexible structuring of the Patches Program provided a supportive framework for participant interaction whilst allowing the groups to decide for themselves the nature and extent of their involvement in a series of community-related tasks. The process of negotiating and implementing these activities formed the basis for establishing meaningful relationships between the participants. The development of the participants’ intercultural competence is traced through their reflective narratives and focus group discussions, drawing on Byram’s concept of the five savoirs. Explaining aspects of Australian culture to their newly arrived Malaysian peers, allowed the Australian pre-service teachers to take a perspective of outsideness towards their own familiar social practices. In addition, being unusually positioned as the linguistic other amongst a group of Bahasa Melayu speakers, highlighted for the Australian pre-service teachers the importance of being inclusive. For the Malaysian pre-service teachers, participation in the Patches program helped to extend intercultural understandings, establish social networks with local students, and build a sense of community in their new learning environment. Both groups of pre-service teachers noted the power of “learning directly by interacting rather than through books”. In addition to interacting interculturally, the process of reflecting on these intercultural experiences is seen as integral to the development of intercultural competence.

Developing academic and intercultural competence : the impact of a Patches program

This symposium focuses on an innovative Patches program that addresses imperatives with respect to the development of intercultural competence. The term, Patches, in this project refers to writing tasks and intercultural activities wherein each task becomes a ‘patch’ that eventually creates a quilt of learning as experienced by 58 second year BEd students from Malaysia and 14 fourth year domestic (Australian) BEd students. We take intercultural competence to mean students “ability to communicate effectively and appropriately in intercultural situations based on [their] intercultural knowledge, skills and attitudes” (Deardorff, 2006). The papers in this symposium provide detailed information about the design of the program, its impact on students’ perceptions of themselves as students, writers, and emerging professionals, and students’ development of intercultural competence.

I-vector based speaker recognition using advanced channel compensation techniques

This paper investigates advanced channel compensation techniques for the purpose of improving i-vector speaker verification performance in the presence of high intersession variability using the NIST 2008 and 2010 SRE corpora. The performance of four channel compensation techniques: (a) weighted maximum margin criterion (WMMC), (b) source-normalized WMMC (SN-WMMC), (c) weighted linear discriminant analysis (WLDA), and; (d) source-normalized WLDA (SN-WLDA) have been investigated. We show that, by extracting the discriminatory information between pairs of speakers as well as capturing the source variation information in the development i-vector space, the SN-WLDA based cosine similarity scoring (CSS) i-vector system is shown to provide over 20% improvement in EER for NIST 2008 interview and microphone verification and over 10% improvement in EER for NIST 2008 telephone verification, when compared to SN-LDA based CSS i-vector system. Further, score-level fusion techniques are analyzed to combine the best channel compensation approaches, to provide over 8% improvement in DCF over the best single approach, (SN-WLDA), for NIST 2008 interview/ telephone enrolment-verification condition. Finally, we demonstrate that the improvements found in the context of CSS also generalize to state-of-the-art GPLDA with up to 14% relative improvement in EER for NIST SRE 2010 interview and microphone verification and over 7% relative improvement in EER for NIST SRE 2010 telephone verification.

Impact of Wolbachia on infection with chikungunya and yellow fever viruses in the mosquito vector Aedes aegypti

Incidence of disease due to dengue (DENV), chikungunya (CHIKV) and yellow fever (YFV) viruses is increasing in many parts of the world. The viruses are primarily transmitted by Aedes aegypti, a highly domesticated mosquito species that is notoriously difficult to control. When transinfected into Ae. aegypti, the intracellular bacterium Wolbachia has recently been shown to inhibit replication of DENVs, CHIKV, malaria parasites and filarial nematodes, providing a potentially powerful biocontrol strategy for human pathogens. Because the extent of pathogen reduction can be influenced by the strain of bacterium, we examined whether the wMel strain of Wolbachia influenced CHIKV and YFV infection in Ae. aegypti. Following exposure to viremic blood meals, CHIKV infection and dissemination rates were significantly reduced in mosquitoes with the wMel strain of Wolbachia compared to Wolbachia-uninfected controls. However, similar rates of infection and dissemination were observed in wMel infected and non-infected Ae. aegypti when intrathoracic inoculation was used to deliver virus. YFV infection, dissemination and replication were similar in wMel-infected and control mosquitoes following intrathoracic inoculations. In contrast, mosquitoes with the wMelPop strain of Wolbachia showed at least a 10(4) times reduction in YFV RNA copies compared to controls. The extent of reduction in virus infection depended on Wolbachia strain, titer and strain of the virus, and mode of exposure. Although originally proposed for dengue biocontrol, our results indicate a Wolbachia-based strategy also holds considerable promise for YFV and CHIKV suppression.

Something’s coming, something good: Identifying TPACK competence in pre-service teachers’ analyses of learning objects

There is a song at the beginning of the musical, West Side Story, where the character Tony sings that “something’s coming, something good.” The song is an anthem of optimism, brimming with promise. This paper is about the long-held promise of information and communication technology (ICT) to transform teaching and learning, to modernise the learning environment of the classroom, and to create a new digital pedagogy. But much of our experience to date in the schooling sector tells more of resistance and reaction than revolution, of more of the same but with a computer in the corner and of ICT activities as unwelcome time-fillers/time-wasters. Recently, a group of pre-service teachers in a postgraduate primary education degree in an Australian university were introduced to learning objects in an ICT immersion program. Their analyses and related responses, as recorded in online journals, have here been interpreted in terms of TPACK (Technological Pedagogical and Content Knowledge). Against contemporary observation, these students generally displayed high levels of competence and highly positive dispositions of students to the integration of ICT in their future classrooms. In short, they displayed the same optimism and confidence as the fictional “Tony” in believing that something good was coming.

Assessing testamentary and decision-making competence/capacity in the Australian context

Australia lacks a satisfactory, national paradigm for assessing competence and capacity in the context of testamentary, enduring power of attorney and advance care directive documents. Competence/capacity assessments are currently conducted on an ad hoc basis by legal and/or medical professionals. The reliability of the assessment process is subject to the skill set and mutual understanding of the legal and/or medical professional conducting the assessment. There is a growth in the prevalence of diseases such as dementia. Such diseases impact upon cognition which increasingly necessitates collaboration between the legal and medical professions when assessing the effect of mentally disabling conditions upon competency/capacity. Miscommunication and lack of understanding between legal and medical professionals involved could impede the development of a satisfactory paradigm. A qualitative study seeking the views of legal and medical professionals who practise in this area has been conducted. This incorporated surveys and interviews of 10 legal and 20 medical practitioners. Some of the results are discussed here. Practitioners were asked whether there is a standard approach and whether national guidelines were desirable. There was general agreement that uniform guidelines for the assessment of competence/capacity would be desirable. The interviews also canvassed views as to the state of the relationship between the professions. The results of the empirical research support the hypothesis that relations between the professions could be improved. The development of a national paradigm would promote consistency and transparency of process, helping to improve the professional relationship and maximising the principles of autonomy, participation and dignity.

Improving short utterance based I-vector speaker recognition using source and utterance-duration normalization techniques

A significant amount of speech is typically required for speaker verification system development and evaluation, especially in the presence of large intersession variability. This paper introduces a source and utterance duration normalized linear discriminant analysis (SUN-LDA) approaches to compensate session variability in short-utterance i-vector speaker verification systems. Two variations of SUN-LDA are proposed where normalization techniques are used to capture source variation from both short and full-length development i-vectors, one based upon pooling (SUN-LDA-pooled) and the other on concatenation (SUN-LDA-concat) across the duration and source-dependent session variation. Both the SUN-LDA-pooled and SUN-LDA-concat techniques are shown to provide improvement over traditional LDA on NIST 08 truncated 10sec-10sec evaluation conditions, with the highest improvement obtained with the SUN-LDA-concat technique achieving a relative improvement of 8% in EER for mis-matched conditions and over 3% for matched conditions over traditional LDA approaches.

Building project competence : developing dynamic capabilities with knowledge governance mechanisms

This research investigates the development of project competence, and particularly, three related dynamic capabilities (shifting, adapting, leveraging) that contribute to project competence development. In doing so, we make use of the emerging literature on knowledge governance and theorize how knowledge governance mechanisms can move the organization towards desired knowledge-based goals. A multiple-case study comprising 23 cases advances our understanding of the elements that trigger, enable, hamper, and drive shifting, leveraging and adapting. Finally, the paper offers a tentative framework of dynamic capability building promoting project competence development.