Translation of the flavivirus Kunjin NS3 gene in cis but not its RNA sequence or secondary structure is essential for efficient RNA packaging

Our previous studies using trans-complementation analysis of Kunjin virus (KUN) full-length cDNA clones harboring in-frame deletions in the NS3 gene demonstrated the inability of these defective complemented RNAs to be packaged into virus particles (W. J. Liu, P. L. Sedlak, N. Kondratieva, and A. A. Khromykh, J. Virol. 76:10766-10775). In this study we aimed to establish whether this requirement for NS3 in RNA packaging is determined by the secondary RNA structure of the NS3 gene or by the essential role of the translated NS3 gene product. Multiple silent mutations of three computer-predicted stable RNA structures in the NS3 coding region of KUN replicon RNA aimed at disrupting RNA secondary structure without affecting amino acid sequence did not affect RNA replication and packaging into virus-like particles in the packaging cell line, thus demonstrating that the predicted conserved RNA structures in the NS3 gene do not play a role in RNA replication and/or packaging. In contrast, double frameshift mutations in the NS3 coding region of full-length KUN RNA, producing scrambled NS3 protein but retaining secondary RNA structure, resulted in the loss of ability of these defective RNAs to be packaged into virus particles in complementation experiments in KUN replicon-expressing cells. Furthermore, the more robust complementation-packaging system based on established stable cell lines producing large amounts of complemented replicating NS3-deficient replicon RNAs and infection with KUN virus to provide structural proteins also failed to detect any secreted virus-like particles containing packaged NS3-deficient replicon RNAs. These results have now firmly established the requirement of KUN NS3 protein translated in cis for genome packaging into virus particles.

Minimal mutation of the cytoplasmic tail inhibits the ability of E-cadherin to activate Rac but not phosphatidylinositol 3-kinase - Direct evidence of a role for cadherin-activated Rac signaling in adhesion and contact formation

Classic cadherins are adhesion-activated cell signaling receptors. In particular, homophilic cadherin ligation can directly activate Rho family GTPases and phosphatidylinositol 3-kinase (PI3-kinase), signaling molecules with the capacity to support the morphogenetic effects of these adhesion molecules during development and disease. However, the molecular basis for cadherin signaling has not been elucidated, nor is its precise contribution to cadherin function yet understood. One attractive hypothesis is that cadherin-activated signaling participates in stabilizing adhesive contacts ( Yap, A. S., and Kovacs, E. M. ( 2003) J. Cell Biol. 160, 11-16). We now report that minimal mutation of the cadherin cytoplasmic tail to uncouple binding of p120-ctn ablated the ability of E-cadherin to activate Rac. This was accompanied by profound defects in the capacity of cells to establish stable adhesive contacts, defects that were rescued by sustained Rac signaling. These data provide direct evidence for a role of cadherin-activated Rac signaling in contact formation and adhesive stabilization. In contrast, cadherin-activated PI3-kinase signaling was not affected by loss of p120-ctn binding. The molecular requirements for E-cadherin to activate Rac signaling thus appear distinct from those that stimulate PI3-kinase, and we postulate that p120-ctn may play a central role in the E-cadherin-Rac signaling pathway.

Pretend play, mirror self-recognition and imitation: A longitudinal investigation through the second year

The aim in the current study was to investigate the emergence of pretend play, mirror self-recognition, synchronic imitation and deferred imitation in normally developing human infants. A longitudinal study was conducted with 98 infants seen at three-monthly intervals from 12 through to 24 months of age. At each session the infants were tested on a range of tasks assessing the four target skills. Deferred imitation was found to emerge prior to synchronic imitation, pretend play and mirror self-recognition. In contrast, the latter three skills emerged between 18 and 21 months and followed similar developmental trajectories. Deferred imitation was found to hold a prerequisite relation with these three skills. Synchronic imitation, pretend play and mirror self-recognition were not closely associated and no prerequisite relations were found between these skills. These findings are discussed in the context of current theories regarding the development of pretend play, mirror self-recognition, synchronic imitation and deferred imitation in the second year. (C) 2004 Elsevier Inc. All rights reserved.

Play it again: did this melody occur more frequently or was it heard more recently? The role of stimulus familiarity in episodic recognition of music

Episodic recognition of novel and familiar melodies was examined by asking participants to make judgments about the recency and frequency of presentation of melodies over the course of two days of testing. For novel melodies, recency judgments were poor and participants often confused the number of presentations of a melody with its day of presentation; melodies heard frequently were judged as have been heard more recently than they actually were. For familiar melodies, recency judgments were much more accurate and the number of presentations of a melody helped rather than hindered performance. Frequency judgments were generally more accurate than recency judgments and did not demonstrate the same interaction with musical familiarity. Overall, these findings suggest that (1) episodic recognition of novel melodies is based more on a generalized feeling of familiarity than on a specific episodic memory, (2) frequency information contributes more strongly to this generalized memory than recency information, and (3) the formation of an episodic memory for a melody depends either on the overall familiarity of the stimulus or the availability of a verbal label. (C) 2004 Elsevier B.V. All rights reserved.

Editorial

The article presents information on the various papers published in the December 2005 issue of the periodical "Australian Journal of Communication." In one of the papers, author Jane Johnston updates her earlier work on communication in the Australian court system by examining the courts' communication with the media. Chika Anyanwu makes a contribution to the literature on diasporic discourses by explaining the ways in which the new media technologies have redefined diaspora by enabling diasporic citizens to connect with their homelands. In their paper, Mark Balnaves and Kim Tomlinson-Baillie outline strategies that the international games industry brings to play when developing games, allowing children to participate in and change the narrative as it progresses through to a new world.

Ebola virus glycoprotein GP is not cytotoxic when expressed constitutively at a moderate level

Transient expression of Ebola virus (EBOV) glycoprotein GP causes downregulation of surface proteins, cell rounding and detachment, a phenomenon believed to play a central role in the pathogenicity of the virus. In this study, evidence that moderate expression of GP does not result in such morphological changes was provided. It was shown that GP continuously produced in 293T cells from the Kunjin virus replicon was correctly processed and transported to the plasma membrane without affecting the surface expression of beta 1 and alpha 5 integrins and major histocompatibility complex I molecules. The level of GIP expression in Kunjin replicon GP-expressing cells was similar to that observed in cells infected with EBOV early in infection and lower than that produced in cells transfected with plasmid DNA, phCMV-GP(1) expressing GP from a strong promoter. Importantly, transient transfection of Kunjin replicon GIP-expressing cells with GIP-coding plasmid DNA resulted in overexpression of GP, which lead to the downregulation of surface molecules and massive rounding and detachment of transfected cells. Here, it was also demonstrated that cell rounding and downregulation of the surface markers are the late events in EBOV infection, whereas synthesis and massive release of virus particles occur at early steps and do not cause significant cytotoxic effects. These findings indicate that the synthesis of EBOV GP in virus-infected cells is controlled well by several mechanisms that do not allow GP overexpression and hence the early appearance of its cytotoxic properties.

Maintaining human agency in the design on context-aware systems: Design games in a dental surgery

This paper describes a series of design games, specifically aimed at exploring shifts in human agency in order to inform the design of context-aware applications. The games focused on understanding information handling issues in dental practice with participants from a university dental school playing an active role in the activities. Participatory design activities help participants to reveal potential implicit technical resources that can be presented explicitly in technologies in order to assist humans in managing their interactions with and amidst technical systems gracefully.

It's all in the game: Teaching software process concepts

A major challenge in teaching software engineering to undergraduates is that most students have limited industry experience, so the problems addressed are unknown and hence unappreciated. Issues of scope prevent a realistic software engineering experience, and students often graduate with a simplistic view of software engineering’s challenges. Problems and Programmers (PnP) is a competitive, physical card game that simulates the software engineering process from requirements specification to product delivery. Deliverables are abstracted, allowing a focus on process issues and for lessons to be learned in a relatively short time. The rules are easy to understand and the game’s physical nature allows for face-to-face interaction between players. The game’s developers have described PnP in previous publications, but this paper reports the game’s use within a larger educational scheme. Students learn and play PnP, and then are required to create a software requirements specification based on the game. Finally, students reflect on the game’s strengths and weaknesses and their experiences in an individual essay. The paper discusses this approach, students’ experiences and overall outcomes, and offers an independent, critical look at the game, its use, and potential improvements.

Suit keen renovator: Alternate reality design

Virtual territories and their theme parks are more akin to the physical world of real estate than they might at first appear. The trick in triggering the designer's imagination, is to find a 'nice renovator' (cottage/ house) at a low price, with loads of potential, and by doing it on the cheap to add character, and engage the imagination. Here the designer can construct changes from an imagined space. Vision is more important than how the actual place presents.This work describes a case study involving undergraduate students in the Creative Industries who needed a place to explore, so as to create their own visions and projects. The place had to inspire, trigger engagement, and their imaginations. At the same time it was important that the place did not coerce activity, or distract from the task by confusing tools with task, or architectural navigation with conceptual skills.The solution was an alternate reality.