Serpin overexpression in Plasmodium-infected midgut cells

Summary The design of effective, vector-based malaria transmission blocking strategies relies on a thorough understanding of the molecular and cellular interactions that occur during the parasite sporogonic cycle in the mosquito. During Plasmodium berghei invasion, transcription from the SRPN10 locus, encoding four serine protease inhibitors of the ovalbumin family, is strongly induced in the mosquito midgut. Herein we demonstrate that intense induction as well as redistribution of SRPN10 occurs specifically in the parasite-invaded midgut epithelial cells. Quantitative analysis establishes that in response to epithelial invasion, SRPN10 translocates from the nucleus to the cytoplasm and this is followed by strong SRPN10 overexpression. The invaded cells exhibit signs of apoptosis, suggesting a link between this type of intracellular serpin and epithelial damage. The SRPN10 gene products constitute a novel, robust and cell-autonomous marker of midgut invasion by ookinetes. The SRPN10 dynamics at the subcellular level confirm and further elaborate the 'time bomb' model of P. berghei invasion in both Anopheles stephensi and Anopheles gambiae. In contrast, this syndrome of responses is not elicited by mutant P. berghei ookinetes lacking the major ookinete surface proteins, P28 and P25. Molecular markers with defined expression patterns, in combination with mutant parasite strains, will facilitate dissection of the molecular mechanisms underlying vector competence and development of effective transmission blocking strategies

KP-LAB Knowledge Practices Laboratory -- External release of end-user applications

deliverablesThis deliverable describes the M24 release of the End user applications for knowledge practices software v2.0.0. The deliverable includes the technical development performed until M24 (January 2008) within WP6 according to Description of Work 2.1 and D6.4 M21 specification of end-user applications. The current release is comprised of two set of tools: 1. Shared Space Tool The shared space and the accompanying support material can be found on the Internet at: http://2d.mobile.evtek.fi:8080/shared-space 2. Map-It. The installer program for Map-It v2.0.0 is available at: http://www.kp-lab.org/intranet/testable-tools/kp-lab-tools/map-it/map-it-2-0.0 Please consult the "Getting Started" Note before installing and using Map-It: http://www.kp-lab.org/intranet/testable-tools/kp-lab-tools/map-it/getting-started-with-map-it 3. Change Laboratory tools The release targeted for the end users participating in the trials planned to be conducted in the CL Working Knot can be accessed via the following link: http://2d.mobile.evtek.fi:8080/shared-space/cl.html Anyone who wishes to try the software out but is not participating in the Change Laboratory trials should use the development deployment on: http://mielikki.mobile.evtek.fi/shared-space/cl.html The M24 release of Semantic Multimedia Annotation tools is still delayed. The release of CASS Memo Client has been postponed to be included in the M28 release in DoW3

Toward Anopheles transformation: Minos element activity in anopheline cells and embryos

The ability of the Minos transposable element to function as a transformation vector in anopheline mosquitoes was assessed. Two recently established Anopheles gambiae cell lines were stably transformed by using marked Minos transposons in the presence of a helper plasmid expressing transposase. The markers were either the green fluorescent protein or the hygromycin B phosphotransferase gene driven by the Drosophila Hsp70 promoter. Cloning and sequencing of the integration sites demonstrated that insertions in the cell genome occurred through the action of Minos transposase. Furthermore, an interplasmid transposition assay established that Minos transposase is active in the cytoplasmic environment of Anopheles stephensi embryos: interplasmid transposition events isolated from injected preblastoderm embryos were identified as Minos transposase-mediated integrations, and no events were recorded in the absence of an active transposase. These results demonstrate that Minos vectors are suitable candidates for germ-line transformation of anopheline mosquitoes

KP-LAB Knowledge Practices Laboratory -- Release of end-user tools

deliverablesThis deliverable describes the releases of KP-Lab end user applications and tools made during the DoW4 period of the project (M37-M48). These are as follows: KPE: Shared Space Views and Common, Support and Optional Tools provide the major functionality for the Knowledge Practices Environment. The Shared Space Views visualize the knowledge artefacts and their relations from different perspective, allowing users to view and access the information contained in a shared space in flexible manners. Common tools refer to the tightly integrated tools of KPE, which are available inside a shared space for working with knowledge artefacts. The support tools provide generic supplementary functionality to Shared Space Views and other KP-Lab tools, such as awareness, search, help as well as preference and settings. The optional tools provide functionality to support some specific aspects of or types of knowledge creation processes, such as real-time collaborative writing, activity system design, multimedia annotation and visual modelling. KPE Analytic tools provide means for two main categories of analytic facilities: 1) Data export tool (DE) for automatic data collection for its analysis in any third party tools and 2) Timeline-based analyzer (TLBA) and Visual analyzer (VA) for integrated reflection on knowledge creation processes and their analysis. * Data Export tool allows researchers and teachers to extract summary tables of user activities from the KPE for on-line investigation, and to export them for elaborations with any third party tool for analysis. In order to bring this tool closer to casual users (students, teachers or workers), the visual presentation of relations between KPE users has been added in the last period. * Visual analyzer allows users to analyze participation and activities within past or ongoing knowledge creation processes, by visually representing them based on information stored in the produced logs. More precisely, it visualizes frequencies of object-related activities in KPE and provides detailed information on the nature and type of the activities performed on particular (types of) knowledge objects. These visualizations stimulate teachers and students to reflect on the distribution and types of their activities with respect to time, type of object or subject etc. * Timeline-based analyzer allows users to display chronologically events that were recorded by the KPE tools, to define and store possible external events which could not have been recorded by the KPE tools and to define patterns of actions that can be identified in the historical data. In contrast to Visual Analyzer and Data Export tools, TLBA brings chronological overview of user actions into the user interface, which enables users to see and explore what kind of activities were performed on certain object in the shared space of interest. Activity System Design Tools (ASDT) enable users to look to the history, present and future of their work activity in a way that helps address issues critical for deliberate transformation of prevailing practices (according to the Change Laboratory intervention method). ASDT is a plug-in to Knowledge Practices Environment, utilizing its views and functionalities. In addition, ASDT has a specific view, Virtual whiteboard, which is designed based on the key elements of a developmental work research process and its conceptual tools. Semantic Multimedia Annotation Tool (SMAT) is a rich internet application that facilitates an individuals or a groups activity of assigning annotations to the document fragments. The tools allow users to plan and organise their annotation activity, structure any multimedia document by dividing it into fragments, annotate the document formally, informally or by linking external documents to specific anchors, as well as to analyse structurally and statistically annotations and visualize analysis results. SMAT is adaptable to the users domain. Meeting Support Tools propose new approaches to meeting practices. They support the preparation, execution and analysis of meetings (face-to-face and remote) by capturing synchronous and asynchronous interactions through the collaborative elaboration of "discussion maps". Map-It allows the use of meeting templates, advance individual preparations, share of artifacts, planning and follow-up of actions, automatic generation of meeting minutes in various formats. M2T, a rich internet application, provides analytical facilities for exploring meeting practices instrumented using Map-It and their integration in larger-scope activities, by connecting them to other KP-Lab concepts, visualisations (KPEs Content Views) and tools (ToDo). This deliverable provides the general description of tools in terms of targeted users, requirements for the use, new features, known issues, as well as deviations from the DII.8 specifications. More details can be found through the material available in the project intranet at: http://www.kp-lab.org/intranet/testable-tools/kp-lab-tools