Origin and evolution of 3'UTR of flaviviruses: long direct repeats as a basis for the formation of secondary structures and their significance for virus transmission

The 3' untranslated regions (3'UTRs) of flaviviruses are reviewed and analyzed in relation to short sequences conserved as direct repeats (DRs). Previously, alignments of the 3'UTRs have been constructed for three of the four recognized flavivirus groups, namely mosquito-borne, tick-borne, and nonclassified flaviviruses (MBFV, TBFV, and NCFV, respectively). This revealed (1) six long repeat sequences (LRSs) in the 3'UTR and open-reading frame (ORF) of the TBFV, (2) duplication of the 3'UTR of the NCFV by intramolecular recombination, and (3) the possibility of a common origin for all DRs within the MBFV. We have now extended this analysis and review it in the context of all previous published analyses. This has been achieved by constructing a robust alignment between all flaviviruses using the published DRs and secondary RNA structures as "anchors" to reveal additional homologies along the 3'UTR. This approach identified nucleotide regions within the MBFV, NKV (no-known vector viruses), and NCFV 3'UTRs that are homologous to different LRSs in the TBFV 3'UTR and ORF. The analysis revealed that some of the DRs and secondary RNA structures described individually within each flavivirus group share common evolutionary origins. The 3'UTR of flaviviruses, and possibly the ORF, therefore probably evolved through multiple duplication of an RNA domain, homologous to the LRS previously identified only in the TBFV. The short DRs in all virus groups appear to represent the evolutionary remnants of these domains rather than resulting from new duplications. The relevance of these flavivirus DRs to evolution, diversity, 3'UTR enhancer function, and virus transmission is reviewed.

Transmission properties of Iranian wheat stripe virus

Transmission properties of Iranian wheat stripe virus (IWSV), a tentative member of the genus Tenuivirus, were studied. Results showed that similar to other tenuiviruses, IWSV multiplies in its vector, Unkanodes tanasijevici. In bioassay experiments, IWSV transmission rate by individual U. tanasijevici showed an increase with time after acquisition. IWSV was transovarially transmitted to 88-100% of progeny. The nymphs continued to be infective in the adult stage but with decreased efficiency. Males and females transmitted the virus with equal efficiency. Transmission properties of IWSV confirm the position of the virus in the genus Tenuivirus.

Evaluation of malacosporean life cycles through transmission studies

Myxozoans, belonging to the recently described Class Malacosporea, parasitise freshwater bryozoans during at least part of their life cycle, but no complete malacosporean life cycle is known to date. One of the 2 described malacosporeans is Tetracapsuloides bryosalmonae, the causative agent of salmonid proliferative kidney disease. The other is Buddenbrockia plumatellae, so far only found in freshwater bryozoans. Our investigations evaluated malacosporean life cycles, focusing on transmission from fish to bryozoan and from bryozoan to bryozoan. We exposed bryozoans to possible infection from: stages of T bryosalmonae in fish kidney and released in fish urine; spores of T bryosalmonae that had developed in bryozoan hosts; and spores and sac stages of B. plumatellae that had developed in bryozoans. Infections were never observed by microscopic examination of post-exposure, cultured bryozoans and none were detected by PCR after culture. Our consistent negative results are compelling: trials incorporated a broad range of parasite stages and potential hosts, and failure of transmission across trials cannot be ascribed to low spore concentrations or immature infective stages. The absence of evidence for bryozoan to bryozoan transmissions for both malacosporeans strongly indicates that such transmission is precluded in malacosporean life cycles. Overall, our results imply that there may be another malacosporean host which remains unidentified, although transmission from fish to bryozoans requires further investigation. However, the highly clonal life history of freshwater bryozoans is likely to allow both long-term persistence and spread of infection within bryozoan populations, precluding the requirement for regular transmission from an alternate host.

Impact damage characterisation of thermoplastic matrix composites using transmission transient thermography

In this work, IR thermography is used as a non-destructive tool for impact damage characterisation on thermoplastic E-glass/polypropylene composites for automotive applications. The aim of this experimentation was to compare impact resistance and to characterise damage patterns of different laminates, in order to provide indications for their use in components. Two E-glass/polypropylene composites, commingled ®Twintex (with three different weave structures: directional, balanced and 3-D) and random reinforced GMT, were in particular characterised. Directional and balanced Twintex were also coupled in a number of hybrid configurations with GMT to evaluate the possible use of GMT/Twintex hybrids in high-energy absorption components. The laminates were impacted using a falling weight tower, with impact energies ranging from 15 J to penetration. Using IR thermography during cooling down following a long pulse (3 s), impact damaged areas were characterised and the influence of weave structure on damage patterns was studied. IR thermography offered good accuracy for laminates with thickness not exceeding 3.5 mm: this appears to be a limit for the direct use of this method on components, where more refined signal treatment would probably be needed for impact damage characterisation.

When school is not enough: new initiatives in intergenerational language transmission in Wales

This paper examines language reproduction in the family in the context of a highly innovative project in Wales, where the Welsh language has been in decline for over a century. Although Welsh-medium schooling has played a pivotal role in slowing and even reversing language shift in recent decades, there is mounting evidence of the dangers of over-reliance on education. The Twf (Growth) Project was established in 2002 with funding from the National Assembly for Wales with the aim of raising awareness of the benefits of bilingualism among parents and prospective parents. Analysis of interviews with the main stakeholders in the project (managers, the Twf project officers, parents, health workers and a range of other partners), publicity materials and observations of project staff at work suggests that the achievements of the project lie in two main areas: the recognition of the need for building strong alliances with professional groups and organisations that work with families with young children; and the development of a marketing strategy appropriate for the target audience. It is argued that the experience of the project will be of interest to those addressing the issue of intergenerational transmission in a range of other minority language settings.

Language transmission in the family in Wales: an example of innovative language planning.

This paper examines the achievements to date of Twf (“Growth”) — a project initiated as part of language planning efforts in Wales to encourage families to bring up their children to be bilingual. Evidence is presented of the ways in which the project has succeeded in raising awareness of the advantages of bilingualism amongst parents, prospective parents and the public at large by working strategically with health professionals and Early Years organizations, and by developing a range of highly innovative promotional materials. Given the central importance of the family as a site of intergenerational language transmission, the achievements of this project are likely to be of interest to those concerned with language planning in other minority communities in many other parts of the world. The lessons for language planning both in Wales and in other settings are discussed.