We present the results of a process to attempt to identify 100 questions that, if answered, would make a substantial difference to terrestrial and marine landscape restoration in Europe. Representatives from a wide range of European governmental and non-governmental conservation organisations, universities, independent ecologists and land managers compiled 677 questions relating to all aspects of European landscape restoration for nature and people. The questions were shortlisted by an email vote, followed by a two-day workshop, to produce the final list of 100 questions. Many of the final questions evolved through a process of modification and combination as the workshop progressed. The questions are divided into eight sections: conservation of biodiversity; connectivity, migration and translocations; delivering and evaluating restoration; natural processes; ecosystem services; social and cultural aspects of restoration; policy and governance; and economics. We anticipate that these questions will help identify new directions for researchers and policy-makers and assist funders and programme managers in allocating funds and planning projects, resulting in improved understanding and implementation of landscape-scale ecological restoration in Europe.

Tomato spotted wilt virus (TSWV) is an economically important viral pathogen of a wide range of field and horticultural crops. We developed an artificial microRNA (amiRNA) strategy against TSWV, targeting the nucleoprotein (N) and silencing suppressor (NSs) genes. The amiRNA constructs replaced the natural miRNA in a shortened Arabidopsis 173-nucleotide (nt) miR159a precursor backbone (athmiR159a) without the stem base extending beyond the miR/miR* duplex. Further, each amiRNA was modified to contain a mismatch (wobble) sequence at nucleotide position 12 and 13 on the complementary strand amiRNA*, mimicking the endogenous miR159a sequence structure. Transient expression in Nicotiana benthamiana demonstrated that the introduction of a wobble sequence did not alter amiRNA expression levels. Following challenge inoculation with TSWV, plants expressing N-specific amiRNAs with or without the wobble remained asymptomatic and were negative for TSWV by ELISA. In contrast, plants expressing the NSs-specific amiRNAs were symptomatic and accumulated high levels of TSWV. Similar findings were obtained in stably transformed Nicotiana tabacum plants. Our results show that a shortened 173-nt athmiR159a backbone is sufficient to express amiRNAs and that the presence of mismatch at position 12-13 does not influence amiRNA expression or conferring of resistance. We also show that selection of target gene and positional effect are critical in amiRNA-based approach for introducing resistance. These findings open the possibility of employing the amiRNA approach for broad-spectrum resistance to tospoviruses as well as other viruses. (C) 2015 Elsevier B.V. All rights reserved.

Summary 1. The conservation of biodiversity depends upon both policy and regulatory frameworks. Here, we identify priority policy developments that would support conservation in the UK in the light of technological developments, changes in knowledge or environmental change. 2. A team of seven representatives from governmental organizations, 17 from non‐governmental organizations and six academics provided an assessment of the priority issues. The representatives consulted widely and identified a long‐list of 117 issues. 3. Following voting and discussion during a 2‐day meeting, these were reduced to a final list of 25 issues and their potential policy options and research needs were identified. Many of the policies related to recent changes in approaches to conservation, such as increased interest in ecosystem services, adaptation to climate change and landscape ecology. 4. We anticipate that this paper will be useful for policy makers, nature conservation delivery agencies, the research community and conservation policy advocates. 5. Although many of the options have global significance, we suggest that other countries consider an equivalent exercise. We recommend that such an exercise be carried out in the UK at regular intervals, say every 5 years, to explore how biodiversity conservation can best be supported by linked policy development and research in a changing world. 6. Synthesis and applications. Opportunities for policy development were prioritized and for each of the top 25 we identified the current context, policy options and research questions. These largely addressed new issues relating to developing topics such as ecosystem services, landscape planning and nanotechnology. We envisage that this will largely be used by researchers wishing to make a contribution to potential policy debates.

Bovine herpesvirus 1 (BoHV-1) is a ubiquitous and important pathogen of cattle worldwide. This study reports the identification of 10 microRNA (miRNA) genes, Bhv1-mir-B1-Bhv1-mir-B10, encoded by the BoHV-1 genome that were processed into 12 detectable mature miRNAs as determined by ultra-high throughput sequencing bioinformatics analyses of small RNA libraries and expression studies. We found that four of the miRNA genes were present as two copies in the BoHV-1 genome, resulting in a total of 14 miRNA encoding loci. Unique features of the BoHV-1 miRNAs include evidence of bidirectional transcription and a close association of two miRNA genes with the origin of replication, including one miRNA that is encoded within the origin of replication. The miRNA gene Bhv1-mir-B5 was encoded on the opposite DNA strand to the latency associated transcript, potentially giving rise to antisense transcripts originating from this locus. The association of herpesvirus miRNAs with latency appears to be a common feature in the alphaherpesviruses. Analyses of the BoHV-5 genome for putative miRNA gene orthologues identified a high degree of evolutionary conservation for nine of the BoHV-1 miRNA genes. The possible roles for BoHV-1 miRNAs in the regulation of known BoHV-1 transcription units and the genetics of the BoHV-1 genotypes are also discussed.

Summary Horizon scanning is an essential tool for environmental scientists if they are to contribute to the evidence base for Government, its agencies and other decision makers to devise and implement environmental policies. The implication of not foreseeing issues that are foreseeable is illustrated by the contentious responses to genetically modified herbicide‐tolerant crops in the UK, and by challenges surrounding biofuels, foot and mouth disease, avian influenza and climate change. A total of 35 representatives from organizations involved in environmental policy, academia, scientific journalism and horizon scanning were asked to use wide consultation to identify the future novel or step changes in threats to, and opportunities for, biodiversity that might arise in the UK up to 2050, but that had not been important in the recent past. At least 452 people were consulted. Cases for 195 submitted issues were distributed to all participants for comments and additions. All issues were scored (probability, hazard, novelty and overall score) prior to a 2‐day workshop. Shortlisting to 41 issues and then the final 25 issues, together with refinement of these issues, took place at the workshop during another two rounds of discussion and scoring. We provide summaries of the 25 shortlisted issues and outline the research needs. We suggest that horizon scanning incorporating wide consultation with providers and users of environmental science is used by environmental policy makers and researchers. This can be used to identify gaps in knowledge and policy, and to identify future key issues for biodiversity, including those arising from outside the domains of ecology and biodiversity. Synthesis and applications. Horizon scanning can be used by environmental policy makers and researchers to identify gaps in knowledge and policy. Drawing on the experience, expertise and research of policy advisors, academics and journalists, this exercise helps set the agenda for policy, practice and research.

Transgenic tobacco plants, carrying a Potato virus Y (PVY)-NIa hairpin sequence separated by a unique unrelated spacer sequence were specifically silenced and highly resistant to PVY infection. In such plants neither PVY-NIa nor spacer transgene transcripts were detectable by specific quantitative real time reverse transcriptase PCR (RT-qPCR) assays of similar relative efficiencies developed for direct comparative analysis. However, small interfering RNAs (siRNAs) specific for the PVY sequence of the transgene and none specific for the LNYV spacer sequence were detected. Following infection with Cucumber mosaic virus (CMV), which suppresses dsRNA-induced RNA silencing, transcript levels of PVY-NIa as well as spacer sequence increased manifold with the same time course. The cellular abundance of the single-stranded (ss) spacer sequence was consistently higher than that of PVY dsRNA in all cases. The results show that during RNA silencing and its suppression of a hairpin transcript in transgenic tobacco, the ssRNA spacer sequence is affected differently than the dsRNA. In PVY-silenced plants, the spacer is efficiently degraded by a mechanism not involving the accumulation of siRNAs, while following suppression of RNA silencing by CMV, the spacer appears protected from degradation.

New research is beginning to illuminate the relationship between hedge funds and systemic risk. Could a hedge fund crisis plunge capital markets into turmoil?

Gene transfer by intramuscular injection of plasmid DNA has potential application in gene therapy. We examined factors affecting the number of expressing fibers, in contrast to total expression, following injection of plasmid DNA. Barium chloride proved effective in inducing muscle necrosis and regeneration in mice, and this increased the number of fibers expressing a reporter gene. Coinjection of ion-channel modulators did not increase the number of positive fibers, but increasing dose and repeated administration of plasmid did. Importantly, the plasmid size (7-16 kb) did not affect the number of fibers expressing the transgene, in both normal and regenerating muscle.

Albendazole resistance was induced in three different Giardia cultures following growth in successively increasing amounts of drug. One of the lines was previously resistant to high levels of metronidazole and was able to grow in 2 microM albendazole. The other two survived exposure to 0.8 microM, while normally lethal levels of albendazole against Giardia in vitro were around 0.1-0.2 microM. Albendazole-resistant Giardia were cross-resistant to parbendazole. Major chromosome rearrangements were evident in the line resistant to 2 microM albendazole and IFA with antitubulin antibody indicated differences in the cytoskeleton, particularly the median body, between sensitive and resistant lines. This implicates the cytoskeleton in the mechanism of resistance. Substitution of Tyr for Phe is a consistent beta-tubulin amino acid change in the benzimidazole-resistant helminths and fungi so far analyzed. PCR primers were designed from the published Giardia beta-tubulin gene sequence and spanned the region encoding Phe at position 200. Sequence data from albendazole-resistant Giardia demonstrated that the beta-tubulin gene did not carry a mutation in the codon for amino acid 200. These data suggest that Phe at position 200 in beta-tubulin is not necessary for benzimidazole resistance.

Journal Article Identifying training and technical assistance needs in community coalitions: a developmental approach Get access Paul Florin, Paul Florin Community Research and Services Team, Department of Psychology, University of Rhode IslandKingston, RI, USA Search for other works by this author on: Oxford Academic PubMed Google Scholar Roger Mitchell, Roger Mitchell Community Research and Services Team, Department of Psychology, University of Rhode IslandKingston, RI, USA Search for other works by this author on: Oxford Academic PubMed Google Scholar John Stevenson John Stevenson Community Research and Services Team, Department of Psychology, University of Rhode IslandKingston, RI, USA Search for other works by this author on: Oxford Academic PubMed Google Scholar Health Education Research, Volume 8, Issue 3, September 1993, Pages 417–432, https://doi.org/10.1093/her/8.3.417 Published: 01 September 1993

Journal Article Complete nucleotide sequence of the ribosomal RNA tandem repeat unit from Giardia intestinalis Get access Andrew Healey, Andrew Healey Queensland Institute of Medical ResearchBrisbane, Queensland 4006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Roger Mitchell, Roger Mitchell Queensland Institute of Medical ResearchBrisbane, Queensland 4006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Jacqueline A. Upcroft, Jacqueline A. Upcroft Queensland Institute of Medical ResearchBrisbane, Queensland 4006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Peter F. L. Boreham, Peter F. L. Boreham Queensland Institute of Medical ResearchBrisbane, Queensland 4006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Peter Upcroft Peter Upcroft * Queensland Institute of Medical ResearchBrisbane, Queensland 4006, Australia * To whom correspondence should be addressed Search for other works by this author on: Oxford Academic PubMed Google Scholar Nucleic Acids Research, Volume 18, Issue 13, 11 July 1990, Page 4006, https://doi.org/10.1093/nar/18.13.4006 Published: 11 July 1990

AbstractImproved teaching techniques are essential for integrating the rapid expansion of new knowledge within agronomic sciences. The time lag in publication of texts results in distinct gaps between research information and classroom applications. There is great need for close cooperation among the several educational and research divisions of the societies in preparation of instructional materials.Methodology for attaining improved‐updated instruction are presented for the new comprehensive soil classification system, improved techniques for soil and plant nutrient: deficiency diagnosis and soil pesticide interactions.