Mutation of Vaccinia Virus Gene G2R Causes Suppression of Gene A18R ts Mutants: Implications for Control of Transcription

AbstractThis report provides genetic evidence that two vaccinia virus genes, A18R and G2R, both of which affect the fidelity of viral transcriptionin vivo,interact with each other or act on a common biochemical pathway. Previous experiments with the antipoxviral drug isatin-β-thiosemicarbazone suggest that lethal mutation of gene G2R would compensate for mutations in gene A18R. We therefore tested the hypothesis that gene G2R is an extragenic suppressor of A18R mutations. First, we constructed a recombinant which contains both a G2R deletion mutation and an A18R temperature-sensitive mutation and found that this recombinant was viable. Second, we isolated both cold-sensitive and temperature-insensitive phenotypic revertants of A18R temperature-sensitive mutants and found in both cases that the revertants contained G2R mutations. In the case of the cold-sensitive revertants, we were able to prove that the cold-sensitive phenotype mapped to the G2R gene. Combined with the biochemical data on A18R and G2R, these results imply that the A18R and G2R genes interact with each other either directly or indirectly in a fashion which affects the fidelity of intermediate and late viral transcription

The Vaccinia Virus Bifunctional Gene J3 (Nucleoside-2′-O-)-methyltransferase and Poly(A) Polymerase Stimulatory Factor Is Implicated as a Positive Transcription Elongation Factor by Two Genetic Approaches

AbstractVaccinia virus genes A18 and G2 affect the elongation and termination of postreplicative viral gene transcription in opposite ways. Viruses with mutations in gene A18 produce abnormally long transcripts, indicating that A18 is a negative transcription elongation factor. Viruses containing mutations in gene G2 produce transcripts that are abnormally short, truncated specifically from their 3′ ends, indicating that G2 is a positive transcription elongation factor. Despite the fact that both A18 and G2 are essential genes, A18-G2 double-mutant viruses are viable, presumably because the effects of the mutations are mutually compensatory. In addition, the anti-poxviral drug isatin-β-thiosemicarbazone (IBT) seems to enhance elongation during a vaccinia infection: IBT treatment of a wildtype vaccinia infection induces a phenotype identical to an A18 mutant infection, and G2 mutant viruses are dependent on IBT for growth, presumably because IBT restores the G2 mutant truncated transcripts to a normal length. These observations inspire two independent genetic selections that have now been used to identify an additional vaccinia gene, J3, that regulates postreplicative transcription elongation. In the first selection, a single virus that contains an extragenic suppressor of the A18 temperature-sensitive mutant, Cts23, was isolated. In the second selection, several spontaneous IBT-dependent (IBTd) mutant viruses were isolated and characterized genetically. Marker rescue mapping and DNA sequence analysis show that the extragenic suppressor of Cts23 contains a point mutation in the J3 gene, while each of seven new IBTd mutants contains null mutations in the J3 gene. The J3 protein has previously been identified as a (nucleoside-2′-O-)-methyltransferase and as a processivity subunit for the heterodimeric viral poly(A) polymerase. The nature of the two independent selections used to isolate the J3 mutants strongly suggests that the J3 protein serves as a positive postreplicative transcription elongation factor during a normal virus infection

A randomised, controlled study of outcome and cost effectiveness for RA patients attending nurse-led rheumatology clinics: Study protocol of an ongoing nationwide multi-centre study

Background: The rise in the number of patients with arthritis coupled with understaffing of medical services has seen the deployment of Clinical Nurse Specialists in running nurse-led clinics alongside the rheumatologist clinics. There are no systematic reviews of nurse-led care effectiveness in rheumatoid arthritis. Few published RCTs exist and they have shown positive results for nurse-led care but they have several limitations and there has been no economic assessment of rheumatology nurse-led care in the UK. Objective: This paper outlines the study protocol and methodology currently being used to evaluate the outcomes and cost effectiveness for patients attending rheumatology nurse-led clinics. Design and methods: A multi-centred, pragmatic randomised controlled trial with a non-inferiority design; the null hypothesis being that of 'inferiority' of nurse-led clinics compared to physician-led clinics. The primary outcome is rheumatoid arthritis disease activity (measured by DAS28 score) and secondary outcomes are quality of life, self-efficacy, disability, psychological well-being, satisfaction, pain, fatigue and stiffness. Cost effectiveness will be measured using the EQ-5D, DAS28 and cost profile for each centre. Power calculations: In this trial, a DAS28 change of 0.6 is considered to be the threshold for clinical distinction of 'inferiority'. A sample size of 180 participants (90 per treatment arm) is needed to reject the null hypothesis of 'inferiority', given 90% power. Primary analysis will focus on 2-sided 95% confidence interval evaluation of between-group differences in DAS28 change scores averaged over 4 equidistant follow up time points (13, 26, 39 and 52 weeks). Cost effectiveness will be evaluated assessing the joint parameterisation of costs and effects. Results: The study started in July 2007 and the results are expected after July 2011. Trial registration: The International Standard Randomised Controlled Trial Number ISRCTN29803766. © 2011 Elsevier Ltd

Use of Lysolecithin-Permeabilized Infected-Cell Extracts to Investigate thein VitroBiochemical Phenotypes of Poxvirus ts Mutations Altered in Viral Transcription Activity

AbstractLysolecithin permeabilization of vaccinia virus-infected cells was employed to prepare extracts that support faithful transcription initiationin vitroon plasmids possessing early, intermediate, and late viral gene promoters. Conditions which optimize transcription from each promoter were defined. Thein vitrosystem was used to investigate the multifunctional viral mRNA capping enzyme, which also functions as the viral early gene transcription termination factor (VTF) and a viral intermediate gene transcription initiation factor. A low level of signal-dependent termination of early gene transcription was observedin vitrowhich could be elevated by the addition of pure mRNA capping enzyme. VTF-dependent transcription termination was found to be restricted to templates that possessed an early promoter. This restriction mimics that observedin vivoand demonstrates that transcription termination is limited to RNA polymerase molecules that recognize early rather than intermediate or late gene promoters. Extracts prepared from cells infected at the nonpermissive temperature with a virus containing a ts mutation in gene D12L, which encodes the small subunit of VTF, are incapable of supporting both early gene transcription termination and intermediate gene transcription initiation. Both activities are restored upon addition of the purified wild-type mRNA capping enzyme

Weed Robot

Team Weed Scouts has completed our work on a weed-cutting robot for the Girl Scouts of California’s Central Coast. The final robot build provides a solid foundation that can be built and improved upon by future teams. We have completed the robot base and structure, including the chassis, drivetrain, and robot shell. We also completed manufacturing a weed storage bin and canvas cover for the robot. Additionally, we have built a weed scooper, the mechanism that cuts weeds and transports them into a storage compartment. The electronics and programming for remote control of the robot are also implemented. After some testing, we found that the robot had limited functionality. It was able to drive around with slight power issues but unfortunately, the weed scooper was not able to fully cut and transport the weeds. Despite these obstacles, we have created a semi-operable foundation for future teams to optimize, test, and debug. The next steps include adding a weed shredder and developing autonomous robot functionality along with weed identification. Upon project completion, the weed-cutting robot will help maintain the grounds at Camp Arnaz and serve as a source of engineering inspiration for Girl Scouts and other camp visitors

Women and Climate Change Impacts and Action in Canada: Feminist, Indigenous and Intersectional Perspectives

The report was produced through a collaboration of the Canadian Research Institute for the Advancement of Women and the Alliance for Intergenerational Resilience, with funding from Adapting Canadian Work and Workplaces to Climate Change (ACW) and its predecessor project, Work in a Warming World (W3). The researchers found that women face a double threat from social-economic barriers that leave them bearing the brunt of climate change impacts, while being denied a role in developing policies and programs to mitigate climate change - the example given is employment in renewable energy, where women are underrepresented globally. The report points out that the need for women to be acknowledged as agents of change.Adapting Canadian Work and Workplaces to Respond to Climate Chang

The Strayed Reveller, No. 3

The third issue of The Strayed Revellerhttps://scholarworks.sfasu.edu/reveller/1002/thumbnail.jp

Explaining participation in regional transnational social movement organizations

Since the late 1980s, governments have focused intensely on formalizing political and economic relationships within regions. There has also been a concurrent rise in transnational, regional level organizing among social movement activists globally, suggesting the regionalization of 'global civil society.' However, opportunities for participation in transnational associations vary widely across countries. In this article, we examine the influence of international (both global and regional) institutional contexts, citizen participation in international society, and national level factors on varying levels of participation in regional transnational social movement organizations (TSMOs). We use negative binomial regression to examine relationships among these factors at three time points: 1980, 1990, and 2000. We find that in the early time period, citizen network connections to international society facilitated the formation of and participation in regionally organized TSMOs. Over time, however, regional and global institutional contexts were more predictive of participation in regional TSMOs than were international network ties. Our analysis also uncovered how qualitatively different forms of regionalism translated into significantly different levels of TSMO regionalization. In Europe, where the regional institutional structure is more elaborated than elsewhere in the world, the number of regional TSMOs in which citizens participated greatly outpaced that found elsewhere. Irrespective of international, institutional factors, however, state-level features remained crucial to explaining the development of regional TSMO sectors and the variable levels of participation in them. Citizens in states with restrictions on political rights and civil liberties had significantly lower participation in these organizations in 1990 and 2000. Even so, over time, citizens in states with more ties to global and regional multilateral processes found more ways to overcome this disadvantage and strengthen their participation in regional, transnational civil society. © 2007 SAGE Publications

Designing mini-games as micro-learning resources for professional development in multi-cultural organisations

The need for self-directed learning for professional development drives an increase in the delivery of easy to use {\textquoteleft}just-in-time{\textquoteright} resources that respond to the often-dynamic workplace and work culture. This is especially important in the era of globalisation, when the number of employees, who are culturally diverse, increases each year. Most medium and large companies operate in an international environment, and this is due to the expansion of international enterprises with branches in various countries that requires cooperation with foreign clients, and the employment of foreign nationals in their companies. In order to guarantee the effectiveness of workings in companies, there is a need for continuous education in the aspect of the cultural diversity. This paper explores micro-learning, which focuses on delivering brevity through bite-sized learning units or short-term learning activities. Learning content in this case can take many forms, from text to interactive multimedia. These contents are often created on demand, which can sometimes be less contextualised and pedagogically informed. Based on a case study of the need for training on cultural risks in multi-cultural organisations, this paper focuses on the design of mini-games as playful learning resources for supporting an online learning platform that has been developed as a response to this training need. Fifteen mini-games have been developed to complement eight main topics related to cultural risks and to promote reflection, practice and the self-assessment of knowledge acquired through the platform. The main eight topics represent the risk areas identified that include cultural awareness, understanding different cultures, communication, learning styles, hierarchy, team-working, qualities in the working place, and stereotypes through a survey carried out with personnel (n=154) from multi-cultural organisations across five countries - Cyprus, Italy, Latvia, Poland, and the UK. The discussions include unpacking the mapping of pedagogical and gameful design considerations based on Arnab et al.{\textquoteleft}s (2015) Learning Mechanics-Game Mechanics Mapping (LMGM) model. The paper also discusses the findings from the testing of the online platform across 5 countries including 166 participants (two-step testing). The insights provided will be valuable to researchers, practitioners, designers, and developers of micro-learning resources