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Abstract Background The HIV Rev protein is known to facilitate export of incompletely spliced and unspliced viral transcripts to the cytoplasm, a necessary step in virus life cycle. The Rev-mediated nucleo-cytoplasmic transport of nascent viral transcripts, dependents on interaction of Rev with the RRE RNA structural element present in the target RNAs. The C-terminal variant of dsRNA-binding nuclear protein 90 (NF90ctv) has been shown to markedly attenuate viral replication in stably transduced HIV-1 target cell line. Here we examined a mechanism of interference of viral life cycle involving Rev-NF90ctv interaction. Results Since Rev:RRE complex formations depend on protein:RNA and protein:protein interactions, we investigated whether the expression of NF90ctv might interfere with Rev-mediated export of RRE-containing transcripts. When HeLa cells expressed both NF90ctv and Rev protein, we observed that NF90ctv inhibited the Rev-mediated RNA transport. In particular, three regions of NF90ctv protein are involved in blocking Rev function. Moreover, interaction of NF90ctv with the RRE RNA resulted in the expression of a reporter protein coding sequences linked to the RRE structure. Moreover, Rev influenced the subcellular localization of NF90ctv, and this process is leptomycin B sensitive. Conclusion The dsRNA binding protein, NF90ctv competes with HIV Rev function at two levels, by competitive protein:protein interaction involving Rev binding to specific domains of NF90ctv, as well as by its binding to the RRE-RNA structure. Our results are consistent with a model of Rev-mediated HIV-1 RNA export that envisions Rev-multimerization, a process interrupted by NF90ctv.

In recent years, oncolytic viruses have emerged as promising agents for treating various cancers. An oncolytic virus is a non-pathogenic virus that, due to genetic manipulation, tends to replicate in and cause lysis of cancerous cells while leaving healthy cells unaffected. Among these viruses, vaccinia virus is an attractive platform for use as an oncolytic platform due to its 190 Kb genome with a high capacity for encoding therapeutic payloads. Combining oncolytic VV therapy with other conventional cancer treatments has been shown to be synergistic and more effective than monotherapies. Additionally, OVV can be used as a vector to deliver therapeutic payloads, alone or in combination with other treatments, to increase overall efficacy. Here, we present a comprehensive analysis of preclinical and clinical studies that have evaluated the efficacy of oncolytic vaccinia viruses in cancer immunotherapy. We discuss the outcomes of these studies, including tumor regression rates, overall survival benefits, and long-term responses. Moreover, we provide insights into the challenges and limitations associated with oncolytic vaccinia virus- based therapies, including immune evasion mechanisms, potential toxicities, and the development of resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Mirbahari, Da Silva, Zúñiga, Kooshki Zamani, St-Laurent, Totonchi and Azad.)

Background: Despite proven therapeutic effects in inflammatory conditions, the specific mechanisms of phytochemical therapies are not well understood. The transcriptome effects of Traumeel (Tr14), a multicomponent natural product, and diclofenac, a non-selective cyclooxygenase (COX) inhibitor, were compared in a mouse cutaneous wound healing model to identify both known and novel pathways for the anti-inflammatory effect of plant-derived natural products., Methods: Skin samples from abraded mice were analyzed by single-molecule, amplification-free RNAseq transcript profiling at 7 points between 12 and 192 h after injury. Immediately after injury, the wounds were treated with either diclofenac, Tr14, or placebo control (n = 7 per group/time). RNAseq levels were compared between treatment and control at each time point using a systems biology approach., Results: At early time points (12-36 h), both control and Tr14-treated wounds showed marked increase in the inducible COX2 enzyme mRNA, while diclofenac-treated wounds did not. Tr14, in contrast, modulated lipoxygenase transcripts, especially ALOX12/15, and phospholipases involved in arachidonate metabolism. Notably, Tr14 modulated a group of cell-type specific markers, including the T cell receptor, that could be explained by an overarching effect on the type of cells that were recruited into the wound tissue., Conclusions: Tr14 and diclofenac had very different effects on the COX/LOX synthetic pathway after cutaneous wounding. Tr14 allowed normal autoinduction of COX2 mRNA, but suppressed mRNA levels for key enzymes in the leukotriene synthetic pathway. Tr14 appeared to have a broad 'phytocellular' effect on the wound transcriptome by altering the balance of cell types present in the wound., (© 2021. The Author(s).)

Recombinant forms of the spike protein of SARS-CoV-2 and related viruses have proven difficult to produce with good yields in mammalian cells. Given the panoply of potential COVID-19 diagnostic tools and therapeutic candidates that require purified spike protein and its importance for ongoing SARS-CoV-2 research, we have explored new approaches for spike production and purification. Three transient gene expression methods based on PEI-mediated transfection of CHO or HEK293 cells in suspension culture in chemically-defined media were compared for rapid production of full-length SARS-CoV-2 spike ectodomain. A high-cell-density protocol using DXB11-derived CHO BRI/55E1 cells gave substantially better yields than the other methods. Different forms of the spike ectodomain were expressed, including the wild-type SARS-CoV-2 sequence and a mutated form (to favor expression of the full-length spike ectodomain stabilized in pre-fusion conformation), with and without fusion to putative trimerization domains. An efficient two-step affinity purification method was also developed. Ultimately, we have been able to produce highly homogenous preparations of full-length spike, both monomeric and trimeric, with yields of 100-150 mg/L in the harvested medium. The speed and productivity of this method support further development of CHO-based approaches for recombinant spike protein manufacturing., (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)

Conservation practices during the first decade of the millennium predominantly focused on resisting changes and maintaining historical or current conditions, but ever-increasing impacts from climate change have highlighted the need for transformative action. However, little empirical evidence exists on what kinds of conservation actions aimed specifically at climate change adaptation are being implemented in practice, let alone how transformative these actions are. In response, we propose and trial a novel typology-the R-R-T scale, which improves on existing concepts of Resistance, Resilience, and Transformation-that enables the practical application of contested terms and the empirical assessment of whether and to what extent a shift toward transformative action is occurring. When applying the R-R-T scale to a case study of 104 adaptation projects funded since 2011, we find a trend towards transformation that varies across ecosystems. Our results reveal that perceptions about the acceptance of novel interventions in principle are beginning to be expressed in practice.

Background: A variety of DNA-based methods have been applied to identify genetic markers of attention deficit hyperactivity disorder (ADHD), but the connection to RNA-based gene expression has not been fully exploited., Methods: Using well defined cohorts of discordant, monozygotic twins from the Michigan State University Twin Registry, and case-controlled ADHD cases in adolescents, the present studies utilized advanced single molecule RNA sequencing to identify expressed changes in whole blood RNA in ADHD. Multiple analytical strategies were employed to narrow differentially expressed RNA targets to a small set of potential biomarkers of ADHD., Results: RNA markers common to both the discordant twin study and case-controlled subjects further narrowed the putative targets, some of which had been previously associated with ADHD at the DNA level. The potential role of several differentially expressed genes, including ABCB5, RGS2, GAK, GIT1 and 3 members of the galactose metabolism pathway (GALE, GALT, GALK1) are substantiated by prior associations to ADHD and by established mechanistic connections to molecular pathways relevant to ADHD and behavioral control., Conclusions: The convergence of DNA, RNA, and metabolic data suggests these may be promising targets for diagnostics and therapeutics in ADHD.

Our paper, "The Insignificance of Thresholds in Environmental Impact Assessment: An Illustrative Case Study in Canada" received a critique that challenged us on a number of grounds. Namely, that we defame EIA practitioners, that we advocate EIAs to become a scientific enterprise, that we do not recognize the complexity inherent in EIA, and that EIA undergo an independent assessment by regulators. We respond to all of these points, and argue that conflict of interest is an institutional issue (not one of corrupt practitioners), and that we critique the science that forms the basis of evidence in EIA. Further, we show that the complexity and uncertainty in the critique cannot explain the findings from our paper that all cases of impact threshold exceedance were determined to be not significant in EIA. Finally, we compare the significance determinations in proponent reports to final regulator decisions and determine that they are overwhelmingly identical (93-95%). Regulators are financially independent of proponents, but their decisions on significant are heavily dependent on the information and analysis provided by the proponent reports. As regulators rely on these reports, environmental impact assessments must be based on rigorous and transparent analysis.

Effective governance of public forests depends, in part, on public support for changes in forest management, particularly those responding to changes in socio-ecological conditions driven by climate change. Trust in managing authorities and knowledge about forest management have proven influential in shaping public support for policy across different forest managemen contexts. However, little is known about the relationship between public trust and knowledge as it relates to policy support for emerging management strategies for climate adaptation in forests. We use the example of genomics-based assisted migration (within and outside of natural range) in British Columbia's (BC) forests to examine the relative roles of and interactions between trust in different forestry actors and knowledge of forestry in shaping public support for this new and potentially controversial management alternative. Our results, based on an online survey (n = 1953 BC residents), reveal low public trust in governments and the forest industry combined with low levels of public knowledge about forest management. We find that individuals who are more trusting of decision-makers and other important forestry actors hold higher levels of support for assisted migration. Higher levels of forestry knowledge are linked with support for assisted migration within native range, whereas no knowledge effect is observed for assisted migration outside of native range. We discuss the implications of these observations and provide recommendations to more fully engage with the challenges of low levels of trust and knowledge in this context., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Hedgehog (HH) signaling is involved in many physiological processes, and pathway deregulation can result in a wide range of malignancies. Glioma-associated oncogene 1 (GLI1) is a transcription factor and a terminal effector of the HH cascade. Despite its crucial role in tumorigenesis, our understanding of the GLI1 cellular targets is quite limited. In this study, we identified multiple new GLI1 target genes using a combination of different genomic surveys and then subjected them to in-depth validation in human cancer cell lines. We were able to validate >90% of the new targets, which were enriched in functions involved in neurogenesis and regulation of transcription, in at least one type of follow-up experiment. Strikingly, we found that RNA editing of GLI1 can modulate effects on the targets. Furthermore, one of the top targets, FOXS1, a gene encoding a transcription factor previously implicated in nervous system development, was shown to act in a negative feedback loop limiting the cellular effects of GLI1 in medulloblastoma and rhabdomyosarcoma cells. Moreover, FOXS1 is both highly expressed and positively correlated with GLI1 in medulloblastoma samples of the Sonic HH subgroup, further arguing for the existence of FOXS1/GLI1 interplay in human tumors. Consistently, high FOXS1 expression predicts longer relapse-free survival in breast cancer. Overall, our findings open multiple new avenues in HH signaling pathway research and have potential for translational implications., (© 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)

Environmental assessment is the process that decision-makers rely on to predict, evaluate, and prevent biophysical, social, and economic impacts of potential project developments. The determination of significance in environmental assessment is central to environmental management in many nations. We reviewed ten recent environmental impact assessments from British Columbia, Canada and systematically reviewed and scored significance determination and the approaches used by assessors, the use of thresholds in significance determination, threshold exceedances, and the outcomes. Findings of significant impacts were exceedingly rare and practitioners used a combination of significance determination approaches, most commonly relying upon reasoned argumentation. Quantitative thresholds were rarely employed, with less than 10% of the valued components evaluated using thresholds. Even where quantitative thresholds for significance were exceeded, in every case practitioners used a variety of rationales to demote negative impacts to non-significance. These reasons include combinations of scale (temporal and spatial) of impacts, an already exceeded baseline, model uncertainty and/or substituting less stringent thresholds. Governments and agencies can better protect resources by requiring clear and defensible significance determinations, by making government-defined thresholds legally enforceable and accountable, and by requiring or encouraging significance determination through inclusive and collaborative approaches.

The role of forest management in mitigating climate change is a central concern for the Canadian province of British Columbia. The successful implementation of forest management activities to achieve climate change mitigation in British Columbia will be strongly influenced by public support or opposition. While we now have increasingly clear ideas of the management opportunities associated with forest mitigation and some insight into public support for climate change mitigation in the context of sustainable forest management, very little is known with respect to the levels and basis of public support for potential forest management strategies to mitigate climate change. This paper, by describing the results of a web-based survey, documents levels of public support for the implementation of eight forest carbon mitigation strategies in British Columbia's forest sector, and examines and quantifies the influence of the factors that shape this support. Overall, respondents ascribed a high level of importance to forest carbon mitigation and supported all of the eight proposed strategies, indicating that the British Columbia public is inclined to consider alternative practices in managing forests and wood products to mitigate climate change. That said, we found differences in levels of support for the mitigation strategies. In general, we found greater levels of support for a rehabilitation strategy (e.g. reforestation of unproductive forest land), and to a lesser extent for conservation strategies (e.g. old growth conservation, reduced harvest) over enhanced forest management strategies (e.g. improved harvesting and silvicultural techniques). We also highlighted multiple variables within the British Columbia population that appear to play a role in predicting levels of support for conservation and/or enhanced forest management strategies, including environmental values, risk perception, trust in groups of actors, prioritized objectives of forest management and socio-demographic factors.

In recent years, the provision of economic incentives through carbon financing and carbon offsetting has been central to efforts at forest carbon mitigation. However, notwithstanding their potentially important roles in climate policy, forest carbon offsets face numerous barriers which have limited widespread implementation worldwide. This paper uses the case study of the Canadian province of British Columbia to explore the barriers associated with achieving widespread implementation of forest carbon offsets in the next several decades. Drawing on interviews with experts from government, non-governmental organizations, the private sector and First Nations, six main barriers are identified and discussed: (1) deficiencies of carbon markets, (2) limited economic benefits, (3) uncertain climate effectiveness, (4) negative public opinion, (5) limited and uncertain property rights, and (6) governance issues. While respondents from different sectors agreed on various points, divergence was also observed, notably on the trade-off between generating environmentally sound offsets and promoting cost-effective ways to achieve mitigation. We discuss these differences in the context of the goals and objectives of different actors, and offer insights for understanding the uptake (or not) of carbon offset policies., (Copyright © 2017 Elsevier Ltd. All rights reserved.)