Hereditary retinoblastoma iPSC model reveals aberrant spliceosome function driving bone malignancies.

The RB1 gene is frequently mutated in human cancers but its role in tumorigenesis remains incompletely defined. Using an induced pluripotent stem cell (iPSC) model of hereditary retinoblastoma (RB), we report that the spliceosome is an up-regulated target responding to oncogenic stress in RB1-mutant cells. By investigating transcriptomes and genome occupancies in RB iPSC­derived osteoblasts (OBs), we discover that both E2F3a, which mediates spliceosomal gene expression, and pRB, which antagonizes E2F3a, coregulate more than one-third of spliceosomal genes by cobinding to their promoters or enhancers. Pharmacological inhibition of the spliceosome in RB1-mutant cells leads to global intron retention, decreased cell proliferation, and impaired tumorigenesis. Tumor specimen studies and genome-wide TCGA (The Cancer Genome Atlas) expression profile analyses support the clinical relevance of pRB and E2F3a in modulating spliceosomal gene expression in multiple cancer types including osteosarcoma (OS). High levels of pRB/E2F3a­regulated spliceosomal genes are associated with poor OS patient survival. Collectively, these findings reveal an undiscovered connection between pRB, E2F3a, the spliceosome, and tumorigenesis, pointing to the spliceosomal machinery as a potentially widespread therapeutic vulnerability of pRB-deficient cancers.

The Prevalence of Financial Conflict of Interest Disclosures by Endovascular Specialists on X (Twitter).

PURPOSE: To evaluate conflicts of interest (COIs) among interventional radiologists and related specialties who mention specific devices or companies on the social media (SoMe) platform X, formerly Twitter. MATERIALS AND METHODS: In total, 13,809 posts between October 7, 2021, and December 31, 2021, on X were evaluated. Posts by U.S. interventional radiologists and related specialties who mentioned a specific device or company were identified. A positive COI was defined as receiving a payment from the device manufacturer or company within 36 months prior to posting. The Center for Medicare & Medicaid Services Open Payment database was used to identify financial payments. The prevalence and value of COIs were assessed and compared between posts mentioning a device or company and a paired control group using descriptive statistics and chi-squared tests and independent t tests. RESULTS: Eighty posts containing the mention of 100 specific devices or companies were evaluated. COIs were present in 53% (53/100). When mentioning a specific device or product, 40% interventional radiologists had a COI, compared with 62% neurosurgeons. Physicians who mentioned a specific device or company were 3.7 times more likely to have a positive COI relative to the paired control group (53/100 vs 14/100; P < .001). Of the 31 physicians with a COI, the median physician received $2,270. None of the positive COIs were disclosed. CONCLUSIONS: Physicians posting on SoMe about a specific device or company were more likely to have a financial COI than authors of posts not mentioning a specific device or company. No disclosure of any COI was present in the posts, limiting followers' ability to weigh potential bias.

A meta-analysis of the effect of visiting zoos and aquariums on visitors' conservation knowledge, beliefs, and behavior.

Zoos and aquariums are well placed to connect visitors with the issues facing biodiversity globally and many deliver interventions that seek to influence visitors' beliefs and behaviors with respect to conservation. However, despite primary studies evaluating the effect of such interventions, the overall effect of engaging with zoos and the factors that influence this effect remain unclear. We conducted a systematic review to investigate the effect of zoo-led interventions on knowledge, beliefs (attitudes, intentions, self-efficacy, and social norms), and behavior among zoo visitors. These outcomes were identified using the Theory of Planned Behavior as a theoretical lens. We identified and described the nature of zoo-led interventions in 56 studies and used the behavior change technique (BCT) taxonomy to identify 6 specific BCTs used in interventions to date. Multilevel meta-analyses revealed a small to medium positive effect of engaging with zoo-led interventions on outcomes (d+  = 0.40, 95% confidence interval = 0.28-0.51). Specifically, visitors were more knowledgeable about conservation issues, held more favorable attitudes toward conservation, and reported being more likely to act for the benefit of biodiversity. No evidence of publication bias was present. Effect sizes were, however, heterogeneous and subgroup analyses revealed that the nature of the intervention or type of outcome did not explain this variance. Larger effects were, however, found in studies conducted at a single institution relative to research at multiple institutions and studies that used within-participant designs relative to between-participant designs. Taken together, these findings demonstrate how behavior change frameworks can be used to describe zoo-led interventions and supports the assertion that zoos and aquariums can promote changes in beliefs and behaviors that may help protect biodiversity.

Metaanálisis del efecto de la visita a acuarios y zoológicos sobre el conocimiento, creencias y comportamientos de conservación de los visitantes © 2024 by John Wiley & Sons, Inc. Resumen Los zoológicos y los acuarios están bien posicionados para conectar a los visitantes con temas mundiales de biodiversidad y varios cuentan con intervenciones que buscan influir las creencias y el comportamiento de los visitantes con respecto a la conservación. Sin embargo, con todo y los estudios primarios que evalúan el efecto de dichas intervenciones, aún no está claro el efecto general de participar en los zoológicos y los factores que influyen sobre este efecto. Realizamos una revisión sistemática para investigar el efecto de las intervenciones en los zoológicos sobre el conocimiento, creencias (actitud, intención, autosuficiencia y normas sociales) y comportamiento de sus visitantes. Usamos la teoría del comportamiento planeado como lente teórico para identificar los resultados. Identificamos y describimos la naturaleza de las intervenciones en los zoológicos en 56 estudios y usamos la taxonomía de las técnicas de modificación de conducta (TMC) para identificar seis TMC específicas que se usan hoy en día en dichas intervenciones. Los metaanálisis multinivel revelaron un efecto positivo de pequeño a mediano sobre los resultados causado por la participación en las intervenciones de los zoológicos (d+ = 0.40, 95% CI = 0.28 - 0.51). En específico, los visitantes fueron más conocedores sobre los temas de conservación, tuvieron actitudes más favorables hacia la conservación y reportaron tener mayor probabilidad de actuar en beneficio de la biodiversidad. No hubo evidencias de sesgos en las publicaciones. Sin embargo, el tamaño de los efectos fue heterogéneo y el análisis de los subgrupos reveló que la naturaleza de la intervención o el tipo de resultados no explicaban esta varianza. A pesar de esto, encontramos efectos más grandes en los estudios realizados en una sola institución en relación con la investigación realizada en varias instituciones y los estudios que usaron diseños intraparticipantes en relación a los que usaron diseños interparticipantes. Nuestra revisión demuestra cómo los marcos de modificación conductual pueden usarse para describir las intervenciones en los zoológicos y acuarios y respalda la aseveración de que estas instituciones pueden promover cambios en las creencias y el comportamiento que pueden ayudar a proteger la biodiversidad.

Structural insights into the substrate specificity of the endonuclease activity of the influenza virus cap-snatching mechanism.

The endonuclease activity within the influenza virus cap-snatching process is a proven therapeutic target. The anti-influenza drug baloxavir is highly effective, but is associated with resistance mutations that threaten its clinical efficacy. The endonuclease resides within the N-terminal domain of the PA subunit (PAN) of the influenza RNA dependent RNA polymerase, and we report here complexes of PAN with RNA and DNA oligonucleotides to understand its specificity and the structural basis of baloxavir resistance mutations. The RNA and DNA oligonucleotides bind within the substrate binding groove of PAN in a similar fashion, explaining the ability of the enzyme to cleave both substrates. The individual nucleotides occupy adjacent conserved pockets that flank the two-metal active site. However, the 2' OH of the RNA ribose moieties engage in additional interactions that appear to optimize the binding and cleavage efficiency for the natural substrate. The major baloxavir resistance mutation at position 38 is at the core of the substrate binding site, but structural studies and modeling suggest that it maintains the necessary virus fitness via compensating interactions with RNA. These studies will facilitate the development of new influenza therapeutics that spatially match the substrate and are less likely to elicit resistance mutations.

Halide Chemistry in Tin Perovskite Optoelectronics: Bottlenecks and Opportunities.

Tin halide perovskites (Sn HaPs) are the top lead-free choice for perovskite optoelectronics, but the oxidation of perovskite Sn2+ to Sn4+ remains a key challenge. However, the role of inconspicuous chemical processes remains underexplored. Specifically, the halide component in Sn HaPs (typically iodide) has been shown to play a key role in dictating device performance and stability due to its high reactivity. Here we describe the impact of native halide chemistry on Sn HaPs. Specifically, molecular halogen formation in Sn HaPs and its influence on degradation is reviewed, emphasising the benefits of iodide substitution for improving stability. Next, the ecological impact of halide products of Sn HaP degradation and its mitigation are considered. The development of visible Sn HaP emitters via halide tuning is also summarised. Lastly, halide defect management and interfacial engineering for Sn HaP devices are discussed. These insights will inspire efficient and robust Sn HaP optoelectronics.

Targeting epigenetic modulators using PROTAC degraders: Current status and future perspective.

Epigenetic modulators perform critical functions in gene expression for rapid adaption to external stimuli and are prevalent in all higher-order organisms. The establishment of a link between dysregulation of epigenetic processes and disease pathogenesis, particularly in cancer, has led to much interest in identifying drug targets. This prompted the development of small molecule inhibitors, primarily in haematological malignancies. While there have been epigenetic-targeting drugs to receive FDA approval for the treatment of cancers, many suffer from limited applicability, toxicity and the onset of drug resistance, as our understanding of the biology remains incomplete. The recent advent of genome-wide RNAi and CRISPR screens has shed new light on loss of specific proteins causing vulnerabilities of specific cancer types, highlighting the potential for exploiting synthetic lethality as a therapeutic approach. However, small molecule inhibitors have largely been unable to recapitulate phenotypic effects observed using genome-wide knockdown approaches. This mechanistic disconnect and gap are set to be addressed by targeted protein degradation. Degraders such as PROTACs targeting epigenetic proteins recapitulate CRISPR mediated genetic knockdown at the post-translational level and therefore can better exploit target druggability. Here, we review the current landscape of epigenetic drug discovery, the rationale behind and progress made in the development of PROTAC degraders, and look at future perspectives for the field.

Phosphorene Nanoribbon-Augmented Optoelectronics for Enhanced Hole Extraction.

Phosphorene nanoribbons (PNRs) have been widely predicted to exhibit a range of superlative functional properties; however, because they have only recently been isolated, these properties are yet to be shown to translate to improved performance in any application. PNRs show particular promise for optoelectronics, given their predicted high exciton binding energies, tunable bandgaps, and ultrahigh hole mobilities. Here, we verify the theorized enhanced hole mobility in both solar cells and space-charge-limited-current devices, demonstrating the potential for PNRs improving hole extraction in universal optoelectronic applications. Specifically, PNRs are demonstrated to act as an effective charge-selective interlayer by enhancing hole extraction from polycrystalline methylammonium lead iodide (MAPbI3) perovskite to the poly(triarylamine) semiconductor. Introducing PNRs at the hole-transport/MAPbI3 interface achieves fill factors above 0.83 and efficiencies exceeding 21% for planar p-i-n (inverted) perovskite solar cells (PSCs). Such efficiencies are typically only reported for single-crystalline MAPbI3-based inverted PSCs. Methylammonium-free PSCs also benefit from a PNR interlayer, verifying applicability to architectures incorporating mixed perovskite absorber layers. Device photoluminescence and transient absorption spectroscopy are used to demonstrate that the presence of the PNRs drives more effective carrier extraction. Isolation of the PNRs in space-charge-limited-current hole-only devices improves both hole mobility and conductivity, demonstrating applicability beyond PSCs. This work provides primary experimental evidence that the predicted superlative functional properties of PNRs indeed translate to improved optoelectronic performance.

What's hot and what's not: Making sense of biodiversity 'hotspots'.

Conserving biogeographic regions with especially high biodiversity, known as biodiversity 'hotspots', is intuitive because finite resources can be focussed towards manageable units. Yet, biodiversity, environmental conditions and their relationship are more complex with multidimensional properties. Assessments which ignore this risk failing to detect change, identify its direction or gauge the scale of appropriate intervention. Conflicting concepts which assume assemblages as either sharply delineated communities or loosely collected species have also hampered progress in the way we assess and conserve biodiversity. We focus on the marine benthos where delineating manageable areas for conservation is an attractive prospect because it holds most marine species and constitutes the largest single ecosystem on earth by area. Using two large UK marine benthic faunal datasets, we present a spatially gridded data sampling design to account for survey effects which would otherwise be the principal drivers of diversity estimates. We then assess γ-diversity (regional richness) with diversity partitioned between α (local richness) and ß (dissimilarity), and their change in relation to covariates to test whether defining and conserving biodiversity hotspots is an effective conservation strategy in light of the prevailing forces structuring those assemblages. α-, ß- and γ-diversity hotspots were largely inconsistent with each metric relating uniquely to the covariates, and loosely collected species generally prevailed with relatively few distinct assemblages. Hotspots could therefore be an unreliable means to direct conservation efforts if based on only a component part of diversity. When assessed alongside environmental gradients, α-, ß- and γ-diversity provide a multidimensional but still intuitive perspective of biodiversity change that can direct conservation towards key drivers and the appropriate scale for intervention. Our study also highlights possible temporal declines in species richness over 30 years and thus the need for future integrated monitoring to reveal the causal drivers of biodiversity change.

Long-term Results With CorMatrix Extracellular Matrix Patches After Carotid Endarterectomy.

BACKGROUND: Previous reports of extracellular matrix (ECM) patch use after carotid endarterectomy (CEA) have noted an approximately 10% rate of pseudoaneurysm (PSA) formation. PSA-related rupture of ECM patches has also been described after femoral artery repair. In these studies, different thicknesses (4-ply versus 6-ply) and no standard length of soaking the patch in saline before implantation were used. Herein, we describe our experience with ECM CorMatrix patches in 291 CEAs with 6-ply patches. METHODS: The records of 275 consecutive patients undergoing 291 CEAs with CorMatrix 6-ply patches beginning in November of 2011 and extending until 2015 were reviewed. Only 6-ply patches and a 1 min hydration time in saline were used in all patients. No shunts were used. RESULTS: There were three deaths within the first 30 d secondary to subsequent cardiac surgical procedures. Nine patients experienced a perioperative stroke (3.1%), only one of which occurred secondary to an occluded internal carotid artery. One patient had a transient ischemic attack with a patent endarterectomy site. In follow-up, 11 patients (4.5%) developed severe recurrent stenoses requiring reintervention. Only one patient (0.34%) developed a PSA at 2 years possibly secondary to chronic infection. The median follow-up was 72 mo. CONCLUSIONS: Our experience with 6-ply CorMatrix ECM patches and a brief period of soaking demonstrated that these patches performed well in patients requiring a CEA. Only one PSA was noted.

The spliceosome is a therapeutic vulnerability in MYC-driven cancer.

MYC (also known as c-MYC) overexpression or hyperactivation is one of the most common drivers of human cancer. Despite intensive study, the MYC oncogene remains recalcitrant to therapeutic inhibition. MYC is a transcription factor, and many of its pro-tumorigenic functions have been attributed to its ability to regulate gene expression programs. Notably, oncogenic MYC activation has also been shown to increase total RNA and protein production in many tissue and disease contexts. While such increases in RNA and protein production may endow cancer cells with pro-tumour hallmarks, this increase in synthesis may also generate new or heightened burden on MYC-driven cancer cells to process these macromolecules properly. Here we discover that the spliceosome is a new target of oncogenic stress in MYC-driven cancers. We identify BUD31 as a MYC-synthetic lethal gene in human mammary epithelial cells, and demonstrate that BUD31 is a component of the core spliceosome required for its assembly and catalytic activity. Core spliceosomal factors (such as SF3B1 and U2AF1) associated with BUD31 are also required to tolerate oncogenic MYC. Notably, MYC hyperactivation induces an increase in total precursor messenger RNA synthesis, suggesting an increased burden on the core spliceosome to process pre-mRNA. In contrast to normal cells, partial inhibition of the spliceosome in MYC-hyperactivated cells leads to global intron retention, widespread defects in pre-mRNA maturation, and deregulation of many essential cell processes. Notably, genetic or pharmacological inhibition of the spliceosome in vivo impairs survival, tumorigenicity and metastatic proclivity of MYC-dependent breast cancers. Collectively, these data suggest that oncogenic MYC confers a collateral stress on splicing, and that components of the spliceosome may be therapeutic entry points for aggressive MYC-driven cancers.

Identifying the determinants of emotion regulation choice: a systematic review with meta-analysis.

Day-to-day life is inundated with attempts to control emotions and a wealth of research has examined what strategies people use and how effective these strategies are. However, until more recently, research has often neglected more basic questions such as whether and how people choose to regulate their emotions (i.e. emotion regulation choice). In an effort to identify what we know and what we need to know, we systematically reviewed studies that examined potential determinants of whether and how people choose to regulate their emotions. Eighteen determinants were identified across 219 studies and were categorised as being affective, cognitive, motivational, individual or social-cultural in nature. Where there were sufficient primary studies, meta-analysis was used to quantify the size of the associations between potential determinants and measures of whether and how people choose to regulate their emotions. Based on the findings, we propose that people's decisions about whether and how to regulate their emotions are determined by factors relating to the individual doing the regulating, the emotion that is being regulated, and both the immediate situation and the broader social context in which the regulation is taking place.

Genome-wide RNAi screens in human brain tumor isolates reveal a novel viability requirement for PHF5A.

To identify key regulators of human brain tumor maintenance and initiation, we performed multiple genome-wide RNAi screens in patient-derived glioblastoma multiforme (GBM) stem cells (GSCs). These screens identified the plant homeodomain (PHD)-finger domain protein PHF5A as differentially required for GSC expansion, as compared with untransformed neural stem cells (NSCs) and fibroblasts. Given PHF5A's known involvement in facilitating interactions between the U2 snRNP complex and ATP-dependent helicases, we examined cancer-specific roles in RNA splicing. We found that in GSCs, but not untransformed controls, PHF5A facilitates recognition of exons with unusual C-rich 3' splice sites in thousands of essential genes. PHF5A knockdown in GSCs, but not untransformed NSCs, astrocytes, or fibroblasts, inhibited splicing of these genes, leading to cell cycle arrest and loss of viability. Notably, pharmacologic inhibition of U2 snRNP activity phenocopied PHF5A knockdown in GSCs and also in NSCs or fibroblasts overexpressing MYC. Furthermore, PHF5A inhibition compromised GSC tumor formation in vivo and inhibited growth of established GBM patient-derived xenograft tumors. Our results demonstrate a novel viability requirement for PHF5A to maintain proper exon recognition in brain tumor-initiating cells and may provide new inroads for novel anti-GBM therapeutic strategies.

Inhibition of SF3B1 by molecules targeting the spliceosome results in massive aberrant exon skipping.

The recent identification of compounds that interact with the spliceosome (sudemycins, spliceostatin A, and meayamycin) indicates that these molecules modulate aberrant splicing via SF3B1 inhibition. Through whole transcriptome sequencing, we have demonstrated that treatment of Rh18 cells with sudemycin leads to exon skipping as the predominant aberrant splicing event. This was also observed following reanalysis of published RNA-seq data sets derived from HeLa cells after spliceostatin A exposure. These results are in contrast to previous reports that indicate that intron retention was the major consequence of SF3B1 inhibition. Analysis of the exon junctions up-regulated by these small molecules indicated that these sequences were absent in annotated human genes, suggesting that aberrant splicing events yielded novel RNA transcripts. Interestingly, the length of preferred downstream exons was significantly longer than the skipped exons, although there was no difference between the lengths of introns flanking skipped exons. The reading frame of the aberrantly skipped exons maintained a ratio of 2:1:1, close to that of the cassette exons (3:1:1) present in naturally occurring isoforms, suggesting negative selection by the nonsense-mediated decay (NMD) machinery for out-of-frame transcripts. Accordingly, genes involved in NMD and RNAs encoding proteins involved in the splicing process were enriched in both data sets. Our findings, therefore, further elucidate the mechanisms by which SF3B1 inhibition modulates pre-mRNA splicing.

Coding Variation in ANGPTL4, LPL, and SVEP1 and the Risk of Coronary Disease.

BACKGROUND: The discovery of low-frequency coding variants affecting the risk of coronary artery disease has facilitated the identification of therapeutic targets. METHODS: Through DNA genotyping, we tested 54,003 coding-sequence variants covering 13,715 human genes in up to 72,868 patients with coronary artery disease and 120,770 controls who did not have coronary artery disease. Through DNA sequencing, we studied the effects of loss-of-function mutations in selected genes. RESULTS: We confirmed previously observed significant associations between coronary artery disease and low-frequency missense variants in the genes LPA and PCSK9. We also found significant associations between coronary artery disease and low-frequency missense variants in the genes SVEP1 (p.D2702G; minor-allele frequency, 3.60%; odds ratio for disease, 1.14; P=4.2×10(-10)) and ANGPTL4 (p.E40K; minor-allele frequency, 2.01%; odds ratio, 0.86; P=4.0×10(-8)), which encodes angiopoietin-like 4. Through sequencing of ANGPTL4, we identified 9 carriers of loss-of-function mutations among 6924 patients with myocardial infarction, as compared with 19 carriers among 6834 controls (odds ratio, 0.47; P=0.04); carriers of ANGPTL4 loss-of-function alleles had triglyceride levels that were 35% lower than the levels among persons who did not carry a loss-of-function allele (P=0.003). ANGPTL4 inhibits lipoprotein lipase; we therefore searched for mutations in LPL and identified a loss-of-function variant that was associated with an increased risk of coronary artery disease (p.D36N; minor-allele frequency, 1.9%; odds ratio, 1.13; P=2.0×10(-4)) and a gain-of-function variant that was associated with protection from coronary artery disease (p.S447*; minor-allele frequency, 9.9%; odds ratio, 0.94; P=2.5×10(-7)). CONCLUSIONS: We found that carriers of loss-of-function mutations in ANGPTL4 had triglyceride levels that were lower than those among noncarriers; these mutations were also associated with protection from coronary artery disease. (Funded by the National Institutes of Health and others.).

Meta-Analysis of Genome-Wide Association Studies for Abdominal Aortic Aneurysm Identifies Four New Disease-Specific Risk Loci.

RATIONALE: Abdominal aortic aneurysm (AAA) is a complex disease with both genetic and environmental risk factors. Together, 6 previously identified risk loci only explain a small proportion of the heritability of AAA. OBJECTIVE: To identify additional AAA risk loci using data from all available genome-wide association studies. METHODS AND RESULTS: Through a meta-analysis of 6 genome-wide association study data sets and a validation study totaling 10 204 cases and 107 766 controls, we identified 4 new AAA risk loci: 1q32.3 (SMYD2), 13q12.11 (LINC00540), 20q13.12 (near PCIF1/MMP9/ZNF335), and 21q22.2 (ERG). In various database searches, we observed no new associations between the lead AAA single nucleotide polymorphisms and coronary artery disease, blood pressure, lipids, or diabetes mellitus. Network analyses identified ERG, IL6R, and LDLR as modifiers of MMP9, with a direct interaction between ERG and MMP9. CONCLUSIONS: The 4 new risk loci for AAA seem to be specific for AAA compared with other cardiovascular diseases and related traits suggesting that traditional cardiovascular risk factor management may only have limited value in preventing the progression of aneurysmal disease.

The Relationship Between a Balanced Time Perspective and Self-monitoring of Blood Glucose Among People With Type 1 Diabetes.

Background: Self-monitoring of blood glucose helps people with type 1 diabetes to maintain glycemic control and reduce the risk of complications. However, adherence to blood glucose monitoring is often suboptimal. Purpose: Like many health behaviors, self-monitoring of blood glucose involves exerting effort in the present to achieve future benefits. As such, the present research explored whether individual differences in time perspective-specifically, the extent to which people have a balanced time perspective-are associated with the frequency with which people with type 1 diabetes monitor their blood glucose and, thus, maintain glycemic control. Methods: Adults with type 1 diabetes completed measures of time perspective, feelings associated with monitoring, attitudes toward monitoring, and trait self-control. Objective data regarding the frequency with which participants monitored their blood glucose levels and their long-term glycemic control were extracted from their medical records. Results: Hierarchical regression analyses and tests of indirect effects (N = 129) indicated that having a more balanced time perspective was associated with more frequent monitoring of blood glucose and, as a result, better glycemic control. Further analyses (N = 158) also indicated that there was an indirect relationship between balanced time perspective and monitoring of blood glucose via the feelings that participants associated with monitoring and their subsequent attitudes toward monitoring. Conclusions: These findings point to the importance and relevance of time perspective for understanding health-related behavior and may help to inform interventions designed to promote self-monitoring of blood glucose in people with type 1 diabetes.

Identification and characterization of influenza variants resistant to a viral endonuclease inhibitor.

The influenza endonuclease is an essential subdomain of the viral RNA polymerase. It processes host pre-mRNAs to serve as primers for viral mRNA and is an attractive target for antiinfluenza drug discovery. Compound L-742,001 is a prototypical endonuclease inhibitor, and we found that repeated passaging of influenza virus in the presence of this drug did not lead to the development of resistant mutant strains. Reduced sensitivity to L-742,001 could only be induced by creating point mutations via a random mutagenesis strategy. These mutations mapped to the endonuclease active site where they can directly impact inhibitor binding. Engineered viruses containing the mutations showed resistance to L-742,001 both in vitro and in vivo, with only a modest reduction in fitness. Introduction of the mutations into a second virus also increased its resistance to the inhibitor. Using the isolated wild-type and mutant endonuclease domains, we used kinetics, inhibitor binding and crystallography to characterize how the two most significant mutations elicit resistance to L-742,001. These studies lay the foundation for the development of a new class of influenza therapeutics with reduced potential for the development of clinical endonuclease inhibitor-resistant influenza strains.

Uninformed Reform: The Attempt to Abolish the Hospital Managers' Section 23 Discharge Power Under the Mental Health Act 1983.

Under section 23 of the Mental Health Act 1983 a person can be discharged by the managers of the hospital from compulsory care. The limited evidence indicates that the section 23 power is normally delegated to a specially appointed panel who hold a hearing. Unfortunately, notwithstanding the implications for the liberty, autonomy, and dignity of the compelled person, very little is known about how this process operates. Nonetheless, since 1996 there has been a sustained effort to abolish the power. In view of this, the proposal to reform the 1983 Act contained in the Queen's Speech January 2017, and the subsequent establishment of the Independent Review of the Mental Health Act in October 2017, I critique the claims made in the abolition debate, and establish the conceptual gaps therein. I argue that a much more developed understanding of the power is required before any change is made to the law in this area.

USP39 Deubiquitinase Is Essential for KRAS Oncogene-driven Cancer.

KRAS is the most frequently mutated oncogene in human cancer, but its therapeutic targeting remains challenging. Here, we report a synthetic lethal screen with a library of deubiquitinases and identify USP39, which encodes an essential splicing factor, as a critical gene for the viability of KRAS-dependent cells. We show that splicing fidelity inhibitors decrease preferentially the proliferation rate of KRAS-active cells. Moreover, depletion of DHX38, encoding an USP39-interacting splicing factor, also reduces the viability of these cells. In agreement with these results, USP39 depletion caused a significant reduction in pre-mRNA splicing efficiency, as demonstrated through RNA-seq experiments. Furthermore, we show that USP39 is up-regulated in lung and colon carcinomas and its expression correlates with KRAS levels and poor clinical outcome. Accordingly, our work provides critical information for the development of splicing-directed antitumor treatments and supports the potential of USP39-targeting strategies as the basis of new anticancer therapies.

Increased energy differentially increases richness and abundance of optimal body sizes in deep-sea wood falls.

Theoretical and empirical studies suggest that the total energy available in natural communities influences body size as well as patterns of abundance and diversity. But the precise mechanisms underlying these relationships or how these three ecological properties relate remain elusive. We identify five hypotheses relating energy availability, body size distributions, abundance, and species richness within communities, and we use experimental deep-sea wood fall communities to test their predicted effects both on descriptors describing the species-richness-body-size distribution, and on trends in species richness within size classes over an energy gradient (size-class-richness relationships). Invertebrate communities were taxonomically identified, weighed, and counted from 32 Acacia sp. logs ranging in size from 0.6 to 20.6 kg (corresponding to different levels of energy available), which were deployed at 3,203 m in the Northeast Pacific Ocean for 5 and 7 yr. Trends in both the species-richness-body-size distribution and the size-class-richness distribution with increasing wood fall size provide support for the Increased Packing hypothesis: species richness increases with increasing wood fall size but only in the modal size class. Furthermore, species richness of body size classes reflected the abundance of individuals in that size class. Thus, increases in richness in the modal size class with increasing energy were concordant with increases in abundance within that size class. The results suggest that increases in species richness occurring as energy availability increases may be isolated to specific niches, e.g., the body size classes, especially in communities developing on discrete and energetically isolated resources such as deep sea wood falls.