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Addressing and limiting burnout and its significant impacts on emergency physicians is an important and ongoing challenge, requiring much more than a focus on individual resilience. This is a key understanding which guides and informs the Australasian College for Emergency Medicine's holistic approach and advocacy efforts when it comes to the wellness of emergency physicians and the health systems in which they operate.

BK polyomavirus (BKPyV) is a ubiquitous pathogen in the human population that is asymptomatic in healthy individuals, but can be life-threatening in those undergoing kidney transplant. To-date, no vaccines or anti-viral therapies are available to treat human BKPyV infections. New therapeutic strategies are urgently required. In this study, using a rational pharmacological screening regimen of known ion channel modulating compounds, we show that BKPyV requires cystic fibrosis transmembrane conductance regulator (CFTR) activity to infect primary renal proximal tubular epithelial cells. Disrupting CFTR function through treatment with the clinically available drug glibenclamide, the CFTR inhibitor CFTR172, or CFTR-silencing, all reduced BKPyV infection. Specifically, time of addition assays and the assessment of the exposure of VP2/VP3 minor capsid proteins indicated a role for CFTR during BKPyV transport to the endoplasmic reticulum, an essential step during the early stages of BKPyV infection. We thus establish CFTR as an important host-factor in the BKPyV life cycle and reveal CFTR modulators as potential anti-BKPyV therapies., Highlights • BK polyomavirus (BKPyV) is life-threatening in those undergoing kidney transplant. • BKPyV requires CFTR to infect primary kidney cells. • Disrupting CFTR function pharmacologically reduces BKPyV infection. • CFTR is required during BKPyV transport to the endoplasmic reticulum.

Despite the availability of prophylactic vaccines the burden of human papillomavirus (HPV) associated malignancy remains high and there is a need to develop additional therapeutic strategies to complement vaccination. We have previously shown that the poorly characterised E5 oncoprotein forms a virus-coded ion channel or viroporin that was sensitive to the amantadine derivative rimantadine. We now demonstrate that alkylated imino sugars, which have antiviral activity against a number of viruses, inhibit E5 channel activity in vitro. Using molecular modelling we predict that imino sugars intercalate between E5 protomers to prevent channel oligomerisation. We explored the ability of these viroporin inhibitors to block E5-mediated activation of mitogenic signalling in keratinocytes. Treatment with either rimantadine or imino sugars prevented ERK-MAPK phosphorylation and reduced cyclin B1 expression in cells expressing E5 from a number of high-risk HPV types. Moreover, viroporin inhibitors also reduced ERK-MAPK activation and cyclin B1 expression in differentiating primary human keratinocytes containing high-risk HPV18. These observations provide evidence of a key role for E5 viroporin function during the HPV life cycle. Viroporin inhibitors could be utilised for stratified treatment of HPV associated tumours prior to virus integration, or as true antiviral therapies to eliminate virus prior to malignant transformation., Highlights • Imino sugars inhibit the viroporin activity of the E5 oncoprotein. • Imino sugars likely interact at E5 protomer interfaces within a channel to prevent oligomerisation. • Imino sugars and adamantanes block mitogenic signalling mediated by E5 from a range of high-risk HPV types. • Viroporin inhibitors reduce mitogenic signalling in differentiating primary keratinocytes containing high-risk HPV18.

Persistent infection with high-risk human papillomavirus (HPV) is the underlying cause of ~5% of all human cancers, including the majority of cervical carcinomas and many other ano-genital and oral cancers. A major challenge remains to identify key host targets of HPV and to reveal how they contribute to virus-mediated malignancy. The HPV E6 oncoprotein aberrantly activates the signal transducer and activator of transcription 3 (STAT3) transcription factor and this is achieved by a virus-driven increase in the levels of the pro-inflammatory cytokine interleukin-6 (IL-6) in HPV positive cervical cancers cells. Crucially, STAT3 activity is essential for the proliferation and survival of cervical cancer cells, suggesting that targeting STAT3 may have therapeutic potential. Unfortunately, the development of direct STAT3 inhibitors has been problematic in the clinic due to toxicity issues identified in early stage trials. To overcome this issue, we focused on the protein Janus kinase 2 (JAK2), which phosphorylates STAT3 and is essential for STAT3 activation. Here, we demonstrate that inhibiting JAK2 reduces cell proliferation and induces apoptosis in HPV transformed cervical cancer cells. We further establish that this is due to inhibition of phosphorylation of the JAK2 substrates STAT3 and STAT5. Finally, we demonstrate that the clinically available JAK2 inhibitor Ruxolitinib synergises with cisplatin in inducing apoptosis, highlighting JAK2 as a promising therapeutic target in HPV-driven cancers.

This essay examines how underlying differing theologies of authority informed mid nineteenth-century Calvinist polemics and ultimately the fragmentation of the New Divinity in New England. Focusing on the polemical career of Nathaniel William Taylor, these differences were evident in earlier Unitarian controversies yet emerged as Taylor and his New Haven allies incrementally departed from historic confessional language.

Chemistry students are often introduced to the concept of atomic orbitals with a representation of a one-dimensional standing wave. The classic example is the harmonic frequencies which produce standing waves on a guitar string; a concept which is easily replicated in class with a length of rope. From here, students are typically exposed to a more realistic three-dimensional model, which can often be difficult to visualize. Extrapolation from a two-dimensional model, such as the vibrational modes of a drumhead, can be used to convey the standing wave concept to students more easily. We have opted to use Chladni plates which may be tuned to give a two-dimensional standing wave which serves as a cross-sectional representation of atomic orbitals. The demonstration, intended for first year chemistry students, facilitates the examination of nodal and anti-nodal regions of a Chladni figure which students can then connect to the concept of quantum mechanical parameters and their relationship to atomic orbital shape.

A fundamental aspect of surgical planning in liver resections is the identification of key vessel tributaries to preserve healthy liver tissue while fully resecting the tumor(s). Current surgical planning relies primarily on the surgeon's ability to mentally reconstruct 2D computed tomography/magnetic resonance (CT/MR) images into 3D and plan resection margins. This creates significant cognitive load, especially for trainees, as it relies on image interpretation, anatomical and surgical knowledge, experience, and spatial sense. The purpose of this study is to determine if 3D reconstruction of preoperative CT/MR images will assist resident-level trainees in making appropriate operative plans for liver resection surgery.Ten preoperative patient CT/MR images were selected. Images were case-matched, 5 to 2D planning and 5 to 3D planning. Images from the 3D group were segmented to create interactive digital models that the resident can manipulate to view the tumor(s) in relation to landmark hepatic structures. Residents were asked to evaluate the images and devise a surgical resection plan for each image. The resident alternated between 2D and 3D planning, in a randomly generated order. The primary outcome was the accuracy of resident's plan compared to expert opinion. Time to devise each surgical plan was the secondary outcome. Residents completed a prestudy and poststudy questionnaire regarding their experience with liver surgery and the 3D planning software.Senior level surgical residents from the Queen's University General Surgery residency program were recruited to participate.A total of 14 residents participated in the study. The median correct response rate was 2 of 5 (40%; range: 0-4) for the 2D group, and 3 of 5 (60%; range: 1-5) for the 3D group (p0.01). The average time to complete each plan was 156 ± 107 seconds for the 2D group, and 84 ± 73 seconds for the 3D group (p0.01). A total 13 of 14 residents found the 3D model easier to use than the 2D. Most residents noticed a difference between the 2 modalities and found that the 3D model improved their confidence with the surgical plan proposed.The results of this study show that 3D reconstruction for liver surgery planning increases accuracy of resident surgical planning and decreases amount of time required. 3D reconstruction would be a useful model for improving trainee understanding of liver anatomy and surgical resection, and would serve as an adjunct to current 2D planning methods. This has the potential to be developed into a module for teaching liver surgery in a competency-based medical curriculum.

Background A number of prominent surgical trials and clinical guidelines regard length of hospital stay and rates of daycase surgery as being of upmost importance following cholecystectomy. However, it is unclear whether these outcomes also matter to patients. This study aimed to identify the factors patients regard as most important when admitted with acute gallstone pathology. Methods A 41-item survey was produced by combining outcomes assessed in recent clinical trials with results from a preliminary patient questionnaire. This was then given out prospectively to patients presenting with acute gallstone pathology, prior to their cholecystectomy. Patients were asked to read an information sheet about laparoscopic cholecystectomy and then complete the survey, scoring each item out of 100 in terms of importance to them. Results Fifty-six patients completed the survey (43 females; median age 51 years). Diagnoses were: cholecystitis (28 patients), biliary colic (13), pancreatitis (10), common bile duct stones (3) and cholangitis (2). The top-scoring survey item was “long-term quality of life after surgery”, with a median value of 97 out of 100. Other high-scoring items included “cleanliness of the ward environment” and “pain control after surgery” (both 96). The lowest-scoring item was “being treated as a daycase” (54). Conclusion Patients with acute gallstone pathology view long-term quality of life after surgery as the most important factor and daycase surgery as the least important. These results should be considered when planning future surgical trials and clinical guidelines. Electronic supplementary material The online version of this article (doi:10.1007/s00268-016-3854-x) contains supplementary material, which is available to authorized users.

Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase-labelling approach to generate site-specifically N-terminally modified variants of both the A2-B5 heterohexamer and B5 pentamer forms of the toxoid. Both forms of the toxoid are endocytosed by GM1-positive mammalian cells, and while the heterohexameric toxoid was principally localized in the ER, the B5 pentamer showed an unexpected localization in the medial/trans Golgi. This study suggests a future role for specifically-labelled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labelling of lipid-rafts in fixed cells.

Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase-labelling approach to generate site-specifically N-terminally modified variants of both the A2-B5 heterohexamer and B5 pentamer forms of the toxoid. Both forms of the toxoid are endocytosed by GM1-positive mammalian cells, and while the heterohexameric toxoid was principally localized in the ER, the B5 pentamer showed an unexpected localization in the medial/trans Golgi. This study suggests a future role for specifically-labelled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labelling of lipid-rafts in fixed cells.

The capability to predict fatigue damage continues to be critical for determining the operational life and inspection intervals of connectors and components used in offshore applications. Subsea well intervention systems are subjected to wave induced cyclic bending moments and understanding the fatigue performance of equipment is essential for determining safe operating envelopes.In this paper, a validated fatigue analysis methodology is presented for non-preloaded connectors that are used within subsea well intervention systems. The fatigue analysis methodology addresses limitations in current standards when calculating the fatigue capacities of non-preloaded connectors with different interacting component materials (i.e. low alloy steel and nickel based alloys). The methodology considers the effect on the fatigue life of both non-axisymmetric geometry/loading, FAT loading, as well as the interaction of different connector materials, capturing any potential change in hot spot locations.Three different non-preloaded connections (i.e. consisting of threaded and load shoulder connectors) were analysed using 3-D finite element analysis models, where ΔM-N curves and the associated crack initiation locations were calculated for each connector. Full-scale resonance fatigue tests were successfully performed on the three different connector types, validating the ΔM-N curves calculated using the fatigue analysis methodology. Fatigue failure (i.e. through-wall crack) was achieved in all tests between 100,000 and 5,000,000 cycles matching the predicted crack initiation location from the analysis for each connection. The validated methodology provides accurate calculation of the fatigue life and correct identification of hot spot locations. Using the validated approach described in this paper within the design process can lead to significant improvements in future designs of connectors, enabling safer operational limits and extending the service life of subsea systems.

This paper represents an expert view from Europe of future emerging technologies within the wind energy sector considering their potential, challenges, applications and technology readiness and how they might evolve in the coming years. These technologies were identified as originating primarily from the academic sector, some start-up companies and a few larger industrial entities. The following areas were considered: airborne wind energy, offshore floating concepts, smart rotors, wind-induced energy harvesting devices, blade tip-mounted rotors, unconventional power transmission systems, multi-rotor turbines, alternative support structures, modular high voltage direct current generators, innovative blade manufacturing techniques, diffuser-augmented turbines and small turbine technologies. The future role of advanced multiscale modelling and data availability is also considered. This expert review has highlighted that more research will be required to realise many of these emerging technologies. However, there is a need to identify synergies between fundamental and industrial research by correctly targeting public and private funding in these emerging technology areas as industrial development may outpace more fundamental research faster than anticipated. © 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).

Persistent human papillomavirus (HPV) infection is the leading cause of cervical cancer. Although the fundamental link between HPV infection and oncogenesis is established, the specific mechanisms of virus-mediated transformation remain poorly understood. We previously demonstrated that the HPV encoded E6 protein increases the activity of the proto-oncogenic transcription factor STAT3 in primary human keratinocytes; however, the molecular basis for STAT3 activation in cervical cancer remains unclear. Here, we show that STAT3 phosphorylation in HPV positive cervical cancer cells is mediated primarily via autocrine activation by the pro-inflammatory cytokine Interleukin 6 (IL-6). Antibody-mediated blockade of IL-6 signalling in HPV positive cells inhibits STAT3 phosphorylation, whereas both recombinant IL-6 and conditioned media from HPV positive cells leads to increased STAT3 phosphorylation within HPV negative cervical cancer cells. Interestingly, we demonstrate that non-conventional activation of the transcription factor NFκB, involving the protein kinase Akt, is required for IL-6 production and subsequent STAT3 activation. Our data provides new insights into the molecular re-wiring of cancer cells by HPV E6. We reveal that activation of an IL-6 signalling axis drives the autocrine and paracrine phosphorylation of STAT3 within HPV positive cervical cancers cells. Greater understanding of this pathway provides a potential opportunity for the use of existing clinically approved drugs for the treatment of HPV-mediated cervical cancer. Author Summary Persistent infection with HPV is the predominant cause of anogenital and oral cancers. Transformation requires the re-wiring of signalling pathways in infected cells by virus encoded oncoproteins. At this point a comprehensive understanding of the full range of host pathways necessary for HPV-mediated carcinogenesis is still lacking. In this study we describe a signalling circuit resulting in the aberrant production of the IL-6 cytokine. Mediated by the HPV E6 oncoprotein, it requires activation of the NFκB transcription factor. The autocrine and paracrine actions of IL-6 are essential for STAT3 activation in HPV-positive cervical cancers. This study provides molecular insights into the mechanisms by which a virus encoded oncoprotein activates an oncogenic pathway, and illuminates potential targets for therapeutic intervention.

Streamflow variability and trends in Australia were investigated for 222 high-quality stream gauging stations having 30 years or more continuous unregulated streamflow records. Trend analysis identified seasonal, inter-annual and decadal variability, long-term monotonic trends and step changes in streamflow. Trends were determined for annual total flow, baseflow, seasonal flows, daily maximum flow and three quantiles of daily flow. A distinct pattern of spatial and temporal variation in streamflow was evident across different hydroclimatic regions in Australia. Most of the stations in southeastern Australia spread across New South Wales and Victoria showed a significant decreasing trend in annual streamflow, while increasing trends were retained within the northern part of the continent. No strong evidence of significant trend was observed for stations in the central region of Australia and northern Queensland. The findings from step change analysis demonstrated evidence of changes in hydrologic responses consistent with observed changes in climate over the past decades. For example, in the Murray–Darling Basin, 51 out of 75 stations were identified with step changes of significant reduction in annual streamflow during the middle to late 1990s, when relatively dry years were recorded across the area. Overall, the hydrologic reference stations (HRSs) serve as critically important gauges for streamflow monitoring and changes in long-term water availability inferred from observed datasets. A wealth of freely downloadable hydrologic data is provided at the HRS web portal including annual, seasonal, monthly and daily streamflow data, as well as trend analysis products and relevant site information.

This article opens up the 1914 case of smugglers Hira Naran and Dulabh Vira, to consider an illicit trade in gold between the eastern ports of southern Africa, east Africa and southern Asia. From a newly compiled database of two and a half thousand known gold dealers active in southern Africa between 1906 and 1926, it is evident that illicit gold dealing was a significant, if hidden, aspect of the region’s towns and ports. Gold was informally ‘exported’ from Johannesburg by an amorphous, diverse group of runners to Durban and Lourenco Marques. At the ports, Gujarati firms, increasingly marginalised by racial legislation, took a leading role in transmitting gold, or sonū, eastwards across the Indian Ocean. The article shows, rather than discusses in the abstract, how a transnational method can reveal hitherto obscure but important themes in southern African history. The article argues that a half-millennium-old indigenous gold trade between south-east Africa and India, in apparent decline by the 1870s, was...

α-helix mediated protein-protein interactions (PPIs) represent important targets for small molecule inhibition, however generic approaches to inhibitor design are in their infancy and would benefit from QSAR analyses to rationalize the non-covalent basis of molecular recognition by designed ligands. In this manuscript using a helix mimetic based on an oligoamide scaffold we utilize the power of a modular synthesis to access compounds which can readily be used to understand the non-covalent determinants of hDM2 recognition by this series of cell active p53/hDM2 inhibitors.

Background: The history of reading, writing, and the dissemination of technology is one of epochal change, and each transition – indeed the history of the book – is marked by hybridity. In the mature years of print, publishers, librarians, and scholars had clearly defined and segregated roles. In the digital realm, the boundaries have broken down. Just now we have hybridity of form and of roles in the implementation of new reading environments.Analysis: This article provides: 1) an overview of e-reading environments; 2) a survey of the Dynamic Table of Contexts interface; and 3) a report on the hybrid production process of a particular online text, Regenerations.Conclusion and implications: Regenerations could only have emerged from a collaboration among a digital infrastructure project, research project, university press, and digital humanities tool suite.

ObjectiveThis study aims to assess treatment compliance among women undergoing definitive chemoradiation with weekly cisplatin for cervical cancer within a safety net health system and to quantify the impact of chemotherapy compliance on outcomes.Materials and MethodsAll women who were treated for International Federation of Gynecology and Obstetrics stage IB2 to stage IVA cervical cancer between April 2008 and May 2014 were identified. Treatment delays were attributed to toxicity, comorbid conditions, or system issues, or categorized as patient-initiated. Disease-free survival and overall survival of women who received fewer than 6 versus 6 or more doses of weekly cisplatin 40 mg/m2 were compared using Kaplan-Meier analyses.ResultsOne hundred nineteen women (mean [SD] age, 48.5 [11.8] years) were identified. Most women (n = 112; 94.1%) completed definitive radiotherapy, requiring a mean (SD) of 56.5 (20.1) days. Sixty-four women (57.1%) completed definitive radiotherapy in 56 days or less. Only 44 women (36.4%) received 6 or more cycles of cisplatin. Of 122 delayed cycles, reasons for delay were as follows: grade 2 or higher toxicity (n = 70; 57.4%), medical comorbidity (n = 12; 9.8%), system issues (n = 9; 7.4%), and patient-initiated (n = 14; 11.5%). Multiple issues complicated treatment for 3 doses (2.5%). Reasons for delay were not documented in 14 doses (11.5%). Among patients who received 6 or more cycles, disease-free survival improved by 17.4 months (mean [SD], 61.1 [3.7] vs 43.7 [4.3] months, P = 0.002) and overall survival improved by 8.6 months (mean [SD], 68.7 [2.3] vs 60.1 [3.7] months, P = 0.011).ConclusionsHigher rates of toxicity and psychosocial barriers to chemotherapy compliance adversely impact survival among women who seek care in low-resource settings. In our population, administration of all 6 cycles of cisplatin was necessary for optimal survival benefit. Future efforts to improve cervical cancer outcomes should address preventable reasons for treatment delays among underinsured or uninsured individuals.

Since the 1960s, a single class of agent has been licensed targeting virus-encoded ion channels, or “viroporins”, contrasting the success of channel blocking drugs in other areas of medicine. Although resistance arose to these prototypic adamantane inhibitors of the influenza A virus (IAV) M2 proton channel, a growing number of clinically and economically important viruses are now recognised to encode essential viroporins providing potential targets for modern drug discovery.We describe the first rationally designed viroporin inhibitor with a comprehensive structure-activity relationship (SAR). This step-change in understanding not only revealed a second biological function for the p7 viroporin from hepatitis C virus (HCV) during virus entry, but also enabled the synthesis of a labelled tool compound that retained biological activity. Hence, p7 inhibitors (p7i) represent a unique class of HCV antiviral targeting both the spread and establishment of infection, as well as a precedent for future viroporin-targeted drug discovery.

Human papillomaviruses (HPV) are a major cause of malignancy worldwide They are the aetiological agent of almost all cervical cancers and an increasing number of head and neck carcinomas. Deregulation of the Hippo pathway component YAP1 has recently been demonstrated to play a role in HPV-mediated cervical cancer, but whether other components of this pathway are implicated in the pathogenesis of this disease remains poorly understood.The expression level and activation status of critical Hippo pathway components were analysed across multiple cytology samples from patients with cervical disease, as well as HPV positive (HPV+) and HPV negative (HPV-) cervical cancer cell lines using real time qPCR, western blot and immunohistochemistry. In parallel, we assessed the effects of MST1 and MST2 overexpression upon cervical cancer cell proliferation, migration and invasion. Finally, we interrogated the consequences of interrupted MST1 and MST2 function using a targeted small molecule inhibitor in tandem with kinase inactive MST mutants. Our analysis found that expression of the Ste20 kinase MST1 was decreased within both HPV+ primary patient samples and cervical cancer cell lines. This effect was mediated by the virus-coded oncoproteins E6 and E7, which impair MST1 transcription. Reintroduction of MST1, or its paralogue MST2, into HPV positive cervical cancer cells re-activated the Hippo pathway, leading to a reduction in cell proliferation, migration and invasion. Finally, using a small molecule inhibitor of MST1/2 or kinase inactive mutants of either protein, we demonstrated that this effect required the kinase function of MST1/2. Our results reveal that HPV down regulates MST1 expression to inactivate the Hippo pathway and so drive cells towards transformation.

BK polyomavirus (BKPyV; hereafter referred to as BK) causes a lifelong chronic infection and is associated with debilitating disease in kidney transplant recipients. Despite its importance, aspects of the virus life cycle remain poorly understood. In addition to the structural proteins, the late region of the BK genome encodes for an auxiliary protein called agnoprotein. Studies on other polyomavirus agnoproteins have suggested that the protein may contribute to virion infectivity. Here, we demonstrate an essential role for agnoprotein in BK virus release. Viruses lacking agnoprotein fail to release from host cells and do not propagate to wild-type levels. Despite this, agnoprotein is not essential for virion infectivity or morphogenesis. Instead, agnoprotein expression correlates with nuclear egress of BK virions. We demonstrate that the agnoprotein binding partner α-soluble N-ethylmaleimide sensitive fusion (NSF) attachment protein (α-SNAP) is necessary for BK virion release, and siRNA knockdown of α-SNAP prevents nuclear release of wild-type BK virions. These data highlight a novel role for agnoprotein and begin to reveal the mechanism by which polyomaviruses leave an infected cell.

Summary BK polyomavirus (BKV) causes polyomavirus-associated nephropathy and hemorrhagic cystitis in immunosuppressed patients. These are diseases for which we currently have limited treatment options, but potential therapies could include pre-transplant vaccination with a multivalent BKV vaccine or therapeutics which inhibit capsid assembly or block attachment and entry into target cells. A useful tool in such efforts would be a high-resolution structure of the infectious BKV virion and how this interacts with its full repertoire of cellular receptors. We present the 3.4-Å cryoelectron microscopy structure of native, infectious BKV in complex with the receptor fragment of GT1b ganglioside. We also present structural evidence that BKV can utilize glycosaminoglycans as attachment receptors. This work highlights features that underpin capsid stability and provides a platform for rational design and development of urgently needed pharmacological interventions for BKV-associated diseases., Graphical Abstract, Highlights • Present the cryo-EM structure of native, infectious BKV virion at 3.4 Å resolution • Reveal interpentamer interactions that mediate capsid assembly • Determine the interaction of BKV with a receptor fragment of GT1b ganglioside • Identify possible sites for glycosaminoglycan binding on the virion surface, BK polyomavirus causes several debilitating diseases in the immunosuppressed with no effective treatments currently available. Hurdiss et al. present the high-resolution structure of BK and complexes with distinct cellular receptors. This work provides a platform for rational design and development of urgently needed targeted pharmacological interventions for BK-associated diseases.

Given the contradictory nature of the literature regarding the role of human papillomaviruses and polyomaviruses in the pathogenesis of urothelial bladder cancer (UBC), we sought to investigate the frequency of their involvement in a large cohort of primary UBCs. DNA was extracted from 689 fresh-frozen UBC tissues and screened for the presence of high-risk human papillomavirus (HPV) types 16 and 18 and BKV/JCV genomic DNA by qPCR. In positive cases, viral identity was confirmed by Sanger sequencing and viral gene expression was analysed by RT-PCR or immunohistochemistry. All 689 UBCs were negative for HPV18. One UBC from a female patient with areas of squamous differentiation was positive for HPV16. The qPCR data indicated variable levels of polyomavirus in 49 UBCs. In the UBCs with low Cts we were able to confirm that 23 were BKV and 6 were JCV by Sanger sequencing. Polyomavirus large T antigen expression was low but detectable in 70% of the sequencing-confirmed polyomavirus positive samples. Thus, in United Kingdom patients, the presence of HPV DNA sequences is extremely rare in UBC (

To assess the effectiveness of prostate artery embolization (PAE) in the control of haematuria and in patients with benign prostatic hyperplasia (BPH) and normal upper urinary tracts. Twelve consecutive patients with haematuria were included in the prospective study. All patients had prior imaging and cystoscopy to exclude other causes of haematuria. Patients prostate arteries were embolized with particles (200–500 μm), and they were followed up at 3, 12 and 18 months following the procedure. QOL questionnaires, IPSS, IIEF and clinical review were all employed to assess the success of the treatment. To allow useful comparison, patients were age- and prostate volume-matched and compared to patients treated with PAE for BPH without haematuria. All 12/12 cases were technically successful with bilateral PAE being performed. All cases of haematuria resolved by the 3-month follow-up (100%). There was one case of recurrence during the 12-month follow-up (overall clinical success at 18 months 92%). This was due to over anticoagulation and ceased once corrected. There was a reduction in lower urinary tract symptoms noted by improvements in QOL indices, IPSS and IIEF. There was continued success even if the patient was subsequently anticoagulated. There was no associated sexual dysfunction. There was more prostatic arterial branching and volume of embolic required to achieve stasis in BPH and haematuria than in BPH alone (p

From increased complexity in compliance to intensified competition, banks face numerous challenges. None is more pressing than achieving excellence in the client experience. Bankers, says Andrew MacDonald, need to revisit a simple but far reaching question: why should anyone bank with you?

Merkel cell carcinoma (MCC) is an aggressive skin cancer of neuroendocrine origin with a high propensity for recurrence and metastasis. Merkel cell polyomavirus (MCPyV) causes the majority of MCC cases due to the expression of the MCPyV small and large tumor antigens (ST and LT, respectively). Although a number of molecular mechanisms have been attributed to MCPyV tumor antigen-mediated cellular transformation or replication, to date, no studies have investigated any potential link between MCPyV T antigen expression and the highly metastatic nature of MCC. Here we use a quantitative proteomic approach to show that MCPyV ST promotes differential expression of cellular proteins implicated in microtubule-associated cytoskeletal organization and dynamics. Intriguingly, we demonstrate that MCPyV ST expression promotes microtubule destabilization, leading to a motile and migratory phenotype. We further highlight the essential role of the microtubule-associated protein stathmin in MCPyV ST-mediated microtubule destabilization and cell motility and implicate the cellular phosphatase catalytic subunit protein phosphatase 4C (PP4C) in the regulation of this process. These findings suggest a possible molecular mechanism for the highly metastatic phenotype associated with MCC. IMPORTANCE Merkel cell polyomavirus (MCPyV) causes the majority of cases of Merkel cell carcinoma (MCC), an aggressive skin cancer with a high metastatic potential. However, the molecular mechanisms leading to virally induced cancer development have yet to be fully elucidated. In particular, no studies have investigated any potential link between the virus and the highly metastatic nature of MCC. We demonstrate that the MCPyV small tumor antigen (ST) promotes the destabilization of the host cell microtubule network, which leads to a more motile and migratory cell phenotype. We further show that MCPyV ST induces this process by regulating the phosphorylation status of the cellular microtubule-associated protein stathmin by its known association with the cellular phosphatase catalytic subunit PP4C. These findings highlight stathmin as a possible biomarker of MCC and as a target for novel antitumoral therapies.

ABSTRACT Mucosal human papillomaviruses express a small, hydrophobic, protein called E5, which plays an important role in the HPV life cycle by delaying normal epithelial cell differentiation while maintaining cell cycle progression. In addition, E5 exhibits transforming abilities in a number of cell culture systems and transgenic mouse models. Lacking any described enzymatic activity, E5 is thought to function by binding to host proteins and modulating their activities. In particular, members of the growth factor receptor family are known targets for subversion. This review article summarizes our latest understanding of this enigmatic oncoprotein, including its role in the HPV life cycle, interactions with host proteins and contribution toward tumorigenesis.

Deregulation of proliferation and differentiation-dependent signalling pathways is a hallmark of human papillomavirus (HPV) infection. Although the manipulation of these pathways by E6 and E7 has been extensively studied, controversies surround the role of the E5 oncoprotein during a productive virus life cycle. By integrating primary keratinocytes harbouring wild type or E5 knockout HPV18 genomes with pharmacological and gain/loss of function models, this study aimed to provide molecular information about the role of E5 in epithelial proliferation and differentiation. We show that E5 contributes to cell cycle progression and unscheduled host DNA synthesis in differentiating keratinocytes. E5 function correlates with increased EGFR activation in differentiating cells and blockade of this pathway impairs differentiation-dependent cell cycle progression of HPV18 containing cells. Our findings provide a functional requirement of enhanced EGFR signalling for suprabasal cellular DNA synthesis during the virus life cycle. They also reveal an unrecognised contribution of E5 towards the impaired keratinocyte differentiation observed during a productive HPV infection. E5 suppresses a signalling axis consisting of the keratinocyte growth factor receptor (KGFR) pathway. Inhibition of this pathway compensates for the loss of E5 in knockout cells and re-instates the delay in differentiation. The negative regulation of KGFR involves suppression by the EGFR pathway. Thus our data reveal an unappreciated role for E5-mediated EGFR signalling in orchestrating the balance between proliferation and differentiation in suprabasal cells.

Cell motility and migration is a complex, multistep, and multicomponent process intrinsic to progression and metastasis. Motility is dependent on the activities of integrin receptors and Rho family GTPases, resulting in the remodeling of the actin cytoskeleton and formation of various motile actin-based protrusions. Merkel cell carcinoma (MCC) is an aggressive skin cancer with a high likelihood of recurrence and metastasis. Merkel cell polyomavirus (MCPyV) is associated with the majority of MCC cases, and MCPyV-induced tumorigenesis largely depends on the expression of the small tumor antigen (ST). Since the discovery of MCPyV, a number of mechanisms have been suggested to account for replication and tumorigenesis, but to date, little is known about potential links between MCPyV T antigen expression and the metastatic nature of MCC. Previously, we described the action of MCPyV ST on the microtubule network and how it impacts cell motility and migration. Here, we demonstrate that MCPyV ST affects the actin cytoskeleton to promote the formation of filopodia through a mechanism involving the catalytic subunit of protein phosphatase 4 (PP4C). We also show that MCPyV ST-induced cell motility is dependent upon the activities of the Rho family GTPases Cdc42 and RhoA. In addition, our results indicate that the MCPyV ST-PP4C interaction results in the dephosphorylation of β1 integrin, likely driving the cell motility pathway. These findings describe a novel mechanism by which a tumor virus induces cell motility, which may ultimately lead to cancer metastasis, and provides opportunities and strategies for targeted interventions for disseminated MCC. IMPORTANCE Merkel cell polyomavirus (MCPyV) is the most recently discovered human tumor virus. It causes the majority of cases of Merkel cell carcinoma (MCC), an aggressive skin cancer. However, the molecular mechanisms implicating MCPyV-encoded proteins in cancer development are yet to be fully elucidated. This study builds upon our previous observations, which demonstrated that the MCPyV ST antigen enhances cell motility, providing a potential link between MCPyV protein expression and the highly metastatic nature of MCC. Here, we show that MCPyV ST remodels the actin cytoskeleton, promoting the formation of filopodia, which is essential for MCPyV ST-induced cell motility, and we also implicate the activity of specific Rho family GTPases, Cdc42 and RhoA, in these processes. Moreover, we describe a novel mechanism for the activation of Rho-GTPases and the cell motility pathway due to the interaction between MCPyV ST and the cellular phosphatase catalytic subunit PP4C, which leads to the specific dephosphorylation of β1 integrin. These findings may therefore provide novel strategies for therapeutic intervention for disseminated MCC.

This paper takes up a more so-called ‘austere’ argument for objectivity derived, for the most part, from P. F. Strawson’s

INTRODUCTION The spleen remains one of the most frequently injured organs following blunt abdominal trauma. In 2012, regional trauma networks were launched across England and Wales with the aim of improving outcomes following trauma. This retrospective cohort study investigated the management and outcomes of blunt splenic injuries before and after the establishment of regional trauma networks. METHODS A dataset was drawn from the Trauma Audit Research Network database of all splenic injuries admitted to English and Welsh hospitals from 1 April 2010 to 31 March 2014. Demographic data, injury severity, treatment modalities and outcomes were collected. Management and outcomes were compared before and after the launch of regional trauma networks. RESULTS There were 1457 blunt splenic injuries: 575 between 2010 and 2012 and 882 in 2012–14. Following the introduction of the regional trauma networks, use of splenic artery embolotherapy increased from 3.5% to 7.6% (P = 0.001) and splenectomy rates decreased from 20% to 14.85% (P = 0.012). Significantly more patients with polytrauma and blunt splenic injury were treated with splenic embolotherapy following 2012 (61.2% vs. 30%, P CONCLUSIONS This study demonstrates a reduction in splenectomy rate and an increased use of splenic artery embolotherapy since the introduction of the regional trauma networks. This may have resulted from improved access to specialist services and reduced practice variation since the establishment of these networks.

Objective This study assessed patterns of superficial reflux in patients with primary chronic venous disease. Methods Retrospective review of all patient venous duplex ultrasonography reports at one institution between 2000 and 2009. Legs with secondary, deep or no superficial reflux were excluded. Results In total, 8654 limbs were scanned; 2559 legs from 2053 patients (mean age 52.3 years) were included for analysis. Great saphenous vein reflux predominated (68%), followed by combined great saphenous vein/small saphenous vein reflux (20%) and small saphenous vein reflux (7%). The majority of legs with competent saphenofemoral junction had below-knee great saphenous vein reflux (53%); incompetent saphenofemoral junction was associated with combined above and below-knee great saphenous vein reflux (72%). Isolated small saphenous vein reflux was associated with saphenopopliteal junction incompetence (61%), although the majority of all small saphenous vein reflux limbs had a competent saphenopopliteal junction (57%). Conclusion Superficial venous reflux does not necessarily originate from a saphenous junction. Large prospective studies with interval duplex ultrasonography are required to unravel the natural history of primary chronic venous disease.

A fifth of worldwide cancer cases have an infectious origin, with viral infection being the foremost. One such cancer is Merkel cell carcinoma (MCC), a rare but aggressive skin malignancy. In 2008, Merkel cell polyomavirus (MCPyV) was discovered as the causative agent of MCC. It is found clonally integrated into the majority of MCC tumours, which require MCPyV oncoproteins to survive. Since its discovery, research has begun to reveal the molecular virology of MCPyV, as well as how it induces tumourigenesis. It is thought to be a common skin commensal, found at low levels in healthy individuals. Upon loss of immunosurveillance, MCPyV reactivates, and a heavy viral load is associated with MCC pathogenesis. Although MCPyV is in many ways similar to classical oncogenic polyomaviruses, such as SV40, subtle differences are beginning to emerge. These unique features highlight the singular position MCPyV has as the only human oncogenic polyomavirus, and open up new avenues for therapies against MCC.

Merkel cell carcinoma (MCC) is a highly aggressive skin cancer with a high metastatic potential. The majority of MCC cases are caused by the Merkel cell polyomavirus (MCPyV), through expression of the virus-encoded tumour antigens. Whilst mechanisms attributing tumour antigen expression to transformation are being uncovered, little is known of the mechanisms by which MCPyV persists in the host. We previously identified the MCPyV small T antigen (tAg) as a novel inhibitor of nuclear factor kappa B (NF-kB) signalling and a modulator of the host anti-viral response. Here we demonstrate that regulation of NF-kB activation involves a previously undocumented interaction between tAg and regulatory sub-unit 1 of protein phosphatase 4 (PP4R1). Formation of a complex with PP4R1 and PP4c is required to bridge MCPyV tAg to the NEMO adaptor protein, allowing deactivation of the NF-kB pathway. Mutations in MCPyV tAg that fail to interact with components of this complex, or siRNA depletion of PP4R1, prevents tAg-mediated inhibition of NF-kB and pro-inflammatory cytokine production. Comparison of tAg binding partners from other human polyomavirus demonstrates that interactions with NEMO and PP4R1 are unique to MCPyV. Collectively, these data identify PP4R1 as a novel target for virus subversion of the host anti-viral response.

To investigate the clinical impact of performing prostate artery embolization (PAE) on patients with adenomatous-dominant benign prostatic hyperplasia (AdBPH). Twelve patients from the ongoing proSTatic aRtery EmbolizAtion for the treatMent of benign prostatic hyperplasia (STREAM) trial were identified as having AdBPH; defined as two or more adenomas within the central gland of ≥1 cm diameter on multi-parametric MRI (MP-MRI). These patients were age-matched with patients from the STREAM cohort, without AdBPH. Patients were followed up with repeat MP-MRI at 3 months and 1 year. International prostate symptom score (IPSS), international index for erectile function (IIEF), and quality of life assessment from the IPSS and EQ-5D-5S questionnaires were recorded pre-PAE and at 6 weeks, 3 months, and 1 year. The mean age of patients was 68 (61–76). All patients had PAE as a day-case procedure. The technical success in the cohort was 23/24 (96%). There was a significant reduction in prostate volume following embolization with a median reduction of 34% (30–55) in the AdBPH group, compared to a mean volume reduction of 22% (9–44) in the non-AdBPH group (p = 0.04). There was a significant reduction in IPSS in the AdBPH group following PAE when compared with the control group [AdBPH median IPSS 8 (3–15) vs. non-AdBPH median IPSS 13 (8–18), p = 0.01]. IPSS QOL scores significantly improved in the AdBPH group (p = 0.007). There was no deterioration in sexual function in either group post-PAE. This is the first time that AdBPH has been identified as being a predictor of clinical success following PAE.

The management of blunt splenic injuries (BSI) has evolved toward strategies that avoid splenectomy. There is growing adoption of interventional radiology (IR) techniques in non-operative management of BSI, with evidence suggesting a corresponding reduction in emergency laparotomy requirements and increased splenic preservation rates. Currently there are no UK national guidelines for the management of blunt splenic injury. This may lead to variations in management, despite the reorganisation of trauma services in England in 2012.A survey was distributed through the British Society of Interventional Radiologists to all UK members aiming to identify availability of IR services in England, radiologists' practice, and attitudes toward management of BSI.116 responses from respondents working in 23 of the 26 Regional Trauma Networks in England were received. 79% provide a single dedicated IR service but over 50% cover more than one hospital within the network. All offer arterial embolisation for BSI. Only 25% follow guidelines. In haemodynamically stable patients, an increasing trend for embolisation was seen as grade of splenic injury increased from 1 to 4 (12.5%-82.14%, p0.01). In unstable patients or those with radiological evidence of bleeding, significantly more respondents offer embolisation for grade 1-3 injuries (p0.01), compared to stable patients. Significantly fewer respondents offer embolisation for grade 5 versus 4 injuries in unstable patients or with evidence of bleeding.Splenic embolisation is offered for a variety of injury grades, providing the patient remains stable. Variation in interventional radiology services remain despite the introduction of regional trauma networks.

O1 IL-15 primes an mTOR-regulated gene-expression program to prolong anti-tumor capacity of human natural killer cells #### Andreas Lundqvist1, Vincent van Hoef1, Xiaonan Zhang1, Erik Wennerberg2, Julie Lorent1, Kristina Witt1, Laia Masvidal Sanz1, Shuo Liang1, Shannon Murray3, Ola Larsson1