TRIM7 inhibits enterovirus replication and promotes emergence of a viral variant with increased pathogenicity.

To control viral infection, vertebrates rely on both inducible interferon responses and less well-characterized cell-intrinsic responses composed of "at the ready" antiviral effector proteins. Here, we show that E3 ubiquitin ligase TRIM7 is a cell-intrinsic antiviral effector that restricts multiple human enteroviruses by targeting viral 2BC, a membrane remodeling protein, for ubiquitination and proteasome-dependent degradation. Selective pressure exerted by TRIM7 results in emergence of a TRIM7-resistant coxsackievirus with a single point mutation in the viral 2C ATPase/helicase. In cultured cells, the mutation helps the virus evade TRIM7 but impairs optimal viral replication, and this correlates with a hyperactive and structurally plastic 2C ATPase. Unexpectedly, the TRIM7-resistant virus has a replication advantage in mice and causes lethal pancreatitis. These findings reveal a unique mechanism for targeting enterovirus replication and provide molecular insight into the benefits and trade-offs of viral evolution imposed by a host restriction factor.

Macaques recognize features in synthetic images derived from ventral stream neurons.

Primates can recognize features in virtually all types of images, an ability that still requires a comprehensive computational explanation. One hypothesis is that visual cortex neurons learn patterns from scenes, objects, and textures, and use these patterns to interpolate incoming visual information. We have used machine learning algorithms to instantiate visual patterns stored by neurons-we call these highly activating images prototypes. Prototypes from inferotemporal (IT) neurons often resemble parts of real-world objects, such as monkey faces and body parts, a similarity established via pretrained neural networks [C. R. Ponce et al., Cell 177, 999-1009.e10 (2019)] and naïve human participants [A. Bardon, W. Xiao, C. R. Ponce, M. S. Livingstone, G. Kreiman, Proc. Natl. Acad. Sci. U.S.A. 119, e2118705119 (2022)]. However, it is not known whether monkeys themselves perceive similarities between neuronal prototypes and real-world objects. Here, we investigated whether monkeys reported similarities between prototypes and real-world objects using a two-alternative forced choice task. We trained the animals to saccade to synthetic images of monkeys, and subsequently tested how they classified prototypes synthesized from IT and primary visual cortex (V1). We found monkeys classified IT prototypes as conspecifics more often than they did random generator images and V1 prototypes, and their choices were partially predicted by convolutional neural networks. Further, we confirmed that monkeys could abstract general shape information from images of real-world objects. Finally, we verified these results with human participants. Our results provide further evidence that prototypes from cortical neurons represent interpretable abstractions from the visual world.

Shiftless inhibits flavivirus replication in vitro and is neuroprotective in a mouse model of Zika virus pathogenesis.

Flaviviruses such as Zika virus and West Nile virus have the potential to cause severe neuropathology if they invade the central nervous system. The type I interferon response is well characterized as contributing to control of flavivirus-induced neuropathogenesis. However, the interferon-stimulated gene (ISG) effectors that confer these neuroprotective effects are less well studied. Here, we used an ISG expression screen to identify Shiftless (SHFL, C19orf66) as a potent inhibitor of diverse positive-stranded RNA viruses, including multiple members of the Flaviviridae (Zika, West Nile, dengue, yellow fever, and hepatitis C viruses). In cultured cells, SHFL functions as a viral RNA-binding protein that inhibits viral replication at a step after primary translation of the incoming genome. The murine ortholog, Shfl, is expressed constitutively in multiple tissues, including the central nervous system. In a mouse model of Zika virus infection, Shfl-/- knockout mice exhibit reduced survival, exacerbated neuropathological outcomes, and increased viral replication in the brain and spinal cord. These studies demonstrate that Shfl is an important antiviral effector that contributes to host protection from Zika virus infection and virus-induced neuropathological disease.

Audit and feedback interventions involving pharmacists to influence prescribing behaviour in general practice: a systematic review and meta-analysis.

INTRODUCTION: Pharmacists, as experts in medicines, are increasingly employed in general practices and undertake a range of responsibilities. Audit and feedback (A&F) interventions are effective in achieving behaviour change, including prescribing. The extent of pharmacist involvement in A&F interventions to influence prescribing is unknown. This review aimed to assess the effectiveness of A&F interventions involving pharmacists on prescribing in general practice compared with no A&F/usual care and to describe features of A&F interventions and pharmacist characteristics. METHODS: Electronic databases (MEDLINE, EMBASE, CINAHL, Cochrane Central Register of Controlled Trials, (Social) Science Citation Indexes, ISI Web of Science) were searched (2012, 2019, 2020). Cochrane systematic review methods were applied to trial identification, selection, and risk of bias. Results were summarized descriptively and heterogeneity was assessed. A random-effects meta-analysis was conducted where studies were sufficiently homogenous in design and outcome. RESULTS: Eleven cluster-randomized studies from 9 countries were included. Risk of bias across most domains was low. Interventions focussed on older patients, specific clinical area(s), or specific medications. Meta-analysis of 6 studies showed improved prescribing outcomes (pooled risk ratio: 0.78, 95% confidence interval: 0.64-0.94). Interventions including both verbal and written feedback or computerized decision support for prescribers were more effective. Pharmacists who received study-specific training, provided ongoing support to prescribers or reviewed prescribing for individual patients, contributed to more effective interventions. CONCLUSIONS: A&F interventions involving pharmacists can lead to small improvements in evidence-based prescribing in general practice settings. Future implementation of A&F within general practice should compare different ways of involving pharmacists to determine how to optimize effectiveness.PRISMA-compliant abstract included in Supplementary Material 1.

Trends in COVID-19 cases, deaths, and staffing shortages in US nursing homes by rural and urban status.

The purpose of the study was to examine trends in COVID-19 cases, related deaths, and staffing shortages in nursing homes (NH) by rural and urban status from May 2020 to Feb 2021. Generalized linear mixed models with state-fixed effects were used to estimate the interaction effect of study period and rural/urban status on having at least: one COVID-19 case, one related death, and/or at least one week of staffing shortage using the NH COVID-19 data spanning the 40-week period. The findings revealed shortages in staff, particularly direct care providers, were greatly accelerated in rural NHs as the pandemic wore on over time. Conversely, staffing shortages in urban NHs were relatively stable despite the fluctuating COVID-19 cases over the same time period. The findings highlight the need of identifying effective strategies that prevent rural NHs from encountering staffing deficits in response to long-lasting natural disasters such as the COVID-19 pandemic.

An implanted port-catheter system for repeated hepatic arterial infusion of low-density lipoprotein-docosahexaenoic acid nanoparticles in normal rats: A safety study.

BACKGROUND: In recent years, small animal arterial port-catheter systems have been implemented in rodents with reasonable success. The aim of the current study is to employ the small animal port-catheter system to evaluate the safety of multiple hepatic-artery infusions (HAI) of low-density lipoprotein-docosahexaenoic acid (LDL-DHA) nanoparticles to the rat liver. METHODS: Wistar rats underwent surgical placement of indwelling HAI ports. Repeated administrations of PBS or LDL-DHA nanoparticles were performed through the port at baseline and days 3 and 6. Rats were sacrificed on day 9 at which point blood and various organs were collected for histopathology and biochemical analyses. RESULTS: The port-catheter systems were implanted successfully and repeated infusions of PBS or LDL-DHA nanoparticles were tolerated well by all animals over the duration of the study. Measurements of serum liver/renal function tests, glucose and lipid levels did not differ between control and LDL-DHA treated rats. The liver histology was unremarkable in the LDL-DHA treated rats and the expression of hepatic inflammatory regulators (NF-κß, IL-6 and CRP) were similar to control rats. Repeated infusions of LDL-DHA nanoparticles did not alter liver glutathione content or the lipid profile in the treated rats. The DHA extracted by the liver was preferentially metabolized to the anti-inflammatory DHA-derived mediator, protectin DX. CONCLUSION: Our findings indicate that repeated HAI of LDL-DHA nanoparticles is not only well tolerated and safe in the rat, but may also be protective to the liver.

Competing teammates: food in New Zealand sports settings.

Despite the global popularity of sport, we know surprisingly little about food in sports settings. This two-phased study analysed the foods available in New Zealand sports settings. Phase one included a systematic literature review and 18 interviews with key informants from national and regional sporting organizations. Phase two involved 37 key informant interviews with stakeholders from two exemplar sports, rugby and netball and direct observations at netball and rugby venues. This study found most foods and beverages at New Zealand sports events were energy-dense and nutrient-poor. Caterers' control over food provision, socio-cultural attitudes which view unhealthy foods as normal, and a dominant profit motive, appear to be the key factors influencing the food environment in sports settings. Food environments in sport settings provide frequent opportunities to purchase and consume energy-dense, nutrient-poor foods. The research shows we have competing players in the sports context-unhealthy food and healthy physical activity. Achieving sustainable healthy change in sports settings will be challenging when the prevailing attitude normalizes the unhealthy environment. Nutrition policies in sports clubs are urgently needed to increase the availability of healthy food. This requires support from health agencies and leadership from national sports organizations. Given the international nature of the food industry and sport, these findings from New Zealand may assist other countries to better understand the nature of food in sport and adopt appropriate interventions to reduce the obesogenic environment that is sport.

Priapism in a Fabry disease mouse model is associated with upregulated penile nNOS and eNOS expression.

Fabry disease is a glycosphingolipidosis caused by deficient activity of α-galactosidase A; it is one of a few diseases that are associated with priapism, an abnormal prolonged erection of the penis. The goal of this study was to investigate the pathogenesis of Fabry disease-associated priapism in a mouse model of the disease. We found that Fabry mice develop late-onset priapism. Neuronal nitric oxide synthase (nNOS), which was predominantly present as the 120-kDa N-terminus-truncated form, was significantly upregulated in the penis of 18-month-old Fabry mice compared to wild type controls (~fivefold). Endothelial NOS (eNOS) was also upregulated (~twofold). NO level in penile tissues of Fabry mice was significantly higher than wild type controls at 18 months. Gene transfer-mediated enzyme replacement therapy reversed abnormal nNOS expression in the Fabry mouse penis. The penile nNOS level was restored by antiandrogen treatment, suggesting that hyperactive androgen receptor signaling in Fabry mice may contribute to nNOS upregulation. However, the phosphodiesterase-5A expression level and the adenosine content in the penis, which are known to play roles in the development of priapism in other etiologies, were unchanged in Fabry mice. In conclusion, these data suggested that increased nNOS (and probably eNOS) content and the consequential elevated NO production and high arterial blood flow in the penis may be the underlying mechanism of priapism in Fabry mice. Furthermore, in combination with previous findings, this study suggested that regulation of NOS expression is susceptible to α-galactosidase A deficiency, and this may represent a general pathogenic mechanism of Fabry vasculopathy.

Activation of cyclic GMP-AMP synthase by self-DNA causes autoimmune diseases.

TREX1 is an exonuclease that digests DNA in the cytoplasm. Loss-of-function mutations of TREX1 are linked to Aicardi-Goutieres Syndrome (AGS) and systemic lupus erythematosus (SLE) in humans. Trex1(-/-) mice exhibit autoimmune and inflammatory phenotypes that are associated with elevated expression of interferon (IFN)-induced genes (ISGs). Cyclic GMP-AMP (cGAMP) synthase (cGAS) is a cytosolic DNA sensor that activates the IFN pathway. Upon binding to DNA, cGAS is activated to catalyze the synthesis of cGAMP, which functions as a second messenger that binds and activates the adaptor protein STING to induce IFNs and other cytokines. Here we show that genetic ablation of cGas in Trex1(-/-) mice eliminated all detectable pathological and molecular phenotypes, including ISG induction, autoantibody production, aberrant T-cell activation, and lethality. Even deletion of just one allele of cGas largely rescued the phenotypes of Trex1(-/-) mice. Similarly, deletion of cGas in mice lacking DNaseII, a lysosomal enzyme that digests DNA, rescued the lethal autoimmune phenotypes of the DNaseII(-/-) mice. Through quantitative mass spectrometry, we found that cGAMP accumulated in mouse tissues deficient in Trex1 or DNaseII and that this accumulation was dependent on cGAS. These results demonstrate that cGAS activation causes the autoimmune diseases in Trex1(-/-) and DNaseII(-/-) mice and suggest that inhibition of cGAS may lead to prevention and treatment of some human autoimmune diseases caused by self-DNA.

Engaging Without Exposing: Use of a Fictional Character to Facilitate Mental Health Talk in Focus Groups With Men Who Have Been Subject to the Criminal Justice System.

In an effort to encourage men with experience of being subject to the criminal justice system to contribute to focus group discussions on the sensitive topic of mental health, while also doing our utmost to protect them from discomfort or risk of exploitation, we used a novel technique involving the creation of a fictional character, supplemented by an audio-recorded vignette. We studied the role played by this technique in achieving our stated aims of "engaging without exposing." In this article, we report on the use of this technique in three focus groups, showing how in very different ways it shaped the interaction between participants and generated crucial insights into the lives and service needs of each group. We conclude that the technique may lend itself to being used in focus groups with other marginalized or seldom-heard populations.

Blocking hyperactive androgen receptor signaling ameliorates cardiac and renal hypertrophy in Fabry mice.

Fabry disease is caused by deficient activity of lysosomal enzyme α-galactosidase A. The enzyme deficiency results in intracellular accumulation of glycosphingolipids, leading to a variety of clinical manifestations including hypertrophic cardiomyopathy and renal insufficiency. The mechanism through which glycosphingolipid accumulation causes these manifestations remains unclear. Current treatment, especially when initiated at later stage of the disease, does not produce completely satisfactory results. Elucidation of the pathogenesis of Fabry disease is therefore crucial to developing new treatments. We found increased activity of androgen receptor (AR) signaling in Fabry disease. We subsequently also found that blockade of AR signaling either through castration or AR-antagonist prevented and reversed cardiac and kidney hypertrophic phenotype in a mouse model of Fabry disease. Our findings implicate abnormal AR pathway in the pathogenesis of Fabry disease and suggest blocking AR signaling as a novel therapeutic approach.

Patients' use and views of real-time feedback technology in general practice.

BACKGROUND: There is growing interest in real-time feedback (RTF), which involves collecting and summarizing information about patient experience at the point of care with the aim of informing service improvement. OBJECTIVE: To investigate the feasibility and acceptability of RTF in UK general practice. DESIGN: Exploratory randomized trial. SETTING/PARTICIPANTS: Ten general practices in south-west England and Cambridgeshire. All patients attending surgeries were eligible to provide RTF. INTERVENTION: Touch screens were installed in waiting areas for 12 weeks with practice staff responsible for encouraging patients to provide RTF. All practices received fortnightly feedback summaries. Four teams attended a facilitated reflection session. OUTCOMES: RTF 'response rates' among consulting patients were estimated, and the representativeness of touch screen users were assessed. The frequency of staff-patient interactions about RTF (direct observation) and patient views of RTF (exit survey) were summarized. Associated costs were collated. RESULTS: About 2.5% consulting patients provided RTF (range 0.7-8.0% across practices), representing a mean of 194 responses per practice. Patients aged above 65 were under-represented among touch screen users. Receptionists rarely encouraged RTF but, when this did occur, 60% patients participated. Patients were largely positive about RTF but identified some barriers. Costs per practice for the twelve-week period ranged from £1125 (unfacilitated team-level feedback) to £1887 (facilitated team ± practitioner-level feedback). The main cost was the provision of touch screens. CONCLUSIONS: Response rates for RTF were lower than those of other survey modes, although the numbers of patients providing feedback to each practice were comparable to those achieved in the English national GP patient survey. More patients might engage with RTF if the opportunity were consistently highlighted to them.

The Sustainable Kidney Care Foundation's contribution to the improvement of AKI management in developing countries using peritoneal dialysis.

Professional organizations, such as kidney foundations, have been active for over half a century in the field of nephrology, serving as the basic institutions for advocacy, disease education, prevention, and treatment. These organizations have focused efforts in four areas: supporting the training of clinical specialists, raising awareness about kidney disease, improving patient outcomes, and organizing continuing medical education. These activities, while essential for the success of nephrology organizations, do not usually initiate renal service programs in the neediest of places. To remedy the lack of renal programs in many developing countries, the Sustainable Kidney Care Foundation (SKCF) was founded with the objective of establishing treatment programs for acute kidney injury (AKI) in areas of the world where none exist. Today SKCF is active in 5 sub-Saharan African countries and is growing.

Saliva Urea Nitrogen Continuously Reflects Blood Urea Nitrogen after Acute Kidney Injury Diagnosis and Management: Longitudinal Observational Data from a Collaborative, International, Prospective, Multicenter Study.

BACKGROUND: Acute kidney injury (AKI) is a growing global concern and often reversible. Saliva urea nitrogen (SUN) measured by a dipstick may allow rapid diagnosis. We studied longitudinal agreement between SUN and blood urea nitrogen (BUN) and the diagnostic performance of both. METHODS: Agreement between SUN and BUN and diagnostic performance to diagnose AKI severity in AKI patients in the United States and Brazil were studied. Bland-Altman analysis and linear mixed effects models were employed to test the agreement between SUN and BUN. Receiver operating characteristics statistics were used to test the diagnostic performance to diagnose AKI severity. RESULTS: We found an underestimation of BUN by SUN, decreasing with increasing BUN levels in 37 studied patients, consistent on all observation days. The diagnostic performance of SUN (AUC 0.81, 95% CI 0.63-0.98) was comparable to BUN (AUC 0.85, 95% CI 0.71-0.98). CONCLUSION: SUN reflects BUN especially in severe AKI. It also allows monitoring treatment responses. Video Journal Club 'Cappuccino with Claudio Ronco' at http://www.karger.com/?doi=445041.

Teaching basic lung isolation skills on human anatomy simulator: attainment and retention of lung isolation skills.

BACKGROUND: Lung isolation skills, such as correct insertion of double lumen endobronchial tube and bronchial blocker, are essential in anesthesia training; however, how to teach novices these skills is underexplored. Our aims were to determine (1) if novices can be trained to a basic proficiency level of lung isolation skills, (2) whether video-didactic and simulation-based trainings are comparable in teaching lung isolation basic skills, and (3) whether novice learners' lung isolation skills decay over time without practice. METHODS: First, five board certified anesthesiologist with experience of more than 100 successful lung isolations were tested on Human Airway Anatomy Simulator (HAAS) to establish Expert proficiency skill level. Thirty senior medical students, who were naive to bronchoscopy and lung isolation techniques (Novice) were randomized to video-didactic and simulation-based trainings to learn lung isolation skills. Before and after training, Novices' performances were scored for correct placement using pass/fail scoring and a 5-point Global Rating Scale (GRS); and time of insertion was recorded. Fourteen novices were retested 2 months later to assess skill decay. RESULTS: Experts' and novices' double lumen endobronchial tube and bronchial blocker passing rates showed similar success rates after training (P >0.99). There were no differences between the video-didactic and simulation-based methods. Novices' time of insertion decayed within 2 months without practice. CONCLUSION: Novices could be trained to basic skill proficiency level of lung isolation. Video-didactic and simulation-based methods we utilized were found equally successful in training novices for lung isolation skills. Acquired skills partially decayed without practice.

Saliva urea nitrogen dipstick - a novel bedside diagnostic tool for acute kidney injury.

BACKGROUND AND AIMS: Measurement of saliva urea nitrogen (SUN) may be valuable in the screening of kidney failure. Here we evaluate the diagnostic performance of SUN dipsticks in patients with acute kidney injury (AKI). MATERIAL AND METHODS: We measured SUN and blood urea nitrogen (BUN) in hospitalized patients diagnosed with AKI based on Acute Kidney Injury Network (AKIN)-criteria. After collection, saliva was transferred to a colorimetric SUN dipstick. We then compared the resultant test-pad color to six standardized color fields indicating SUN of 5 - 14 (#1), 15 - 24 (#2), 25 - 34 (#3), 35 - 54 (#4), 55 - 74 (#5), and ≥ 75 (#6) mg/dL, respectively. We assessed the performance of SUN and BUN to discriminate AKIN 3 from earlier stages by the area under receiver operating characteristic curves (AUC ROC). RESULTS: We enrolled 44 patients (59.5 ± 18 years, 58% female; pre-renal AKI: 67%; renal 24%; post-renal 9%) in AKIN stages 1 (59%), 2 (16%), and 3 (25%). SUN and BUN levels were correlated (Spearman rank Rs = 0.69; p < 0.001, n = 44) with the highest correlation in AKIN 1 (Rs = 0.63, p = 0.001, n = 26). SUN allowed a significant discrimination of AKIN 3 from earlier stages (AUC ROC 0.91; 95% CI 0.80 - 1.0), which was comparable to the diagnostic performance of BUN (AUC ROC 0.90; 95% CI 0.78 - 1.0). CONCLUSIONS: SUN dipsticks allow the discrimination of AKIN 3 from earlier AKI stages. This low-technology approach may aid the screening of severe AKI in areas where laboratory resources are scarce.

Investigating the long-term consequences of adverse medical events among older adults.

OBJECTIVE: To investigate the long-term effect of medically serious adverse medical events (AMEs) among older adults. METHODS: We linked nationally representative survey and claims data from the Medicare Current Beneficiary Survey (1998-2004) with non-response files (1999-2005) and the Area Resource File, providing 12,541 beneficiaries with 428,373 person-months for analysis. Latent class analysis was used to assign severity status to episodes. Multinomial logistic regression was used to identify AME risk factors. The long-term consequences of AMEs on Medicare expenditures were examined by population average models. Survival analyses examined the long-term risk of death. RESULTS: Nearly 19% of participants experienced at least one AME, with 62% from outpatient claims. The risk of AMEs is greater among participants in poorer health, and increases with comorbidity and with impairment in performing activities of daily living or instrumental activities of daily living. Medicare expenditures during an AME episode increased sharply and remained higher than what would have otherwise been expected in quarters following an AME episode, and failed to return to pre-AME expenditure levels. Differences in survival rates were observable long after the AME episode concluded, with only 55% of the patients sustaining an AME surviving to the end of the study. In contrast, nearly 80% of those without an AME were estimated to have survived. CONCLUSIONS: The impacts of AMEs are observable over extended periods of time and are associated with considerable excess mortality and costs. Efforts to monitor and prevent AMEs in both acute care and outpatient settings are warranted.

Comparison of bioimpedance techniques to detect changes in fluid status in hemodialysis patients.

UNLABELLED: Bioimpedance (BI) is maturing as a clinical technique for assessing fluid volume status. The aim of this study was to compare the sensitivity of four BI methods to detect changes in fluid status in hemodialysis patients. METHODS: Forty-five patients were studied twice in the same week, i.e. once after the long and short interdialytic intervals, respectively. The four BI methods used were: (a) calf normalized resistivity (CNR) at a 5-kHz frequency, (b) whole-body multifrequency BI spectroscopy (MF-BIS) to estimate the normal hydration weight (NHWWBM), (c) whole-body MF-BIS to estimate the ratio of extracellular volume to total body water (wECV/wTBW), and (d) whole-body single-frequency (50 kHz) BI analysis to compute the ratio of ECV (sfECV) to TBW (sfTBW). RESULTS: The relationship (slope of the regressive line) between relative changes (%) in the above mentioned four BI parameters and differences in weight (kg) was most pronounced with CNR (5.2 ± 1.6%/kg), followed by wECV/wTBW (1.7 ± 0.7%/kg) and NHWWBM (0.73 ± 0.2%/kg). Changes in sfECV/sfTBW and differences in weight were not correlated. CONCLUSIONS: CNR is more sensitive than whole-body BIS for detecting differences in fluid status.

ED use by older adults attempting suicide.

OBJECTIVE: The objective of this study was to describe patterns of older adult patient visits to emergency departments (EDs) for self-harm and suicide-related injuries. METHODS: A retrospective, secondary data analysis of the Nationwide Emergency Department Sample was conducted. Nationally representative estimates of patient visits by older adults attempting suicide were calculated using available sampling weights. Population estimates were calculated using estimates from the US Census Bureau. RESULTS: Findings suggest that 22,444 ED patient visits were made by adults aged 65 years and older for suicide-related injuries, representing an estimated population rate of 63 ED patient visits per 100000 adults aged 65 years and older, with nearly half of all visits involving substance use. Total ED and hospital charges exceeded $353.9 million. CONCLUSIONS: Effort is needed to better integrate and deliver suicide screening and support services in the ED, while also connecting at-risk older adults with mental health services before and after the ED encounter.