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In 2008, Steinhart Aquarium at the California Academy of Sciences in San Francisco, CA, USA, unveiled a brand new facility with the 212,000 gallon Philippine Coral Reef habitat as its iconic centerpiece. Designing and managing a system that consists of a living reef this large, with associated invertebrates and teleosts, has been a challenge for the aquarium’s husbandry and veterinary teams. Establishing appropriate lighting, water quality, and flow has required a scientific approach and resulting adjustments to the original habitat design. The medical management of reef species has required an in-habitat approach as well as trial-and-error therapeutics. Determining the criteria for assessing the welfare of corals and other tank inhabitants has been a shifting process that has heavily utilized photography-tracked changes and other quantitative parameters as baselines. This report details the successes and limitations of establishing the environmental and veterinary management of this mesocosm. The consideration of interplay between species is discussed, and recommendations for monitoring welfare both on an individual and tank level are made. Making adjustments for the benefit of the exhibit and its animals is key for the unique aquarium displays that comprise captive coral reef systems.

Introduction: Cure Glomerulonephropathy (CureGN) is an observational cohort study of patients with minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), membranous nephropathy (MN), or IgA nephropathy. We developed a conventional, consensus-based scoring system to document pathologic features for application across multiple pathologists and herein describe the protocol, reproducibility, and correlation with clinical parameters at biopsy. Methods: Definitions were established for glomerular, tubular, interstitial, and vascular lesions evaluated semiquantitatively using digitized light microscopy slides and electron micrographs, and reported immunofluorescence. Cases with curated pathology materials as of April 2019 were scored by a randomly assigned pathologist, with at least 10% of cases scored by a second pathologist. Scoring reproducibility was assessed using Gwet’s agreement coefficient (AC)1 statistic and correlations with clinical variables were performed. Results: Of 800 scored biopsies (134 MCD, 194 FSGS, 206 MN, 266 IgA), 94 were scored twice (11.8%). Of 60 pathology features, 46 (76.7%) demonstrated excellent (AC1>0.8), and 12 (20.0%) had good (AC1 0.6–0.8) reproducibility. Mesangial hypercellularity scored as absent, focal, or diffuse had moderate reproducibility (AC1 = 0.58), but good reproducibility (AC1 = 0.71) when scored as absent or focal versus diffuse. The percent glomeruli scored as no lesions had fair reproducibility (AC1 = 0.34). Strongest correlations between pathologic features and clinical characteristics at biopsy included interstitial inflammation, interstitial fibrosis, and tubular atrophy with estimated glomerular filtration rate, foot process effacement with urine protein/creatinine ratio, and active crescents with hematuria. Conclusions: Most scored pathology features showed excellent reproducibility, demonstrating consistency for these features across multiple pathologists. Correlations between certain pathologic features and expected clinical characteristics show the value of this approach for future studies on clinicopathologic correlations and biomarker discovery.

In recent years, many researchers and software industries have given significant attention to the estimation of software effort. Software effort estimation is a challenge that often appears in the project of making software. To address this issue, various software expansion cost estimations have been familiarized. A basic traditional method for cost estimation is the Constructive Cost Model II (COCOMO II Model), which generates huge gradations of substantiality. The proposed COCOMO II model employs cost drivers, measures and the streak of programmes to predict the energy and the growth period of a web rise. However, the model is still lacking in terms of accuracy in effort and development time estimation. In this study, we do investigate the influence of components and attributes to achieve new better accuracy improvement on the COCOMO II model. The NASA93 dataset is used to test the suggested methodology. The system has outperformed earlier investigations and arrangements capable of imperfect data input, and further advancements in the reliability of the estimated technique.

Introduction Around the world, many organisations are working on ways to increase the use, sharing, and reuse of person-level data for research, evaluation, planning, and innovation while ensuring that data are secure and privacy is protected. As a contribution to broader efforts to improve data governance and management, in 2020 members of our team published 12 minimum specification essential requirements (min specs) to provide practical guidance for organisations establishing or operating data trusts and other forms of data infrastructure. Approach and Aims We convened an international team, consisting mostly of participants from Canada and the United States of America, to test and refine the original 12 min specs. Twenty-three (23) data-focused organisations and initiatives recorded the various ways they address the min specs. Sub-teams analysed the results, used the findings to make improvements to the min specs, and identified materials to support organisations/initiatives in addressing the min specs. Results Analyses and discussion led to an updated set of 15 min specs covering five categories: one min spec for Legal, five for Governance, four for Management, two for Data Users, and three for Stakeholder & Public Engagement. Multiple changes were made to make the min specs language more technically complete and precise. The updated set of 15 min specs has been integrated into a Canadian national standard that, to our knowledge, is the first to include requirements for public engagement and Indigenous Data Sovereignty. Conclusions The testing and refinement of the min specs led to significant additions and improvements. The min specs helped the 23 organisations/initiatives involved in this project communicate and compare how they achieve responsible and trustworthy data governance and management. By extension, the min specs, and the Canadian national standard based on them, are likely to be useful for other data-focused organisations and initiatives.

A GWAS of patients with anti-neutrophil cytoplasmic antibodies (ANCAs) found an association between proteinase-3 ANCA (PR3-ANCA) and a single nucleotide polymorphism (rs62132293) upstream of PRTN3, encoding PR3. The variant (G allele) was shown to be an expression quantitative trait locus in healthy controls, but the clinical impact remains unknown. Longitudinally followed patients with ANCA and healthy controls were genotyped. Gene expression was quantified by real-time quantitative PCR from leukocyte RNA. Plasma PR3 was quantified by ELISA. Among patients, variant carriers had elevated leukocyte PRTN3 expression compared with noncarriers (C/G vs. C/C and G/G vs. C/C). Healthy controls had low PRTN3 regardless of genotype. Myeloperoxidase (MPO) expression did not differ by genotype. PRTN3 expression correlated with circulating PR3, and variant carriers had higher plasma PR3 compared with noncarriers. Among variant carriers, there was an increased risk of relapse in patients with PR3-ANCA versus MPO-ANCA. The risk allele marked by rs62132293 is clinically significant as it is associated with increased autoantigen and may, in part, explain increased relapse in PR3-ANCA. Our results underscore the role of autoantigen availability in ANCA vasculitis.

Substantial evidence across disciplines indicates that children’s early years are critical for building the foundation for optimal development. In contexts of crisis, conflict, disaster and displacement, children are at particular risk. Primary caregivers often have fewer resources and less time to provide children with attention and necessary socio-emotional and cognitive stimulation. Risk factors for children are often multiplied in crises and conflict settings, and access to services becomes more challenging where family and social networks have been weakened and social service delivery interrupted. To address the needs of young children, we must strengthen humanitarian response. Practitioners often use humanitarian standards and guidance to systematise planning, implementation and evaluation of programming. These documents describe the broad principles and essential elements (i.e. specific benchmarks and actions) for high-quality response, reflecting best practices at a global level. We analysed the degree to which existing humanitarian standards and guidance documents explicitly recognise young children and their caregivers in humanitarian response and looked at gaps in the five areas that experts agree are essential for children’s holistic development: good health, adequate nutrition, security and safety, responsive caregiving and opportunities for early learning. We then made recommendations for achieving more comprehensive and consolidated guidance to address early childhood development in emergencies.

Recent scientific advances in ex situ system design and operation make it possible to complete gametogenic cycles of broadcast spawning corals. Breeding corals in aquaria is a critical advance for population management, particularly genetic rescue and assisted gene flow efforts. Genetic rescue projects for corals are already underway to bring threatened species into ex situ culture and propagation, thereby preserving standing genetic variation. However, while breeding corals is increasingly feasible, the consequences of the aquarium environment on the genetic and phenotypic composition of coral populations is not yet known. The aquarium environment may in itself be a selective pressures on corals, but it also presents relaxed selective pressures in other respects. In 2019 and 2020, gravid Acropora hyacinthus coral colonies were collected from Palauan reefs and shipped to the California Academy of Sciences (CAS) in San Francisco. In both years, gametes were batch-fertilized to produce larvae that were then settled and reared to recruits. As of April 2021, when they were sampled for sequencing, 23 corals produced at CAS in 2019 and 16 corals produced at CAS in 2020 had survived for two years and one year, respectively. We sequenced the full genomes of the 39 offspring corals and their 15 potential parents to a median 26x depth of coverage. We find clear differential parentage, with some parents producing the vast majority of offspring, while the majority of parents produced no surviving offspring. After scanning 12.9 million single nucleotide polymorphisms (SNPs), we found 887 SNPs that may be under selection in the aquarium environment, and we identified the genes and pathways these SNPs may affect. We present recommendations for preserving standing genetic variation in aquarium-bred corals based on the results of this pilot project.

Introduction: Lupus nephritis (LN) is an independent risk factor for venous thromboembolism (VTE). The risk of VTE has not been analyzed by International Society of Nephrology/Renal Pathology Society or World Health Organization LN class. Study goals were to measure VTE incidence in an LN patient cohort, to evaluate associations between VTE and LN class, and to investigate factors modifying associations between VTE and LN class. Methods: A retrospective analysis was performed using Glomerular Disease Collaborative Network data. Image-confirmed VTE was compared between patients with any LN class V lesion and patients with only LN class III or IV. Logistic regression was used to calculate odds ratios and 95% confidence intervals. Effect modification was assessed between main effect and covariates. Results: Our cohort consisted of 534 LN patients, 310 (58%) with class III/IV and 224 (42%) with class V with or without class III/IV, including 106 with class V alone. The VTE incidence was 62 of 534 (11.6%). The odds of VTE were not significantly different between patients with class III/IV and class V in adjusted analyses (odds ratio [OR] = 0.82, 95% confidence interval [CI] = 0.45−1.48). An age interaction was observed (P = 0.009), with increased odds of VTE with class III/IV diagnosed at a younger age (2.75, 0.90−8.41 estimated at age 16 years) and decreased odds with class III/IV diagnosed at an older age (0.23, 0.07−0.72 estimated at age 46 years), compared to class V. Conclusions: The VTE incidence was similar among patients with LN classes III/IV and V, suggesting that VTE risk is not limited to class V−related nephrotic syndrome and that age may modulate LN class-specific VTE risk.

Kakoki et al report a role for cyanocobalamin (B12) in preventing cardiomyopathy in type 1 diabetic mice, by reducing levels of reactive oxygen species and restoring IGF-1 levels via DNMT-SOCS1/3 signalling. The authors therefore propose B12 as a promising option to prevent diabetic cardiomyopathy through its cardioprotective effects.

Glomerular diseases (GDs) represent the third leading cause of end-stage kidney disease (ESKD) in the US Diabetes was excluded from the CureGN Study, an NIH/NIDDK-sponsored observational cohort study of four leading primary GDs: IgA nephropathy (IgAN), membranous nephropathy (MN), focal segmental glomerulosclerosis (FSGS), and minimal change disease (MCD). CureGN-Diabetes, an ancillary study to CureGN, seeks to understand how diabetes influences the diagnosis, treatment, and outcomes of GD. It is a multicenter, prospective cohort study, targeting an enrollment of 300 adults with prevalent type 1 or type 2 diabetes and MCD, FSGS, MN, or IgAN, with first kidney biopsy obtained within 5 years of enrollment in 80% (20% allowed if biopsy after 2010). CureGN and Transformative Research in DiabEtic NephropaThy (TRIDENT) provide comparator cohorts. Retrospective and prospective clinical data and patient-reported outcomes are obtained. Blood and urine specimens are collected at study visits annually. Kidney biopsy reports and digital images are obtained, and standardized pathologic evaluations performed. Light microscopy images are uploaded to the NIH pathology repository. Outcomes include relapse and remission rates, changes in proteinuria and estimated glomerular filtration rate, infections, cardiovascular events, malignancy, ESKD, and death. Multiple analytical approaches will be used leveraging the baseline and longitudinal data to compare disease presentation and progression across subgroups of interest. With 300 patients and an average of 3 years of follow-up, the study has 80% power to detect a HR of 1.4–1.8 for time to complete remission of proteinuria, a rate ratio for hospitalizations of 1.18–1.56 and difference in eGFR slope of 6.0–8.6 mL/min/year between two groups of 300 participants each. CureGN-Diabetes will enhance our understanding of diabetes as a modifying factor of the pathology and outcomes of GDs and support studies to identify disease mechanisms and improve patient outcomes in this understudied patient population.

Objective In March 2014, ICES launched Data & Analytic Services (DAS), expanding the access to ICES data and analytics beyond ICES scientists and analytic staff. In eight years, DAS has grown and evolved to increase high quality services offered to an expanding client base of external researchers. Approach At the inception of DAS, two services were offered to public sector researchers: data access and analytics. Data access enabled researchers to analyze coded record-level data through a secure virtual environment. Analytics, conducted by DAS staff in ICES analytic environment, provided researchers with risk-cleared summary level reports. In response to growing demand from an increasingly diverse range of researchers, ICES engaged in extensive consultations with internal and external stakeholders to re-evaluate and operationalize new services. Compliance with contractual obligations and Ontario law, organizational capacity to scale up, alignment with ICES’ mission, vision and values, were cornerstones in establishing new offerings. Results Analytic services became available to private sector researchers in June 2016. In March 2017, support for cohort and longitudinal follow-up studies became the newest service offering (researchers provided with a list of applicable individuals defined for the purposes of conducting publicly funded research). As more data assets become available to researchers, requests continue to increase in volume and complexity, particularly of projects seeking to import external data for linkage to ICES data. A second high performance computing virtual environment onboarded researchers September 2021 while the original analytic environment has undergone multiple upgrades, and will soon be fully refreshed. Regular solicitation of feedback has enabled DAS to increase staffing and diversify resources available which improves the client experience at all stages. Conclusions Since its inception, DAS has expanded from five to thirty personnel, grown and diversified its new and returning client base and has responded to demand for new services. DAS continues to provide high quality services which enable impactful research and is responsive to new opportunities for collaboration and service provision.

Background: Catheter ablation (CA) is performed in patients with atrial fibrillation (AF) to reduce symptoms and improve health-related quality of life (HRQL). Methods: This systematic review and meta-analysis of randomized controlled trials (RCTs) evaluated CA of any energy modality compared with antiarrhythmic drugs (AADs) using inverse-variance random-effects models. We searched for RCTs reporting HRQL and AF-related symptoms at 3, 6, 12, 24, 48, and 60 months after treatment as well as the number of repeat ablations. Results: Of 15,878 records, we included 13 RCTs of CA vs AADs for the analyses of HRQL, 7 RCTs for the analyses of AF-related symptoms, and 13 RCTs for the number of repeat ablations. For the HRQL analyses at 3 months, there were significant increases in both the Physical Component Summary score (3 months’ standardized mean difference = 0.58 [0.39-0.78]; P < 0.00001, I2 = 6%, 3 trials, n = 443) and the Mental Component Summary score (3 months’ standardized mean difference = 0.57 [0.37-0.77]; P < 0.00001, I2 = 0%, 3 trials, n = 443), favouring CA over AADs. These differences were sustained at 12 months but not >24 months after randomization. Similar results were seen for AF-related symptoms. The number of repeat ablations and success rates after procedure varied considerably across trials. Conclusions: Evidence from few trials suggests that CA improves physical and mental health and AF-related symptoms in the short term, but these benefits decrease with time. More trials, reporting both HRQL and AF-related symptoms, at consistent time points are needed to assess the effectiveness of CA for the treatment of AF. Résumé: Contexte: L’ablation par cathéter (AC) est réalisée chez les patients atteints de fibrillation auriculaire (FA) afin d’atténuer les symptômes et d’améliorer la qualité de vie liée à la santé (QVLS). Méthodologie: Cette revue systématique et méta-analyse d’essais contrôlés à répartition aléatoire (ECRA) a permis d’évaluer l’AC, toutes modalités énergétiques confondues, comparativement aux agents antiarythmiques (AA) à l’aide de modèles à effets aléatoires à variance inverse. Nous avons recherché les ECRA qui faisaient état de la QVLS et des symptômes liés à la FA à 3, 6, 12, 24, 48 et 60 mois après le traitement et qui précisaient le nombre d’ablations répétées. Résultats: À partir de 15 878 entrées, nous avons retenu 13 ECRA comparant l’AC à des AA pour l’analyse de la QVLS, 7 autres pour l’analyse des symptômes liés à la FA et 13 de plus aux fins de calcul du nombre d’ablations répétées. L’analyse de la QVLS à 3 mois indique une augmentation significative du score sommaire de la composante physique (différence moyenne normalisée à 3 mois = 0,58 [0,39-0,78]; p < 0,00001, I2 = 6 %, 3 essais, n = 443) et du score sommaire de la composante mentale (différence moyenne normalisée à 3 mois = 0,57 [0,37-0,77]; p < 0,00001, I2 = 0 %, 3 essais, n = 443), faisant pencher la balance en faveur de l’AC plutôt que des AA. Ces différences persistaient à 12 mois après la répartition aléatoire, mais pas aux moments d’évaluation ultérieurs (24, 48 et 60 mois). Des résultats similaires ont été observés au chapitre des symptômes liés à la FA. Le nombre d’ablations répétées et le taux de réussite postopératoire variaient considérablement d’un essai à l’autre. Conclusions: Les données probantes de quelques essais donnent à penser que l’AC améliore la santé physique et mentale ainsi que les symptômes liés à la FA à court terme, mais que ces bienfaits s’atténuent avec le temps. D’autres essais faisant état de la QVLS et des symptômes liés à la FA, à des moments d’évaluation constants, devront être réalisés pour évaluer l’efficacité réelle de l’AC dans le traitement de la FA.

Introduction: Venous thromboembolism (VTE) is a life-threatening complication of anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis whose mechanism remains incompletely elucidated. We tested the hypothesis that elevated microparticle tissue factor activity (MPTFa) or anti-plasminogen antibodies (anti-Plg) may identify patients at risk for VTE. Methods: In this prospective study, patients were enrolled during active disease and followed longitudinally. Twelve patients who experienced a VTE (VTEpos) were compared with patients without VTE (VTEneg, n = 29) and healthy controls (HC, n = 70). MPTFa, anti-Plg, interleukin-6, high-sensitivity C-reactive protein (hs-CRP), D-dimer, serum creatinine, and serum albumin were assessed. Fisher’s exact tests and Wilcoxon tests compared categorical and continuous variables, respectively. Cox regression for time to VTE or last follow-up was performed. Results: VTEpos patients had higher MPTFa (peak median = 14.0, interquartile range = 4.3–36.6) than HC (0, 0–3.5) and VTEneg patients (0, 0–1.4). In time-to-event analysis, MPTFa was associated with VTE when measured during both active disease (hazard ratio [HR]; 95% confidence interval [CI]: 1.04; 1.01–1.08) and remission (1.4; 1.11–1.77). Anti-Plg during remission was also associated with VTE (1.17; 1.03–1.33). Each g/dl decrease of serum albumin was associated with a 4-fold increase in VTE risk (4.4; 1.5–12.9). Adjusting for estimated glomerular filtration rate (eGFR), anti-Plg during remission remained significantly associated with VTE. Conclusion: Elevated MPTFa and increased anti-Plg in remission are strong indicators of VTE independent of renal function. Association of anti-Plg during remission with VTE implies hypercoagulability even during disease quiescence. Hypoalbuminemia strongly portends VTE risk, which is a novel finding in ANCA vasculitis. A thrombotic signature would allow improved management of patients to minimize VTE risk and complications of anticoagulation. Keywords: autoimmunity, biomarkers, plasminogen, thrombosis, tissue factor, venous thrombosis

Abstract We review the physics opportunities of the Future Circular Collider, covering its e+e-, pp, ep and heavy ion programmes. We describe the measurement capabilities of each FCC component, addressing the study of electroweak, Higgs and strong interactions, the top quark and flavour, as well as phenomena beyond the Standard Model. We highlight the synergy and complementarity of the different colliders, which will contribute to a uniquely coherent and ambitious research programme, providing an unmatchable combination of precision and sensitivity to new physics.

Introduction: In antineutrophil cytoplasmic antibody-associated (ANCA) vasculitis, relapse risk and long-term immunosuppressive therapy are problematic. Stopping immunotherapy has not been well described. Methods: The Glomerular Disease Collaborative Network ANCA vasculitis inception cohort was evaluated. Patients who stopped all immunotherapy and those continuously on immunotherapy (≥2 years) were included. Time to first period off therapy was modeled with end-stage kidney disease and death as competing risks to understand influences of stopping therapy. Cause-specific hazard ratios (HRs) with 95% confidence intervals (CI) and P values are reported. Models controlled for age, sex, ANCA specificity, organ involvement, diagnosis era, and treatments (yes/no). Repeated events analysis was used to assess the time-dependent variable of time off treatment on recurrent relapse with HRs, 95% CIs, and P values are reported (same control variables without treatments). Results: In 427 patients, 277 (65%) stopped therapy (median 20 months from initial induction); 14% for ≥2 different periods of time and 23% for periods ≥5 years. In multivariable models of time to discontinuation of treatment, women (HR 1.33; 95% CI 1.04–1.70; P = 0.024) and those treated with pulse methylprednisolone (HR 1.39; 95% CI 1.05–1.84; P = 0.020) were more likely to stop. The time-dependent variable of time off treatment was associated with fewer recurrent relapses (HR 0.51; 95% CI 0.41–0.63; P < 0.001). Conclusions: Stopping immunotherapy was common. Women and those treated with methylprednisolone stop treatment more often, but underlying mechanisms are unknown. Stopping treatment was associated with fewer relapses, suggesting that even without guidelines there may be benefits without an untoward detriment of relapse. Keywords: ANCA, relapse, stopping therapy, time off therapy, vasculitis

Background: Multiple Imputation (MI) is known as an effective method for handling missing data in public health research. However, it is not clear that the method will be effective when the data contain a high percentage of missing observations on a variable. Methods: Using data from “Predictive Study of Coronary Heart Disease” study, this study examined the effectiveness of multiple imputation in data with 20% missing to 80% missing observations using absolute bias (|bias|) and Root Mean Square Error (RMSE) of MI measured under Missing Completely at Random (MCAR), Missing at Random (MAR), and Not Missing at Random (NMAR) assumptions. Results: The |bias| and RMSE of MI was much smaller than of the results of CCA under all missing mechanisms, especially with a high percentage of missing. In addition, the |bias| and RMSE of MI were consistent regardless of increasing imputation numbers from M=10 to M=50. Moreover, when comparing imputation mechanisms, MCMC method had universally smaller |bias| and RMSE than those of Regression method and Predictive Mean Matching method under all missing mechanisms. Conclusion: As missing percentages become higher, using MI is recommended, because MI produced less biased estimates under all missing mechanisms. However, when large proportions of data are missing, other things need to be considered such as the number of imputations, imputation mechanisms, and missing data mechanisms for proper imputation.

Introduction There is widespread enthusiasm to improve health through the application of artificial intelligence and machine learning (AI/ML) methods to large population-level health datasets. Achieving this may require successful collaboration between institutions as well as between computer scientists (CS), machine learning researchers (MLR) and health service researchers (HSR). The objective is to describe lessons learned in creating the Health Artificial Data and Analysis Platform (HAIDAP) in Ontario, Canada. Objectives and Approach A partnership between a HSR institute (ICES), an AI/ML institute (Vector) and a high-performance computing center (HPC4H) was initiated in 2017 to enable the application of AI/ML methods to population-level health data for the province of Ontario (population 14M). The HAIDAP was launched in 2019. We describe lessons learned (and being learned) following the HAIDAP’s launch. Results Major learnings include: 1)importance of institutional partnerships and alignment with institutional strategies; 2)potential of joint institutional risk-sharing models; 3)need for scientific collaborations bridging disciplines around joint research projects; 4)sensitivity to different scientific cultures (e.g., academic prestige of conference proceedings for MLR vs journal publications for HSR; traditional statistical vs. ML model assumptions); 5)differences in research timeline expectations; 6)different experience with and expectations for access to de-identified routinely collected data (e.g., need for research ethics committee project approvals and privacy impact assessments); 7)developing data access models that enable greater flexibility (e.g. importing code or using open source tools); 8) broadening data access models to allow modern high-dimensional exploratory data analysis; 9)obtaining support of information/privacy regulator; 10)hardware is (relatively) easy part compared to other success factors. Conclusion / Implications The HAIDAP has enabled multi-disciplinary collaborations and novel AI/ML research of Ontario’s population-level health data. Collectively we have learned that additional effort is required to develop systems and processes enabling more efficient access to data and analytic tools for the analysis of administrative health data.

The development of vaccines against flaviviruses, including Zika virus (ZIKV) and dengue virus (DENV), continues to be a major challenge, hindered by the lack of efficient and reliable methods for screening neutralizing activity of sera or antibodies. To address this need, we previously developed a plasmid-based, replication-incompetent DENV reporter virus particle (RVP) production system as an efficient and safe alternative to the Plaque Reduction Neutralization Test (PRNT). As part of the response to the 2015–2016 ZIKV outbreak, we developed pseudo-infectious ZIKV RVPs by modifying our DENV RVP system. The use of ZIKV RVPs as critical reagents in human clinical trials requires their further validation using stability and reproducibility metrics for large-scale applications. In the current study, we validated ZIKV RVPs using infectivity, neutralization, and enhancement assays with monoclonal antibodies (MAbs) and human ZIKV-positive patient serum. ZIKV RVPs are antigenically equivalent to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of live virus replication assays. We demonstrate reproducible neutralization titer data (NT50 values) across different RVP production lots, volumes, time frames, and laboratories. We also show RVP stability across experimentally relevant time intervals and temperatures. Our results demonstrate that ZIKV RVPs provide a safe, high-throughput, and reproducible reagent for large-scale, long-term studies of neutralizing antibodies and sera, which can facilitate large-scale screening and epidemiological studies to help expedite ZIKV vaccine development. Author summary ZIKV is a mosquito-borne virus that can cause severe birth defects and other disorders. Large outbreaks of ZIKV occurred in 2015 and 2016 and there are still no drugs or vaccines available to protect against ZIKV infection. Vaccine development has been hindered by the lack of safe and efficient methods to screen ZIKV neutralizing properties of patient sera or antibodies, especially in the context of large clinical trials. To address this unmet need, we developed and validated the use of ZIKV reporter virus particles (RVPs), a safe, high-throughput, and quantitative alternative to using live virus for neutralization studies. We show that ZIKV RVPs are stable, show lot-to-lot consistency, and provide reproducible neutralization data that is suitable for the large-scale studies needed for development of a ZIKV vaccine, epidemiologic surveillance, and high-throughput screening.

Introduction Pain negatively affects the health-related quality of life (HRQL) of adolescents with cancer. The Pain Squad+ smartphone-based application (app), has been developed to provide adolescents with real-time pain self-management support. The app uses a validated pain assessment and personalised pain treatment advice with centralised decision support via a registered nurse to enable real-time pain treatment in all settings. The algorithm informing pain treatment advice is evidence-based and expert-vetted. This trial will longitudinally evaluate the impact of Pain Squad+, with or without the addition of nurse support, on adolescent health and cost outcomes.Methods and analysis This will be a pragmatic, multicentre, waitlist controlled, 3-arm parallel-group superiority randomised trial with 1:1:1 allocation enrolling 74 adolescents with cancer per arm from nine cancer centres. Participants will be 12 to 18 years, English-speaking and with ≥3/10 pain. Exclusion criteria are significant comorbidities, end-of-life status or enrolment in a concurrent pain study. The primary aim is to determine the effect of Pain Squad+, with and without nurse support, on pain intensity in adolescents with cancer, when compared with a waitlist control group. The secondary aims are to determine the immediate and sustained effect over time of using Pain Squad+, with and without nurse support, as per prospective outcome measurements of pain interference, HRQL, pain self-efficacy and cost. Linear mixed models with baseline scores as a covariate will be used. Qualitative interviews with adolescents from all trial arms will be conducted and analysed.Ethics and dissemination This trial is approved by the Hospital for Sick Children Research Ethics Board. Results will provide data to guide adolescents with cancer and healthcare teams in treating pain. Dissemination will occur through partnerships with stakeholder groups, scientific meetings, publications, mass media releases and consumer detailing.Trial registration number NCT03632343 (ClinicalTrials.gov).

Abstract Background Fabry disease is frequently characterized by gastrointestinal symptoms, including diarrhea. Migalastat is an orally-administered small molecule approved to treat the symptoms of Fabry disease in patients with amenable mutations. Methods We evaluated minimal clinically important differences (MCID) in diarrhea based on the corresponding domain of the patient-reported Gastrointestinal Symptom Rating Scale (GSRS) in patients with Fabry disease and amenable mutations (N = 50) treated with migalastat 150 mg every other day or placebo during the phase 3 FACETS trial (NCT00925301). Results After 6 months, significantly more patients receiving migalastat versus placebo experienced improvement in diarrhea based on a MCID of 0.33 (43% vs 11%; p = .02), including the subset with baseline diarrhea (71% vs 20%; p = .02). A decline in kidney peritubular capillary globotriaosylceramide inclusions correlated with diarrhea improvement; patients with a reduction > 0.1 were 5.6 times more likely to have an improvement in diarrhea than those without (p = .031). Conclusions Migalastat was associated with a clinically meaningful improvement in diarrhea in patients with Fabry disease and amenable mutations. Reductions in kidney globotriaosylceramide may be a useful surrogate endpoint to predict clinical benefit with migalastat in patients with Fabry disease. Trial registration NCT00925301; June 19, 2009.

Abstract Background Altered expression of mRNA splicing factors occurs with ageing in vivo and is thought to be an ageing mechanism. The accumulation of senescent cells also occurs in vivo with advancing age and causes much degenerative age-related pathology. However, the relationship between these two processes is opaque. Accordingly we developed a novel panel of small molecules based on resveratrol, previously suggested to alter mRNA splicing, to determine whether altered splicing factor expression had potential to influence features of replicative senescence. Results Treatment with resveralogues was associated with altered splicing factor expression and rescue of multiple features of senescence. This rescue was independent of cell cycle traverse and also independent of SIRT1, SASP modulation or senolysis. Under growth permissive conditions, cells demonstrating restored splicing factor expression also demonstrated increased telomere length, re-entered cell cycle and resumed proliferation. These phenomena were also influenced by ERK antagonists and agonists. Conclusions This is the first demonstration that moderation of splicing factor levels is associated with reversal of cellular senescence in human primary fibroblasts. Small molecule modulators of such targets may therefore represent promising novel anti-degenerative therapies.

Abstract We discuss the determination of the CKM angle $$\alpha $$ α using the non-leptonic two-body decays $$B\rightarrow \pi \pi $$ B → π π , $$B\rightarrow \rho \rho $$ B → ρ ρ and $$B\rightarrow \rho \pi $$ B → ρ π using the latest data available. We illustrate the methods used in each case and extract the corresponding value of $$\alpha $$ α . Combining all these elements, we obtain the determination $$\alpha _\mathrm{dir}={({86.2}_{-4.0}^{+4.4} \cup {178.4}_{-5.1}^{+3.9})}^{\circ }$$ α dir = ( 86.2 - 4.0 + 4.4 ∪ 178.4 - 5.1 + 3.9 ) ∘ . We assess the uncertainties associated to the breakdown of the isospin hypothesis and the choice of the statistical framework in detail. We also determine the hadronic amplitudes (tree and penguin) describing the QCD dynamics involved in these decays, briefly comparing our results with theoretical expectations. For each observable of interest in the $$B\rightarrow \pi \pi $$ B → π π , $$B\rightarrow \rho \rho $$ B → ρ ρ and $$B\rightarrow \rho \pi $$ B → ρ π systems, we perform an indirect determination based on the constraints from all the other observables available and we discuss the compatibility between indirect and direct determinations. Finally, we review the impact of future improved measurements on the determination of $$\alpha $$ α .

Overall objectives or goal Many health data linkage ecosystems across the world have designed and implemented secure data analysis environments as one of their controls to protect patient privacy and confidentiality. These have been shaped by local legislation and data governance policies, available IT infrastructure and resources, and the skills and imagination of their architects. However, at present their various features and functionalities have not been reviewed, synthesised or contrasted. Burton et al [1] have proposed 12 criteria for Data Safe Havens in health and healthcare, which they conceptualise broadly as encompassing data governance and ethics, quality and curation of data repositories, and data security. Under this definition, secure analysis environments, which may or may not be integrated with data repositories, are a component of a Data Safe Haven, addressing the criterion “Appropriate secure access to individually identifying data”. To guide those building and operating these environments, and data custodians and stewards who need to assess their fitness-for-purpose, it would be of great value to discuss and agree an aggregate term (e.g. “Secure Data Lab”) that describes them, and to develop a more detailed set of criteria for what entails “Appropriate secure access” to linked health data. The goal of this session is to describe and document the approaches that have been taken by flagship secure data analysis environments internationally, including their approaches to authentication, assigning permissions, managing the ingress and egress of files and auditing transactions, and their responses to emerging opportunities, including cloud computing and national and international data sharing. We will explore how the interplay of physical, technical and procedural controls have been combined to create existing models, and the extent to which these can balance each other and be applied with flexibility depending on perceived risk and regimes. Session structure Prior to the session, we will develop a draft set of criteria for “Appropriate secure access” to linked health data. The session will comprise presentations describing existing secure analysis environments against the draft criteria, followed by a facilitated discussion. The secure data analysis environments that will be presented include: • UNSW Sydney E-Research Institutional Cloud Architecture (ERICA) • PopData BC Secure Research Environment (SRE) • Institute for Clinical Evaluative Sciences (ICES) Data and Analytic Virtual Environment (IDAVE) • Secure Anonymised Information Linkage (SAIL) Gateway Intended output or outcome We will write up the outcomes of the session as a scientific paper that proposes an aggregate term for secure data analysis environments for linked health data and a set of criteria for what entails “Appropriate secure access” to linked health data. Presenters and Facilitators Professor Louisa Jorm, Centre for Big Data Research in Health, UNSW Sydney, Australia Dr Tim Churches, South Western Sydney Clinical School, UNSW Sydney, Australia Professor Kim McGrail, Population Data BC, The University of British Columbia, Vancouver, Canada J. Charles Victor, Institute for Clinical Evaluative Sciences, Toronto, Canada Dr Kerina Jones, Swansea University Medical School, Wales, United Kingdom Professor David Ford, Swansea University Medical School, Wales, United Kingdom 1. Burton PR, Murtagh MJ, Boyd A, et al. Data Safe Havens in health research and healthcare. Bioinformatics 2015; 31(20): 3241–3248

Introduction The Ontario Brain Institute has developed Brain-CODE, an informatics platform, to support the acquisition, storage, management and analysis of multi-modal data. The standardized research data within Brain-CODE spans several brain disorders, allowing for integrative analyses, while also providing the opportunity to leverage existing clinical administrative data holdings through external linkages. Objectives and Approach Within Ontario, the majority of individuals who access the healthcare system have a unique identifier, the Ontario Health Insurance Plan (OHIP) number. The OHIP number can facilitate linkages with administrative data holdings, such as those at the Institute for Clinical Evaluative Sciences (ICES). Given that OBI is not permitted under Ontario’s privacy legislation to hold OHIP numbers, identifiers for consented participants are encrypted using a public key mechanism upon entry into Brain-CODE, where the private key is inaccessible. To facilitate linkages involving OHIP numbers between Brain-CODE and ICES, Brain-CODE Link software was co-developed by members of the Indoc Consortium. Results Brain-CODE Link allows a deterministic linkage between encrypted identifiers (OHIP numbers), without revealing participant identity. The same homomorphic encryption algorithm applied to identifiers upon entry to Brain-CODE, is applied to relevant identifiers within ICES data holdings. Encrypted identifiers from Brain-CODE are securely transferred to ICES, where a comparison computation calculates differences between the encrypted sets. These differences are sent to a semi-trusted third party, who has no access to the original data, to decrypt the differences using the private key. A zero difference indicates a set of matching identifiers. One of the main challenges during testing and development of Brain-CODE Link was ensuring the software was capable of scaling to a population level, performing a large number of comparisons, in a computationally efficient manner. Conclusion/Implications Ongoing pilot projects within the areas of epilepsy, neurodevelopment disorders, and neurodegeneration will be the first examples of linkages between Brain-CODE and ICES. Brain-CODE Link has successfully performed several billion test comparisons, indicating its suitability to function as a scalable privacy preserving record linkage to support comprehensive analyses.

Introduction Researchers and decision makers from across Canada use linked provincial administrative data for analysis and to address research and policy questions. Currently there are several impediments to working harmoniously across provincial boundaries. A group of academic and policy researchers are working to address these multi-jurisdictional obstacles. Objectives and Approach Researchers and data organizations from across Canada are working together as the Pan-Canadian Real-World Health Data Network PRHDN). PRHDN aims to: (1) create harmonized data, algorithms and analytic protocols, and (2) link administrative databases to other types of data, including electronic medical records, clinical trials records, “omics data” and records from pan-Canadian cohort studies. PRHDN’s vision is to construct a unified, documented infrastructure to advance pan-Canadian population-based research and analysis. This presentation incorporates material that is part of PRHDN’s response to a funding call to create national, collaborative infrastructure. Results Scientists and staff at PRHDN organizations will create three main categories of infrastructure: 1) Algorithms: Reusable processes, ideally in the form of documented code, which implement a common approach or definition, e.g. to define cases or to create derived variables; 2) Harmonized Common Data: Based on the Sentinel model, we will establish a standardized subset of harmonized common data that are analysis-ready; 3) Common Analytic Protocols: Complementing work of the Canadian Network for Observational Drug Effect Studies (CNODES), we will establish processes for distributed analysis with common analytic protocols and meta-analysis of results to provide pan-Canadian estimates. Source data would remain within jurisdictional boundaries and only aggregate results would be pooled across jurisdictions. Details of these approaches will be presented. Conclusion/Implications This initiative will improve coordinated access to distributed data from across Canada that is built once then used by many stakeholders for a variety of purposes including: research, benchmarking, performance monitoring to identify gaps and opportunities for improvement, multi-jurisdictional evaluations of novel interventions and inter-jurisdictional comparisons.

Introduction Improving the care and management of patients with diabetes, particularly those with extreme blood glucose and/or cholesterol levels, has been identified as a key priority area for healthcare in Ontario. A multi-organizational collaboration produces audit-and-feedback reports distributed to consenting primary care physicians across the province for quality improvement purposes. Objectives and Approach We examined the feasibility of linking the Ontario Laboratory Information System (OLIS), a large and nearly population-wide database of laboratory test results in Ontario, with the existing provincial audit-and-feedback reporting structure to integrate aggregated, physician-level measures of glycemic and cholesterol control among patients with diabetes. All Ontario residents alive on March 31, 2014, attached to a primary care physician, and diagnosed with diabetes for at least two years were included. These patients were linked to OLIS to extract laboratory test orders and results for glycated hemoglobin (HbA1C) and low-density lipoproteins (LDL) between April 1, 2013 and March 31, 2014. Results There were 1,108,530 diabetes patients included who were assigned to 10,085 primary care physicians. During fiscal year (FY) 2013, 70%, 64%, and 59% of diabetes patients were tested for HbA1C, LDL, and both measures, respectively. Among the 648,238 diabetes patients with at least one of each test in FY2013, 13% had a HbA1C test exceeding a threshold of 9%, 4% had a LDL test exceeding a threshold of 4 mmol/L, and 0.8% exceeded both thresholds. At the physician-level, the median (Interquartile Range) proportions of diabetes patients exceeding the testing thresholds were 12% (9%-16%) for HbA1c and 4% (2%-6%) for LDL. In a multilevel logistic regression model, there was significant between-physician variability in the proportions of diabetes patients exceeding the HbA1C (p Conclusion/Implications We developed a mechanism for integrating population-wide, clinical laboratory test results into physician audit-and-feedback reports to improve diabetes care in Ontario. Significant variation observed in the aggregated, physician-level proportions of diabetes patients testing above clinical thresholds for HbA1C and LDL highlights the importance of reporting such information to physicians.

Introduction Canadian provincial health systems have a data advantage – longitudinal population-wide data for publicly funded health services, in many cases going back 20 years or more. With the addition of high performance computing (HPC), these data can serve as the foundation for leading-edge research using machine learning and artificial intelligence. Objectives and Approach The Institute for Clinical Evaluative Sciences (ICES) and HPC4Health are creating the Ontario Data Safe Haven (ODSH) – a secure HPC cloud located within the HPC4Health physical environment at the Hospital for Sick Children in Toronto. The ODSH will allow research teams to post, access and analyze individual datasets over which they have authority, and enable linkage to Ontario administrative and other data. To start, the ODSH is focused on creating a private cloud meeting ICES’ legislated privacy and security requirements to support HPC-intensive analyses of ICES data. The first ODSH projects are partnerships between ICES scientists and machine learning. Results As of March 2018, the technological build of the ODSH was tested and completed and the privacy and security policy framework and documentation were completed. We will present the structure of the ODSH, including the architectural choices made when designing the environment, and planned functionality in the future. We will describe the experience to-date for the very first analysis done using the ODSH: the automatic mining of clinical terminology in primary care electronic medical records using deep neural networks. We will also present the plans for a high-cost user Risk Dashboard program of research, co-designed by ICES scientists and health faculty from the Vector Institute for artificial intelligence, that will make use of the ODSH beginning May 2018. Conclusion/Implications Through a partnership of ICES, HPC4Health and the Vector Institute, a secure private cloud ODSH has been created as is starting to be used in leading edge machine learning research studies that make use of Ontario’s population-wide data assets.

Overall objectives or goal Most of the organizations that use population administrative data for research purposes have internal repository of validated definitions and algorithms of their own. Many of these concepts and definitions are applicable or at least adaptable to other organizations and jurisdictions. A comprehensive National (and potentially International) Concept Dictionary could help investigators to carry out methodologically sound work using consistent and validated algorithms using a shared pool of knowledge and resources. The Institute for Clinical Evaluative Sciences (ICES) in Ontario, Canada has recently modernized its internal Concept Dictionary by adopting standard templates based on the Manitoba Centre for Health Policy (MCHP) Concept Dictionary, reviewing and updating existing content and tagging the concept entries with appropriate MeSH terms and data sources, and adding standard computer code (e.g., SAS coding) where appropriate. A SharePoint® web-based application has been developed to provide advanced tagging, searching and browsing features. We envision a wiki-based Concept Dictionary hosted on a cloud-based environment with very granular access controls to provide enough flexibility for each participating organization to control their own content. This means each organization will be able to decide on how to share their own concepts (or part of them) with the public or internal users. All content will be tagged with MeSH terms and as well with the organization’s name that initially posts each entry. Other organizations which find the same concept applicable to their own use can tag the same entry with their organization name or refer to a secondary adapted entry if adaptation to fit their data and methodologies is required. The Search feature will allow refining the search criteria by MeSH terms, data sources, and also organization/jurisdiction name. Multiple layers of access controls will allow each organization to have their own groups of users with different standard privileges such as Local Administrators, Authors and Approvers (or Publishers). The Approver (Publisher) users within each organization can publish each entry for internal or public view. This way, for example, a definition/algorithm can be viewable only within the organization until the validation process is complete, and then the entry can be made publically available, while some sections, such as computer code, can remain restricted to the organization. We will discuss challenges in developing and maintaining such a platform including the costs, governance, intellectual property rights, copyrights and liabilities for the participating organizations. The intended output or outcome We aim to use this opportunity to form a working group from the interested organizations that are ready to participate and commit in developing this collaborative platform. After the conference, there will be follow up sessions with the members of the working group to plan and develop the online application.

Introduction Integrating health and social services data is critical to understanding social determinants of health and responding to public expectations for evidence-based policies amidst changing demographics and fiscal constraint. While academia has long understood the importance of social determinants of health, real and perceived obstacles have slowed their evaluation in Ontario. Objectives and Approach This report describes how the Institute for Clinical Evaluative Sciences (ICES) and the Ministry and Community and Social Services (MCSS) have partnered to bring social services data and health data together to better understand the Ontario population and better support decision makers across various sectors. We present how ICES and MCSS tackled barriers to data access and cultural challenges to data sharing in the Ontario context, provide an overview of their unique data and research partnership - including the new collaboration research and data access platforms created, highlight research findings to date, and identify key topics of interest moving forward. Results Over the last decade, ICES and MCSS have led the way in Ontario linking health administrative and social services data. An initial single year linkage enabled the success of the Health Care Access Research and Developmental Disabilities project. This cross-sectoral initiative provided a clearer sense of how people with developmental disabilities experienced health care in Ontario. Building on this work, ICES and MCSS recently expanded their partnership bringing together 15 years of social services and health data through a broader data sharing agreement. This agreement allows greater data access to researchers. In addition, ICES and MCSS have been successful in creating a new integrated research platform that will increase the depth and quality of health and social services research and policy evaluation in Ontario. Conclusion/Implications A broader collaborative research community will now be able to answer questions of interest, do self-directed integrated data analytics and leverage respective program data expertise to tackle joint research projects. Importantly, MCSS analytics teams will now also have access to linked data on this platform to conduct their own research.

ABSTRACT All enveloped viruses, including herpesviruses, must fuse their envelope with the host membrane to deliver their genomes into target cells, making this essential step subject to interference by antibodies and drugs. Viral fusion is mediated by a viral surface protein that transits from an initial prefusion conformation to a final postfusion conformation. Strikingly, the prefusion conformation of the herpesvirus fusion protein, gB, is poorly understood. Herpes simplex virus (HSV), a model system for herpesviruses, causes diseases ranging from mild skin lesions to serious encephalitis and neonatal infections. Using cryo-electron tomography and subtomogram averaging, we have characterized the structure of the prefusion conformation and fusion intermediates of HSV-1 gB. To this end, we have set up a system that generates microvesicles displaying full-length gB on their envelope. We confirmed proper folding of gB by nondenaturing electrophoresis-Western blotting with a panel of monoclonal antibodies (MAbs) covering all gB domains. To elucidate the arrangement of gB domains, we labeled them by using (i) mutagenesis to insert fluorescent proteins at specific positions, (ii) coexpression of gB with Fabs for a neutralizing MAb with known binding sites, and (iii) incubation of gB with an antibody directed against the fusion loops. Our results show that gB starts in a compact prefusion conformation with the fusion loops pointing toward the viral membrane and suggest, for the first time, a model for gB’s conformational rearrangements during fusion. These experiments further illustrate how neutralizing antibodies can interfere with the essential gB structural transitions that mediate viral entry and therefore infectivity. IMPORTANCE The herpesvirus family includes herpes simplex virus (HSV) and other human viruses that cause lifelong infections and a variety of diseases, like skin lesions, encephalitis, and cancers. As enveloped viruses, herpesviruses must fuse their envelope with the host membrane to start an infection. This process is mediated by a viral surface protein that transitions from an initial conformation (prefusion) to a final, more stable, conformation (postfusion). However, the prefusion conformation of the herpesvirus fusion protein (gB) is poorly understood. To elucidate the structure of the prefusion conformation of HSV type 1 gB, we have employed cryo-electron microscopy to study gB molecules expressed on the surface of vesicles. Using different approaches to label gB’s domains allowed us to model the structures of the prefusion and intermediate conformations of gB. Overall, our findings enhance our understanding of HSV fusion and lay the groundwork for the development of new ways to prevent and block HSV infection.

ABSTRACT Background The Institute for Clinical Evaluative Sciences (ICES) is a not-for-profit organization that conducts research to evaluate health care delivery and outcomes. Established in 1992, ICES houses a vast and secure array of linkable, coded health-related data on more than 13 million Ontarians, including health services data, health care provider data, registries and population-based health surveys. ICES has a reputation for generating strong evidence-based knowledge to inform policy and practice, however the use of the data was restricted to ICES’ purposes. In March 2014, ICES launched the Data & Analytic Services (DAS) platform with the primary objective of increasing access to its data to publicly-funded researchers, health care providers and administrators, policymakers and students. Method DAS provides access to highly de-identified, risk-reduced datasets created from ICES’ data holdings; analytic support; and complete data analysis and report writing services. DAS also enables the importation of external data for linkage to ICES’ data holdings. Research objectives and methodology are led by the requestor and ICES analysts rely on their subject matter expertise to direct the deliverables. Results Since launch, over 200 requests from Canada, United States and United Kingdom have been adjudicated, of which 187 have been deemed feasible and eligible. Over the same period of time, ICES as an organization had over 700 active projects of which 348 were initiated, an increase in capacity of 26%. Though Toronto-based researchers represent the majority of the requests (62%), there have been requests from outside Ontario interested in comparing aspects of Ontario’s healthcare to their home province. Research topics have varied and include assessments of health care provision by sector, disease prevalence and treatment, and statistical methods. An unexpected outcome of increasing access has been the large interest from small physician groups and knowledge users who are not typically involved in research for academic purposes. Access to ICES’ data holdings provides an opportunity to examine a larger cohort of patients who share the same characteristics as their clinic patients or group. Furthermore, by enabling remote access to the data, DAS is able to leverage the capabilities of ICES’ data holdings and increase research capacity in a short period of time. Conclusion In making one of the most comprehensively linked health administrative data repositories in the world widely available to the broader research and healthcare community, DAS engages investigators involved in front-line care, stimulates new avenues of research and fosters collaboration that was previously unachievable.

Photothermal therapy using nanoparticles is a promising new approach for the treatment of cancer. The principle is to utilise plasmonic nanoparticle light interaction for efficient heat conversion. However, there are many hurdles to overcome before it can be accepted in clinical practice. One issue is a current poor characterization of the thermal dose that is distributed over the tumour region and the surrounding normal tissue. Here, we use Monte Carlo simulations of photon radiative transfer through tissue and subsequent heat diffusion calculations, to model the spatial thermal dose in a skin cancer model. We validate our heat rise simulations against experimental data from the literature and estimate the concentration of nanorods in the tumor that are associated with the heat rise. We use the cumulative equivalent minutes at 43 °C (CEM43) metric to analyse the percentage cell kill across the tumour and the surrounding normal tissue. Overall, we show that computer simulations of photothermal therapy are an invaluable tool to fully characterize thermal dose within tumour and normal tissue.

ABSTRACT Herpesvirus entry requires the viral glycoprotein triad of gB and gH/gL to carry out fusion between the virion envelope and a cellular membrane in order to release the nucleocapsid into the target cell. Herpes simplex virus (HSV) also requires glycoprotein gD to initiate the fusion cascade by binding a cell receptor such as nectin 1 or herpesvirus entry mediator (HVEM). While the structure of gB is that of a class III fusion protein, gH/gL has no features that resemble other viral fusion proteins. Instead, it is suggested that gH/gL acts as a regulator of gB. The crystal structure of HSV-2 gH/gL was obtained with a functional protein that had a deletion of 28 residues at the gH N terminus (gHΔ48/gL). Unexplainably, monoclonal antibodies (MAbs) with virus-neutralizing activity map to these residues. To reconcile these two disparate observations, we studied the ability of gHΔ48/gL to regulate fusion. Here, we show that the protein induces low (constitutive) levels of fusion by gB in the absence of gD and/or receptor. However, when gD and receptor are present, this mutant functions as well as does wild-type (wt) gH/gL for fusion. We propose that gHΔ48/gL has an intermediate structure on the pathway leading to full regulatory activation. We suggest that a key step in the pathway of fusion is the conversion of gH/gL to an activated state by receptor-bound gD; this activated gH/gL resembles gHΔ48/gL. IMPORTANCE Herpes simplex viruses (HSVs) cause many human diseases, from mild cold sores to lethal neonatal herpes. As an enveloped virus, HSV must fuse its membrane with a host membrane in order for replication to take place. The virus uses four glycoproteins for this process, gD, gB, and gH/gL, and either of two cell receptors, herpesvirus entry mediator (HVEM) and nectin 1. Although the virus can enter the cell by direct fusion at the plasma membrane or via endocytosis, the same four glycoproteins are involved. The absence of any of these proteins abolishes the entry process. Here, we show that a mutant form of gH/gL, gHΔ48/gL, can induce fusion of gB-expressing cells in the absence of gD and a gD receptor. Our study supports the concept that gB is the HSV fusogen and its activity is regulated by gH/gL.

Whisker movement has been shown to be under active control in certain specialist animals such as rats and mice. Though this whisker movement is well characterised, the role and effect of this movement on subsequent sensing is poorly understood. One method for investigating this phenomena is to generate artificial whisker deflections with robotic hardware under different movement conditions. A limitation of this approach is that assumptions must be made in the design of any artificial whisker actuators, which will impose certain restrictions on the whisker-object interaction. In this paper we present three robotic whisker platforms, each with different mechanical whisker properties and actuation mechanisms. A feature-based classifier is used to simultaneously discriminate radial distance to contact and contact speed for the first time. We show that whisker-object contact speed predictably affects deflection magnitudes, invariant of whisker material or whisker movement trajectory. We propose that rodent whisker control allows the animal to improve sensing accuracy by regulating contact speed induced touch-to-touch variability.

ABSTRACT Glycoprotein B (gB), gD, and gH/gL constitute the fusion machinery of herpes simplex virus (HSV). Prior studies indicated that fusion occurs in a stepwise fashion whereby the gD/receptor complex activates the entire process, while gH/gL regulates the fusion reaction carried out by gB. Trimeric gB is a class III fusion protein. Its ectodomain of 773 amino acids contains a membrane-proximal region (MPR) (residues 731 to 773) and two fusion loops (FLs) per protomer. We hypothesized that the highly hydrophobic MPR interacts with the FLs, thereby masking them on virions until fusion begins. To test this hypothesis, we made a series of deletion, truncation, and point mutants of the gB MPR. Although the full-length deletion mutants were expressed in transfected cells, they were not transported to the cell surface, suggesting that removal of even small stretches of the MPR was highly detrimental to gB folding. To circumvent this limitation, we used a baculovirus expression system to generate four soluble proteins, each lacking the transmembrane region and cytoplasmic tail. All retained the FLs and decreasing portions of the MPR [gB(773t) (gB truncated at amino acid 773), gB(759t), gB(749t), and gB(739t)]. Despite the presence of the FLs, all were compromised in their ability to bind liposomes compared to the control, gB(730t), which lacks the MPR. We conclude that residues 731 to 739 are sufficient to mask the FLs, thereby preventing liposome association. Importantly, mutation of two aromatic residues (F732 and F738) to alanine restored the ability of gB(739t) to bind liposomes. Our data suggest that the MPR is important for modulating the association of gB FLs with target membranes. IMPORTANCE To successfully cause disease, a virus must infect host cells. Viral infection is a highly regulated, multistep process. For herpesviruses, genetic material transfers from the virus to the target cell through fusion of the viral and host cell lipid membranes. Here, we provide evidence that the ability of the herpes simplex virus (HSV) glycoprotein B (gB) fusion protein to interact with the host membrane is regulated by its membrane-proximal region (MPR), which serves to cover or shield its lipid-associating moieties (fusion loops). This in turn prevents the premature binding of gB with host cells and provides a level of regulation to the fusion process. These findings provide important insight into the complex regulatory steps required for successful herpesvirus infection.