Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes

The hepatitis C virus (HCV) nonstructural 5A (NS5A) protein is highly phosphorylated and involved in both virus genome replication and virion assembly. We and others have identified serine 225 in NS5A to be a phosphorylation site, but the function of this posttranslational modification in the virus life cycle remains obscure. Here we describe the phenotype of mutants with mutations at serine 225; this residue was mutated to either alanine (S225A; phosphoablatant) or aspartic acid (S225D; phosphomimetic) in the context of both the JFH-1 cell culture infectious virus and a corresponding subgenomic replicon. The S225A mutant exhibited a 10-fold reduction in genome replication, whereas the S225D mutant replicated like the wild type. By confocal microscopy, we show that, in the case of the S225A mutant, the replication phenotype correlated with an altered subcellular distribution of NS5A. This phenotype was shared by viruses with other mutations in the low-complexity sequence I (LCS I), namely, S229D, S232A, and S235D, but not by viruses with mutations that caused a comparable replication defect that mapped to domain II of NS5A (P315A, L321A). Together with other components of the genome replication complex (NS3, double-stranded RNA, and cellular lipids, including phosphatidylinositol 4-phosphate), the mutation in NS5A was restricted to a perinuclear region. This phenotype was not due to cell confluence or another environmental factor and could be partially transcomplemented by wild-type NS5A. We propose that serine phosphorylation within LCS I may regulate the assembly of an active genome replication complex

The effect of ultrasonic excitation on the electrical properties and microstructure of printed electronic conductive inks

Ultrasonic Additive Manufacturing (UAM) is an advanced manufacturing technique, which enables the embedding of electronic components and interconnections within solid aluminium structures, due to the low temperature encountered during material bonding. In this study, the effects of ultrasonic excitation, caused by the UAM process, on the electrical properties and the microstructure of thermally cured screen printed silver conductive inks were investigated. The electrical resistance and the dimensions of the samples were measured and compared before and after the ultrasonic excitation. The microstructure of excited and unexcited samples was examined using combined Focused Ion Beam and Scanning Electron Microscopy (FIB/SEM) and optical microscopy. The results showed an increase in the resistivity of the silver tracks after the ultrasonic excitation, which was correlated with a change in the microstructure: the size of the silver particles increased after the excitation, suggesting that inter-particle bonding has occurred. The study also highlighted issues with short circuiting between the conductive tracks and the aluminium substrate, which were attributed to the properties of the insulating layer and the inherent roughness of the UAM substrate. However, the reduction in conductivity and observed short circuiting were sufficiently small and rare, which leads to the conclusion that printed conductive tracks can function as interconnects in conjunction with UAM, for the fabrication of novel smart metal components

“Raising the curtain on the equality theatre”: A study of recruitment to first healthcare job post-qualification in the UK National Health Service

Background UK equality law and National Health Service (NHS) policy requires racial equality in job appointments and career opportunities. However, recent national workforce race equality standard (WRES) data show that nearly all NHS organisations in the UK are failing to appoint ethnically diverse candidates with equivalent training and qualifications as their white counterparts. This is problematic because workforce diversity is associated with improved patient outcomes and other benefits for staff and organisations. Aim To better understand the reasons behind underrepresentation of ethnically diverse candidates in first NHS healthcare jobs post-qualification and to identify any structural or systemic barriers to employment for such groups. Methods The study was informed by critical theory and the authors’ interdisciplinary perspectives as educators and researchers in the healthcare professions. Data collected from semi-structured face-to-face interviews with 12 nurse and physiotherapy recruiting managers from two NHS trusts in London were analysed using a healthcare workforce equity and diversity conceptual lens we developed from the literature. Using this lens, we devised questions to examine six dimensions of equity and diversity in the interview data from recruiting managers. Results Recruiting managers said they valued the benefits of an ethnically diverse workforce for patients and their unit/organisation. However, their adherence to organisational policies for recruitment and selection, which emphasise objectivity and standardisation, acted as constraints to recognising ethnicity as an important issue in recruitment and workforce diversity. Some recruiting managers sense that there are barriers for ethnically diverse candidates but lacked information about workforce diversity, systems for monitoring recruitment, or ways to engage with staff or candidates to talk about these issues. Without this information there was no apparent problem or reason to try alternative approaches. Conclusion These accounts from 12 recruiting managers give a ‘backstage’ view into the reasons behind ethnic inequalities in recruitment to first healthcare job in the UK NHS. Adherence to recruitment and selection policies, which aim to support equality through standardisation and anonymisation, appear to be limiting workforce diversity and creating barriers for ethnically diverse candidates to attain the jobs that they are trained and qualified for. The Healthcare Workforce Equity + Diversity Lens we have developed can help to ‘raise the curtain on the equality theatre’ and inform more inclusive approaches to recruitment such as contextualised recruitment or effective allyship between employers and universities

Additive manufacturing of physical assets by using ceramic multicomponent extra-terrestrial materials

Powder Bed Fusion (PBF) is a range of advanced manufacturing technologies that can fabricate three-dimensional assets directly from CAD data, on a successive layer-by-layer strategy by using thermal energy, typically from a laser source, to irradiate and fuse particles within a powder bed.The aim of this paper was to investigate the application of this advanced manufacturing technique to process ceramic multicomponent materials into 3D layered structures. The materials used matched those found on the Lunar and Martian surfaces. The indigenous extra-terrestrial Lunar and Martian materials could potentially be used for manufacturing physical assets onsite (i.e., off-world) on future planetary exploration missions and could cover a range of potential applications including: infrastructure, radiation shielding, thermal storage, etc.Two different simulants of the mineralogical and basic properties of Lunar and Martian indigenous materials were used for the purpose of this study and processed with commercially available laser additive manufacturing equipment. The results of the laser processing were investigated and quantified through mechanical hardness testing, optical and scanning electron microscopy, X-ray fluorescence spectroscopy, thermo-gravimetric analysis, spectrometry, and finally X-ray diffraction.The research resulted in the identification of a range of process parameters that resulted in the successful manufacture of three-dimensional components from Lunar and Martian ceramic multicomponent simulant materials. The feasibility of using thermal based additive manufacturing with multi-component ceramic materials has therefore been established, which represents a potential solution to off-world bulk structure manufacture for future human space exploration

Cessation of mass drug administration for lymphatic filariasis in Zanzibar in 2006: was transmission interrupted?

BACKGROUND: Lymphatic filariasis (LF) is targeted for elimination through annual mass drug administration (MDA) for 4-6 years. In 2006, Zanzibar stopped MDA against LF after five rounds of MDA revealed no microfilaraemic individuals during surveys at selected sentinel sites. We asked the question if LF transmission was truly interrupted in 2006 when MDA was stopped. METHODOLOGY/PRINCIPAL FINDINGS: In line with ongoing efforts to shrink the LF map, we performed the WHO recommended transmission assessment surveys (TAS) in January 2012 to verify the absence of LF transmission on the main Zanzibar islands of Unguja and Pemba. Altogether, 3275 children were tested on both islands and 89 were found to be CFA positive; 70 in Pemba and 19 in Unguja. The distribution of schools with positive children was heterogeneous with pronounced spatial variation on both islands. Based on the calculated TAS cut-offs of 18 and 20 CFA positive children for Pemba and Unguja respectively, we demonstrated that transmission was still ongoing in Pemba where the cut-off was exceeded. CONCLUSIONS: Our findings indicated ongoing transmission of LF on Pemba in 2012. Moreover, we presented evidence from previous studies that LF transmission was also active on Unguja shortly after stopping MDA in 2006. Based on these observations the government of Zanzibar decided to resume MDA against LF on both islands in 2013

Risk of hospitalisation or death in households with a case of COVID-19 in England: an analysis using the HOSTED dataset

Objective: To determine whether household contacts of confirmed cases of COVID-19 have an increased risk of hospitalisation or death. Methods: We used the HOSTED dataset of index cases of COVID-19 in England between June and November 2020, linked to Secondary Uses Service data on hospital episodes and Office for National Statistics’ mortality data. Multivariable logistic regression models of the odds of household contacts being hospitalised or dying within six weeks of an index case, adjusted for case type, age, sex and calendar month were calculated. Excess risk was determined by comparing the first six weeks after the index case with 6-12 weeks after the index case in a survival analysis framework. Results: Index cases were more likely to be hospitalised or die than either secondary cases or non-cases, having adjusted for age and sex. There was an increased risk of hospitalisation for non-cases (adjusted hazard ratio (aHR) 1.10 (95% CI 1.04, 1.16)) and of death (aHR 1.57 (95% CI 1.14, 2.16)) in the first six weeks after an index case, compared to 6-12 weeks after. Conclusion: Risks of hospitalisation and mortality are predictably higher in cases compared to non-cases. The short-term increase in risks for non-case contacts following diagnosis of the index case may suggest incomplete case ascertainment among contacts, although this was relatively small

Monitoring the hepatitis C epidemic in England and evaluating intervention scale-up using routinely collected data.

In England, 160 000 individuals were estimated to be chronically infected with hepatitis C virus (HCV) in 2005 and the burden of severe HCV-related liver disease has increased steadily for the past 15 years. Direct-acting antiviral treatments can clear infection in most patients, motivating HCV elimination targets. However, the current burden of HCV is unknown and new methods are required to monitor progress. We employed a Bayesian back-calculation approach, combining data on severe HCV-related liver disease and disease progression, to reconstruct historical HCV incidence and estimate current prevalence in England. We explicitly modelled infections occurring in people who inject drugs, the key risk group, allowing information on the size of this population and surveillance data on HCV prevalence to inform recent incidence. We estimated that there were 143 000 chronic infections in 2015 (95% credible interval 123 000-161 000), with 34% and 54% in those with recent and past injecting drug use, respectively. Following the planned scale-up of new treatments, chronic infections were predicted to fall to 113 400 (94 900-132 400) by the end of 2018 and to 89 500 (71 300-108 600) by the end of 2020. Numbers developing severe HCV-related liver disease were predicted to fall by at least 24% from 2015 to 2020. Thus, we describe a coherent framework to monitor progress using routinely collected data, which can be extended to incorporate additional data sources. Planned treatment scale-up is likely to achieve 2020 WHO targets for HCV morbidity, but substantial efforts will be required to ensure that HCV testing and patient engagement are sufficiently high

Eradication of two incursions of the Red Imported Fire Ant in Queensland, Australia

Of the five known incursions of the highly invasive Red Imported Fire Ant in Australia, two are regarded to have been eradicated. As treatment efforts continue, and the programme evolves and new tools become available, eradication is still considered to be feasible for the remaining Red Imported Fire Ant populations with long-term commitment and support

Increased uptake and new therapies are needed to avert rising hepatitis C-related end stage liver disease in England: modelling the predicted impact of treatment under different scenarios.

BACKGROUND & AIMS: Hepatitis C (HCV) related disease in England is predicted to rise, and it is unclear whether treatment at current levels will be able to avert this. The aim of this study was to estimate the number of people with chronic HCV infection in England that are treated and assess the impact and costs of increasing treatment uptake. METHODS: Numbers treated were estimated using national data sources for pegylated interferon supplied, dispensed, or purchased from 2006 to 2011. A back-calculation approach was used to project disease burden over the next 30 years and determine outcomes under various scenarios of treatment uptake. RESULTS: 5000 patients were estimated to have been treated in 2011 and 28,000 in total from 2006 to 2011; approximately 3.1% and 17% respectively of estimated chronic infections. Without treatment, incident cases of decompensated cirrhosis and hepatocellular carcinoma were predicted to increase until 2035 and reach 2290 cases per year. Treatment at current levels should reduce incidence by 600 cases per year, with a peak around 2030. Large increases in treatment are needed to halt the rise; and with more effective treatment the best case scenario predicts incidence of around 500 cases in 2030, although treatment uptake must still be increased considerably to achieve this. CONCLUSIONS: If the infected population is left untreated, the number of patients with severe HCV-related disease will continue to increase and represent a substantial future burden on healthcare resources. This can be mitigated by increasing treatment uptake, which will have the greatest impact if implemented quickly