KEY MESSAGE : A switchgrass protoplast system was developed, achieving a cost reduction of ~1000-fold, a threefold increase in transformation efficiency, and a fourfold reduction in required DNA ...quantity compared to previous methods. In recent years, there has been a resurgence in the use of protoplast systems for rapid screening of gene silencing and genome-editing targets for siRNA, miRNA, and CRISPR technologies. In the case of switchgrass (Panicum virgatum L.), to achieve economic feasibility for biofuel production, it is necessary to develop plants with decreased cell wall recalcitrance to reduce processing costs. To achieve this goal, transgenic plants have been generated with altered cell wall chemistry; however, with limited success owing to the complexity of cell walls. Because of the considerable cost, time, and effort required to screen transgenic plants, a protoplast system that can provide data at an early stage has potential to eliminate low performing candidate genes/targets prior to the creation of transgenic plants. Despite the advantages of protoplast systems, protoplast isolation in switchgrass has proven costly, requiring expensive lab-grade enzymes and high DNA quantities. In this paper, we describe a low-cost protoplast isolation system using a mesophyll culture approach and a cell suspension culture. Results from this work show a cost reduction of ~1000-fold compared to previous methods of protoplast isolation in switchgrass, with a cost of $0.003 (USD) per reaction for mesophyll protoplasts and $0.018 for axenic cell culture-derived protoplasts. Further, the efficiency of protoplast transformation was optimized threefold over previous methods, despite a fourfold reduction in DNA quantity. The methods developed in this work remove the cost barrier previously limiting high-throughput screening of genome-editing and gene silencing targets in switchgrass, paving the way for more efficient development of transgenic plants.
The quality of life of people receiving health and social care is an important indicator of service quality, but the relationship between patient experience and outcomes and regulator quality ratings ...in England is unknown. In 2013, the health and social care regulator in England, the Care Quality Commission (CQC), introduced a new ratings system and by February 2017, all social care services were inspected and awarded new quality ratings (outstanding, good, requires improvement and inadequate). This study aimed to explore whether quality ratings were associated with residents' quality of life, controlling for confounding variables.
We conducted a nested, cross-sectional study, collecting social care-related quality of life (SCRQoL) data for 293 older care home residents in 34 care homes (20 nursing and 14 residential) in the South East of England. CQC ratings and other resident and home-level variables were also collected for the analysis. Multilevel modelling explored whether residents' social care-related quality of life (SCRQoL) was associated with regulator ratings, controlling for confounding variables.
Outstanding and good homes were collapsed into one category and compared with homes requiring improvement. Nationally, only 2 % of care homes for older people are rated as inadequate and it was not possible to capture sufficient numbers for the analysis. We recruited one but it was re-inspected during the fieldwork period and its rating changed to requires improvement. The random intercept multilevel model, which accounted for 16.93% of the differences in SCRQoL within homes and 69.80% between, indicated that better SCRQoL was significantly associated with being female, better functioning, no dementia diagnosis, fewer communication difficulties, and living in a care home rated as outstanding/good by CQC. Size of home and registration category were not significant predictors.
This study found evidence that quality ratings are associated with residents' SCRQoL. As well as aiming to improve quality and ensure minimum standards, quality ratings have the potential to inform user choice and help the public compare care homes based on quality. Future research to establish the generalisability and replicability of the results is required.
Social care-related quality of life is a key outcome indicator used in the evaluation of social care interventions and policy. It is not, however, always possible to collect quality of life data by ...self-report even with adaptations for people with cognitive or communication impairments. A new proxy-report version of the Adult Social Care Outcomes Toolkit (ASCOT) measure of social care-related quality of life was developed to address the issues of wider inclusion of people with cognitive or communication difficulties who may otherwise be systematically excluded. The development of the proxy-report ASCOT questionnaire was informed by literature review and earlier work that identified the key issues and challenges associated with proxy-reported outcomes.
To evaluate the acceptability and content validity of the ASCOT-Proxy, qualitative cognitive interviews were conducted with unpaid carers or care workers of people with cognitive or communication impairments. The proxy respondents were invited to 'think aloud' while completing the questionnaire. Follow-up probes were asked to elicit further detail of the respondent's comprehension of the format, layout and content of each item and also how they weighed up the options to formulate a response.
A total of 25 unpaid carers and care workers participated in three iterative rounds of cognitive interviews. The findings indicate that the items were well-understood and the concepts were consistent with the item definitions for the standard self-completion version of ASCOT with minor modifications to the draft ASCOT-Proxy. The ASCOT-Proxy allows respondents to rate the proxy-proxy and proxy-patient perspectives, which improved the acceptability of proxy report.
A new proxy-report version of ASCOT was developed with evidence of its qualitative content validity and acceptability. The ASCOT-Proxy is ready for empirical testing of its suitability for data collection as a self-completion and/or interview questionnaire, and also evaluation of its psychometric properties.
Outcomes-based policy and administration of public services present a compelling argument for the value of outcomes data. However, there are a number of challenges inherent in collecting these data ...from people who are unable to complete a paper-based survey or interview due to cognitive or communication impairments. In this paper, we explore the views of being a proxy from the perspective of unpaid carers and paid carers who may be asked to act as a proxy on behalf of the person(s) they care for. We consider the key issues that need to be addressed when adapting an instrument designed to measure social care outcomes, the Adult Social Care Outcomes Tool (ASCOT), into a proxy-report tool.
Participants took part in either a focus group (35 paid carers in eight focus groups), or a one-to-one interview (eight unpaid carers). All participants were recruited via carer organisations and care providers. Transcripts, field notes and audio data collected during focus groups and interviews were analysed using a thematic framework approach.
Participants agreed that any person acting as a proxy would need to be very familiar with the care recipient, as well as their needs and care provision. A number of provisions for proxy respondents were proposed to improve face validity and acceptability of completing a questionnaire by proxy, and to ensure that any potential bias is reduced in the design of the questionnaire. These included: providing two sets of response options for each proxy perspective (the proxy themselves and the proxy view of how they think the care recipient would respond); a comments box to help people explain why they have selected a given response option (especially where these indicate unmet need); and providing clear guidance for the proxy respondent on how they should complete the questionnaire.
This study has shown some of the challenges involved in assessing outcomes by proxy and explored some potential ways these can be mitigated. The findings highlight the benefits of developing and testing proxy measures in a robust way to widen participation in social care research.
Introductionsyphilis and its outcomes remain a healthcare system burden with adverse consequences such as stillbirths, neonatal deaths and spontaneous abortions among others. The situation might have ...worsened because the COVID-19 pandemic has caused a major attention drift from other diseases. Additionally, much as testing for syphilis is a routine practice among pregnant mothers, its proportion is not known in urban health care setting. A study to determine the prevalence of syphilis among pregnant mothers in an urban poor setting is warranted.Methodsa cross-sectional study was conducted among pregnant women who attended antenatal care at Kawaala Health Centre IV in Kampala Capital City between December 2019 to March 2020. Informed consent was sought from study participants prior to data collection using structured questionnaires. Whole blood was collected and tested using SD Bioline HIV/syphilis duo rapid test kit (SD Standard Diagnostics, INC, Korea). Data analysis was done using STATA 14.2.Resultsone thousand one hundred and sixty-nine pregnant women participated in the study, with a mean age of 25 years. About 27% of them had completed only primary-level education. Approximately 6% of the participants were HIV seropositive. The prevalence of syphilis was 5.9% (69/1169). HIV positivity (aOR: 4.13, 95%CI: 2.05-8.34), elevated blood pressure (aOR: 2.84, 95%CI: 1.42-5.69), and status of previous pregnancy (aOR: 0.21, 95%CI: 0.05-0.89) were significant predictors of the risk of syphilis among pregnant women in this setting.Conclusionthe prevalence of syphilis among pregnant women in urban poor settings is not low and so must not be underestimated. The potential drivers of syphilis among pregnant women are HIV, elevated blood pressure, and status of previous pregnancy. There should be increased awareness about routine syphilis testing among pregnant mothers attending antenatal care.
A 25-year-old woman with a background history of bronchial asthma and intellectual disability presented to hospital with progressively worsening dyspnoea. Despite testing negative four times for ...coronavirus disease infection by nasopharyngeal swab reverse-transcriptase polymerase chain reaction, her clinical symptoms of hypoxaemic respiratory failure and radiological findings on computed tomography pulmonary angiogram were consistent with coronavirus disease pneumonia. Although she made a quick recovery in the intensive care unit with a combination of empirical antibiotics, corticosteroids, high flow nasal oxygen, therapeutic anticoagulation and awake semi proning, her protracted hospital course due to persistent sinus tachycardia remained challenging. A diagnosis of potential postural orthostatic tachycardia syndrome was explored during the acute phase of illness following an active stand test and exclusion of other causes. She was treated with beta blockers as she failed to improve with non-pharmacological measures. We searched for similar cases by analysing the literature databases. Our case aims to stress the importance of recognising and treating patients with negative nasal reverse-transcriptase polymerase chain reaction swabs as coronavirus disease infection, especially if there is strong evidence of clinical and radiological findings where diagnosis is often under recognised in asthmatics with intellectual disability.
Context: Social care outcomes (the effect of services on the quality of life of people with support needs and unpaid carers) have been proposed as a way of improving the quality and effectiveness of ...care. Outcomes have also been proposed as a way of reconceptualising ‘needs’ that have applications in needs assessment, care planning, evaluation and care practice. Objectives: The study aimed to provide insights into social care professionals’ experiences and views on the collection and application of outcomes data in practice and what they believe are the benefits, challenges and barriers to implementation. Methods: Interviews were conducted with 25 social care professionals in England and analysed using a framework approach. Findings: Participants reported perceived benefits of using outcomes data, especially to focus effort on improving the well-being of people with support needs and carers. Perceived challenges include requirements for data collection set by funders/commissioners, the volume of data collected, difficulties in separating non-service-related influences on outcomes and the format of collection. Participants felt a more flexible approach might facilitate more meaningful conversations, rather than a ‘tick-box’ exercise. Limitations: The study sample was purposive, based on established connections. It only included professionals from London and South East and Central England. Implications: Although outcomes are perceived as important in helping to improve people’s lives, social care professionals identified a number of challenges. Further research to understand and address these challenges is needed.
ObjectiveLiving Library events involve people being trained as living ‘Books’, who then discuss aspects of their personal experiences in direct conversation with attendees, referred to as ‘Readers’. ...This study sought to generate a realist programme theory and a theory-informed implementation guide for a Library of Lived Experience for Mental Health (LoLEM).DesignIntegrated realist synthesis and experience-based co-design.SettingTen online workshops with participants based in the North of England.ParticipantsThirty-one participants with a combination of personal experience of using mental health services, caring for someone with mental health difficulties and/or working in mental health support roles.ResultsDatabase searches identified 30 published and grey literature evidence sources which were integrated with data from 10 online co-design workshops conducted over 12 months. The analysis generated a programme theory comprising five context-mechanism-outcome (CMO) configurations. Findings highlight how establishing psychological safety is foundational to productive Living Library events (CMO 1). For Readers, direct conversations humanise others’ experiences (CMO 2) and provide the opportunity to flexibly explore new ways of living (CMO 3). Through participation in a Living Library, Books may experience personal empowerment (CMO 4), while the process of self-authoring and co-editing their story (CMO 5) can contribute to personal development. This programme theory informed the co-design of an implementation guide highlighting the importance of tailoring event design and participant support to the contexts in which LoLEM events are held.ConclusionsThe LoLEM has appeal across stakeholder groups and can be applied flexibly in a range of mental health-related settings. Implementation and evaluation are required to better understand the positive and negative impacts on Books and Readers.Trial registration numberPROSPERO CRD42022312789.
IntroductionPeople with lived expertise in managing mental health challenges can be an important source of knowledge and support for other people facing similar challenges, and for carers to learn ...how best to help. However, opportunities for sharing lived expertise are limited. Living libraries support people with lived expertise to be ‘living books’, sharing their experiences in dialogue with ‘readers’ who can ask questions. Living libraries have been piloted worldwide in health-related contexts but without a clear model of how they work or rigorous evaluation of their impacts. We aim to develop a programme theory about how a living library could be used to improve mental health outcomes, using this theory to codesign an implementation guide that can be evaluated across different contexts.Methods and analysisWe will use a novel integration of realist synthesis and experience-based codesign (EBCD) to produce a programme theory about how living libraries work and a theory and experience informed guide to establishing a library of lived experience for mental health (LoLEM). Two workstreams will run concurrently: (1) a realist synthesis of literature on living libraries, combined with stakeholder interviews, will produce several programme theories; theories will be developed collaboratively with an expert advisory group of stakeholders who have hosted or taken part in a living library and will form our initial analysis framework; a systematic search will identify literature about living libraries; data will be coded into our analysis framework, and we will use retroductive reasoning to explain living libraries’ impacts across multiple contexts. Individual stakeholder interviews will help refine and test theories; (2) data from workstream 1 will inform 10 EBCD workshops with people with experience of managing mental health difficulties and health professionals to produce a LoLEM implementation guide; data from this process will also inform the theory in workstream 1.Ethics and disseminationEthical approval was granted by Coventry and Warwick National Health Service Research Ethics Committee on 29 December 2021 (reference number 305975). The programme theory and implementation guide will be published as open access and shared widely through a knowledge exchange event, a study website, mental health provider and peer support networks, peer reviewed journals and a funders report.PROSPERO registration detailsCRD42022312789.
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