Isolation of Human Photoreceptor Precursors via a Cell Surface Marker Panel from Stem Cell-derived Retinal Organoids and Fetal Retinae

Loss of photoreceptor cells due to retinal degeneration is one of the main causes of blindness in the developed world. Although there is currently no effective treatment, cell replacement therapy using stem-cell-derived photoreceptor cells may be a feasible future treatment option. In order to ensure safety and efficacy of this approach, robust cell isolation and purification protocols must be developed. To this end, we previously developed a biomarker panel for the isolation of mouse photoreceptor precursors from the developing mouse retina and mouse embryonic stem cell cultures. In the current study we applied this approach to the human pluripotent stem cell (hPSC) system, and identified novel biomarker combinations that can be leveraged for the isolation of human photoreceptors. Human retinal samples and hPSC-derived retinal organoid cultures were screened against 242 human monoclonal antibodies using a high through-put flow cytometry approach. We identified 46 biomarkers with significant expression levels in the human retina and hPSC differentiation cultures. Human retinal cell samples, either from fetal tissue or derived from embryonic and induced pluripotent stem cell cultures, were FAC-sorted using selected candidate biomarkers that showed expression in discrete cell populations. Enrichment for photoreceptors and exclusion of mitotically active cells was demonstrated by immunocytochemical analysis with photoreceptor-specific antibodies and Ki-67. We established a biomarker combination, which enables the robust purification of viable human photoreceptors from both human retinae and hPSC-derived organoid cultures. This article is protected by copyright. All rights reserved

Evaluation of a risk-stratification strategy to improve primary care for low back pain: the MATCH cluster randomised trial protocol

Background Despite numerous options for treating back pain and the increasing healthcare resources devoted to this problem, the prevalence and impact of back pain-related disability has not improved. It is now recognized that psychosocial factors, as well as physical factors, are important predictors of poor outcomes for back pain. A promising new approach that matches treatments to the physical and psychosocial obstacles to recovery, the STarT Back risk stratification approach, improved patients’ physical function while reducing costs of care in the United Kingdom (UK). This trial evaluates implementation of this strategy in a United States (US) healthcare setting. Methods Six large primary care clinics in an integrated healthcare system in Washington State were block-randomized, three to receive an intensive quality improvement intervention for back pain and three to serve as controls for secular trends. The intervention included 6 one-hour training sessions for physicians, 5 days of training for physical therapists, individualized and group coaching of clinicians, and integration of the STarT Back tool into the electronic health record. This prognostic tool uses 9 questions to categorize patients at low, medium or high risk of persistent disabling pain with recommendations about evidence-based treatment options appropriate for each subgroup. Patients at least 18 years of age, receiving primary care for non-specific low back pain, were invited to provide data 1–3 weeks after their primary care visit and follow-up data 2 months and 6 months (primary endpoint) later. The primary outcomes are back-related physical function and pain severity. Using an intention to treat approach, intervention effects on patient outcomes will be estimated by comparing mean changes at the 2 and 6 month follow-up between the pre- and post-implementation periods. The inclusion of control clinics permits adjustment for secular trends. Differences in change scores by intervention group and time period will be estimated using linear mixed models with random effects. Secondary outcomes include healthcare utilization and adherence to clinical guidelines. Discussion This trial will provide the first randomized trial evidence of the clinical effectiveness of implementing risk stratification with matched treatment options for low back pain in a United States health care delivery system

The Bivariate Normal Copula

We collect well known and less known facts about the bivariate normal distribution and translate them into copula language. In addition, we prove a very general formula for the bivariate normal copula, we compute Gini's gamma, and we provide improved bounds and approximations on the diagonal.Comment: 24 page

Creating a Patient Registry for the Parkview Vein Clinic

Background/Objective: The Parkview Vein Clinic provides individualized care to patients suffering from venous disease by providing assessment, education, treatment, ultrasound imaging, and surgical interventions all under one roof. The Vein Clinic has experienced a consistent increase in patient volume since opening in 2019, creating a need for tracking patient outcomes. Patient registries are useful tools for tracking high volumes of patients, assessing outcomes, and improving treatment guidelines. The main objective of this quality improvement project is to define the workflow for creating a patient registry for the Vein Clinic and determine which data points are feasible to collect. Methods: This is a retrospective chart review of patients with the diagnosis of “venous stasis ulcer” seen at the Vein Clinic from September 2019 to July 2022. A total of 84 data fields were collected on each patient, including information on demographics, medical history, ulcer descriptions, imaging, procedure information, and post-procedure follow-up. The Society for Venous Surgery Vascular Quality Initiative was used as a template for registry design, with the goal of merging the registry with the national database in the future. Results: Venous ulcer information, including number of healed ulcers, duration of ulcer, and largest diameter active ulcer, was not readily accessible within the chart and required expanded review find and quantify. All other categories were readily accessible in the chart. Conclusions: The data collected by the registry will be useful for future quality improvement purposes of the Vein Clinic. Creation of a structured reporting template in Epic would help facilitate the ease and accuracy of data extraction and help maintain the internal validity of the registry. More detailed follow-up assessments should be implemented to track patient outcomes, which could include use of the Venous Clinical Severity Score or a patient-reported outcomes assessment

Earthworks risk assessment on a heritage railway

The UK is home to a substantial number of heritage and tourist railways, which make a significant contribution to their local economies. They are mostly constructed on the routes of closed lines, and include large numbers of earthworks of uncertain construction and unknown strength. Recently, there have been earthwork collapses, most notably on the Gloucester and Warwickshire Railway during 2010 and 2011. The Office of Rail Regulation has also noted a number of safety incidents on heritage railways, all attributable to management failures. This paper describes an analysis of the Victorian earthworks on the Bo'ness and Kinneil Railway, a 8 km-long heritage railway in central Scotland. The analysis and risk prioritisation method used by Network Rail was found to be unsuitable for direct application to heritage railways, owing to the different operating context. A new system was therefore developed, removing some risk factors from the Network Rail approach, adding others, and modifying further ones. The new system was successfully applied, and the Bo'ness and Kinneil Railway earthworks were found to be generally stable and safe

Phenotypic Variability of Childhood Charcot-Marie-Tooth Disease

IMPORTANCE: Disease severity of childhood Charcot-Marie-Tooth disease (CMT) has not been extensively characterized, either within or between types of CMT to date. OBJECTIVE: To assess the variability of disease severity in a large cohort of children and adolescents with CMT. DESIGN, SETTING, AND PARTICIPANTS: A cross-sectional study was conducted among 520 children and adolescents aged 3 to 20 years at 8 universities and hospitals involved in the Inherited Neuropathies Consortium between August 6, 2009, and July 31, 2014, in Australia, Italy, the United Kingdom, and the United States. Data analysis was conducted from August 1, 2014, to December 1, 2015. MAIN OUTCOMES AND MEASURES: Scores on the Charcot-Marie-Tooth Disease Pediatric Scale (CMTPedS), a well-validated unidimensional clinical outcome measure to assess disease severity. This instrument includes 11 items assessing fine and gross motor function, sensation, and balance to produce a total score ranging from 0 (unaffected) to 44 (severely affected). RESULTS: Among the 520 participants (274 males) aged 3 to 20 years, CMT type 1A (CMT1A) was the most prevalent type (252 [48.5%]), followed by CMT2A (31 [6.0%]), CMT1B (15 [2.9%]), CMT4C (13 [2.5%]), and CMTX1 (10 [1.9%]). Disease severity ranged from 1 to 44 points on the CMTPedS (mean [SD], 21.5 [8.9]), with ankle dorsiflexion strength and functional hand dexterity test being most affected. Participants with CMT1B (mean [SD] CMTPedS score, 24.0 [7.4]), CMT2A (29.7 [7.1]), and CMT4C (29.8 [8.6]) were more severely affected than those with CMT1A (18.9 [7.7]) and CMTX1 (males: 15.3 [7.7]; females: 13.0 [3.6]) (P < .05). Scores on the CMTPedS tended to worsen principally during childhood (ages, 3-10 years) for participants with CMT4C and CMTX1 and predominantly during adolescence for those with CMT1B and CMT2A (ages, 11-20 years), while CMT1A worsened consistently throughout childhood and adolescence. For individual items, participants with CMT4C recorded more affected functional dexterity test scores than did those with all other types of CMT (P < .05). Participants with CMT1A and CMTX1 performed significantly better on the 9-hole peg test and balance test than did those with all other types of CMT (P < .05). Participants with CMT2A had the weakest grip strength (P < .05), while those with CMT2A and CMT4C exhibited the weakest ankle plantarflexion and dorsiflexion strength, as well as the lowest long jump and 6-minute walk test distances (P < .05). Multiple regression modeling identified increasing age (r = 0.356, β = 0.617, P < .001) height (r = 0.251, β = 0.309, P = .002), self-reported foot pain (r = 0.162, β = .114, P = .009), and self-reported hand weakness (r = 0.243, β = 0.203, P < .001) as independent predictors of disease severity. CONCLUSIONS AND RELEVANCE: These results highlight the phenotypic variability within CMT genotypes and mutation-specific manifestations between types. This study has identified distinct functional limitations and self-reported impairments to target in future therapeutic trials

What is the 'problem' that outreach work seeks to address and how might it be tackled? Seeking theory in a primary health prevention programme

&lt;b&gt;Background&lt;/b&gt; Preventive approaches to health are disproportionately accessed by the more affluent and recent health improvement policy advocates the use of targeted preventive primary care to reduce risk factors in poorer individuals and communities. Outreach has become part of the health service response. Outreach has a long history of engaging those who do not otherwise access services. It has, however, been described as eclectic in its purpose, clientele and mode of practice; its effectiveness is unproven. Using a primary prevention programme in the UK as a case, this paper addresses two research questions: what are the perceived problems of non-engagement that outreach aims to address; and, what specific mechanisms of outreach are hypothesised to tackle these.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Methods&lt;/b&gt; Drawing on a wider programme evaluation, the study undertook qualitative interviews with strategically selected health-care professionals. The analysis was thematically guided by the concept of 'candidacy' which theorises the dynamic process through which services and individuals negotiate appropriate service use.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Results&lt;/b&gt; The study identified seven types of engagement 'problem' and corresponding solutions. These 'problems' lie on a continuum of complexity in terms of the challenges they present to primary care. Reasons for non-engagement are congruent with the concept of 'candidacy' but point to ways in which it can be expanded.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Conclusions&lt;/b&gt; The paper draws conclusions about the role of outreach in contributing to the implementation of inequalities focused primary prevention and identifies further research needed in the theoretical development of both outreach as an approach and candidacy as a conceptual framework

Predictors and temporal trend of flu vaccination in auto-immune rheumatic diseases in the UK: a nationwide prospective cohort study

Objectives: To [1] examine temporal trend in uptake of seasonal influenza vaccine (SIV) in the UK, [2] explore disease and demographic factors associated with vaccination. Methods: 32,751 people with auto-immune rheumatic diseases (AIRDs) prescribed disease modifying anti-rheumatic drugs (DMARDs) between 2006 and 2016 were identified from the Clinical Practice Research Datalink. The proportion vaccinated between 01/September of one year and 31/March of next year was calculated and stratified by age, other indications for vaccination, AIRD type, and number of DMARDs prescribed. Stata and Joinpoint regression programs were used. Results: SIV uptake was high in those aged ≥65 years (82.3% and 80.7% in 2006-07 and 2015-16 respectively). It was significantly lower in other age groups, but improved over time with 51.9% and 61.9% in the 45-64 year age group, and 32.3% and 50.1% in the <45 year age group being vaccinated in 2006-07 and 2015-16 respectively. While 64.9% of the vaccinations in those ≥65 years old occurred by 3rd November, in time to mount a protective immune response before the influenza activity becomes substantial in UK, only 38.9% in the 45-64 year and 26.2% in the <45 year age group without any other reason for vaccination received SIV by this date. Women, those with additional indications for vaccination, on multiple DMARDs and with SLE were more likely to be vaccinated. Conclusion: SIV uptake is low in the under 65s, and the majority of them are not vaccinated in time. Additional effort is required to promote timely uptake of SIV in this population

Caffeinated energy drinks and effects in UK young people

International systematic review evidence indicates an association of caffeinated energy drink use with physical symptoms and lifestyle but is unclear about associations with mental health and behavioural outcomes. The design of studies included in the reviews and the quality of the systematic reviews themselves limits the strength of the conclusions. The lack of UK research in the reviews prompted our analysis of UK population-level data. Our analysis of UK data suggests that many children in the UK consume CEDs with higher consumption reported by older children, by boys, and by those living in northern areas or in more deprived regions. Findings also suggest associations between consumption and physical, psychological, social and educational symptoms, behaviours and wellbeing. A lack of studies that measure these variables over time meant we were unable to determine whether CED consumption is the cause of associated symptoms, behaviours and wellbeing. Future research on this topic should employ longitudinal methods to examine whether CED consumption is responsible for poorer health and wellbeing. Research should also examine the influence of geographic region and deprivation on children’s caffeinated energy drink consumption. The measurement of caffeinated energy drink consumption in the future needs to be consistent across surveys, so that a clearer picture of the frequency, timing and dosage can be established. UK-wide exploration of the context and reasons for caffeinated energy drink consumption is needed and should include examining consumption of CEDs with alcohol in older children aged 16 to 17 years