Efficient clinical-grade γ-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing.

γ-Retroviral vectors (γ-RV) are powerful tools for gene therapy applications. Current clinical vectors are produced from stable producer cell lines which require minimal further downstream processing, while purification schemes for γ-RV produced by transient transfection have not been thoroughly investigated. We aimed to develop a method to purify transiently produced γ-RV for early clinical studies. Here, we report a simple one-step purification method by high-speed centrifugation for γ-RV produced by transient transfection for clinical application. High-speed centrifugation enabled the concentration of viral titers in the range of 107-108 TU/mL with >80% overall recovery. Analysis of research-grade concentrated vector revealed sufficient reduction in product- and process-related impurities. Furthermore, product characterization of clinical-grade γ-RV by BioReliance demonstrated two-logs lower impurities per transducing unit compared with regulatory authority-approved stable producer cell line vector for clinical application. In terms of CAR T cell manufacturing, clinical-grade γ-RV produced by transient transfection and purified by high-speed centrifugation was similar to γ-RV produced from a clinical-grade stable producer cell line. This method will be of value for studies using γ-RV to bridge vector supply between early- and late-stage clinical trials.

SR-BI deficiency disassociates obesity from hepatic steatosis and glucose intolerance development in high fat diet-fed mice.

Scavenger receptor BI (SR-BI) has been suggested to modulate adipocyte function. To uncover the potential relevance of SR-BI for the development of obesity and associated metabolic complications, we compared the metabolic phenotype of wild-type and SR-BI deficient mice fed an obesogenic diet enriched in fat. Both male and female SR-BI knockout mice gained significantly more weight as compared to their wild-type counterparts in response to 12 weeks high fat diet feeding (1.5-fold; P < .01 for genotype). Plasma free cholesterol levels were ~2-fold higher (P < .001) in SR-BI knockout mice of both genders, whilst plasma cholesteryl ester and triglyceride concentrations were only significantly elevated in males. Strikingly, the exacerbated obesity in SR-BI knockout mice was paralleled by a better glucose handling. In contrast, only SR-BI knockout mice developed atherosclerotic lesions in the aortic root, with a higher predisposition in females. Biochemical and histological studies in male mice revealed that SR-BI deficiency was associated with a reduced hepatic steatosis degree as evident from the 29% lower (P < .05) liver triglyceride levels. Relative mRNA expression levels of the glucose uptake transporter GLUT4 were increased (+47%; P < .05), whilst expression levels of the metabolic PPARgamma target genes CD36, HSL, ADIPOQ and ATGL were reduced 39%-58% (P < .01) in the context of unchanged PPARgamma expression levels in SR-BI knockout gonadal white adipose tissue. In conclusion, we have shown that SR-BI deficiency is associated with a decrease in adipocyte PPARgamma activity and a concomitant uncoupling of obesity development from hepatic steatosis and glucose intolerance development in high fat diet-fed mice.

Socioeconomic disparities in orthodontic treatment outcomes and expenditure on orthodontics in England's state-funded National Health Service: a retrospective observational study.

BACKGROUND: This study aimed to assess whether there are potential areas for efficiency improvements in the National Health Service (NHS) orthodontic service in North West England and to assess the socioeconomic status (SES)-related equity of the outcomes achieved by the NHS. METHODS: The study involved a retrospective analysis of 2008-2012 administrative data, and the study population comprised patients aged ≥10 who started NHS primary care orthodontic treatment in North West England in 2008. The proportions of treatments that were discontinued early and ended with residual need (based on post-treatment Index of Orthodontic Treatment Need [IOTN] scores that met or exceeded the NHS eligibility threshold of 3.6) and the associated NHS expenditure were calculated. In addition, the associations with SES were investigated using linear probability models. RESULTS: We found that 7.6% of treatments resulted in discontinuation (which was associated with an NHS annual expenditure of £2.3 m), and a further 19.4% (£5.9 m) had a missing outcome record. Furthermore, 5.2% of treatments resulted in residual need (£1.6 m), and a further 38.3% (£11.6 m) had missing IOTN data (due to either a missing outcome record or an incomplete IOTN outcome field in the record), which led to an annual NHS expenditure of £13.2 m (44% of the total expenditure) on treatments that are a potential source of inefficiency. Compared to the patients in the highest SES group, those in the lower SES groups were more likely both to discontinue treatment and to have residual need on treatment completion. CONCLUSIONS: Substantial inefficiencies were evident in the NHS orthodontic service, with 7.6% of treatments ending in discontinuation (£2.3 m) and 5.2% ending with residual need (£1.6 m). Over a third of cases had unreported IOTN outcome scores, which highlights the need to improve the outcome monitoring systems. In addition, the SES gradients indicate inequity in the orthodontic outcomes, with children from disadvantaged communities having poorer outcomes compared to their more affluent peers.

Case 11: large, infected, necrotic mixed-aetiology leg ulcer.

This painful and malodorous ulcer was covered with 40% necrotic tissue. Combined use of octenilin Wound gel and Wound Irrigation Solution gently debrided the necrotic tissue, and helped reduce the pain and malodour. Within 6 weeks, the wound was covered with granulation tissue.

Case 12: a neuroischaemic ulcer on the heel.

This necrotic ulcer was managed with sharp debridement, octenilin Wound Irrigation Solution and Wound Gel, plus a non-adherent dressing. The significant oedema in the lower limb was controlled with reduced compression. The wound improved steadily over time, healing on week 16.

Case 13: chronic painful ulcer on the heel of a diabetic foot.

This painful ulcer, which had signs of biofilm, was covered in necrotic tissue. Octenilin Wound Gel was used to soften the necrosis in preparation for sharp debridement. Three weeks later, the devitalised tissue had been completely removed, while there was a 50% reduction in wound size at week 5.

The effect of different methods of remuneration on the behaviour of primary care dentists.

BACKGROUND: Methods of remuneration have been linked with the professional behaviour of primary care physicians. In dentistry, this can be exacerbated as clinicians operate their practices as businesses and take the full financial risk of the provision of services. The main methods for remunerating primary care dentists include fee-for-service, fixed salary and capitation payments. The aim of this review was to determine the impact that these remuneration mechanisms have upon primary care dentists' behaviour. OBJECTIVES: To evaluate the effects of different methods of remuneration on the level and mix of activities provided by primary care dentists and the impact this has on patient outcomes. SEARCH METHODS: We searched the Cochrane Effective Practice and Organisation of Care (EPOC) Group Specialised Register; the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 7, 2013); MEDLINE (Ovid) (1947 to 11 June 2013); EMBASE (Ovid) (1947 to 11 June 2013); EconLit (1969 to 11 June 2013); the NHS Economic Evaluation Database (EED) (11 June 2013); and the Health Economic Evaluations Database (HEED) (11 June 2013). We conducted cited reference searches for the included studies in ISI Web of Knowledge; searched grey literature sources; handsearched selected journals; and contacted authors of relevant studies. SELECTION CRITERIA: Primary care dentists were defined as clinicians that deliver routine or mainstream dental care in a primary care environment. We included randomised controlled trials (RCTs), non-randomised controlled clinical trials (NRCTs), controlled before-after (CBA) studies and interrupted time series (ITS) studies. The methods of remuneration that we considered were: fee-for-service, fixed salary and capitation payments. Primary outcome measures were: measures of clinical activity; volume of clinical activity undertaken; time taken and clinical session length, or both; clinician type utilised; measures of health service utilisation; access and attendance as a proportion of the population; re-attendance rates; recall frequency; levels of oral health inequalities; non-attendance rates; healthcare costs; measures of patient outcomes; disease reduction; health maintenance; and patient satisfaction. We also considered measures of practice profitability/income and any reported unintended effects of the included methods of remuneration. DATA COLLECTION AND ANALYSIS: Three of the review authors (PRB, JP, AMG) independently reviewed titles and abstracts and resolved disagreements by discussion. The same three review authors undertook data extraction and assessed the quality of the evidence from all the studies that met the selection criteria, according to Cochrane Collaboration procedures. MAIN RESULTS: Two cluster-RCTs, with data from 503 dental practices, representing 821 dentists and 4771 patients, met the selection criteria. We judged the risk of bias to be high for both studies and the overall quality of the evidence was low/very low for all outcomes, as assessed using the GRADE approach.One study used a factorial design to investigate the impact of fee-for-service and an educational intervention on the placement of fissure sealants in permanent molar teeth. The authors reported a statistically significant increase in clinical activity in the arm that was incentivised with a fee-for-service payment. However, the study was conducted in the four most deprived areas of Scotland, so the applicability of the findings to other settings may be limited. The study did not report data on measures of health service utilisation or measures of patient outcomes.The second study used a parallel group design undertaken over a three-year period to compare the impact of capitation payments with fee-for-service payments on primary care dentists' clinical activity. The study reported on measures of clinical activity (mean percentage of children receiving active preventive advice, health service utilisation (mean number of visits), patient outcomes (mean number of filled teeth, mean percentage of children having one or more teeth extracted and the mean number of decayed teeth) and healthcare costs (mean expenditure). Teeth were restored at a later stage in the disease process in the capitation system and the clinicians tended to see their patients less frequently and tended to carry out fewer fillings and extractions, but also tended to give more preventive advice.There was insufficient information regarding the cost-effectiveness of the different remuneration methods. AUTHORS' CONCLUSIONS: Financial incentives within remuneration systems may produce changes to clinical activity undertaken by primary care dentists. However, the number of included studies is limited and the quality of the evidence from the two included studies was low/very low for all outcomes. Further experimental research in this area is highly recommended given the potential impact of financial incentives on clinical activity, and particular attention should be paid to the impact this has on patient outcomes.

Achieving consensus in pressure ulcer reporting.

Pressure ulcers are considered to be a key quality indicator and healthcare providers in England are required to report local pressure ulcer rates. However, there is a lack of standardisation in reporting due to lack of national guidance. The Tissue Viability Society has sought to develop consensus amongst all concerned parties on the most useful and robust methods of data collection. This document has been developed following a consensus meeting and consultation with the majority of Tissue Viability Nurses across England and provides guidance on reporting pressure ulcer rates. It is intended for use all organisations that are involved in the reporting of pressure ulcers. It represents the consensus view of a large number of Tissue Viability Nurses from across England and we recommend its adoption.