HUMAN CELLS FOR PROSTATE CANCER VACCINE THERAPY - THE IMPACT OF CENTRIFUGATION UPON KEY PRODUCT QUALITY ATTRIBUTES

Centrifugation is a unit operation used within a wide range of bioprocesses, including the production of a whole cell cancer vaccine to treat Hormone Refractory Prostate Cancer (HRPC) which has been tested in Phase II clinical trials. To quantify the effect of centrifugation-related stresses upon a whole cell vaccine population, a design of experiments (DOE) based investigation at micro-scale was implemented. Qualification of the cells may be by membrane integrity and by surface marker density as well as cytokines released and retained during bioprocessing. These and possibly other factors combine to affect the biopotency of the cells and their clinical efficacy. A range of processing factors that included relative centrifugal force, spin time and ambient cell holding time prior to processing were investigated, and their impact upon an array of cell quality attributes measured. A screening study indicated that both relative centrifugal force (RCF) and spin time were statistically significant factors with regards to the loss of cell membrane integrity. A range of two or more factor interactions were also suggested as having a significant negative impact upon the key cell quality attribute of membrane integrity, illustrating the power of multi-factorial experimental design. Data also indicated that loss of cell size was as the result of an increase within all three processing parameters, resulting in the emergence of a smaller, membrane-compromised cell population. Cell surface phenotype analysis by quantitative flow cytometry suggested no significant change in the surface staining profile for the range of product quality surface markers tested. The findings should allow for the creation of a design space for associated centrifugation operations, allowing an optimum processing window to be established. A UK Technology Strategy Board funded program in collaboration with LGC, Nottingham Trent University (bioinformation centre) and formerly with Onyvax

ULTRA-SCALE DOWN STUDIES OF HUMAN CELL BIOPROCESSING FOR A PROSTATE CANCER VACCINE THERAPY - THE IMPACT OF CAPILLARY SHEAR

The scale-up and manufacturing of therapies based on intact whole cells presents a major challenge for development scientists and engineers due to the stress-reactive nature of these cells. The administrated cells may be characterized in terms of their membrane integrity and their surface markers and eventually their biopotency. The challenge is to process the cells at various scales and in a way which maintains these cell properties. Also during formulation the presence of cytokines produced by cells prior to their inactivation is a critical factor. This poster presents an approach to allow the rapid characterization of human cell lines in terms of their resistance to hydrodynamic stress. An ultra scale-down method has been developed which allows investigation with small quantities of cells commonly available at the early discovery stage. The study describes controlled flow through a capillary device where cells are exposed to several defined hydrodynamic stresses. A Design of Experiments approach was used to understand the combined effect of many process parameters such as flow rate, length of capillary and number of passes. This was followed by an additional set of detailed ultra scale-down experiments where other critical quality attributes like cell size, surface phenotype, biopotency and cytokine release were measured. Computational fluid dynamics was used to describe the capillary entry region which allows the cells to be characterized in terms of a critical stress below which there is no significant damage to cell integrity or surface phenotype. A UK Technology Strategy Board funded program in collaboration with LGC, Nottingham Trent University and originally with Onyvax Ltd

Process development for recovery of crystals using DoE

Process design of a dead end filtration and resulting performance at industrial scale relies on small-scale data acquired with a few mL of suspended particles in a filter with a diameter of a few mm. and milliliter scale filtration systems. For rapid process development, it is important to gain early information on the filterability of the process stream regarding suspension flowability and cake compaction under different process conditions such as pressure difference, and filter pore size as a function of particle size distribution (PSD) and crystal concentration. The effect of these process variables was therefore investigated on process performance and product yield. A design of experiment (DoE) tool was also employed to identify optimum process conditions and to create a predictive model. In this work, a case study is presented on the characterisation of an active pharmaceutical ingredient (API) - paracetamol using laser diffraction and microscopic imaging techniques. The effect of changes in process parameters including pressure difference, PSD and concentration on filterability were investigated experimentally.The filtration rate was investigated at pressure driving forces of 100 to 700 mbar and for mean particle sizes ranging from micronised to granulated. The experimental design allowed the utilisation of filter capacity to be optimised and also enables predictive assessment of other process conditions. Validation of predictive model using parity plots shows acceptable agreement with regression >90% between the predicted and experimental data. The findings enhance understanding of the filtration step resulting in robust process development at laboratory scale

COSMOS dataset for co-existence/ interference analysis and simultaneous scene representation by automotive radar and video with GPS/IMU ground truth - Sub-Dataset C

The objectives of the described radar trials were to conduct radar measurements at the background of interference in the 76 – 81 GHz frequency band to: • Estimate the impact of interference in an adaptive cruise control (ACC) and cross-traffic alert (CTA) scenarios. • Analyse radar field of view shadowing due to a close target. • Identify an oncoming vehicle from the received interference which is otherwise blind due to radar field of view (FoV) obstruction. • Estimate the multipath interference in a reflective scenario. Repository Overview: The full dataset collected during the trails is over 2 TB. Depending upon the scenario and data collection duration, the size of raw data captured from INRAS Radarlog varies from 1 GB to 7 GB whereas for INRAS Radarbook, it varies from 1 GB to 4 GB. Therefore, due to extremely large files sizes, only the most suitable representative of the defined use-cases is included in the repository. Moreover, the full post-processed radar imagery is only shown for a few example cases. Additional data may be available on request

Evaluating the effectiveness and cost effectiveness of the 'strengthening families, strengthening communities' group-based parenting programme: study protocol and initial insights.

Funder: Public Health Research Programme; Grant(s): NIHR-PHR: 16/122/35BACKGROUND: Up to 20% of UK children experience socio-emotional difficulties which can have serious implications for themselves, their families and society. Stark socioeconomic and ethnic inequalities in children's well-being exist. Supporting parents to develop effective parenting skills is an important preventive strategy in reducing inequalities. Parenting interventions have been developed, which aim to reduce the severity and impact of these difficulties. However, most parenting interventions in the UK focus on early childhood (0-10 years) and often fail to engage families from ethnic minority groups and those living in poverty. Strengthening Families, Strengthening Communities (SFSC) is a parenting programme designed by the Race Equality Foundation, which aims to address this gap. Evidence from preliminary studies is encouraging, but no randomised controlled trials have been undertaken so far. METHODS/DESIGN: The TOGETHER study is a multi-centre, waiting list controlled, randomised trial, which aims to test the effectiveness of SFSC in families with children aged 3-18 across seven urban areas in England with ethnically and socially diverse populations. The primary outcome is parental mental well-being (assessed by the Warwick-Edinburgh Mental Well-Being Scale). Secondary outcomes include child socio-emotional well-being, parenting practices, family relationships, self-efficacy, quality of life, and community engagement. Outcomes are assessed at baseline, post intervention, three- and six-months post intervention. Cost effectiveness will be estimated using a cost-utility analysis and cost-consequences analysis. The study is conducted in two stages. Stage 1 comprised a 6-month internal pilot to determine the feasibility of the trial. A set of progression criteria were developed to determine whether the stage 2 main trial should proceed. An embedded process evaluation will assess the fidelity and acceptability of the intervention. DISCUSSION: In this paper we provide details of the study protocol for this trial. We also describe challenges to implementing the protocol and how these were addressed. Once completed, if beneficial effects on both parental and child outcomes are found, the impact, both immediate and longer term, are potentially significant. As the intervention focuses on supporting families living in poverty and those from minority ethnic communities, the intervention should also ultimately have a beneficial impact on reducing health inequalities. TRIAL REGISTRATION: Prospectively registered Randomised Controlled Trial ISRCTN15194500

Human occupation of northern Europe in MIS 13: Happisburgh Site 1 (Norfolk, UK) and its European context

The timing, environmental setting and archaeological signatures of an early human presence in northernEurope have been longstanding themes of Palaeolithic research. In the space of 20 years, the earliestrecord of human occupation in Britain has been pushed back from 500 ka (Boxgrove) to 700 ka (Pakefield)and then to >800 ka (Happisburgh Site 3). Other sites also contribute to this record of humanoccupation; a second locality at Happisburgh, referred to as Site 1, attests to human presence at around500 ka (MIS 13). This paper provides the first comprehensive account of research undertaken at HappisburghSite 1 since 2000. The early human landscape and depositional environment was that of a riverfloodplain, where an active river channel, in which a grey sand was deposited, was abandoned, forming afloodplain lake, with marginal marsh/swamp environments, which was infilled with organic mud. Thissuccession is sealed by Middle Pleistocene glacial deposits. An assemblage of 199 flint flakes, flake toolsand cores was recovered from the grey sand and organic mud. The evidence from Happisburgh Site 1 isplaced in the context of the wider British and European MIS 13 record. The growing evidence for asignificant dispersal of humans into northern Europe around 500 ka raises critical questions concerningthe environmental conditions under which this took place. We also consider the evolutionary andbehavioural changes in human populations that might have enabled the more widespread and persistentperiod of human presence in northern Europe at this time.</p

Identifying and prioritising unanswered research questions for people with hyperacusis: James Lind Alliance Hyperacusis Priority Setting Partnership

Objective To determine research priorities in hyperacusis that key stakeholders agree are the most important. Design/setting A priority setting partnership using two international surveys, and a UK prioritisation workshop, adhering to the six-staged methodology outlined by the James Lind Alliance. Participants People with lived experience of hyperacusis, parents/carers, family and friends, educational professionals and healthcare professionals who support and/or treat adults and children who experience hyperacusis, including but not limited to surgeons, audiologists, psychologists and hearing therapists. Methods The priority setting partnership was conducted from August 2017 to July 2018. An international identification survey asked respondents to submit any questions/uncertainties about hyperacusis. Uncertainties were categorised, refined and rephrased into representative indicative questions using thematic analysis techniques. These questions were verified as ‘unanswered’ through searches of current evidence. A second international survey asked respondents to vote for their top 10 priority questions. A shortlist of questions that represented votes from all stakeholder groups was prioritised into a top 10 at the final prioritisation workshop (UK). Results In the identification survey, 312 respondents submitted 2730 uncertainties. Of those uncertainties, 593 were removed as out of scope, and the remaining were refined into 85 indicative questions. None of the indicative questions had already been answered in research. The second survey collected votes from 327 respondents, which resulted in a shortlist of 28 representative questions for the final workshop. Consensus was reached on the top 10 priorities for future research, including identifying causes and underlying mechanisms, effective management and training for healthcare professionals. Conclusions These priorities were identified and shaped by people with lived experience, parents/carers and healthcare professionals, and as such are an essential resource for directing future research in hyperacusis. Researchers and funders should focus on addressing these priorities.Additional co-authors: Tracey Pollard, Helen Henshaw, Toto A Gronlund, Derek J Hoar

The clinical effectiveness and cost effectiveness of clozapine for inpatients with severe borderline personality disorder (CALMED study): A randomised placebo-controlled trial

Background: Data from case series suggest that clozapine may benefit inpatients with borderline personality disorder (BPD), but randomised trials have not been conducted. Methods: Multicentre, double-blind, placebo-controlled trial. We aimed to recruit 222 inpatients with severe BPD aged 18 or over, who had failed to respond to other antipsychotic medications. We randomly allocated participants on a 1:1 ratio to receive up to 400mg of clozapine per day or an inert placebo using a remote web-based randomisation service. The primary outcome was total score on the Zanarini Rating scale for Borderline Personality Disorder (ZAN-BPD) at six months. Secondary outcomes included self-harm, aggression, resource use and costs, side effects and adverse events. We used a modified intention to treat analysis (mITT) restricted to those who took one or more dose of trial medication, using a general linear model fitted at six months adjusted for baseline score, allocation group and site. Results: The study closed early due to poor recruitment and the impact of the COVID-19 pandemic. Of 29 study participants, 24 (83%) were followed up at six months, of whom 21 (72%) were included in the mITT analysis. At six months, 11 (73%) participants assigned to clozapine and 6 (43%) of those assigned to placebo were still taking trial medication. Adjusted difference in mean total ZAN-BPD score at six months was -3.86 (95% Confidence Intervals = -10.04 to 2.32, p=0.22). There were 14 serious adverse events; six in the clozapine arm and eight in the placebo arm of the trial. There was little difference in the cost of care between groups. Interpretation: We recruited insufficient participants to test the primary hypothesis. The study findings highlight problems in conducting placebo-controlled trials of clozapine and in using clozapine for people with BPD, outside specialist inpatient mental health units. Trial registration ISRCTN18352058. https://doi.org/10.1186/ISRCTN1835205

Superiority and cost-effectiveness of monthly extended-release buprenorphine versus daily standard of care medication: a pragmatic, parallel-group, open-label, multicentre, randomised, controlled, phase 3 trial

BACKGROUND: Daily methadone maintenance or buprenorphine treatment is the standard-of-care (SoC) medication for opioid use disorder (OUD). Subcutaneously injected, extended-release buprenorphine (BUP-XR) may be more effective-but there has been no superiority evaluation.METHODS: This pragmatic, parallel-group, open-label, multi-centre, effectiveness superiority randomised, controlled, phase 3 trial was conducted at five National Health Service community-based treatment clinics in England and Scotland. Participants (adults aged ≥ 18 years; all meeting DSM-5 diagnostic criteria for moderate or severe OUD at admission to their current maintenance treatment episode) were randomly assigned (1:1) to receive continued daily SoC (liquid methadone (usual dose range: 60-120 mg) or sublingual/transmucosal buprenorphine (usual dose range: 8-24 mg) for 24 weeks; or monthly BUP-XR (Sublocade;® two injections of 300 mg, then four maintenance injections of 100 mg or 300 mg, with maintenance dose selected by response and preference) for 24 weeks. In the intent-to-treat population (senior statistician blinded to blinded to treatment group allocation), and with a seven-day grace period after randomisation, the primary endpoint was the count of days abstinent from non-medical opioids between days 8-168 (i.e., weeks 2-24; range: 0-161 days). Safety was reported for the intention-to- treat population. Adopting a broad societal perspective inclusive of criminal justice, NHS and personal social service costs, a trial-based cost-utility analysis estimated the Incremental Cost-effectiveness Ratio (ICER) per quality-adjusted life year (QALY) of BUP-XR versus SoC at the National Institute for Health and Care Excellence threshold. The study was registered EudraCT (2018-004460-63) and ClinicalTrials.gov (NCT05164549), and is completed.FINDINGS: Between Aug 9, 2019 and Nov 2, 2021, 314 participants were randomly allocated to receive SoC (n = 156) or BUP-XR (n = 158). Participants were abstinent from opioids for an adjusted mean of 104.37 days (standard error [SE] 9.89; range: 0-161 days) in the SoC group and an adjusted mean of 123.43 days (SE 4.76; range: 24-161 days) in the BUP-XR group (adjusted incident rate ratio [IRR] 1.18, 95% confidence interval [CI] 1.05-1.33; p-value 0.004). The incidence of any adverse event was higher in the BUP-XR group than the SoC group (128 [81.0%] of 158 participants versus 67 [42.9%] of 156 participants, respectively-most commonly rapidly-resolving (mild-moderate range) pain from drug administration in the BUP-XR group (121 [26.9%] of 450 adverse events). There were 11 serious adverse events (7.0%) in the 158 participants in the BUP-XR group, and 18 serious adverse events (11.5%) in the 156 participants in the SoC group-none judged to be related to study treatment. The BUP-XR treatment group had a mean incremental cost of £1033 (95% central range [CR] -1189 to 3225) and was associated with a mean incremental QALY of 0.02 (95% CR 0.00-0.05), and an ICER of £47,540 (0.37 probability of being cost-effective at the £30,000/QALY gained willingness-to-pay threshold). However, BUP-XR dominated the SoC among participants who were rated more severe at study baseline, and among participants in maintenance treatment for more that 28 days at study enrolment.INTERPRETATION: Evaluated against the daily oral SoC, monthly BUP-XR is clinically superior, delivering greater abstinence from opioids, and with a comparable safety profile. BUP-XR was not cost-effective in a base case cost-utility analysis using the societal perspective, but it was more effective and less costly (dominant) among participants with more severe OUD, or those whose current treatment episode was longer than 28 days. Further trials are needed to evaluate if BUP-XR is associated with better clinical and health economic outcomes over the longer term.FUNDING: Indivior.</p

Extended-release pharmacotherapy for opioid use disorder (EXPO):protocol for an open-label randomised controlled trial of the effectiveness and cost-effectiveness of injectable buprenorphine versus sublingual tablet buprenorphine and oral liquid methadone

BACKGROUND: Sublingual tablet buprenorphine (BUP-SL) and oral liquid methadone (MET) are the daily, standard-of-care (SOC) opioid agonist treatment medications for opioid use disorder (OUD). A sizable proportion of the OUD treatment population is not exposed to sufficient treatment to attain the desired clinical benefit. Two promising therapeutic technologies address this deficit: long-acting injectable buprenorphine and personalised psychosocial interventions (PSI). This study will determine (A) the effectiveness and cost-effectiveness — monthly injectable, extended-release (BUP-XR) in a head-to-head comparison with BUP-SL and MET, and (B) the effectiveness of BUP-XR with adjunctive PSI versus BUP-SL and MET with PSI. Safety, retention, craving, substance use, quality-adjusted life years, social functioning, and subjective recovery from OUD will be also evaluated. METHODS: This is a pragmatic, multi-centre, open-label, parallel-group, superiority RCT, with a qualitative (mixed-methods) evaluation. The study population is adults. The setting is five National Health Service community treatment centres in England and Scotland. At each centre, participants will be randomly allocated (1:1) to BUP-XR or SOC. At the London study co-ordinating centre, there will also be allocation of participants to BUP-XR with PSI or SOC with PSI. With 24 weeks of study treatment, the primary outcome is days of abstinence from non-medical opioids during study weeks 2–24 combined with up to 12 urine drug screen tests for opioids. For 90% power (alpha, 5%; 15% inflation for attrition), 304 participants are needed for the BUP-XR versus SOC comparison. With the same planning parameters, 300 participants are needed for the BUP-XR and PSI versus SOC and PSI comparison. Statistical and health economic analysis plans will be published before data-lock on the Open Science Framework. Findings will be reported in accordance with the Consolidated Standards of Reporting Trials and Consolidated Health Economic Evaluation Reporting Standards. DISCUSSION: This pragmatic randomised controlled trial is the first evaluation of injectable BUP-XR versus the SOC medications BUP-SL and MET, with personalised PSI. If there is evidence for the superiority of BUP-XR over SOC medication, study findings will have substantial implications for OUD clinical practice and treatment policy in the UK and elsewhere. TRIAL REGISTRATION: EU Clinical Trials register 2018-004460-63. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13063-022-06595-0