A quality-by-design approach to improve process understanding and optimise the production and quality of CAR-T cells in automated stirred-tank bioreactors.

Ex vivo genetically-modified cellular immunotherapies, such as chimeric antigen receptor T cell (CAR-T) therapies, have generated significant clinical and commercial outcomes due to their unparalleled response rates against relapsed and refractory blood cancers. However, the development and scalable manufacture of these novel therapies remains challenging and further process understanding and optimisation is required to improve product quality and yield. In this study, we employ a quality-by-design (QbD) approach to systematically investigate the impact of critical process parameters (CPPs) during the expansion step on the critical quality attributes (CQAs) of CAR-T cells. Utilising the design of experiments (DOE) methodology, we investigated the impact of multiple CPPs, such as number of activations, culture seeding density, seed train time, and IL-2 concentration, on CAR-T CQAs including, cell yield, viability, metabolism, immunophenotype, T cell differentiation, exhaustion and CAR expression. Initial studies undertaken in G-Rex® 24 multi-well plates demonstrated that the combination of a single activation step and a shorter, 3-day, seed train resulted in significant CAR-T yield and quality improvements, specifically a 3-fold increase in cell yield, a 30% reduction in exhaustion marker expression and more efficient metabolism when compared to a process involving 2 activation steps and a 7-day seed train. Similar findings were observed when the CPPs identified in the G-Rex® multi-well plates studies were translated to a larger-scale automated, controlled stirred-tank bioreactor (Ambr® 250 High Throughput) process. The single activation step and reduced seed train time resulted in a similar, significant improvement in CAR-T CQAs including cell yield, quality and metabolism in the Ambr® 250 High Throughput bioreactor, thereby validating the findings of the small-scale studies and resulting in significant process understanding and improvements. This study provides a methodology for the systematic investigation of CAR-T CPPs and the findings demonstrate the scope and impact of enhanced process understanding for improved CAR-T production.

Island communities and disaster resilience: Applying the EnRiCH community resilience framework.

OBJECTIVE: To explore the beliefs, attitudes, and perspectives of community resilience in St. Kitts and Nevis. DESIGN: Qualitative Interpretive Phenomenological Analysis using the EnRiCH Community Resilience Framework for High-Risk Populations (EnRiCH Framework) to identify factors that enhance or create barriers to community resilience to disasters in St. Kitts and Nevis. SAMPLE: Twenty-one key informants and 23 community informants provided insight into the history of disasters. RESULTS: Unique strengths and barriers that significantly influenced this high-risk population's adaptive capacity were identified. A discrepancy between the way disaster preparedness was perceived by government officials and the local population was noted. Cultural factors promoted connectedness and communication and created barriers to empowerment and collaboration. Innovative strategies were suggested that could enhance upstream leadership, downstream management, and resource management during disasters. CONCLUSIONS: Island communities represent a unique opportunity to examine risk reduction and vulnerability within the context of community and societal characteristics. This research addresses a significant gap in the literature on interventions that utilize a strengths-based approach to building adaptive capacity and resilience to disasters among at-risk populations. The EnRiCH Framework can be used to develop an approach to strengthen adaptive capacity and improve resilience to disasters.

Student Nurses' Experiences of Critical Events in the Clinical Setting: A Grounded Theory.

BACKGROUND: Nursing students often experience critical events in the clinical setting and clinical instructors may not be prepared to adequately support them. These students often feel alone and abandoned, increasing their risk of psychological distress. PURPOSE: A grounded theory study was conducted to explore pre-licensure nursing students' experiences of critical events in the clinical setting. Specific attention was paid to understanding how they are prepared for and supported before, during, and after critical events, and their experiences of psychological distress and psychological recovery. METHOD: Strauss and Corbin's Grounded Theory methodology was used for this study. RESULTS: Fourteen undergraduate student nurses from universities in the western United States were interviewed. Ten concepts were identified, and a theory of Student Nurses' Experiences of Critical Events in the Clinical Setting emerged. CONCLUSION: Findings indicate that student nurses need active instructor and/or staff support during critical events, and pre-briefing whenever possible. Debriefing positively affected students' post-event stress response and lack of debrief was associated with post-event psychological distress. Support for students exposed to critical events during clinical experiences should continue in the days, weeks, and months following the event; clinical instructors need to be prepared to provide this support. All faculty members should monitor students for signs of psychological distress and better support students' mental health and emotional wellbeing.

Thermally drawn fibers as nerve guidance scaffolds.

Synthetic neural scaffolds hold promise to eventually replace nerve autografts for tissue repair following peripheral nerve injury. Despite substantial evidence for the influence of scaffold geometry and dimensions on the rate of axonal growth, systematic evaluation of these parameters remains a challenge due to limitations in materials processing. We have employed fiber drawing to engineer a wide spectrum of polymer-based neural scaffolds with varied geometries and core sizes. Using isolated whole dorsal root ganglia as an in vitro model system we have identified key features enhancing nerve growth within these fiber scaffolds. Our approach enabled straightforward integration of microscopic topography at the scale of nerve fascicles within the scaffold cores, which led to accelerated Schwann cell migration, as well as neurite growth and alignment. Our findings indicate that fiber drawing provides a scalable and versatile strategy for producing nerve guidance channels capable of controlling direction and accelerating the rate of axonal growth.

Prefilled devices for parenteral applications.

Current parenteral administration of drugs suffers from several drawbacks including the requirement of healthcare personnel to administer the drug, the risk of needle stick injuries that may result in the transmission of blood borne pathogens, and patient discomfort. Prefilled devices have emerged as powerful tools to improve parenteral administration of drugs. There are a number of clinical conditions including treatment of endocrine diseases, neurological disorders, autoimmune diseases and emergency medicine where prefilled devices have made major improvements to patient care. Prefilled devices have become an important set of tools for the medical practitioner due to their ease of use and safety, cost effectiveness and patient convenience. This review provides a comprehensive summary of existing prefilled devices, their current clinical uses and corresponding regulatory processes.