The use of gene expression profiling to identify novel minimal residual disease markers (MRD) in acute myeloid leukaemia (AML)

Acute myeloid leukaemia (AML) is a heterogeneous disorder characterised by the accumulation of immature haematopoietic cells blocked at various stages of differentiation. Despite improved survival rates over the past decade, relapse occurs in approximately 70% patients undergoing chemotherapy. A potential reason for this is that current clinical protocols do not take account of the level of residual disease present at remission. Therefore, one strategy to reduce relapse rates is to monitor minimal residual disease and continue to treat until the patient is minimal residual disease negative. Current minimal residual disease markers are available for patients with characterised fusion genes but approximately 50% of patients have no detectable chromosomal aberration and therefore are without markers. Gene expression profiling is a powerful tool for disease classification, prognosis and therapeutic predictions. This study aimed to investigate the use gene expression profiling to identify novel minimal residual disease markers for specific AML sub-groups. Patient diagnostic samples were profiled to identify genes specific to AML patients with a favourable translocation in order to establish the "proof-of-principle". Several genes identified were followed in patient diagnostic and follow- up samples and compared to the markers currently used. Continuing with normal karyotype AML, genes were identified as specific to this sub-group. Several homeobox (HOX) genes and the Wilms' tumour (WT1) gene were identified and their MRD levels followed in diagnostic and follow-up samples. Only WT1 identified as specific to normal karyotype AML met the necessary criteria to be an MRD marker. Although the majority of genes selected from the GEP in this study proved unsuitable as markers, the identification and validation of a marker already used for MRD monitoring, WT1, demonstrates the ability of gene expression profiling to identify potential minimal residual disease markers in normal karyotype AML

Rip currents in the UK: incident analysis, public awareness, and education

Chapter 3 has been published as conference proceedings in the Journal of Coastal Research, and Chapter 4 has been published in the International Journal of Aquatic Research and EducationRip currents present a severe hazard to water users worldwide, resulting in over 100 drownings and thousands of lifeguard rescues annually. This thesis examines the demographics of who is effected by rip currents in the UK, what activity they are undertaking, when and where incidents occur, how much the public know, what people have experienced, and how best to educate them. Analysis of 7909 rip current lifeguard rescues (16777 people) across the UK between 2006-2013 highlighted the most at risk group, and subsequent target audience for education, to be male teenagers aged between 13-17 years old (n=2906, 17%). Geographically, the highest incidents occurred on the beaches in the Southwest of England (n=6911, 87%). Incidents mostly occurred outside of lifeguard flagged areas (n=4302, 54%) and mainly involved those using bodyboards (n=5290, 52%). Through the analysis of 407 public beach-based rip current and beach safety questionnaires, it was established that beach users have a poor understanding of rip currents (n=263, 65%) but a good perception of the beach safety flags (n=389, 96%). People with greater knowledge were typically educated by a lifeguard, enter the sea more frequently or have been caught in rip currents themselves. The experiences of 553 people caught in rip currents were analysed using an online questionnaire. The gender split was 69% male (n= 382) to 31% female (n= 171), indicating that males are caught in rip currents more than females. Swimming directly to shore against the rip followed an initial panic (n=108, 34%) for most people caught in a rip. Swimming parallel to the beach was the most remembered, advised, utilised, and promoted safety message. Respondents advocated the use of lifeguards to disseminate rip current safety messages. A new and unique rip current education programme was developed from the synthesis of these results. A lifeguard delivered a pilot programme to 185 teenagers in three schools and two community groups in the Southwest of England. This interactive pilot consisted of exercises using videos, photographs, news reports, and a swimming machine. Levels of rip current knowledge were evaluated before and after, and at regular intervals, to assess knowledge retention. The short-term effects after 3 months were positive, showing statistically significant (p<0.0005) improvements in mean knowledge levels. This thesis provides a new contribution to the expanding field of social and behavioural rip current research. The development of a unique rip current education programme presents an alternative method for increasing public awareness, and supports the worldwide prevention of rip current incidents and fatalities.Royal National Lifeboat Institution and The Marine Institut

Public Understanding and Knowledge of Rip Currents and Beach Safety in the UK

Rip currents present a severe hazard for water users on beaches and account for the greatest cause of lifeguard rescues worldwide. The physical dynamics of rip currents are well studied, and more recently, the social and behavioural science research surrounding human interaction of rip currents has been expanding, providing a social perspective and feeding into public education strategies. The aim of this study was to assess levels of public understanding of rip currents and beach safety on UK beaches. A questionnaire was undertaken (N = 407) during the summer of 2012 on four beaches. Beach users had a poor knowledge of rip currents (n = 263), but those who have been caught in a rip have a higher level of knowledge. Conversely, beach users had a good understanding of what the beach safety flags indicated (n = 314), and most people complied with this flag system (n = 339). In addition, people previously educated on rip currents had a higher knowledge, and lifeguards proved to be the most effective form of education. The study presents an insight into UK beach users’ knowledge of rip currents and provides more evidence with which to pilot a rip current education scheme within the UK

Public Understanding and Knowledge of Rip Currents and Beach Safety

Rip currents present a severe hazard for water users on beaches and account for the greatest cause of lifeguard rescues worldwide. The physical dynamics of rip currents are well studied, and more recently, the social and behavioural science research surrounding human interaction of rip currents has been expanding, providing a social perspective and feeding into public education strategies. The aim of this study was to assess levels of public understanding of rip currents and beach safety on UK beaches. A questionnaire was undertaken (N = 407) during the summer of 2012 on four beaches. Beach users had a poor knowledge of rip currents (n = 263), but those who have been caught in a rip have a higher level of knowledge. Conversely, beach users had a good understanding of what the beach safety flags indicated (n = 314), and most people complied with this flag system (n = 339). In addition, people previously educated on rip currents had a higher knowledge, and lifeguards proved to be the most effective form of education. The study presents an insight into UK beach users’ knowledge of rip currents and provides more evidence with which to pilot a rip current education scheme within the UK

Technicalities: Exploring the Labrador sea with autonomous vehicles

The Labrador Sea is a fascinating and difficult environment in which to work. In the winter, wind speeds can gust upwards of 200 km/hr, while 10-m wave heights and below freezing temperatures (-20°C) are not unheard off, making it an inhospitable area for field work. Indeed, few ships are present in the Labrador Sea during the winter. However, the same harsh conditions have made the Labrador Sea a key region for Earth’s climate, with the wintertime conditions resulting in localized deep mixing of waters and carbon to great depths (2 km) in the ocean [Lazier, 1980; Pickart, 1997]. As a consequence, in-situ observations in the Labrador Sea are critical to advancing scientific knowledge on past and future climate change scenarios. Previous attempts to use ships for wintertime work required long expeditions at sea, but often with little data collected due to unworkable conditions. Autonomous marine vehicles provide an obvious solution to collecting in-situ data in the wintertime, as they can operate in extreme conditions yet still give us the flexibility to adapt our sampling during the mission [deYoung et al., 2018; Testor et al., 2019]

Supporting medical students to support peers : a qualitative interview study

Background Students may be the first to recognise and respond to psychological distress in other students. Peer support could overcome medical student reluctance to seek help despite their high rates of mental ill-health. Yet, despite the adoption of peer support programs, there is little evidence of impact on students. Peer support programs may assume that medical students accept and view peer support positively. We explored these assumptions by asking students about their experiences and views on peer support. Methods Qualitative semi-structured interviews exploring peer support experiences and views on peer support were conducted with ten medical students at two contrasting medical schools. Informed by a constructivist stance, interview transcripts underwent thematic analysis. Results Three groups of themes were identified: participants' experiences of peer support encounters, concerns about providing support, and views on students' roles in peer support. Participants readily recalled signs of peer distress. Encounters were ad hoc, informal, and occurred within relationships based on friendship or by being co-located in the same classes or placements. Concerns about initiating and offering support included lack of expertise, maintaining confidentiality, stigma from a mental health diagnosis, and unclear role boundaries, with implications for acceptance of student roles in peer support. Conclusions Our study emphasised the centrality of social relationships in enabling or discouraging peer support. Relationships developed during medical studies may anticipate the collegial relationships between medical professionals. Nevertheless, only some students are willing to undertake peer support roles. We suggest different strategies for promoting informal peer support that can be offered by any student, to those promoting formal support roles for selected students. Future research focusing on the impact for both the students who receive, and on the students who provide peer support is called for

Public Understanding and Knowledge of Rip Currents and Beach Safety in the UK

Rip currents present a severe hazard for water users on beaches and account for the greatest cause of lifeguard rescues worldwide. The physical dynamics of rip currents are well studied, and more recently, the social and behavioural science research surrounding human interaction of rip currents has been expanding, providing a social perspective and feeding into public education strategies. The aim of this study was to assess levels of public understanding of rip currents and beach safety on UK beaches. A questionnaire was undertaken (N = 407) during the summer of 2012 on four beaches. Beach users had a poor knowledge of rip currents (n = 263), but those who have been caught in a rip have a higher level of knowledge. Conversely, beach users had a good understanding of what the beach safety flags indicated (n = 314), and most people complied with this flag system (n = 339). In addition, people previously educated on rip currents had a higher knowledge, and lifeguards proved to be the most effective form of education. The study presents an insight into UK beach users’ knowledge of rip currents and provides more evidence with which to pilot a rip current education scheme within the UK

Protocol for Delivery of CRISPR/dCas9 Systems for Epigenetic Editing into Solid Tumors Using Lipid Nanoparticles Encapsulating RNA

Genome editing tools, particularly the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems (e.g., CRISPR/Cas9), and their repurposing into epigenetic editing platforms, offer enormous potential as safe and customizable therapies for cancer. Specifically, various transcriptional abnormalities in human malignancies, such as silencing of tumor suppressors and ectopic re-expression of oncogenes, have been successfully targeted with virtually no off-target effects using CRISPR activation and repression systems. In these systems, the nuclease-deactivated Cas9 protein (dCas9) is fused to one or more domains inducing selective activation or repression of the targeted genes. Despite these advances, the efficient in vivo delivery of these molecules into the target cancer cells represents a critical barrier to accomplishing translation into a clinical therapy setting for cancer. Major obstacles include the large size of dCas9 fusion proteins, the necessity of multimodal delivery of protein and gRNAs, and the potential of these formulations to elicit detrimental immune responses.In this context, viral methods for delivering CRISPR face several limitations, such as the packaging capacity of the viral genome, the potential for integration of the nucleic acids into the host cells genome, and immunogenicity of viral proteins, posing serious safety concerns. The rapid development of mRNA vaccines in response to the COVID-19 pandemic has rekindled interest in mRNA-based approaches for CRISPR/dCas9 delivery. Simultaneously, due to their high loading capacity, scalability, customizable surface modification for cell targeting, and low immunogenicity, lipid nanoparticles (LNPs) have been widely explored as nonviral vectors. In this chapter, we first describe the design of optimized dCas9-effector mRNAs and gRNAs for epigenetic editing. We outline formulations of LNPs suitable for dCas9 mRNA delivery. Additionally, we provide a protocol for the co-encapsulation of the dCas9-effector mRNAs and gRNA into these LNPs, along with detailed methods for delivering these formulations to both cell lines (in vitro) and mouse models of breast cancer (in vivo)