Simulation and characterization of the corneal limbal epithelial stem cell niche by using new biotechnologies

The limbal epithelial stem cell (LESC) niche is a vital stem cell pool which serves to replenish the cornea with competent stem cells for regrowth and repair throughout adult life. Injury and disease, such as limbal stem cell deficiency (LSCD), can damage and deplete this stem cell pool, resulting in aberrant growth across the cornea and can lead to blindness. The aims of this thesis are the investigation and replication of the LESC niche. Investigation of the niche provided valuable insight into the combined structural and mechanical properties of the niche. Meanwhile, the replication of the niche using smart material fabrication methods enabled active manipulation of primary isolated limbal cells to model niche conditions. These aims were achieved by focusing on the mechanical characterisation of human limbal tissues by non-destructive optical coherence elastography (OCE), combined with optical coherence tomography (OCT) and anatomic replication through the development of a novel bioreactor system. The first study, using OCT/OCE characterisation, revealed the limbal undulation architecture of palisades of Vogt with alteration (in dimension and modulus) in key anatomical features with ageing. To produce a biomimetic in vitro model of the limbus, polydimethylsiloxane (PDMS) based substrates with wrinkled topography were generated by plasma treatment, acid oxidation and dual treatments using a novel stretching frame. This system enabled the PDMS substrate to flatten and wrinkle in a reversible pattern when conducting cell culture, forming the culture surface of a new type of bioreactor. The crypt-like pattern’s dimensions resembled the topography of the LESC niche. The biocompatibility of the patterned substrate was markedly improved using a Gelatin Methacrylate (GelMa) gel coating. The limbal cells cultured on the wrinkled topography proved to retain stemness through the preservation of key stem cell markers such as ABCG2 and P63, whilst indicating the induction of epithelial change by increases in CK3 expression. It was also observed that these wrinkled PDMS surfaces were able to dictate cell growth patterns, showing alignment in motile cells and colony segregation in colony-forming cells in human and porcine limbal cells respectively. The biotechnology developed in this research has exciting potential applications as a disease model for conditions such as LSCD, ageing or injuries where substantial physical anatomical changes can be investigated for their effect on the native cell population. In translational terms, this benchtop application has the potential to reduce the dependency on patient study cases to investigate limbal and other optical surface diseases by providing an actively tuneable culture platform which has a small footprint. Additionally, this technology has scope to reduce the dependency on primary tissue isolations by providing a culture platform whose topographical features can be tailor-made to expansion conditions

Feasibility and Acceptability of Online Culturally Adapted Cognitive Behavioural Therapy for Depression and Anxiety in Canadians of South Asian Origin: A Randomized Controlled Trial: Faisabilité et acceptabilité de la thérapie cognitivo-comportementale en ligne adaptée à la culture pour traiter la dépression et l'anxiété chez les Canadiens d'origine sud-asiatique : Essai contrôlé à répartition aléatoire.

BackgroundThis paper reports a pilot trial of culturally adapted CBT (CaCBT) for Canadian South Asians. The primary objective of this study was to assess the feasibility and acceptability of online CaCBT to treat anxiety and depression in Canadian South Asian individuals. The secondary objective was to measure changes in depression, anxiety, and disability.MethodsAn assessor-blind randomized clinical trial was conducted at 3 sites in Canada (Greater Toronto Area, Ottawa, and Vancouver). One hundred forty-six participants were randomly allocated to 1 of 2 groups: Ca-CBT (experimental group) or standard cognitive behavioural therapy (CBT) (control group) in a 1:1 ratio. The primary outcome, feasibility, was measured through engagement, recruitment, and participant retention. Acceptability was measured through the Verona Service Satisfaction Scale. Working Alliance Inventory was used to measure therapeutic engagement. Secondary outcomes included depression (Hospital Anxiety and Depression Scale-HADS), somatic symptoms (Bradford Somatic Inventory-BSI), and disability (WHO Disability Assessment Schedule 2.0 (WHODAS). Assessments were carried out at baseline, at the end of therapy (12 weeks from baseline), and at follow-up (36 weeks from baseline).ResultsWe were able to recruit participants within the given timeframe with excellent retention rates in both arms. Most participants in the intervention group, 56 (74.7%), attended ≥ 8 sessions, and 11 (14.7%) attended 5 to 7 sessions. Eight (10.7%) participants from the intervention group and 9 (12.0%) from the control group dropped out of therapy (<5 sessions). Participants in the intervention group reported higher levels of satisfaction (  = 0.001) and therapeutic engagement (  < 0.001) compared with the control group. Participants in both groups benefited from CBT.ConclusionsThis is the first report of online CaCBT for depression and anxiety for Canadian South Asians. The intervention is acceptable and feasible. Culturally adapted CBT is as effective as standard CBT in reducing the symptoms of depression and anxiety

Germline transformation of the West Nile virus and avian malaria vector Culex quinquefasciatus Say using the piggyBac transposon system.

Culex quinquefasciatus Say is a mosquito which acts as a vector for numerous diseases including West Nile virus, lymphatic filariasis and avian malaria, over a broad geographical range. As the effectiveness of insecticidal mosquito control methods declines, the need has grown to develop genetic control methods to curb the spread of disease. The piggyBac transposon system - the most widely used genetic transformation tool in insects, including mosquitoes - generates quasi-random insertions of donor DNA into the host genome. However, despite the broad reported species range of piggyBac, previous attempts to use this tool to transform Culex quinquefasciatus mosquitoes have failed. Here we report the first successful transformation of Culex quinquefasciatus with the piggyBac transposon system. Using commercially synthesised piggyBac mRNA as a transposase source, we were able to generate three independent insertions of a ZsGreen fluorescent marker gene, with transformation efficiencies of up to 5 %. Through this work, we have expanded the genetic toolkit available for the genetic manipulation of Culex mosquitoes and thus removed a barrier to developing novel genetic control methods in this important disease vector. [Abstract copyright: Copyright © 2025. Published by Elsevier Ltd.

A scoping review of evidence for the effects of seven global deer species on woody vegetation

Context: Rapid expansion of deer (Cervidae) populations is a concern for forest ecosystems. Despite extensive reviews on how deer affect forests, variation in effects across deer species has received less attention. A lack of focus on species‐specific effects may lead to oversights and failure to achieve desired management outcomes. Methodology: We used a systematic approach to compile data on the extent to which the effects of seven deer species on woody vegetation have been studied. We focused on the six deer species present in Britain and Ireland, and elk (Cervus canadensis). Results: A total of 455 studies were included from across the globe. Red deer (Cervus elaphus) (n = 163) and elk (n = 158) were the most studied species, while Reeve's muntjac (Muntiacus reevesi) (n = 18) and Chinese water deer (Hydropotes inermis) (n = 5) were the least researched. Fifty‐four per cent of studies (n = 245) used fenced exclosures to assess deer impacts. Research mainly focused on defoliation via browsing and grazing (n = 424), while debarking (n = 44), defecation (n = 8) and trampling (n = 5) were less frequently studied. Vegetation density (n = 235), height (n = 189) and diversity (n = 135) were the most common metrics used, while fewer studies focused on vegetation mortality (n = 74), structural variability (n = 28) and condition (n = 15). Practical implication: While previous studies have often focused on the probability or severity of deer damage to woody vegetation, we identified key knowledge gaps on the ecological influence of such damage, with a species‐specific focus. Researchers should treat deer species as distinct entities and appreciate the differences in their body size, sociality, physiology and behaviour when studying their ecological effects. Where multiple deer species co‐occur, identifying relative local species abundance and differences among species foraging behaviours will help to determine how their interactions—whether additive, synergistic or antagonistic—affect ecosystem processes and vegetation dynamics

A humanised thrombus-on-a-chip model utilising tissue-engineered arterial constructs: A method to reduce and replace mice used in thrombosis and haemostasis research.

The study of in vivo thrombus formation has principally been performed using intravital microscopy in mice and other species. These have allowed us to visualise the molecular and cellular processes that regulate thrombus formation inside the body. However current in vivo arterial thrombosis models are difficult to standardise between labs and frequently produce results that do not reliably translate successfully in human clinical trials. Here we provide a step-by-step description with accompanying video tutorials to demonstrate how to produce a 3D humanised thrombus-on-a-chip model, which uses perfusion of fluorescently-labelled human blood over a mechanically-injured human tissue engineered arterial construct (TEAC) within a 3D printed microfluidic flow chamber to replicate thrombus formation within a healthy artery. We also provide a written methodology on how to use 3D printing to produce a mechanical injury press that can reproducibly damage the TEAC as a stimulus for thrombus formation as part of a mechanical injury model. Perfusion of the uninjured TEAC with whole human blood containing DiOC6-labelled platelets without initiating notable thrombus formation. The mechanical injury press was shown to induce a reproducible puncture wound in the TEAC. Fluorescence microscopy was used to demonstrate that thrombus formation could be observed reproducibly around sites of injury. This humanised thrombosis-on-a-chip model can replace the use of animals in in vivo thrombosis models for preclinical assessment of anti-thrombotic therapies. This method also offers multiple scientific advantages: allowing new drugs to be directly tested on human blood from a diverse array of donors, facilitating use of a realistic and reproducible injury modality as well as removing the potential confounding effects of general anaesthetics in animal studies. The use of human thrombus-on-a-chip models combining TEACs offers a new methodology to reduce animal use whilst improving the predictive capabilities of preclinical trials of anti-thrombotic therapies

Understanding internet-supported self-management for low back pain in primary care: a qualitative process evaluation of the SupportBack 2 randomised controlled trial

Objective: The SupportBack 2 randomised controlled trial (RCT) compared the clinical and cost-effectiveness of an internet intervention supporting self-management versus usual primary care in reducing low back pain (LBP)-related disability. In this study, we aimed to identify and understand key processes and potential mechanisms underlying the impact of the intervention. Design: This was a nested qualitative process evaluation of the SupportBack 2 RCT (ISRCTN: 14736486 pre-results). Setting: Primary care in the UK (England). Participants: 46 trial participants experiencing LBP without indicators of serious spinal pathologies (eg, fractures, infection) took part in telephone interviews at either 3 (n=15), 6 (n=14) or 12 months (n=17) post randomisation. Five physiotherapists who provided telephone support for the internet intervention also took part in telephone interviews. Intervention: An internet intervention ‘SupportBack’ supporting self-management of LBP primarily through physical activity and exercise delivered in addition to usual care, with and without physiotherapist telephone support. Analysis: Data were analysed thematically, applying a realist logic to develop context-mechanism-outcome configurations. Results: Four explanatory themes were developed, with five context-mechanism-outcome configurations. Where benefit was reported, SupportBack appeared to work by facilitating a central associative process where participants linked increases in physical activity or exercise with improvements in LBP, then continued to use physical activity or exercise as key regulatory strategies. Participants who reported little or no benefit from the intervention appeared to experience several barriers to this associative process, including negative expectations, prohibitive beliefs about the cause of LBP or functional limitations preventing engagement. Physiotherapists appeared to provide accountability and validation for some; however, the remote telephone support that lacked physical assessment was viewed as limiting its potential value. Conclusions: Digital interventions targeting physical activity and exercise to support LBP self-management may rely on mechanisms that are easily inhibited in complex, heterogeneous populations. Future research should focus on identifying and removing barriers that may limit the effectiveness of digital self-management support for LBP

Modular Monolith Architecture in Cloud Environments: A Systematic Literature Review

Modular monolithic architecture (MMA) has recently emerged as a hybrid architecture that is positioned between traditional monoliths and microservices. It combines operational simplicity with modularity and maintainability. Although industry adoption of the architecture is growing, academic research on MMA remains fragmented and lacks systematic synthesis. This paper presents the first systematic literature review (SLR) of MMA in cloud environments. The review follows Kitchenham’s guidelines; we searched six major digital libraries for peer-reviewed studies published between 2020 and May 2025. From 369 retrieved records, we included 15 primary studies through a structured review protocol. Our synthesis highlights the problem of inconsistent terminology usage in the literature. It also identifies the architectural scope of MMA, and specifies the adoption drivers such as simplified deployment, maintainability, and reduced orchestration overhead. We also analyse implementation practices—including Domain-Driven Design (DDD), modular boundaries, and containerised deployment—and highlight comparative evidence showing MMA’s suitability when microservices introduce excessive complexity or costs. Key research gaps include the absence of consensus on a clear comprehensive definition, limited empirical benchmarking, and insufficient tools support. Thus, this study establishes a conceptual foundation for future research and provides practitioners with structured insights to inform architectural decisions in cloud-native environments

Fear conditioning: Insights into learning, memory and extinction and its relevance to clinical disorders

Fear, whether innate or learned, is an essential emotion required for survival. The learning, and subsequent memory, of fearful events enhances our ability to recognise and respond to threats, aiding adaptation to new, ever-changing environments. Considerable research has leveraged associative learning protocols such as contextual or auditory forms of fear conditioning in rodents, to understand fear learning, memory consolidation and extinction phases of memory. Such assays have led to detailed characterisation of the underlying neurocircuitry and neurobiology supporting fear learning processes. Given fear processing is conserved across rodents and humans, fear conditioning experiments provide translational insights into fundamental memory processes and fear-related pathologies. This review examines associative learning protocols used to measure fear learning, memory and extinction, before providing an overview on the underlying complex neurocircuitry including the amygdala, hippocampus and medial prefrontal cortex. This is followed by an in-depth commentary on the neurobiology, particularly synaptic plasticity mechanisms, which regulate fear learning, memory and extinction. Next, we consider how fear conditioning assays in rodents can inform our understanding of disrupted fear memory in human disorders such as post-traumatic stress disorder (PTSD), anxiety and psychiatric disorders including schizophrenia. Lastly, we critically evaluate fear conditioning protocols, highlighting some of the experimental and theoretical limitations and the considerations required when conducting such assays, alongside recent methodological advancements in the field. Overall, rodent-based fear conditioning assays remain central to making progress in uncovering fundamental memory phenomena and understanding the aetiological mechanisms that underpin fear associated disorders, alongside the development of effective therapeutic strategies

3′-O-β-Glucosylation of nucleoside analogues using a promiscuous bacterial glycosyltransferase

Nucleoside analogue therapeutics have a proven capability within drug discovery as antiviral and antineoplastic agents. However, their efficacy can be limited by poor cellular uptake, off target toxicity and low bioavailability. Glycosylation of pharmaceutical agents/natural products represents a strategically simple method to modulate pharmacological profiles. Herein, we explore biocatalytic glycosylation of nucleoside analogues. The activity of the nucleoside-specific 3'-O-glycosyltransferase AvpGT from Streptomyces sp. AVP053U2 is investigated toward a panel of both natural and clinically relevant purine and pyrimidine nucleoside analogues. AvpGT demonstrates broad substrate promiscuity, with glycosylation observed by HILIC-MS for 15 of 21 nucleosides tested. Of these, 12 nucleosides were successfully glycosylated on ≥25 μmol scale in 39-91% isolated yields, including four current therapeutics