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Research has shown that English language proficiency (ELP) plays a crucial role in the psychological well-being of international students (e.g. Li et al., "Journal of International Students, "4(4), 301-313, 2014). However, few studies have reported positive correlations between their improved English skills and well-being. The present study examines how a new pedagogy called personalised autonomous (PA) model can induce a virtuous cycle in international PhD students to enhance their English language proficiency and consequently, their well-being. We collected both quantitative and qualitative data from two groups of students: those who experienced the PA model in a course entitled Personalised English Language Enhancement (PELE) and those who did not. We conducted surveys at the beginning and end of the term to compare entry and exit data. Comparison of the entry and exit survey data for PELE students showed significant improvements across almost all dependent variables including self-efficacy, well-being and a sense of belonging to the university community. In contrast, the non-PELE students did not significantly improve in any variable. In focus groups, students indicated how their improved communicative confidence sparked a virtuous cycle leading to greater well-being and a stronger sense of community. Our data strongly suggests that ELP extends beyond mere linguistic ability; it embodies a dynamic interplay between one's language skills and their self-confidence. Diminished confidence may trigger a vicious cycle of fear of communication, unwillingness to engage, unproductive research, and estranged relationships with supervisors. Therefore, it is imperative for educators and policymakers to critically review and enhance their approaches to supporting the ELP of international PhD students.

The pathophysiologic mechanism of sickle cell disease (SCD) involves polymerization of deoxygenated haemoglobin S (HbS), leading to red blood cell (RBC) sickling, decreased RBC deformability, microvascular obstruction, haemolysis, anaemia and downstream clinical complications. Pharmacological increase in the concentration of oxygenated HbS in RBCs has been shown to be a novel approach to inhibit HbS polymerization and reduce RBC sickling and haemolysis. We report that GBT021601, a small molecule that increases HbS-oxygen affinity, inhibits HbS polymerization and prevents RBC sickling in blood from patients with SCD. Moreover, in a murine model of SCD (SS mice), GBT021601 reduces RBC sickling, improves RBC deformability, prolongs RBC half-life and restores haemoglobin levels to the normal range, while improving oxygen delivery and increasing tolerance to severe hypoxia. Notably, oral dosing of GBT021601 in animals results in higher levels of Hb occupancy than voxelotor and suggests the feasibility of once-daily dosing in humans. In summary, GBT021601 improves RBC health and normalizes haemoglobin in SS mice, suggesting that it may be useful for the treatment of SCD. These data are being used as a foundation for clinical research and development of GBT021601.

Abstract Background Existing traditional cognitive screening tools for dementia have various limitations, including overreliance on tests assessing verbal memory and, to a lesser extent, on some aspects of executive functioning. Comprehensive neuropsychological assessment is sensitive to impairment but time-intensive and expensive. Virtual reality may provide a dynamic and unique understanding of cognitive performance and increase the ecological validity of cognitive assessment. The use of virtual reality in screening for cognitive function in older persons is promising, but evidence for its use remains sparse. Objective Our primary aim was to examine the feasibility and acceptability of a newly developed, virtual reality assessment module, ‘Leaf Café’, a computer-based program that assesses cognition in an engaging, efficient, and ecologically relevant way. The secondary aim was to assess the ability of the module to discriminate between performances of younger and older adults. Methods A cross-sectional study was carried out in Sydney, Australia, targeting adults aged 18 years and above. Participants completed a traditional cognitive screening tool (Telephone Interview for Cognitive Status-Modified, TICS-M) and Leaf Café, a low-immersive virtual reality module designed to evaluate learning and memory, perceptual-motor function, and executive functioning. The total performance score for each participant, ranging from 0 to 180, was correlated with their cognitive performance assessed by TICS-M, using Pearson’s correlation coefficient. Following module completion, participants were presented with an open and closed-question survey to capture their perceptions, attitudes, and feedback on the module, encompassing practicality, acceptability, and enjoyment. Both descriptive and content analyses were employed to interpret the obtained data. Results A sample of 131 participants (mean age 54.9 years, SD = 20.8, range 20–85) took part. The majority were female (71.8%) and born in an English-speaking country (75.8%). The mean amount of time spent in the module was 32.8 min (SD = 13.3) with a mean module score of 107.6 (SD = 38.7). Most participants completed the highest level (5; 80.5%). There was a significant correlation between Leaf Café total scores with TICS-M cognitive scores overall, and for both younger (aged 18–64 years) and older adult (aged 65 + years) groups. No significant difference was found on performance between age groups on TICS-M performance, however, younger adults had significantly better performance on the Leaf Café module than older adults (M = 124.1 vs 95.9; p

Abstract Background An informed understanding of older adults’ perceptions of the impact (positive or negative) of recurrent COVID-19 long lockdowns is important for the development of targeted interventions and resources for future restrictions. This study aimed to understand self-reported impacts of COVID-19 recurrent restrictions on older adults and how technology has been used to mitigate these. Methods A cross-sectional national study of 257 community-dwelling older Australians based in Victoria (mean age = 67.6 years [SD = 7.2]; 20.6% male) completed an online or postal survey as part of a larger study examining the physical and mental health impacts of a second extended COVID-19 lockdown period. This secondary analysis reports on the findings from free-text responses to two open-ended questions included in that survey that asked participants to comment on the greatest impacts of the COVID-19 lockdowns (positive or negative) and the role of technology in supporting their wellbeing during this time. Responses were collected between July and September 2020. Data were analysed using content (COVID-19 impacts) and thematic (role of technology) analysis. Results Respondents gave more negative responses (75.5%) than mixed (15.2%) and positive responses (6.2%) in reporting on the biggest impact of COVID-19 lockdowns. Inductive content analysis revealed two first-order main categories (Positive impacts and Negative impacts). Axial coding of main categories showed five second-order categories (Environmental, Physical Health, Social, Mental Health, and Personal) for both negative and positive main categories (totalling 10 second-order categories). Overall, respondents highlighted social loss as the key negative experience (70%), with acute feelings of social isolation contributing to negative impacts on mental wellbeing. The most commonly reported positive impact reported (11%) was having more time for relationships, relaxation, and new hobbies. Technology was primarily used to sustain socialisation and provide access to essential resources, services, and goods, which respondents perceived to contribute to maintaining their wellbeing. Conclusions Findings suggest a critical need for interventions that address the social loss experienced by older adults during COVID-19 recurrent lockdowns, particularly to alleviate the associated negative impact on mental wellbeing. Recognising the positive aspect of increased time for relationships and leisure activities indicates potential areas for resilience-building strategies. The pivotal role of technology in mitigating adverse effects highlights its significance in building social connections and supporting overall wellbeing during challenging times. These implications can guide future efforts to enhance older adults’ resilience, mental health, and holistic wellbeing in future public health crises.

The success of deep brain stimulation (DBS) for treating Parkinsons disease has led to its application to several other disorders, including treatment-resistant depression. Results with DBS for treatment-resistant depression have been heterogeneous, with inconsistencies largely driven by incomplete understanding of the brain networks regulating mood, especially on an individual basis. We report results from the first subject treated with DBS for treatment-resistant depression using an approach that incorporates intracranial recordings to personalize understanding of network behavior and its response to stimulation. These recordings enabled calculation of individually optimized DBS stimulation parameters using a novel inverse solution approach. In the ensuing double-blind, randomized phase incorporating these bespoke parameter sets, DBS led to remission of symptoms and dramatic improvement in quality of life. Results from this initial case demonstrate the feasibility of this personalized platform, which may be used to improve surgical neuromodulation for a vast array of neurologic and psychiatric disorders.

ABSTRACT Assuring that cell therapy products are safe before releasing them for use in patients is critical. Currently, compendial sterility testing for bacteria and fungi can take 7–14 days. The goal of this work was to develop a rapid untargeted approach for the sensitive detection of microbial contaminants at low abundance from low volume samples during the manufacturing process of cell therapies. We developed a long-read sequencing methodology using Oxford Nanopore Technologies MinION platform with 16S and 18S amplicon sequencing to detect USP organisms and other microbial species. Reads are classified metagenomically to predict the microbial species. We used an extreme gradient boosting machine learning algorithm (XGBoost) to first assess if a sample is contaminated, and second, determine whether the predicted contaminant is correctly classified or misclassified. The model was used to make a final decision on the sterility status of the input sample. An optimized experimental and bioinformatics pipeline starting from spiked species through to sequenced reads allowed for the detection of microbial samples at 10 colony-forming units (CFU)/mL using metagenomic classification. Machine learning can be coupled with long-read sequencing to detect and identify sample sterility status and microbial species present in T-cell cultures, including the USP organisms to 10 CFU/mL. IMPORTANCE This research presents a novel method for rapidly and accurately detecting microbial contaminants in cell therapy products, which is essential for ensuring patient safety. Traditional testing methods are time-consuming, taking 7–14 days, while our approach can significantly reduce this time. By combining advanced long-read nanopore sequencing techniques and machine learning, we can effectively identify the presence and types of microbial contaminants at low abundance levels. This breakthrough has the potential to improve the safety and efficiency of cell therapy manufacturing, leading to better patient outcomes and a more streamlined production process.

With the ever increasing pace of change in the Chemical and Pharmaceutical Industry, universities need to be more aware of the skill set that employers are seeking in graduates. These skills often include risk taking, creative thinking and entrepreneurship; skills that are not naturally associated with chemistry-related curriculums. This research focusses on the authors' experiences of applying for strategic grants which underpin the dissemination of such skills, and explores how these were used as the basis for creating specific learning content for final year students. Students' experiences after completing assignments were gathered through an online survey and subsequent analysis identified areas for improvement in chemistry curriculums.

Background: The current literature suggests a link between dummy (or pacifier) use and a number of both positive and detrimental consequences. Positive consequences include soothing effect and protection from sudden infant death syndrome (SIDS), while negative ones include increased risk of otitis media and dental malformation. However, there is little research surrounding the impact of dummy use on the development of speech sounds. Aims: To investigate whether duration (in number of months) and frequency per day of dummy use have an individual or combined effect on the development of a child's speech, and if so, in what way. Methods & Procedures: A total of 100 British-English children aged 24-61 months and growing up in the UK were recruited through nurseries, playgroups and by word of mouth. Their parents were asked to complete a questionnaire about the duration and frequency of dummy use and factors known to influence the development of speech. Following this, the children's speech was assessed using the phonology section of the Diagnostic Evaluation of Articulation and Phonology (DEAP). Analysis of the DEAP was conducted to determine the percentage of consonants correct, number of age-appropriate, delayed and atypical errors. Dummy use and speech outcome measures were then analysed qualitatively and quantitatively using mean and median group comparisons alongside multivariate generalized least squares and generalized negative binomial modelling approaches to test for significant associations. Outcomes & Results: The results showed that the majority of speech outcomes are not significantly associated with dummy use, however measured, in bivariate or multivariate analyses. However, there is a significant association between increased atypical errors and greater frequency of daytime dummy use. This association is strengthened by restricted sampling within the younger members of the sample, with this association not observable within children older than 38 months, the median sample age. Conclusions & Implications: The evidence base for any effects of dummy use on speech is very small. Dummy use may increase the number of atypical speech errors a young child makes. However, only the frequency of daytime use seems relevant, not the duration or night-time use, and these errors may resolve over time.

Double-stranded DNA (dsDNA) binding and cleavage by Cas9 is a hallmark of type II CRISPR-Cas bacterial adaptive immunity. All known Cas9 enzymes are thought to recognize DNA exclusively as a natural substrate, providing protection against DNA phage and plasmids. Here, we show that Cas9 enzymes from both subtypes II-A and II-C can recognize and cleave single-stranded RNA (ssRNA) by an RNA-guided mechanism that is independent of a protospacer-adjacent motif (PAM) sequence in the target RNA. RNA-guided RNA cleavage is programmable and site-specific, and we find that this activity can be exploited to reduce infection by single-stranded RNA phage in vivo. We also demonstrate that Cas9 can direct PAM-independent repression of gene expression in bacteria. These results indicate that a subset of Cas9 enzymes have the ability to act on both DNA and RNA target sequences, and suggest the potential for use in programmable RNA targeting applications.

Aim To quantify the areas of burden experienced by patients requiring repeated intravitreal injections (IVI) in the management of exudative retinal diseases.Methods The validated Questionnaire to Assess Life Impact of Treatment by Intravitreal Injections survey was administered to patients at four retina clinical practices across four US states. The primary outcome measure was Treatment Burden Score (TBS), a single score assessing overall burden.Results Of 1416 (n=657 age-related macular degeneration; n=360 diabetic macular oedema/diabetic retinopathy; n=221 retinal vein occlusion; n=178 other/uncertain) patients, 55% were women with an average age of 70 years. Patients most frequently reported receiving IVI every 4–5 weeks (40%). The mean TBS was 16.1±9.2 (range 1–48; scale of 1–54), and the TBS was higher in patients with diabetic macular oedema and/or diabetic retinopathy (DMO/DR) (17.1) compared with those with age-related macular degeneration (15.5) or retinal venous occlusive (15.3) (p=0.028). Though the mean level of discomfort was quite low (1.86) (scale 0–6), 50% of patients reported experiencing side effects more than half of the visits. Patients having received fewer than 5 IVI reported higher mean anxiety levels before (p=0.026), during (p=0.050) and after (p=0.016) treatment compared with patients having received more than 50 IVI. After the procedure, 42% of patients reported restrictions from usual activities due to discomfort. Patients reported a high mean satisfaction rating of 5.46 (scale 0–6) with the care of their diseases.Conclusions The mean TBS was moderate and highest among patients with DMO/DR. Patients with more total injections reported lower levels of discomfort and anxiety but higher disruption to daily life. Despite the challenges related to IVI, the overall satisfaction with treatment remained high.

Abstract The apelin receptor (Aplnr) and its ligands, Apelin and Apela, contribute to metabolic control. The insulin resistance associated with pregnancy is accommodated by an expansion of pancreatic β-cell mass (BCM) and increased insulin secretion, involving the proliferation of insulin-expressing, glucose transporter 2-low (Ins+Glut2LO) progenitor cells. We examined changes in the apelinergic system during normal mouse pregnancy and in pregnancies complicated by glucose intolerance with reduced BCM. Expression of Aplnr, Apelin and Apela was quantified in Ins+Glut2LO cells isolated from mouse pancreata and found to be significantly higher than in mature β-cells by DNA microarray and qPCR. Apelin was localized to most β-cells by immunohistochemistry although Aplnr was predominantly associated with Ins+Glut2LO cells. Aplnr-staining cells increased three- to four-fold during pregnancy being maximal at gestational days (GD) 9–12 but were significantly reduced in glucose intolerant mice. Apelin-13 increased β-cell proliferation in isolated mouse islets and INS1E cells, but not glucose-stimulated insulin secretion. Glucose intolerant pregnant mice had significantly elevated serum Apelin levels at GD 9 associated with an increased presence of placental IL-6. Placental expression of the apelinergic axis remained unaltered, however. Results show that the apelinergic system is highly expressed in pancreatic β-cell progenitors and may contribute to β-cell proliferation in pregnancy.

Abstract Background The impact of severe second lockdown measures on older adults’ wellbeing is unknown. We aimed to (i) identify the impact of the second lockdown that resulted from the second wave of COVID-19 cases on older Australians’ quality of life; (ii) compare the impact of second wave lockdowns in Victoria, Australia’s second most populous State, to those in other States and Territories not in lockdown. Methods A national cross-sectional study of community-dwelling older adults completed online questionnaires for quality of life, social networks, healthcare access, and perceived impact of COVID-19 between July to September 2020. Tobit regression was used to measure the relationships of healthcare service access and social networks with quality of life of older adults in Victoria compared to those in the rest of Australia. Results A total of 2,990 respondents (mean [SD] age, 67.3 [7.0]; 66.8 % female) participated. At time of data collection, Victoria’s second COVID-19 lockdown had been in force for an average 51.7 days. Median quality of life scores were significantly higher in Victoria compared to the rest of Australia (t2,827=2.25 p = 0.025). Being female (95 % CI, -0.051–0.020), having lower educational attainment (95 % CI, -0.089–-0.018), receiving government benefits (95 % CI, -0.054–-0.024), having small social networks (95 % CI, 0.006–0.009) and self-reported physical chronic health conditions were all independent predictors of lower quality of life. Conclusions Longer-term studies are required to provide more robust evidence of the impact as restrictions lift and normal social conventions return.

CRISPR-Cas systems provide microbes with adaptive immunity by employing short DNA sequences, termed spacers, that guide Cas proteins to cleave foreign DNA. Class 2 CRISPR-Cas systems are streamlined versions, in which a single RNA-bound Cas protein recognizes and cleaves target sequences. The programmable nature of these minimal systems has enabled researchers to repurpose them into a versatile technology that is broadly revolutionizing biological and clinical research. However, current CRISPR-Cas technologies are based solely on systems from isolated bacteria, leaving the vast majority of enzymes from organisms that have not been cultured untapped. Metagenomics, the sequencing of DNA extracted directly from natural microbial communities, provides access to the genetic material of a huge array of uncultivated organisms. Here, using genome-resolved metagenomics, we identify a number of CRISPR-Cas systems, including the first reported Cas9 in the archaeal domain of life, to our knowledge. This divergent Cas9 protein was found in little-studied nanoarchaea as part of an active CRISPR-Cas system. In bacteria, we discovered two previously unknown systems, CRISPR-CasX and CRISPR-CasY, which are among the most compact systems yet discovered. Notably, all required functional components were identified by metagenomics, enabling validation of robust in vivo RNA-guided DNA interference activity in Escherichia coli. Interrogation of environmental microbial communities combined with in vivo experiments allows us to access an unprecedented diversity of genomes, the content of which will expand the repertoire of microbe-based biotechnologies.

Pleiotrophin (PTN) is a heparin-binding cytokine that is widely expressed during early development and increases in maternal circulation during pregnancy.Aged PTN-deficient mice exhibit insulin resistance, suggesting a role in metabolic control. The objectives of this study were to determine if PTN is expressed in mouse pancreatic β-cells in young vs. adult animals, and its effects on DNA synthesis, β-cell gene expression and glucose-stimulated insulin secretion (GSIS). The Ptn gene was expressed in isolated fractions of young mouse β-cells, especially within immature β-cells with low glucose transporter 2 expression. Expression was retained in the adult pancreas but did not significantly change during pregnancy. PTN and its receptor, phosphotyrosine phosphatase-β/ζ, were also expressed in the proliferative INS1E β-cell line. Fluorescence immunohistochemistry showed that PTN peptide was present in islets of Langerhans in adult mice, associated predominantly with β-cells. The percentage of β-cells staining for PTN did not alter during mouse pregnancy, but intense staining was seen during β-cell regeneration in young mice following depletion of β-cells with streptozotocin. Incubation of INS1E cells with PTN resulted in an increased DNA synthesis as measured by Ki67 localization and increased expression of Pdx1 and insulin. However, both DNA synthesis and GSIS were not altered by PTN in isolated adult mouse islets. The findings show that Ptn is expressed in mouse β-cells in young and adult life and could potentially contribute to adaptive increases in β-cell mass during early life or pregnancy.

IntroductionBehaviour change interventions represent key means for supporting healthy ageing and reducing dementia risk yet brief, scalable behaviour change interventions targeting dementia risk reduction in older adults is currently lacking. Here we describe the aims and design of the three-month Brain Bootcamp initiative that seeks to target multiple dementia risk and protective factors (healthy eating, physical, social and cognitive inactivity), through the use of multiple behaviour change techniques, including goal-setting for behaviour, information about health consequences and physical prompts to change behaviours that reduce dementia risk among older adults. Our secondary aim is to understand participants' views of dementia prevention and explore the acceptability and integration of this campaign into daily life.MethodsBrain Bootcamp is a pre-post feasibility trial conducted in Sydney, Australia beginning in January 2021 until late August. Participants aged ≥65 years living independently in the community (n = 252), recruited through social media and flyers, will provide information about their demographics, medical history, alcohol consumption, smoking habits, mental health, physical activity, cognitive activity, and diet to generate a dementia risk profile at baseline and assess change therein at three-month follow-up. During the intervention, participants will receive a resource pack containing their individual risk profile, educational booklet on dementia risk factors and four physical items designed to prompt physical, social and mental activity, and better nutrition. Outcome measures include change in dementia risk scores, dementia awareness and motivation. A qualitative process evaluation will interview a sample of participants on the acceptability and feasibility of the intervention.DiscussionThis will be the first short-term multi-domain intervention targeting dementia risk reduction in older adults. Findings will generate a new evidence base on how to best support efforts targeting lifestyle changes and to identify ways to optimise acceptability and effectiveness towards brain health for older adults.Trial registration numberACTRN 381046 (registered 17/02/2021); Pre-results.

Eukaryotic Argonaute proteins induce gene silencing by small RNA-guided recognition and cleavage of mRNA targets. Although structural similarities between human and prokaryotic Argonautes are consistent with shared mechanistic properties, sequence and structure-based alignments suggested that Argonautes encoded within CRISPR-cas [clustered regularly interspaced short palindromic repeats (CRISPR)-associated] bacterial immunity operons have divergent activities. We show here that the CRISPR-associated Marinitoga piezophila Argonaute (MpAgo) protein cleaves single-stranded target sequences using 5'-hydroxylated guide RNAs rather than the 5'-phosphorylated guides used by all known Argonautes. The 2.0-Å resolution crystal structure of an MpAgo-RNA complex reveals a guide strand binding site comprising residues that block 5' phosphate interactions. Using structure-based sequence alignment, we were able to identify other putative MpAgo-like proteins, all of which are encoded within CRISPR-cas loci. Taken together, our data suggest the evolution of an Argonaute subclass with noncanonical specificity for a 5'-hydroxylated guide.

Planar polarity describes the coordinated polarization of cells within the plane of a tissue. This is controlled by two main pathways in Drosophila: the Frizzled-dependent core planar polarity pathway and the Fat–Dachsous pathway. Components of both of these pathways become asymmetrically localized within cells in response to long-range upstream cues, and form intercellular complexes that link polarity between neighbouring cells. This review examines if and when the two pathways are coupled, focusing on the Drosophila wing, eye and abdomen. There is strong evidence that the pathways are molecularly coupled in tissues that express a specific isoform of the core protein Prickle, namely Spiny-legs. However, in other contexts, the linkages between the pathways are indirect. We discuss how the two pathways act together and independently to mediate a diverse range of effects on polarization of cell structures and behaviours.

A Goldsmiths based AHRC funded project within the ‘Tackling the Impact of COVID-19’ UKRI call. In collaboration with LASALLE College of the Arts Singapore, Akram Khan Dance Company, and Target3D. In this experimental test session recorded in July 2020, two dancers, one in London, and one in Singapore, are dancing together, but virtually, each wearing an inertial sensor motion capture system. Live and pre-captured dance data was streamed from a dancer in a similar studio space in LASALLE college in Singapore, some 6700 miles away, with barely noticeable delay or latency. Although with occasional technical glitches, including magnetic interference with the suit, this raw footage operates as proof of concept, and a suggestion of what is to come in the next iteration of our research. We position this research practice within a historical and theoretical problematic of networked or ‘distributed performance’ and of telepresence, telematics, and virtuality in dance practice. It is not about recreating the live experience – of ‘being there’ – but rather finding forms of meaningful connection, engaged interest and attention in a digital medium which is decisively and qualitatively different.

BackgroundAccumulating evidence suggests that the COVID-19 pandemic has negatively impacted the mental health of individuals. However, the susceptibility of individuals to be impacted by the pandemic is variable, suggesting potential influences of specific factors related to participants’ demographics, attitudes, and practices. ObjectiveWe aimed to identify the factors associated with psychological symptoms related to the effects of the first wave of the pandemic in a multicountry cohort of internet users. MethodsThis study anonymously screened 13,332 internet users worldwide for acute psychological symptoms related to the COVID-19 pandemic from March 29 to April 14, 2020, during the first wave of the pandemic amidst strict lockdown conditions. A total of 12,817 responses were considered valid. Moreover, 1077 participants from Europe were screened a second time from May 15 to May 30, 2020, to ascertain the presence of psychological effects after the ease down of restrictions. ResultsFemale gender, pre-existing psychiatric conditions, and prior exposure to trauma were identified as notable factors associated with increased psychological symptoms during the first wave of COVID-19 (P

Objective To understand patient burden of treatment of repeated intravitreal injections (IVI) in the management of exudative retinal diseases.Methods and analysis Participants were sampled from a large urban retina specialty practice in Houston, Texas, USA, based on history of ongoing receipt of IVI. The 50-item Questionnaire to Assess Life Impact of Treatment by Intravitreal Injections questionnaire was developed to evaluate the patient experience including discomfort, anxiety, inconvenience and satisfaction. Categorial principal components analysis (CATPCA) was performed to assess construct validity and internal consistency. A subset of these items was used to establish a measure of total treatment burden, referred to as the IVI Treatment Burden Score (TBS).Results 142 patients participated in this study. CATPCA analysis revealed five dimensions of patient burden: disruption of normal routine or capacity, anxiety, frequency of visits, chronicity of disease and perceived treatment value or satisfaction. Together, these dimensions accounted for 67% of variance explained. Cronbach’s alpha was 0.97. The most frequently cited cause of discomfort was the feeling after anaesthetic wore off. The most common source of anxiety was fear of injection and associated discomfort or pain. Regarding inconvenience, patients reported temporary postinjection debilitation, requiring an average of 8 hours for recovery per treatment. The most frequently identified sources of satisfaction were confidence in the provider or treatment and interactions with staff.Conclusions Understanding and quantifying the patient burden associated with repeated IVI for exudative retinal diseases can reveal opportunities to improve delivery methods. The TBS could serve to inform strategies to maximise treatment adherence and optimise patient experiences.

The core planar polarity proteins are essential mediators of tissue morphogenesis, controlling both the polarised production of cellular structures and polarised tissue movements. During development the core proteins promote planar polarisation by becoming asymmetrically localised to opposite cell edges within epithelial tissues, forming intercellular protein complexes that coordinate polarity between adjacent cells. Here we describe a novel protein complex that regulates the asymmetric localisation of the core proteins in the Drosophila pupal wing. DAnkrd49 (an ankyrin repeat protein) and Bride of Doubletime (Bdbt, a non-canonical FK506 binding protein family member) physically interact, and regulate each other's levels in vivo. Loss of either protein results in a reduction in core protein asymmetry and disruption of the placement of trichomes at the distal edge of pupal wing cells. Post-translational modifications are thought to be important for the regulation of core protein behaviour and their sorting to opposite cell edges. Consistent with this, we find that loss of DAnkrd49 or Bdbt leads to reduced phosphorylation of the core protein Dishevelled and to decreased Dishevelled levels both at cell junctions and in the cytoplasm. Bdbt has previously been shown to regulate activity of the kinase Discs Overgrown (Dco, also known as Doubletime or Casein Kinase Iε), and Dco itself has been implicated in regulating planar polarity by phosphorylating Dsh as well as the core protein Strabismus. We demonstrate that DAnkrd49 and Bdbt act as dominant suppressors of Dco activity. These findings support a model whereby Bdbt and DAnkrd49 act together to modulate the activity of Dco during planar polarity establishment.