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Abstract Focal adhesions form liquid-like assemblies around activated integrin receptors at the plasma membrane. How they achieve their flexible properties is not well understood. Here, we use recombinant focal adhesion proteins to reconstitute the core structural machinery in vitro. We observe liquid-liquid phase separation of the core focal adhesion proteins talin and vinculin for a spectrum of conditions and interaction partners. Intriguingly, we show that binding to PI(4,5)P2-containing membranes triggers phase separation of these proteins on the membrane surface, which in turn induces the enrichment of integrin in the clusters. We suggest a mechanism by which 2-dimensional biomolecular condensates assemble on membranes from soluble proteins in the cytoplasm: lipid-binding triggers protein activation and thus, liquid-liquid phase separation of these membrane-bound proteins. This could explain how early focal adhesions maintain a structured and force-resistant organization into the cytoplasm, while still being highly dynamic and able to quickly assemble and disassemble.

Problem-solving abilities, creative and critical thinking, communication skills, and teamwork are now recognized as fundamental determinants of professional success, especially in vocational professions, such as veterinary science. Tertiary education is now obliged to provide opportunities for students to become proficient in these qualities. With this in mind, the principal author, an equine science senior lecturer, attempted to increase student engagement by developing a new active learning, student-centered one-hour teaching episode on 'equine diarrhea', to replace the traditional didactic lecture format. The aim of the study, therefore, was to share the principal author's journey in the development and implementation of this active learning episode and to explain why it represents a simple but effective method of promoting student engagement. In addition, the adaptation of this method into an online teaching and learning format is briefly discussed. The effectiveness of this active learning method is also explored by comparing it with the traditional didactic method of delivery. Students in the active learning class reported that the learning activity had enhanced their skills in clinical reasoning, problem-solving, and communication. They also described themselves as active participants in the learning process. Students in the traditional didactic class reported that they were satisfied with the time allocated to the lecture, felt that the lecture was well organized and managed, and subsequently felt adequately prepared to answer conventional examination questions relating to equine diarrhea. Other issues, such as student resistance to changes in teaching formats, the effectiveness of group work or teamwork, levels of student confidence within an active learning environment, and the importance of high-quality facilitation during active learning activities, are also discussed in this paper. Although the active learning method described is not novel, the authors hope that fellow educators, across any tertiary discipline, might find that the method described represents a quick and simple method of transforming a single didactic lecture into an enjoyable and engaging learning activity.

Parkinson's disease (PD) and Dementia with Lewy bodies (DLB) are characterized by neuronal α-synuclein (α-syn) inclusions termed Lewy Pathology, which are abundant in the amygdala. The basolateral amygdala (BLA), in particular, receives projections from the thalamus and cortex. These projections play a role in cognition and emotional processing, behaviors which are impaired in α-synucleinopathies. To understand if and how pathologic α-syn impacts the BLA requires animal models of α-syn aggregation. Injection of α-syn pre-formed fibrils (PFFs) into the striatum induces robust α-syn aggregation in excitatory neurons in the BLA that corresponds with reduced contextual fear conditioning. At early time points after aggregate formation, cortico-amygdala excitatory transmission is abolished. The goal of this project was to determine if α-syn inclusions in the BLA induce synaptic degeneration and/or morphological changes. In this study, we used C57BL/6 J mice injected bilaterally with PFFs in the dorsal striatum to induce α-syn aggregate formation in the BLA. A method was developed using immunofluorescence and three-dimensional reconstruction to analyze excitatory cortico-amygdala and thalamo-amygdala presynaptic terminals closely juxtaposed to postsynaptic densities. The abundance and morphology of synapses were analyzed at 6- or 12-weeks post-injection of PFFs. α-Syn aggregate formation in the BLA did not cause a significant loss of synapses, but cortico-amygdala and thalamo-amygdala presynaptic terminals and postsynaptic densities with aggregates of α-syn show increased volumes, similar to previous findings in human DLB cortex, and in non-human primate models of PD. Transmission electron microscopy showed that asymmetric synapses in mice with PFF-induced α-syn aggregates have reduced synaptic vesicle intervesicular distances, similar to a recent study showing phospho-serine-129 α-syn increases synaptic vesicle clustering. Thus, pathologic α-syn causes major alterations to synaptic architecture in the BLA, potentially contributing to behavioral impairment and amygdala dysfunction observed in synucleinopathies.

Abstract Progress in the field of ecological stoichiometry has demonstrated that the outcome of ecological interactions can often be predicted a priori based on the nutrient ratios (e.g., carbon: nitrogen: phosphorus, C:N:P) of interacting organisms. However, the challenges of accurately measuring the nutrient content of active parasites within hosts has limited our ability to rigorously apply ecological stoichiometry to host–parasite systems. Traditional nutrient analyses require high parasite biomasses, often preventing individual‐level analyses. This prevents researchers from estimating variation in the nutrient content of individual parasites within a single host infrapopulation, a critical factor that could define how the ecology of the parasite affects the host–parasite interaction. Here, we explain how energy dispersive technology, a technique currently used to measure the elemental content of free‐living microbes, can be adapted for parasitic microbial infrapopulations. We demonstrate the power of accurately quantifying the biomass stoichiometry of individual microbial parasites sampled directly from individual hosts. Using this approach, we show that the stoichiometric composition of two microbial parasites capable of infecting the same host are stoichiometrically distinct and respond to host diet quality differently. We also demonstrate that characteristics of the stoichiometric trait distributions of these infrapopulations were important predictors of host fecundity, a proxy for virulence in this system, and better predictors of parasite load than the mean parasite stoichiometry or our parasite and diet treatments alone. EDS provides a rigorous tool for applying ecological stoichiometry to host–parasite systems and enables researchers to explore the nutritional physiology of host–parasite interactions at a scale that is more relevant to the ecology and evolution of the system than traditional nutrient analyses. Here we demonstrate that this level of resolution provides useful insights into the diet‐dependent physiology of microbial parasites and their hosts. We anticipate that this improved level of resolution has the potential to elucidate a range of eco–evo interactions in host–parasite systems that were previously unobservable.

Drosophila is a powerful model to study how lipids affect spermatogenesis. Yet, the contribution of neutral lipids, a major lipid group which resides in organelles called lipid droplets (LD), to sperm development is largely unknown. Emerging evidence suggests LD are present in the testis and that loss of neutral lipid- and LD-associated genes causes subfertility; however, key regulators of testis neutral lipids and LD remain unclear. Here, we show LD are present in early-stage somatic and germline cells within the Drosophila testis. We identified a role for triglyceride lipase brummer (bmm) in regulating testis LD, and found that whole-body loss of bmm leads to defects in sperm development. Importantly, these represent cell-autonomous roles for bmm in regulating testis LD and spermatogenesis. Because lipidomic analysis of bmm mutants revealed excess triglyceride accumulation, and spermatogenic defects in bmm mutants were rescued by genetically blocking triglyceride synthesis, our data suggest that bmm-mediated regulation of triglyceride influences sperm development. This identifies triglyceride as an important neutral lipid that contributes to Drosophila sperm development, and reveals a key role for bmm in regulating testis triglyceride levels during spermatogenesis.

Abstract The nutrient content of host resources can influence the abundance of parasites within an ecosystem, but linking specific nutrients in a host to the abundance of different parasite taxa remains a challenge. Here, we work to forge this link by quantifying the relationship between the nutrient content of specific infection sites and the abundance of multiple parasite taxa within the digestive tract of largemouth bass (Micropterus salmoides) collected from the Mississippi River. To generate a mechanistic understanding of these relationships, we tested four basic predictions: (1) the nutrient content of different host tissues (infection sites) varies within and across hosts, (2) the nutrient content of parasite genera differs from that of their host tissue(s), (3) the nutrient content of parasite genera differ from one another and (4) the nutrient content of host tissues is related to the nutrient content and abundance of parasite genera. We found support for each of these predictions. We found stoichiometric differences between the digestive tissues we examined. We also found that across hosts, intestine and pyloric caeca C:N ratios increased and %N decreased with fish condition factor. Both of the actively feeding parasitic genera we measured had lower C:N ratios compared to both their host tissue and other encysted/non‐reproductive genera, suggesting the potential for N limitation of these parasites in the intestines or pyloric caeca of hosts. Consistent with this possibility, we found that the total number of actively feeding parasitic worms in the pyloric caeca increased with that tissue's N:P ratio (but was not related to host condition factor). Our results suggest that parasites encounter significant variation in nutrient content within and across hosts and that this variation may influence the abundance of actively feeding parasites. This work highlights the need for additional empirical comparisons of parasite stoichiometry across tissues and individual hosts.

Abstract Objective How we express and describe emotion is shaped by sociocultural norms. These sociocultural norms may also affect emotional self-awareness, i.e., how we identify and make sense of our own emotions. Previous studies have found lower emotional self-awareness in East Asian compared to Western samples using self-report measures. However, studies using behavioural methods did not provide clear evidence of reduced emotional self-awareness in East Asian groups. This may be due to different measurement tools capturing different facets of emotional self-awareness. Results To investigate this issue further, we compared the emotional self-awareness of Japanese (n = 29) and United Kingdom (UK) (n = 43) adults using the self-report Toronto Alexithymia Scale (TAS-20), alongside two behavioural measures – the Emotional Consistency Task (EC-Task) and the Photo Emotion Differentiation Task (PED-Task). Japanese adults showed higher TAS-20 scores than UK participants, indicating greater self-reported difficulties with emotional self-awareness. Japanese participants also had lower EC-Task scores than UK adults, indicating a lower ability to differentiate between levels of emotional intensity. PED-Task performance did not show clear group differences. These findings suggest that cross-cultural differences in emotional self-awareness vary with the task used, because different tasks assess distinct aspects of this ability. Future research should attempt to capture these different aspects of emotional self-awareness.

Chimeric antigen receptor (CAR) T cell therapy has revolutionized the management of relapsed and refractory myeloma, with excellent outcomes and a tolerable safety profile. High dose chemotherapy with autologous hematopoietic stem cell transplantation (AHCT) is established as a mainstream of newly diagnosed multiple myeloma (NDMM) management in patients who are young and fit enough to tolerate such intensity. This standard was developed based on randomized trials comparing AHCT to chemotherapy in the era prior to novel agents. More recently, larger studies have primarily shown a progression free survival (PFS) benefit of upfront AHCT, rather than overall survival (OS) benefit. There is debate about the significance of this lack of OS, acknowledging the potential confounders of the chronic nature of the disease, study design and competing harms and benefits of exposure to AHCT. Indeed upfront AHCT may not be as uniquely beneficial as we once thought, and is not without risk. New quadruple-agent regimens are highly active and effective in achieving a deep response as quantified by measurable residual disease (MRD). The high dose chemotherapy administered with AHCT imposes a burden of short and long-term adverse effects, which may alter the disease course and patient’s ability to tolerate future therapies. Some high-risk subgroups may have a more valuable benefit from AHCT, though still ultimately suffer poor outcomes. When compared to the outcomes of CAR T cell therapy, the question of whether AHCT can or indeed should be deferred has become an important topic in the field. Deferring AHCT may be a personalized decision in patients who achieve MRD negativity, which is now well established as a key prognostic factor for PFS and OS. Reserving or re-administering AHCT at relapse is feasible in many cases and holds the promise of resetting the T cell compartment and opening up options for immune reengagement. It is likely that personalized MRD-guided decision making will shape how we sequence in the future, though more studies are required to delineate when this is safe and appropriate.

We present a realization of highly frustrated planar triangular antiferromagnetism achieved in a quasi-three-dimensional artificial spin system consisting of monodomain Ising-type nanomagnets lithographically arranged onto a deep-etched silicon substrate. We demonstrate how the three-dimensional spin architecture results in the first direct observation of long-range ordered planar triangular antiferromagnetism, in addition to a highly disordered phase with short-range correlations, once competing interactions are perfectly tuned. Our work demonstrates how escaping two-dimensional restrictions can lead to new types of magnetically frustrated metamaterials.

Five years after the last prostatic carcinoma grading consensus conference of the International Society of Urological Pathology (ISUP), accrual of new data and modification of clinical practice require an update of current pathologic grading guidelines. This manuscript summarizes the proceedings of the ISUP consensus meeting for grading of prostatic carcinoma held in September 2019, in Nice, France. Topics brought to consensus included the following: (1) approaches to reporting of Gleason patterns 4 and 5 quantities, and minor/tertiary patterns, (2) an agreement to report the presence of invasive cribriform carcinoma, (3) an agreement to incorporate intraductal carcinoma into grading, and (4) individual versus aggregate grading of systematic and multiparametric magnetic resonance imaging-targeted biopsies. Finally, developments in the field of artificial intelligence in the grading of prostatic carcinoma and future research perspectives were discussed.

Phytophthora species are oomycete plant pathogens that cause great economic and ecological impacts. The Phytophthora genus includes over 180 known species, infecting a wide range of plant hosts, including crops, trees, and ornamentals. We sequenced the genomes of 31 individual Phytophthora species and 24 individual transcriptomes to study genetic relationships across the genus. De novo genome assemblies revealed variation in genome sizes, numbers of predicted genes, and in repetitive element content across the Phytophthora genus. A genus-wide comparison evaluated orthologous groups of genes. Predicted effector gene counts varied across Phytophthora species by effector family, genome size, and plant host range. Predicted numbers of apoplastic effectors increased as the host range of Phytophthora species increased. Predicted numbers of cytoplasmic effectors also increased with host range but leveled off or decreased in Phytophthora species that have enormous host ranges. With extensive sequencing across the Phytophthora genus, we now have the genomic resources to evaluate horizontal gene transfer events across the oomycetes. Using a machine-learning approach to identify horizontally transferred genes with bacterial or fungal origin, we identified 44 candidates over 36 Phytophthora species genomes. Phylogenetic reconstruction indicates that the transfers of most of these 44 candidates happened in parallel to major advances in the evolution of the oomycetes and Phytophthora spp. We conclude that the 31 genomes presented here are essential for investigating genus-wide genomic associations in genus Phytophthora. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Background European canker, caused by the fungal pathogen Neonectria ditissima, is an economically damaging disease in apple producing regions of the world – especially in areas with moderate temperatures and high rainfall. The pathogen has a wide host range of hardwood perennial species, causing trunk cankers, dieback and branch lesions in its hosts. Although apple scion germplasm carrying partial resistance to the disease has been described, little is still known of the genetic basis for this quantitative resistance. Results Resistance to Neonectria ditissima was studied in a multiparental population of apple scions using several phenotyping methods. The studied population consists of individuals from multiple families connected through a common pedigree. The degree of disease of each individual in the population was assessed in three experiments: artificial inoculations of detached dormant shoots, potted trees in a glasshouse and in a replicated field experiment. The genetic basis of the differences in disease was studied using a pedigree-based analysis (PBA). Three quantitative trait loci (QTL), on linkage groups (LG) 6, 8 and 10 were identified in more than one of the phenotyping strategies. An additional four QTL, on LG 2, 5, 15 and 16 were only identified in the field experiment. The QTL on LG2 and 16 were further validated in a biparental population. QTL effect sizes were small to moderate with 4.3 to 19% of variance explained by a single QTL. A subsequent analysis of QTL haplotypes revealed a dynamic response to this disease, in which the estimated effect of a haplotype varied over the field time-points. Conclusions This study describes the first identified QTL associated with resistance to N. ditissima in apple scion germplasm. The results from this study show that QTL present in germplasm commonly used in apple breeding have a low to medium effect on resistance to N. ditissima. Hence, multiple QTL will need to be considered to improve resistance through breeding.

Abstract Background Employment disruptions can impact smoking and alcohol consumption. During the COVID-19 pandemic, many countries implemented furlough schemes to prevent job loss. We examine how furlough was associated with smoking, vaping and alcohol consumption in the UK. Methods Data from 27,841 participants in eight UK adult longitudinal surveys were analysed. Participants self-reported employment status and current smoking, current vaping and alcohol consumption (>4 days/week or 5+ drinks per typical occasion) both before and during the early stages of the pandemic (April–July 2020). Risk ratios were estimated within each study using modified Poisson regression, adjusting for a range of potential confounders, including pre-pandemic behaviour. Findings were synthesised using random effects meta-analysis. Results Compared to stable employment and after adjustment for pre-pandemic characteristics, furlough was not associated with smoking (ARR = 1.05; 95% CI: 0.95–1.16; I 2: 10%), vaping (ARR = 0.89; 95% CI: 0.74–1.08; I 2: 0%) or drinking (ARR = 1.03; 95% CI: 0.94–1.13; I 2: 48%). There were similar findings for no longer being employed, and stable unemployment, though this varied by sex: stable unemployment was associated with smoking for women (ARR = 1.35; 95% CI: 1.00–1.82; I 2: 47%) but not men (0.84; 95% CI: 0.67–1.05; I 2: 0%). No longer being employed was associated with vaping among women (ARR = 2.74; 95% CI: 1.59–4.72; I 2: 0%) but not men (ARR = 1.25; 95% CI: 0.83–1.87; I 2: 0%). Conclusions We found no clear evidence of furlough or unemployment having adverse impacts on smoking, vaping or drinking behaviours during the early stages of the COVID-19 pandemic in the UK. Differences in risk compared to those who remained employed were largely explained by pre-pandemic characteristics.

Emotional self-awareness is increasingly suggested to be an area of difficulty in autism that may predict socioemotional outcomes for this population. However, whether emotional self-awareness is consistently diminished in autism across age and methodology remains unclear. We systematically reviewed 47 papers measuring emotional self-awareness in autistic (n=1387) and non-autistic (n=1433) participants. Most studies relied on self-report. Of studies testing for group differences, the majority (32/41) found significantly poorer emotional self-awareness in autism. Meta-analyses of self-report measures found that emotional self-awareness was significantly poorer in autism (d=1.16). However, when examining age groups individually, autistic children of age 12 years and under were not significantly different from their peers (d=0.03). Instead, difficulties emerged during adolescence (d=0.63) and increased with age (d=1.16-1.58). The pattern of emotional self-awareness difficulties being more common in autism, and worsening with age is similar to the development of mental health difficulties in autism. However, findings rely on self-perception and so may reflect poor self-beliefs of socioemotional competence. We propose that negative self-beliefs in autistic populations may account for findings of low emotional self-awareness.

Magnetic monopoles, proposed as elementary particles that act as isolated magnetic south and north poles, have long attracted research interest as magnetic analogs to electric charge. In solid-state physics, a classical analog to these elusive particles has emerged as topological excitations within pyrochlore spin ice systems. We present the first real-time imaging of emergent magnetic monopole motion in a macroscopically degenerate artificial spin ice system consisting of thermally activated Ising-type nanomagnets lithographically arranged onto a pre-etched silicon substrate. A real-space characterization of emergent magnetic monopoles within the framework of Debye-Hückel theory is performed, providing visual evidence that these topological defects act like a plasma of Coulomb-type magnetic charges. In contrast to vertex defects in a purely two-dimensional artificial square ice, magnetic monopoles are free to evolve within a divergence-free vacuum, a magnetic Coulomb phase, for which features in the form of pinch-point singularities in magnetic structure factors are observed.

Current understanding of Long COVID is limited, in part, due to lack of evidence from population-representative studies. Here, the authors analyse data from ten UK population-based studies and electronic health records, and find wide variation in the frequency of Long COVID between studies but some consistent risk factors.