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Abstract The chiral separation capability of Chiral-CD-Ph column, containing phenylcarbamate-β-cyclodextrin as the chiral selector in polar organic mode was investigated. A total of twenty-five compounds with different structures and acid–base properties were evaluated, and twenty of them were separated using acetonitrile or methanol as eluent. The effects of various chromatographic parameters, such as the type and proportion of organic modifier, flow rate, and column temperature were analyzed in detail in relation to chromatographic performance. A U-shape retention curve was observed when a mixture of acetonitrile and methanol was used as the eluent, indicating different types of interactions in different solvent mixtures. Van 't Hoff analysis was used for calculation of thermodynamic parameters which revealed that the enantioseparation is mainly enthalpy controlled; however, entropic control was also observed. The enantiomer recognition ability at the atomic level was also investigated through a molecular docking study, which revealed surface binding in polar organic mode instead of inclusion complexation. Our work proves that the phenylcarbamate-β-cyclodextrin-based chiral stationary phase can be effectively used in polar organic mode for the chiral separation of structurally diverse compounds. Furthermore, it is important to note that our study demonstrated that surface binding is responsible for the formation of supramolecular complexes in certain cyclodextrin derivatives.

Abstract Background In the last three decades there is a growing recognition in the dental profession that dental education must go beyond teaching the technicalities of dentistry and include professionalism and communication skills that the future dentist may need. Such skills are best taught in a student-centered way. Literature suggests that student-centered elements are difficult to introduce in traditional, teacher-centered curricula. This is especially true in post-communist countries where higher education was under strict state control for decades. The aim of the piece of research presented here was to investigate how difficult it is to introduce a student-centered career skills course in a traditionally teacher-centered dental curriculum. Methods Considering the needs of our final-year dental students and Super’s model of career development, we created an undergraduate curricular career skills course running for two semesters in two languages (Hungarian and English). The primary aim of the course is to help students with their career expectations and develop their identity as a professional workforce. The secondary aim is to teach skills that students can use when applying for a job. At the end of the semesters, we assessed our students’ satisfaction with various aspects of the course by using a questionnaire. Results were analyzed item-wise and according to the main aspects of the course (i.e., groups of items organized around a particular aspect). Results General satisfaction with the course was high, and practical skills training (such as CV and motivation letter writing) got the highest scores. From the answers it appears that the students were the least comfortable with having to deal with their personal values and preferences. Conclusions While it is common for universities to offer various forms of career intervention, to our knowledge, no other university offers a curricular career skills course specifically for dental students. Our student-centered course designed in a problem-based learning framework worked even in a traditionally teacher-centered educational environment, where university students are rarely encouraged to be active participants in courses. By sharing our experience, we would like to encourage our fellow dental educators working in similar environments to devise and offer such courses.

Dental turbines and scalers, used every day in dental operatories, feature built-in water spray that generates considerable amounts of water aerosol. The problem is that it is not exactly known how much. Since the outbreak of COVID-19, several aerosol safety recommendations have been issued-based on little empirical evidence, as almost no data are available on the exact aerosol concentrations generated during dental treatment. Similarly, little is known about the differences in the efficacy of different commercially available aerosol control systems to reduce in-treatment aerosol load. In this in vitro study, we used spectrometry to explore these questions. The time-dependent effect of conventional airing on aerosol concentrations was also studied. Everyday patient treatment situations were modeled. The test setups were defined by the applied instrument and its spray direction (high-speed turbine with direct/indirect airspray or ultrasonic scaler with indirect airspray) and the applied aerosol control system (the conventional high-volume evacuator or a lately introduced aerosol exhaustor). Two parameters were analyzed: total number concentration in the entire measurement range of the spectrometer and total number concentration within the 60 to 384 nm range. The results suggest that instrument type and spray direction significantly influence the resulting aerosol concentrations. Aerosol generation by the ultrasonic scaler is easily controlled. As for the high-speed turbine, the efficiency of control might depend on how exactly the instrument is used during a treatment. The results suggest that scenarios where the airspray is frequently directed toward the air of the operatory are the most difficult to control. The tested control systems did not differ in their efficiency, but the study could not provide conclusive results in this respect. With conventional airing through windows with a standard fan, a safety airing period of at least 15 minutes between treatments is recommended.

BACKGROUND: Population-based studies have identified smoking as a pathogenetic factor in chronic periodontitis. At the same time, chronic periodontal disease has also been found to occur more often in persons suffering from psoriasis than in controls with no psoriasis. It is known that smoking aggravates both periodontal disease and psoriasis, but so far it has not been investigated how smoking influences the occurrence and severity of periodontal disease in psoriasis. METHODS: A hospital-based study was conducted to investigate this question. The study population consisted of 82 psoriasis patients and 89 controls. All patients received a full-mouth periodontal examination, and a published classification based on bleeding on probing, clinical attachment level and probing depth was utilized for staging. Both patients and controls were divided into smoker and non-smoker groups, and the resulting groups were compared in terms of periodontal status. Beyond the descriptive statistics, odds ratios were computed. RESULTS: Psoriasis in itself increased the likelihood of severe periodontal disease to 4.373 (OR, as compared to non-smoker controls, p

Abstract Functional magnetic resonance imaging (fMRI) has dramatically advanced non-invasive human brain mapping and decoding. Functional near-infrared spectroscopy (fNIRS) and high-density diffuse optical tomography (HD-DOT) non-invasively measure blood oxygen fluctuations related to brain activity, like fMRI, at the brain surface, using more-lightweight equipment that circumvents ergonomic and logistical limitations of fMRI. HD-DOT grids have smaller inter-optode spacing (~ 13 mm) than sparse fNIRS (~ 30 mm) and therefore provide higher image quality, with spatial resolution ~ 1/2 that of fMRI, when using the several source-detector distances (13–40 mm) afforded by the HD-DOT grid. Herein, simulations indicated reducing inter-optode spacing to 6.5 mm, creating a higher-density grid with more source-detector distances, would further improve image quality and noise-resolution tradeoff, with diminishing returns below 6.5 mm. We then constructed an ultra-high-density DOT system (6.5-mm spacing) with 140 dB dynamic range that imaged stimulus-evoked activations with 30–50% higher spatial resolution and repeatable multi-focal activity with excellent agreement with participant-matched fMRI. Further, this system decoded visual stimulus position with 19–35% lower error than previous HD-DOT, throughout occipital cortex.

Abstract Background The concept of children’s rights emerged during the 1980s and emphasised the role of children as active participants in matters which concern them. Aim This review aims to identify and synthesise the empirical evidence base on healthcare professionals’ (HCPs) understanding of children’s rights. Methods Five electronic databases (PubMed, CINAHL, Embase, PsycINFO, and the Web of Science) were systematically searched in May 2023. The Mixed Methods Appraisal Tool (MMAT) was used to quality appraise full-text papers included in the review. A descriptive narrative synthesis of the studies’ findings was performed. Results A total of 15 relevant studies from 10 countries were identified and included in the review. The number of participants included ranged from 6 to 1048 for HCPs with a broad range of sampling methods. Based on the narrative synthesis of the included studies, three main themes were identified: (1) Barriers to implementing children’s rights in healthcare, (2) Factors that contribute to children’s rights implementation, and (3) Study instruments used to measure outcomes. Conclusions HCPs require a better understanding of children’s rights to implement these rights into practice. Listening to children, building trusting relationships with children, and continuing professional development of HCPs could help to address barriers to understanding children’s rights. There is a pressing need for the development of a tool that is capable of tracking changes in the understanding of children’s rights in healthcare environments as efforts to increase awareness become more widely recognised.

Abstract Emerging infectious diseases are of major concern to animal and human health. Recent emergence of high pathogenicity avian influenza virus (HPAIV) (H5N1 clade 2.3.4.4b) led to substantial global mortality across a range of host species. Co-occurring species showed marked differences in mortality, generating an urgent need for better epidemiological understanding within affected populations. We therefore tested for antibodies, indicative of previous exposure and recovery, and for active viral infection in apparently healthy individuals (n = 350) across five co-occurring seabird species on the Isle of May, Scotland, during 2023, following H5N1 HPAIV associated mortality in the preceding summer. Antibody prevalence to AIV subtypes varied substantially between species, ranging from 1.1% in European shags (Gulosus aristotelis) (to H5) to 78.7% in black-legged kittiwakes (Rissa tridactyla) (to H16 or both H13 and H16), and between 31 and 41% for three auk species (H5, H16 or both). At least 20.4% of auks had antibodies to an as yet unidentified subtype, suggesting further subtypes circulating in the population. We found low levels of active, but asymptomatic, AIV infection in individuals (1.6–4.5%), but excluded this as H5N1. Our results emphasise the importance of testing healthy individuals to understand the prevalence of co-circulating AIV subtypes in wild populations, and the potential for future reassortment events which could alter virus behaviour and impact.

Abstract One of the hallmarks of cancer is metabolic reprogramming which controls cellular homeostasis and therapy resistance. Here, we investigated the effect of momordicine-I (M-I), a key bioactive compound from Momordica charantia (bitter melon), on metabolic pathways in human head and neck cancer (HNC) cells and a mouse HNC tumorigenicity model. We found that M-I treatment on HNC cells significantly reduced the expression of key glycolytic molecules, SLC2A1 (GLUT-1), HK1, PFKP, PDK3, PKM, and LDHA at the mRNA and protein levels. We further observed reduced lactate accumulation, suggesting glycolysis was perturbed in M-I treated HNC cells. Metabolomic analyses confirmed a marked reduction in glycolytic and TCA cycle metabolites in M-I-treated cells. M-I treatment significantly downregulated mRNA and protein expression of essential enzymes involved in de novo lipogenesis, including ACLY, ACC1, FASN, SREBP1, and SCD1. Using shotgun lipidomics, we found a significant increase in lysophosphatidylcholine and phosphatidylcholine loss in M-I treated cells. Subsequently, we observed dysregulation of mitochondrial membrane potential and significant reduction of mitochondrial oxygen consumption after M-I treatment. We further observed M-I treatment induced autophagy, activated AMPK and inhibited mTOR and Akt signaling pathways and leading to apoptosis. However, blocking autophagy did not rescue the M-I-mediated alterations in lipogenesis, suggesting an independent mechanism of action. M-I treated mouse HNC MOC2 cell tumors displayed reduced Hk1, Pdk3, Fasn, and Acly expression. In conclusion, our study revealed that M-I inhibits glycolysis, lipid metabolism, induces autophagy in HNC cells and reduces tumor volume in mice. Therefore, M-I-mediated metabolic reprogramming of HNC has the potential for important therapeutic implications. Graphical Abstract

Abstract Light-based gamma entrainment using sensory stimuli (GENUS) shows considerable potential for the treatment of Alzheimer’s disease (AD) in both animal and human models. While the clinical efficacy of GENUS for AD is paramount, its effectiveness will eventually also rely on the barrier to treatment adherence imposed by the discomfort of gazing at luminance flickering (LF) light. Currently, there have been few attempts to improve the comfort of GENUS. Here we investigate if Invisible spectral flicker (ISF), a novel type of light-based 40 Hz GENUS for which the flicker is almost imperceptible, can be used as a more comfortable option. We found that whereas ISF, LF, and chromatic flicker (CF) all produce a 40 Hz steady-state visually evoked potential (SSVEP), ISF scores significantly better on measures of comfort and perceived flicker. We also demonstrate that, while there is a trend towards a lower SSVEP response, reducing the stimulation brightness has no significant effect on the 40 Hz SSVEP or perceived flicker, though it significantly improves comfort. Finally, there is a slight decrease in the 40 Hz SSVEP response when stimulating with ISF from increasingly peripheral angles. This may ease the discomfort of GENUS treatment by freeing patients from gazing directly at the light.

Abstract The issue of adolescent secondhand smoke (SHS) is globally significant, given that it serves as a preventable risk factor for disease prevalence and mortality rates among youth. This study evaluates trends in adolescent SHS across 27 countries from 2003 to 2021, aiming to identify global variations and sex-specific differences, providing insights for future policy recommendations. The data for this study were collected from the Global School-based Student Health Survey conducted by the World Health Organization, 2003–2021. The term SHS in this context denoted how many days people smoked in their presence for at least one day within the past 7 days. Linear regression models were employed to examine the trends of SHS exposure by country. In the present analysis based on 175,370 adolescents (male, 46.69%) aged between 13 and 15 years from 27 countries across the five continents, SHS decreased in 14 countries but exhibited an increasing trend in 3 countries (Benin [β, 3.20; 95% CI, 2.50–3.91]; Maldives [β, 0.93; 0.21–1.65]; Myanmar [β, 0.62; 0.29–0.95]). Significant increasing trends in females were observed in the Maldives (β, 1.18; 0.25–2.11) and Thailand (β, 0.36; 0.04–0.69) while males showed increasing trends in Kuwait (β, 1.90; 0.79-3.00) and Vanuatu (β, 1.95; 0.35–3.54). Our analysis of adolescents from 27 countries indicates that the majority of nations exhibited decreasing trends in SHS. This suggests that global efforts to prevent SHS are achievable through individual initiatives and international monitoring. However, the presence of increasing trends in certain countries underscores the need for stronger regulations and policies to address this issue.

Abstract Calcium imaging allows recording from hundreds of neurons in vivo with the ability to resolve single cell activity. Evaluating and analyzing neuronal responses, while also considering all dimensions of the data set to make specific conclusions, is extremely difficult. Often, descriptive statistics are used to analyze these forms of data. These analyses, however, remove variance by averaging the responses of single neurons across recording sessions, or across combinations of neurons, to create single quantitative metrics, losing the temporal dynamics of neuronal activity, and their responses relative to each other. Dimensionally Reduction (DR) methods serve as a good foundation for these analyses because they reduce the dimensions of the data into components, while still maintaining the variance. Nonnegative Matrix Factorization (NMF) is an especially promising DR analysis method for analyzing activity recorded in calcium imaging because of its mathematical constraints, which include positivity and linearity. We adapt NMF for our analyses and compare its performance to alternative dimensionality reduction methods on both artificial and in vivo data. We find that NMF is well-suited for analyzing calcium imaging recordings, accurately capturing the underlying dynamics of the data, and outperforming alternative methods in common use.

Abstract Background Neoantigen vaccines can induce or enhance highly specific antitumor immune responses with minimal risk of autoimmunity. We have developed a neoantigen DNA vaccine platform capable of efficiently presenting both HLA class I and II epitopes and performed a phase 1 clinical trial in triple-negative breast cancer patients with persistent disease on surgical pathology following neoadjuvant chemotherapy, a patient population at high risk of disease recurrence. Methods Expressed somatic mutations were identified by tumor/normal exome sequencing and tumor RNA sequencing. The pVACtools software suite of neoantigen prediction algorithms was used to identify and prioritize cancer neoantigens and facilitate vaccine design for manufacture in an academic GMP facility. Neoantigen DNA vaccines were administered via electroporation in the adjuvant setting (i.e., following surgical removal of the primary tumor and completion of standard of care therapy). Vaccines were monitored for safety and immune responses via ELISpot, intracellular cytokine production via flow cytometry, and TCR sequencing. Results Eighteen subjects received three doses of a neoantigen DNA vaccine encoding on average 11 neoantigens per patient (range 4–20). The vaccinations were well tolerated with relatively few adverse events. Neoantigen-specific T cell responses were induced in 14/18 patients as measured by ELISpot and flow cytometry. At a median follow-up of 36 months, recurrence-free survival was 87.5% (95% CI: 72.7–100%) in the cohort of vaccinated patients. Conclusion Our study demonstrates neoantigen DNA vaccines are safe, feasible, and capable of inducing neoantigen-specific immune responses. Clinical trial registration number NCT02348320.

Abstract Spinal projection neurons (PNs) are defined by long axons that travel from their origin in the spinal cord to the brain where they relay sensory information from the body. The existence and function of a substantial axon collateral network, also arising from PNs and remaining within the spinal cord, is less well appreciated. Here we use a retrograde viral transduction strategy to characterise a novel subpopulation of deep dorsal horn spinoparabrachial neurons. Brainbow assisted analysis confirmed that virally labelled PN cell bodies formed a discrete cell column in the lateral part of Lamina V (LVlat) and the adjoining white matter. These PNs exhibited large dendritic territories biased to regions lateral and ventral to the cell body column and extending considerable rostrocaudal distances. Optogenetic activation of LVLat PNs confirmed this population mediates widespread signalling within spinal cord circuits, including activation in the superficial dorsal horn. This signalling was also demonstrated with patch clamp recordings during LVLat PN photostimulation, with a range of direct and indirect connections identified and evidence of a postsynaptic population of inhibitory interneurons. Together, these findings confirm a substantial role for PNs in local spinal sensory processing, as well as relay of sensory signals to the brain.

Abstract Determining the factors influencing habitat selection and hunting success in top predators is crucial for understanding how these species may respond to environmental changes. For marine top predators, such factors have been documented in pelagic foragers, with habitat use and hunting success being linked to chlorophyll-a concentrations, sea surface temperature and light conditions. In contrast, little is known about the determinants of benthic marine predators. The Australian fur seal (Arctocephalus pusillus doriferus) is a benthic-diving forager that has a breeding and foraging distribution largely restricted to Bass Strait, the shallow (max. depth 80 m) continental shelf region between the Australian mainland and Tasmania. The species forages mostly on benthic prey and represents the greatest resident marine predator biomass in south-eastern Australia. The region is also one of the world’s fastest-warming marine areas and oceanographic changes are influencing shifts in prey distribution and abundance. In the present study, GPS-derived locations of benthic dives (n = 288,449) and dive behaviour metrics were used to determine seafloor habitat selection and factors influencing hunting success in 113 lactating adult females from Kanowna Island during the winters of 2006–2021. Individuals non-randomly selected foraging habitats comprised of deeper, steeper sloped, muddy-sandy areas with less gravel and highly disturbed regions (P 40 m) characterised by moderate presence of gravel (25–50%) and substantial rock composition (50–75%) on the seabed. These findings suggest that habitat use and hunting success in adult female Australian fur seals could be impacted by predicted oceanographic changes, such as rising temperature, altered currents and waves which may modify seafloor characteristics and benthic communities.

Abstract Background A significant issue with innovative problem-solving in healthcare is an existing deficiency in continuing education for many healthcare professionals, which hinders the successful implementation of inventive solutions and progress in the field. Educators play a crucial role in guiding students to cultivate the knowledge and skills necessary to confront these challenges, including problem solving, collaboration, and the use of rapidly advancing technologies. It is vital to design educational programs that empower and motivate students to develop the proficiency and knowledge they need to be effective problem solvers, collaborators, and cultivators of innovative solutions. This project aims to assess the implementation and effectiveness of a codesigned postgraduate university program for a multidisciplinary health workforce. Methods The Leading Health Services Innovation Project is a hybrid type 2 mixed method implementation trial of a codesigned Graduate Certificate in Health Services Innovation. In collaboration with a large tertiary and quaternary health service, we developed a codesign process to guide the project, with time quarantined to create space for two-way learning between health sector partners and healthcare academics. Qualitative interviews and quantitative surveys for primary users will evaluate the implementation strategies. The reach, effectiveness, adoption implementation, and maintenance (RE-AIM) framework will guide the evaluation and maintenance of the program. Results Integrating a codesign strategy complemented by a well-structured implementation and evaluation protocol that is a combination of implementation science theoretical frameworks (Knowledge to Action, Evidence-Based Co-design, RE-AIM) may lead to translational competence as a potential outcome. Anticipated outcomes The application, resourcing and commitment to codesigned tertiary-level learning and qualification will demonstrate the achievement of a contemporary and comprehensive postgraduate university degree program in health innovation management.

Abstract The Clinical and Laboratory Standards Institute (CLSI) M27 guidelines are the recommended and most commonly used protocols for broth microdilution antifungal susceptibility testing of yeasts. However, these guidelines are limited to the use of 96-well assay plates, limiting assay capacity. With the increased risk of fungal resistance emerging in the community, it is important to have alternative protocols available, that offer higher throughput and can screen more than eight to ten potential antifungal compounds per plate. This study presents an optimised broth microdilution minimum inhibitory concentration (MIC) method for testing the susceptibility of yeasts in an efficient high throughput screening setup, with minimal growth variability and maximum reproducibility. We extend the M27 guidelines and optimise the conditions for 384-well plates. Validation of the assay was performed with ten clinically used antifungals (fluconazole, amphotericin B, 5-fluorocytosine, posaconazole, voriconazole, ketoconazole, itraconazole, caspofungin diacetate, anidulafungin and micafungin) against Candida albicans and Cryptococcus neoformans.

Abstract The roles of sex hormones such as estradiol, testosterone, and sex hormone-binding globulin (SHBG) in the etiology of lung and colorectal cancers in women, among the most common cancers after breast cancer, are unclear. This Mendelian randomization (MR) study evaluated such potential causal associations in women of European ancestry. We used summary statistics data from genome-wide association studies on sex hormones and from the Trøndelag Health Study (HUNT) and large consortia on cancers. There was suggestive evidence of 1-standard deviation increase in total testosterone levels being associated with a lower risk of lung non-adenocarcinoma (hazard ratio 0.60, 95% confidence interval 0.37–0.98) in the HUNT Study. However, this was not confirmed by using data from a larger consortium. In general, we did not find convincing evidence to support a causal role of sex hormones on risk of lung and colorectal cancers in women of European ancestry.

Abstract We present the theory, concept and design of an efficient, megawatt coherent Cherenkov radiation source based on a two-dimensional periodic surface lattice (2D-PSL) cavity combined with a novel energy recovery system for the generation of highly efficient (> 50%) single-frequency radiation. We demonstrate the scalability of the transverse dimension of the 2D-PSL cavity of the Cherenkov source and thus the potential for efficient, continuous-wave, high-power (> 1 MW) operation; fundamental to the eventual realization of clean, fusion energy. These new sources, with the capacity to operate in the 0.1-10THz range, hold strong promise to address the long-standing “Terahertz gap”. By combining a Cherenkov oscillator driven by a non-gyrating beam with an innovative four-stage depressed collector energy recovery system, the overall device efficiency can be increased to be competitive with gyrotrons in the requirements for heating and current drive in fusion plasma. In these Cherenkov devices, the frequency independence of the magnetic guide field enables advantageous frequency scaling without deployment constraints, making them especially attractive for high-impact applications in fusion science, turbulence diagnostics, non-destructive testing and biochemical spectroscopy. The novel energy recovery techniques presented in this paper have broad applicability to many electron-beam driven devices, bringing revolutionary potential to future THz source technologies.

Abstract The glycoside hydrolase family 3 (GH3) β-glucosidases from filamentous fungi are crucial industrial enzymes facilitating the complete degradation of lignocellulose, by converting cello-oligosaccharides and cellobiose into glucose. Understanding the diverse domain organization is essential for elucidating their biological roles and potential biotechnological applications. This research delves into the variability of domain organization within GH3 β-glucosidases. Two distinct configurations were identified in fungal GH3 β-glucosidases, one comprising solely the GH3 catalytic domain, and another incorporating the GH3 domain with a C-terminal fibronectin type III (Fn3) domain. Notably, Streptomyces filamentous bacteria showcased a separate clade of GH3 proteins linking the GH3 domain to a carbohydrate binding module from family 2 (CBM2). As a first step to be able to explore the role of accessory domains in β-glucosidase activity, a screening system utilizing the well-characterised Aspergillus niger β-glucosidase gene (bglA) in bglA deletion mutant host was developed. Based on this screening system, reintroducing the native GH3-Fn3 gene successfully expressed the gene allowing detection of the protein using different enzymatic assays. Further investigation into the role of the accessory domains in GH3 family proteins, including those from Streptomyces, will be required to design improved chimeric β-glucosidases enzymes for industrial application.

Abstract The objective of this work is to showcase the ortho-positronium lifetime as a probe for soft-tissue characterization. We employed positron annihilation lifetime spectroscopy to experimentally measure the three components of the positron annihilation lifetime—para-positronium (p-Ps), positron, and ortho-positronium (o-Ps)—for three types of porcine, non-fixated soft tissues ex vivo: adipose, hepatic, and muscle. Then, we benchmarked our measurements with X-ray phase-contrast imaging, which is the current state-of-the-art for soft-tissue analysis. We found that the o-Ps lifetime in adipose tissues (2.54 ± 0.12 ns) was approximately 20% longer than in hepatic (2.04 ± 0.09 ns) and muscle (2.03 ± 0.12 ns) tissues. In addition, the separation between the measurements for adipose tissue and the other tissues was better from o-Ps lifetime measurement than from X-ray phase-contrast imaging. This experimental study proved that the o-Ps lifetime is a viable non-invasive probe for characterizing and classifying the different soft tissues. Specifically, o-Ps lifetime as a soft-tissue characterization probe had a strong sensitivity to the lipid content that can be potentially implemented in commercial positron emission tomography scanners that feature list-mode data acquisition.

Abstract PDZ domain mediated interactions with voltage-gated calcium (CaV) channel C-termini play important roles in localizing membrane Ca2+ signaling. The first such interaction was described between the scaffolding protein Mint-1 and CaV2.2 in mammals. In this study, we show through various in silico analyses that Mint is an animal-specific gene with a highly divergent N-terminus but a strongly conserved C-terminus comprised of a phosphotyrosine binding domain, two tandem PDZ domains (PDZ-1 and PDZ-2), and a C-terminal auto-inhibitory element that binds and inhibits PDZ-1. In addition to CaV2 chanels, most genes that interact with Mint are also deeply conserved including amyloid precursor proteins, presenilins, neurexin, and CASK and Veli which form a tripartite complex with Mint in bilaterians. Through yeast and bacterial 2-hybrid experiments, we show that Mint and CaV2 channels from cnidarians and placozoans interact in vitro, and in situ hybridization revealed co-expression in dissociated neurons from the cnidarian Nematostella vectensis. Unexpectedly, the Mint orthologue from the ctenophore Hormiphora californiensis strongly bound the divergent C-terminal ligands of cnidarian and placozoan CaV2 channels, despite neither the ctenophore Mint, nor the placozoan and cnidarian orthologues, binding the ctenophore CaV2 channel C-terminus. Altogether, our analyses suggest that the capacity of Mint to bind CaV2 channels predates bilaterian animals, and that evolutionary changes in CaV2 channel C-terminal sequences resulted in altered binding modalities with Mint.

BackgroundVenous thromboembolism (VTE) and atrial fibrillation (AF) disproportionately affect older adults, who are at increased risk of bleeding from treatment with anticoagulant therapy. The impact of bleeding on older adults' quality of life (QoL) is poorly understood due to the lack of a validated measure of their experience. This study's purpose is to describe the first evidence-based steps in developing a new condition-specific patient-reported outcome measure (PROM) for the effect of anticoagulant-related bleeding on older adults' QoL.MethodsAdults aged 65 years and older with VTE or AF at the University of Utah who were eligible for anticoagulation were recruited. We purposely sampled by age, sex, race/ethnicity, diagnosis, bleeding history and anticoagulant. In accordance with evidence-based guidelines for PROM development, focus groups were conducted to refine a hypothesized conceptual model of bleeding-related QoL and analyzed according to hybrid inductive and deductive thematic analysis methods. Focus group results were used to write a draft PROM. Semi-structured qualitative interviews were conducted to revise items on the draft PROM and were analyzed using thematic analysis to generate a final bleeding-related QoL PROM.ResultsTwenty individuals participated in interviews, and nine in two focus groups. Median age was 81 (range 69-95), 52% were women and 69% were White (10% Asian, 10% Hispanic, 7% Black, 3% Native Hawaiian/Pacific Islander). Through inductive and deductive thematic analysis of focus groups, we identified 19 themes (e.g., specific bleeding symptoms, adaptations in relationships, fatigue) that fell into 5 domains: 1) bleeding symptoms, 2) healthcare experiences, 3) social function, 4) emotional function, 5) physical function. These 19 themes and 5 domains formed the basis of the draft PROM. We conducted semi-structured cognitive interviews, and we performed thematic analysis to revise the drafted measure for clarity and relevance. The data resulted in a new PROM for bleeding-related QoL in older adults.ConclusionEmploying evidence-based methods for PROM development, we found that bleeding can have a marked impact on everyday activities, emotional wellbeing and interpersonal relations for older adults. We incorporated these distinct aspects of wellbeing and function into a novel PROM for bleeding-related QoL that can inform clinical care and research after subsequent psychometric validation.

Sedentism is an adaptive alternative in human societies which is often associated with the emergence of complex societies in the Holocene. To elucidate the factors and processes of the emergence of sedentary societies, continuous accumulation of case studies based on robust evidence from across the world is required. Given abundant archaeological and geological evidence from the late Pleistocene to early Holocene, Tanegashima Island, situated in the southern Japanese Archipelago of the northwestern Pacific Rim, has significant potential to unravel factors and processes of sedentism. Our study evaluates long-term change in hunter-gatherer mobility on Tanegashima Island from the Upper Paleolithic to Incipient Jomon (ca.36,000-12,800 cal BP). Based on Bayesian age modelling, we performed diachronic analyses on lithic toolkit structure, lithic reduction technology, lithic raw material composition, and occupation intensity. The results illustrate that settlement-subsistence strategies on Tanegashima primarily correspond to the change in environmental conditions, mainly food resources, and foragers increased their degrees of sedentism when abundant forest existed. More important is that highly stable sedentism, which is not observed until the Incipient Jomon, depends not only on such a productive environment, but also on the increase in population size. High occupation intensity during the Incipient Jomon on the island is likely attributed to an influx of people from Kyushu proper. Although the relationship between cause and effect of these factors is still to be clarified in future work, our study provides insights on the fundamental causes of sedentism in the temperate forest of the late Pleistocene.

Background Four Seriola species support recreational and commercial fisheries along the U.S. Atlantic Ocean and the Gulf of Mexico, with the S. dumerili Gulf of Mexico stock being overfished for over three decades. The study presented here is part of a fisheries-independent project initiated to determine an absolute abundance of S. dumerili, to expand biological knowledge of the species and to develop novel tools for fisheries management. Environmental DNA (eDNA) tools aimed at the detection and quantification of target species are starting to emerge in support of marine fisheries surveys. Key to progressing the field is Droplet Digital™ PCR (ddPCR™), a highly sensitive technique with advanced multiplexing and direct quantification capabilities that can provide fisheries scientists with improved interpretation of eDNA data. Methods We developed and validated a novel tetraplex ddPCR™ assay able to detect and distinguish between S. dumerili, S. fasciata, S. rivoliana, and S. zonata from seawater eDNA samples. In order to groundtruth ddPCR™ data, and explore its capacity to provide abundance estimates, we compared ddPCR™ detections and quantifications to abundance data inferred from multiple camera (ROV, S-BRUV, chevron trap) and acoustic (VPS array) gears deployed during a fisheries research gear-calibration cruise. Results We demonstrated that with eDNA contamination controls and best practice protocols, it is viable to conduct eDNA research as part of a fisheries survey cruise. eDNA sampling was completed in less time than camera gears (15 min vs 2 h). Both eDNA and camera gears detected the presence of S. dumerili and S. rivoliana at both sites and all sampling days, but not S. fasciata and S. zonata. eDNA concentration data was higher for S. dumerili than S. rivoliana at both sites for all sampling days, in line with abundance patterns obtained from camera gears. The highest correlation (r = 0.97) was obtained between the measures of eDNA between gear deployments and ROV. Discussion Incorporating eDNA in fisheries surveys would not require additional days at sea and could improve precision in fish detection and abundance. eDNA can be a valuable complement to camera gears deployed in geographic areas or seasons with poor visibility conditions, where fish may be present but cannot be confidently identified to the species level. The high correlation obtained between ROV and eDNA data collected between gear deployments adds to a growing number of studies demonstrating the potential of eDNA as an indicator of abundance for fisheries stock assessments. Time-series data from a carefully designed eDNA survey, that estimates relative abundance, could be used as an index of relative abundance for the S. dumerili stock assessment. To achieve this, investment into follow-up studies with increased sample sizes and spatial and temporal replication would be necessary to allow for year-to-year comparisons and validate the robustness of the correlation observed.

Summary: Background: Perivascular spaces (PVS) on brain MRI are surrogates for small parenchymal blood vessels and their perivascular compartment, and may relate to brain health. However, it is unknown whether PVS can predict dementia risk and brain atrophy trajectories in participants without dementia, as longitudinal studies on PVS are scarce and current methods for PVS assessment lack robustness and inter-scanner reproducibility. Methods: We developed a robust algorithm to automatically assess PVS count and size on clinical MRI, and investigated 1) their relationship with dementia risk and brain atrophy in participants without dementia, 2) their longitudinal evolution, and 3) their potential use as a screening tool in simulated clinical trials. We analysed 46,478 clinical measurements of cognitive functioning and 20,845 brain MRI scans from 10,004 participants (71.1 ± 9.7 years-old, 56.6% women) from three publicly available observational studies on ageing and dementia (the Alzheimer’s Disease Neuroimaging Initiative, the National Alzheimer’s Coordinating Centre database, and the Open Access Series of Imaging Studies). Clinical and MRI data collected between 2004 and 2022 were analysed with consistent methods, controlling for confounding factors, and combined using mixed-effects models. Findings: Our fully-automated method for PVS assessment showed excellent inter-scanner reproducibility (intraclass correlation coefficients >0.8). Fewer PVS and larger PVS diameter at baseline predicted higher dementia risk and accelerated brain atrophy. Longitudinal trajectories of PVS markers differed significantly in participants without dementia who converted to dementia compared with non-converters. In simulated placebo-controlled trials for treatments targeting cognitive decline, screening out participants at low risk of dementia based on our PVS markers enhanced the power of the trial independently of Alzheimer’s disease biomarkers. Interpretation: These robust cerebrovascular markers predict dementia risk and brain atrophy and may improve risk-stratification of patients, potentially reducing cost and increasing throughput of clinical trials to combat dementia. Funding: US National Institutes of Health.

Background Gait analysis is traditionally conducted using marker-based methods yet markerless motion capture is emerging as an alternative. Initial studies have begun to evaluate the reliability of markerless motion capture yet the evaluation of different clothing conditions across sessions and complete evaluation of the lower limb and pelvis reliability have yet to be considered. The aim of this study was to evaluate the inter-trial, inter-session and inter-session-clothing variation and root mean square differences between tight- or loose-fitting clothing during walking. Method Twenty-two healthy adult participants walked along an indoor walkway whilst eight video cameras recorded their gait in either tight- or loose-fitting clothing. A commercial markerless motion capture system (Theia3D) provided gait kinematics for evaluation. Results Reliability results showed average inter-trial variation of

Background Although rare, uterine sarcomas account for a high proportion of uterine cancer mortality. Treatment options and robust trial data are limited.Objectives The TOURISM study (Treatment Outcomes in UteRIne SarcoMa) is a UK-wide study by the National Oncology Trainees Collaborative for Healthcare Research which aimed to characterise this patient cohort.Design A retrospective descriptive cohort study. Patients with carcinosarcomas/mixed Mullerian tumours, non-uterine gynaecological sarcomas and uterine metastases were excluded. Routine clinical data, including general patient demographics, diagnosis, treatment and outcomes, were collated and pseudonymised.Setting Patients diagnosed with uterine sarcoma in the UK National Health Service between 1 January 2008 and 31 December 2017 were identified from electronic records.Participants A total of 406 patients from eight centres were eligible for inclusion.Results The median age at diagnosis was 56 years, with leiomyosarcoma the most common diagnosis (54.4%). The majority (57.9%) were diagnosed at the International Federation of Gynecology and Obstetrics stage I, with 19.7% diagnosed at stage IV. Nearly half (45.2%) of the patients received at least one line of chemotherapy, of which most (81.0%) received doxorubicin first-line. In the stage I group 7.4% received adjuvant chemotherapy and 15.0% received adjuvant radiotherapy. Median overall survival was 37 months; however, survival varied significantly by stage at diagnosis (stage I: 105 months; stage II: 33 months; stage III: 19 months; stage IV: 14 months).Conclusions Our data highlight the diversity in patient management in uterine sarcoma and a marked survival advantage for patients diagnosed with stage I disease. These data highlight the importance of a multidisciplinary approach and describe real-world trends in systemic therapies, radiotherapy and surgical treatment in this rare cancer type.