The measurement of young children's nocturnal sleep health and the development of the Perception of Infant and Toddler Sleep Scale (PoITSS) in Aotearoa New Zealand whanau (families).

OBJECTIVES: To modify an existing questionnaire Brief Infant Sleep Questionnaire - Revised (BISQ-R) to ensure that it is suitable to measure nocturnal sleep health in a diverse sample of young children from Aotearoa New Zealand whanau (families), and to develop a "Perception of Infant and Toddler Sleep Scale" (PoITSS) to use as a primary outcome measurement in an upcoming trial. METHODS: Items from the BISQ-R were adapted for use among ethnically diverse whanau, and tested online with caregivers of 0-2 year old children. A PoITSS score was generated by scaling the responses from three of the questionnaire items to create a value between 0 (very poor) and 10 (very good). Caregivers provided qualitative feedback about the ease of interpreting and answering questionnaire items. RESULTS: Caregivers of 957 children (35% Maori, 12% Pacific) completed the questionnaire. Few differences in children's nocturnal sleep were observed by demographic characteristics. The mean PoITSS score was 6.9 (SD 2.3) and was slightly higher among Maori children (mean difference 0.4, 95% CI 0.1, 0.7). Test-retest indicated good reliability (ICC=0.81). While the majority (86%) of caregivers did not find it difficult to answer any of the items which formed the PoITSS, qualitative feedback indicated that simple modifications to some items would help ensure that they would be well understood by most caregivers. CONCLUSIONS: Items from the BISQ-R were successfully adapted, and the PoITSS scale was shown to be appropriate, for use in ethnically diverse Aotearoa New Zealand whanau with young children.

An atlas of GPCRs in dopamine neurons: Identification of the free fatty acid receptor 4 as a regulator of food and water intake.

Midbrain dopaminergic neurons (DANs) are subject to extensive metabotropic regulation, but the repertoire of G protein-coupled receptors (GPCRs) present in these neurons has not been mapped. Here, we isolate DANs from Dat-eGFP mice to generate a GPCR atlas by unbiased qPCR array expression analysis of 377 GPCRs. Combined with data mining of scRNA-seq databases, we identify multiple receptors in DAN subpopulations with 38 of these receptors representing the majority of transcripts. We identify 41 receptors expressed in midbrain DANs but not in non-DAN midbrain cells, including the free fatty acid receptor 4 (FFAR4). Functional expression of FFAR4 is validated by ex vivo Ca2+ imaging, and in vivo experiments support that FFAR4 negatively regulates food and water intake and bodyweight. In addition to providing a critical framework for understanding metabotropic DAN regulation, our data suggest fatty acid sensing by FFAR4 as a mechanism linking high-energy intake to the dopamine-reward pathway.

Effects of Different Protein Sources on Amino Acid Absorption and Plasma Appearance of Tryptophan, Large Neutral Amino Acids, and Tryptophan Metabolites in Pigs.

BACKGROUND: Absorption of tryptophan (TRP) across the gut epithelium is potentially modulated by competing large neutral amino acids (LNAAs), which could affect the appearance of TRP and its metabolites in the bloodstream. OBJECTIVES: This study aimed to determine, in a growing pig model of an adult human, the absorption of TRP and other LNAAs from the gastrointestinal tract, and plasma appearance of TRP, LNAAs, and TRP metabolites, in response to dietary proteins varying in TRP content. METHODS: Pigs were adapted for 7 d to each of 4 diets that differed in their protein source and TRP content: 1) alpha-lactalbumin (AL; 9.95 mg TRP/g diet DM), 2) whey protein (6.59 mg TRP/g), 3) casein (3.73 mg TRP/g), or 4) zein (0.14 mg TRP/g). On day 8, pigs were euthanised after a 12-h fast (baseline), or 1, 2, 3, 4, or 6 h after they received a test meal consisting of 45 g protein, or a protein-free meal (n = 6 pigs at each time in each meal group). Tryptophan and LNAA absorption from the small intestine, and appearance of TRP, LNAAs, and TRP metabolites (melatonin, serotonin, kynurenine pathway metabolites), in the portal vein and systemic circulation, were determined. RESULTS: AL intake resulted in sustained elevated plasma TRP concentrations after an overnight fast. The amount of TRP absorbed was dose-dependently related to protein TRP content (P = 0.028), with fastest rates for pigs fed AL (371 mg/h). Portal and systemic plasma TRP, TRP/LNAA, and the TRP metabolites were highest (P ≤ 0.05) after AL intake, and remained above baseline levels for ∼4 h postprandially. Absorption rates of TRP correlated with postprandial plasma TRP and TRP metabolites (P ≤ 0.05). CONCLUSIONS: In adult humans, postprandial plasma TRP and TRP metabolite concentrations can likely be modulated by the TRP content of the meal.

Creating a health informatics data resource for hearing health research.

BACKGROUND: The National Institute of Health and Social Care Research (NIHR) Health Informatics Collaborative (HIC) for Hearing Health has been established in the UK to curate routinely collected hearing health data to address research questions. This study defines priority research areas, outlines its aims, governance structure and demonstrates how hearing health data have been integrated into a common data model using pure tone audiometry (PTA) as a case study. METHODS: After identifying key research aims in hearing health, the governance structure for the NIHR HIC for Hearing Health is described. The Observational Medical Outcomes Partnership (OMOP) was chosen as our common data model to provide a case study example. RESULTS: The NIHR HIC Hearing Health theme have developed a data architecture outlying the flow of data from all of the various siloed electronic patient record systems to allow the effective linkage of data from electronic patient record systems to research systems. Using PTAs as an example, OMOPification of hearing health data successfully collated a rich breadth of datapoints across multiple centres. CONCLUSION: This study identified priority research areas where routinely collected hearing health data could be useful. It demonstrates integration and standardisation of such data into a common data model from multiple centres. By describing the process of data sharing across the HIC, we hope to invite more centres to contribute and utilise data to address research questions in hearing health. This national initiative has the power to transform UK hearing research and hearing care using routinely collected clinical data.

Universal irradiation of platelets: Does irradiation affect the quality, effectiveness, and safety of platelets for transfusion?

We aimed to identify any detrimental effects on platelet quality and clinical effectiveness, of irradiated platelets compared to non-irradiated platelets for transfusion. The review was conducted in accordance with PRISMA guidelines. The protocol was prospectively registered on PROSPERO [CRD42023441930]. Our search identified 3002 references, of which we included 44 studies. Forty-one were in vitro only studies, two studies were in healthy volunteers, and one study reported clinical outcomes in thrombocytopenic patients. X-ray was used exclusively in three studies, and alongside gamma irradiation in one study. Two studies did not report the source of irradiation. The remaining 38 studies used gamma irradiation only. We assessed risk of bias (ROB) for studies reporting clinical and in vivo outcomes using ROB 2.0 (3 studies). We adapted a ROB tool designed for animal studies to assess ROB for the studies reporting in vitro outcomes (43 studies). We assessed the certainty of the evidence for the eight outcomes deemed most important to assess platelet quality and clinical effectiveness (where day 0 is the day of the blood draw). Overall, gamma irradiation has little to no effect on most markers of platelet quality and effectiveness. Where there is evidence of detriment from irradiation, differences are small in vitro, and are unlikely to affect clinical outcomes following transfusion. However, the evidence base is limited. Only half the studies could be included in any analysis. There is very limited evidence for x-ray as a source of irradiation and, given the potential benefits of using x-ray over gamma irradiation (ease of use and safety requirements), we would welcome further research comparing x-ray to gamma, and x-ray to a non-irradiated control.

Global terrestrial drought and its projected socioeconomic implications under different warming targets.

Droughts are increasingly frequent as the Earth warms, presenting adaptation challenges for ecosystems and human communities worldwide. A strategic environmental assessment (SEA) and the integration of adaptation strategies into policies, plans, and programs (PPP) are two important approaches for enhancing climate resilience and fostering sustainable development. This study developed an innovative approach to strengthen the SEA of droughts by quantifying the impacts of future temperature increases. A novel method for projecting drought events was integrated into the SEA process by leveraging multiple data sources, including atmospheric reanalysis, reconstructions, satellite-based observations, and model simulations. We identified drought conditions using terrestrial water storage (TWS) anomalies and applied a random forest (RF) model for disentangling the drivers behind drought events. We then set two global warming targets (2.0 °C and 2.5 °C) and analyzed drought changes under three shared socioeconomic pathways (SSP126, SSP370, SSP585). In a 2.0 °C warming world, over 50 % of the global surface will face increased drought risk. With an additional 0.5 °C increase, >60 % of the land will be prone to further drought escalation. We utilized copulas to build the joint distribution for drought duration and severity, estimating the joint return periods (JRP) for bivariate drought hazard. In tropical and subtropical regions, JRP reductions exceeding half are projected for >33 % of the regional land surface under 2.0 °C warming and for >50 % under 2.5 °C warming. Finally, we projected the impacts of drought events on population and gross domestic product (GDP). Among the three SSPs, under SSP370, population exposure is highest and GDP exposure is minimal under 2.0 °C warming. Global GDP and population risks from drought are projected to increase by 37 % and 24 %, respectively, as warming continues. This study enhances the accuracy of SEA in addressing drought risks and vulnerabilities, supporting climate-resilient planning and adaptive strategies.

Process-Informed Subsampling Improves Subseasonal Rainfall Forecasts in Central America.

Subseasonal rainfall forecast skill is critical to support preparedness for hydrometeorological extremes. We assess how a process-informed evaluation, which subsamples forecasting model members based on their ability to represent potential predictors of rainfall, can improve monthly rainfall forecasts within Central America in the following month, using Costa Rica and Guatemala as test cases. We generate a constrained ensemble mean by subsampling 130 members from five dynamic forecasting models in the C3S multimodel ensemble based on their representation of both (a) zonal wind direction and (b) Pacific and Atlantic sea surface temperatures (SSTs), at the time of initialization. Our results show in multiple months and locations increased mean squared error skill by 0.4 and improved detection rates of rainfall extremes. This method is transferrable to other regions driven by slowly-changing processes. Process-informed subsampling is successful because it identifies members that fail to represent the entire rainfall distribution when wind/SST error increases.

mRNA Display in Cell Lysates Enables Identification of Cyclic Peptides Targeting the BRD3 Extraterminal Domain.

mRNA display is a powerful technology to screen libraries of >1012 cyclic peptides against a protein target, enabling the rapid discovery of high affinity ligands. These cyclic peptides are particularly well suited to challenging protein targets that have been difficult to drug with small molecules. However, target choice can still be limited as screens are typically performed against purified proteins which often demands the use of isolated domains and precludes the use of aggregation-prone targets. Herein, we report a method to perform mRNA display selections in mammalian cell lysates without the need for prior target purification, vastly expanding the potential target scope of mRNA display. We have applied the methodology to identify low to sub-nanomolar peptide binders for two targets: a NanoLuc subunit (LgBiT) and full-length bromodomain-containing protein 3 (BRD3). Our cyclic peptides for BRD3 were found to bind to the extraterminal (ET) domain of BRD3 and the closely related BRD proteins, BRD2 and BRD4. While many chemical probes exist for the bromodomains of BRD proteins, the ET domain is relatively underexplored, making these peptides valuable additions to the BRD toolbox.

Extension of the circulatory half-life of recombinant ecallantide via albumin fusion without loss of anti-kallikrein activity.

Ecallantide comprises Kunitz Domain 1 of Tissue Factor Pathway Inhibitor, mutated at seven amino acid positions to inhibit plasma kallikrein (PK). It is used to treat acute hereditary angioedema (HAE). We appended hexahistidine tags to the N- or C-terminus of recombinant Ecallantide (rEcall) and expressed and purified the resulting proteins, with or without fusion to human serum albumin (HSA), using Pichia pastoris. The inhibitory constant (Ki) of rEcall-H6 or H6-rEcall for PK was not increased by albumin fusion. When 125I-labelled rEcall proteins were injected intravenously into mice, the area under the clearance curve (AUC) was significantly increased, 3.4- and 3.6-fold, for fusion proteins H6-rEcall-HSA and HSA-rEcall-H6 versus their unfused counterparts but remained 2- to 3-fold less than that of HSA-H6. The terminal half-life of H6-rEcall-HSA and HSA-H6 did not differ, although that of HSA-rEcall-H6 was significantly shorter than either other protein. Receptor Associated Protein (RAP), a Low-density lipoprotein Receptor-related Protein (LRP1) antagonist, competed H6-rEcall-HSA clearance more effectively than intravenous immunoglobulin (IVIg), a neonatal Fc receptor (FcRn) antagonist. HSA fusion decreases rEcall clearance in vivo, but LRP1-mediated clearance remains more important than FcRn-mediated recycling for rEcall fusion proteins. The properties of H6-rEcall-HSA warrant investigation in a murine model of HAE.

Macroplastic Fate and Transport Modeling: Freshwaters Act as Main Reservoirs.

Macroplastic fate and transport in the freshwater environment are of great concern due to the potentially harmful effects of macroplastic on plants, animals, and humans. Here, we present a modeling approach to simulate macroplastic fate and transport at the country scale based on an existing plastic release model. The fate model was parametrized through available monitoring data and results from field experiments and applied to Swiss rivers and lakes. We found that almost all (98%) macroplastic emissions into freshwater remain within Switzerland. After exploring the influences of weirs, retention in rivers, and retention in lakes through a sensitivity analysis, we found a high retention variability across different catchments and within rivers. In all 22 analyzed scenarios for continuous retention along each river bank (i.e., beaching), we found that at least 70% of input emissions into the water bodies would be retained long-term in the catchments (about 200 g per river km and year). Across all catchments, we found a dominance of "continuous retention" through beaching along the entire river length compared with "point retention" at weirs or lakes. Thus, by modeling macroplastic fate and transport on a country level for the first time, we were able to confirm the concept of "rivers as plastic reservoirs" through modeling.

Investigating the potential of X chromosome shredding for mouse genetic biocontrol.

CRISPR-Cas9 technology has facilitated development of strategies that can potentially provide more humane and effective methods to control invasive vertebrate species, such as mice. One promising strategy is X chromosome shredding which aims to bias offspring towards males, resulting in a gradual and unsustainable decline of females. This method has been explored in insects with encouraging results. Here, we investigated this strategy in Mus musculus by targeting repeat DNA sequences on the X chromosome with the aim of inducing sufficient DNA damage to specifically eliminate X chromosome-bearing sperm during gametogenesis. We tested three different guide RNAs (gRNAs) targeting different repeats on the X chromosome, together with three male germline-specific promoters for inducing Cas9 expression at different stages of spermatogenesis. A modest bias towards mature Y-bearing sperm was detected in some transgenic males, although this did not translate into significant male-biasing of offspring. Instead, cleavage of the X chromosome during meiosis typically resulted in a spermatogenic block, manifest as small testes volume, empty tubules, low sperm concentration, and sub/infertility. Our study highlights the importance of controlling the timing of CRISPR-Cas9 activity during mammalian spermatogenesis and the sensitivity of spermatocytes to X chromosome disruption.

Supporting healthy lifestyles for First Nations women and communities through co-design: lessons and early findings from remote Northern Australia.

Background: The period before, during, and after pregnancy presents an opportunity to reduce diabetes-related risks, which in Australia disproportionately impact Aboriginal and Torres Strait Islander women. Collaboration with Aboriginal and Torres Strait Islander women/communities is essential to ensure acceptability and sustainability of lifestyle modifications. Using a novel co-design approach, we aimed to identify shared priorities and potential lifestyle strategies. We also reflected on learnings from this approach. Methods: We conducted 11 workshops and 8 interviews at two sites in Australia's Northern Territory (Central Australia and Top End), using experience-based co-design (EBCD) and incorporating principles of First Nations participatory research. Workshops/interviews explored participant' experiences and understanding of diabetes in pregnancy, contextual issues, and potential lifestyle strategies. Participants included three groups: 1) Aboriginal and Torres Strait Islander women of reproductive age (defined as aged 16-45 years); 2) Aboriginal and Torres Strait Islander community members; and 3) health/community services professionals. The study methodology sought to amplify the voices of Aboriginal women. Findings: Participants included 23 women between ages 16-45 years (9 with known lived experience of diabetes in pregnancy), 5 community members and 23 health professionals. Key findings related to identified priority issues, strategies to address priorities, and reflections on use of EBCD methodology. Priorities were largely consistent across study regions: access to healthy foods and physical activity; connection to traditional practices and culture; communication regarding diabetes and related risks; and the difficulty for women of prioritising their health among competing priorities. Strategies included implementation of a holistic women's program in Central Australia, while Top End participants expressed the desire to improve nutrition, peer support and community awareness of diabetes. EBCD provided a useful structure to explore participants' experiences and collectively determine priorities, while allowing for modifications to ensure co-design methods were contextually appropriate. Challenges included the resource-intensive nature of stakeholder engagement, and collaborating effectively with services and communities when researchers were "outsiders". Conclusions: A hybrid methodology using EBCD and First Nations participatory research principles enabled collaboration between Aboriginal women, communities and health services to identify shared priorities and solutions to reduce diabetes-related health risks. Genuine co-design processes support self-determination and enhance acceptability and sustainability of health strategies.

River plastic transport and storage budget.

Rivers are one of the main conduits that deliver plastic from land into the sea, and also act as reservoirs for plastic retention. Yet, our understanding of the extent of river exposure to plastic pollution remains limited. In particular, there has been no comprehensive quantification of the contributions from different river compartments, such as the water surface, water column, riverbank and floodplain to the overall river plastic transport and storage. This study aims to provide an initial quantification of these contributions. We first identified the main relevant transport processes for each river compartment considered. We then estimated the transport and storage terms, by harmonizing available observations on surface, suspended and floodplain plastic. We applied our approach to two river sections in The Netherlands, with a focus on macroplastics (≥2.5 cm). Our analysis revealed that for the studied river sections, suspended plastics account for over 96% of item transport within the river channel, while their relative contribution to mass transport is only 30%-37% (depending on the river section considered). Surface plastics predominantly consisted of heavier items (mean mass: 7.1 g/#), whereas suspended plastics were dominated by lighter fragments (mean mass: 0.1 g/#). Additionally, the majority (98%) of plastic mass was stored within the floodplains, with the river channel accounting for only 2% of the total storage. Our study developed a harmonized approach for quantifying plastic transport and storage across different river compartments, providing a replicable methodology applicable to different regions. Our findings emphasize the importance of systematic monitoring programs across river compartments for comprehensive insights into riverine plastic pollution.

Water hyacinths retain river plastics.

Rivers represent one of the main conduits for the delivery of plastics to the sea, while also functioning as reservoirs for plastic retention. In tropical regions, rivers are exposed to both high levels of plastic pollution and invasion of water hyacinths. This aquatic plant forms dense patches at the river surface that drift due to winds and currents. Recent work suggests that water hyacinths play a crucial role in influencing plastic transport, by efficiently trapping the majority of surface plastic within their patches. However, a comprehensive understanding of the interaction between water hyacinths and plastics is still lacking. We hypothesize that the properties relevant to plastic transport change due to their trapping in water hyacinth patches. In particular, the length scale, defined as the characteristic size of the transported material, is a key property in understanding how materials move within rivers. Here, we show that water hyacinth patches trap on average 54%-77% of all observed surface plastics at the measurement site (Saigon river, Vietnam). Both temporally and spatially, we found that plastic and water hyacinth presence co-occur. The formation of plastic-plant aggregates carries significant implications for both clean-up and monitoring purposes, as these aggregates can be detected from space and need to be jointly removed. In addition, the length scale of trapped plastics (∼4.0 m) was found to be forty times larger than that of open water plastics (∼0.1 m). The implications of this increased length scale for plastic transport dynamics are yet to be fully understood, calling for further investigation into travel distances and trajectories. The effects of plastic trapping likely extend to other key properties of plastic-plant aggregates, such as effective buoyancy and mass. Given the prevalence of plant invasion and plastic pollution in rivers worldwide, this research offers valuable insights into the complex environmental challenges faced by numerous rivers.

Emergence of fractal geometries in the evolution of a metabolic enzyme.

Fractals are patterns that are self-similar across multiple length-scales1. Macroscopic fractals are common in nature2-4; however, so far, molecular assembly into fractals is restricted to synthetic systems5-12. Here we report the discovery of a natural protein, citrate synthase from the cyanobacterium Synechococcus elongatus, which self-assembles into Sierpinski triangles. Using cryo-electron microscopy, we reveal how the fractal assembles from a hexameric building block. Although different stimuli modulate the formation of fractal complexes and these complexes can regulate the enzymatic activity of citrate synthase in vitro, the fractal may not serve a physiological function in vivo. We use ancestral sequence reconstruction to retrace how the citrate synthase fractal evolved from non-fractal precursors, and the results suggest it may have emerged as a harmless evolutionary accident. Our findings expand the space of possible protein complexes and demonstrate that intricate and regulatable assemblies can evolve in a single substitution.

Variations in proton transfer pathways and energetics on pristine and defect-rich quartz surfaces in water: Insights into the bimodal acidities of quartz.

HYPOTHESIS: Understanding the mechanisms of proton transfer on quartz surfaces in water is critical for a range of processes in geochemical, environmental, and materials sciences. The wide range of surface acidities (>9 pKa units) found on the ubiquitous mineral quartz is caused by the structural variations of surface silanol groups. Molecular scale simulations provide essential tools for elucidating the origin of site-specific surface acidities. SIMULATIONS: We used density-functional tight-binding-based molecular dynamics combined with rare-event metadynamics simulations to probe the mechanisms of deprotonation reactions from ten representative surface silanol groups found on both pristine and defect-rich quartz (101) surfaces with Si vacancies. FINDINGS: The results show that deprotonation is a highly dynamic process where both the surface hydroxyls and bridging oxygen atoms serve as the proton acceptors, in addition to water. Deprotonation of embedded silanols through intrasurface proton transfer exhibited lower pKa values with less H-bond participation and higher energy barriers, suggesting a new mechanism to explain the bimodal acidity observed on quartz surface. Defect sites, recently shown to comprise a significant portion of the quartz (101) surface, diversify the coordination and local H-bonding environments of the surface silanols, changing both the deprotonation pathways and energetics, leading to a wider range of pKa values (2.4 to 11.5) than that observed on pristine quartz surface (10.4 and 12.1).

Exhaled Volatile Organic Compounds for Asthma Control Classification in Children with Moderate to Severe Asthma: Results from the SysPharmPediA Study.

RATIONALE: Early identification of children with poorly controlled asthma is imperative for optimizing treatment strategies. The analysis of exhaled volatile organic compounds (VOCs) is an emerging approach to identify prognostic and diagnostic biomarkers in pediatric asthma. OBJECTIVES: To assess the accuracy of gas chromatography-mass spectrometry based exhaled metabolite analysis to differentiate between controlled and uncontrolled pediatric asthma. METHODS: This study encompassed a discovery (SysPharmPediA) and validation phase (U-BIOPRED, PANDA). Firstly, exhaled VOCs that discriminated asthma control levels were identified. Subsequently, outcomes were validated in two independent cohorts. Patients were classified as controlled or uncontrolled, based on asthma control test scores and number of severe attacks in the past year. Additionally, potential of VOCs in predicting two or more future severe asthma attacks in SysPharmPediA was evaluated. MEASUREMENTS AND MAIN RESULTS: Complete data were available for 196 children (SysPharmPediA=100, U-BIOPRED=49, PANDA=47). In SysPharmPediA, after randomly splitting the population into training (n=51) and test sets (n=49), three compounds (acetophenone, ethylbenzene, and styrene) distinguished between uncontrolled and controlled asthmatics. The area under the receiver operating characteristic curve (AUROCC) for training and test sets were respectively: 0.83 (95% CI: 0.65-1.00) and 0.77 (95% CI: 0.58-0.96). Combinations of these VOCs resulted in AUROCCs of 0.74 ±0.06 (UBIOPRED) and 0.68 ±0.05 (PANDA). Attacks prediction tests, resulted in AUROCCs of 0.71 (95% CI 0.51-0.91) and 0.71 (95% CI 0.52-0.90) for training and test sets. CONCLUSIONS: Exhaled metabolites analysis might enable asthma control classification in children. This should stimulate further development of exhaled metabolites-based point-of-care tests in asthma.