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We present a genome assembly from an individual male Platypus cylindrus (the oak pinhole borer; Arthropoda; Insecta; Coleoptera; Curculionidae). The genome sequence is 147.5 megabases in span. Most of the assembly is scaffolded into 8 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 19.29 kilobases in length. Gene annotation of this assembly on Ensembl identified 13,468 protein coding genes.

We present a genome assembly from an individual female Agrilus cyanescens (metallic wood-boring beetle; Arthropoda; Insecta; Coleoptera; Buprestidae). The genome sequence is 292.3 megabases in span. Most of the assembly is scaffolded into 10 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 15.91 kilobases in length.

We present a genome assembly from an individual female Entiminae weevil, Polydrusus pterygomalis (Arthropoda; Insecta; Coleoptera; Curculionidae). The genome sequence has a total length of 1,051.50 megabases. Most of the assembly is scaffolded into 11 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 20.95 kilobases in length.

We present a genome assembly from an individual Dogs-Mercury Flea Beetle, Hermaeophaga mercurialis (Arthropoda; Insecta; Coleoptera; Chrysomelidae). The genome sequence has a length of 479.40 megabases. Most of the assembly is scaffolded into 9 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 16.05 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,633 protein-coding genes.

We present a genome assembly from an individual female Clivina fossor (a ground beetle; Arthropoda; Insecta; Coleoptera; Carabidae). The genome sequence spans 612.60 megabases. Most of the assembly is scaffolded into 22 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 16.48 kilobases in length.

Abstract Sub-Saharan Africa is under-represented in global biodiversity datasets, particularly regarding the impact of land use on species’ population abundances. Drawing on recent advances in expert elicitation to ensure data consistency, 200 experts were convened using a modified-Delphi process to estimate ‘intactness scores’: the remaining proportion of an ‘intact’ reference population of a species group in a particular land use, on a scale from 0 (no remaining individuals) to 1 (same abundance as the reference) and, in rare cases, to 2 (populations that thrive in human-modified landscapes). The resulting bii4africa dataset contains intactness scores representing terrestrial vertebrates (tetrapods: ±5,400 amphibians, reptiles, birds, mammals) and vascular plants (±45,000 forbs, graminoids, trees, shrubs) in sub-Saharan Africa across the region’s major land uses (urban, cropland, rangeland, plantation, protected, etc.) and intensities (e.g., large-scale vs smallholder cropland). This dataset was co-produced as part of the Biodiversity Intactness Index for Africa Project. Additional uses include assessing ecosystem condition; rectifying geographic/taxonomic biases in global biodiversity indicators and maps; and informing the Red List of Ecosystems.

We present a genome assembly from an individual female Crepidodera aurea (flea beetle; Arthropoda; Insecta; Coleoptera; Chrysomelidae). The genome sequence is 509.0 megabases in span. Most of the assembly is scaffolded into 10 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 18.69 kilobases in length. Gene annotation of this assembly on Ensembl identified 19,944 protein coding genes.

We present a genome assembly from an individual male Platypus cylindrus (the oak pinhole borer; Arthropoda; Insecta; Coleoptera; Curculionidae). The genome sequence is 147.5 megabases in span. Most of the assembly is scaffolded into 8 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 19.29 kilobases in length. Gene annotation of this assembly on Ensembl identified 13,468 protein coding genes.

We present a genome assembly from an individual male Taphrorychus bicolor (the beech bark beetle; Arthropoda; Insecta; Coleoptera; Curculionidae). The genome sequence is 575.2 megabases in span. Most of the assembly is scaffolded into 12 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 15.46 kilobases in length. Gene annotation of this assembly on Ensembl identified 24,125 protein coding genes.

We present a genome assembly from an individual female Anaspis maculata (false flower beetle; Arthropoda; Insecta; Coleoptera; Scraptiidae). The genome sequence is 757.8 megabases in span. Most of the assembly is scaffolded into 8 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 16.31 kilobases in length. Gene annotation of this assembly on Ensembl identified 21,965 protein coding genes.

Abstract In many biomechanical analyses, the forces acting on a body during dynamic and static activities are often simplified as point loads. However, it is usually more accurate to characterize these forces as distributed loads, varying in magnitude and direction, over a given contact area. Evaluating these pressure distributions while they are applied to different parts of the body can provide effective insights for clinicians and researchers when studying health and disease conditions, for example when investigating the biomechanical factors that may lead to plantar ulceration in diabetic foot disease. At present, most processing and analysis for pressure data is performed using proprietary software, limiting reproducibility, transparency, and consistency across different studies. This paper describes an open-source software package, ‘pressuRe’, which is built in the freely available R statistical computing environment and is designed to process, analyze, and visualize pressure data collected on a range of different hardware systems in a standardized manner. We demonstrate the use of the package on pressure dataset from patients with diabetic foot disease, comparing pressure variables between those with longer and shorter durations of the disease. The results matched closely with those from commercially available software, and individuals with longer duration of diabetes were found to have higher forefoot pressures than those with shorter duration. By utilizing R’s powerful and openly available tools for statistical analysis and user customization, this package may be a useful tool for researchers and clinicians studying plantar pressures and other pressure sensor array based biomechanical measurements. With regular updates intended, this package allows for continued improvement and we welcome feedback and future contributions to extend its scope. In this article, we detail the package’s features and functionality.

Abstract Resistance mechanisms to immune checkpoint blockade therapy (ICBT) limit its response duration and magnitude. Paradoxically, Interferon γ (IFNγ), a key cytokine for cellular immunity, can promote ICBT resistance. Using syngeneic mouse tumour models, we confirm that chronic IFNγ exposure confers resistance to immunotherapy targeting PD-1 (α-PD-1) in immunocompetent female mice. We observe upregulation of poly-ADP ribosyl polymerase 14 (PARP14) in chronic IFNγ-treated cancer cell models, in patient melanoma with elevated IFNG expression, and in melanoma cell cultures from ICBT-progressing lesions characterised by elevated IFNγ signalling. Effector T cell infiltration is enhanced in tumours derived from cells pre-treated with IFNγ in immunocompetent female mice when PARP14 is pharmacologically inhibited or knocked down, while the presence of regulatory T cells is decreased, leading to restoration of α-PD-1 sensitivity. Finally, we determine that tumours which spontaneously relapse in immunocompetent female mice following α-PD-1 therapy upregulate IFNγ signalling and can also be re-sensitised upon receiving PARP14 inhibitor treatment, establishing PARP14 as an actionable target to reverse IFNγ-driven ICBT resistance.

We present a genome assembly from an individual female Malthinus flaveolus (soldier beetle; Arthropoda; Insecta; Coleoptera; Cantharidae). The genome sequence is 236.7 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 19.27 kilobases in length. Gene annotation of this assembly on Ensembl identified 16,617 protein coding genes.

We present a genome assembly from an individual male Lochmaea crataegi (the hawthorn leaf beetle; Arthropoda; Insecta; Coleoptera; Chrysomelidae). The genome sequence is 891.3 megabases in span. Most of the assembly is scaffolded into 16 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 18.32 kilobases in length.

We present a genome assembly from an individual female Agrilus cyanescens (metallic wood-boring beetle; Arthropoda; Insecta; Coleoptera; Buprestidae). The genome sequence is 292.3 megabases in span. Most of the assembly is scaffolded into 10 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 15.91 kilobases in length.

In real-world situations, marine fish farms accommodate multiple fish species and cohorts within the farm, leading to diverse farm layouts influenced by cage dimensions, configurations, and intricate arrangements. These cage management practices are essential to meet production demands, however, farm-level complexities can impact model predictions of waste deposition and benthic impact near fish cages. This is of particular importance when the cages are used for integrated multi-trophic aquaculture (IMTA) with benthic feeders, where this waste not only affects environmental conditions but also provides a potential food source. The Cage Aquaculture Particulate Output and Transport (CAPOT) model incorporated multiple species, cohorts, and cage arrangements to estimate waste distribution from a commercial fish farm in the Mediterranean between October 2018 and July 2019. This spreadsheet model estimated dispersion for individual fish cages using a grid resolution of 5 m x 5 m. The study categorized discrete production periods for each fish cage every month, aligning with intermittent changes in biomass and food inputs due to different cage management practices throughout production. This approach facilitated the use of detailed input data and enhanced model representativeness by considering variations in cage biomass, food types, settling velocities, and configurations. Model outputs, represented in contour plots, indicated higher deposition directly below fish cages that varied monthly throughout fish production cycles. Deposition footprints reflected changes in cage biomass, food inputs, and farm-level practices reflecting this real-world scenario where aquaculture does not follow a production continuum. Moreover, cohort dynamics and cage movements associated with the cage management practices of the fish farm influenced the quantity and fate of wastes distributed around fish cages, revealing variability in deposition footprints. Clearly, these findings have important implications for the design of benthic IMTA systems, with species such as sea cucumber and polychaetes. Variability in waste deposition creates challenges in identifying where the benthic organisms should be placed to allow optimal uptake of waste to meet their food requirements and increase survivability. Evidently, models have an important role to play and this study emphasizes the need for representative input data to describe actual food inputs, cage biomass changes, and management practices for more representative farm-scale modelling and essentially to improve particulate waste management. To effectively mitigate benthic impacts through IMTA, models must quantify and resolve particulate waste distribution and impact around fish farms to maintain a balanced system with net removal of wastes. Resolving farm-level complexities provides vital information about the variability of food availability and quality for extractive organisms that helps improve recycling of organic wastes in integrated systems, demanding a more representative modelling approach.

Abstract Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe β-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (β-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations.

We present a genome assembly from an individual male Philonthus cognatus (a rove beetle; Arthropoda; Insecta; Coleoptera; Staphylinidae). The genome sequence is 1,030.6 megabases in span. Most of the assembly is scaffolded into 12 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 20.7 kilobases in length. Gene annotation of this assembly on Ensembl identified 29,629 protein coding genes.

Abstract Global climate change introduces new combinations of environmental conditions, which is expected to increase stress on plants. This could affect many traits in multiple ways that are as yet unknown but will likely require the modification of existing genetic relationships among functional traits potentially involved in local adaptation. Theoretical evolutionary studies have determined that it is an advantage to have an excess of recombination events under heterogeneous environmental conditions. Our study, conducted on a population of radiata pine (Pinus radiata D. Don), was able to identify individuals that show high genetic recombination at genomic regions, which potentially include pleiotropic or collocating QTLs responsible for the studied traits, reaching a prediction accuracy of 0.80 in random cross-validation and 0.72 when whole family was removed from the training population and predicted. To identify these highly recombined individuals, a training population was constructed from correlation breakers, created through tandem selection of parents in the previous generation and their consequent mating. Although the correlation breakers showed lower observed heterogeneity possibly due to direct selection in both studied traits, the genomic regions with statistically significant differences in the linkage disequilibrium pattern showed higher level of heretozygosity, which has the effect of decomposing unfavourable genetic correlation. We propose undertaking selection of correlation breakers under current environmental conditions and using genomic predictions to increase the frequency of these ’recombined’ individuals in future plantations, ensuring the resilience of planted forests to changing climates. The increased frequency of such individuals will decrease the strength of the population-level genetic correlations among traits, increasing the opportunity for new trait combinations to be developed in the future.

Atmospheric deposition (AD) is a significant source of nutrient loading to waterbodies around the world. However, the sources and loading rates are poorly understood for major waterbodies and even less understood for local waterbodies. Utah Lake is a eutrophic lake located in central Utah, USA, and has high-nutrient levels. Recent research has identified AD as a significant source of nutrient loading to the lake, though contributions from dust particles make up 10% of total AD. To better understand the dust AD sources, we sampled suspected source locations and collected deposition samples around the lake. We analyzed these samples using Inductively Coupled Plasma (ICP) for 25 metals to characterize their elemental fingerprints. We then compared the lake samples to the source samples to determine likely source locations. We computed spectral angle, coefficient of determination, multi-dimensional scaling, and radar plots to characterize the similarity of the samples. We found that samples from local dust sources were more similar to dust in lake AD samples than samples from distant sources. This suggests that the major source of the dust portion of AD onto Utah Lake is the local empty fields south and west of the lake, and not the farther playa and desert sources as previously suggested. Preliminary data suggest that dust AD is associated with dry, windy conditions and is episodic in nature. We show that AD from dust particles is likely a small portion of the overall AD nutrient loading on Utah Lake, with the dry and precipitation sources contributing most of the load. This case identifies AD sources to Utah Lake and provides an example of data and methods that can be used to assess similarity or perform attribution for dust, soil, and other environmental data. While we use ICP metals, any number of features can be used with these methods if normalized.

Atmospheric deposition (AD) is a less understood and quantified source of nutrient loading to waterbodies. AD occurs via settling (large particulates), contact (smaller particulates and gaseous matter), and precipitation (rain, snow) transport pathways. Utah Lake is a shallow eutrophic freshwater lake located in central Utah, USA, with geophysical characteristics that make it particularly susceptible to AD-related nutrient loading. Studies have shown AD to be a significant contributor to the lake’s nutrient budget. This study analyzes nutrient samples from nine locations around the lake and four precipitation gauges over a 6-year study period using three different methods to estimate AD from the precipitation transport pathway. The methods used are simple averaging, Thiessen polygons, and inverse distance weighting, which we use to spatially interpolate point sample data to estimate nutrient lake loads. We hold that the inverse distance weighting method produces the most accurate results. We quantify, present, and compare nutrient loads and nutrient loading rates for total phosphorus (TP), total inorganic nitrogen (TIN), and ortho phosphate (OP) from precipitation events. We compute loading rates for the calendar year (Mg/yr) from each of the three analysis methods along with monthly loading rates where Mg is 106 g. Our estimated annual precipitation AD loads for TP, OP, and TIN are 120.96 Mg/yr (132.97 tons/yr), 60.87 Mg/yr (67.1 tons/yr), and 435 Mg/yr (479.5 tons/yr), respectively. We compare these results with published data on total AD nutrient loads and show that AD from precipitation is a significant nutrient source for Utah Lake, contributing between 25% and 40% of the total AD nutrient load to the lake.

Abstract Early detection of diseases such as COVID-19 could be a critical tool in reducing disease transmission by helping individuals recognize when they should self-isolate, seek testing, and obtain early medical intervention. Consumer wearable devices that continuously measure physiological metrics hold promise as tools for early illness detection. We gathered daily questionnaire data and physiological data using a consumer wearable (Oura Ring) from 63,153 participants, of whom 704 self-reported possible COVID-19 disease. We selected 73 of these 704 participants with reliable confirmation of COVID-19 by PCR testing and high-quality physiological data for algorithm training to identify onset of COVID-19 using machine learning classification. The algorithm identified COVID-19 an average of 2.75 days before participants sought diagnostic testing with a sensitivity of 82% and specificity of 63%. The receiving operating characteristic (ROC) area under the curve (AUC) was 0.819 (95% CI [0.809, 0.830]). Including continuous temperature yielded an AUC 4.9% higher than without this feature. For further validation, we obtained SARS CoV-2 antibody in a subset of participants and identified 10 additional participants who self-reported COVID-19 disease with antibody confirmation. The algorithm had an overall ROC AUC of 0.819 (95% CI [0.809, 0.830]), with a sensitivity of 90% and specificity of 80% in these additional participants. Finally, we observed substantial variation in accuracy based on age and biological sex. Findings highlight the importance of including temperature assessment, using continuous physiological features for alignment, and including diverse populations in algorithm development to optimize accuracy in COVID-19 detection from wearables.

Rowing ergometers are popular tools for general fitness and competitive crew teams. The effect of the equipment set up on the rowing stroke has received limited attention. This study aimed to determine the effects of altering the foot-stretcher position on rowing kinematics across different stroke rates. Eleven college-level rowers took part in this study. A rowing ergometer was modified to allow the height and angle of the foot-stretcher to be adjusted. Seven foot-stretcher positions were tested, each at rates of 22, 26, and 32 strokes per minute. Sagittal plane kinematic waveforms were compared between conditions for all major joints using statistical parametric mapping, and temporal variables were assessed (p < 0.05). Stroke rate was found to affect kinematic patterns for all joints. The effect of the foot-stretcher position was limited to the ankle and hip. Similarly, the timing of events during the rowing stroke was affected by the stroke rate, but not foot position. These results indicate that while some limited changes to the stroke technique can be caused by altering the foot-stretcher position, the changes were largely compensated for by the rowers and are generally smaller than differences between stroke rates.

Abstract Remote sensing of specific climatic and biogeographical parameters is an effective means of evaluating the large-scale desertification status of drylands affected by negative human impacts. Here, we identify and analyze desertification trends in Iran for the period 2001–2015 via a combination of three indices for vegetation (NPP—net primary production, NDVI—normalized difference vegetation index, LAI—leaf area index) and two climate indices (LST—land surface temperature, P—precipitation). We combine these indices to identify and map areas of Iran that are susceptible to land degradation. We then apply a simple linear regression method, the Mann–Kendall non-parametric test, and the Theil–Sen estimator to identify long-term temporal and spatial trends within the data. Based on desertification map, we find that 68% of Iran shows a high to very high susceptibility to desertification, representing an area of 1.1 million km2 (excluding 0.42 million km2 classified as unvegetated). Our results highlight the importance of scale in assessments of desertification, and the value of high-resolution data, in particular. Annually, no significant change is evident within any of the five indices, but significant changes (some positive, some negative) become apparent on a seasonal basis. Some observations follow expectations; for instance, NDVI is strongly associated with cooler, wet spring and summer seasons, and milder winters. Others require more explanation; for instance, vegetation appears decoupled from climatic forcing during autumn. Spatially, too, there is much local and regional variation, which is lost when the data are considered only at the largest nationwide scale. We identify a northwest–southeast belt spanning central Iran, which has experienced significant vegetation decline (2001–2015). We tentatively link this belt of land degradation with intensified agriculture in the hinterlands of Iran’s major cities. The spatial and temporal trends identified with the three vegetation and two climate indices afford a cost-effective framework for the prediction and management of future environmental trends in developing regions at risk of desertification.

Metal-organic framework adsorbents are promising materials for gas separation and purification. Herein, the authors present a metal-organic framework that selectively captures CO2 over small hydrocarbons; this separation is relevant for the purification of natural gas and industrial feedstocks.

In recent years, marine plastic pollution has seen increased coverage in the public interest and research due to a greater understanding of the scale and impact of plastic pollution within the marine environment. Considering the hazard that plastic waste poses on the environment, marine life, and on humans, remote-sensing techniques could provide timely information on their detection and dynamics. The remote sensing of marine plastic is a relatively new field and research into the capabilities of radar for detecting and monitoring marine plastic pollution is generally limited, with several interactions and mechanisms being largely unknown. Here, we exploit the use of a C- and X-band radar to understand the capabilities of monitoring marine plastics. Our results show that backscattering differences in the C- and X-band between the reference water (called here as “clean”) and the test water filled with plastic can be detected in some conditions (based on statistical analysis). Overall, the results indicate that the X-band frequency performs significantly better than the C-band frequency, with X-band detecting significant differences in backscattering in 48/68 test cases compared with C-band detecting differences in 20/67 test cases. We also find that the difference in backscattering is dependent on the size and shape of the plastic object, as well as the wave conditions which the plastic is moving on. This study provides new insights on the radar capabilities for detecting marine plastic litter and new information which can be used in the planning of future missions and studies on the remote sensing of marine plastic pollution.

Plastics in the river environment are of major concern due to their potential pathways into the ocean, their persistence in the environment, and their impacts on human and marine health. It has been documented that plastic concentrations in riparian environments are higher following major rain events, where plastic can be moved through surface runoff. Considering the hazard that plastic waste poses to the environment, monitoring techniques are needed to aid in locating, monitoring, and remediating plastic waste within these systems. Dams are known to trap sediments and pollutants, such as metals and Polychlorinated Biphenyls (PCBs). While there is an established background on the monitoring of dams using the synoptic coverage provided by satellite imaging to observe water quality and volume, the detection of marine debris in riparian systems remains challenging, especially in cloudy conditions. Herein, we exploit the use of Synthetic Aperture Radar (SAR) to understand its capabilities for monitoring marine debris. This research focuses on detecting plastic islands within the Drina River system in Bosnia and Herzegovina and Serbia. Here, the results show that the monitoring of these plastic accumulations is feasible using Sentinel-1 SAR data. A quantitative analysis of detection performance is presented using traditional and state-of-the-art change detectors. The analysis of these detectors indicates that detectors that can utilise the coherent data from Single Look Complex (SLC) acquisitions are perform better when compared with those that only utilise incoherent data from Ground Range-Detected (GRD) acquisitions, with true positive detection ratings of ~95% with 0.1% false alarm rates seen in the best-performing detector. We also found that that the cross-pol VH channel provides better detection than those based on single-pol VV polarisation.

Research on mutant tobacco plants shows that a pigment called β-carotene is not necessary for photosynthesis.

Metal–organic framework glasses have emerged as a new family of melt-quenched glass, but have yet to display the accessible porosity of their crystalline counterparts. Here, Bennett and colleagues report that glasses derived from ZIF-76 parent materials possess 4 – 8 Å pores and exhibit reversible gas adsorption.

Abstract The existence of a population of putative stem cells with germline developmental potential (oogonial stem cells: OSCs) in the adult mammalian ovary has been marked by controversy over isolation methodology and potential for in-vitro transformation, particularly where cell sorting has been based on expression of DEAD box polypeptide 4 (DDX4). This study describes a refined tissue dissociation/fluorescence-activated cell sorting (FACS) protocol for the ovaries of adult women which results in increased cell viability and yield of putative OSCs. A FACS technique incorporating dual-detection of DDX4 with aldehyde dehydrogenase 1 (ALDH1) demonstrates the existence of two sub-populations of small DDX4-positive cells (approx. 7 µm diameter) with ALDH1 activity, distinguished by expression of differentially spliced DDX4 transcripts and of DAZL, a major regulator of germ cell differentiation. These may indicate stages of differentiation from a progenitor population and provide a likely explanation for the expression disparities reported previously. These findings provide a robust basis for the further characterisation of these cells, and exploration of their potential physiological roles and therapeutic application.

Development of genome-wide resources for application in genomic selection or genome-wide association studies, in the absence of full reference genomes, present a challenge to the forestry industry, where longer breeding cycles could benefit from the accelerated selection possible through marker-based breeding value predictions. In particular, large conifer megagenomes require a strategy to reduce complexity, whilst ensuring genome-wide coverage is achieved. Using a transcriptome-based reference template, we have successfully developed a high density exome capture genotype-by-sequencing panel for radiata pine (Pinus radiata D.Don), capable of capturing in excess of 80,000 single nucleotide polymorphism (SNP) markers with a minor allele frequency above 0.03 in the population tested. This represents approximately 29,000 gene models from a core set of 48,914 probes. A set of 704 SNP markers capable of pedigree reconstruction and differentiating individual genotypes were tested within two full-sib mapping populations. While as few as 70 markers could reconstruct parentage in almost all cases, the impact of missing genotypes was noticeable in several offspring. Therefore, 60 sets of 110 randomly selected SNP markers were compared for both parentage reconstruction and clone differentiation. The performance in parentage reconstruction showed little variation over 60 iterations. However, there was notable variation in discriminatory power between closely related individuals, indicating a higher density SNP marker panel may be required to elucidate hidden relationships in complex pedigrees.

The pressure on scarce freshwater resources from intensifying human activity is rising across the globe. This study presents the case of Jatiluhur—Indonesia’s largest reservoir, where unregulated aquaculture expansion has contributed to environmental degradation and associated conflicts with other water-users. Aiming to identify a strategy to improve the sustainability of cage aquaculture within Jatiluhur reservoir and other freshwater bodies facing similar challenges, this study consisted of an initial analysis of time bound satellite images of the reservoir and a systematic survey of 112 aquaculture farms. The results revealed that, with more than 45 000 production units in 2020, more than half of which are placed outside government-approved aquaculture zones, the carrying capacity of the reservoir is exceeded. An analysis of the farm-level production practices, pertinent to environmental regulation, indicated the existence of three main production strategies, with significant differences in eutrophication potential among them that would lead to better articulated policy actions. A feed manufacturer-focused policy to reduce total phosphorus levels in formulated diets by more than half, but still within the optimal level for fish performance, would have the most impact. Enforcement of registration and removal of illegal production units supported by satellite-based monitoring of compliance are key recommendations to support intensive cage culture remaining an important economic activity.

Within the first year of implementation, 43% of women who tested HIV positive at their first antenatal care visit were no longer retained and being followed in the free prevention of mother to child transmission (PMTCT) of HIV program offered by the Kenyan Ministry of Health and Médecins Sans Frontières in the informal settlement of Kibera, Nairobi. This study aimed to explore barriers to enrolling and remaining engaged in PMTCT services throughout the pregnancy and postpartum periods. Qualitative data from 31 focus group discussions and 35 in-depth interviews across six stakeholder groups that included women, men, and PMTCT service providers were analyzed. Using an inductive exploratory approach, four researchers coded the data and identified key themes. Five themes emerged from the data that may influence attrition from PMTCT service in this setting: 1) HIV in the context of Kibera, 2) knowledge of HIV status, 3) knowledge of PMTCT, 4) disclosure of HIV status, and 5) male partner support for PMTCT services. A new HIV diagnosis during pregnancy immediately triggered an ongoing risk assessment of perceived hazards in the home, community, and clinic environments that could occur as a result of female participation in PMTCT services. Male partners were a major influence in this risk assessment, but were generally unaware of PMTCT services. To preserve relationships with male partners, meet community expectations of womanhood, and maintain confidentiality while following recommendations of healthcare providers, women had to continuously weigh the risks and benefits of PMTCT services and interventions. Community-based HIV testing and PMTCT education, male involvement in antenatal care, and counseling customized to assist each woman in her own unique risk assessment, may improve uptake of and retention in care and optimize the HIV prevention benefit of PMTCT interventions.

Glioblastoma (GBM) is the most common primary brain malignancy in adults, yet survival outcomes remain poor. First line treatment is well established, however disease invariably recurs and improving prognosis is challenging. With the aim of personalizing therapy at recurrence, we have established a high content screening (HCS) platform to analyze the sensitivity profile of seven patient-derived cancer stem cell lines to 83 FDA-approved chemotherapy drugs, with and without irradiation.Seven cancer stem cell lines were derived from patients with GBM and, along with the established cell line U87-MG, each patient-derived line was cultured in tandem in serum-free conditions as adherent monolayers and three-dimensional neurospheres. Chemotherapeutics were screened at multiple concentrations and cells double-stained to observe their effect on both cell death and proliferation. Sensitivity was classified using high-throughput algorithmic image analysis.Cell line specific drug responses were observed across the seven patient-derived cell lines. Few agents were seen to have radio-sensitizing effects, yet some drug classes showed a marked difference in efficacy between monolayers and neurospheres. In vivo validation of six drugs suggested that cell death readout in a three-dimensional culture scenario is a more physiologically relevant screening model and could be used effectively to assess the chemosensitivity of patient-derived GBM lines.The study puts forward a number of non-standard chemotherapeutics that could be useful in the treatment of recurrent GBM, namely mitoxantrone, bortezomib and actinomycin D, whilst demonstrating the potential of HCS to be used for personalized treatment based on the chemosensitivity profile of patient tumor cells.

There is a wide diversity of bioinformatic tools available for the assembly of next generation sequence and subsequence variant calling to identify genetic markers at scale. Integration of genomics tools such as genomic selection, association studies, pedigree analysis and analysis of genetic diversity, into operational breeding is a goal for New Zealand's most widely planted exotic tree species, Pinus radiata. In the absence of full reference genomes for large megagenomes such as in conifers, RNA sequencing in a range of genotypes and tissue types, offers a rich source of genetic markers for downstream application. We compared nine different assembler and variant calling software combinations in a single transcriptomic library and found that Single Nucleotide Polymorphism (SNPs) discovery could vary by as much as an order of magnitude (8,061 SNPs up to 86,815 SNPs). The assembler with the best realignment of the packages trialled, Trinity, in combination with several variant callers was then applied to a much larger multi-genotype, multi-tissue transcriptome and identified 683,135 in silico SNPs across a predicted 449,951 exons when mapped to the Pinus taeda ver 1.01e reference.

Weather conditions are commonly believed to influence musculoskeletal pain, however the evidence for this is mixed. This study aimed to examine the relationship between local meteorological conditions and online search trends for terms related to knee pain, hip pain, and arthritis. Five years of relative online search volumes for these terms were obtained for the 50 most populous cities in the contiguous United States, along with corresponding local weather data for temperature, relative humidity, barometric pressure, and precipitation. Methods from the climate econometrics literature were used to assess the casual impact of these meteorological variables on the relative volumes of searches for pain. For temperatures between -5°C and 30°C, search volumes for hip pain increased by 12 index points, and knee pain increased by 18 index points. Precipitation had a negative effect on search volumes for these terms. At temperatures >30°C, search volumes for arthritis related pain decreased by 7 index points. These patterns were not seen for pain searches unrelated to the musculoskeletal system. In summary, selected local weather conditions are significantly associated with online search volumes for specific musculoskeletal pain symptoms. We believe the predominate driver for this to be the relative changes in physical activity levels associated with meteorological conditions.

Accurate inference of relatedness between individuals in breeding population contributes to the precision of genetic parameter estimates, effectiveness of inbreeding management and the amount of genetic progress delivered from breeding programs. Pedigree reconstruction has been proven to be an efficient tool to correct pedigree errors and recover hidden relatedness in open pollinated progeny tests but the method can be limited by the lack of parental genotypes and the high proportion of alien pollen from outside the breeding population. Our study investigates the efficiency of sib-ship reconstruction in an advanced breeding population of Eucalyptus nitens with only partially tracked pedigree. The sib-ship reconstruction allowed the identification of selfs (4% of the sample) and the exploration of their potential effect on inbreeding depression in the traits studied. We detected signs of inbreeding depression in diameter at breast height and growth strain while no indications were observed in wood density, wood stiffness and tangential air-dry shrinkage. After the application of a corrected sib-ship relationship matrix, additive genetic variance and heritability were observed to increase where signs of inbreeding depression were initially detected. Conversely, the same genetic parameters for traits that appeared to be free of inbreeding depression decreased in size. It therefore appeared that greater genetic variance may be due, at least in part, to contributions from inbreeding in these studied populations rather than a removal of inbreeding as is traditionally thought.

Carbonic anhydrase IX (CAIX) is a tumor-specific protein that is upregulated during hypoxic conditions where it is involved in maintaining the pH balance. CAIX causes extracellular acidification, thereby limiting the uptake of weak basic chemotherapeutic agents, such as doxorubicin, and decreasing its efficacy. The aim of this study was to determine if doxorubicin efficacy can be increased when combined with the selective sulfamate CAIX inhibitor S4. The effect of S4 on doxorubicin efficacy was tested in vitro using cell viability assays with MDA-MB-231, FaDu, HT29 -CAIX high and HT29 -CAIX low cell lines. In addition, the efficacy of this combination therapy was investigated in tumor xenografts of the same cell lines. The addition of S4 in vitro increased the efficacy of doxorubicin in the MDA-MB-231 during hypoxic exposure (IC50 is 0.25 versus 0.14 µM, p = 0.0003). Similar results were observed for HT29-CAIX high with S4 during normoxia (IC50 is 0.20 versus 0.08 µM, p

Pedigree reconstruction using molecular markers enables efficient management of inbreeding in open-pollinated breeding strategies, replacing expensive and time-consuming controlled pollination. This is particularly useful in preferentially outcrossed, insect pollinated Eucalypts known to suffer considerable inbreeding depression from related matings. A single nucleotide polymorphism (SNP) marker panel consisting of 106 markers was selected for pedigree reconstruction from the recently developed high-density Eucalyptus Infinium SNP chip (EuCHIP60K). The performance of this SNP panel for pedigree reconstruction in open-pollinated progenies of two Eucalyptus nitens seed orchards was compared with that of two microsatellite panels with 13 and 16 markers respectively. The SNP marker panel out-performed one of the microsatellite panels in the resolution power to reconstruct pedigrees and out-performed both panels with respect to data quality. Parentage of all but one offspring in each clonal seed orchard was correctly matched to the expected seed parent using the SNP marker panel, whereas parentage assignment to less than a third of the expected seed parents were supported using the 13-microsatellite panel. The 16-microsatellite panel supported all but one of the recorded seed parents, one better than the SNP panel, although there was still a considerable level of missing and inconsistent data. SNP marker data was considerably superior to microsatellite data in accuracy, reproducibility and robustness. Although microsatellites and SNPs data provide equivalent resolution for pedigree reconstruction, microsatellite analysis requires more time and experience to deal with the uncertainties of allele calling and faces challenges for data transferability across labs and over time. While microsatellite analysis will continue to be useful for some breeding tasks due to the high information content, existing infrastructure and low operating costs, the multi-species SNP resource available with the EuCHIP60k, opens a whole new array of opportunities for high-throughput, genome-wide or targeted genotyping in species of Eucalyptus.

Therapeutic inhibition of poly(ADP-ribose) polymerase (PARP), as monotherapy or to supplement the potencies of other agents, is a promising strategy in cancer treatment. We previously reported that the first PARP inhibitor to enter clinical trial, rucaparib (AG014699), induced vasodilation in vivo in xenografts, potentiating response to temozolomide. We now report that rucaparib inhibits the activity of the muscle contraction mediator myosin light chain kinase (MLCK) 10-fold more potently than its commercially available inhibitor ML-9. Moreover, rucaparib produces additive relaxation above the maximal degree achievable with ML-9, suggesting that MLCK inhibition is not solely responsible for dilation. Inhibition of nitric oxide synthesis using L-NMMA also failed to impact rucaparib's activity. Rucaparib contains the nicotinamide pharmacophore, suggesting it may inhibit other NAD+-dependent processes. NAD+ exerts P2 purinergic receptor-dependent inhibition of smooth muscle contraction. Indiscriminate blockade of the P2 purinergic receptors with suramin abrogated rucaparib-induced vasodilation in rat arterial tissue without affecting ML-9-evoked dilation, although the specific receptor subtypes responsible have not been unequivocally identified. Furthermore, dorsal window chamber and real time tumor vessel perfusion analyses in PARP-1-/- mice indicate a potential role for PARP in dilation of tumor-recruited vessels. Finally, rucaparib provoked relaxation in 70% of patient-derived tumor-associated vessels. These data provide tantalising evidence of the complexity of the mechanism underlying rucaparib-mediated vasodilation.

BackgroundTo improve retention on ART, Médecins Sans Frontières, the Ministry of Health and patients piloted a community-based antiretroviral distribution and adherence monitoring model through Community ART Groups (CAG) in Tete, Mozambique. By December 2012, almost 6000 patients on ART had formed groups of whom 95.7% were retained in care. We conducted a qualitative study to evaluate the relevance, dynamic and impact of the CAG model on patients, their communities and the healthcare system.MethodsBetween October 2011 and May 2012, we conducted 16 focus group discussions and 24 in-depth interviews with the major stakeholders involved in the CAG model. Audio-recorded data were transcribed verbatim and analysed using a grounded theory approach.ResultsSix key themes emerged from the data: 1) Barriers to access HIV care, 2) CAG functioning and actors involved, 3) Benefits for CAG members, 4) Impacts of CAG beyond the group members, 5) Setbacks, and 6) Acceptance and future expectations of the CAG model. The model provides cost and time savings, certainty of ART access and mutual peer support resulting in better adherence to treatment. Through the active role of patients, HIV information could be conveyed to the broader community, leading to an increased uptake of services and positive transformation of the identity of people living with HIV. Potential pitfalls included limited access to CAG for those most vulnerable to defaulting, some inequity to patients in individual ART care and a high dependency on counsellors.ConclusionThe CAG model resulted in active patient involvement and empowerment, and the creation of a supportive environment improving the ART retention. It also sparked a reorientation of healthcare services towards the community and strengthened community actions. Successful implementation and scalability requires (a) the acceptance of patients as partners in health, (b) adequate resources, and (c) a well-functioning monitoring and management system.

Over the past two decades finite element (FE) analysis has become a popular tool for researchers seeking to simulate the biomechanics of the healthy and diabetic foot. The primary aims of these simulations have been to improve our understanding of the foot's complicated mechanical loading in health and disease and to inform interventions designed to prevent plantar ulceration, a major complication of diabetes. This article provides a systematic review and summary of the findings from FE analysis-based computational simulations of the diabetic foot.A systematic literature search was carried out and 31 relevant articles were identified covering three primary themes: methodological aspects relevant to modelling the diabetic foot; investigations of the pathomechanics of the diabetic foot; and simulation-based design of interventions to reduce ulceration risk.Methodological studies illustrated appropriate use of FE analysis for simulation of foot mechanics, incorporating nonlinear tissue mechanics, contact and rigid body movements. FE studies of pathomechanics have provided estimates of internal soft tissue stresses, and suggest that such stresses may often be considerably larger than those measured at the plantar surface and are proportionally greater in the diabetic foot compared to controls. FE analysis allowed evaluation of insole performance and development of new insole designs, footwear and corrective surgery to effectively provide intervention strategies. The technique also presents the opportunity to simulate the effect of changes associated with the diabetic foot on non-mechanical factors such as blood supply to local tissues.While significant advancement in diabetic foot research has been made possible by the use of FE analysis, translational utility of this powerful tool for routine clinical care at the patient level requires adoption of cost-effective (both in terms of labour and computation) and reliable approaches with clear clinical validity for decision making.

The transcription factors Runx1 and c-Myc have individually been shown to regulate important gene targets as well as to collaborate in oncogenesis. However, it is unknown whether there is a regulatory relationship between the two genes. In this study, we investigated the transcriptional regulation of endogenous c-Myc by Runx1 in the human T cell line Jurkat and murine primary hematopoietic cells. Endogenous Runx1 binds to multiple sites in the c-Myc locus upstream of the c-Myc transcriptional start site. Cells transduced with a C-terminally truncated Runx1 (Runx1.d190), which lacks important cofactor interaction sites and can block C-terminal-dependent functions of all Runx transcription factors, showed increased transcription of c-Myc. In order to monitor c-Myc expression in response to early and transiently-acting Runx1.d190, we generated a cell membrane-permeable TAT-Runx1.d190 fusion protein. Murine splenocytes treated with TAT-Runx1.d190 showed an increase in the transcription of c-Myc within 2 hours, peaking at 4 hours post-treatment and declining thereafter. This effect is dependent on the ability of Runx1.d190 to bind to DNA. The increase in c-Myc transcripts is correlated with increased c-Myc protein levels. Collectively, these data show that Runx1 directly regulates c-Myc transcription in a C-terminal- and DNA-binding-dependent manner.

Plague is endemic within the central highlands of Madagascar, where its main reservoir is the black rat, Rattus rattus. Typically this species is considered susceptible to plague, rapidly dying after infection inducing the spread of infected fleas and, therefore, dissemination of the disease to humans. However, persistence of transmission foci in the same area from year to year, supposes mechanisms of maintenance among which rat immune responses could play a major role. Immunity against plague and subsequent rat survival could play an important role in the stabilization of the foci. In this study, we aimed to investigate serological responses to plague in wild black rats from endemic areas of Madagascar. In addition, we evaluate the use of a recently developed rapid serological diagnostic test to investigate the immune response of potential reservoir hosts in plague foci.We experimentally infected wild rats with Yersinia pestis to investigate short and long-term antibody responses. Anti-F1 IgM and IgG were detected to evaluate this antibody response. High levels of anti-F1 IgM and IgG were found in rats one and three weeks respectively after challenge, with responses greatly differing between villages. Plateau in anti-F1 IgM and IgG responses were reached for as few as 500 and 1500 colony forming units (cfu) inoculated respectively. More than 10% of rats were able to maintain anti-F1 responses for more than one year. This anti-F1 response was conveniently followed using dipsticks.Inoculation of very few bacteria is sufficient to induce high immune response in wild rats, allowing their survival after infection. A great heterogeneity of rat immune responses was found within and between villages which could heavily impact on plague epidemiology. In addition, results indicate that, in the field, anti-F1 dipsticks are efficient to investigate plague outbreaks several months after transmission.

The gastrointestinal tract microbiota (GTM) of mammals is a complex microbial consortium, the composition and activities of which influences mucosal development, immunity, nutrition and drug metabolism. It remains unclear whether the composition of the dominant GTM is conserved within animals of the same strain and whether stable GTMs are selected for by host-specific factors or dictated by environmental variables.The GTM composition of six highly inbred, genetically distinct strains of mouse (C3H, C57, GFEC, CD1, CBA nu/nu and SCID) was profiled using eubacterial -specific PCR-DGGE and quantitative PCR of feces. Animals exhibited strain-specific fecal eubacterial profiles that were highly stable (c. >95% concordance over 26 months for C57). Analyses of mice that had been relocated before and after maturity indicated marked, reproducible changes in fecal consortia and that occurred only in young animals. Implantation of a female BDF1 mouse with genetically distinct (C57 and Agoutie) embryos produced highly similar GTM profiles (c. 95% concordance) between mother and offspring, regardless of offspring strain, which was also reflected in urinary metabolite profiles. Marked institution-specific GTM profiles were apparent in C3H mice raised in two different research institutions.Strain-specific data were suggestive of genetic determination of the composition and activities of intestinal symbiotic consortia. However, relocation studies and uterine implantation demonstrated the dominance of environmental influences on the GTM. This was manifested in large variations between isogenic adult mice reared in different research institutions.