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Only one virus, Avipox, has been documented previously in wild birds in Hawaii. Using immunohistochemistry and PCR, we found that two native threatened Hawaiian Geese (Branta sandvicensis), one with multicentric histiocytoma and the other with toxoplasmosis, and one Laysan Albatross (Phoebastria immutabilis) with avian pox were infected with reticuloendotheliosis virus (REV). The virus was isolated from one of the geese by cell culture. Surveys of other Hawaiian geese with various pathologies, avian pox cases, and pox viral isolates using PCR failed to reveal REV, suggesting that the virus is uncommon, at least in samples examined. The full genome of the Gag, Pol, and Env genes were sequenced for all three infected birds and revealed geographic divergence of the Pol gene, suggesting it to be under strong selective pressure. Our finding of REV in Hawaii makes this only the second virus documented in native Hawaiian birds associated with pathology. Moreover, the presence of REV in a pelagic seabird is unusual. Future surveys should seek the reservoir of the virus in efforts to trace its origins.

Aluminium (Al) is highly toxic to plant growth, with soluble concentrations being elevated in the ∼40% of arable soils worldwide that are acidic. Determining the distribution of Al in plant tissues is important for understanding the mechanisms by which it is toxic and how some plants tolerate high concentrations. Synchrotron- and laboratory-based X-ray fluorescence microscopy (XFM) is a powerful technique to quantitatively analyse the distribution of elements, including in hydrated and living plants. However, analysis of light elements (z < phosphorus) is extremely challenging due to signal losses in air, and the unsuitability of vacuum environments for (fresh) hydrated plant tissues. This study uses XFM in a helium environment to avoid Al signal loss to reveal the distribution of Al in hydrated plant tissues of Tea (Camellia sinensis). The results show that Al occurs in localised areas across the foliar surface, whereas in cross-sections Al is almost exclusively concentrated in the apoplastic space above and in between adaxial epidermal cells. This distribution of Al is related to the Al tolerance of this species, and accumulation of phytotoxic elements in the apoplastic space, away from sensitive processes such as photosynthesis in the palisade mesophyll cells, is a common tolerance mechanism reported in many different plant species. This study develops an XFM method on both synchrotron and laboratory sources that overcomes the drawbacks of existing analytical techniques, permitting measurement of light elements down to Al in (fresh) hydrated plant tissues.

The flora of Papua New Guinea is amongst the richest in the world with an estimated 25,000 plant species. The extreme levels of biodiversity, climatic ranges and soil types suggest a high possibility of metal hyperaccumulator plants existing in Papua New Guinea. However, no hyperaccumulator plants have been reported from this region yet. The use of handheld X-ray fluorescence instruments is a non-destructive and effective method for the systematic quantitative assessment of hyperaccumulation in vast numbers of herbarium specimens. X-ray fluorescence scanning was undertaken at the Queensland Herbarium (Australia) on all Papua New Guinea specimens from seven major families (Celastraceae, Cunoniaceae, Phyllanthaceae, Proteaceae, Rubiaceae, Salicaceae and Violaceae), covering 3164 plant specimens. This preliminary survey revealed the existence of ten zinc hyperaccumulator species (> 3000 µg g−1 Zn), eight manganese accumulator species (> 5000 µg g−1 Mn) and one nickel hyperaccumulator species (> 1000 µg g−1 Ni). These results highlight the potential for discovery of numerous new metal hyperaccumulator plants from the flora of Papua New Guinea if larger-scale systematic screening efforts were undertaken.

BackgroundHyperaccumulation of trace elements is a rare trait among plants which is being investigated to advance our understanding of the regulation of metal accumulation and applications in phytotechnologies.Noccaea caerulescens(Brassicaceae) is an intensively studied hyperaccumulator model plant capable of attaining extremely high tissue concentrations of zinc and nickel with substantial genetic variation at the population-level. Micro-X-ray Fluorescence spectroscopy (µXRF) mapping is a sensitive high-resolution technique to obtain information of the spatial distribution of the plant metallome in hydrated samples. We used laboratory-based µXRF to characterize a collection of 86 genetically diverseNoccaea caerulescensaccessions from across Europe. We developed an image-processing method to segment different plant substructures in the µXRF images. We introduced the concentration quotient (CQ) to quantify spatial patterns of metal accumulation and linked that to genetic variation.ResultsImage processing resulted in automated segmentation of µXRF plant images into petiole, leaf margin, leaf interveinal and leaf vasculature substructures. The harmonic means of recall and precision (F1 score) were 0.79, 0.80, 0.67, and 0.68, respectively. Spatial metal accumulation as determined by CQ is highly heritable inNoccaea caerulescensfor all substructures, with broad-sense heritability (H2) ranging from 76 to 92%, and correlates only weakly with other heritable traits. Insertion of noise into the image segmentation algorithm barely decreases heritability scores of CQ for the segmented substructures, illustrating the robustness of the trait and the quantification method. Very low heritability was found for CQ if randomly generated substructures were compared, validating the approach.ConclusionsA strategy for segmenting µXRF images ofNoccaea caerulescensis proposed and the concentration quotient is developed to provide a quantitative measure of metal accumulation pattern, which can be used to determine genetic variation for such pattern. The metric is robust to segmentation error and provides reliable H2estimates. This strategy provides an avenue for quantifying XRF data for analysis of the genetics of metal distribution patterns in plants and the subsequent discovery of new genes that regulate metal homeostasis and sequestration in plants.

NAD depletion as pathogen response One way that plants respond to pathogen infection is by sacrificing the infected cells. The nucleotide-binding leucine-rich repeat immune receptors responsible for this hypersensitive response carry Toll/interleukin-1 receptor (TIR) domains. In two papers, Horsefield et al. and Wan et al. report that these TIR domains cleave the metabolic cofactor nicotinamide adenine dinucleotide (NAD + ) as part of their cell-death signaling in response to pathogens. Similar signaling links mammalian TIR-containing proteins to NAD + depletion during Wallerian degeneration of neurons. Science , this issue p. 793 , p. 799

Researchers are finding merits in utilizing industry-specific safety climate scales that capture the nuances of context, and tend to show stronger associations with safety behavior and outcomes like incidents. Yet, to date, guidance around the practicalities of developing and validating such industry-specific scales is lacking in the safety science literature.In this paper we outline our experiences developing six industry-specific safety climate scales and highlight strengths and limitations of our approach. We also briefly review the industry-specific safety climate literature and offer highlights for consideration when developing such scales. Our method to develop industry-specific safety climate scales followed an established best practice structure: literature review of existing published industry scales, collation and review of existing scale items, consultation interviews with industry experts, item drafting, exploratory and confirmatory statistical analyses, and finally, a real-world ecological validity test.Our research highlighted the diversity of safety climate dimensions (both the conceptual and content domains of each dimension) when it is considered at an industry level. Also, the literature reviews revealed a dearth of industry-specific safety climate scales in the areas we engaged with, so our project filled a glaring gap in research and practice. Best practice safety climate scale development methods are provided to stimulate further research.We conclude with reflections on the nature of safety climate within and across industries, and offer suggestions for future lines of research across other contexts (e.g., national culture, geography, and regulatory settings). We suggest that industry-specific safety climate scales have a specific use case, such as identifying specific areas to improve and evaluating the impact of safety interventions.This article provides applications for both applied researchers (to improve capabilities in safety climate scale development) and practitioners who wish to measure organisational safety climate and design effective interventions. Engaging with regulators to build safety climate scales is powerful because their personnel have rich experiences to share across multiple workplaces. Organisational researchers can engage with survey panels to build robust scales. Finally, industry-specific nuances can lead to richer insights into an organisation's safety climate.

Background During the coronavirus pandemic, it was essential to ensure access to End of life (EoL) medications while protecting the medications supply chain. Traditional Just in Case prescribing could have exhausted the national supply. Consequently, a new NHS Wales Interim End of Life COVID-19 Medicines Service was implemented to provide access to a medication box a Just-In-Time Emergency Medication Pack (JEMP). Methods The JEMP scheme ensured that end of life medications could be delivered directly to the patient within 2 hours, 24 hours a day 7 days a week, across the whole of Wales. The JEMP contained an agreed list of medications. The Health Courier Service Wales managed a Single Point of Contact, and directed the JEMP request to the most appropriate EoL Pharmacy Hub situated at one of five strategic sites around Wales. To access the service, a healthcare professional would call a 24/7 phone line and send a digital image of the prescription for EoL medications via Hospify (like an NHS Whatsapp). The JEMP then would be delivered directly to the patient‘s house and the prescription would be collected. Results 17 requests were activated. 1 call was stood down. 15 out of the 16 requests had a JEMP delivered within 2 hours. This ranged from 34 minutes to 111 minutes. The mean time was 72 minutes. Conclusion This is a pioneering service that developed during the coronavirus pandemic, it went from concept to a national pilot in 6 days. The uptake of services was less than originally predicted. However, it has been successful in its primary aims to maintain the supply chain and deliver vital EoL medications directly to the patient anywhere in Wales (24 hours 7 days a week) within 2 hours from the initial request. Further work needs to be done on collecting feedback and assessing sustainability.

We describe an undocumented change in how calm periods in near-surface wind speed (and direction) observations have been encoded in a subset of global datasets of sub-daily data after 2013. This has resulted in the under-estimation of the number of calm periods for meteorological stations across much of Asia and Europe. Hence average wind speeds after 2013 have been over-estimated, affecting the assessment of changes in global stilling and reversal phenomena after this date. By addressing this encoding change we show that globally, since 2010, wind speeds have recovered by around 30% less than previously thought., ZZ was supported by the National Natural Science Foundation of China (grants no. 42071022). CAM was supported by the Spanish Ministry of Science and Innovation (RTI2018-095749-A-I00), the Valencian Regional Government (AICO/2021/023), the Leonardo grant 2021 from the BBVA Foundation, and the Interdisciplinary Thematic Platform PTI-CLIMA. CS was supported by Swedish Formas(2019-00509 and 2017-01408) and VR (2021-02163 and 2019-03954).

Background Overexpression of CD47 by tumor cells exploits an immune checkpoint preventing tumor recognition and destruction by innate immune cells. Binding of tumor CD47 to SIRPα on macrophages and dendritic cells triggers a ‘don’t eat me’ signal that inhibits phagocytosis and allows escape from innate immune surveillance. Blockade of the CD47/SIRPα axis, however, enables immune recognition and phagocytic clearance of tumor cells. We have developed a clinical stage CD47 targeting antibody AO-176 that is highly differentiated among agents in this class. AO-176 not only blocks the CD47/SIRPα interaction and induces phagocytosis of tumor cells, but it also has a direct killing mechanism (via PCDIII) and induction of immunogenic cell death, leveraged by preferential binding to tumor versus normal cell CD47. Methods CD47 and β1 integrin expression and localization were evaluated using a combination of flow cytometry, western blotting, confocal microscopy and immunohistochemistry. Results Previously, we described that the preferential binding of AO-176 to tumor versus normal cells was due to its interaction with CD47 molecules that were pre-complexed to β1 integrin. This finding was particularly important and suggestive of why AO-176 does not bind red blood cells since they do not express β1 integrin. We have extended these findings to show that β1 integrin as well as CD47 are also expressed at lower levels in normal versus tumor cells, and that solid and hematologic tumor cells overexpress both CD47 and β1 integrin which correlate with poor prognosis in cancer. In addition, we show that AO-176 is able to bind and occupy CD47/β1 integrin complexes to a greater extent at acidic versus physiologic pH such as would be found in tumor microenvironments, an observation that also contributes to the enhanced targeting of AO-176 to tumor cells. Taken together, these findings add further insight into the preferential binding of AO-176 to tumor versus normal cells. Conclusions The context dependent binding of AO-176 to CD47, when complexed to β1 integrin, is unique among CD47 axis targeting agents and together with its direct killing mechanism of action offers a potentially better safety profile and opportunity for a therapeutic advantage. AO-176 is currently being evaluated in Phase 1 clinical trials for the treatment of patients with select solid tumors (NCT03834948) and multiple myeloma (NCT04445701). Trial Registration NCT03834948, NCT04445701.

Selenium (Se), a trace element essential for human and animal biological processes, is deficient in many agricultural soils. Some extremely rare plants can naturally accumulate extraordinarily high concentrations of Se. The native legume Neptunia amplexicaulis, endemic to a small area near Richmond and Hughenden in Central Queensland, Australia, is one of the strongest Se hyperaccumulators known on Earth, with foliar concentrations in excess of 4000 μg Se g−1 previously recorded. Here, we report on the Se distribution at a whole plant level using laboratory micro X-ray Fluorescence Microscopy (μXRF) and scanning electron microscopy (SEM-EDS), as well as on chemical forms of Se in various tissues using liquid chromatography-mass spectrometry (LC-MS) and synchrotron X-ray absorption spectroscopy (XAS). The results show that Se occurs in the forms of methyl-selenocysteine and seleno-methionine in the foliar tissues, with up to 13 600 μg Se g−1 total in young leaves. Selenium was found to accumulate primarily in the young leaves, flowers, pods and taproot, with lower concentrations present in the fine-roots and stem and the lowest present in the oldest leaves. Trichomes were not found to accumulate Se. We postulate that Se is (re)distributed in this plant via the phloem from older leaves to newer leaves, using the taproot as the main storage organ. High concentrations of Se in the nodes (pulvini) indicate this structure may play an important a role in Se (re)distribution. The overall pattern of Se distribution was similar in a non-Se tolerant closely related species (Neptunia gracilis), although the prevailing Se concentrations were substantially lower than in N. amplexicaulis.

This work investigates how one can postprocess a series of flash-generated X-ray radiographs with known point spread functions and collection geometries to produce radiographs with enhanced image quality and/or to approximate a multiframe X-ray movie by spatially correlating temporally sequenced images. To produce images with enhanced quality, one collects multiple individual projections, simultaneously, and then uses the known point spread function and acquisition geometry to correlate and integrate the numerous projections as if they were acquired from approximately the same perspective. If using sources of similar spectral characteristics, the outcome correlates with that which would be produced from a multiflash integration, where one benefits from an increase in flux of X-ray photons. If using sources of different spectral characteristics, a composite radiograph can be produced where the unique source spectrum can be matched to known material absorption cross-sections to accentuate radiographic features within the target material. To produce multiframe movies, similar mathematics are applied to images collected throughout a timespan over which the object changes. The aforementioned processes’ mathematics and examples are demonstrated for the case in which the individual projections are completely separable on the detector with no overlap. Finally, a potential method is discussed for separation of images in the case where multiple radiograph projections are overlapped and entangled on the detector.

The objective of this study was to evaluate the performance of two commercial‐grade electromagnetic sensors in measuring soil water content for the U.S. Climate Reference Network (USCRN). One sensor was a 50‐MHz coaxial impedance dielectric sensor (model HydraProbe, Stevens Water Monitoring Systems). The second sensor was a 1‐GHz time domain reflectometry (TDR) sensor (model TDR‐315L, Acclima). There was no substantial difference between both probes in daily mean volumetric soil water content and soil temperature measurements at a 0.1‐m depth in a customized uniform loamy soil testbed in Oak Ridge, TN, from 2016 to 2018, and at select USCRN stations with a variety of soils. However, the TDR‐315L provided more representative soil water content measurements for stations with high‐clay‐content soils than the HydraProbe. These results confirm the necessity of using site‐specific soil properties for soil sensors like the HydraProbe for estimating soil water content in different soil environments. They also suggest that TDR sensors may provide an opportunity to improve USCRN soil moisture measurements in higher‐clay‐content soils.

Background and Aims Synchrotron- and laboratory-based micro-X-ray fluorescence (µ-XRF) is a powerful technique to quantify the distribution of elements in physically large intact samples, including live plants, at room temperature and atmospheric pressure. However, analysis of light elements with atomic number (Z) less than that of phosphorus is challenging due to the need for a vacuum, which of course is not compatible with live plant material, or the availability of a helium environment. Method A new laboratory µ-XRF instrument was used to examine the effects of silicon (Si) on the manganese (Mn) status of soybean (Glycine max) and sunflower (Helianthus annuus) grown at elevated Mn in solution. The use of a helium environment allowed for highly sensitive detection of both Si and Mn to determine their distribution. Key Results The µ-XRF analysis revealed that when Si was added to the nutrient solution, the Si also accumulated in the base of the trichomes, being co-located with the Mn and reducing the darkening of the trichomes. The addition of Si did not reduce the concentrations of Mn in accumulations despite seeming to reduce its adverse effects. Conclusions The ability to gain information on the dynamics of the metallome or ionome within living plants or excised hydrated tissues can offer valuable insights into their ecophysiology, and laboratory µ-XRF is likely to become available to more plant scientists for use in their research.

A new optimized size exclusion chromatography small-angle X-ray scattering (SEC-SAXS) system for biomolecular SAXS at the Australian Synchrotron SAXS/WAXS beamline has been developed. The compact configuration reduces sample dilution to maximize sensitivity. Coflow sample presentation allows an 11-fold increase in flux on sample without capillary fouling, improving throughput and data quality, which are now primarily limited by the full flux available on the beamline. Multi-wavelength fibre optic UV analysis in close proximity to the X-ray beam allows for accurate concentration determination for samples with known UV extinction coefficients and thus estimation of the molecular weight of the scattering particle from the forward X-ray scattering intensity. Fast-flow low-volume SEC columns provide sample throughput competitive with batch concentration series measurements, albeit with a concomitant reduction of potential resolution relative to lower flow rates and larger SEC columns. The performance of the system is demonstrated using a set of model proteins, and its utility to solve various challenges is illustrated with a diverse suite of protein samples. These developments increase the quality and rigor of SEC-SAXS analysis and open new avenues for biomolecular solution SEC-SAXS studies that have been challenged by low sample yields, temporal instability, radiation sensitivity and complex mixtures.

Genome sequencing efforts have identified virtually all of the important mutations in adult myeloid malignancies. More recently, population studies have identified cancer-associated variants in the blood of otherwise healthy individuals as they age, a phenomenon termed clonal hematopoiesis of indeterminate potential (CHIP). This suggests that these mutations may occur in hematopoietic stem cells (HSCs) long before any clinical presentation but are not necessarily harbingers of transformation because only a fraction of individuals with CHIP develop hematopoietic pathologies. Delineation between CHIP variants that predispose for disease versus those that are more benign could be used as a prognostic factor to identify individuals at greater risk for transformation. To achieve this, the biological impact of CHIP variants on HSC function must be validated. One variant that has been identified recurrently in CHIP is a gain-of-function missense mutation in the imprinted gene GNAS (Guanine Nucleotide Binding Protein, Alpha Stimulating). In this study, we examined the effect of the GNASR201C variant on HSC function. Ectopic expression of GNASR201C supported transplantable HSC activity and improved lymphoid output in secondary recipients. Because declining lymphoid output is a hallmark of aging, GNASR201C mutations may sustain lymphoid-biased HSCs over time and maintain them in a developmental state favorable for transformation.

Turfgrass renovations commonly involve changing cultivars or species that are better suited for a given setting. Common bermudagrass [Cynodon dactylon (L.) Pers.] is a perennial turfgrass that is difficult to eradicate before renovations, and poses contaminant concerns for the subsequent stand. Dazomet is a granular soil fumigant that has activity on various pests, including common bermudagrass. Field research was conducted from 2015 to 2016 in Raleigh, NC and College Station, TX to evaluate dazomet treatments including various combinations of soil incorporation (irrigation- or tillage-incorporated) and sealing (tarp or no tarp) methods, application rates [291, 291 followed by (fb) 291, 468, or 583 kg·ha−1], and fluazifop-P [fluazifop (0.4 kg·ha−1)] + glyphosate (2.8 kg·ha−1 acid equivalent) application(s) for established common bermudagrass control. Overall, treatments required fluazifop + glyphosate before dazomet application for acceptable control (>90% cover reduction) at 42 and 46 weeks after initial treatment (WAIT) in Texas and North Carolina, respectively. Soil-incorporation results varied by location, with dazomet application (583 kg·ha−1) fb tillage resulting in ≥88% cover reduction across locations, while acceptable control from irrigation incorporation was only observed in North Carolina. Tarping did not improve efficacy when tillage incorporation at the maximum label application rate provided acceptable control, suggesting practitioners may eliminate this procedure. Information from this research will aid turfgrass managers in developing cost-effective, ecologically sound common bermudagrass eradication programs before renovations.

Hypervelocity impacts of copper rods into liquid filled channels circumferentially confined by steel cylinders were investigated using 2D axisymmetric hydrocode simulations. The confined liquid was utilized as a means to investigate the interplay of eroded target material and the remaining projectile in order to elucidate relevant penetration behaviors in solid materials. This led to a better understanding of two significant contributing parameters in the flow characteristics: the density (how difficult the material is to accelerate) and the channel diameter (how far the material has to move). While the liquid eliminates direct strength effects on penetration of the rods, varying the channel width allows a survey of parameter space that is typically a function of both material strength and density. Three successive copper rods with varying velocities and delays were used to model a penetrator representative of select sections of a shaped charge jet. For each target material, the residual penetration in solid rolled homogeneous armor (RHA) was then measured as a function of the liquid channel width. A strong dependence on both the material density and channel width was observed in regards to the efficiency of the material against penetration.

While T cell checkpoint inhibitors are mainstays of cancer immunotherapy, therapies that direct innate immune responses against cancer are lacking. CD47, a "don't eat me" signal, is an innate immune cell checkpoint which binds SIRPα on macrophages and dendritic cells to limit phagocytosis and its upregulation on tumor cells leads to evasion of immune detection and clearance. Therapeutic antibodies have previously been developed to block CD47 and induce phagocytosis of tumor cells, thus validating the pathway. AO-176, a next generation humanized IgG2 anti-CD47 antibody, was developed to block the CD47/SIRPα interaction and induce tumor cell phagocytosis. Moreover, AO-176 directly kills tumor cells through a non-ADCC-dependent mechanism via induction of programmed cell death type III. In addition to these tumor eliminating properties, AO-176 has the potential for a strong safety profile as a result of its preferential binding to tumor versus normal cells, lack of RBC binding, and enhanced binding to tumor cells at acidic pH. CD47 has previously been shown to be upregulated on acute myeloid leukemia (AML) leukemic stem cells (LSC), enabling their expansion through evasion from phagocytic clearance. As a result, patients with increased CD47 on AML LSCs have worse overall survival. In this study, AO-176 efficacy was evaluated in AML cell lines as a single agent and in combination with azacitidine and venetoclax which are approved therapies for AML. Azacitidine is a cytosine analogue which acts to inhibit DNA methylation, and venetoclax is a potent Bcl-2 inhibitor. Previous studies have shown that azacitidine induces apoptosis of tumor cells and increases cell surface exposure of calreticulin, a DAMP (Damage Associate Molecular Pattern) which provides a strong pro-phagocytic signal. From these findings, it was hypothesized that azacitidine would enable increased tumor cell phagocytosis when combined with AO-176. The potential for a similar enhancement with a combination of AO-176 and venetoclax was also explored. The ability of AO-176, with or without azacitidine or venetoclax, to induce DAMPs on the surface of AML cells was assessed. Cell surface expression of DAMPs, calreticulin and PDIA3, were measured by flow cytometry. AO-176, azacitidine, and venetoclax as single agents potently increased both calreticulin and PDIA3 in a dose-dependent manner on AML cell lines such as HL60 This is the first time, to our knowledge, that venetoclax has been shown to induce DAMPs. To better understand the functional implications of these findings, in vitro phagocytosis assays were performed. Azacitidine and venetoclax significantly enhanced AO-176-mediated phagocytosis of AML cells compared to any of the agents alone. Moreover, when AO-176 was combined with azacitidine in direct tumor cell killing assays, enhanced activity was observed in a subset of AML cell lines. In conclusion, AO-176 combined with either azacitidine or venetoclax, resulted in significant enhancement of phagocytic AML cell clearance in vitro which also correlated with the ability of these agents to induce DAMPs. In vivo treatment with AO-176 in combination with these agents is in progress. AO-176 is being evaluated in phase 1 clinical trials for the treatment of patients with solid tumors (NCT03834948) and multiple myeloma (NCT04445701). Disclosures Donio: Arch Oncology: Current Employment, Current equity holder in private company. Wilson:Arch Oncology: Current Employment, Current equity holder in private company. Darwech:Arch Oncology: Current Employment, Current equity holder in private company. Andrejeva:Arch Oncology: Current Employment, Current equity holder in private company. Capoccia:Arch Oncology: Current Employment, Current equity holder in private company. Puro:Arch Oncology: Current Employment, Current equity holder in private company. Kashyap:Arch Oncology: Current Employment, Current equity holder in private company. Pereira:Arch Oncology: Current Employment, Current equity holder in private company.