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Abstract Neuromyelitis optica spectrum disorders (NMOSD) comprise autoimmune diseases imposing substantial disability. We compared an NMOSD-targeted disability assessment of mobility, vision, and self-care domains (individually and composite) with the multiple sclerosis-targeted Expanded Disability Status Scale (EDSS) to assess NMOSD disease burden. An overall cohort (n = 505) and a subset of these patients with an enriched dataset (n = 198) were analyzed from the CIRCLES longitudinal, observational database of patients with AQP4-IgG–seropositive or –seronegative NMOSD in North America. Multinomial modeling was used to identify temporal correlates of disability improvement, stability, and worsening. Prior on-study relapse correlated with worsening mobility (OR, 3.08; 95% CI: 1.61–5.90), vision (OR, 3.99; 95% CI: 2.03–7.86), self-care disability (OR, 1.90; 95% CI: 1.07–3.38), and mean composite index disability (OR, 4.20; 95% CI: 1.71–10.34). Higher vision disability was associated with Black race, shorter time on-study, and AQP4-IgG–seropositive status in patients ≥ 18 years (p

Abstract Background Point-of-care ultrasound (POCUS) has emerged as an essential bedside tool for clinicians, but lack of access to ultrasound equipment has been a top barrier to POCUS use. Recently, several handheld ultrasound devices (“handhelds”) have become available, and clinicians are seeking data to guide purchasing decisions. Few comparative studies of different handhelds have been done. We conducted a cross-sectional study comparing 6 handhelds readily available in the United States (Butterfly iQ + ™ by Butterfly Network Inc.; Clarius™ by Clarius Mobile Health; Kosmos™ by EchoNous; TE Air™ by Mindray; Vscan Air™ SL and CL by General Electric; and Lumify™ by Philips Healthcare). A multi-specialty group of physician POCUS experts (n = 35) acquired three standard ultrasound views (abdominal right upper quadrant, cardiac apical 4-chamber, and superficial neck and lung views) in random order on the same standardized patients and rated the image quality. Afterward, a final survey of the overall ease of use, image quality, and satisfaction of each handheld was completed. Results Thirty-five POCUS experts specializing in internal medicine/hospital medicine, critical care, emergency medicine, and nephrology acquired and rated right upper quadrant, apical 4-chamber, and superficial neck and lung views with 6 different handhelds. For image quality, the highest-rated handhelds were Vscan Air™ for the right upper quadrant view, Mindray TE Air™ for the cardiac apical 4-chamber view, and Lumify™ for superficial views of the neck and lung. Overall satisfaction with image quality was highest with Vscan Air™, Lumify™, and Mindray, while overall satisfaction with ease of use was highest with Vscan Air™. The 5 most desirable characteristics of handhelds were image quality, ease of use, portability, probe size, and battery life. Ultimately, all 6 handhelds had notable advantages and disadvantages, with no single device having all desired qualities or features. Conclusions The overall satisfaction with image quality was rated highest with Vscan Air™, Lumify™, and Mindray TE Air™when acquiring right upper quadrant, apical 4-chamber, and superficial neck and lung views. No single handheld was perceived to be superior in image quality for all views. Vscan Air™ was rated highest for overall ease of use and was the most preferred handheld for purchase by POCUS experts.

Abstract Integrated crop‐livestock systems (ICLS) bring diversity to agricultural systems, enhancing soil ecosystem services, food production, and environmental sustainability. Resource utilization efficiency practices under semiarid ecoregions include dual systems that grow wheat (Triticum aestivum L.) for both grain and grazing (G) and recently complementary to wheat dual systems, cover crops (CC) for feeding both the soil and cattle during the fallow period. The latter continues to generate interest and there is a paucity of information on associated biochemical cycles. The objective was to evaluate the impact of CC and grazing thereof on soil microbiota structure, diversity, proliferation, and nutrient cycling. Introducing CC to no‐till (NT [NTC]) and grazing CC (NTCG [ICLS]), increased total PLFA biomass (TPB) for ungrazed CC (NTC) by 12%, and grazed CC (NTCG [ICLS]) by 20%; total bacteria biomass (TBB) by 10% for NTC and 24% for NTCG; total fungal biomass (TFB) by 9% for NTC and 21% for NTCG. The CC significantly increased Gram (−) bacteria biomass by 17% and 34% for NTC and NTCG, respectively; the CC significantly increased Gram (+) bacteria biomass by 6% and 12% for NTC and NTCG, respectively; and the CC significantly increased arbuscular mycorrhizal fungi by 55% and 89% for NTC and NTCG respectively, compared to NT fallow practice. Significant correlations were observed for NO3−–N, NH4+–N, water‐extractable organic nitrogen, total nitrogen, and water‐extractable organic carbon with TPB, TBB, and TFB using Haney soil health methods. Based on the measured parameters, the soil health status decreased in the order NTCG > NTC > NT > CT, where NT is the no‐till, C is the cover crop, G is the grazing, and CT is the conventional‐till. Grazing CC enhanced soil bacterial biomass over CC in solitude.

Sustainable crop production requires nutrient inputs to replace what was removed or lost. Replenishing nutrients using fertilizers and animal wastes is one of the most common input practices. While there is considerable research evaluating the efficacy of fertilizer and manure-based inputs for crop performance, nutrient use efficiency, and the subsequent impact to the environment, the influence of these inputs on soil health properties is less understood. The goals of this review are to summarize the state of literature on the influence of chemical fertilizer and manure-based inputs on soil health and to identify research gaps to refine efforts toward improved soil health, sustainability, and soil carbon storage. Conclusions suggest that chemical fertilizer inputs can increase soil organic matter and soil health provided that additions are not beyond what a plant can utilize. Manure-based inputs contribute to soil organic carbon and associated metrics; however, direct comparison of products is difficult as they are generally applied on a nutrient basis rather than carbon basis, which differentially affects their influence on soil health. Overall, nutrient-containing inputs show beneficial influences on soil health metrics; however, more focused research into their influence on soil health is needed.

Abstract Introduction Point-of-care ultrasound (POCUS) can assist rheumatologists in monitoring disease activity, establishing diagnoses, and guiding procedural interventions. POCUS use has been increasing, but little is known about current use and barriers among rheumatologists. The purpose of this study was to characterize current POCUS use, training needs, and barriers to use among rheumatologists in practice. Methods A prospective observational study of all Veterans Affairs (VA) medical centers was conducted using a web-based survey sent to all chiefs of staff and rheumatology chiefs about current POCUS use, training needs, barriers, and policies. Results All chiefs of staff (n = 130) and rheumatology chiefs at VA medical centers (n = 95) were surveyed with 100% and 84% response rates, respectively. The most common diagnostic POCUS applications were evaluation of synovitis, joint effusion, tendinopathies, bursitis, and rotator cuff. The most common procedural applications were arthrocentesis and joint, bursa, and tendon injection. Most rheumatology chiefs (69%) expressed interest in training for their group. The most common barriers to POCUS use were lack of trained providers (68%), funding for training (54%), training opportunities (38%), funding for travel (38%), and ultrasound equipment (31%). Lack of POCUS infrastructure was common, and few facilities had POCUS policies (20%), image archiving (25%), or quality assurance processes (6%). Conclusion Currently, half of rheumatology groups use diagnostic and procedural ultrasound applications. Most rheumatology groups desire training, and lack of training and equipment were the most common barriers to ultrasound use. Deliberate investment is needed in ultrasound training and infrastructure for systematic adoption of POCUS in rheumatology. Graphical Abstract available for this article. Trial Registration NCT03296280. Graphical Abstract

Abstract Durable host plant resistance (HPR) to insect pests is critical for sustainable agriculture. Natural variation exists for aphid HPR in sorghum (Sorghum bicolor), but the genetic architecture and phenotype have not been clarified and characterized for most sources. In order to assess the current threat of a sorghum aphid (Melanaphis sorghi) biotype shift, we characterized the phenotype of Resistance to Melanaphis sorghi 1 (RMES1) and additional HPR architecture in globally admixed populations selected under severe sorghum aphid infestation in Haiti. We found RMES1 reduces sorghum aphid fecundity but not bird cherry‐oat aphid (Rhopalosiphum padi) fecundity, suggesting a discriminant HPR response typical of gene‐for‐gene interaction. A second resistant gene, Resistance to Melanaphis sorghi 2 (RMES2), was more frequent than RMES1 resistant alleles in landraces and historic breeding lines. RMES2 contributes early and mid‐season aphid resistance in a segregating F2 population; however, RMES1 was only significant with mid‐season fitness. In a fixed population with high sorghum aphid resistance, RMES1 and RMES2 were selected for demonstrating a lack of severe antagonistic pleiotropy. Associations with resistance colocated with cyanogenic glucoside biosynthesis genes support additional HPR sources. Globally, therefore, an HPR source vulnerable to biotype shift via selection pressure (RMES1) is bolstered by a second common source of resistance in breeding programs (RMES2), which may be staving off a biotype shift and is critical for sustainable sorghum production.

Concentrations of specific secondary metabolites can be higher in organically grown crops. This may be linked to organic nitrogen (N) nutrition that provides a gradual supply of N to crops over the growing season. This study examined whether organic N nutrition influenced the concentration of glucosinolates in broccoli crops. Nitrogen release patterns were determined from three synthetic (Rustica, 12% N; calcium nitrate, 15.5% N; urea, 46% N) and two organic fertilizers in an incubation experiment. Broccoli seedlings were then grown in two N dose response pot trials with different N source or application timing treatments to investigate growth and glucosinolate responses. Synthetic fertilizers released 84 to 89% of total N after 28 days, while chicken manure pellets and composted cow manure had only released 52% and 13% of total N, respectively, after 91 days. Broccoli yield and N content were generally higher in synthetic fertilizer treatments. Glucosinolate concentrations were generally higher in the synthetic fertilizer treatments, and only sinigrin and glucoiberin concentrations in the 800 kg ha−1 N application rate of organic fertilizer matched those in the corresponding synthetic fertilizer treatment. Broccoli head weight was reduced when N was applied fortnightly compared to basal and weekly N applications, but glucosinolate concentrations were not significantly different. Overall, there was no evidence that organic (chicken manure) N nutrition, or the rate of N supply to broccoli plants, affect glucosinolate concentrations.

Executive summary Microbes are all pervasive in their distribution and influence on the functioning and well‐being of humans, life in general and the planet. Microbially‐based technologies contribute hugely to the supply of important goods and services we depend upon, such as the provision of food, medicines and clean water. They also offer mechanisms and strategies to mitigate and solve a wide range of problems and crises facing humanity at all levels, including those encapsulated in the sustainable development goals (SDGs) formulated by the United Nations. For example, microbial technologies can contribute in multiple ways to decarbonisation and hence confronting global warming, provide sanitation and clean water to the billions of people lacking them, improve soil fertility and hence food production and develop vaccines and other medicines to reduce and in some cases eliminate deadly infections. They are the foundation of biotechnology, an increasingly important and growing business sector and source of employment, and the centre of the bioeconomy, Green Deal, etc. But, because microbes are largely invisible, they are not familiar to most people, so opportunities they offer to effectively prevent and solve problems are often missed by decision‐makers, with the negative consequences this entrains. To correct this lack of vital knowledge, the International Microbiology Literacy Initiative–the IMiLI–is recruiting from the global microbiology community and making freely available, teaching resources for a curriculum in societally relevant microbiology that can be used at all levels of learning. Its goal is the development of a society that is literate in relevant microbiology and, as a consequence, able to take full advantage of the potential of microbes and minimise the consequences of their negative activities. In addition to teaching about microbes, almost every lesson discusses the influence they have on sustainability and the SDGs and their ability to solve pressing problems of societal inequalities. The curriculum thus teaches about sustainability, societal needs and global citizenship. The lessons also reveal the impacts microbes and their activities have on our daily lives at the personal, family, community, national and global levels and their relevance for decisions at all levels. And, because effective, evidence‐based decisions require not only relevant information but also critical and systems thinking, the resources also teach about these key generic aspects of deliberation. The IMiLI teaching resources are learner‐centric, not academic microbiology‐centric and deal with the microbiology of everyday issues. These span topics as diverse as owning and caring for a companion animal, the vast range of everyday foods that are produced via microbial processes, impressive geological formations created by microbes, childhood illnesses and how they are managed and how to reduce waste and pollution. They also leverage the exceptional excitement of exploration and discovery that typifies much progress in microbiology to capture the interest, inspire and motivate educators and learners alike. The IMiLI is establishing Regional Centres to translate the teaching resources into regional languages and adapt them to regional cultures, and to promote their use and assist educators employing them. Two of these are now operational. The Regional Centres constitute the interface between resource creators and educators–learners. As such, they will collect and analyse feedback from the end‐users and transmit this to the resource creators so that teaching materials can be improved and refined, and new resources added in response to demand: educators and learners will thereby be directly involved in evolution of the teaching resources. The interactions between educators–learners and resource creators mediated by the Regional Centres will establish dynamic and synergistic relationships–a global societally relevant microbiology education ecosystem–in which creators also become learners, teaching resources are optimised and all players/stakeholders are empowered and their motivation increased. The IMiLI concept thus embraces the principle of teaching societally relevant microbiology embedded in the wider context of societal, biosphere and planetary needs, inequalities, the range of crises that confront us and the need for improved decisioning, which should ultimately lead to better citizenship and a humanity that is more sustainable and resilient. Abstract The biosphere of planet Earth is a microbial world: a vast reactor of countless microbially driven chemical transformations and energy transfers that push and pull many planetary geochemical processes, including the cycling of the elements of life, mitigate or amplify climate change (e.g., Nature Reviews Microbiology, 2019, 17, 569) and impact the well‐being and activities of all organisms, including humans. Microbes are both our ancestors and creators of the planetary chemistry that allowed us to evolve (e.g., Life's engines: How microbes made earth habitable, 2023). To understand how the biosphere functions, how humans can influence its development and live more sustainably with the other organisms sharing it, we need to understand the microbes. In a recent editorial (Environmental Microbiology, 2019, 21, 1513), we advocated for improved microbiology literacy in society. Our concept of microbiology literacy is not based on knowledge of the academic subject of microbiology, with its multitude of component topics, plus the growing number of additional topics from other disciplines that become vitally important elements of current microbiology. Rather it is focused on microbial activities that impact us–individuals/communities/nations/the human world–and the biosphere and that are key to reaching informed decisions on a multitude of issues that regularly confront us, ranging from personal issues to crises of global importance. In other words, it is knowledge and understanding essential for adulthood and the transition to it, knowledge and understanding that must be acquired early in life in school. The 2019 Editorial marked the launch of the International Microbiology Literacy Initiative, the IMiLI. Here, we present our concept of how microbiology literacy may be achieved and the rationale underpinning it; the type of teaching resources being created to realise the concept and the framing of microbial activities treated in these resources in the context of sustainability, societal needs and responsibilities and decision‐making; and the key role of Regional Centres that will translate the teaching resources into local languages, adapt them according to local cultural needs, interface with regional educators and develop and serve as hubs of microbiology literacy education networks. The topics featuring in teaching resources are learner‐centric and have been selected for their inherent relevance, interest and ability to excite and engage. Importantly, the resources coherently integrate and emphasise the overarching issues of sustainability, stewardship and critical thinking and the pervasive interdependencies of processes. More broadly, the concept emphasises how the multifarious applications of microbial activities can be leveraged to promote human/animal, plant, environmental and planetary health, improve social equity, alleviate humanitarian deficits and causes of conflicts among peoples and increase understanding between peoples (Microbial Biotechnology, 2023, 16(6), 1091–1111). Importantly, although the primary target of the freely available (CC BY‐NC 4.0) IMiLI teaching resources is schoolchildren and their educators, they and the teaching philosophy are intended for all ages, abilities and cultural spectra of learners worldwide: in university education, lifelong learning, curiosity‐driven, web‐based knowledge acquisition and public outreach. The IMiLI teaching resources aim to promote development of a global microbiology education ecosystem that democratises microbiology knowledge.

The development of subsurface microstructural alterations known as dark etching regions (DERs) and white etching bands (WEBs) in rolling element bearings due to rolling contact fatigue have been investigated for the past eight decades, focusing on their initiation and formation mechanisms. They have only recently been shown to be driven by repetitive cycles of energy build-up due to micro-plastic deformation and energy release through recrystallization and recovery, which results in the formation of equiaxed and elongated ferrite grains, as well as lenticular carbides. These features develop within the bearing subsurface from DER to WEBs during bearing operation at moderate to high loads, but little evidence has been presented in the literature to understand links between DER and WEBs and the nucleation and growth of subsurface cracks. This investigation examines WEBs, including low angle bands (LABs) and high angle bands (HABs), in detail especially focusing on their late stages to understand such links. A number of techniques, including optical microscopy, scanning electron microscopy (SEM) and Energy-dispersive X-ray spectroscopy (EDX) have been used to examine the features involved. Analysis on WEBs obtained through serial sectioning has revealed that voids initiating at the interface between lenticular carbides and equiaxed ferrite grain bands within WEBs have led to crack formation which can subsequently propagate to bearing surfaces. Interactions between WEBs and non-metallic inclusions (NMIs) are observed to lead to de-bonding of inclusions from their surrounding microstructure and void formation, which has also found to influence the integrity of the bearings at late stages. Alumina and Manganese sulphide (MnS) inclusions are the mostly observed NMIs that de-bond and develop microcracks when interacting with WEBs. These findings thus provide important insights into the link between inclusions and crack initiation and represent a further step towards a fundamental understanding of the rolling contact fatigue process.

This study examined the effects of sustained, isometric forearm flexion tasks anchored to ratings of perceived exertion of 2 (RPE2FT) and 8 (RPE8FT) on the patterns of fatigue-induced changes in torque and neuromuscular responses, time to task failure (TTF), performance fatigability (% decline in maximal voluntary isometric contraction [MVIC]), and perceived factors that contributed to task termination. Twelve men (mean ± SD: age = 20.9 ± 2.2 yrs) performed MVICs before and after the tasks and completed post-test questionnaires (PTQ). Data were analyzed using polynomial regression analyses, dependent t-tests, Spearman’s rank order correlations, and Wilcoxon signed rank tests. The RPE8FT had greater (p < 0.001) performance fatigability than the RPE2FT, despite no difference (p > 0.05) in TTF. During both tasks, there were significant (p ≤ 0.05) composite linear decreases for torque, electromyographic amplitude, and neuromuscular efficiency, and substantial individual variability in the neuromuscular responses. There were no significant (p > 0.05) associations among the perceived factors and TTF or performance fatigability. Thus, there were RPE-specific differences in performance fatigability, but not TTF or the composite patterns of changes in torque and neuromuscular responses. In addition, in most cases, the individual neuromuscular, but not torque, patterns of responses were RPE-specific. Furthermore, the perceived factors assessed by the PTQ were not related to TTF or performance fatigability.

Abstract Global population growth makes it necessary to increase agricultural production yields. However, climate change impacts and diseases caused by plant pathogens are challenging modern agriculture. Therefore, it is necessary to look for alternatives to the excessive use of chemical fertilizers and pesticides. The plant microbiota plays an essential role in plant nutrition and health, and offers enormous potential to meet future challenges of agriculture. In this context, here we characterized the antifungal properties of the rhizosphere bacterium Pantoea agglomerans 9Rz4, which is active against a broad spectrum of plant pathogenic fungi. Chemical analyses revealed that strain 9Rz4 produces the antifungal herbicolin A and its biosynthetic gene cluster was identified and characterized. We found that the only acyl‐homoserine lactone‐based quorum sensing system of 9Rz4 modulates herbicolin A gene cluster expression. No role of plasmid carriage in the production of herbicolin A was observed. Plant assays revealed that herbicolin A biosynthesis does not affect the root colonization ability of P. agglomerans 9Rz4. Current legislative restrictions are aimed at reducing the use of chemical pesticides in agriculture, and the results derived from this study may lay the foundations for the development of novel biopesticides from rhizosphere microorganisms.

Abstract Background Frailty and dementia have a bidirectional relationship. However, frailty is rarely reported in clinical trials for dementia and mild cognitive impairment (MCI) which limits assessment of trial applicability. This study aimed to use a frailty index (FI)—a cumulative deficit model of frailty—to measure frailty using individual participant data (IPD) from clinical trials for MCI and dementia. Moreover, the study aimed to quantify the prevalence of frailty and its association with serious adverse events (SAEs) and trial attrition. Methods We analysed IPD from dementia (n = 1) and MCI (n = 2) trials. An FI comprising physical deficits was created for each trial using baseline IPD. Poisson and logistic regression were used to examine associations with SAEs and attrition, respectively. Estimates were pooled in random effects meta-analysis. Analyses were repeated using an FI incorporating cognitive as well as physical deficits, and results compared. Results Frailty could be estimated in all trial participants. The mean physical FI was 0.14 (SD 0.06) and 0.14 (SD 0.06) in the MCI trials and 0.24 (SD 0.08) in the dementia trial. Frailty prevalence (FI > 0.24) was 6.9%/7.6% in MCI trials and 48.6% in the dementia trial. After including cognitive deficits, the prevalence was similar in MCI (6.1% and 6.7%) but higher in dementia (75.4%). The 99th percentile of FI (0.31 and 0.30 in MCI, 0.44 in dementia) was lower than in most general population studies. Frailty was associated with SAEs: physical FI IRR = 1.60 [1.40, 1.82]; physical/cognitive FI IRR = 1.64 [1.42, 1.88]. In a meta-analysis of all three trials, the estimated association between frailty and trial attrition included the null (physical FI OR = 1.17 [0.92, 1.48]; physical/cognitive FI OR = 1.16 [0.92, 1.46]), although higher frailty index values were associated with attrition in the dementia trial. Conclusion Measuring frailty from baseline IPD in dementia and MCI trials is feasible. Those living with more severe frailty may be under-represented. Frailty is associated with SAEs. Including only physical deficits may underestimate frailty in dementia. Frailty can and should be measured in future and existing trials for dementia and MCI, and efforts should be made to facilitate inclusion of people living with frailty.

Haemonchus contortus is a parasitic haematophagous nematode that primarily affects small ruminants and causes significant economic loss to the global livestock industry. Treatment of haemonchosis typically relies on broad-spectrum anthelmintics, resistance to which is an important cause of treatment failure. Resistance to levamisole remains less widespread than to other major anthelmintic classes, prompting the need for more effective and accurate surveillance to maintain its efficacy. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) is a recently developed diagnostic method that facilitates multiplex target detection with single nucleotide polymorphism (SNP) specificity and portable onsite testing. In this study, we designed a new LEC-LAMP assay and applied it to detect the levamisole resistance marker S168T in H. contortus. We explored multiplexing probes for both the resistant S168T and the susceptible S168 alleles in a single-tube assay. We then included a generic probe to detect the acr-8 gene in the multiplex assay, which could facilitate the quantification of both resistance markers and overall genetic material from H. contortus in a single step. Our results showed promising application of these technologies, demonstrating a proof-of-concept assay which is amenable to detection of resistance alleles within the parasite population, with the potential for multiplex detection, and point-of-care application enabled by lateral flow end-point detection. However, further optimisation and validation is necessary.

Pigmented rice, especially black rice, is gaining popularity as it is rich in antioxidants such as anthocyanins and γ-oryzanol. At present, knowledge about temporal control of biosynthesis and accumulation of antioxidants during grain development is limited. To address this, the accumulation patterns of anthocyanins and γ-oryzanol were assessed in two distinct black rice genotypes over the course of grain development, and the expression of known regulatory genes for anthocyanin biosynthesis was examined. The results indicated that total γ-oryzanol content increased continuously throughout grain development, while total anthocyanins peaked at dough stage (15 to 21 days after flowering) followed by a decline until grain maturity in both genotypes. However, the rate of decrease in anthocyanin content differed between genotypes, and a more prominent decline in cyanidin 3-O-glucoside (C3G) relative to peonidin 3-O-glucoside (P3G) was observed for both. Anthocyanin content was closely linked with the expression of key regulatory genes in the MBW (MYB-bHLH-WD40) complex. This improved knowledge of the genotype-specific biosynthesis (anthocyanins only) and accumulation patterns of anthocyanins and γ-oryzanol can inform subsequent research efforts to increase concentrations of these key antioxidants in black rice grains.

Arthropod-borne viruses are major causes of human and animal disease, especially in endemic low- and middle-income countries. Mosquito-borne pathogen surveillance is essential for risk assessment and vector control responses. Sentinel chicken serosurveillance (antibody testing) and mosquito pool screening (by RT-qPCR or virus isolation) are currently used to monitor arbovirus transmission, however substantial time lags of seroconversion and/or laborious mosquito identification and RNA extraction steps sacrifice their early warning value. As a consequence, timely vector control responses are compromised. Here, we report on development of a rapid arbovirus detection system whereby adding sucrose to reagents of loop-mediated isothermal amplification with displaced probes (DP-LAMP) elicits infectious mosquitoes to feed directly upon the reagent mix and expectorate viruses into the reagents during feeding. We demonstrate that RNA from pathogenic arboviruses (West Nile and Dengue viruses) transmitted in the infectious mosquito saliva was detectable rapidly (within 45 minutes) without RNA extraction. Sucrose stabilized viral RNA at field temperatures for at least 48 hours, important for transition of this system to practical use. After thermal treatment, the DP-LAMP could be reliably visualized by a simple optical image sensor to distinguish between positive and negative samples based on fluorescence intensity. Field application of this technology could fundamentally change conventional arbovirus surveillance methods by eliminating laborious RNA extraction steps, permitting arbovirus monitoring from additional sites, and substantially reducing time needed to detect circulating pathogens.

There is debate about whether the honey from Leptospermum scoparium nectar in New Zealand is substantially different to honey from Leptospermum spp nectar from Australia; many Leptospermum honeys have high amounts of the antibacterial compound methylglyoxal (MGO) but are known to differ in their amounts of other metabolites. Retail honey samples labelled as ‘mānuka’, manuka, or ‘tea tree’ honey sourced from New Zealand (n = 34) and Australia (n = 35), were measured using laser assisted-rapid evaporative ionisation mass spectrometry (REIMS) in positive and negative ionisation modes, with 1637 and 1744 molecular features detected, respectively. Country of origin was strongly reflected in the REIMS fingerprints irrespective of similarity of methylglyoxal, with >50% of detected features differing between New Zealand and Australian-sourced honey. Combined REIMS features in negative ionisation mode correlated strongly with current molecular markers of mānuka honey quality (r2 > 0.9). Leptospermum honey from New Zealand and Australia have distinct molecular fingerprints, potentially due to evolutionary and genomic differences between the predominant Leptospermum species, bees, environment and honey processing.

Abstract Background The statistical models developed for meta-analysis of diagnostic test accuracy studies require specialised knowledge to implement. This is especially true since recent guidelines, such as those in Version 2 of the Cochrane Handbook of Systematic Reviews of Diagnostic Test Accuracy, advocate more sophisticated methods than previously. This paper describes a web-based application - MetaBayesDTA - that makes many advanced analysis methods in this area more accessible. Results We created the app using R, the Shiny package and Stan. It allows for a broad array of analyses based on the bivariate model including extensions for subgroup analysis, meta-regression and comparative test accuracy evaluation. It also conducts analyses not assuming a perfect reference standard, including allowing for the use of different reference tests. Conclusions Due to its user-friendliness and broad array of features, MetaBayesDTA should appeal to researchers with varying levels of expertise. We anticipate that the application will encourage higher levels of uptake of more advanced methods, which ultimately should improve the quality of test accuracy reviews.

Abstract Environmental water is considered one of the main vehicles for the transmission of antimicrobial resistance (AMR), posing an increasing threat to humans and animals health. Continuous efforts are being made to eliminate AMR; however, the detection of AMR pathogens from water samples often requires at least one culture step, which is time‐consuming and can limit sensitivity. In this study, we employed comparative genomics to identify the prevalence of AMR genes within among: Escherichia coli, Klebsiella, Salmonella enterica and Acinetobacter, using publicly available genomes. The mcr‐1, blaKPC (KPC‐1 to KPC‐4 alleles), blaOXA‐48, blaOXA‐23 and blaVIM (VIM‐1 and VIM‐2 alleles) genes are of great medical and veterinary significance, thus were selected as targets for the development of isothermal loop‐mediated amplification (LAMP) detection assays. We also developed a rapid and sensitive sample preparation method for an integrated culture‐independent LAMP‐based detection from water samples. The developed assays successfully detected the five AMR gene markers from pond water within 1 h and were 100% sensitive and specific with a detection limit of 0.0625 μg/mL and 10 cfu/mL for genomic DNA and spiked bacterial cells, respectively. The integrated detection can be easily implemented in resource‐limited areas to enhance One Health AMR surveillances and improve diagnostics.

Abstract Manufacturing autologous chimeric antigen receptor (CAR) T cell therapeutics is complex, and many patients experience treatment delays or cannot be treated at all. Although current allogeneic CAR products have the potential to overcome manufacturing bottlenecks, they are subject to immune rejection and failure to persist in the host, and thus do not provide the same level of efficacy as their autologous counterparts. Here, we aimed to develop universal allogeneic CAR T cells that evade the immune system and produce a durable response. We generated human hypoimmune (HIP) T cells with disrupted B2M, CIITA, and TRAC genes using CRISPR-Cas9 editing. In addition, CD47 and anti-CD19 CAR were expressed using lentiviral transduction. These allogeneic HIP CD19 CAR T cells were compared to allogeneic CD19 CAR T cells that only expressed the anti-CD19 CAR (allo CAR T). In vitro assays for cancer killing and exhaustion revealed no differences between allo CAR T and HIP CAR T cells, confirming that the HIP edits did not negatively affect T cell performance. Clearance of CD19+ tumors by HIP CAR T cells in immunodeficient NSG mice was comparable to that of allo CAR T cells. In fully immunocompetent humanized mice, HIP CAR T cells significantly outperformed allo CAR T cells, showed improved persistence and expansion, and provided lasting cancer clearance. Furthermore, CD47-targeting safety strategies reliably and specifically eliminated HIP CAR T cells. These findings suggest that universal allogeneic HIP CAR T cell-based therapeutics might overcome the limitations associated with poor persistence of allogeneic CAR T cells and exert durable anti-tumor responses.

Accumulation of phytochemicals in vegetative tissue under nitrogen (N) stress as an adaptive strategy has been investigated in various crops, but the effect of applied N on grain phytochemicals is poorly understood. This study investigated the effect of applied N on the biosynthesis and accumulation of rice (Oryza sativa L.) grain anthocyanin and γ-oryzanol under different ultraviolet-B (UV-B) conditions in a controlled pot trial using two distinct black rice genotypes. The response of grain anthocyanin and γ-oryzanol content to applied N was genotype-dependent but was not altered by UV-B conditions. Applied N increased grain anthocyanin and decreased γ-oryzanol content in genotype SCU212 but had no significant effect in genotype SCU254. The expression of the OsKala3 regulatory gene was significantly upregulated in response to applied N in SCU212, while the expressions of OsKala4 and OsTTG1 were unchanged. The expression of all three regulatory genes was not significantly affected in SCU254 with applied N. Key anthocyanin biosynthesis genes were upregulated in grain by N application, which indicates that the common increase in anthocyanin in vegetative tissues under N deprivation does not hold true for reproductive tissues. Hence, any future approach to target higher content of these key phytochemicals in grains should be genotype-focused.

ObjectiveTo determine the impact of thyroid eye disease (TED) on patients in various stages of the disease.BackgroundTED is a debilitating and potentially sight-threatening inflammatory autoimmune disease that is frequently misdiagnosed. Challenging quality-of-life (QoL) issues can persist long after the active phase of disease has subsided.MethodsA 62-question survey was designed as a hypothesis-generating instrument to identify key issues confronting patients ≥18 years old with physician-diagnosed TED. Questions focused primarily on physical and emotional status, and QoL experiences in the 2 months prior to the survey. Data for individual questions are presented as summary statistics. Correlations between questions were determined using χ2 analyses.ResultsThe 443 respondents were 18 to >80 years old; >90% female, and >80% from the United States. Time since TED diagnosis ranged from 10 years. Participants provided >500 free-form responses describing experiences of living with TED. Physical signs/symptoms were experienced by 307/443 (69%) patients. Of those responding to the QoL questions (N = 394), 53 (13%) reported symptoms improving, 73 (19%) reported symptoms worsening, and 255 (65%) reported no change in the 2 months prior to the survey. The most bothersome signs/symptoms were dry/gritty eyes, light sensitivity, bulging eyes, and pressure or pain behind the eyes. Respondents 5 symptoms, had been diagnosed within the last 5 years, or were

Background Treatment burden is the workload of healthcare for people with long-term conditions and the impact on wellbeing. A validated measure of treatment burden after stroke is needed. We aim to adapt a patient-reported measure (PRM) of treatment burden in multimorbidity, PETS (Patient Experience with Treatment and Self-Management version 2.0), to create a stroke-specific measure, PETS-stroke. We aim to examine content validity, construct validity, reliability and feasibility in a stroke survivor population. Methods 1) Adaptation of 60-item PETS to PETS-stroke using a taxonomy of treatment burden. 2) Content validity testing through cognitive interviews that will explore the importance, relevance and clarity of each item. 3) Evaluation of scale psychometric properties through analysis of data from stroke survivors recruited via postal survey (n=340). Factor structure will be tested with confirmatory factor analysis and Cronbach’s alpha will be used to index internal consistency. Construct validity will be tested against: The Stroke Southampton Self-Management Questionnaire; The Satisfaction with Stroke Care Measure; and The Shortened Stroke Impact Scale. We will explore known-groups validity by exploring the association between treatment burden, socioeconomic deprivation and multimorbidity. Test-retest reliability will be examined via re-administration after 2 weeks. Acceptability and feasibility of use will be explored via missing data rates and telephone interviews with 30 participants. Conclusions We aim to create a validated PRM of treatment burden after stroke. PETS-stroke is designed for use as an outcome measure in clinical trials of stroke treatments and complex interventions to ascertain if treatments are workable for patients in the context of their everyday lives.

Abstract Background Regular participation in resistance exercise is known to have broad-ranging health benefits and for this reason is prominent in the current physical activity guidelines. Recovery after such exercise is important for several populations across the age range and nutritional strategies to enhance recovery and modulate post-exercise physiological processes are widely studied, yet effective strategies remain elusive. Vitamin K2 supplementation has emerged as a potential candidate, and the aim of the current study, therefore, is to test the hypothesis that vitamin K2 supplementation can accelerate recovery, via modulation of the underlying physiological processes, following a bout of resistance exercise in young and older adults. Methods The current study is a two-arm randomised controlled trial which will be conducted in 80 (40 young (≤40 years) and 40 older (≥65 years)) adults to compare post-exercise recovery in those supplemented with vitamin K2 or placebo for a 12-week period. The primary outcome is muscle strength with secondary outcomes including pain-free range of motion, functional abilities, surface electromyography (sEMG) and markers of inflammation and oxidative stress. Discussion Ethical approval has been granted by the College of Medical Veterinary and Life Sciences Ethical Committee at the University of Glasgow (Project No 200190189) and recruitment is ongoing. Study findings will be disseminated through a presentation at scientific conferences and in scientific journals. Trial registration ClinicialTrials.gov NCT04676958. Prospectively registered on 21 December 2020.

Abstract Background Animal migration is a widespread global adaptation by which individuals move in response to environmental conditions to reach more favorable conditions. For bats in temperate climates, migration and hibernation are often associated with each other when these bats must migrate to reach suitable overwintering sites. However, differences in movement across the geographical range of a species and the degree to which hibernation drives migratory behavior of bats in subtropical climates, where conditions may remain warm with available prey year-round, remains incomplete. Understanding the migratory strategies of subtropical bats during winter is of increasing importance as they are threatened by stressors such as disease and environmental change. Methods We evaluated migration patterns of tricolored bats (Perimyotis subflavus) in Florida, USA, through analysis of stable hydrogen isotope ratios of the fur. We inferred the summer geographic origins of the fur samples and estimated the minimum distance and likely direction traveled by hibernating individuals. We used linear models to examine whether hibernation region, colony size, and an individual’s sex affected the distance traveled. Results We sampled 111 bats hibernating at 40 sites and found that more than half (54.1%) of individuals were residents of the area in which they hibernated. We found that almost half of the sampled bats (43.2%) traveled from southern Florida to overwinter in North Florida. We also documented three individuals that traveled > 100 km from northerly origins, one of which had traveled an estimated minimum distance of 1382 km. We also found that tricolored bats moved farther to reach hibernacula in Northwest Florida and hibernacula with more populous colonies, with no difference in movement between sexes. Conclusions Our results indicate a pattern of northward autumnal movements of tricolored bats in the subtropical southeastern portion of their range. We suggest that bats are differentially constrained at the edge of their geographical range, resulting in movement contrary to what is expected. Even though we found that few (2.7%) bats moved into Florida from farther north, those migrants can potentially transfer the fungus that causes the deadly white-nose syndrome, which does not currently occur in the state. Our results support the characterization of tricolored bats as flexible partial migrators, with a rarely exercised capacity for long-distance movements.

Legume cover crops in temperate cropping systems can fix substantial amounts of nitrogen (N) and reduce N fertiliser requirements for subsequent crops. However, little is known about potential biological N2 fixation by summer cover crop legumes in the short summer fallow in Mediterranean-type cropping systems. Six legume species (balansa clover, barrel medic, mung bean, sunn hemp, lablab and cowpea) were grown for 8–9 weeks in the field in semi-arid southern Australia during the summer fallow, and in a glasshouse experiment, to estimate N2 fixation using the 15N natural abundance method. Cowpea, sunn hemp and lablab produced 1.2–3.0 t ha−1 biomass in the field while balansa clover and barrel medic produced < 1.0 t ha−1. The percent of N derived from the atmosphere (%Ndfa) in the field ranged from 39% in barrel medic to 73% in sunn hemp, but only 15% (balansa clover) to 33% (sunn hemp) in the glasshouse experiment, likely due to higher soil mineral N availability in the glasshouse study. Biological N2 fixation of cowpea and sunn hemp in the field was 46–55 kg N ha−1, while N2 fixation in lablab and mung bean was lower (around 26 kg N ha−1). The N2 fixation in cowpea and sunn hemp of around 50 kg N ha−1 with supplementary irrigation in the field trial likely represents the upper limit of N contributions in the field in typically hot, dry summer conditions in Mediterranean-type climates. Given that any increase in summer cover crop biomass will have implications for water balances and subsequent cash crop growth, maximising N benefits of legume cover crops will rely on increasing the %Ndfa through improved rhizobium strains or inoculation technologies. This study provides the first known estimates of biological N2 fixation by legume cover crops in the summer fallow period in cropping systems in Mediterranean-type environments, providing a benchmark for further studies.

Autosomal dominant polycystic kidney disease (ADPKD) is a complicated disease that involves numerous cell types. Here the authors used a multiomics approach consisting of single nucleus transcriptomes and epigenomes to redefine cell states in ADPKD and to dissect the cellular interactions and molecular mechanisms of ADPKD.

A decadal-scale field trial revealed 1.01 Mg of rhizodeposit and necromass C was stored in soil microaggregate and mineral fractions per Mg biochar-C applied. Microspectroscopic analyses visualize mechanisms for this elevated soil C storage ceiling.

Background Treatment burden is the workload of healthcare for people with long-term conditions and the impact on wellbeing. A validated measure of treatment burden after stroke is needed. We aim to adapt a patient-reported measure (PRM) of treatment burden in multimorbidity, PETS (Patient Experience with Treatment and Self-Management version 2.0), to create a stroke-specific measure, PETS-stroke. We aim to examine content validity, construct validity, reliability and feasibility in a stroke survivor population. Methods 1) Adaptation of 60-item PETS to PETS-stroke using a taxonomy of treatment burden. 2) Content validity testing through cognitive interviews that will explore the importance, relevance and clarity of each item. 3) Evaluation of scale psychometric properties through analysis of data from stroke survivors recruited via postal survey (n=340). Factor structure will be tested with confirmatory factor analysis and Cronbach’s alpha will be used to index internal consistency. Construct validity will be tested against: The Stroke Southampton Self-Management Questionnaire; The Satisfaction with Stroke Care Measure; and The Shortened Stroke Impact Scale. We will explore known-groups validity by exploring the association between treatment burden, socioeconomic deprivation and multimorbidity. Test-retest reliability will be examined via re-administration after 2 weeks. Acceptability and feasibility of use will be explored via missing data rates and telephone interviews with 30 participants. Conclusions We aim to create a validated PRM of treatment burden after stroke. PETS-stroke is designed for use as an outcome measure in clinical trials of stroke treatments and complex interventions to ascertain if treatments are workable for patients in the context of their everyday lives.

Background Hematologic toxicities, including coagulopathy, endothelial activation, and cytopenias, with CD19-targeted chimeric antigen receptor (CAR) T-cell therapies correlate with cytokine release syndrome (CRS) and neurotoxicity severity, but little is known about the extended toxicity profiles of CAR T-cells targeting alternative antigens. This report characterizes hematologic toxicities seen following CD22 CAR T-cells and their relationship to CRS and neurotoxicity.Methods We retrospectively characterized hematologic toxicities associated with CRS seen on a phase 1 study of anti-CD22 CAR T-cells for children and young adults with relapsed/refractory CD22+ hematologic malignancies. Additional analyses included correlation of hematologic toxicities with neurotoxicity and exploring effects of hemophagocytic lymphohistiocytosis-like toxicities (HLH) on bone marrow recovery and cytopenias. Coagulopathy was defined as evidence of bleeding or abnormal coagulation parameters. Hematologic toxicities were graded by Common Terminology Criteria for Adverse Events V.4.0.Results Across 53 patients receiving CD22 CAR T-cells who experienced CRS, 43 (81.1%) patients achieved complete remission. Eighteen (34.0%) patients experienced coagulopathy, of whom 16 had clinical manifestations of mild bleeding (typically mucosal bleeding) which generally subsided following CRS resolution. Three had manifestations of thrombotic microangiopathy. Patients with coagulopathy had higher peak ferritin, D-dimer, prothrombin time, international normalized ratio (INR), lactate dehydrogenase (LDH), tissue factor, prothrombin fragment F1+2 and soluble vascular cell adhesion molecule-1 (s-VCAM-1). Despite a relatively higher incidence of HLH-like toxicities and endothelial activation, overall neurotoxicity was generally less severe than reported with CD19 CAR T-cells, prompting additional analysis to explore CD22 expression in the central nervous system (CNS). Single-cell analysis revealed that in contrast to CD19 expression, CD22 is not on oligodendrocyte precursor cells or on neurovascular cells but is seen on mature oligodendrocytes. Lastly, among those attaining CR, grade 3–4 neutropenia and thrombocytopenia were seen in 65% of patients at D28.Conclusion With rising incidence of CD19 negative relapse, CD22 CAR T-cells are increasingly important for the treatment of B-cell malignancies. In characterizing hematologic toxicities on CD22 CAR T-cells, we demonstrate that despite endothelial activation, coagulopathy, and cytopenias, neurotoxicity was relatively mild and that CD22 and CD19 expression in the CNS differed, providing one potential hypothesis for divergent neurotoxicity profiles. Systematic characterization of on-target off-tumor toxicities of novel CAR T-cell constructs will be vital as new antigens are targeted.Trial registration number NCT02315612.

Saline agriculture, including the commercial cultivation of edible halophytes, is expanding rapidly to address the increase in salinised soil due to natural and man-made causes, the decline in availability of fresh water, the increased use of poor-quality water, and increasing food insecurity. Halophytes, as food crops, offer an innovative new opportunity in agriculture, many being highly nutritious and containing bioactive compounds. However, the commercial production of edible halophytes to meet emerging consumer demands faces several challenges. This review examines the market potential for the expansion of edible halophyte crops and the challenges of consumer acceptance and production capacity beyond wild harvest. In addition to beneficial compounds, halophytes are known to contain several anti-nutrient compounds, which can have negative health consequences. In edible halophytes, oxalates are of particular concern. However, research has shown that there are ways to mitigate the accumulation of oxalate through processing, agronomic practice, and genetic engineering. These approaches are presented as potential strategies that can be used in commercial farming systems to reduce the levels of this compound.

Abstract The Rolling Plains of Texas are historically semi‐arid with sporadic, high intensity storms followed by periods of long drought. Fallow management is a common practice intended to store soil water, but leaves the bare soil exposed to erosive forces that can diminish the soil productivity. Cover crops in no‐till (NT) agriculture have been proposed to increase soil health under environments with low precipitation as an alternative to fallow management. This study evaluated multiple treatments in a dryland cotton system including: (1) conventional tillage (CT); (2) NT; and NT with the following cover crops: (3) wheat; (4) Austrian winter pea; (5) crimson clover; (6) hairy vetch; and (7) mixed species cover. Soil samples were collected at 0, 3, and 6 weeks after cover crop termination and analyzed for soil organic carbon, total nitrogen, inorganic N, water‐extractable organic carbon, water‐extractable organic nitrogen, carbon mineralization, and phospholipid fatty acid analysis. For all parameters tested, there was no significant difference between CT and NT at any date or depth, so the addition of cover crops to NT cotton systems might be needed in order to enhance NT in regard to soil function. The multi‐species mixed treatment was predicted to perform the best out of the cover crop treatments due to its combined benefits from grasses and legumes. However, the single‐species Austrian winter pea treatment had 24% and 28% higher soil carbon and nitrogen than no‐till without a cover crop, and can be a useful alternative to fallow management under these dryland agriculture conditions.

Legume nodulation is the powerhouse of biological nitrogen fixation (BNF) where host-specific rhizobia dominate the nodule microbiome. However, other rhizobial or non-rhizobial inhabitants can also colonize legume nodules, and it is unclear how these bacteria interact, compete, or combinedly function in the nodule microbiome. Under such context, to test this hypothesis, we conducted 16S-rRNA based nodule microbiome sequencing to characterize microbial communities in two distinct sized nodules from field-grown peanuts inoculated with a commercial inoculum. We found that microbial communities diverged drastically in the two types of peanut nodules (big and small). Core microbial analysis revealed that the big nodules were inhabited by Bradyrhizobium, which dominated composition (>99%) throughout the plant life cycle. Surprisingly, we observed that in addition to Bradyrhizobium, the small nodules harbored a diverse set of bacteria (~31%) that were not present in big nodules. Notably, these initially less dominant bacteria gradually dominated in small nodules during the later plant growth phases, which suggested that native microbial communities competed with the commercial inoculum in the small nodules only. Conversely, negligible or no competition was observed in the big nodules. Based on the prediction of KEGG pathway analysis for N and P cycling genes and the presence of diverse genera in the small nodules, we foresee great potential of future studies of these microbial communities which may be crucial for peanut growth and development and/or protecting host plants from various biotic and abiotic stresses.

Private wells are susceptible to contamination from flooding and are exempt from the federal requirements of the Safe Drinking Water Act. Consequently, well users must manage (e.g., disinfect) and maintain (e.g., regularly test) their own wells to ensure safe drinking water. However, well user practices and perceptions of well water quality in the years following a natural disaster are poorly characterized. An online follow-up survey was administered in October 2020 to private well users who had previously experienced Hurricane Harvey in 2017. The survey was successfully sent to 436 participants, and 69 surveys were returned (15.8% return rate). The survey results indicate that well users who had previously experienced wellhead submersion or a positive bacteria test were more likely to implement well stewardship practices (testing and disinfection) and to report the feeling that their well water was safe. While the majority of well users believed that their water was safe (77.6%), there was a significant decrease in well water being used for drinking, cooking, and for their pets after Hurricane Harvey. Generally, these well users tend to maintain their wells at higher rates than those reported in other communities, but there continues to be a critical need to provide outreach regarding well maintenance practices, especially before natural disaster events occur.

ABSTRACT Cefotaximase-Munich (CTX-M) extended-spectrum beta-lactamase (ESBL) enzymes produced by Enterobacteriaceae confer resistance to clinically relevant third-generation cephalosporins. CTX-M group 1 variants, CTX-M-1 and CTX-M-15, are the leading ESBL-producing Enterobacteriaceae associated with animal and human infection, respectively, and are an increasing antimicrobial resistance (AMR) global health concern. The blaCTX-M-1 and blaCTX-M-15 genes encoding these variants have an approximate nucleotide sequence similarity of 98.7%, making effective differential diagnostic monitoring difficult. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) enables rapid real-time multiplex pathogen detection with single-base specificity and portable on-site testing. We have developed an internally controlled multiplex CTX-M-1/15 LEC-LAMP assay for the differential detection of blaCTX-M-1 and blaCTX-M-15. Assay analytical specificity was established using a panel of human, animal, and environmental Escherichia coli isolates positive for blaCTX-M-1 (n = 18), blaCTX-M-15 (n = 35), and other closely related blaCTX-Ms (n = 38) from Ireland, Germany, and Portugal, with analytical sensitivity determined using probit regression analysis. Animal fecal sample testing using the CTX-M-1/15 LEC-LAMP assay in combination with a rapid DNA extraction protocol was carried out on porcine fecal samples previously confirmed to be PCR-positive for E. coli blaCTX-M. Portable instrumentation was used to further analyze each fecal sample and demonstrate the on-site testing capabilities of the LEC-LAMP assay with the rapid DNA extraction protocol. The CTX-M-1/15 LEC-LAMP assay demonstrated complete analytical specificity for the differential detection of both variants with sensitive low-level detection of 8.5 and 9.8 copies per reaction for blaCTX-M-1 and blaCTX-M-15, respectively, and E. coli blaCTX-M-1 was identified in all blaCTX-M positive porcine fecal samples tested. IMPORTANCE CTX-M ESBL-producing E. coli is an increasing AMR public health issue with the transmission between animals and humans via zoonotic pathogens now a major area of interest. Accurate and timely identification of ESBL-expressing E. coli CTX-M variants is essential for disease monitoring, targeted antibiotic treatment and infection control. This study details the first report of portable diagnostics technology for the rapid differential detection of CTX-M AMR markers blaCTX-M-1 and blaCTX-M-15, facilitating improved identification and surveillance of these closely related variants. Further application of this portable internally controlled multiplex CTX-M-1/15 LEC-LAMP assay will provide new information on the transmission and prevalence of these CTX-M ESBL alleles. Furthermore, this transferable diagnostic technology can be applied to other new and emerging relevant AMR markers of interest providing more efficient and specific portable pathogen detection for improved epidemiological surveillance.

Acetylation of lysine residues is an important and common post-translational regulatory mechanism occurring on thousands of non-histone proteins. Lysine deacetylases (KDACs or HDACs) are a family of enzymes responsible for removing acetylation. To identify the biological mechanisms regulated by individual KDACs, we created HT1080 cell lines containing chromosomal point mutations, which endogenously express either KDAC6 or KDAC8 having single inactivated catalytic domain. Engineered HT1080 cells expressing inactive KDA6 or KDAC8 domains remained viable and exhibited enhanced acetylation on known substrate proteins. RNA-seq analysis revealed that many changes in gene expression were observed when KDACs were inactivated, and that these gene sets differed significantly from knockdown and knockout cell lines. Using GO ontology, we identified several critical biological processes associated specifically with catalytic activity and others attributable to non-catalytic interactions. Treatment of wild-type cells with KDAC-specific inhibitors Tubastatin A and PCI-34051 resulted in gene expression changes distinct from those of the engineered cell lines, validating this approach as a tool for evaluating in-cell inhibitor specificity and identifying off-target effects of KDAC inhibitors. Probing the functions of specific KDAC domains using these cell lines is not equivalent to doing so using previously existing methods and provides novel insight into the catalytic functions of individual KDACs by investigating the molecular and cellular changes upon genetic inactivation.

IntroductionNitrous oxide (N2O) emission from soil is a major concern due to its contribution to global climate change and its function as a loss mechanism of plant-available nitrogen (N) from the soil. This is especially true in intensive agricultural soils with high rates of N fertilizer application such as those on the semi-arid Southern High Plains, USA.MethodsThis study examined emissions of N2O, pore-space concentrations of N2O and nitric oxide (NO), soil chemical properties, water content, and the genetic potential for N cycling five years after conservation system and N management implementation.ResultsFor these semi-arid soils with low N, carbon, and water contents, large soil N2O emissions (up to 8 mL N2O-N m-2 day-1) are directly related to the application of N fertilizer which overwhelms the N2O reducing capacity of the soil. When this fertilizer N is depleted, N2O flux is either low, non-existent, or net-negative and has been observed as early as mid-season for preplant applied N fertilizer (-0.1 mL N2O-N m-2 day-1). Soil pore-space gas concentrations (N2O and NO) remained relatively constant across the growing season (average N2O: 0.78 µL N2O L-1 soil air; NO: 3.3 µL NO L-1 soil air, indicating a base-level of N-cycle activity, but was not directly related to surface emissions of N2O which decreased across the growing season. In addition, genetic potential for N cycle activities increased across the growing season simultaneously with stagnant/reduced N cycle activity. This reflects the difficulty in relating genetic potential to in-situ activity in field research.ConclusionIt is likely that in a nutrient and carbon-poor soil, such as the semi-arid agricultural soil in this study, the microbial processes associated with N cycling are mostly limited by inorganic-N and less directly related to genetic potential at the time of sampling.

Background: Head-loading, as a mode of transporting food, water and firewood, is a longstanding tradition assigned to female South African youth and has been associated with adverse health consequences. Objectives: This study determined the impact of head-loading on the neuromusculoskeletal health and proprioception of female South African youth. Method: This study comprised a counterbalanced, within-subject, single-factor experimental design which compared the changes that occurred when the same independent variable (head-loading) within two homogenous groups was measured in terms of the dependent variables (outcomes: neuromusculoskeletal pain and proprioception) at two time periods, before and after the introduction of the independent variable. A cohort of South African female youth (n = 100), aged 9–17 years, voluntarily partook in the study. The participants were randomly distributed into an experimental (n = 50) and a control (n = 50) group. The experimental group stood in a head-loaded state with their respective habitual head-load mass. Their proprioception measurements were compared during their unloaded versus loaded states, with the proprioceptive measurements including the total proprioception index, the anterior–posterior (front–back) index and the medial–lateral (side-to-side) index. Participants furthermore completed a head-loading health-related questionnaire. Results: Participants had a mean age of 12.3 ± 2.5 years, body mass of 44.4 ± 13.7 kg, stature of 145 ± 10 cm and a head-load mass of 8.0 ± 2.5 kg. Participants had poorer medial–lateral proprioception during head-loading as compared to their unloaded state (1.4 ± 0.8 as compared to 1.6 ± 0.9) (p 0.05). Most youth (96%) experienced neuromusculoskeletal pain in their cervical vertebrae (40.9%), shoulders (27.3%), lumbar vertebrae (10.7%), arms (8.3%), legs (8.3%), knees (1.9%), fingers (1.5%), toes (0.5%) and thoracic vertebrae (0.5%) (χ2: p 0.05). Conclusion: Head-loading adversely affects the medial–lateral proprioception and neuromusculoskeletal health of participants. Contribution: The findings of this study confirms that head-loading produces musculoskeletal pain.

Water bodies on Mars and the icy moons of the outer solar system are now recognized as likely being associated with high levels of salt. Therefore, the study of high salinity environments and their inhabitants has become increasingly relevant for Astrobiology. Members of the archaeal class Halobacteria are the most successful microbial group living in hypersaline conditions and are recognized as key model organisms for exposure experiments. Despite this, data for the class is uneven across taxa and widely dispersed across the literature, which has made it difficult to properly assess the potential for species of Halobacteria to survive under the polyextreme conditions found beyond Earth. Here we provide an overview of published data on astrobiology-linked exposure experiments performed with members of the Halobacteria, identifying clear knowledge gaps and research opportunities.

This study examined the effects of joint angle (JA) on maximal voluntary isometric contraction (MVIC) and neuromuscular responses following fatiguing tasks anchored to RPE. Nine men (mean ± SD: age = 20.7 ± 1.2 yrs) performed forearm flexion MVICs at elbow JAs of 75° and 125° before and after sustained, isometric forearm flexion tasks to failure at fatiguing joint angles (FJA) of 75° and 125° anchored to RPE = 8. The amplitude and frequency of the electromyographic and mechanomyographic signals were recorded. Neuromuscular efficiency was calculated by dividing normalized torque by normalized electromyographic amplitude. A dependent t-test was used to assess the mean difference for time to task failure (TTF) between FJA. Repeated measure ANOVAs were used to assess mean differences for pre-test to post-test MVIC and neuromuscular responses. There was no significant difference between FJA for TTF (p = 0.223). The MVIC (collapsed across FJA and MVIC JA) decreased from pre-test to post-test (51.1 ± 5.0 vs. 45.3 ± 5.6 Nm, p < 0.001). Normalized neuromuscular parameters remained unchanged (p > 0.05). The FJA resulted in similar torque and neuromuscular responses, and the decreases in MVIC were not tracked by changes in the neuromuscular parameters. Thus, the neuromuscular parameters were not sensitive to fatigue, and pre-test to post-test measures may be compared between different FJA.

The main objective of the current study was to perform a systematic literature review with the purpose of exploring the impact of long-chain n-3 polyunsaturated fatty acid (LCn-3 PUFA) relative to control oil supplementation on muscle strength, with secondary outcomes of muscle mass and physical function in older individuals under conditions of habitual physical activity/exercise. The review protocol was registered with PROSPERO (CRD42021267011) and followed the guidelines outlined in the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement. The search for relevant studies was performed utilizing databases such as PubMed, EMBASE, CINAHL, Scopus, Web of Science, and the Cochrane Central Register of Controlled Trials (CENTRAL) up to June 2023. Randomized controlled trials (RCTs) in older adults comparing the effects of LCn-3 PUFA with a control oil supplement on muscle strength were included. Five studies involving a total of 488 participants (348 females and 140 males) were identified that met the specified inclusion criteria and were included. Upon analyzing the collective data from these studies, it was observed that supplementation with LCn-3 PUFA did not have a significant impact on grip strength (standardized mean difference (SMD) 0.61, 95% confidence interval [−0.05, 1.27]; p = 0.07) in comparison to the control group. However, there was a considerable level of heterogeneity among the studies (I2 = 90%; p < 0.001). As secondary outcomes were only measured in a few studies, with significant heterogeneity in methods, meta-analyses of muscle mass and functional abilities were not performed. Papers with measures of knee extensor muscle mass as an outcome (n = 3) found increases with LCn-3 PUFA supplementation, but studies measuring whole body lean/muscle mass (n = 2) and functional abilities (n = 4) reported mixed results. With a limited number of studies, our data indicate that LCn-3 PUFA supplementation has no effect on muscle strength or functional abilities in older adults but may increase muscle mass, although, with only a few studies and considerable heterogeneity, further work is needed to confirm these findings.

It is evident that legume root nodules can accommodate rhizobial and non-rhizobial bacterial endophytes. Our recent nodule microbiome study in peanuts described that small nodules can harbor diverse bacterial endophytes. To understand their functional role, we isolated 87 indigenous endophytes from small nodules of field-grown peanut roots and characterized them at molecular, biochemical, and physiological levels. The amplified 16S rRNA genes and phylogenetic analysis of these isolates revealed a wide variety of microorganisms related to the genera Bacillus, Burkholderia, Enterobacter, Herbaspirillum, Mistsuaria, Pantoea, Pseudomonas, and Rhizobia. It was observed that 37% (100% identity) and 56% (>99% identity) of the isolates matched with the amplified sequence variants (ASVs) from our previous microbiome study. All of these isolates were tested for stress tolerance (high temperature, salinity, acidic pH) and phosphate (P) solubilization along with ammonia (NH3), indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate deaminase (ACCD), and siderophore production. The majority (78%) of the isolates were found to be halotolerant, thermotolerant, and acidophilic, and a few of them showed a significant positive response to the production of IAA, NH3, siderophore, ACCD, and P-solubilization. To evaluate the plant growth promotion (PGP) activity, plant and nodulation assays were performed in the growth chamber conditions for the selected isolates from both the non-rhizobial and rhizobial groups. However, these isolates appeared to be non-nodulating in the tested conditions. Nonetheless, the isolates 2 (Pantoea), 17 (Burkholderia), 21 (Herbaspirillum), 33o (Pseudomonas), and 77 (Rhizobium sp.) showed significant PGP activity in terms of biomass production. Our findings indicate that these isolates have potential for future biotechnological applications through the development of biologicals for sustainable crop improvement.

Abstract Background Sarcopenia is rarely diagnosed and is not recorded electronically in routine clinical care, posing challenges to trial recruitment. We describe the performance of four components of a strategy to efficiently recruit participants with sarcopenia to a trial of perindopril and/or leucine for sarcopenia: primary care vs. hospital recruitment, a comparison of central vs. local telephone pre‐screening, performance of a questionnaire on physical function conducted as part of the pre‐screening telephone call, and performance of bioimpedance measurement to identify low muscle mass. Methods Hospital‐based recruitment took place through inpatient and outpatient geriatric medicine services. Local research nurses reviewed medical notes and approached potentially eligible patients. Primary care recruitment reviewed primary care lists from collaborating practices, sending mailshots to patients aged 70 and over who were not taking angiotensin‐converting enzyme inhibitors. Telephone pre‐screening was conducted either by research nurses at each site or centrally by Tayside Clinical Trials Unit. The 10‐point SARC‐F questionnaire was used for pre‐screening. De‐identified recruitment information was held on a central electronic tracking system and analysed using SPSS. Bioimpedance was measured using the Akern BIA 101 system, with the Sergi equation used to estimate lean mass. Results Fourteen UK sites recruited to the trial. The 1202 sets of notes in hospital‐based care were reviewed at these sites; 7 participants (0.6% of total notes screened) were randomized. From primary care, 13 808 invitations were sent; 138 (1.0% of total invited) were randomized. 633/2987 primary care respondents were pre‐screened centrally; the mean number of calls per respondent was 2.3. For 10 sites where central and local pre‐screening could be compared, the conversion rate from pre‐screening to randomization was 18/588 (3.1%) for centralized calls, compared with 73/1814 (4.0%) for local pre‐screening calls (P = 0.29). A weak relationship was seen between higher (worse) SARC‐F score at screening and lower likelihood of progression to randomization (r = −0.08, P = 0.03). Muscle mass estimates generated using the Sergi equation were systematically biased, and a recalibrated equation for bioimpedance‐estimated muscle mass was derived. Conclusions Primary care recruitment led to higher response rates and overall numbers randomized than hospital‐based recruitment. Centralized pre‐screening saved local research nurses' time but did not improve conversion to randomization. SARC‐F did not help to target screening activity in this sarcopenia trial, and a recalibration of the equation for estimating muscle mass from bioimpedance measures may improve accuracy of the screening process.

Chimeric antigen receptor (CAR) T-cell immunotherapies targeting CD19 or CD22 induce remissions in the majority of patients with relapsed/refractory B-cell acute lymphoblastic leukemia (ALL), although relapse due to target antigen loss or downregulation has emerged as a major clinical dilemma. Accordingly, great interest exists in developing CAR T cells directed against alternative leukemia cell surface antigens that may help to overcome immunotherapeutic resistance. The fms-like tyrosine kinase 3 receptor (FLT3) is constitutively activated via FLT3 mutation in acute myeloid leukemia (AML) or wild-type FLT3 overexpression in KMT2A (lysine-specific methyltransferase 2A)-rearranged ALL, which are associated with poor clinical outcomes in children and adults. We developed monovalent FLT3-targeted CAR T cells (FLT3CART) and bispecific CD19xFLT3CART and assessed their anti-leukemia activity in preclinical models of FLT3-mutant AML and KMT2A-rearranged infant ALL. We report robust in vitro FLT3CART-induced cytokine production and cytotoxicity against AML and ALL cell lines with minimal cross-reactivity against normal hematopoietic and non-hematopoietic tissues. We also observed potent in vivo inhibition of leukemia proliferation in xenograft models of both FLT3-mutant AML and KMT2A-rearranged ALL, including a post-tisagenlecleucel ALL-to-AML lineage switch patient-derived xenograft model pairing. We further demonstrate significant in vitro and in vivo activity of bispecific CD19xFLT3CART against KMT2Arearranged ALL and posit that this additional approach might also diminish potential antigen escape in these high-risk leukemias. Our preclinical data credential FLT3CART as a highly effective immunotherapeutic strategy for both FLT3- mutant AML and KMT2A-rearranged ALL which is poised for further investigation and clinical translation.

Extreme weather events can temporarily alter the structure of coastal systems and generate floodwaters that are contaminated with fecal indicator bacteria (FIB); however, every coastal system is unique, so identification of trends and commonalities in these episodic events is challenging. To improve our understanding of the resilience of coastal systems to the disturbance of extreme weather events, we monitored water quality, FIB at three stations within Clear Lake, an estuary between Houston and Galveston, and three stations in bayous that feed into the estuary. Water samples were collected immediately before and after Hurricane Harvey (HH) and then throughout the fall of 2017. FIB levels were monitored by culturing E. coli and Enterococci. Microbial community structure was profiled by high throughput sequencing of PCR-amplified 16S rRNA gene fragments. Water quality and FIB data were also compared to historical data for these water body segments. Before HH, salinity within Clear Lake ranged from 9 to 11 practical salinity units (PSU). Immediately after the storm, salinity dropped to < 1 PSU and then gradually increased to historical levels over 2 months. Dissolved inorganic nutrient levels were also relatively low immediately after HH and returned, within a couple of months, to historical levels. FIB levels were elevated immediately after the storm; however, after 1 week, E. coli levels had decreased to what would be acceptable levels for freshwater. Enterococci levels collected several weeks after the storm were within the range of historical levels. Microbial community structure shifted from a system dominated by Cyanobacteria sp. before HH to a system dominated by Proteobacteria and Bacteroidetes immediately after. Several sequences observed only in floodwater showed similarity to sequences previously reported for samples collected following Hurricane Irene. These changes in beta diversity corresponded to salinity and nitrate/nitrite concentrations. Differential abundance analysis of metabolic pathways, predicted from 16S sequences, suggested that pathways associated with virulence and antibiotic resistance were elevated in floodwater. Overall, these results suggest that floodwater generated from these extreme events may have high levels of fecal contamination, antibiotic resistant bacteria and bacteria rarely observed in other systems.

Abstract Background In Australia, it is projected that one in four individuals will be at the nominal retirement age of 65 or over by 2056; this effect is expected to be especially pronounced in rural areas. Previous findings on the effects of retirement on wellbeing have been mixed. The present study explores the effects of employment and retirement on health and wellbeing among a sample of rural Australians. Methods Australian Rural Mental Health Study participants who were aged 45 or over (N = 2013) were included in a series of analyses to compare the health and wellbeing of individuals with differing employment and retirement circumstances. Self-reported outcome variables included perceived physical health and everyday functioning, financial wellbeing, mental health, relationships, and satisfaction with life. Results Across the outcomes, participants who were employed or retired generally reported better health and wellbeing than those not in the workforce. Retired participants rated more highly than employed participants on mental health, relationships, and satisfaction with life. There was also a short-term benefit for perceived financial status for retired participants compared to employed participants, but this effect diminished over time. Conclusions While retirement is a significant life transition that may affect multiple facets of an individual’s life, the direction and magnitude of these effects vary depending on the retirement context, namely the pre-retirement and concurrent circumstances within which an individual is retiring. Personal perceptions of status changes may also contribute to an individual’s wellbeing more so than objective factors such as income. Policies that promote rural work/retirement opportunities and diversity and address rural disadvantage are needed.

Abstract Recognition and classification of mosquitoes is a critical component of vector-borne disease management. Vector surveillance, based on wingbeat frequency and other parameters, is becoming increasingly important in the development of automated identification systems, but inconsistent data quality and results frequently emerge from different techniques and data processing methods which have not been standardized on wingbeat collection of numerous species. We developed a simple method to detect and record mosquito wingbeat by multi-dimensional optical sensors and collected 21,825 wingbeat files from 29 North American mosquito species. In pairwise comparisons, wingbeat frequency of twenty six species overlapped with at least one other species. No significant differences were observed in wingbeat frequencies between and within individuals of Culex quinquefasciatus over time. This work demonstrates the potential utility of quantifying mosquito wingbeat frequency by infrared light sensors as a component of an automated mosquito identification system. Due to species overlap, wingbeat frequency will need to integrate with other parameters to accurately delineate species in support of efficient mosquito surveillance, an important component of disease intervention.

The present study examined the effect of anchor schemes on the time to task failure (TTF), performance fatigability, neuromuscular responses, and the perceived sensations that contributed to task termination following the sustained, isometric forearm flexion tasks. Eight women completed sustained, isometric forearm flexion tasks anchored to RPE = 8 (RPEFT) and the torque (TRQFT) that corresponded to RPE = 8. The subjects performed pre-test and post-test maximal isometric contractions to quantify performance fatigability and changes in electromyographic amplitude (EMG AMP) and neuromuscular efficiency (NME). In addition, the subjects completed a post-test questionnaire (PTQ) to quantify the contributions of perceived sensations to task termination. Repeated measure ANOVAs were used to assess the mean differences for TTF, performance fatigability, and neuromuscular responses. Wilcoxon Signed Rank Tests were used to assess the differences between anchor schemes for the average values from the PTQ item scores. For TTF, the RPEFT was longer than the TRQFT (174.9 ± 85.6 vs. 65.6 ± 68.0 s; p = 0.006). Collapsed across the anchor scheme, there were decreases in torque (23.7 ± 5.5 Nm vs. 19.6 ± 4.9 Nm; p < 0.001) and NME (1.00 ± 0.00 vs. 0.76 ± 0.15; p = 0.003). There were no significant (p > 0.577) changes for EMG AMP. For the PTQ, there were no differences (p > 0.05) between anchor schemes. There were, however, inter-individual differences in the response scores. The current findings indicated that performance fatigability was likely due to peripheral fatigue (based on NME), not central fatigue (based on EMG AMP). Furthermore, the use of a PTQ may serve as a simple tool to assess the contributions of perceived sensations to task termination.