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Diazotrophic bacteria can reduce atmospheric nitrogen into ammonia enabling bioavailability of the essential element. Many diazotrophs closely associate with plant roots increasing nitrogen availability, acting as plant growth promoters. These associations have the potential to reduce the need for costly synthetic fertilizers if they could be engineered for agricultural applications. However, despite the importance of diazotrophic bacteria, genetic tools are poorly developed in a limited number of species, in turn narrowing the crops and root microbiomes that can be targeted. Here, we report optimized protocols and plasmids to manipulate phylogenetically diverse diazotrophs with the goal of enabling synthetic biology and genetic engineering. Three broad-host-range plasmids can be used across multiple diazotrophs, with the identification of one specific plasmid (containing origin of replication RK2 and a kanamycin resistance marker) showing the highest degree of compatibility across bacteria tested. We then demonstrated modular expression by testing seven promoters and eleven ribosomal binding sites using proxy fluorescent proteins. Finally, we tested four small molecule inducible systems to report expression in three diazotrophs and demonstrated genome editing in Klebsiella michiganensis M5al.One-sentence summaryIn this study, broad-host plasmids and synthetic genetic parts were leveraged to enable expression tools in a library of diazotrophic bacteria.

Fugitive methane (CH4) emissions occur in the whole chain of oil and gas production, including from extraction, transportation, storage, and distribution. Such emissions are usually detected and quantified by conducting surveys as close as possible to the source location. However, these surveys are labour-intensive, are costly, and fail to not provide continuous emissions monitoring. The deployment of permanent sensor networks in the vicinity of industrial CH4 emitting facilities would overcome the limitations of surveys by providing accurate emission estimates, thanks to continuous sampling of emission plumes. Yet high-precision instruments are too costly to deploy in such networks. Low-cost sensors using a metal oxide semiconductor (MOS) are presented as a cheap alternative for such deployments due to their compact dimensions and to their sensitivity to CH4. In this study, we demonstrate the ability of two types of MOS sensors (TGS 2611-C00 and TGS 2611-E00) manufactured by Figaro® to reconstruct a CH4 signal, as measured by a high-precision reference gas analyser, during a 7 d controlled release campaign conducted by TotalEnergies® in autumn 2019 near Pau, France. We propose a baseline voltage correction linked to atmospheric CH4 background variations per instrument based on an iterative comparison of neighbouring observations, i.e. data points. Two CH4 mole fraction reconstruction models were compared: multilayer perceptron (MLP) and second-degree polynomial. Emission estimates were then computed using an inversion approach based on the adjoint of a Gaussian dispersion model. Despite obtaining emission estimates comparable with those obtained using high-precision instruments (average emission rate error of 25 % and average location error of 9.5 m), the application of these emission estimates is limited to adequate environmental conditions. Emission estimates are also influenced by model errors in the inversion process.

Abstract Common beans are a common staple food with valuable nutritional qualities, but their high contents in antinutritional factors (ANFs) can decrease the bioavailability of (i) fat-soluble micronutrients including carotenoids and (ii) minerals. Our objective was to select ANF-poor bean lines that would not interfere with carotenoid and mineral bioavailability. To achieve this objective, seeds of commercial and experimental Phaseolus vulgaris L. bean lines were produced for 2 years and the bean’s content in ANFs (saponins, phytates, tannins, total polyphenols) was assessed. We then measured carotenoid bioaccessibility and mineral solubility (i.e. the fraction of carotenoid and mineral that transfer into the aqueous phase of the digesta and is therefore absorbable) from prepared beans using in vitro digestion. All beans contained at least 200 mg/100 g of saponins and 2.44 mg/100 g tannins. The low phytic acid (lpa) lines, lpa1 and lpa12 exhibited lower phytate levels (≈ − 80%, p = 0.007 and p = 0.02) than their control BAT-93. However, this decrease had no significant impact on mineral solubility. HP5/1 (lpa + phaseolin and lectin PHA-E free) bean line, induced an improvement in carotenoid bioaccessibility (i.e., + 38%, p = 0.02, and + 32%, p = 0.005, for phytofluene bioaccessibility in 2021 and 2022, respectively). We conclude that decrease in the phytate bean content should thus likely be associated to decreases in other ANFs such as tannins or polyphenols to lead to significant improvement of micronutrient bioaccessibility.

Hosting capacity (HC) describes the electricity network’s ability to accommodate distributed generation (DG) without deteriorating electrical performance indicators. Distribution system operators typically express their networks’ HC as a single threshold, called static hosting capacity (SHC). SHC is determined via conservative regulatory criteria, increasing connection costs and time. This paper explores the potential for additional energy injection into the network via dynamic hosting capacity (DHC). A network node’s DHC is derived from the hourly operation of the network, accounting for the time variability of existing distributed generation (DG) output and demand. The methodology considers the network assets’ N-1 contingencies and their probabilities, defining the security-aware DHC (SDHC). The SDHC definition is technologically neutral. Through a case study of a radial medium voltage distribution network, the paper highlights the significant limitations of SHC due to conservative calculation criteria mandated by regulators. Annual injectable energy is increased by 62% to 76% when comparing DHC to SHC. Variations between average DHC and SDHC are below 0.01% due to low N-1 probabilities. This finding points out the potential of dynamic hosting capacity definitions, allowing more efficient use of the existing network and facilitating the integration of new DG capacity with reduced connection costs and time.

Peptide CRF antagonists injected peripherally alleviate stress-induced visceral hypersensitivity (SIVH) to colorectal distension (CRD) in rodents. Here we further evaluated the dose and time-dependent inhibitory activity of several long-acting peptide CRF receptor antagonists related to astressin on SIVH, focusing on astressin C (AstC), which previously showed high efficacy on stress-related alterations of HPA axis and gut secretomotor functions. Male and female Sprague-Dawley rats pretreated subcutaneously (SC) with AstC were injected intraperitoneally (IP) with CRF 15 min later. The visceromotor responses (VMR) to graded phasic CRD (10, 20, 40 and 60 mmHg) were monitored at basal, 15 min and up to 1-8 days after pretreatment. Two other astressin analogs, hexanoyl-astressin D (Hex-AstD) and [CαMeVal19,32]-AstC, were also tested. The response to IP CRF was sex-dependent with female rats requiring a higher dose to exhibit visceral hyperalgesia. Pretreatment with AstC (30-1000 µg/kg) resulted in a dose-related inhibition of IP CRF-induced SIVH and diarrhea in both sexes. The highest dose prevented SIVH and diarrhea up to 5-7 days after a single SC injection and was lost on day 7 (females) and day 8 (males) but reinstated after a second injection of AstC on day 8 or 9 respectively. [CαMeVal19,32]-AstC and Hex-AstD (1000 µg/kg in males) also prevented SIVH. These data show the potent long-lasting anti-hyperalgesic effect of AstC in an acute model of SIVH in both male and female rats. This highlights the potential of long-acting peripheral CRF antagonists to treat stress-sensitive irritable bowel syndrome.

Abstract Background Duchenne muscular dystrophy (DMD) is an X‐linked disorder characterized by progressive muscle weakness due to the absence of functional dystrophin. DMD patients also develop dilated cardiomyopathy (DCM). We have previously shown that DMD (mdx) mice and a canine DMD model (GRMD) exhibit abnormal intracellular calcium (Ca2+) cycling related to early‐stage pathological remodelling of the ryanodine receptor intracellular calcium release channel (RyR2) on the sarcoplasmic reticulum (SR) contributing to age‐dependent DCM. Methods Here, we used hiPSC‐CMs from DMD patients selected by Speckle‐tracking echocardiography and canine DMD cardiac biopsies to assess key early‐stage Duchenne DCM features. Results Dystrophin deficiency was associated with RyR2 remodelling and SR Ca2+ leak (RyR2 Po of 0.03 ± 0.01 for HC vs. 0.16 ± 0.01 for DMD, P

IntroductionThe 21-point Brain Care Score (BCS) is a novel tool designed to motivate individuals and care providers to take action to reduce the risk of stroke and dementia by encouraging lifestyle changes. Given that late-life depression is increasingly recognized to share risk factors with stroke and dementia, and is an important clinical endpoint for brain health, we tested the hypothesis that a higher BCS is associated with a reduced incidence of future depression. Additionally, we examined its association with a brain health composite outcome comprising stroke, dementia, and late-life depression.MethodsThe BCS was derived from the United Kingdom Biobank baseline evaluation in participants with complete data on BCS items. Associations of BCS with the risk of subsequent incident late-life depression and the composite brain health outcome were estimated using multivariable Cox proportional hazard models. These models were adjusted for age at baseline and sex assigned at birth.ResultsA total of 363,323 participants were included in this analysis, with a median BCS at baseline of 12 (IQR: 11-14). There were 6,628 incident cases of late-life depression during a median follow-up period of 13 years. Each five-point increase in baseline BCS was associated with a 33% lower risk of incident late-life depression (95% CI: 29%-36%) and a 27% lower risk of the incident composite outcome (95% CI: 24%-30%).DiscussionThese data further demonstrate the shared risk factors across depression, dementia, and stroke. The findings suggest that a higher BCS, indicative of healthier lifestyle choices, is significantly associated with a lower incidence of late-life depression and a composite brain health outcome. Additional validation of the BCS is warranted to assess the weighting of its components, its motivational aspects, and its acceptability and adaptability in routine clinical care worldwide.

Background The implementation of preventive therapies among patients with stroke remains inadequately explored, especially when compared with patients with myocardial infarction (MI), despite sharing similar vascular risk profiles. We tested the hypothesis that participants with a history of stroke have a worse cardiovascular prevention profile in comparison to participants with MI. Methods and Results In cross‐sectional analyses within the UK Biobank and All of Us Research Program, involving 14 760 (9193 strokes, 5567 MIs) and 7315 (2948 strokes, 4367 MIs) participants, respectively, we evaluated cardiovascular prevention profiles assessing low‐density lipoprotein (

Ex-utero regulation of the lungs' responses to breathing air and continued alveolar development shape adult respiratory health. Applying single-cell transposome hypersensitive site sequencing (scTHS-seq) to over 80,000 cells, we assembled the first regulatory atlas of postnatal human and mouse lung alveolar development. We defined regulatory modules and elucidated new mechanistic insights directing alveolar septation, including alveolar type 1 and myofibroblast cell signaling and differentiation, and a unique human matrix fibroblast population. Incorporating GWAS, we mapped lung function causal variants to myofibroblasts and identified a pathogenic regulatory unit linked to lineage marker FGF18, demonstrating the utility of chromatin accessibility data to uncover disease mechanism targets. Our regulatory map and analysis model provide valuable new resources to investigate age-dependent and species-specific control of critical developmental processes. Furthermore, these resources complement existing atlas efforts to advance our understanding of lung health and disease across the human lifespan.

In his comment (Kowalski 2023) on our recent publication (Heiskanen et al. 2022) where we present the Integrated Carbon Observation System (ICOS) research infrastructure, Andrew Kowalski introduces three important and, [...]

Background Vascular brain injury (VBI) may be an under-recognised contributor to mobility impairment. We examined associations between MRI VBI biomarkers and impaired mobility.Methods We separately analysed Atherosclerosis Risk in Communities (ARIC) and UK Biobank (UKB) study cohorts. Inclusion criteria were no prevalent clinical stroke, and available brain MRI and balance and gait data. MRI VBI biomarkers were (ARIC: ventricular and white matter hyperintensity (WMH) volumes, non-lacunar and lacunar infarctions, microhaemorrhage; UKB: ventricular, brain and WMH volumes, fractional anisotropy (FA), mean diffusivity (MD), intracellular and isotropic free water volume fractions). Quantitative biomarkers were categorised into tertiles. Mobility impairment outcomes were imbalance and slow walk in ARIC and recent fall and slow walk in UKB. Adjusted multivariable logistic regression analyses were performed.Results We included 1626 ARIC (mean age 76.2 years; 23.4% imbalance, 25.0% slow walk) and 40 098 UKB (mean age 55 years; 15.8% falls, 2.8% slow walk) participants. In ARIC, imbalance associated with four of five VBI measures (all p values

Background Patients with ischemic stroke and concomitant COVID‐19 infection have worse outcomes than those without this infection, but the impact of COVID‐19 on hemorrhagic stroke remains unclear. We aimed to assess if COVID‐19 worsens outcomes in intracerebral hemorrhage (ICH). Methods and Results We conducted an observational study of ICH outcomes using Get With The Guidelines Stroke data. We compared patients with ICH who were COVID‐19 positive and negative during the pandemic (March 2020–February 2021) and prepandemic (March 2019–February 2020). Main outcomes were poor functional outcome (defined as a modified Rankin scale score of 4 to 6 at discharge), mortality, and discharge to a skilled nursing facility or hospice. The first stage included 60 091 patients with ICH who were COVID‐19 negative and 1326 COVID‐19 positive. In multivariable analyses, patients with ICH with versus without COVID‐19 infection had 68% higher odds of poor outcome (odds ratio [OR], 1.68 [95% CI, 1.41–2.01]), 51% higher odds of mortality (OR, 1.51 [95% CI, 1.33–1.71]), and 66% higher odds of being discharged to a skilled nursing facility/hospice (OR, 1.66 [95% CI, 1.43–1.93]). The second stage included 62 743 prepandemic and 64 681 intrapandemic cases with ICH. In multivariable analyses, patients with ICH admitted during versus before the COVID‐19 pandemic had 10% higher odds of poor outcomes (OR, 1.10 [95% CI, 1.07–1.14]), 5% higher mortality (OR, 1.05 [95% CI, 1.02–1.08]), and no significant difference in the risk of being discharged to a skilled nursing facility/hospice (OR, 0.93 [95% CI, 0.90–0.95]). Conclusions The pathophysiology of the COVID‐19 infection and changes in health care delivery during the pandemic played a role in worsening outcomes in the patient population with ICH.

Increased blood pressure variability (BPV) is linked to cardiovascular disease and mortality, yet few modifiable BPV risk factors are known. We aimed to assess the relationship between sleep quality and activity level on longitudinal BPV in a cohort of community-dwelling adults (age ≥18) from 17 countries. Using Withings home measurement devices, we examined sleep quality and physical activity over one year, operationalized as mean daily step count and number of sleep interruptions, both transformed into tertiles. The primary study outcome was high BPV, defined as the top tertile of systolic blood pressure standard deviation. Our cohort comprised 29,375 individuals (mean age = 58.6 years) with 127.8±90.1 mean days of measurements. After adjusting for age, gender, country, body mass index, measurement days, mean blood pressure, and total time in bed, the odds ratio of having high BPV for those in the top tertile of sleep interruptions (poor sleep) was 1.37 (95% CI, 1.28-1.47) and 1.44 (95% CI, 1.35-1.54) for those in the lowest tertile of step count (physically inactive). Combining these exposures revealed a significant excess relative risk of 0.20 (95% CI, 0.04-0.35, p = 0.012), confirming their super-additive effect. Comparing individuals with the worst exposure status (lowest step count and highest sleep interruptions, n = 2,690) to those with the most optimal status (highest step count and lowest sleep interruptions, n = 3,531) yielded an odds ratio of 2.01 (95% CI, 1.80-2.25) for high BPV. Our findings demonstrate that poor sleep quality and physical inactivity are associated with increased BPV both independently and super-additively.

Background The American Heart Association's Life's Simple 7, a public health construct capturing key determinants of cardiovascular health, became the Life's Essential 8 after the addition of sleep duration. The authors tested the hypothesis that suboptimal sleep duration is associated with poorer neuroimaging brain health profiles in asymptomatic middle‐aged adults. Methods and Results The authors conducted a prospective magnetic resonance neuroimaging study in middle‐aged individuals without stroke or dementia enrolled in the UK Biobank. Self‐reported sleep duration was categorized as short (

IntroductionThe 21-point Brain Care Score (BCS) was developed through a modified Delphi process in partnership with practitioners and patients to promote behavior changes and lifestyle choices in order to sustainably reduce the risk of dementia and stroke. We aimed to assess the associations of the BCS with risk of incident dementia and stroke.MethodsThe BCS was derived from the United Kingdom Biobank (UKB) baseline evaluation for participants aged 40–69 years, recruited between 2006–2010. Associations of BCS and risk of subsequent incident dementia and stroke were estimated using Cox proportional hazard regressions, adjusted for sex assigned at birth and stratified by age groups at baseline.ResultsThe BCS (median: 12; IQR:11–14) was derived for 398,990 UKB participants (mean age: 57; females: 54%). There were 5,354 incident cases of dementia and 7,259 incident cases of stroke recorded during a median follow-up of 12.5 years. A five-point higher BCS at baseline was associated with a 59% (95%CI: 40-72%) lower risk of dementia among participants aged 59 years. A five-point higher BCS was associated with a 48% (95%CI: 39-56%) lower risk of stroke among participants aged 59.DiscussionThe BCS has clinically relevant and statistically significant associations with risk of dementia and stroke in approximately 0.4 million UK people. Future research includes investigating the feasibility, adaptability and implementation of the BCS for patients and providers worldwide.

Detecting and quantifying CH4 gas emissions at industrial facilities is an important goal for being able to reduce these emissions. The nature of CH4 emissions through “leaks” is episodic and spatially variable, making their monitoring a complex task; this is partly being addressed by atmospheric surveys with various types of instruments. Continuous records are preferable to snapshot surveys for monitoring a site, and one solution would be to deploy a permanent network of sensors. Deploying such a network with research-level instruments is expensive, so low-cost and low-power sensors could be a good alternative. However, low cost usually entails lower accuracy and the existence of sensor drifts and cross-sensitivity to other gases and environmental parameters. Here we present four tests conducted with two types of Figaro® Taguchi gas sensors (TGSs) in a laboratory experiment. The sensors were exposed to ambient air and peaks of CH4 concentrations. We assembled four chambers, each containing one TGS sensor of each type. The first test consisted in comparing parametric and non-parametric models to reconstruct the CH4 peak signal from observations of the voltage variations of TGS sensors. The obtained relative accuracy is better than 10 % to reconstruct the maximum amplitude of peaks (RMSE ≤2 ppm). Polynomial regression and multilayer perceptron (MLP) models gave the highest performances for one type of sensor (TGS 2611C, RMSE =0.9 ppm) and for the combination of two sensors (TGS 2611C + TGS 2611E, RMSE =0.8 ppm), with a training set size of 70 % of the total observations. In the second test, we compared the performance of the same models with a reduced training set. To reduce the size of the training set, we employed a stratification of the data into clusters of peaks that allowed us to keep the same model performances with only 25 % of the data to train the models. The third test consisted of detecting the effects of age in the sensors after 6 months of continuous measurements. We observed performance degradation through our models of between 0.6 and 0.8 ppm. In the final test, we assessed the capability of a model to be transferred between chambers in the same type of sensor and found that it is only possible to transfer models if the target range of variation of CH4 is similar to the one on which the model was trained.

At operational level, fossil fuel phase-out and high shares of non-dispatchable renewable energy resources (RES) will challenge the system operator’s (SO) ability to balance generation, and the demand at any time. The variability of RES output ranges from one hour to a season, and critical events such as low supply and high demand might occur more frequently and for more extended periods. When evaluating the role of Energy Storage Systems (ESSs) in this context, the need for a long time scope to capture the different RES variabilities must be reconciled with the need for modeling the hourly chronology. This paper presents a medium-term operation planning model, addressing both the energy dispatch and the balancing services. This study shows that representing the combined chronological variability of demand and RES production is essential to properly assess the roles of different kinds of ESSs in the future 2030 electricity mix. Otherwise, it would not be possible to appropriately capture the frequency, depth, and length of events for which ESSs are activated. The analysis also highlights the importance of considering balancing services, given the significant contribution of batteries to the reserve market. Finally, the results show that batteries and Pumped Storage Hydro (PSH) have different roles in the Spanish electricity system with a high renewable penetration. While PSH is mainly used to provide energy during critical periods, batteries mostly provide balancing services.

Provision of firm capacity will become a challenge in power systems dominated by renewable generation. This paper analyzes the competitiveness and role of battery storage, six types of pumped-hydro storage, open cycle gas turbine (OCGT), and demand response (DR) technologies in providing the firm capacity required to guarantee the security of supply in a real-size power system such as the Spanish one in horizon 2030. The paper contributes with detailed and realistic modeling of the DR capabilities. Demand is disaggregated by sector and activities and projected towards 2030, applying a growth rate by activity. The load flexibility constraints are considered to ensure the validity of the results. A generation operation planning and expansion model, SPLODER, is conveniently upgraded to properly represent the different storage alternatives addressed in the paper. The results highlight the importance of considering demand response for evaluating long-term firm capacity requirements, showing a non-negligible impact on the investment decisions on the amount of firm capacity required in the system and the optimal shares of wind and solar PV renewable generation. Results also show the dominance of cost-competitiveness of pumped hydro and OCGTs over batteries. Additionally, capacity payments are required to support firm capacity providers’ investments.