However, if the SLR rate accelerates beyond the accretion ability of the wetland, a significant shift in species composition and an increase in open water coverage was predicted. These results indicate that the current rate of sediment capture by wetland species, and the subsequent rate of elevation change, will need to increase significantly to adapt with projected future rates of SLR.The dictyochophyte microalga Pseudochattonella verruculosa was responsible for the largest farmed fish mortality ever recorded in the world, with losses for the Chilean salmon industry amounting to US$ 800 M in austral summer 2016. Super-scale climatic anomalies resulted in strong vertical water column stratification that stimulated development of a dynamic P. verruculosa thin layer (up to 38 μg chl a L-1) for several weeks in Reloncaví Sound. Hydrodynamic modeling (MIKE 3D) indicated that the Sound had extremely low flushing rates (between 121 and 200 days) in summer 2016. Reported algal cell densities of 7000-20,000 cells mL-1 generated respiratory distress in fish that was unlikely due to low dissolved oxygen (permanently >4 mg L-1). Histological examination of salmon showed that gills were the most affected organ with significant tissue damage and circulatory disorders. It is possible that some of this damage was due to a diatom bloom that preceded the Pseudochattonella event, thereby rendering the fish m bloom. Multiple mitigation strategies were used by salmon farmers during the event, with only delayed seeding of juvenile fish into the cages and towing of cages to sanctuary sites being effective. Airlift pumping, used effectively against other fish-killing HABs in the US and Canada was not effective, perhaps because it brought subsurface layers of Pseudochattonella to the surface, or and it also may have lysed the fragile cells, rendering them more lethal. The present study highlights knowledge gaps and inefficiency of contingency plans by the fish farming industry to overcome future fish-killing algal blooms under future climate change scenarios. The use of new technologies based on molecular methods for species detection, good farm practices by fish farms, and possible mitigation strategies are discussed.The transboundary Tigris-Euphrates Basin (TEB) is prone to water-scarcity disputes. Water scarcity is related to aridity, climate extremes, limited supplies, upstream reservoir storage, rising water demand, and population growth. Understanding the water budget and storage changes in the basin in relation to hydrological extremes is fundamental to mitigate the drought and flood impacts and the key to efficient water resources management. This study evaluated the water budget related to drought occurrences in the TEB over four decades (1979-2020) based on GRACE/GRACE-FO, and altimetry satellites data, in situ observations, and hydrological modeling using a Bayesian model averaging (BMA) approach. Results show that severe droughts occurred at about decadal timescales with increasing recovery times. Severe and exceptional droughts dominated from (1998 to 2000, 2007 to 2009). Mild to moderate droughts occurred in 1983-1984, 1989-1992, 2011-2013, and 2018. The most severe drought occurred in 2007-2009, with the largest decline (-80 km3) in GRACE total water storage (TWS). Depletion in TWS was dominated by depletion in reservoir storage. In contrast, groundwater (GW) depletion accounted for only 25-30% of TWS decline. Storage depletion was amplified by human intervention (e.g., irrigation and GW abstraction) by at least 50% during drought. Marked recovery in TWS occurred in 2019 and 2020 (totaling ~144 km3 by July 2020, representing ~2× total depletion between 2007 and 2018) in response to regional flooding. Applying the BMA approach to the estimates of water cycle fluxes improved the accuracy and similarity of storage change, but not variability relative to GRACE. In summary, prolonged droughts are the norm rather than the exception in the TEB over the past four decades. The frequency and severity of droughts have substantial implications for water scarcity for countries sharing the TEB and underscore riparian countries’ needs to expand their water management portfolio to mitigate drought impacts.The broad application of strobilurin fungicide led to pathogen resistance, and toxic effects have been reported for several species. Benzene kresoxim-methyl (BKM) is a novel strobilurin fungicide mainly used to control the cucumber powdery mildew. However, information about the fate of BKM in agrofood systems and related human exposure is limited. In this study, greenhouse experiments were conducted to investigate the distribution, translocation, and residual of the 10% suspension concentrate (SC) commercial BKM formulations on mature cucumber plants using 14C tracer technology. After foliage and fruit application, 25.84% of the applied 14C-labeled BKM can be absorbed into mature cucumber plants at 21 days after treatment. The absorbed BKM transferred throughout the plant acropetally and basipetally, although over 81.13% of absorbed BKM remained in the labeled leaves. In the edible parts, 14.35% of the absorbed BKM remained in the pericarp of labeled fruits, only 0.027 mg kg-1 accumulated in the sarcocarp. The concentration of BKM in newborn fruits was 0.005 mg kg-1, indicating low dietary exposure. These findings develop a better understanding of the fate of BKM in the cucumber plants, provide guidance in the rational use of BKM and can be incorporated into food and environmental assessments of BKM.Non-exhaust emissions (NEE) of particulate matter (PM) from brake, tyre, road pavement and railway wear, as well as resuspension of already deposited road dust, account for up to 90% by mass of total traffic-related PM emitted. This review aims at analysing the current knowledge on road traffic NEE regarding sources, particle generation processes, chemical and physical characterization, and mitigation strategies. The literature on this matter often presents highly variable and hardly comparable results due to the heterogeneity of NEE sources and the absence of standardized sampling and measurement protocols. As evidence, emission factors (EFs) were found to range from 1 mg km-1 veh-1 to 18.5 mg km-1 veh-1 for brake wear, and from 0.3 mg km-1 veh-1 to 7.4 mg km-1 veh-1 for tyre wear. Resuspended dust, which varies in even wider ranges (from 5.4 mg km-1 veh-1 to 330 mg km-1 veh-1 for cars), is considered the prevailing NEE source. The lack of standardized monitoring approaches resulted in the impossibility of setting international regulations to limit NEE. Therefore, up until now the abatement of NEE has only been achieved by mitigation and prevention strategies. However, the effectiveness of these measures still needs to be improved and further investigated. As an example, mitigation strategies, such as street washing or sweeping, proved effective in reducing PM levels, but only in the short term. The replacement of internal combustion engines vehicles with electric ones was instead proposed as a prevention strategy, but there are still concerns regarding the increase of NEE deriving from the extra weight of the batteries. The data reported in this review highlighted the need for future studies to broaden their research area, and to focus not only on the standardization of methods and the introduction of regulations, but also on improving already existing technologies and mitigating strategies.Seasonal hydrological variation and chemical pollution represent two main drivers of freshwater biodiversity change in Mediterranean rivers. We investigated to what extent low flow conditions can modify the effects of chemical pollution on macroinvertebrate communities. To that purpose, we selected twelve sampling sites in the upper Tagus river basin (central Spain) having different sources of chemical pollution and levels of seasonal hydrological variation. The sites were classified as natural (high flow variation, low chemical impact), agricultural (high flow variation, high agricultural chemical inputs) and urban (limited flow variation, high urban chemical inputs). In these sites, we measured daily water discharge, nutrients, and contaminant concentrations, and we sampled benthic macroinvertebrates, in spring, summer and autumn. Significant differences related to toxic pressure and nutrient concentrations were observed between the three groups of sites. Seasonal patterns were found for some water quality parameters (e.g. nitrites, ammonia, suspended solids, metal toxicity), particularly in agricultural sites. Taxonomic and functional richness were slightly lower in the polluted sites (agricultural and urban), particularly during low flow periods (summer and early autumn). Functional diversity was significantly lower in sites with seasonal flow variation (agricultural sites) as compared to the more constant ones (urban sites). The frequency of traits such as large size, asexual reproduction, aquatic passive dispersion and the production of cocoons increased in response to pollution during low flow periods. This study shows that the impacts of anthropogenic chemical pollution on taxonomic and functional characteristics of macroinvertebrate communities seem to be larger during low flow periods. Therefore, further studies and monitoring campaigns assessing the effects of chemical pollution within these periods are recommended.This study investigated the sex-related and spatial variation in trace elements concentrations in feathers of the Kentish plover (Charadrius alexandrinus) inhabiting different coastal locations in the Iberian Peninsula. Feathers were sampled in a total of 50 breeding birds from 7 different locations 3 on the Mediterranean coast (n=18), 1 on the Atlantic coast of Andalusia (n=9) and 3 on the NW Atlantic coast (n=23). The feathers were analyzed to determine the concentrations of Al, Cr, Mn, Cu, Zn, As, Se, Cd, Hg and Pb. All of the trace elements were present at detectable concentrations, and the mean concentrations of Al, Zn and Cu were highest of those considered. The coastal section had a significant effect on 5 elements (Mn, Cu, As, Cd and Pb). The mean concentrations of almost all of the trace elements were highest in the birds from Andalusia, intermediate in the birds from the Mediterranean area and, in general, lowest in the birds from the NW Atlantic coast. Females showed significant higher Mn concentrations and high levels of exogenous elements (Al and As) in contrast with males. Less preening activity during the day of female birds, when they are occupied in incubating eggs, could favour higher concentrations of exogenous trace elements. The high levels of several trace elements observed in the samples from Mediterranean and Atlantic coast of Andalusia may pose a serious threat to these populations.Nanoplastics are a growing environmental and public health concern. However, the toxic mechanisms of nanoplastics are poorly understood. Here, we evaluated the effects of spherical polystyrene nanoplastics on reproduction of Daphnia pulex and analyzed the proteome of whole animals followed by molecular and biochemical analyses for the development of an adverse outcome pathway (AOP) for these contaminants of emerging concern. Animals were exposed to polystyrene nanoplastics (0, 0.1, 0.5, 1 and 2 mg/L) via water for 21 days. Nanoplastics negatively impacted cumulative offspring production. A total of 327 differentially expressed proteins (DEPs) were identified in response to nanoplastics which were further validated from gene expression and enzyme activity data. Based on these results, we propose an AOP for nanoplastics, including radical oxygen species production and oxidative stress as the molecular initiating event (MIE); followed by changes in specific signaling pathways (Jak-STAT, mTOR and FoxO) and in the metabolism of glutathione, protein, lipids, and molting proteins; with an end result of growth inhibition and decrease reproductive output. This study serves as a foundation for the development of a mechanistic understanding of nanoplastic toxicity in crustaceans and perhaps other aquatic organisms.Soil respiration (RS) from cropland in response to tillage practices contribute to global climate change. We quantified the effect of no-tillage (NT) and conventional tillage (CT) on RS and precipitation in the North China Plain (NCP). An in-situ automatic sampling and measurement method was applied during the maize (Zea mays L.) growth stages in 2018 and 2019. The continuous daily RS, soil water content and temperature were monitored during all the maize growth stages, whereas maize grain yield, aboveground biomass, and soil microbial biomass were measured after harvest. The mean RS across tillage practices on bright days was higher in 2018 (16.69 g CO2 m-2 d-1) than that in 2019 (12.99 g CO2 m-2 d-1). Compared with CT, NT increased RS on bright days by 31.44% in 2018 and 15.60% in 2019. However, mean RS on rain-affected days across tillage practices was lower in 2018 than that in 2019. NT increased mean RS after precipitation in 2018 (p 0.05). Overall, influence of tillage practices and precipitation on RS were different according to soil water content. Therefore, it is necessary to decrease excessive irrigation to reduce RS in dry years and to conduct continuous observations on RS after precipitation in the NCP.White-winged scoters (Melanitta fusca; WWSC) and surf scoters (M. perspicillata; SUSC) have declined by over 60% in recent decades. Identifying contributing factors from within a mosaic of sublethal, multiple stressors is challenging. In urbanized Puget Sound, Washington, USA where scoters winter, changes in prey availability explained only a portion of local declines, suggesting that other „silent stressors” such as sublethal contaminants might play a role. Past studies of pollutant effects on scoters used Fisherian statistics that often revealed few correlates; however, novel statistical approaches could detect and provide more insights about sublethal impacts. Our objectives were to (1) relate pollutant accumulation to health of the birds, and (2) compare permutational multivariate statistics with traditional approaches in identifying sublethal health effects. We collected scoters from three locations in Puget Sound in December 2005 and March 2006, and measured cadmium (Cd), mercury (Hg), and selenium (Se)s and thus, contribute to population dynamics.Consumers are often overlooked as key drivers of vegetation structure and ecosystem functioning in coastal wetlands. This oversight is particularly apparent in Asia, where much of the variation in coastal wetland plant growth and composition is attributed to physical stress gradients. To address this knowledge gap and quantify the relative importance of consumers in Asian coastal wetlands across temporal variation in environmental stress, we conducted a two-year experiment spanning relatively spring wet (2018) and spring dry (2019) years in which we manipulated the presence of the numerically dominant herbivorous crab, Helice tientsinensis, and evaluated its effects on Phragmites australis growth and structure in a Yellow River Delta salt marsh. In spring wetter 2018, Phragmites biomass and stem density were 75% and 34% higher in Crab Exclusion relative to Ambient Crab plots. In 2019 which experienced spring drought and elevated soil salinity, Phragmites biomass and stem density remained similarly high relative to 2018 in Crab Exclusion plots, but fell further, to only 16% and 39% of levels of 2018 observed in Ambient Crab plots. Phragmites’ inflorescences density was also significantly reduced in Ambient Crab than Crab Exclusion plots in 2019. Together, these results highlight the significant role that crab herbivores can play in regulating Phragmites in Yellow River Delta salt marshes and suggest that the magnitude of their top-down control may be amplified, although in a non-additive manner, with spring drought stress in the region.Sediment quantity and quality in dam reservoirs is often neglected and usually only water quality is monitored in many countries, such as in Portugal. Nevertheless, there are risks associated to sedimentation in dam reservoirs, particularly considering that many dams in the world are ageing into an overextended lifetime. The present study was conducted with the goal to understand how sedimentation monitoring in Portuguese dam reservoirs could be attained, as that information is essential to tackle any sedimentation effects that may have occurred. A dam reservoir in the north of Portugal, the Venda Nova reservoir, was selected for the study. First, historical relevant data was compiled and then new data for sediment and water quality analysis was collected. The results show that since the reservoir was filled, after the dam construction conclusion in 1951, the reservoir morphology has been affected by the sedimentation at different rates and underwent multiple transformations from 1946 (before the dam construction) to 2004 (last topobathymetric survey). An expressive morphology transformation detected in 1984 led to an intervention to remove some of the accumulated material. It was verified, as well, that the asymmetrical contamination of the sediments and water is still strongly influenced by a nearby mine even if now inactive. The methodology applied, if periodically used, will be helpful to understand the sedimentation phenomena in Portuguese and other European dam reservoirs where reservoir sedimentation has been overlooked, and can play a key role to improve river continuity and to comply with the Environmental Quality Standards under the EU Water Framework Directive (2000/60/EC).Due to intensive marine activities and the use of low-quality fuel oils, the marine transport accounts for a considerable part of air pollution in the transportation sector. Although ships provide the convenient transportation, they exhaust a large number of hazardous pollutants, especially nitrogen oxides (NOx) and sulfur oxides (SOx), which have a significant side effect on environment and human health. To alleviate the impact of global shipping on the environment, international maritime organization established the more stringent emission regulations from Tier I to Tier III in order to reduce emissions from ships. As a result, various emission reduction technologies need to be developed in order to meet more stringent regulations in the future and reduce the pollutant emissions. Under this background, it is indispensable to examine the existing emission reduction technologies when exploring another possible method to reduce the pollutant emissions. Based on a significant number of related literatures, it is general to utilize the marine alternative fuels to reduce the pollutant emissions. Especially, liquid natural gas (LNG) is considered to be one of the most promising marine alternative fuels due to its economy and environmentally friendly features. This review thus aims to summarize the different emission reduction technologies of marine diesel engines through three reduction paths of fuel optimization, pre-combustion control and exhaust after-treatment. Furthermore, the utilization of LNG in the marine diesel engines are evaluated comprehensively from three aspects of environmental protection, energy structure and economic benefits. At the end, some suggestions on the future research are given based on the extensive review on the state-of-the-art literature.Afforestation is a significant cause of global peatland degradation. In some regions, afforested bogs are now undergoing clear-felling and restoration, often known as forest-to-bog restoration. We studied differences in water-table depth (WTD) and porewater chemistry between intact, afforested, and restored bogs at a raised bog and blanket bog location. Solute concentrations and principal component analysis suggested that water-table drawdown and higher electrical conductivity (EC) and ammonium (NH4-N) concentrations were associated with afforestation. In contrast, higher dissolved organic carbon (DOC) and phosphate (PO4-P) concentrations were associated with deforestation. Drying-rewetting cycles influenced seasonal variability in solute concentrations, particularly in shallower porewater at the raised bog location. WTD was significantly deeper in the oldest raised bog restoration site (~9 years post-restoration) than the intact bog (mean difference = 6.2 cm). However, WTD in the oldest blanket bog restoration site (~17 years post-restoration), where furrows had been blocked, was comparable to the intact bog (mean difference = 1.2 cm). When averaged for all porewater depths, NH4-N concentrations were significantly higher in the afforested than the intact sites (mean difference = 0.77 mg L-1) whereas significant differences between the oldest restoration sites and the intact sites included higher PO4-P (mean difference = 70 μg L-1) in the raised bog and higher DOC (mean difference = 5.6 mg L-1), EC (mean difference = 19 μS cm-1) and lower SUVA254 (mean difference = 0.13 L mg-1 m-1) in the blanket bog. Results indicate felled waste (brash) may be a significant source of soluble C and PO4-P. Mean porewater PO4-P concentrations were between two and five times higher in furrows and drains in which brash had accumulated compared to other locations in the same sites where brash had not accumulated. Creating and maintaining brash-free buffer zones may therefore minimise freshwater impacts.Increasing soil loss and the scarcity of useful land requires new reusing strategies. Thus, recovery of polluted soils recovery offers a chance for economic and social regeneration. With this objective, different soil cleaning technologies have been developed during the last few decades. On one hand, classical physical and/or chemical technologies can be found which are efficient, but have high costs and impacts upon ecosystems. On the other hand, biological methods (such as phytoremediation, bioremediation and vermiremediation) are relatively cost effective and eco-friendly, but also more time-consuming. These biological methods and their yields have been widely studied but little is known about the interaction between different soil cleaning methods. The combination of different biological strategies could lead to an improvement in remediation performance. Hence, in the present work, different micro-, vermi- and phyto-remediation combinations are applied in a sewage sludge polluted landfill in Gernika-Lumo tment, as highlighted by the battery of ecotoxicological tests and bioassays performed with earthworms, plants and bacteria.Sea purslane Halimione portulacoides (L.) Aellen is a candidate extractive species for coastal Integrated Multi-Trophic Aquaculture (IMTA) to recycle the dissolved inorganic nitrogen (DIN) and phosphorus (DIP) wasted by excretive species. To test its suitability, saline aquaculture effluents were simulated in the laboratory using a hydroponics approach to cultivate the plants. Nutrient extraction efficiency, growth performance and nutritional profile were assessed under a range of DIN and DIP concentrations representing three different aquaculture intensification regimes and using Hoagland’s solution as a control. Over a 10-week period, hydroponic units under non-limited N and P conditions displayed daily extraction rates between 1.5 and 2.8 mg DIN-N L-1 day-1 and 0.1-0.2 mg DIP-P L-1 day-1 and yielded between 63.0 and 73.0 g m-2 day-1 of H. portulacoides biomass. Relatively to biomass produced, H. portulacoides extracted between 2.6 and 4.2 mg DIN-N g-1 and 0.1-0.4 mg DIP-P g-1. The treatment with low-input of DIN and DIP (6.4 mg N L-1 and 0.7 mg P L-1) induced some degree of nutrient limitation, as suggested by the extremely high extraction efficiencies of DIN extraction (99%) in parallel with lower productivity. The nutritional profile of H. portulacoides leaves is comparable to that of other edible halophytes and leafy greens and could be a low-sodium alternative to salt in its lyophilized form. From the present study, we conclude that the edible halophyte H. portulacoides can be highly productive in hydroponics using saline water irrigation with non-limiting concentrations of DIN and DIP and is, therefore, a suitable extractive species for coastal IMTA in brackish waters.The habenula is among the evolutionarily most conserved parts of the brain and has been known for its role in the control of behavior to cope with aversive stimuli. Recent studies in zebrafish have revealed the novel roles of the two parallel neural pathways from the dorsal habenula to its target, the interpeduncular nucleus, in the control of behavioral choice whether to behave dominantly or submissively in the social conflict. They are modifiable depending on the internal state of the fish such as hunger and play another important role in orientation of attention whether to direct it internally to oneself or externally to others. These studies, therefore, are revealing a novel role for the habenula as the integrated switchboard for concertedly controlling behavior either as a winner with self-centered (idiothetic) attention or a loser with others-oriented (allothetic) attention.

For animals that live in social groups, the ability to recognize conspecifics is essential. Recent studies of both human patients and animal models have vigorously sought to discern the precise mechanisms by which hippocampal neurons and neural circuits contribute to the encoding, consolidation, storage, and retrieval of social memory. In particular, optogenetic manipulation enables us to investigate the presence of memory engrams.

We recently revealed the presence of social memory engrams in hippocampal ventral CA1 neurons, using optogenetic manipulation and calcium (Ca

) imaging.

In the present manuscript, we discuss the current viewpoints on two hippocampal subregions in regards to social memory representation, namely dorsal CA2 for information processing and ventral CA1 for the storage of social memory, specifically from the perspectives of behavioral neuroscience and neurophysiology.

In the present manuscript, we discuss the current viewpoints on two hippocampal subregions in regards to social memory representation, namely dorsal CA2 for information processing and ventral CA1 for the storage of social memory, specifically from the perspectives of behavioral neuroscience and neurophysiology.Water resources are of strategic importance for socioeconomic development. Many hydrological models (HMs) and land surface models (LSMs) have been developed for water resources assessment. However, systematic evaluation of discharge simulation from multiple models is still lacking in the Lancang-Mekong River basin. Here, we evaluated the performances of ten HMs and LSMs by evaluating their simulated discharge against observations at the basin scale. The selected models were within the Inter-Sectoral Impact Model Intercomparison Project (ISI-MIP2a) framework driven by Global Soil Wetness Project 3 (GSWP3) climate forcing data. Five discharge percentile series were used to evaluate the model performances for low, mean, and high flows. The intercomparison according to four statistical criteria revealed considerable differences exist in model performances for different discharge percentiles, indicating a large uncertainty caused by the choice of models with different degree of physical complexity and sensitivity to the quality of the input data. The models generally performed better for high flow than for low flow. Furthermore, the models generally performed better in downstream than in upstream, with the exception of close to the estuary, where complex processes involving interactions between freshwater and saline water are present. It is not surprising that the two calibrated model (WaterGAP2 and WAYS) are superior over the other models. This systematic intercomparison provides insights into the model behaviours and accuracies in discharges predicting with varying intensities, which can aid in quantifying uncertainties in water resources simulation at the basin scale.Ferroptosis is a form of cell death caused by iron-dependent lipid peroxidation. Cancer cells increase cystine uptake for the synthesis of glutathione (GSH), which is used by glutathione peroxidase 4 to reduce lipid peroxides. Here, we report that cystine deprivation in glioblastoma cells, but not inhibition of GSH synthesis by l-buthionine sulfoximine (BSO), induces ferroptosis. We found that cystine deprivation decreased the protein levels of ferritin heavy chain FTH1, whereas it was increased by BSO treatment. The lysosome inhibitor bafilomycin A1 or deletion of nuclear receptor coactivator 4 (NCOA4) inhibited cystine deprivation-induced decrease in FTH1 protein levels and cell death. In addition, cystine deprivation induced microtubule-associated protein light chain 3 (LC3)-II protein accumulation, suggesting that cystine deprivation induces ferritinophagy. BSO causes cell death when glioblastoma cells are treated with iron inducers, ferrous ammonium sulfate or hemin. On the other hand, cystine deprivation-induced degradation of FTH1 and cell death required glutamine. This study suggests that ferritinophagy, in addition to GSH depletion, plays an important role in cystine deprivation-induced ferroptosis in glioblastoma cells.CRISPR-Cas systems, including Cas9 and Cpf1 (Cas12a), are promising tools for generating gene knockout mouse models. Unlike Cas9, Cpf1 can generate multiple crRNAs from a single concatemeric crRNA precursor, which is favorable for multiplex gene editing. Recently, a hybrid guide RNA (hgRNA) system employing both Cas9 and Cpf1 was developed for multiplex gene editing. As the crRNA of Cpf1 was linked to the 3′ end of the sgRNA for Cas9, it can be split into separate guide RNAs by Cpf1. To examine whether this Cas9-Cpf1 hybrid system is suitable for multiplex gene knockouts in the mouse embryo, we generated an hgRNA that simultaneously targets the mouse Il10ra gene by Cas9 and mouse Dr3 (or Tnfrsf25, death receptor3) gene by Cpf1. The expression of hgRNA from a single promoter induced significant indels at each gene in cultured mouse cells upon the co-expression of both Cas9 and Cpf1. Interestingly, the hgRNA exhibited comparable Cas9-mediated indel activity without Cpf1 expression. Similarly, when the hgRNA was co-microinjected with both Cas9 and Cpf1 mRNAs into mouse zygotes at the pronuclear stage, founder mice were generated harboring mutations in both the Il10ra and Dr3 genes. However, when Cas9 mRNA was used alone without Cpf1 mRNA, the mouse Il10ra gene targeting was significantly decreased. These results indicate that the hgRNA system is a possible tool for multiplex gene targeting in the mouse embryo.In this report, we describe a truncated Deinococcus radiodurans 1-deoxy-D-xylulose-5-phosphate synthase (DXS) protein that retains enzymatic activity, while slowing protein degradation and showing improved crystallization properties. With modern drug-design approaches relying heavily on the elucidation of atomic interactions of potential new drugs with their targets, the need for co-crystal structures with the compounds of interest is high. DXS itself is a promising drug target, as it catalyzes the first reaction in the 2-C-methyl-D-erythritol 4-phosphate (MEP)-pathway for the biosynthesis of the universal precursors of terpenes, which are essential secondary metabolites. In contrast to many bacteria and pathogens, which employ the MEP pathway, mammals use the distinct mevalonate-pathway for the biosynthesis of these precursors, which makes all enzymes of the MEP-pathway potential new targets for the development of anti-infectives. However, crystallization of DXS has proven to be challenging while the first X-ray structures from Escherichia coli and D. radiodurans were solved in 2004, since then only two additions have been made in 2019 that were obtained under anoxic conditions. The presented site of truncation can potentially also be transferred to other homologues, opening up the possibility for the determination of crystal structures from pathogenic species, which until now could not be crystallized. This manuscript also provides a further example that truncation of a variable region of a protein can lead to improved structural data.

A growing number of epidemiological studies show associations between environmental factors and impaired cardiometabolic health. However, evidence is scarce concerning these risk factors and their impact on metabolic syndrome (MetS). This analysis aims to investigate associations between long-term exposure to air pollution, road traffic noise, residential greenness, and MetS.

We used data of the first (F4, 2006-2008) and second (FF4, 2013-2014) follow-up of the population-based KORA S4 survey in the region of Augsburg, Germany, to investigate associations between exposures and MetS prevalence at F4 (N=2883) and MetS incidence at FF4 (N=1192; average follow-up 6.5years). Residential long-term exposures to air pollution – including particulate matter (PM) with a diameter<10µm (PM

), PM<2.5µm (PM

), PM between 2.5 and 10µm (PM

), absorbance of PM

(PM2.5

), particle number concentration (PNC), nitrogen dioxide (NO

), ozone (O

) – and road traffic noise were modeled by land-use regression models eases in PM10 (OR 1.15; 95% confidence interval [95% CI] 1.02, 1.29), PM2.5 (OR 1.14; 95% CI 1.02, 1.28), PMcoarse (OR 1.14; 95% CI 1.02, 1.27), and PM2.5abs (OR 1.17; 95% CI 1.03, 1.32). Results further showed negative, but non-significant associations between exposure to greenness and prevalent and incident MetS. No effects were seen for exposure to road traffic noise. Joint Odds Ratios from multi-exposure models were higher than ORs from models with only one exposure.The ubiquitous use of organophosphate flame retardants and plasticizers (PFRs) in a variety of consumer products has led to widespread human exposure. Since certain PFRs are developmental and carcinogenic toxicants, detailed exposure assessments are essential to investigate the risk associated with environmental exposure levels. However, such data are still lacking for European countries. In this study, concentrations of thirteen PFR metabolites were measured in urine samples from 600 adolescents from Flanders, Belgium. 1-Hydroxy-2-propyl bis(1-chloro-2-propyl) phosphate (BCIPHIPP), diphenyl phosphate (DPHP), bis(1,3-dichloro-isopropyl) phosphate (BDCIPP), 2-hydroxyethyl bis(2-butoxyethyl) phosphate (BBOEHEP), 2-ethylhexyl phenyl phosphate (EHPHP) and 2-ethyl-5-hydroxyhexyl diphenyl phosphate (5-HO-EHDPHP) were frequently detected (>83%) in all participants. Comparisons with study populations from outside the EU showed that urinary levels of DPHP, BDCIPP and BCIPHIPP were generally within the same range. Only exposure to 2-ethylhexyl diphenyl phosphate (EHDPHP) was presumably higher in Flemish adolescents. However, determinants analysis through multivariate regression analyses did not reveal significant predictors that may explain this finding. Significantly higher levels of BDCIPP were observed in participants with new decorations at home, while adolescents with highly educated parents had higher levels of BBOEHEP and BDCIPP. Furthermore, multiple PFR metabolite concentrations followed a seasonal pattern. Estimated daily intakes (EDIs) were calculated from the internal dose by including fractions of urinary excretion (FUE) estimated in in vitro metabolism studies. EDIs ranged from 6.3 ng/kg bw/day for TBOEP to 567.7 ng/kg bw/day for EHDPHP, which were well below the available oral reference doses for all investigated PFRs. This suggests that the associated risk is low at present. This is the first report on internal exposure to seven commonly used PFRs in a European population.The brain comprises many different cell types with specialized functions which respond and adapt to the continuously changing environment, through tight spatiotemporal regulation of gene expression. The three-dimentional (3D) organisation of the genome is increasingly recognized as a major feature of gene regulation in brain cells, for the activation, repression and poising of gene expression, and in coupling transcription with RNA processing and transport. Here, we discuss the importance of dynamic chromatin organisation in the developmental patterning of the brain, and its role in fine tuning brain activity and plasticity. A better understanding of how disease-associated mutations interfere with chromatin organisation and long-range gene regulation will help reveal the molecular mechanisms underlying complex neurodevelopmental and neuropsychiatric disorders.An electrical immuno-sandwich assay utilizing an electrokinetic-based streaming current method for signal transduction is proposed. The method records the changes in streaming current, first when a target molecule binds to the capture probes immobilized on the inner surface of a silica micro-capillary, and then when the detection probes interact with the bound target molecules on the surface. The difference in signals in these two steps constitute the response of the assay, which offers better target selectivity and a linear concentration dependent response for a target concentration within the range 0.2-100 nM. The proof of concept is demonstrated by detecting different concentrations of Immunoglobulin G (IgG) in both phosphate buffered saline (PBS) and spiked in E. coli cell lysate. A superior target specificity for the sandwich assay compared to the corresponding direct assay is demonstrated along with a limit of detection of 90 pM in PBS. The prospect of improving the detection sensitivity was theoretically analysed, which indicated that the charge contrast between the target and the detection probe plays a crucial role in determining the signal. This aspect was then experimentally validated by modulating the zeta potential of the detection probe by conjugating negatively charged DNA oligonucleotides. The length of the conjugated DNA was varied from 5 to 30 nucleotides, altering the zeta potential of the detection probe from -9.3 ± 0.8 mV to -20.1 ± 0.9 mV. The measurements showed a clear and consistent enhancement of detection signal as a function of DNA lengths. The results presented here conclusively demonstrate the role of electric charge in detection sensitivity as well as the prospect for further improvement. The study therefore is a step forward in developing highly selective and sensitive electrokinetic assays for possible application in clinical investigations.The limited reaction time and sample volume in the confined space of microfluidic devices give considerable importance to the development of more effective biosensing interfaces. Herein, the self-assembling of tetrahedral framework nucleic acids (FNAs) with controllable size on the interface of the microfluidic microchannels is studied. Compared with macroscopic turbulence control on traditional micro-structured microfluidic surface, the novel FNA-engineered microfluidic interface successfully constructs a 3D reaction space at nanoscale by raising DNA probes away from the surface. This FNA interface dramatically improves the reaction kinetics during molecular recognition due to extremely ordered orientation, configuration and density of DNA probes on the surface. Finally, the FNA-engineered interface is applied in a novel multi-functional microfluidic platform, towards a „one-stop” assay of Escherichia coli O157 H7 (E. coli O157 H7), integrating capture, release, enrichment, cell culture and antimicrobial susceptibility testing (AST). With the FNA-aptamer probe, we achieved an enhanced bacterial detecting efficiency (10 CFU/mL) plus excellent selectivity and precision. The appicability was strongly demonstrated when the biosensor was successfully applied in real samples, including the analysis of antibiotic susceptibility and minimum inhibitory concentration (MIC) of E. coli O157 H7 among different antibiotics. The application of FNA interface will open a wide avenue for the development of microfluidic biosensors for other pathogenic microorganisms or circulating tumor cells (CTC) simply by changing the aptamers.Combining electrochemiluminescence (ECL) with nanozyme amplification provides unique advantages for the detection of antibiotic residues. Herein, a molecularly imprinted chloramphenicol (CAP) sensor was established based on aggregation-induced (AI)-ECL and nanozyme amplification. Covalent organic framework materials with AI-ECL groups (COF-AI-ECL) and nanozyme Co3O4 were synthesised as the signal element and the amplification element, respectively. Subsequently, using CAP as a template molecule, a molecularly imprinted polymer (MIP) was fabricated on the electrode surface modified with COF-AI-ECL and Co3O4. The ECL signal of COF-AI-ECL was catalytically amplified by Co3O4, whereas CAP effectively quenched this signal. Consequently, the ECL signal was controlled by the elution and adsorption of CAP by the MIP, thus establishing a new method for CAP detection. Unlike traditional ECL reagent, COF-AI-ECL exhibited a stable and strong ECL signal. Therefore, COF-AI-ECL in combination with the MIP provided greater sensitivity and enhanced selectivity. The linear range of the developed CAP sensor was 5 × 10-13 to 4 × 10-10 mol/L, with a detection limit of 1.18 × 10-13 mol/L. Moreover, the recoveries range of 85.0%-106.2% were obtained for the detection of CAP in real honey, milk, and chicken samples, indicating the potential of this sensor design for the detection of trace antibiotic residues in food safety applications.Super-resolution fluorescence microscopy has emerged as a powerful tool for studying mitochondrial dynamics in living cells. However, the lack of photostable and chemstable probe makes long-term super-resolution imaging of mitochondria still a challenging work. Herein, we reported a 4-azetidinyl-naphthliamide derived SNAP-tag probe AN-BG exhibiting excellent fluorogenicity and photostability for long-term super-resolution imaging of mitochondrial dynamics. The azetidinyl group and naphthalimide fluorophore are in a flat conformation which can effectively suppress twisted intramolecular charge transfer and then effectively improve the brightness and photostability. This planarized molecular structure is conducive to the formation of fluorescence-quenched J-aggregates, and the protein labeling process will depolymerize the probes and restore fluorescence. Fluorescent labeling mitochondrial inner membrane proteins via SNAP tags overcomes the shortcomings that variations in mitochondrial inner membrane potential will release probes attached to mitochondria by electrostatic interactions. Therefore, AN-BG realized the stable labeling of mitochondria and the long-term imaging of mitochondrial dynamics under super-resolution microscopy.Myocardial dysfunction caused by cardiomyocyte apoptosis under ischemic and hypoxic conditions is the pathological basis of most cardiovascular diseases. Current diagnosis of myocardial dysfunction still focuses on the symptomatic stage, usually after the occurrence of the irreversible remodelling and functional impairment. Thus, early stage identification of the apoptotic cardiomyocytes induced by hypoxia is highly significant for preventing the onset and delaying the progression of myocardial dysfunction. Herein, a novel Au-Se nanoprobe with strong anti-interference capability was developed for simultaneous real-time in situ monitoring the expression of Lon protease (Lon) and Caspase-3 with high-fidelity in living cardiomyocytes. As Lon upregulation plays a major role in the initiation of hypoxia-induced apoptosis and Caspase-3 is a marker protein for apoptosis, the nanoprobe has been successfully applied for imaging the activation of Lon-Caspase-3 apoptotic signalling pathway and assessing the state of cardiomyocytes under hypoxic conditions. Furthermore, combining with mitochondrial H2O2 probe-MitoPY1, the nanoprobe was also used to confirm the synergistic effect of Lon and ROS on hypoxia-induced apoptosis of cardiomyocytes and evaluate the function of ROS scavenger on attenuating such apoptosis. This work proposed a promising strategy for early diagnosis, prevention and treatment of hypoxic-ischemic myocardial dysfunction.A fusion enzyme composed of an Aspergillus flavus-derived flavin adenine dinucleotide glucose dehydrogenase (AfGDH) and an electron transfer domain of Phanerochaete chrysosporium-derived cellobiose dehydrogenase (Pcyb) was previously reported to show the direct electron transfer (DET) ability to an electrode. However, its slow intramolecular electron transfer (IET) rate from the FAD to the heme, limited the sensor signals. In this study, fusion FADGDH (Pcyb-AfGDH) enzymes were strategically redesigned by performing docking simulation, following surface-electrostatic potential estimation in the predicted area. Based on these predictions, we selected the amino acid substitution on Glu324, or on Asn408 to Lys to increase the positive charge at the rim of the interdomain region. Pcyb-AfGDH mutants were recombinantly produced using Pichia pastoris as the host microorganism, and their IET was evaluated. Spectroscopic observations showed that the Glu324Lys (E324K) and Asn408Lys (N408K) Pcyb-AfGDH mutants showed approximately 1.70- and 9.0-fold faster IET than that of wildtype Pcyb-AfGDH, respectively. Electrochemical evaluation revealed that the mutant Pcyb-AfGDH-immobilized electrodes showed higher DET current values than that of the wildtype Pcyb-AfGDH-immobilized electrodes at pH 6.5, which was approximately 9-fold higher in the E324K mutant and 15-fold higher in the N408K mutant, than in the wildtype. Glucose enzyme sensors employing N408K mutant was able to measure glucose concentration under physiological condition using artificial interstitial fluid at pH 7.4, whereas the one with wildtype Pcyb-AfGDH was not. These results indicated that the sensor employed the redesigned mutant Pcyb-AfGDH can be used for future continuous glucose monitoring system based on direct electron transfer principle. (247 words).

Knee osteoarthritis accounts for more years of disability than all other forms of osteoarthritis combined. Gait kinetic and kinematic changes, in addition to reduced gait speed, are commonly observed. This study investigates whether core activation, which modifies lower extremity movement in young, active populations, can alter the gait and baseline core activation of those with knee osteoarthritis as compared to controls, and alter osteoarthritic knee pain.

Forty-four participants (22 controls and 22 with knee osteoarthritis) underwent biomechanical gait assessment, examining kinetic and kinematic variables, in addition to gait speed, with and without volitional transversus abdominis activation. Surface electromyography was used to measure baseline transversus abdominis activation under both conditions. Knee pain ratings were examined for those with knee osteoarthritis.

No significant biomechanical differences were observed within groups, or in the time/group interaction. Between-groups kinetic (time to first peak ground reaction force and amplitude of second peak ground reaction force) and gait speed differences were observed under both conditions. There were no differences in baseline electromyography activation between or within-groups, or within-group for self-reported pain for the osteoarthritic group.

Although previous studies have shown the benefit of core activation in correcting lower extremity movement patterns and kinetic loading in young, athletic populations, this study is the first to show this is not the case for persons with knee osteoarthritis. Future studies should examine the value of a progressive core stabilization program, of sufficient dose and mode, in correcting the observed gait differences in those with knee osteoarthritis.

NCT03776981.

NCT03776981.

To assess the effects of the initial stepping limb on posterior fall recovery in individuals with chronic stroke, as well as to determine the benefits of fall-recovery training on these outcomes.

This was a single-group intervention study of 13 individuals with chronic stroke. Participants performed up to six training sessions, each including progressively challenging, treadmill-induced perturbations from a standing position. Progressions focused on initial steps with the paretic or non-paretic limb. The highest perturbation level achieved, the proportion of successful recoveries, step and trunk kinematics, as well as stance-limb muscle activation about the ankle were compared between the initial stepping limbs in the first session. Limb-specific outcomes were also compared between the first and last training sessions.

In the first session, initial steps with the non-paretic limb were associated with a higher proportion of success and larger perturbations than steps with the paretic limb (p=0.02, Cohen’s d=0.8). Paretic-limb steps were wider relative to the center of mass (CoM; p=0.01, d=1.3), likely due to an initial standing position with the CoM closer to the non-paretic limb (p=0.01, d=1.4). In the last training session, participants recovered from a higher proportion of perturbations and advanced to larger perturbations (p<0.05, d>0.6). There were no notable changes in kinematic or electromyography variables with training (p>0.07, d<0.5).

The skill of posterior stepping in response to a perturbation can be improved with practice in those with chronic stroke, we were not able to identify consistent underlying kinematic mechanisms behind this adaptation.

The skill of posterior stepping in response to a perturbation can be improved with practice in those with chronic stroke, we were not able to identify consistent underlying kinematic mechanisms behind this adaptation.The defining feature of the eukaryotic cell, the nucleus, is bounded by a double envelope. This envelope and the nuclear pores within it play a critical role in separating the genome from the cytoplasm. It also presents cells with a challenge. How are cells to remodel the nuclear compartment boundary during mitosis without compromising nuclear function? In the two billion years since the emergence of the first cells with a nucleus, eukaryotes have evolved a range of strategies to do this. At one extreme, the nucleus is disassembled upon entry into mitosis and then reassembled anew in the two daughter cells. At the other, cells maintain an intact nuclear compartment boundary throughout the division process. In this review, we discuss common features of the division process that underpin remodelling mechanisms, the topological challenges involved and speculate on the selective pressures that may drive the evolution of distinct modes of division.Glucagon-like peptide-1 receptor (GLP1R) is a seven-transmembrane-spanning helices membrane protein expressed in multiple human tissues including pancreatic islets, lung, brain, heart and central nervous system (CNS). GLP1R agonists are commonly used as antidiabetic drugs, but a neuroprotective function in neurodegenerative disorders is emerging. Here, we established two iPSC lines from a patient harboring a rare homozygous splice site variant in GLP1R (NM_002062.3; c.402 + 3delG). This patient displays severe developmental delay and epileptic encephalopathy. Therefore, the derivation of these iPSC lines constitutes a primary model to study the molecular pathology of GLP1R dysfunction and develop novel therapeutic targets.