Impact of Rhizosphere Biostimulation on Cd Transport and Isotope Fractionation in Cd-Tolerant and Hyperaccumulating Plants Based on MC-ICP-MS and NanoSIMS.

Phytoremediation efficiency can be enhanced by regulating rhizosphere processes, and the Cd isotope is a useful approach for deciphering Cd transport processes in soil-plant systems. However, the effects of adsorption and complexation on Cd isotope fractionation during the rhizosphere processes remain unclear. Here, we cultivated the Cd hyperaccumulator Sedum alfredii and Cd-tolerance Sedum spectabile in three different soils with citric acid applied as a degradable rhizosphere biostimulant. Cellular elemental distributions in the tissues and Cd isotope compositions were determined through NanoSIMS and MC-ICP-MS, respectively. Cd precipitation/adsorption on cell walls and intracellular regional distribution were the main mechanisms of Cd tolerance in S. spectabile. Plant roots became enriched with heavier Cd isotopes relative to the surrounding soils upon increasing secretion of rhizosphere organic acids. This indicates that organic matter with O and N functional groups preferentially chelates heavy Cd isotopes. In addition, Cd isotope fractionation between roots and shoots varies within the three soils, which could be due to the influence of protein and metallothionein contents in roots and leaves. The finding indicates that sulfur-containing ligands preferentially chelate light Cd isotopes. This study suggests that organic ligands play a vital role in Cd isotope fractionation and consequent hyperaccumulation of soil-plant systems.

Sources and health risks of heavy metals in kindergarten dust: The role of particle size.

Particle size effects significantly impact the concentration and toxicity of heavy metals (HMs) in dust. Nevertheless, the differences in concentrations, sources, and risks of HMs in dust with different particle sizes are unclear. Therefore, guided by the definition of atmospheric particulate matter, dust samples with particle sizes under 1000 µm (DT1000), 100 µm (DT100), and 63 µm (DT63) from Beijing kindergartens were collected. The concentrations of HMs (e.g., Cd, Pb, Zn, Ni, Cr, Ba, Cu, V, Mn, Co, and Ti) in dust samples with different particle sizes were measured. Besides, the differences in HM concentrations, contamination levels, sources, and source-oriented health risks in dust samples of different particle sizes were systematically explored. The results show that the concentrations of Mn, V, Zn, and Cd gradually increase with decreasing dust particle sizes, the concentrations of Ba and Pb show a decreasing trend, and the concentrations of Cr, Cu, Ni, and Co display an increasing and then decreasing trend. The degree of contamination of HMs in dust of different particle sizes varies, with Cd being the most dominant contaminant. Compared with DT1000 and DT63, DT100 is the most polluted. In addition, the sources of HMs in DT1000, DT100, and DT63 become more single with decreasing particle size, which may be mainly due to the particle-size effect inducing the redistribution of HMs in different sources. Notably, the potential health risk is higher in DT100 than in DT1000 and DT63. The highest contribution of industrial sources to the health risk is found in DT100, which is mainly caused by highly toxic chromium (Cr). This work emphasizes the importance of considering particle size in risk assessment and pollution control, which can provide a theoretical basis for precise management of HMs pollution in dust.

Insights into Health Risks of Face Paint Application to Opera Performers: The Release of Heavy Metals and Stage-Light-Induced Production of Reactive Oxygen Species.

Face paints used by opera performers have been shown to contain high levels of heavy metals. However, whether frequent exposure, via dermal contact and inadvertent oral ingestion, results in occupational diseases is unknown, as is the potential exacerbation of toxicity by high-intensity irradiation from stage lights. In this study, we examined the release of Cr, Cu, Pb, and Zn from 40 face paints and the consequent health risks posed by different practical scenarios involving their use. The results showed that the in vitro bioaccessibility (IVBA) of Cr, Cu, Pb, and Zn in the tested products was, on average, 7.0, 5.5, 19.9, and 7.9% through oral ingestion and 1.1, 2.2, 1.6, and 1.2% through dermal contact, respectively. Stage light irradiation significantly increased the IVBA associated with dermal contact, to the average of 4.8, 34.9, 5.7, and 1.9% for Cr, Cu, Pb, and Zn, respectively. The increase was mainly due to the light-induced generation of reactive oxygen species, particularly hydroxyl free radicals. The vitality and transcriptional response of 3D skin models as well as a quantitative risk assessment of skin sensitization indicated that dermal contact with face paints may induce predictable skin damage and potentially other skin diseases. Long-term exposure to face paints on stage may also pose a non-carcinogenic health risk. The demonstrated health risks to opera performers of face paint exposure should lead to strict regulations regarding the content of theatrical face paints.

Toxic effects of polystyrene nanoplastics and polybrominated diphenyl ethers to zebrafish (Danio rerio).

Nanoplastics (NPs) are good carriers of persistent organic pollutants (POPs) such as polybrominated diphenyl ethers (PBDEs), and can alter their bioavailability and toxic impacts to aquatic organisms. This study highlights the single and combined toxic effects of polystyrene nanoplastics (PS-NPs) and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47, one of the dominant congeners of PBDEs) on zebrafish embryos after an exposure duration of up to 120 hpf. Results showed that PS-NPs and BDE-47 co-exposure exacerbated the morphological deformities in terms of pericardial edema, yolk sac edema and curved tail in zebrafish larvae. Compared to BDE-47 single exposure, the combined exposure caused lower survival rates, shorter body lengths, and accelerated spontaneous movements. Further, PS-NPs were quickly aggregated on the surface of the embryonic chorions covered almost the entire membrane at 12 and 48 hpf, and concentration dependent accumulation was also found in the brain, mouth, trunk, gills, heart, liver and gastrointestinal tract at the larval stages. During the recovery period (7 days), PS-NPs were released from all the organs, with the highest elimination from the gastrointestinal tract. Histopathological examination revealed that co-exposure caused greater damage to retinal structures, muscle fibers and cartilage tissues. Responses of hypothalamic-pituitary-thyroid axis (CRH, TSHß, NIS, TTR, Dio2, TG, TRα and TRß) and reproduction (Esr2 and Vtg1) related genes were also investigated, and results showed that the co-exposure induced more significant upregulated expressions of TSHß, TG, Doi 2, and TRß, compared to BDE-47 single exposure. In conclusion, co-exposure to NPs and BDE-47 exacerbated developmental and thyroid toxicity in zebrafish, generally elucidating the toxicological effects mediated by complex chemical interactions between NPs with POPs in the freshwater environment.

Foliar Application of SiO2 Nanoparticles Alters Soil Metabolite Profiles and Microbial Community Composition in the Pakchoi (Brassica chinensis L.) Rhizosphere Grown in Contaminated Mine Soil.

Silica nanoparticles (SiO2-NPs) are promising in nanoenabled agriculture due to their large surface area and biocompatible properties. Understanding the fundamental interaction between SiO2-NPs and plants is important for their sustainable use. Here, 3 week-old pakchoi (Brassica chinensis L.) plants were sprayed with SiO2-NPs every 3 days for 15 days (5 mg of SiO2-NPs per plant), after which the phenotypes, biochemical properties, and molecular responses of the plants were evaluated. The changes in rhizosphere metabolites were characterized by gas chromatography-mass spectrometry (GC-MS)-based metabolomics, and the response of soil microorganisms to the SiO2-NPs were characterized by high-throughput bacterial 16S rRNA and fungal internal transcribed spacer (ITS) gene sequencing. The results showed that the SiO2-NP spray had no adverse effects on photosynthesis of pakchoi plants nor on their biomass. However, the rhizosphere metabolite profile was remarkably altered upon foliar exposure to SiO2-NPs. Significant increases in the relative abundance of several metabolites, including sugars and sugar alcohols (1.3-9.3-fold), fatty acids (1.5-18.0-fold), and small organic acids (1.5-66.9-fold), and significant decreases in the amino acid levels (60-100%) indicated the altered carbon and nitrogen pool in the rhizosphere. Although the community structure was unchanged, several bacterial (Rhodobacteraceae and Paenibacillus) and fungal (Chaetomium) genera in the rhizosphere involved in carbon and nitrogen cycles were increased. Our results provide novel insights into the environmental effects of SiO2-NPs and point out that foliar application of NPs can alter the soil metabolite profile.

Low Concentrations of Silver Nanoparticles and Silver Ions Perturb the Antioxidant Defense System and Nitrogen Metabolism in N2-Fixing Cyanobacteria.

Although toxic effects of silver nanoparticles (AgNPs) on aquatic organisms have been extensively reported, responses of nitrogen-fixing cyanobacteria to AgNPs/Ag+ under environmentally relevant concentrations are largely unknown. Here, cyanobacteria were exposed to different concentrations of AgNPs (0.01, 0.1, and 1 mg/L) or Ag+ (0.1, 1, and 10 µg/L) for 96 h. The impacts of AgNPs and Ag+ on photosynthesis and N2 fixation in cyanobacteria (Nostoc sphaeroides) were evaluated. In addition, gas chromatography-mass spectrometry (GC-MS)-based metabolomics was employed to give an instantaneous snapshot of the physiological status of the cells under AgNP/Ag+ exposure. Exposure to high doses of AgNPs (1 mg/L) or Ag+ (10 µg/L) caused growth inhibition, reactive oxygen species overproduction, malondialdehyde accumulation, and decreased N2 fixation. In contrast, low doses of AgNPs (0.01 and 0.1 mg/L) and Ag+ (0.1 and 1 µg/L) did not induce observable responses. However, metabolomics revealed that metabolic reprogramming occurred even at low concentrations of AgNP and Ag+ exposure. Levels of a number of antioxidant defense-related metabolites, especially phenolic acid and polyphenols (gallic acid, resveratrol, isochlorogenic acid, chlorogenic acid, cinnamic acid, 3-hydroxybenzoic acid, epicatechin, catechin, and ferulic acid), significantly decreased in response to AgNPs or Ag+. This indicates that AgNPs and Ag+ can disrupt the antioxidant defense system and disturb nitrogen metabolism even at low-dose exposure. Metabolomics was shown to be a powerful tool to detect "invisible" changes, not observable by typical phenotypic-based endpoints.

C60 Fullerols Enhance Copper Toxicity and Alter the Leaf Metabolite and Protein Profile in Cucumber.

Abiotic and biotic stress induce the production of reactive oxygen species (ROS), which limit crop production. Little is known about ROS reduction through the application of exogenous scavengers. In this study, C60 fullerol, a free radical scavenger, was foliar applied to three-week-old cucumber plants (1 or 2 mg/plant) before exposure to copper ions (5 mg/plant). Results showed that C60 fullerols augmented Cu toxicity by increasing the influx of Cu ions into cells (170% and 511%, respectively, for 1 and 2 mg of C60 fullerols/plant). We further use metabolomics and proteomics to investigate the mechanism of plant response to C60 fullerols. Metabolomics revealed that C60 fullerols up-regulated antioxidant metabolites including 3-hydroxyflavone, 1,2,4-benzenetriol, and methyl trans-cinnamate, among others, while it down-regulated cell membrane metabolites (linolenic and palmitoleic acid). Proteomics analysis revealed that C60 fullerols up-regulated chloroplast proteins involved in water photolysis (PSII protein), light-harvesting (CAB), ATP production (ATP synthase), pigment fixation (Mg-PPIX), and electron transport ( Cyt b6f). Chlorophyll fluorescence measurement showed that C60 fullerols significantly accelerated the electron transport rate in leaves (13.3% and 9.4%, respectively, for 1 and 2 mg C60 fullerols/plant). The global view of the metabolic pathway network suggests that C60 fullerols accelerated electron transport rate, which induced ROS overproduction in chloroplast thylakoids. Plant activated antioxidant and defense pathways to protect the cell from ROS damaging. The revealed benefit (enhance electron transport) and risk (alter membrane composition) suggest a cautious use of C60 fullerols for agricultural application.

Characteristics of cadmium accumulation and isotope fractionation in higher plants.

Cadmium (Cd) pollution of the soil is an important global environmental issue owing to its great toxicity. The study of metal isotope fractionation is a novel technique that could be used to identify and quantify metal uptake and transport mechanisms in plant. In this study, cadmium tolerant Ricinus communis and hyperaccumulator Solanum nigrum have been cultured in different Cd concentration nutrient solutions. The Cd isotope values, metal elements concentrations in the organs (root, stem and leaf) in the two plant species have been measured during the growth periods (10d, 15d, 20d, 25d, and 30d). The results indicate that the organs of S. nigrum could be enriched with lighter Cd isotopes compared with R. communis. In addition, the Cd isotope fractionation become smaller when the plants were subjected to high Cd toxicity, which indicates that Cd isotope fractionation reflected the extent of Cd toxicity to plants. This study advances our current view of Cd translocation machination in plants.

Multiple Sulfur Isotope Constraints on Sources and Formation Processes of Sulfate in Beijing PM2.5 Aerosol.

Recently air pollution is seriously threatening the health of millions of people in China. The multiple sulfur isotopic composition of sulfate in PM2.5 samples collected in Beijing is used to better constrain potential sources and formation processes of sulfate aerosol. The Δ33S values of sulfate in PM2.5 show a pronounced seasonality with positive values in spring, summer and autumn and negative values in winter. Positive Δ33S anomalies are interpreted to result from SO2 photolysis with self-shielding, and may reflect air mass transport between the troposphere and the stratosphere. The negative Δ33S signature (-0.300‰ < Δ33S < 0‰) in winter is possibly related to incomplete combustion of coal in residential stoves during the heating season, implying that sulfur dioxide released from residential stoves in more rural areas is an important contributor to atmospheric sulfate. However, negative Δ33S anomalies (-0.664‰ < Δ33S ← 0.300‰) in winter and positive Δ33S anomalies (0.300‰ < Δ33S < 0.480‰) in spring, summer, and autumn suggest sulfur isotopic equilibrium on an annual time frame, which may provide an implication for the absence of mass-independent fractionation of sulfur isotopes (S-MIF) in younger sediments. Results obtained here reveal that reducing the usage of coal and improving the heating system in rural areas will be important for efficiently decreasing the emissions of sulfur in China and beyond.

Interaction between sulfur and lead in toxicity, iron plaque formation and lead accumulation in rice plant.

Human activities have resulted in lead and sulfur accumulation in paddy soils in parts of southern China. A combined soil-sand pot experiment was conducted to investigate the influence of S supply on iron plaque formation and Pb accumulation in rice (Oryza sativa L.) under two Pb levels (0 and 600 mg kg(-1)), combined with four S concentrations (0, 30, 60, and 120 mg kg(-1)). Results showed that S supply significantly decreased Pb accumulation in straw and grains of rice. This result may be attributed to the enhancement of Fe plaque formation, decrease of Pb availability in soil, and increase of reduced glutathione (GSH) in rice leaves. Moderate S supply (30 mg kg(-1)) significantly increased Fe plaque formation on the root surface and in the rhizosphere, whereas excessive S supply (60 and 120 mg kg(-1)) significantly decreased the amounts of iron plaque on the root surface. Sulfur supply significantly enhanced the GSH contents in leaves of rice plants under Pb treatment. With excessive S application, the rice root acted as a more effective barrier to Pb accumulation compared with iron plaque. Excessive S supply may result in a higher monosulfide toxicity and decreased iron plaque formation on the root surface during flooded conditions. However, excessive S supply could effectively decrease Pb availability in soils and reduce Pb accumulation in rice plants.

Cadmium accumulation and tolerance of two castor cultivars in relation to antioxidant systems.

To investigate the effects of Cd on tolerance and antioxidant activities of castor, two different castor (Ricinus communis L.) cultivars (Zibo No. 5 and Zibo No. 8) were used for a hydroponic experiment (0, 1 and 2mg/L Cd) and a pot experiment using Cd-contaminated soil (34mg/kg) with the addition of ethylenedinitrilotetraacetic acid (EDTA). The results indicated that there were significant differences between the two cultivars with respect to Cd uptake in shoots (113-248mg/kg for Zibo No. 5 and 130-288mg/kg Zibo No. 8), biomass tolerance indexes (64.9%-74.6% for Zibo No. 5 and 80.1%-90.9% for Zibo No. 8) in the hydroponic experiment and survival rates (0% for Zibo No. 5 and 100% for Zibo No. 8) determined by the addition of EDTA in the pot experiment, suggesting that Zibo No. 8 has higher tolerance than Zibo No. 5. Moreover, the castor cultivars have low bioconcentration factors (4.80% for Zibo No. 5 and 5.43% for Zibo No. 8) and low translocation factors (<1%). Consequently, Zibo No. 8 can participate in Cd phytostabilization in highly Cd-polluted areas. The results indicated that glutathione (GSH) as a non-enzymatic antioxidant, and antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (GPX), were cultivar- and dose-dependent. The higher tolerance of Zibo No. 8 compared with Zibo No. 5 can be attributed to the higher GSH levels in the root and higher GPX activity in the leaf.

[Screening and engineering of a protocatechuic acid decarboxylase for efficient biosynthesis of catechol].

Catechol (CA) is an important chemical and pharmaceutical intermediate with wide applications. At present, CA is produced by phenol hydroxylation with non-renewable petrochemical resources, which causes serious environmental pollution. Hence, the biosynthesis of CA attracts much attention recently. However, due to the low activities of protocatechuic acid (PCA) decarboxylases, the production efficiency of biosynthetic catechol is too low to meet the requirements of large-scale industrial production. To improve the yield of CA, we screened 21 PCA decarboxylases from different species. RbAroY originated from Rikenellaceae had the best catalytic performance. The whole-cell biocatalyst ER11 with RbAroY was able to produce CA at a titer of 13.54 g/L. Then, the online tool HotSpot Wizard was employed to measure the enzyme stability, which revealed 10 potential mutation sites causing significant decreases in Gibbs free energy. The whole-cell biocatalyst ERT01 with the mutated RbAroYG99A could produce CA at a titer of 15.16 g/L, which increased by 12% compared with that of the wild-type whole-cell biocatalyst. After optimization of the biocatalytic conditions, the whole-cell biocatalyst ERT01 was able to produce CA at a titer of 25.70 g/L with PCA as the substrate. Finally, with the fermentation broth of 3-dehydroshikimate as the substrate, the whole-cell biocatalyst DER03 expressing both 3-dehydroshikimate dehydratase and PCA decarboxylase realized the production CA at a titer of 29.55 g/L, which is currently the highest biosynthetic titer reported. This study provides a reference for the industrial production of CA by biosynthesis.

Hazard profiles, distribution trends, and sources tracing of rare earth elements in dust of kindergartens in Beijing.

Children, the most vulnerable group in urban populations, are susceptible to the effects of pollution in urban environments. It is significant to evaluate the influence of rare earth elements (REEs) from kindergartens dust (KD) in Beijing on children's health. This study collected surface dust from 73 kindergartens in 16 districts of the mega-city of Beijing, and the concentrations of 14 REEs in KD, including La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu, were detected. The contamination levels, source apportionment, and health exposure risk of REEs were comprehensively investigated. The results indicate that the contamination levels of 14 REEs are within the acceptable range. Nevertheless, Eu, Ce, La, Pr, Nd, Gd, and Sm show high enrichment due to anthropogenic influence. Besides, KD is rich in light rare earth elements (LREEs) (90.97 mg kg-1) compared to heavy rare earth elements (HREEs) (8.65 mg kg-1). The distribution parameter patterns of REEs suggest that complicated anthropogenic sources influence the enrichment of REEs in KD. The main sources of REEs in KD include natural sources (40.64%), mixed high-tech industries and construction (33.89%), and mixed coal-fired, historical industrial, and transportation sources (26.47%). The primary pathway for daily intake of REEs in children is through ingestion, which presents a low but not negligible health risk. This study provides guidance for the effective risk management of REEs in KD.

Remarkable contamination characteristics, potential hazards and source apportionment of heavy metals in surface dust of kindergartens in a northern megacity of China.

It is essential to understand the impact of heavy metals (HMs) present in the surface dust (SD) of kindergartens on children, who are highly sensitive to contaminated dust in cities in their growth stage. A study was conducted on 11 types of HMs present in the SD of 73 kindergartens in Beijing. This study aims to assess the pollution levels and sources of eleven HMs in Beijing's kindergartens surface dust (KSD), and estimate the potential health risks in different populations and sources. The results indicate that Cd has the highest contamination in the KSD, followed by Pb, Zn, Ni, Ba, Cr, and Cu. The sources of these pollutants are identified as industrial sources (23.7%), natural sources (22.1%), traffic sources (30.4%), and construction sources (23.9%). Cancer risk is higher in children (4.02E-06) than in adults (8.93E-06). Notably, Cr is the priority pollutant in the KSD, and industrial and construction activities are the main sources of pollution that need to be controlled. The pollution in the central and surrounding areas is primarily caused by historical legacy industrial sites, transportation, urban development, and climate conditions. This work provides guidance to manage the pollution caused by HMs in the KSD of Beijing. ENVIRONMENTAL IMPLICATION: Children within urban populations are particularly sensitive to pollutants present in SD. Prolonged exposure to contaminated SD significantly heightens the likelihood of childhood illnesses. The pollution status and potential health risks of HMs within SD from urban kindergartens are comprehensively investigated. Additionally, the contributions from four primary sources are identified and quantified. Furthermore, a pollution-source-oriented assessment is adopted to clearly distinguish the diverse impacts of different sources on health risks, and the priority pollutants and sources are determined. This work holds pivotal importance for risk management, decision-making, and environmental control concerning HMs in KSD.

[Aspartate-ornithine granules in the treatment of nonalcoholic steatohepatitis: a multiple-dose parallel controlled clinical trial].

OBJECTIVE: To investigate the therapeutic efficacy and safety of aspartate-ornithine granules in patients with nonalcoholic steatohepatitis (NASH). METHODS: Seventy-two patients with NASH were included in this multiple-dose parallel controlled clinical trial and received a 12-week course of aspartate-ornithine granule treatment at either high-dose (6 g bid po; n = 38) or low-dose (3 g bid po; n = 34). Clinical efficacy was assessed by monitoring data from urinalysis, serologic tests (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and triglyceride (TG)), and abdominal computed tomography (CT) scan. Safety was assessed by occurrence of adverse events (fatigue, anorexia, abdominal distension, nausea, and vomiting). Statistical analyses were conducted to determine the significance of differences between parameters before (baseline) and after treatment. RESULTS: After 12 weeks of treatment, the liver and spleen CT ratios in both the high-dose group (0.89 +/- 0.19) and the low-dose group (0.80 +/- 0.15) were significantly higher than at baseline (S = 329, P less than 0.0001 and S = 246, P less than 0.0001); the overall improvement was more robust in the high-dose group (52.63%) than in the low-dose group (38.23%) (Z = -2.1042, P less than 0.05). After 6 and 12 weeks of treatment, the serum ALT levels in both the high-dose group and the low-dose group were significantly lower than at baseline (6 weeks: S = 324.5, P less than 0.0001 and S = 223, P less than 0.0001; 12 weeks: S = 370.5, P less than 0.0001 and S = 297.5, P less than 0.0001); the overall improvement was more robust in the high-dose group (79.0%) than in the low-dose group (53.0%) (Z = -2.0533, P less than 0.05). Similar trends were seen for the serum levels of AST and GGT after 6 and 12 weeks of treatment (all P less than 0.01) and serum levels of TG after 12 weeks of treatment. The rate of adverse reactions was low and similar between the two groups (high-dose: 4.8% and low-dose: 4.4%; all gastrointestinal). CONCLUSION: Aspartate-ornithine granule therapy was an effective and safe treatment of nonalcoholic steatohepatitis, with the higher dose of 6 g bid po providing more robust clinical benefit without affecting the safety profile.

Analysis of the Plasmid-Based ts-Mutant ΔfabA/pTS-fabA Reveals Its Lethality under Aerobic Growth Conditions That Is Suppressed by Mild Overexpression of desA at a Restrictive Temperature in Pseudomonas aeruginosa.

It is uncertain whether PA1610|fabA is essential or dispensable for growth on LB-agar plates under aerobic conditions in Pseudomonas aeruginosa PAO1. To examine its essentiality, we disrupted fabA in the presence of a native promoter-controlled complementary copy on ts-plasmid. In this analysis, we showed that the plasmid-based ts-mutant ΔfabA/pTS-fabA failed to grow at a restrictive temperature, consistent with the observation by Hoang and Schweizer (T. T. Hoang, H. P. Schweizer, J Bacteriol 179:5326-5332, 1997, https://doi.org/10.1128/jb.179.17.5326-5332.1997), and expanded on this by showing that ΔfabA exhibited curved cell morphology. On the other hand, strong induction of fabA-OE or PA3645|fabZ-OE impeded the growth of cells displaying oval morphology. Suppressor analysis revealed a mutant sup gene that suppressed a growth defect but not cell morphology of ΔfabA. Genome resequencing and transcriptomic profiling of sup identified PA0286|desA, whose promoter carried a single-nucleotide polymorphism (SNP), and transcription was significantly upregulated (level increase of >2-fold, P < 0.05). By integration of the SNP-bearing promoter-controlled desA gene into the chromosome of ΔfabA/pTS-fabA, we showed that the SNP is sufficient for ΔfabA to phenocopy the sup mutant. Furthermore, mild induction of the araC-PBAD-controlled desA gene but not desB rescued ΔfabA. These results validated that mild overexpression of desA fully suppressed the lethality but not the curved cell morphology of ΔfabA. Similarly, Zhu et al. (Zhu K, Choi K-H, Schweizer HP, Rock CO, Zhang Y-M, Mol Microbiol 60:260-273, 2006, https://doi.org/10.1111/j.1365-2958.2006.05088.x) showed that multicopy desA partially alleviated the slow growth phenotype of ΔfabA, the difference in which was that ΔfabA was viable. Taken together, our results demonstrate that fabA is essential for aerobic growth. We propose that the plasmid-based ts-allele is useful for exploring the genetic suppression interaction of essential genes of interest in P. aeruginosa. IMPORTANCE Pseudomonas aeruginosa is an opportunistic pathogen whose multidrug resistance demands new drug development. Fatty acids are essential for viability, and essential genes are ideal drug targets. However, the growth defect of essential gene mutants can be suppressed. Suppressors tend to be accumulated during the construction of essential gene deletion mutants, hampering the genetic analysis. To circumvent this issue, we constructed a deletion allele of fabA in the presence of a native promoter-controlled complementary copy in the ts-plasmid. In this analysis, we showed that ΔfabA/pTS-fabA failed to grow at a restrictive temperature, supporting its essentiality. Suppressor analysis revealed desA, whose promoter carried a SNP and whose transcription was upregulated. We validated that both the SNP-bearing promoter-controlled and regulable PBAD promoter-controlled desA suppressed the lethality of ΔfabA. Together, our results demonstrate that fabA is essential for aerobic growth. We propose that plasmid-based ts-alleles are suitable for genetic analysis of essential genes of interest.

Effects of non-invasive brain stimulation (NIBS) for executive function on subjects with ADHD: a protocol for a systematic review and meta-analysis.

INTRODUCTION: Attention-deficit/hyperactivity disorder (ADHD) is a prevalent neurodevelopmental disorder with a high risk of multiple mental health and social difficulties. Executive function domains are associated with distinct ADHD symptom burdens. Non-invasive brain stimulation (NIBS) mainly includes repetitive transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS), which is a promising technique, but its impact on the executive function of ADHD is uncertain. Therefore, the aim of this systematic review and meta-analysis is to derive solid and updated estimates on the effect of NIBS on executive function in children/adults with ADHD. METHODS AND ANALYSIS: A systematic search will be performed through EMBASE, MEDLINE, PsycINFO and Web of Science databases from inception until 22 August 2022. Handsearching of grey literature and the reference lists of selected articles will also be conducted. Empirical studies assessing the effect of NIBS (TMS or tDCS) on executive function in children or adults with ADHD will be included. Two investigators will independently perform literature identification, data extraction and risk of bias assessment. Relevant data will be pooled by a fixed-effects or random-effects model according to I2 statistic. Sensitivity analysis will be performed to test the robustness of the pooled estimates. Subgroup analyses will be conducted to investigate the potential heterogeneity. This protocol will generate a systematic review and meta-analysis that comprehensively synthesises the evidence on the NIBS treatment of executive function deficit of ADHD.Ethics approval is not required as this is a protocol for a systematic review of published literature. The results will be submitted to a peer-reviewed journal or a conference. PROSPERO REGISTRATION NUMBER: CRD42022356476.

Variations and its driven factors of anthropogenic nitrogen loads in the Yangtze River Economic Belt during 2000-2019.

Since the millennium, China has economically taken off with rapid urbanization, and anthropogenic nitrogen emission intensity has undergone remarkable changes. To better understand the impact of urbanization on anthropogenic nitrogen, this study calculated the spatio-temporal heterogeneity of anthropogenic nitrogen in the Yangtze River Economic Belt (YREB) since 2000, based on the estimation, using obstacle analysis to quantify the driving of industry and agriculture on N growth and using the gray model to analyze the impact of urbanization on N changes. Additionally, using the environmental pressure model to predict the future N load. The results indicated N load in the YREB increased rapidly from 21.4 Tg in 2001 to a peak of 24.5 Tg in 2012 and then decreased to 22.2 Tg in 2019. Although N flux gradually increased from the west to the east of the YREB, the growth rate had an opposite trend with a negative growth in the eastern region. Hotspots are mainly concentrated in urban agglomerations, which contributed to ~ 60% N load of the YREB, and the YREB contributed to ~ 90% N load of the Yangtze River Basin. Obstacle degree scores indicated wastewater was a major industrial driver of N growth before 2010, and then became waste gas; increased mechanization and fertilizer control effectively reduced nitrogen emissions during agricultural development. The gray analysis of urbanization indicated urban population, industry, and services had the strongest correlation with N load changes. Scenario simulations suggest N loads of the YREB remain at a high level by 2030; however, there are still opportunities to effectively control N growth through high technological innovation and reducing the proportion of industry under an enormous population. This research contributes to a better understanding of the impact of urbanization on anthropogenic nitrogen and helps developing countries to precisely control nitrogen hotspots and sources.

Neuro- and hepato-toxicity of polystyrene nanoplastics and polybrominated diphenyl ethers on early life stages of zebrafish.

Nanoplastics (NPs) are good carriers of persistent organic pollutants (POPs) such as polybrominated diphenyl ethers (PBDEs) and can modify their bioavailability and toxicity to aquatic organisms. This study highlights the single and combined toxic effects of polystyrene nanoplastics (PS-NPs) and 2,2 ',4,4 '-tetrabromodiphenyl ether (BDE-47, one of the major PBDE congeners) on zebrafish embryos after an exposure of up to 120 hpf. Our results showed that PS-NPs and BDE-47 formed larger particle aggregates during co-exposure, which attached to the surface of the yolk membrane and even changed its structure, and these particles also bioaccumulated in the intestine of zebrafish larvae, compared with the PS-NPs single exposure. Further, the co-exposure significantly increased mortality, accelerated voluntary movements, enhanced hatching rate, and decreased heart rate. Hepatoxicity analyses revealed that the mixture exposure induced a darker/browner liver colour, atrophied liver and greater hepatotoxicity in zebrafish larvae. In addition to increased ROS accumulation, the reduced expression of the antioxidant gpx1a gene and increased expression of cyp1a1 were found after co-treatment. Moreover, ache and chrn7α genes associated with neurocentral development, were significantly downregulated, mainly in the co-exposure group. In conclusion, simultaneous exposure to PS-NPs and BDE-47 exacerbated oxidative stress, developmental impacts, hepatotoxicity, and neurodevelopmental toxicity in zebrafish larvae. Therefore, neurotoxic effects of complex chemical interactions between PS-NPs and persistent organic pollutants in freshwater environments should be paid more attention.

Source partitioning using phosphate oxygen isotopes and multiple models in a large catchment.

Excessive phosphorus (P) loadings cause major pollution concerns in large catchments. Quantifying the point and nonpoint P sources of large catchments is essential for catchment P management. Although phosphate oxygen isotopes (δ18O(PO4)) can reveal P sources and P cycling in catchments, quantifying multiple P sources in a whole catchment should be a research focus. Therefore, this study aimed to quantitatively identify the proportions of multiple potential end members in a typical large catchment (the Yangtze River Catchment) by combining the phosphate oxygen isotopes, land use type, mixed end-element model, and a Bayesian model. The δ18O(PO4) values of river water varied spatially from 4.9‰ to18.3‰ in the wet season and 6.0‰ to 20.9‰ in the dry season. Minor seasonal differences but obvious spatial changes in δ18O(PO4) values could illustrate how human activity changed the functioning of the system. The results of isotopic mass balance and the Bayesian model confirmed that controlling agricultural P from fertilizers was the key to achieving P emission reduction goals by reducing P inputs. Additionally, the effective rural domestic sewage treatment, development of composting technology, and resource utilization of phosphogypsum waste could also contribute to catchment P control. P sources in catchment ecosystems can be assessed by coupling an isotope approach and multiple-models.