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Heterologous expression of mlrA in a photoautotrophic host – Engineering cyanobacteria to degrade microcystins
2018
Dexter, Jason | Dziga, Dariusz | Lv, Jing | Zhu, Junqi | Strzalka, Wojciech | Maksylewicz, Anna | Maroszek, Magdalena | Marek, Sylwia | Fu, Pengcheng
In this report, we establish proof-of-principle demonstrating for the first time genetic engineering of a photoautotrophic microorganism for bioremediation of naturally occurring cyanotoxins. In model cyanobacterium Synechocystis sp. PCC 6803 we have heterologously expressed Sphingopyxis sp. USTB-05 microcystinase (MlrA) bearing a 23 amino acid N-terminus secretion peptide from native Synechocystis sp. PCC 6803 PilA (sll1694). The resultant whole cell biocatalyst displayed about 3 times higher activity against microcystin-LR compared to a native MlrA host (Sphingomonas sp. ACM 3962), normalized for optical density. In addition, MlrA activity was found to be almost entirely located in the cyanobacterial cytosolic fraction, despite the presence of the secretion tag, with crude cellular extracts showing MlrA activity comparable to extracts from MlrA expressing E. coli. Furthermore, despite approximately 9.4-fold higher initial MlrA activity of a whole cell E. coli biocatalyst, utilization of a photoautotrophic chassis resulted in prolonged stability of MlrA activity when cultured under semi-natural conditions (using lake water), with the heterologous MlrA biocatalytic activity of the E. coli culture disappearing after 4 days, while the cyanobacterial host displayed activity (3% of initial activity) after 9 days. In addition, the cyanobacterial cell density was maintained over the duration of this experiment while the cell density of the E. coli culture rapidly declined. Lastly, failure to establish a stable cyanobacterial isolate expressing native MlrA (without the N-terminus tag) via the strong cpcB560 promoter draws attention to the use of peptide tags to positively modulate expression of potentially toxic proteins.
Mostrar más [+] Menos [-]Prenatal exposure to polychlorinated biphenyl and umbilical cord hormones and birth outcomes in an island population
2018
Tang, Mengling | Yin, Shanshan | Zhang, Jianyun | Chen, Kun | Jin, Mingjuan | Liu, Weiping
Prenatal exposure to polychlorinated biphenyl (PCB) is suspected to interfere with fetal development including reproductive and thyroid function and birth outcomes, but published evidence are still sparse. We designed a cross-sectional study to analyze the associations between PCB levels in umbilical cord sera and hormones and birth outcomes of mothers and newborns who are residents from an island. Seven indicator-PCB (PCB-28, 52, 101, 118, 138, 153, 180), and five reproductive hormones including luteotropic hormones (LH), estradiol (E2), testosterone (T), follicle-stimulating hormones (FSH) and anti-Mullerian hormones (AMH), and three thyroid hormones including tri-iodothyronine (T3), tetra-iodothyronine (T4) and thyroid stimulating hormones (TSH) were measured in 106 cord sera specimens. Birth outcomes include birth weight, length, head circumference, and gestational age. Multiple linear regression and quartile regression were used to analyze the associations between PCB and each of the hormones and birth outcomes, adjusting for selected potential confounders. The median value of total PCB in umbilical cord sera was 2.02 μg L−1 (IQR, 1.13–4.64). Several negative associations between PCB exposure and reproductive hormones were found. Among them, the β value of PCB-101 for FSH reached −0.38 (95%CI, −0.69, −0.07; p = 0.02). Moreover, we also found some sex-specific associations i.e. PCB-28 was negatively correlated with LH and T and PCB-118 was negatively correlated with T in male newborns but not in female newborns. The associations between PCB and birth outcomes seem to differ by molecular weight of the PCB congeners i.e. the low-chlorinated PCB congeners were negatively associated with gestational age and head circumference while high-chlorinated PCB congeners were positively associated birth weight and gestational age. In this study, we found that PCB congeners with different molecular weight has different associations with hormones and birth outcomes, and future studies are recommended to investigate underlying mechanisms of these associations.
Mostrar más [+] Menos [-]Microplastic and mesoplastic pollution in farmland soils in suburbs of Shanghai, China
2018
Liu, Mengting | Lu, Shibo | Song, Yang | Lei, Lili | Hu, Jiani | Lv, Weiwei | Zhou, Wenzong | Cao, Chengjin | Shi, Huahong | Yang, Xiaofeng | He, Defu
Microplastics are emerging pollutants which have been extensively detected in water environments. However, little is known about microplastic pollution in soil environments. In this study, we investigated microplastics and mesoplastics in farmland soils from twenty vegetable fields around the suburbs of Shanghai. In each site, three duplicate soil samples were collected from shallow (0–3 cm) and deep soils (3–6 cm), respectively. Microplastics (sizes of 20 μm - 5 mm) and mesoplastics (5 mm - 2 cm) were detected using methods of density extraction, 30% H₂O₂ digestion and micro-fourier transform infrared spectroscopy. The abundance of microplastics was 78.00 ± 12.91 and 62.50 ± 12.97 items kg⁻¹ in shallow and deep soils, respectively. While, mesoplastics were found with abundance of 6.75 ± 1.51 and 3.25 ± 1.04 items kg⁻¹ in shallow and deep soils. Among these micro(meso)plastics, 48.79% and 59.81% were in size of <1 mm in shallow and deep soils. The main morphotypes of microplastics included fiber, fragment and film, mostly in color of black or transparent. Moreover, we found that topsoil contained higher concentrations and larger sizes of micro(meso)plastics than deep soil. In addition, the vast majority of micro(meso)plastics were polypropylene (50.51%) and polyethylene (43.43%). This study reveals occurrence and characteristics of microplastic pollution in typical farmland soils. It provides important data for subsequent research on microplatics in the terrestrial ecosystem.
Mostrar más [+] Menos [-]Reconstructing the deposition environment and long-term fate of Chernobyl 137Cs at the floodplain scale through mobile gamma spectrometry
2018
Varley, Adam | Tyler, Andrew | Bondar, Yuri | Hosseini, Ali | Zabrotski, Viachaslau | Dowdall, Mark
Cs-137 is considered to be the most significant anthropogenic contributor to human dose and presents a particularly difficult remediation challenge after a dispersal following nuclear incident. The Chernobyl Nuclear Power Plant meltdown in April 1986 represents the largest nuclear accident in history and released over 80 PBq of 137Cs into the environment. As a result, much of the land in close proximity to Chernobyl, which includes the Polessie State Radioecology Reserve in Belarus, remains highly contaminated with 137Cs to such an extent they remain uninhabitable. Whilst there is a broad scale understanding of the depositional patterns within and beyond the exclusion zone, detailed mapping of the distribution is often limited. New developments in mobile gamma spectrometry provide the opportunity to map the fallout of 137Cs and begin to reconstruct the depositional environment and the long-term behaviour of 137Cs in the environment. Here, full gamma spectrum analysis using algorithms based on the peak-valley ratio derived from Monte Carlo simulations are used to estimate the total 137Cs deposition and its depth distribution in the soil. The results revealed a pattern of 137Cs distribution consistent with the deposition occurring at a time of flooding, which is validated by review of satellite imagery acquired at similar times of the year. The results were also consistent with systematic burial of the fallout 137Cs by annual flooding events. These results were validated by sediment cores collected along a transect across the flood plain. The true merit of the approach was confirmed by exposing new insights into the spatial distribution and long term fate of 137Cs across the floodplain. Such systematic patterns of behaviour are likely to be fundamental to the understanding of the radioecological behaviour of 137Cs whilst also providing a tracer for quantifying the ecological controls on sediment movement and deposition at a landscape scale.
Mostrar más [+] Menos [-]A metabolomics strategy to assess the combined toxicity of polycyclic aromatic hydrocarbons (PAHs) and short-chain chlorinated paraffins (SCCPs)
2018
Wang, Feidi | Zhang, Haijun | Geng, Ningbo | Ren, Xiaoqian | Zhang, Baoqin | Gong, Yufeng | Chen, Jiping
The combined toxicity of mixed chemicals is usually evaluated according to several specific endpoints, and other potentially toxic effects are disregarded. In this study, we provided a metabolomics strategy to achieve a comprehensive understanding of toxicological interactions between mixed chemicals on metabolism. The metabolic changes were quantified by a pseudotargeted analysis, and the types of combined effects were quantitatively discriminated according to the calculation of metabolic effect level index (MELI). The metabolomics strategy was used to assess the combined effects of polycyclic aromatic hydrocarbons (PAHs) and short-chain chlorinated paraffins (SCCPs) on the metabolism of human hepatoma HepG2 cells. Our data suggested that exposure to a combination of PAHs and SCCPs at human internal exposure levels could result in an additive effect on the overall metabolism, whereas diverse joint effects were observed on various metabolic pathways. The combined exposure could induce a synergistic up-regulation of phospholipid metabolism, an additive up-regulation of fatty acid metabolism, an additive down-regulation of tricarboxylic acid cycle and glycolysis, and an antagonistic effect on purine metabolism. SCCPs in the mixture acted as the primary driver for the acceleration of phospholipid and fatty acid metabolism. Lipid metabolism disorder caused by exposure to a combination of PAHs and SCCPs should be an important concern for human health.
Mostrar más [+] Menos [-]Toxicological effects on earthworms (Eisenia fetida) exposed to sub-lethal concentrations of BDE-47 and BDE-209 from a metabolic point
2018
Liang, Ruoyu | Chen, Juan | Shi, Yajuan | Lü, Yonglong | Sarvajayakesavalu, Suriyanarayanan | Xu, Xiangbo | Zheng, Xiaoqi | Kifāyatullāh, K̲h̲ān | Su, Chao
Earthworms improve the soil fertility and they are also sensitive to soil contaminants. Earthworms (Eisenia fetida), standard reference species, were usually chosen to culture and handle for toxicity tests. Metabolic responses in earthworms exposed to 2, 2′, 4, 4′-tetrabromodiphenyl ether (BDE-47) and decabromodiphenyl ether (BDE-209) were inhibitory and interfered with basal metabolism. In this study, 1H-NMR based metabolomics was used to identify sensitive biomarkers and explore metabolic responses of earthworms under sub-lethal BDE-47 and BDE-209 concentrations for 14 days. The results revealed that lactate was accumulated in earthworms exposed to BDE-47 and BDE-209. Glutamate increased significantly when the concentration of BDE-47 and BDE-209 reached 10 mg/kg. The BDE-47 exposure above 50 mg/kg concentration decreased the content of fumarate significantly, which was noticed different from that of BDE-209. Whereas, the BDE-207 or BDE-209 exposure increased the protein degradation into amino acids in vivo. The increased betaine content indicated that earthworms may maintain the cell osmotic pressure and protected enzyme activity by metabolic regulation. Moreover, the BDE-47 and BDE-209 exposure at 10 mg/kg changed most of the metabolites significantly, indicating that the metabolic responses were more sensitive than growth inhibition and gene expression. The metabolomics results revealed the toxic modes of BDE-47 and BDE-209 act on the osmoregulation, energy metabolism, nerve activities, tricarboxylic acid cycle and amino acids metabolism. Furthermore, our results highlighted that the 1H-NMR based metabolomics is a strong tool for identifying sensitive biomarkers and eco-toxicological assessment.
Mostrar más [+] Menos [-]Long-term exposure to ambient particulate matter (PM2.5) is associated with platelet counts in adults
2018
Zhang, Zilong | Chan, Ta-Chien | Guo, Cui | Chang, Ly-yun | Lin, Changqing | Chuang, Yuan Chieh | Jiang, Wun Kai | Ho, Kin Fai | Tam, Tony | Woo, Kam S. | Lau, Alexis K.H. | Lao, Xiang Qian
The prothrombotic effects of particulate matter (PM) may underlie the association of air pollution with increased risks of cardiovascular disease. This study aimed to investigate the association between long-term exposure to PM with an aerodynamic diameter ≤2.5 μm (PM2.5) and platelet counts, a marker of coagulation profiles.The study participants were from a cohort consisting of 362,396 Taiwanese adults who participated in a standard medical examination program between 2001 and 2014. Platelet counts were measured through Complete Blood Count tests. A satellite-based spatio-temporal model was used to estimate 2-year average ambient PM2.5 concentration at each participant's address. Mixed-effects linear regression models were used to investigate the association between PM2.5 exposure and platelet counts.This analysis included 175,959 men with 396,248 observations and 186,437 women with 397,877 observations. Every 10-μg/m3 increment in the 2-year average PM2.5 was associated with increases of 0.42% (95% CI: 0.38%, 0.47%) and 0.49% (95% CI: 0.44%, 0.54%) in platelet counts in men and women, respectively. A series of sensitivity analyses, including an analysis in participants free of cardiometabolic disorders, confirmed the robustness of the observed associations. Baseline data analyses showed that every 10-μg/m3 increment in PM2.5 was associated with higher risk of 17% and 14% of having elevated platelet counts (≥90th percentile) in men and women, respectively.Long-term exposure to PM2.5 appears to be associated with increased platelet counts, indicating potential adverse effects on blood coagulability.
Mostrar más [+] Menos [-]Comparative toxicity and bioaccumulation of two dinotefuran metabolites, UF and DN, in earthworms (Eisenia fetida)
2018
Liu, Tong | Zhang, Xiaolian | Wang, Xiuguo | Chen, Dan | Li, Yiqiang | Wang, Fenglong
Dinotefuran, as the latest generation of neonicotinoid insecticide, has broad application prospects around the world. However, dinotefuran is easily metabolized and the two main metabolites are 1-methyl-3-(tetrahydro-3-furylmethyl) urea (UF) and 1-methyl-3-(tetrahydro-3-furylmethyl) guanidium dihydrogen (DN). In the present study, the risks of UF and DN in soil on typical non-target species earthworm were investigated. In the same time, the degradation in soil and accumulation in earthworms of UF and DN were monitored. The present results showed that the toxicities of UF and DN were similar in their effect on earthworms and they were supertoxic pollutants to earthworms. The degradation trend in soil and accumulation trend in earthworms of UF and DN were consistent throughout the whole exposure period. At 1.0 mg/kg and 2.0 mg/kg, UF and DN could induce the excess production of ROS, resulting in oxidative stress effects in earthworm cells. The excess ROS induce changes in antioxidant enzyme activities, damage in biomacromolecules, and abnormal expression of function genes. The present results showed that UF and DN may have high risks for earthworms.
Mostrar más [+] Menos [-]Linking sub-individual and supra-individual effects in Daphnia magna exposed to sub-lethal concentration of chlorpyrifos
2018
Ferrario, Claudia | Parolini, Marco | De Felice, Beatrice | Villa, Sara | Finizio, Antonio
The main objective of the present study was to investigate possible links between sub-individual and supra-individual levels (i.e. population level) biomarkers in D. magna exposed to sublethal concentrations of the insecticide chlorpyrifos (CPF). To achieve the aim, 8-day old individuals were exposed for 96 h to two environmentally relevant concentrations of CPF (50 and 250 ng/L). Sub-individual level effects were investigated by measuring the activity of antioxidant (SOD, CAT, and GPx) and detoxifying (GST) enzymes, as well as by measuring the acetylcholinesterase (AChE) inhibition. In addition, the effects at supra-individual level were assessed by using a video-tracking system and analyzing changes in swimming capabilities (i.e. percentage of activity time, distance moved, and velocity). Our data have shown that daphnids exposed to both CPF concentrations were in a condition of stress which was highlighted by changes in both sub- and supra-individual biomarkers. Moreover, our results highlighted that the lowest tested CPF concentration did not modulate the antioxidant and detoxifying enzymes, whereas, an inhibition of AChE and a decrease of some parameters related to swimming behaviour (distance moved and velocity) were noted. On the contrary, significant changes in all the sub-individual biomarkers were measured at the highest tested concentration. In addition, organisms recovered the movement capability (distance moved) and also activate a mechanism of avoidance (increased swimming velocity). On the other hand, a reduction in the percent of active time was measured and this was attributed to the energy spent by organisms to activate antioxidant and detoxifying enzymes and the mechanism of avoidance. Based on these results, our study suggests the existence of a link between sub- and supra-individual levels, as the activation or non-activation in the antioxidant and detoxifying enzymes activities can led to different modifications of the swimming behaviour in D. magna.
Mostrar más [+] Menos [-]RETRACTED: Signalling cross-talk between nitric oxide and active oxygen in Trifolium repens L. plants responses to cadmium stress
2018
Liu, Shiliang | Yang, Rongjie | Tripathi, Durgesh Kumar | Li, Xi | Jiang, Mingyan | Lv, Bingyang | Ma, Mingdong | Chen, Qibing
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal).This article has been retracted at the request of the Editors-in-Chief.The authors have plagiarized part of a paper that had appeared in Plant, Cell & Environment, 29 (2006) 1532–1544. https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1365-3040.2006.01531.x The images that were reused were: Figures 1e, 1f, 1g, 1h, 2a, 2b, 2e, 2f.One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.
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