Unraveling the binding behavior of the antifouling biocide 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one with human serum albumin: Multi-spectroscopic, atomic force microscope, computational simulation, and esterase activity.

As a marine antifouling biocide, 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) exhibited high toxicity to marine organisms. This study investigated the interaction between DCOIT and human serum albumin (HSA) using several spectroscopic techniques combined with computer prediction methods. The UV-vis absorption spectra, Stern-Volmer constant (KSV) and fluorescence resonance energy transfer (FRET) results indicated that DCOIT caused static quenching of HSA fluorescence. The ΔG°, ΔH° and ΔS° values were -31.03 ± 0.17 kJ·mol-1, -133.54 ± 0.88 kJ·mol-1 and -348.46 ± 2.86 J.mol-1·K-1, respectively, suggesting that van der Waals forces and hydrogen bonds governed the spontaneous formation of the complex. Synchronous fluorescence and circular dichroism (CD) spectroscopy observed the burial of Trp residues within HSA and the unfolding of HSA secondary structure induced by DCOIT. Three-dimensional (3D) fluorescence and Atomic Force Microscopy (AFM) further detected DCOIT-induced loosening of HSA peptide chain structure. Site displacement experiments indicated that DCOIT binding at site I of HSA. Computational predictions indicated that hydrophobic interactions were also essential in the complex. The increased RMSD, Rg, SASA, and RMSF confirmed that DCOIT weakened the stability and compactness of HSA, rendering residues more flexible. Lastly, esterase activity assays demonstrated that DCOIT inhibited esterase activity and interfered with the human detoxification process.

Photocatalytic degradation of sulfonamides in suspensions of coral-like graphene carbon nitride with nitrogen vacancies.

Sulfonamides (SNs) belong to a category of broad-spectrum antibiotics, which have attracted growing concerns owing to the adverse effects on ecosystem. In this paper, coral-like graphitic carbon nitrides with nitrogen vacancies were prepared by polymerization of melamine in the presence of NH4Cl, and the effect of NH4Cl amount on the structure and photocatalytic performance of g-C3N4 in degradation of sulfonamide antibiotics such as sulfamethoxazole (SMX), sulfadiazine (SDZ) and sulfathiazole (STZ) was systematically studied. It was found that the addition of NH4Cl results in the formation of coral-like g-C3N4 with nitrogen vacancies, and optimal photocatalyst (PCN-1 sample) prepared with a melamine to NH4Cl mass ratio of 1:1 showed the highest photocatalytic activity towards SNs degradation due to the quick electron-hole migration, efficient separation capacity and excellent photoelectric properties. The electron paramagnetic resonance (EPR) technique was used to determine the reactive oxygen species (ROSs) that are responsible for the degradation of SNs, and the detailed degradation pathway of STZ was proposed according to the identification of the intermediates by liguid chromatography-high resolution mass spectrometry (LC-HRMS).

Functional polymer brushes for anti-microplastic pollution.

Image 1.

Effects of switching redox conditions on sediment phosphorus immobilization by calcium/aluminum composite capping: Performance, ecological safety and mechanisms.

There many materials were used in lake restoration to immobilize phosphorus (P) and reduce the effect of eutrophication. Among them, calcium/aluminum composite (CAC) showed a good capacity of P adsorption. However, a comprehensive of its performance, ecological safety, and the mechanism of P passivation in the aluminum-bound P (Al -P) dominated sediments under varying redox conditions remains incomplete. In the current study, both unwashed CAC (UCAC) and washed CAC (WCAC) showed good P adsorption properties, and the greatest maximum capacity for P adsorption (Qmax) reached 206.8 mg/g at pH 8.5 for UCAC. The SRP and TP in the overlying water of the uncapped sediments showed a decrease-increase-decrease trend in a sequence of transition from aerobic to anaerobic to re-aerobic stages. In contrast, the SRP and TP of the two CACs-capped sediments were maintained low. Phosphorus forms in the uncapped sediment also underwent significant changes during continuous variation of dissolved oxygen (DO) levels. In particular, the decrease in iron-bound P (Fe-P) and Al-P was significantly promoted in the anaerobic phase, and the released P was reabsorbed to form mainly Fe-P in the re-aerobic phase. The CACs-capping promoted the transformation of Fe-P to residual P (Res-P), forming a thick static layer in the surface sediment, thus significantly inhibiting sediment P release. Moreover, the CACs-capping did not induce the Al3+ leaching and significant changes of the microbial community in sediments, and their performances of P immobilization could keep stable to resist the redox variation, which promised to be a good choice for P passivation in eutrophic lake sediments dominated by Al/Fe-P. These findings also confirmed that the risk of P release from Al/Fe-P (mainly Al-P)-dominated sediments was strongly influenced by continuously changing redox conditions, and was probably enhanced by the formation of Fe-P from the resorption of the released P.

Research progress on eco-friendly superhydrophobic materials in environment, energy and biology.

In the past few years, bioinspired eco-friendly superhydrophobic materials (EFSMs) have made great breakthroughs, especially in the fields of environment, energy and biology, which have made remarkable contributions to the sustainable development of the natural environment. However, some potential challenges still exist, which urgently need to be systematically summarized to guide the future development of this field. Herein, in this review, initially, we discuss the five typical superhydrophobic models, namely, the Wenzel, Cassie, Wenzel-Cassie, "lotus", and "gecko" models. Then, the existence of superhydrophobic creatures in nature and artificial EFSMs are summarized. Then, we focus on the applications of EFSMs in the fields of environment (self-cleaning, wastewater purification, and membrane distillation), energy (solar evaporation, heat accumulation, and batteries), and biology (biosensors, biomedicine, antibacterial, and food packaging). Finally, the challenges and developments of eco-friendly superhydrophobic materials are highlighted.

Quantitative analysis of glutathione and cysteine S-conjugates of microcystin-LR in the liver, kidney and muscle of common carp (Cyprinus carpio) in Lake Taihu.

Tissue distribution of microcystin (MC)-LR-GSH, MC-LR-Cys and MC-LR of omnivorous fish in Lake Taihu was investigated. MC-LR and MC-LR-Cys were detected in liver, kidney and muscle. The concentration of MC-LR in liver and kidney was 0.052 µg g-1 DW and 0.067 µg g-1 DW, respectively. MC-LR-Cys appeared to be an important metabolite with average contents of 1.104 µg g-1 DW and 0.724 µg g-1 DW in liver and kidney, and the MC-LR-Cys/MC-LR ratio in liver and kidney reaching as high as 21.4 and 10.8. High MC-LR-Cys/MC-LR ratio and a significant correlation between MC-LR-Cys and MC-LR concentration in liver, suggest that liver is more active in detoxification of MC-LR by formation of MC-LR-Cys for omnivorous fish. Furthermore, there might be a balance between the accumulation and depuration/metabolism of MC-LR-Cys in kidney. The MC-LR-Cys can be formed in kidney directly, or transported from liver or other tissues, while the MC-LR-Cys in kidney might be dissociated to MC-LR or excreted. Although MC-LR and its metabolites were scarcely detected in muscle, it is necessary to investigate the distribution of toxic metabolites in edible muscle.

The temporal distribution, source and potential toxicity of polycyclic aromatic hydrocarbons in a sediment core from an urban lake in Wuhan, China.

The urban lakes in China, especially those with relatively small areas and closed watersheds, have suffered from serious pollution and are at a high risk of eutrophication due to the small amount of concern given to them. The temporal distribution profile, source apportionment and potential toxicity of parent polycyclic aromatic hydrocarbons (PAHs) and alkylated PAHs (APAHs) in a sediment core from an urban shallow lake with the above specifications, in the middle and lower reaches of the Yangtze River, China, were investigated in the present study. Parent PAHs, especially 4-6 ring ones, were dominant in the sediment core. The vertical profiles of 16 anthropogenic US Environmental Protection Agency (EPA) priority PAHs increased from a depth of 50 cm to 30 cm, then slightly decreased to the surface. However, biogenic perylene content decreased from bottom to surface. Most of the APAHs increased in concentration from bottom to surface. Source apportionment of EPA PAHs indicated the pyrogenic source was dominant in the sediment core. The domestic combustion of bituminous coal, coal tar and biomass can be identified as the main factors responsible for this. Toxicity assessment of selected parent PAHs demonstrated that benzo(a)pyrene accounted for the largest part of the toxic equivalent quantity (TEQ), and PAHs in the studied area possibly imposed adverse biological effects, especially for benthic organisms. Therefore, action should be taken to prevent an increase in the contamination from PAHs in the studied area, as well as in similar urban lakes in China, based on the specificity of individual watersheds.

Rapid conversion and reversible conjugation of glutathione detoxification of microcystins in bighead carp (Aristichthys nobilis).

The glutathione and cysteine conjugates of microcystin (MC-GSH and MC-Cys, respectively) are two important metabolites in the detoxification of microcystins (MCs). Although studies have quantitated both conjugates, the reason why the amounts of MC-GSH are much lower than those of MC-Cys in various animal organs remains unknown. In this study, MC-RR-GSH and MC-RR-Cys were respectively i.p. injected into the cyanobacteria-eating bighead carp (Aristichthys nobilis), to explore the biotransformation and detoxification mechanisms of the two conjugates. The contents of MC-RR, MC-RR-GSH, MC-RR-Cys and MC-RR-N-acetyl-cysteine (MC-RR-Nac, the acetylation product of MC-RR-Cys) in the liver, kidney, intestine and blood of bighead carp in both groups were quantified via liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). In the MC-RR-GSH-treated group, the MC-RR-Cys content in the kidney increased 96.7-fold from 0.25 to 0.5h post-injection, demonstrating that MC-RR-GSH acts as a highly reactive intermediate and is rapidly converted to MC-RR-Cys. The presence of MC-RR in both MC-RR-GSH- and MC-RR-Cys-treated groups indicates, for the first time, that MC conjugation with the thiol of GSH/Cys is a reversible process in vivo. Total MC-RR concentrations dissociated from MC-RR-Cys were lower than those from MC-RR-GSH, suggesting that MC-RR-Cys is more capable of detoxifying MC-RR. MC-RR-Cys was the most effectively excreted form in both the kidney and intestine, as the ratios of MC-RR-Cys to MC-RR reached as high as 15.2, 2.9 in the MC-RR-GSH-treated group and 63.4, 19.1 in the MC-RR-Cys-treated group. Whereas MC-RR-Nac could not be found in all of the samples of the present study. Our results indicate that MC-RR-GSH was rapidly converted to MC-RR-Cys and then excreted, and that both glutathione and cysteine conjugates could release MC-RR. This study quantitatively proves the importance of the GSH detoxification pathway and furthers our understanding of the biochemical mechanism by which bighead carp are resistant to toxic cyanobacteria.

Why mammals more susceptible to the hepatotoxic microcystins than fish: evidences from plasma and albumin protein binding through equilibrium dialysis.

To elucidate the interspecies variation of susceptibility to microcystins (MCs), fresh plasma and purified albumin from six kinds of mammals and fish were used in toxins-substances binding test. Protein contents in the test plasma were analyzed and the binding characteristics to MCs were compared. Two kinds of widely observed MCs, microcystin-LR (MC-LR) and microcystin-RR (MC-RR) were tested and data were collected through the method of equilibrium dialysis. It was found that total plasma protein and albumin content in mammals were nearly two times and four times higher than that in fish, respectively. In the test range of 0-100 µg/mL, binding rates of fish plasma to MCs were considered significant lower (p < 0.01) than that of mammals. And human plasma demonstrated the highest binding rate in mammals. In all the test species, plasma protein binding rates of MC-RR were significantly higher than MC-LR (p < 0.01). Besides, binding profiles of albumin were acquired under the protein content of 0.67 mg/mL. Human serum albumin demonstrated the highest affinity to MCs throughout the six species and differences among the other five species were considered not significant (p > 0.05). From the view of protein binding, it is concluded that both the variation of plasma protein composition and albumin binding characteristic could influence the existing form of MCs in circulation, change MCs utilization, alter MCs half-life and further contribute to the difference of susceptibility between mammals and fish.

Preliminary study of the distribution and accumulation of GSH/Cys metabolites of hepatotoxic microcystins-RR in common carp from a lake with protracted cyanobacterial bloom (Lake Taihu, China).

Tissue distributions and seasonal dynamics of glutathione and cysteine S-conjugates of microcystin-RR in feral fish from Lake Taihu were studied. High MC-RR-Cys was found in tissues, Mean MC-RR-Cys in kidney (0.253 µg g(-1) DW) was 4 times of that in liver (0.063 µg g(-1) DW). Ratios of MC-RR-Cys/MC-RR in liver/kidney reached as high as 5.3/39.8, respectively, meanwhile, kidney showed low accumulation of MC-RR and higher formation efficiency of MC-RR-Cys than liver (7.51× to liver), this suggested that MC-RR-Cys were significantly accumulated with the depletion of MC-RR, and it was selectively biotransformed to MC-RR-Cys in kidney for further excretion.

Metabolic response to oral microcystin-LR exposure in the rat by NMR-based metabonomic study.

Microcystin-LR (MCLR), a potent hepatotoxin, is causing increased risks to public health. Although the liver is the main target organ of MCLR, the metabolic profiling of liver in response to MCLR in vivo remains unknown. Here, we comprehensively analyzed the metabolic change of liver and ileal flushes in rat orally gavaged with MCLR by 1H nuclear magnetic resonance (NMR). Quantification of hepatic MCLR and its glutathione and cysteine conjugates by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was conducted. Metabonomics results revealed significant associations of MCLR-induced disruption of hepatic metabolisms with inhibition of nutrient absorption, as evidenced by a severe decrease of 12 amino acids in the liver and their corresponding elevation in ileal flushes. The hepatic metabolism signature of MCLR was characterized by significant inhibition of tyrosine anabolism and catabolism, three disrupted pathways of choline metabolism, glutathione exhaustion, and disturbed nucleotide synthesis. Notably, substantial alterations of hepatic metabolism were observable even at the low MCLR-treated group (0.04 mg/kg MCLR), although no apparent histological changes in liver were observed in the low- and medium-dosed groups. These observations offered novel insights into the microcystin hepatotoxic mechanism at a functional level, thereby facilitating further assessment and clarification of human health risk from MCs exposure.

Quantitatively evaluating detoxification of the hepatotoxic microcystins through the glutathione and cysteine pathway in the cyanobacteria-eating bighead carp.

Glutathione (GSH) and cysteine (Cys) conjugation have long been recognized to be important in the detoxification of microcystins (MCs) in animal organs, however, studies quantitatively estimating this process are rare, especially those simultaneously determining multiple toxins and their metabolites. This paper, for the first time, simultaneously quantified MC-LR (leucine arginine), MC-RR (arginine arginine), MCLR-GSH/Cys and MCRR-GSH/Cys in the liver, kidney, intestine and muscle of the cyanobacteria-eating bighead carp i.p. injected with two doses of MCs using liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). MCLR-Cys and MCRR-Cys content were much higher in kidney than in liver, intestine and muscle, suggesting the organotropism to kidney, while MCLR-GSH and MCRR-GSH were always below the detection limit. Bighead carp effectively metabolized MC-LR and MC-RR into the cysteine conjugates in kidney, as the ratios of MCLR-Cys to MC-LR and MCRR-Cys to MC-RR reached as high as 9.04 and 19.10, respectively. MC-LR and MC-RR were excreted mostly in the form of MCLR/RR-Cys rather than MCLR/RR-GSH, while MCs-GSH might act as mid-metabolites and changed to the more stable MCs-Cys rapidly. Cysteine conjugation of MCs appears to be an important biochemical mechanism for the cyanobacteria-eating fish to resist toxic cyanobacteria. A comparison of such detoxification mechanisms between fish and mammals would be interesting in the future studies.

Use of a generalized additive model to investigate key abiotic factors affecting microcystin cellular quotas in heavy bloom areas of Lake Taihu.

Lake Taihu is the third largest freshwater lake in China and is suffering from serious cyanobacterial blooms with the associated drinking water contamination by microcystin (MC) for millions of citizens. So far, most studies on MCs have been limited to two small bays, while systematic research on the whole lake is lacking. To explain the variations in MC concentrations during cyanobacterial bloom, a large-scale survey at 30 sites across the lake was conducted monthly in 2008. The health risks of MC exposure were high, especially in the northern area. Both Microcystis abundance and MC cellular quotas presented positive correlations with MC concentration in the bloom seasons, suggesting that the toxic risks during Microcystis proliferations were affected by variations in both Microcystis density and MC production per Microcystis cell. Use of a powerful predictive modeling tool named generalized additive model (GAM) helped visualize significant effects of abiotic factors related to carbon fixation and proliferation of Microcystis (conductivity, dissolved inorganic carbon (DIC), water temperature and pH) on MC cellular quotas from recruitment period of Microcystis to the bloom seasons, suggesting the possible use of these factors, in addition to Microcystis abundance, as warning signs to predict toxic events in the future. The interesting relationship between macrophytes and MC cellular quotas of Microcystis (i.e., high MC cellular quotas in the presence of macrophytes) needs further investigation.

Protein expression profiling in the zebrafish (Danio rerio) embryos exposed to the microcystin-LR.

Microcystin-leucine-arginine (MCLR) is the most toxic and the most commonly encountered variant of microcystins (MCs) in aquatic environment, and it has the potential for developmental toxicity. A number of previous studies have described the developing toxicity of MCLR based on conventional toxicological indices. However, the molecular mechanisms by which it expresses its toxicity during the early development remain largely unknown. To further our understanding of mechanisms of action and identify the potential protein biomarkers for MCLR exposure, a proteomic analysis was performed on developing zebrafish embryos exposed to 0.5 mg/L MCLR until 96 hours post-fertilization. 2-DE combined with MS was employed to detect and identify the protein profiles. Results showed that 75 spots from the 0.5 mg/L MCLR condition showed a significant increase or decrease in abundance compared with the control. In total, 40 proteins were identified. These proteins were mainly included in process related to oxidative stress, energetic metabolism, and the cytoskeleton assembly. MCLR exposure also affects the expression of the subunits of protein phosphatases 2A. Furthermore, the proteomic and transcriptional analysis of nine proteins was determined by Western blot and quantitative real-time PCR due to their correlation with the known MCLR toxic mechanisms. The consistent and discrepant results between protein and mRNA levels indicated complicated regulatory mechanisms of gene expression in response to MCLR exposure.

Factors affecting temporal and spatial variations of microcystins in Gonghu Bay of Lake Taihu, with potential risk of microcystin contamination to human health.

A field survey of the seasonal variation of microcystin (MC) concentration was performed in Gonghu Bay (a total of 15 sampling sites) of Lake Taihu from January to December 2008. Microcystis spp. biomass and intra-/extracellular MCs were significantly correlated with water temperature, suggesting the importance of temperature in cyanobacterial blooming in the lake. Higher MC concentration was found in summer and autumn, and peaks of Microcystis biomass and intra-/extracellular MC concentrations were all present in October. Spatially, risk of MCs was higher in littoral zones than in the pelagic area. There were significant correlations between N or P concentrations, and Microcystis biomass or MC content, suggesting that N and P levels affected MC production through influencing Microcystis biomass. Intra-/extracellular MCs and Microcystis biomass had negative exponential relationships with TN:TP, and the maximum values all occurred when TN:TP was <25. Multivariate analyses by PCCA indicated that intra- and extracellular MC concentrations had better correlations with biological factors (such as Microcystis biomass and chl-a) than with physicochemical factors. The maximum MC concentration reached up to 17 micrograms/L MC-LReq, considerably higher than the drinking water safety standard (1 micrograms/L) recommended by the WHO. So it is necessary to take measures to reduce the exposure risk of cyanobacterial toxins to human beings.

Development and validation of a liquid chromatography-tandem mass spectrometry assay for the simultaneous quantitation of microcystin-RR and its metabolites in fish liver.

A novel method for identification and quantification of microcystin-RR (MC-RR) and its metabolites (MC-RR-GSH and MC-RR-Cys) in the fish liver was developed and validated. These analytes were simultaneously extracted from fish liver using water containing EDTA with 5% acetic acid, followed by a mixed-mode cation-exchange SPE (Oasis MCX) and subsequently determined by liquid chromatography-electrospray ionization ion trap mass spectrometry (LC-ESI-ITMS). Extraction parameters including volume and pH of eluting solvents, were optimized. Best recoveries were obtained by using 10 mL of 15% ammonia solution in methanol. The mean recoveries at three concentrations (0.2, 1.0, and 5.0 microg g(-1) dry weight [DW]) for MC-RR, MC-RR-GSH and MC-RR-Cys were 93.6-99%, 68.1-73.6% and 90.0-95.2%, respectively. Method detection limit (MDL) were 4, 7 and 5 ng g(-1) DW for MC-RR, MC-RR-GSH and MC-RR-Cys, respectively. Limits of quantification (LOQs) for MC-RR, MC-RR-GSH and MC-RR-Cys were calculated to be 10, 18 and 13 ng g(-1) DW, respectively. Finally, this method was successfully applied to the identification and quantification of MC-RR, MC-RR-GSH and MC-RR-Cys in the liver of bighead carp with acute exposure of MCs.