Combining Network Pharmacology with Molecular Docking for Mechanistic Research on Thyroid Dysfunction Caused by Polybrominated Diphenyl Ethers and Their Metabolites.

Network pharmacology was used to illuminate the targets and pathways of polybrominated diphenyl ethers (PBDEs) causing thyroid dysfunction. A protein-protein interaction (PPI) network was constructed. Molecular docking was applied to analyze PBDEs and key targets according to the network pharmacology results. A total of 247 targets were found to be related to 16 PBDEs. Ten key targets with direct action were identified, including the top five PIK3R1, MAPK1, SRC, RXRA, and TP53. Gene Ontology (GO) functional enrichment analysis identified 75 biological items. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified 62 pathways mainly related to the regulation of the thyroid hormone signaling pathway, MAPK signaling pathway, PI3K-Akt signaling, pathways in cancer, proteoglycans in cancer, progesterone-mediated oocyte maturation, and others. The molecular docking results showed that BDE-99, BDE-153, 5-OH-BDE47, 5'-OH-BDE99, 5-BDE47 sulfate, and 5'-BDE99 sulfate have a good binding effect with the kernel targets. PBDEs could interfere with the thyroid hormone endocrine through multiple targets and biological pathways, and metabolites demonstrated stronger effects than the prototypes. This research provides a basis for further research on the toxicological effects and molecular mechanisms of PBDEs and their metabolites. Furthermore, the application of network pharmacology to the study of the toxicity mechanisms of environmental pollutants provides a new methodology for environmental toxicology.

In-vitro assessment of the binding mechanism of oxyfluorfen (herbicide) with garlic phytocystatin: multi-spectroscopic and isothermal titration calorimetric study.

Oxyfluorfen (2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene) is a nitrophenyl ether herbicide. Phytocystatins are crucial plant proteins which regulate various physiological processes and are also responsible for maintaining protease-antiprotease balance within plants. Thus, the present article deciphers the interaction of oxyfluorfen with garlic phytocystatin (GPC) through various spectroscopic and calorimetric techniques. The cysteine proteinase inhibitory assay was done to assess the inhibitory action of GPC in the presence of oxyfluorfen. The GPC loses its inhibitory activity in the presence of oxyfluorfen. The complex formation of GPC-oxyfluorfen was shown by UV absorption spectroscopy. The intrinsic fluorescence experiment affirmed the quenching of GPC in the presence of oxyfluorfen. The Stern-Volmer quenching constant and binding constant was obtained as 6.89 × 103 M-1 and 9.72 × 103 M-1, respectively. Synchronous fluorescence showed the alteration in the microenvironment around tyrosine residues. 3D fluorescence suggested the perturbation in the polarity around aromatic residues. The isothermal titration experiment suggests that the interaction of oxyfluorfen with GPC is a thermodynamically favorable reaction. Secondary structure alteration of GPC in the presence of oxyfluorfen was studied by circular dichroism (CD). The CD result showed a reduction in the α-helical content of GPC on interaction with oxyfluorfen. Consequently, all these outcomes affirmed the formation of GPC-oxyfluorfen complex along with the structural and conformational alteration. This study identifies and signifies that the exposure of oxyfluorfen induces stress within the plant system. Communicated by Ramaswamy H. Sarma.

Microbial degradation of high impact polystyrene (HIPS), an e-plastic with decabromodiphenyl oxide and antimony trioxide.

Accumulation of electronic waste has increased catastrophically and out of that various plastic resins constitute one of the leading thrown out materials in the electronic machinery. Enrichment medium, containing high impact polystyrene (HIPS) with decabromodiphenyl oxide and antimony trioxide as sole carbon source, was used to isolate microbial cultures. The viability of these cultures in the e-plastic containing mineral medium was further confirmed by triphenyl tetrazolium chloride (TTC) reduction test. Four cultures were identified by 16S rRNA sequencing as Enterobacter sp., Citrobacter sedlakii, Alcaligenes sp. and Brevundimonas diminuta. Biodegradation experiments were carried out in flask level and gelatin supplementation (0.1% w/v) along with HIPS had increased the degradation rate to a maximum of 12.4% (w/w) within 30days. This is the first report for this kind of material. The comparison of FTIR, NMR, and TGA analysis of original and degraded e-plastic films revealed structural changes under microbial treatment. Polystyrene degradation intermediates in the culture supernatant were also detected using HPLC analysis. The gravity of biodegradation was validated by morphological changes under scanning electron microscope. All isolates displayed depolymerase activity to substantiate enzymatic degradation of e-plastic.

Influences of artificial root exudate components on the behaviors of BDE-28 and BDE-47 in soils: desorption, availability, and biodegradation.

Behaviors of BDE-28 and BDE-47 in two distinct soils (Phaeozem and Acrisol) as affected by the separate addition of root exudate components (i.e., oxalic acid, glycine, and fructose) were investigated by a soil microcosm incubation experiment. The results showed that root exudate components promoted the desorption of BDE-28 (57.6-235.0 %) and BDE-47 (56.9-223.7 %) from the soils due to the enhancement of their water solubilities. The addition of root exudate components increased the n-butanol extractability of BDE-28 and BDE-47 by 20.3-72.5 and 48.6-169.2 %, respectively, which had a positive correlation with the concentrations of dissolved organic carbon (DOC) in the soils (p < 0.01), suggesting that the increase of DOC in the soils by root exudate components was the major factor to enhance the extractability. Fructose and oxalic acid promoted the desorption and increased the availability of BDE-28 and BDE-47 in the soils more efficiently than glycine. The addition of different root exudate components resulted in distinct shifts in soil microbial community structure (p < 0.05). Oxalic acid caused the greatest impacts on the soil bacterial communities and increased the degradation rates of BDE-28 and BDE-47 most obviously. The findings of this study clarified the roles of root exudate components in affecting the behaviors of polybrominated diphenyl ethers (PBDEs) in soils.

Enhanced soil washing process for the remediation of PBDEs/Pb/Cd-contaminated electronic waste site with carboxymethyl chitosan in a sunflower oil-water solvent system and microbial augmentation.

An innovative ex situ soil washing technology was developed to remediate polybrominated diphenyl ethers (PBDEs) and heavy metals in an electronic waste site. Elevated temperature (50 °C) in combination with ultrasonication (40 kHz, 20 min) at 5.0 mL L(-1) sunflower oil and 2.5 g L(-1) carboxymethyl chitosan were found to be effective in extracting mixed pollutants from soil. After two successive washing cycles, the removal efficiency rates for total PBDEs, BDE28, BDE47, BDE209, Pb, and Cd were approximately 94.1, 93.4, 94.3, 99.1, 89.3, and 92.7 %, respectively. Treating the second washed soil with PBDE-degrading bacteria (Rhodococcus sp. strain RHA1) inoculation and nutrient addition for 3 months led to maximum biodegradation rates of 37.3, 52.6, 23.9, and 1.3 % of the remaining total PBDEs, BDE28, BDE47, BDE209, respectively. After the combined treatment, the microbiological functions of washed soil was partially restored, as indicated by a significant increase in the counts, biomass C, N, and functioning diversity of soil microorganisms (p < 0.05), and the residual PBDEs and heavy metals mainly existed as very slow desorbing fractions and residual fractions, as evaluated by Tenax extraction combined with a first-three-compartment model and sequential extraction with metal stability indices (I R and U ts). Additionally, the secondary environmental risk of mixed contaminants in the remediated soil was limited. Therefore, the proposed combined cleanup strategy is an environment-friendly technology that is important for risk assessment and management in mixed-contaminated sites.

Organotropism of persistent organic pollutants and heavy metals in the Greenland shark Somniosus microcephalus in NE Greenland.

The Greenland shark Somniosus microcephalus is an opportunistic feeder, a top predator, and a very long-lived species. The brain, liver, red and white muscle, gonads, fat, skin, pancreas, and spleen of Greenland sharks from NE Greenland fjords were analysed for PCBs, PCDDs/DFs, PBDEs; DDT isomers; HCH isomers; dieldrin; endrin; HCB; Cd, Hg, Pb, and Se. PCBs (2.01-103 ng/g wet wt) and PBDEs (7.9-3050 pg/g wet wt) were detected in most of the samples. PCDDs/DFs showed high values when detected. DDTs, HCB and HCHs were only detected in some tissues. The ΣTEQ was 5.76 pg/g in muscle. Cadmium mainly accumulated in the pancreas and liver (19.6 and 10.7 mg/kg dry wt, respectively); mercury in red muscle (4.10-6.91 mg/kg dry wt); selenium in the pancreas (3.57 mg/kg dry wt) and spleen (1.95 mg/kg dry wt); lead in the skin (0.358 mg/kgd ry wt). The selenium-mercury ratio in the liver was also evaluated.

[Effect of nitrogen and phosphorus concentrations on the adsorption of BDE28 and BDE47 by Prorocentrum donghaiense dominant population].

The adsorption of 2,4, 4'-Tribromodiphenyl ether (BDE28) and 2,2', 4, 4'-Tetrabromodiphenyl ether (BDE47) by Prorocentrum donghaiense dominant population under different nitrogen and phosphorus concentrations was studied. Prorocentrum donghaiense dominant population consisted of Prorocentrum donghaiense and Skeletonema costatum, in which Prorocentrum donghaiense was dominant. Nitrogen was in form of nitrate and the concentrations tested were 0, 128 and 512 micromol x L(-1), and phosphorus was in form of phosphate and the concentrations tested were 0, 8 and 32 micromol x L(-1). The algal density and biochemical composition were determined and the correlation between the adsorption and the biochemical composition was analyzed. The results showed that the BDE28 and BDE47 adsorbed per cell was 2.2 and 2.9 ng x (10(6) cells)(-1) respectively when nitrogen concentration was 0 micromol x L(-1), which was 1.3-1.9 times as high as those under 128 and 512 micromol x L(-1) nitrogen. There was no significant difference in the amount of polybrominated diphenyl ethers (PBDEs) adsorbed per culture and the adsorbed percentage of available PBDEs among different nitrogen concentrations. In nitrogen treatments, the amount of PBDEs adsorbed per culture was 0.160-0.184 ng x mL(-1) and the adsorbed percentage of available PBDEs was 80.2%-92.1%. The BDE28 and BDE47 adsorbed per cell was 7.6 and 8.4 ng x (10(6) cells)(-1) respectively when phosphorus was 0 micromol x L(-1), which was 5.4-6 times as high as those under 8 and 32 micromol x L(-1) phosphorus. There was no significant difference in the amount of PBDEs adsorbed per culture and the adsorbed percentage of available PBDEs among different phosphorus concentrations. In phosphorus treatments, the amount of PBDEs adsorbed per culture was 0.167-0.194 ng x mL(-1) and the adsorbed percentage of available PBDEs was 80.2%-93.2%. Correlation analysis indicated that there was a significant negative correlation between nitrogen concentrations and the amounts of BDE28 and BDE47 adsorbed per cell and a significant positive correlation between phosphorus and the amount of PBDEs adsorbed per culture as well as the adsorbed percentage of available PBDEs. Besides, a significant positive correlation was found between the PBDEs adsorption and the amount of lipids. Nitrogen and phosphorus induced the change of biochemical content which resulted in the change of the adsorption of BDE28 and BDE47.

A polybromodiphenyl ether from an Indonesian marine sponge Lamellodysidea herbacea and its chemical derivatives inhibit protein tyrosine phosphatase 1B, an important target for diabetes treatment.

The ethanol extract of an Indonesian marine sponge Lamellodysidea herbacea inhibited the activity of protein tyrosine phosphatase 1B (PTP1B), an important target enzyme for the treatment of type II diabetes. Bioassay-guided isolation yielded a known polybromodiphenyl ether (1) as a sole bioactive component. The structure of 1 was confirmed by spectroscopic data for 1 and its methyl ether derivative (2). Compound 1 markedly inhibited the PTP1B activity (IC50 = 0.85 µM) and showed a moderate cytotoxicity against two human cancer cell lines, HCT-15 (colon) and Jurkat (T-cell lymphoma) cells. On the other hand, compound 2 maintained potent inhibitory activity against PTP1B (IC50 = 1.7 µM) but did not show apparent cytotoxicity at 18 µM against these cancer cells. Four ester derivatives [acetyl (3), butyryl (4), hexanoyl (5), and benzoyl (6)] were prepared from 1 and their activities evaluated against PTP1B and two cancer cell lines to investigate the structure-activity relationships. Although compounds 3-6 exhibited potent inhibitory effects against PTP1B activity, cytotoxicity against HCT-15 and Jurkat cells was observed as a similar efficacy to that of 1. From these results, compound 2 was found to be the best inhibitor of PTP1B with no apparent cytotoxicity. Therefore, 2 may be a lead compound for making a new type of PTP1B inhibitor. Moreover, compound 2 did not inhibit the cell growth of Huh-7 cells (hepatoma). Hepatocytes are one of the locations of PTP1B, and Huh-7 cells are used to study the mechanism of action of compound 2.

Caged Gammarus fossarum (Crustacea) as a robust tool for the characterization of bioavailable contamination levels in continental waters: towards the determination of threshold values.

We investigated the suitability of an active biomonitoring approach, using the ecologically relevant species Gammarus fossarum, to assess trends of bioavailable contamination in continental waters. Gammarids were translocated into cages at 27 sites, in the Rhône-Alpes region (France) during early autumn 2009. Study sites were chosen to represent different physico-chemical characteristics and various anthropic pressures. Biotic factors such as sex, weight and food availability were controlled in order to provide robust and comparable results. After one week of exposure, concentrations of 11 metals/metalloids (Cd, Pb, Hg, Ni, Zn, Cr, Co, Cu, As, Se and Ag) and 38 hydrophobic organic substances including polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyles (PCBs), pentabromodiphenylethers (PBDEs) and organochlorine pesticides, were measured in gammarids. All metals except Ag, and 33 organic substances among 38 were quantified in G. fossarum, showing that this species is relevant for chemical biomonitoring. The control of biotic factors allowed a robust and direct inter-site comparison of the bioavailable contamination levels. Overall, our results show the interest and robustness of the proposed methodological approach for assessing trends of bioavailable contamination, notably for metals and hydrophobic organic contaminants, in continental waters. Furthermore, we built threshold values of bioavailable contamination in gammarids, above which measured concentrations are expected to reveal a bioavailable contamination at the sampling site. Two ways to define such values were investigated, a statistical approach and a model fit. Threshold values were determined for almost all the substances investigated in this study and similar values were generally derived from the two approaches. Then, levels of contaminants measured in G. fossarum at the 27 study sites were compared to the threshold values obtained using the model fit. These threshold values could serve as a basis for further implementation of quality grids to rank sites according to the extent of the bioavailable contamination, with regard to the applied methodology.

In vitro biotransformation of PBDEs by root crude enzyme extracts: potential role of nitrate reductase (NaR) and glutathione S-transferase (GST) in their debromination.

In order to investigate the enzyme transformation of PBDEs and to track the key enzymes involved in PBDE degradation in plants, in vivo exposure of plants of ryegrass, pumpkin and maize and in vitro exposure of their root crude enzyme extracts to PBDEs were conducted. Degradation of PBDEs in the root crude enzyme solutions fit well with the first order kinetics (R(2)=0.52-0.97, P<0.05), and higher PBDEs degraded faster than the lower ones. PBDEs could be transformed to lower brominated PBDEs and hydroxylated-PBDEs by the root crude enzyme extracts with debromination as the main pathway which contributed over 90% of PBDE depletion. In vitro and in vivo exposure to PBDEs produced similar responses in root enzyme activities of which the nitroreductase (NaR) and glutathione-transferase (GST) activities decreased significantly, while the peroxidase, catalase and cytochrome P-450 activities had no significant changes. Furthermore, higher enzyme concentrations of NaR and GST led to higher PBDE debromination rates, and the time-dependent activities of NaR and GST in the root crude enzyme extracts were similar to the trends of PBDE depletion. All these results suggest that NaR and GST were the key enzymes responsible for PBDE degradation. This conclusion was further confirmed by the in vitro debromination of PBDEs with the commercial pure NaR and GST.

Hydrodebromination of decabromodiphenyl ether (BDE-209) in cooking experiments with salmon fillet.

Polybrominated diphenyl ethers (PBDEs) are environmental contaminants regularly detected in biota and food. Seafood has been identified as the major dietary source for human uptake. Fish is predominantly consumed after cooking, and this process may alter the actual human intake of contaminants. This study thus aimed to investigate the fate of PBDEs in this cooking process. Heating of fish fortified with 2,2',3,3',4,4',5,5',6,6'-decabromodiphenyl ether (BDE-209) at typical cooking conditions (200 °C, in plant oil) resulted in a decrease of its concentration in favor of the formation of lower brominated congeners. After 15 min, ∼25% of BDE-209 was transformed into nona- to octabrominated congeners. The major transformation route was BDE-209 → BDE-206 → BDE-196 and BDE-199. Low amounts of heptabrominated congeners as well as one hexabromodibenzofuran and a heptabromodibenzofuran isomer were also detected. However, penta- and tetrabrominated diphenyl ethers were not observed, and heating of BDE-47 did not produce new transformation products.

Heating of BDE-209 and BDE-47 in plant oil in presence of o,p'-DDT or iron(III) chloride can produce monochloro-polybromo diphenyl ethers.

Heating processes of food can alter the concentrations and composition of organohalogen compounds. In this study the fate of two polybrominated diphenyl ether (PBDE) congeners, decabromodiphenyl ether (BDE-209) and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) was analyzed when heated in plant oil with and without additional compounds. When the PBDEs were heated in pure plant oil, no transformation was observed. Heating of decabromodiphenyl ether (BDE-209) together with ortho,para'-dichlorodiphenyltrichloroethane (o,p'-DDT) or iron(III) chloride in plant oil resulted in the formation of monochloro-nonabromodiphenyl ethers (Br(9)Cl-DEs). Almost 10% of the initial amount of BDE-209 was transferred into Br(9)Cl-DEs. Heating BDE-47 in the presence of iron(III) chloride produced two monochlorinated transformation products which were tentatively identified as 2,2',4-tribromo-4'-chlorodiphenyl ether (4'-Cl-BDE-17) and 2,4,4'-tribromo-2'-chlorodiphenyl ether (2'-Cl-BDE-28). However, the reactivity was smaller and no Br→Cl exchange was observed with o,p'-DDT. The conditions used in our experiments (200 °C; heating 30 min-3 h) indicate that such reactivity may also occur during cooking of fish, meat and other food samples.

Evaluating daily exposure to polychlorinated biphenyls and polybrominated diphenyl ethers in fish oil supplements.

Fish oil supplements have become a popular means of increasing one's dietary intake of essential polyunsaturated fatty acids. However, there is growing concern that the levels and potential health effects of lipophilic organic contaminants such as polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) may diminish some of the health benefits associated with the daily consumption of fish oil supplements. In this study, ten over-the-counter fish oil supplements available in the United States were analysed for PCBs and PBDEs and daily exposures calculated. Based on manufacturers' recommended dosages, daily intakes of PCBs and PBDEs ranged from 5 to 686 ng day(-1) and from 1 to 13 ng day(-1), respectively. Daily consumption of fish oil supplements expose consumers to PCBs and PBDEs. However, in comparison with fish ingestion, fish supplements may decrease daily PCB exposure and provide a safer pathway for individuals seeking to maintain daily recommended levels of polyunsaturated fatty acids.

Gas chromatography-mass spectrometric determination of polybrominated diphenyl ethers in complex fatty matrices from aquaculture activities.

Gas chromatography coupled to mass spectrometry in negative chemical ionization mode (GC-(NCI)MS) has been applied to the quantification and reliable identification of polybrominated diphenyl ethers (PBDEs) in animal and vegetable samples from aquaculture activities. Matrices analyzed included fish fillet, fish feed, fish oil and linseed oil, their fat content ranged from 5% to 100%. Solid-phase extraction (SPE) (using Florisil and silica cartridges) and normal-phase high performance liquid chromatography were tested for an efficient clean-up in order to obtain sample extracts free of interfering compounds. Combining sulphuric acid digestion and SPE with Florisil led to the highest efficiency in the elimination of interferences from the extracts. The sample procedure developed, together with the application of GC-(NCI)MS for measurement, led to the satisfactory determination of PBDEs at microg kg(-1) levels in complex aquaculture matrices with high lipid content. The use of a short and thin film-thickness fused-silica capillary column allowed to determine the problematic BDE 209 with satisfactory results. Three m/z ions were acquired for each analyte, which ensured a reliable identification of compounds detected in samples.

Effects of decontaminated fish oil or a fish and vegetable oil blend on persistent organic pollutant and fatty acid compositions in diet and flesh of Atlantic salmon ( Salmo salar).

The health benefits of seafood are well documented and based on the unique supply of n-3 highly unsaturated fatty acids (HUFA). Aquaculture now contributes about 50 % of food-grade seafood globally and Atlantic salmon (Salmo salar) is a rich source of n-3 HUFA. However, salmon and other oily fish can accumulate lipophilic persistent organic pollutants (POP), including dioxins (PCDD/F), polychlorinated biphenyls (PCB) and polybrominated diphenyl ethers (PBDE), derived largely from feed. In the present study, triplicate groups of salmon, of initial weight 0.78 kg, were fed one of three experimental diets for 11 weeks. The diets were coated with either a northern fish oil (FO) with a high POP content (cNFO), the same oil that had been decontaminated (deNFO) or a blend of southern fish oil, rapeseed and soyabean oils (SFO/RO/SO). Dietary PCDD/F+dioxin-like PCB (DL-PCB) concentrations were 17.36, 0.45 and 0.53 ng toxic equivalents (TEQ)/kg, respectively. After 11 weeks, the flesh concentrations in fish fed the cNFO, deNFO and SFO/RO/SO diets were 6.42, 0.34 and 0.41 ng TEQ/kg, respectively. There were no differences in flesh EPA and DHA between fish fed the cNFO or deNFO diets although EPA and DHA were reduced by 50 and 30 %, respectively, in fish fed the SFO/RO/SO diet. Thus, decontaminated FO can be used to produce salmon high in n-3 HUFA and low in POP. Salmon produced using deNFO would be of high nutritional value and very low in POP and would utilise valuable fish oils that would otherwise be destroyed due to their high pollutant concentrations.

Polybrominated diphenyl ethers: causes for concern and knowledge gaps regarding environmental distribution, fate and toxicity.

This manuscript critically considers several areas of study of the polybrominated diphenyl ether compounds. Specifically, a brief review of PBDE toxicity is followed by an in depth discussion of PBDE occurrence in abiotic and biotic environmental matrices. Temporal and geographic trends are examined in conjunction with risk assessment factors. Rather than summarize or tabulate the growing body of literature on PBDEs in the environment, the overall goal of this review paper is to highlight broad patterns that may contribute to a more holistic understanding of PBDE behavior in the environment, as well as to identify critical areas of research that warrant further attention.