Human dietary intake and hazard characterization for residues of neonicotinoides and organophosphorus pesticides in Egyptian honey

<p>In two recently published reports, hazards posed by dietary exposure to organophosphate and neonicotinoid plant protection products on the European honey bee (<i>Apis mellifera</i> L.) in Egypt were investigated. Using concentrations reported in those studies, an assessment of hazards posed by these two classes of insecticides to humans due to consumption of Egyptian honey from the Nile Delta during both spring and summer was performed. Twenty-eight compounds including metabolites were assessed for exposure of adult Egyptians based on the best- and worst-case scenarios. Even for the worst-case scenario, exposure to these two classes of pesticides in honey was 15-fold less than hazard index value of 1.0 for adverse effects on humans. Based upon this analysis, people exposed to these insecticides through consumption of honey products would be unlikely to exhibit adverse health outcomes.</p

A Cross-species Quantitative Adverse Outcome Pathway for Activation of the Aryl Hydrocarbon Receptor Leading to Early Life Stage Mortality in Birds and Fishes

Dioxin-like compounds (DLCs) elicit adverse effects through activation of the aryl hydrocarbon receptor (AHR). Prior investigations demonstrated that sensitivity to activation of AHR1 in an in vitro AHR transactivation assay is predictive of early life stage mortality among birds. The present study investigated the link between sensitivity to activation of AHR1s and AHR2s and early life stage mortality among fishes. A significant, linear relationship was demonstrated between sensitivity to activation of AHR2 and early life stage mortality among nine fishes, while no relationship was found for AHR1. The slope and <i>y</i>-intercept for the linear relationship between sensitivity to activation of AHR1 and early life stage mortality in birds was not statistically different from the same relationship for AHR2 in fishes. Data for fishes and birds across DLCs were expanded into four significant, linear regression models describing the relationship between sensitivity to activation of AHR and the dose to cause early life stage mortality of 0%, 10%, 50%, or 100%. These four relationships were combined to form a quantitative adverse outcome pathway which can predict dose–response curves of early life stage mortality for DLCs to any bird or fish from species- and chemical-specific responses in an in vitro AHR transactivation assay

A Cross-species Quantitative Adverse Outcome Pathway for Activation of the Aryl Hydrocarbon Receptor Leading to Early Life Stage Mortality in Birds and Fishes

Dioxin-like compounds (DLCs) elicit adverse effects through activation of the aryl hydrocarbon receptor (AHR). Prior investigations demonstrated that sensitivity to activation of AHR1 in an in vitro AHR transactivation assay is predictive of early life stage mortality among birds. The present study investigated the link between sensitivity to activation of AHR1s and AHR2s and early life stage mortality among fishes. A significant, linear relationship was demonstrated between sensitivity to activation of AHR2 and early life stage mortality among nine fishes, while no relationship was found for AHR1. The slope and <i>y</i>-intercept for the linear relationship between sensitivity to activation of AHR1 and early life stage mortality in birds was not statistically different from the same relationship for AHR2 in fishes. Data for fishes and birds across DLCs were expanded into four significant, linear regression models describing the relationship between sensitivity to activation of AHR and the dose to cause early life stage mortality of 0%, 10%, 50%, or 100%. These four relationships were combined to form a quantitative adverse outcome pathway which can predict dose–response curves of early life stage mortality for DLCs to any bird or fish from species- and chemical-specific responses in an in vitro AHR transactivation assay

Effects of Exposure to 17α-Ethynylestradiol during Sexual Differentiation on the Transcriptome of the African Clawed Frog (<i>Xenopus laevis</i>)

Exposure to estrogens during the period of sexual differentiation is known to adversely affect the development of testes in African clawed frogs (<i>Xenopus</i> laevis), but little is known about molecular changes that coincide with the development of altered phenotypes. Therefore, the transcriptome-level effects of exposure to 17α-ethynylestradiol (EE2) during sexual differentiation of <i>X. laevis</i> were evaluated by use of <i>Illumina</i> sequencing coupled with RNA-Seq expression analysis. Overall, a number of processes were affected by 17α-ethynylestradiol, including steroid biosynthesis, thyroid hormone signaling and metabolism, testicular development, and spermatogenesis. Some of the altered pathways, such as thyroid hormone signaling and testicular development, could be linked with biological effects on metamorphosis and gonadal phenotypes, respectively, that were observed in frogs that were exposed to 17α-ethynylestradiol throughout metamorphosis and the early postmetamorphic period. Thus, early changes at the transcriptome-level were predictive of pathologies that did not manifest until later in development. To validate the quantitative capacity of RNA-Seq, a subset of transcripts identified to have altered abundances in individuals exposed to 17α-ethynylestradiol was also evaluated by use of quantitative polymerase chain reaction (qPCR). While small sample sizes (<i>n</i> = 3) limited the ability to draw conclusions pertaining to differences in qPCR-derived abundances of transcripts between control and exposed tadpoles, there was a significant relationship (<i>r</i>² = 0.78) between fold-changes for RNA-Seq and qPCR

A Reagent-Free Screening Assay for Evaluation of the Effects of Chemicals on the Proliferation and Morphology of HeLa-GFP Cells

A reagent-free screening assay using HeLa cells with green fluorescent protein expressed in the cytoplasm was developed to describe adherence and proliferation of cells after seeding and to evaluate the dose- and time-dependent effects of three classes of chemicals, including metals (CdCl₂, MeHg, HgCl₂, and NiCl₂), flame retardants [tris­(1,3-dichloro-2-propyl) phosphate, heptadecafluoro-1-octanesulfonic acid, tetrabromobisphenol A, and tris­(2-chloroethyl) phosphate], and phenolic fungicides (2,4,6-trichlorophenol, 4-<i>tert</i>-butylphenol, 3-bromophenol, and 2,4-dibromophenol), on the proliferation and morphology of cells. A seeding density of 5000 cells/well was determined to be optimal, and the most suitable duration of xenobiotic exposure was 120 h beginning 58 h postseeding. Dose- and time-dependent alterations in total fluorescence in the fluorescent field and number of cells, area per cell, and cellular roundness in the bright field were identified after exposure to each of the target chemicals. Results were comparable to those of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay or other end points published in the literature for the same chemicals

Responses of the Proteome and Metabolome in Livers of Zebrafish Exposed Chronically to Environmentally Relevant Concentrations of Microcystin-LR

In this study, for the first time, changes in expressions of proteins and profiles of metabolites in liver of the small, freshwater fish Danio rerio (zebrafish) were investigated after long-term exposure to environmentally relevant concentrations of microcystin-LR (MC-LR). Male zebrafish were exposed via water to 1 or 10 μg MC-LR/L for 90 days, and iTRAQ-based proteomics and ¹H NMR-based metabolomics were employed. Histopathological observations showed that MC-LR caused damage to liver, and the effects were more pronounced in fish exposed to 10 μg MC-LR/L. Metabolomic analysis also showed alterations of hepatic function, which included changes in a number of metabolic pathways, including small molecules involved in energy, glucose, lipids, and amino acids metabolism. Concentrations of lactate were significantly greater in individuals exposed to MC-LR than in unexposed controls. This indicated a shift toward anaerobic metabolism, which was confirmed by impaired respiration in mitochondria. Proteomics revealed that MC-LR significantly influenced multiple proteins, including those involved in folding of proteins and metabolism. Endoplasmic reticulum stress contributed to disturbance of metabolism of lipids in liver of zebrafish exposed to MC-LR. Identification of proteins and metabolites in liver of zebrafish responsive to MC-LR provides insights into mechanisms of chronic toxicity of MCs

Identification of Thyroid Hormone Disruptors among HO-PBDEs: <i>In Vitro</i> Investigations and Coregulator Involved Simulations

Some hydroxylated polybrominated diphenyl ethers (HO-PBDEs), that have been widely detected in the environment and tissues of humans and wildlife, bind to thyroid hormone (TH) receptor (TR) and can disrupt functioning of systems modulated by the TR. However, mechanisms of TH disrupting effects are still equivocal. Here, disruption of functions of TH modulated pathways by HO-PBDEs was evaluated by assays of competitive binding, coactivator recruitment, and proliferation of GH3 cells. <i>In silico</i> simulations considering effects of coregulators were carried out to investigate molecular mechanisms and to predict potencies for disrupting functions of the TH. Some HO-PBDEs were able to bind to TR with moderate affinities but were not agonists. In GH3 proliferation assays, 13 out of 16 HO-PBDEs were antagonists for the TH. <i>In silico</i> simulations of molecular dynamics revealed that coregulators were essential for identification of TH disruptors. Among HO-PBDEs, binding of passive antagonists induced repositioning of H12, blocking AF-2 (transactivation function 2) and preventing recruitment of the coactivator. Binding of active antagonists exposed the coregulator binding site, which tended to bind to the corepressor rather than the coactivator. By considering both passive and active antagonisms, anti-TH potencies of HO-PBDEs could be predicted from free energy of binding

Mechanisms of Toxicity of Hydroxylated Polybrominated Diphenyl Ethers (HO-PBDEs) Determined by Toxicogenomic Analysis with a Live Cell Array Coupled with Mutagenesis in Escherichia coli

Results of previous studies have indicated that 6-HO-BDE-47, the addition of the hydroxyl (HO) group to the backbone of BDE-47, significantly increased the toxicity of the chemical compared to its postulated precursor analogues, BDE-47 and 6-MeO-BDE-47. However, whether such a result is conserved across polybrominated diphenyl ether (PBDE) congeners was unknown. Here, cytotoxicity of 32 PBDE analogues (17 HO-PBDEs and 15 MeO-PBDEs) was further tested and the underlying molecular mechanism was investigated. A total of 14 of the 17 HO-PBDEs inhibited growth of Escherichia coli during 4 or 24 h durations of exposure, but none of the MeO-PBDEs was cytotoxic at the concentrations tested. 6-HO-BDE-47 and 2-HO-BDE-28 were most potent with 4 h median effect concentrations (EC₅₀) of 12.13 and 6.25 mg/L, respectively, which trended to be lesser with a longer exposure time (24 h). Expression of 30 modulated and validated genes by 6-HO-BDE-47 in a previous study was also observed after exposure to other HO-PBDE analogues. For instance, <i>uhpT</i> was upregulated by 13 HO-PBDEs, and three rRNA operons (<i>rrnA</i>, <i>rrnB</i>, and <i>rrnC</i>) were downregulated by 8 HO-PBDEs. These unanimous responses suggested a potential common molecular signaling modulated by HO-PBDEs. To explore new information on mechanisms of action, this work was extended by testing the increased susceptibility of 182 mutations of transcriptional factors (TFs) and 22 mutations as genes modulated by 6-HO-BDE-47 after exposure to 6-HO-BDE-47 at the 4 h IC₅₀ concentration. Although a unanimous upregulation of <i>uhpT</i> was observed after exposure to HO-PBDEs, no significant shift in sensitivity was observed in <i>uhpT</i>-defective mutants. The 54 genes, selected by cut-offs of 0.35 and 0.65, were determined to be responsible for “organic acid/oxoacid/carboxylic acid metabolic process” pathways, which supported a previous finding

Isomer-Specific Accumulation of Perfluorooctanesulfonate from (<i>N</i>‑Ethyl perfluorooctanesulfonamido)ethanol-based Phosphate Diester in Japanese Medaka (Oryzias latipes)

While (<i>N</i>-ethyl perfluorooctanesulfonamido)­ethanol (FOSE) -based phosphate diester (diSPAP) has been proposed as a candidate precursor of perfluorooctanesulfonate (PFOS), its potential biotransformation to PFOS has not been verified. Metabolism of diSPAP was investigated in Japanese medaka (Oryzias latipes) after exposure in water for 10 days, followed by 10 days of depuration. Branched isomers of diSPAP (B-diSPAP) were preferentially enriched in medaka exposed to diSPAP, with the proportion of branched isomers (BF) ranging from 0.56 to 0.80, which was significantly greater than that in the water to which the medaka were exposed (0.36) (<i>p</i> < 0.001). This enrichment was due primarily to preferential uptake of B-diSPAP. PFOS together with perfluorooctanesulfonamide (PFOSA), <i>N</i>-ethyl perfluorooctanesulfonamide (NEtFOSA), 2-(perfluorooctanesulfonamido)­acetic acid (FOSAA), NEtFOSAA, FOSE, and NEtFOSE were detected in medaka exposed to diSPAP, which indicated the potential for biotransformation of diSPAP to PFOS via multiple intermediates. Due to preferential metabolism of branched isomers, FOSAA and PFOSA exhibited greater BF values (>0.5) than those of NEtFOSA, NEtFOSAA, and NEtFOSE (<0.2). Such preferential metabolism of branched isomers along the primary pathway of metabolism and preferential accumulation of B-diSPAP led to enrichment of branched PFOS (B-PFOS) in medaka. Enrichment of B-PFOS was greater for 3-, 4-, and 5-perfluoromethyl PFOS (P₃MPFOS, P₄MPFOS, and P₅MPFOS), for which values of BF were 0.58 ± 0.07, 0.62 ± 0.06, and 0.61 ± 0.05 (day 6), respectively; these values are 5.8-, 7.8-, and 6.4-fold greater than those of technical PFOS. This work provides evidence on the isomer-specific accumulation of PFOS from diSPAP and will be helpful to track indirect sources of PFOS in the future

Occurrences and Fates of Hydroxylated Polybrominated Diphenyl Ethers in Marine Sediments in Relation to Trophodynamics

While occurrences and origins of hydroxylated (OH-) polybrominated diphenyl ethers (PBDEs) in organisms have been reported, the fates of these compounds in abiotic matrixes and related trophodynamics are unclear. The present study measured concentrations of nine OH-PBDEs, twelve methoxylated (MeO-) PBDEs, and eleven PBDEs in marine sediments and explored the trophodynamics of OH-PBDEs in five invertebrates, eight fish, and two species of birds from Liaodong Bay, north China. While concentrations of PBDEs were less than the limit of quantification in sediments, concentrations of ΣOH-PBDEs and ΣMeO-PBDEs were 3.2–116 pg/g dry weight (dw) and 3.8–56 pg/g dw, respectively. When the detected compounds were incubated in native marine sediments the interconversion between 6-OH-BDE47 and 6-MeO-BDE47 was observed. This result is consistent with the similar spatial distributions and significant correlation between the concentrations of these naturally occurring compounds. 6-OH-BDE47 and 2′-OH-BDE68 were detected as the two major congeners in organisms collected from Liaodong Bay, and concentrations were 0.24 ± 0.005 ng/g lw (lipid weight) and 0.088 ± 0.006 ng/g lw, respectively. Biota-sediment accumulation factors (BSAFs) for invertebrates of 6-OH-BDE47 and 2′-OH-BDE68 were 0.017–0.96 and 0.19–1.5 (except for short-necked clam: 6.3), respectively. Lipid-normalized concentrations of 6-OH-BDE47 and 2′-OH-BDE68 decreased significantly with trophic level with TMFs of 0.21 and 0.15, respectively. The fates of OH-PBDEs in sediment together with their trophodynamics in marine food webs suggested that OH-PBDEs are partitioned into sediment and undergo biodilution in the marine food web