Potential Link between Equol Pollution and Field-Observed Intersex in Wild So-iuy Mullets (Mugil soiuy).

Gonadal intersex has been observed in wild fishes and is attributed to endocrine-disrupting chemicals but the specific causes remain controversial. Here, a forensic analysis utilizing field and laboratory studies was conducted to explore the causal agent(s). In a 2008-2009 survey of Liaodong Bay, China, 20.7-33.3% incidences of gonadal intersex were observed in male so-iuy mullets (Mugil soiuy), a wild sentinel fish species. Steroidal estrogen (estrone, 17ß-estradiol, estriol, and ethinylestradiol) and phytoestrogen (equol) were detected in seawater where the fishes were collected with median concentrations of 0.42 ng/L (0.02-1.42 ng/L) E2 equivalent (EEQ-E2) and 22.81 ng/L (0.10-155.99 ng/L) equol. A probabilistic model was used to evaluate the ecological risk of these estrogenic chemicals based on their distribution in the field and dose-response relationship from the laboratory surrogate Japanese medaka (Oryzias latipes) fish. The probability of the incidences of gonadal intersex due to equol exposure was estimated to be 13.5 ± 12.1%, which is considerably higher than that for EEQ-E2, (7.2 ± 68.8) × 10-4. The agonistic activity of equol to the estrogen receptor α of so-iuy mullets was 3.5-fold higher than that to the estrogen receptor α of Japanese medaka, indicating that equol shows a stronger potential for inducing intersex in so-iuy mullets than in medaka. These results demonstrate that equol, rather than steroid estrogens, is a more likely causal agent for the field-observed intersex in male wild so-iuy mullets.

Proteome-wide effects of naphthalene-derived secondary organic aerosol in BEAS-2B cells are caused by short-lived unsaturated carbonyls.

Exposure to air pollution causes adverse health outcomes, but the toxicity mechanisms remain unclear. Here, we investigated the dynamic toxicities of naphthalene-derived secondary organic aerosol (NSOA) in a human bronchial epithelial cell line (BEAS-2B) and identified the chemical components responsible for toxicities. The chemical composition of NSOA was found to vary with six simulated atmospheric aging conditions (C1-C6), as characterized by high-resolution mass spectrometry and ion mobility mass spectrometry. Global proteome profiling reveals dynamic evolution in toxicity: Stronger proteome-wide impacts were detected in fresh NSOA, but the effects declined along with atmospheric aging. While Nrf2-regulated proteins (e.g., NQO1) were significantly up-regulated, the majority (78 to 97%) of proteins from inflammation and other pathways were down-regulated by NSOA exposure (e.g., Rho GTPases). This pattern is distinct from the reactive oxygen species (ROS)-mediated toxicity pathway, and an alternative cysteine reaction pathway was revealed by the decreased abundance of proteins (e.g., MT1X) prone to posttranslational thiol modification. This pathway was further validated by observing decreased Nrf2 response in reporter cells, after preincubating NSOA with cysteine. Ethynyl-naphthalene probe was employed to confirm the alkylation of cellular proteome thiols on the proteome-wide level by fresh NSOA via in-gel fluorescence imaging. Nontarget analysis identified several unsaturated carbonyls, including naphthoquinones and hydroxylated naphthoquinones, as the toxic components responsible for cysteine reactivity. Our study provides insights into the dynamic toxicities of NSOA during atmospheric aging and identifies short-lived unsaturated carbonyls as the predominant toxic components at the posttranslational level.

Identification of Chemicals that Cause Oxidative Stress in Oil Sands Process-Affected Water.

Oil sands process-affected water (OSPW) has been reported to cause oxidative stress in organisms, yet the causative agents remain unknown. In this study, a high-throughput in vitro Nrf2 reporter system was used, to determine chemicals in OSPW that cause oxidative stress. Five fractions, with increasing polarity, of the dissolved organic phase of OSPW were generated by use of solid phase extraction cartridges. The greatest response of Nrf2 was elicited by F2 (2.7 ± 0.1-fold), consistent with greater hydroperoxidation of lipids in embryos of Japanese medaka (Oryzias latipes) exposed to F2. Classic naphthenic acids were mainly eluted in F1, and should not be causative chemicals. When F2 was fractionated into 60 subfractions by use of HPLC, significant activation of Nrf2 was observed in three grouped fractions: F2.8 (1.30 ± 0.01-fold), F2.16 (1.34 ± 0.05-fold), and F2.25 (1.28 ± 0.15-fold). 54 compounds were predicted to be potential chemicals causing Nrf2 response, predominated by SO3+ and O3+ species. By use of high-resolution MS2 spectra, these SO3+ and O3+ species were identified as hydroxylated aldehydes. This study demonstrated that polyoxygenated chemicals, rather than classic NAs, were the major chemicals responsible for oxidative stress in the aqueous phase of OSPW.

Hydroxylated 2-Ethylhexyl tetrabromobenzoate isomers in house dust and their agonistic potencies with several nuclear receptors.

In the current study, by combining ultra-high resolution (UHR) MS1 spectra, MS2 spectra, and derivatization, three hydroxylated isomers of 2-ethylhexyl tetrabromobenzoate (OH-TBB) were identified in Firemaster® 550 and BZ-54 technical products. Also, a new LC-UHRMS method, using atmospheric pressure chemical ionization (APCI), was developed for simultaneous analysis of OH-TBB, TBB, hydroxylated bis(2-ethylhexyl)-tetrabromophthalate (OH-TBPH) and TBPH in 23 samples of dust collected from houses in Saskatoon, SK, Canada. OH-TBBs were detected in 91% of samples, with a geometric mean concentration of 0.21 ng/g, which was slightly less than those of OH-TBPH (0.35 ng/g). TBB was detected in 100% of samples of dust with a geometric mean concentration of 992 ng/g. Significant (p < 0.001) log-linear relationships between concentrations of OH-TBBs, TBB, or OH-TBPHs and TBPH in dust support the hypothesis of a common source of these compounds. OH-TBBs were found to be strong agonists of peroxisome proliferator-activated receptor gamma (PPARγ) and weaker agonists of the estrogen receptor (ER), but no agonistic potencies was observed with the androgen receptor (AR). Occurrence of OH-TBBs in technical products and house dust, together with their relatively strong PPARγ potencies, indicated their potential risk to health of humans.

Linking Oxidative Stress and Magnitude of Compensatory Responses with Life-Stage Specific Differences in Sensitivity of White Sturgeon (Acipenser transmontanus) to Copper or Cadmium.

Sensitivity of white sturgeon (Acipenser transmontanus) to copper (Cu) or cadmium (Cd) has been shown to significantly differ as a function of life-stage. This study investigated oxidative stress, metal homeostasis, and associated compensatory responses as potential mechanisms of this sensitivity pattern in three early life-stages. Sturgeon were most sensitive to Cu at 15 days post hatch (dph), which was accompanied by a significant increase in lipid peroxidation (LPO). Genes involved with amelioration of oxidative stress were significantly less inducible at this stage than in older, less sensitive fry. At 48 dph, acute lethality of sturgeon exposed to Cd was greatest and body LPO was significantly induced by 3.5-fold at 5 µg Cd/L. Moreover, there was a small but significant increase in antioxidative responses. At 139 dph, sturgeon were most tolerant to Cu and Cd and accumulation of these metals was least. Also, expression of metallothionein (MT) and apoptotic genes were greatest while expression of metal transporters was reduced and concentration of LPO was not different from controls. Our results suggest that life-stage specific sensitivity of white sturgeon to metals is complex, encompassing differences in the ability to mount compensatory responses important for metal homeostasis and combating oxidative stress and concomitant damages.

Peroxisome Proliferator-Activated Receptor γ is a Sensitive Target for Oil Sands Process-Affected Water: Effects on Adipogenesis and Identification of Ligands.

Identification of toxic components of complex mixtures is a challenge. Here, oil sands process-affected water (OSPW) was used as a case study to identify those toxic components with a known protein target. Organic chemicals in OSPW exhibited dose-dependent activation of peroxisome proliferator-activated receptor γ (PPARγ) at concentrations less than those currently in the environment (0.025× equivalent of full-strength OSPW), by use of a luciferase reporter gene assay. Activation of PPARγ-mediated adipogenesis by OSPW was confirmed in 3T3L1 preadipocytes, as evidenced by accumulation of lipids and up-regulation of AP2, LPL, and PPARγ gene expression after exposure to polar fractions of OSPW. Unexpectedly, the nonpolar fractions of OSPW inhibited differentiation of preadipocytes via activation of the Wnt signaling pathway. Organic chemicals in OSPW that were ligands of PPARγ were identified by use of a pull-down system combined with untargeted chemical analysis (PUCA), with a recombinant PPARγ protein. Thirty ligands of PPARγ were identified by use of the PUCA assay. High resolution MS(1) and MS(2) spectra were combined to predict the formulas or structures of a subset of ligands, and polyoxygenated or heteroatomic chemicals, especially hydroxylated carboxylic/sulfonic acids, were the major ligands of PPARγ.

p,p'-DDE Induces Gonadal Intersex in Japanese Medaka (Oryzias latipes) at Environmentally Relevant Concentrations: Comparison with o,p'-DDT.

Previous studies have reported high body burdens of dichlorodiphenyltrichloroethane (DDT) and its metabolites in wild fishes worldwide. This study evaluated the adverse effects of 1,1-dichloro-2,2-bis (p-chlorophenyl)-ethylene (p,p'-DDE) and o,p'-DDT on gonadal development and reproduction by exposing transgenic Japanese medaka (Oryzias latipes) from hatch for 100 days. While both p,p'-DDE and o,p'-DDT induced intersex in male medaka, the lowest observable effective concentration (LOEC) of o,p'-DDT was 57.7 ng/g ww, about 5-fold lower than that (272 ng/g ww) of p,p'-DDE. Since LOECs of both chemicals were comparable to the body concentrations in wild fish, DDT contamination would likely contribute to the occurrence of intersex observed in wild fish. Exposure to o,p'-DDT resulted in much higher expression of vitellogenin in liver of males than p,p'-DDE, accordant with the higher potency of o,p'-DDT than p,p'-DDE to induce intersex. This phenomenon could be partly explained by the significantly elevated levels of 17ß-estradiol in plasma of males exposed to o,p'-DDT, in addition to its estrogenic activity via the estrogen receptor. Significantly lower fertilization (p = 0.006) and hatchability (p = 0.019) were observed in the 13 intersex males. This study for the first time demonstrated the induction of intersex and reproductive effects of p,p'-DDE and o,p'-DDT at environmentally relevant concentrations.