The brominated flame retardant 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE) widely used in manufacturing is inevitably released into the environment, resulting in the exposure of organisms to BTBPE. Therefore, it is particularly important to explore its toxic mechanism. The liver is one of the main accumulating organs of BTBPE, but the mechanism underlying BTBPE hepatotoxicity has not been thoroughly investigated. In our study, BTBPE was administered to Sprague-Dawley (SD) rats and rat hepatocytes (BRL cells) in vivo and in vitro, respectively, and HE staining, AO/EB staining, fluorescent probes, qPCR, immunofluorescence, and dual-luciferase reporter assays were performed. We investigated the mechanism of action of growth arrest-specific 5 (GAS5), miR-743a-5p, and NUAK family kinase 1 (NUAK1) in BTBPE-induced necroptosis from the perspective of competing endogenous RNAs (ceRNAs) using NUAK1 inhibitors, siRNAs, mimics, and overexpression plasmids. Our study showed that exposure to BTBPE caused necroptosis in the liver and BRL cells, accompanied by an oxidation-reduction imbalance and an inflammatory response. It is worth noting that NUAK1 is a newly discovered upstream regulatory target for necroptosis. In addition, miR-743a-5p was shown to inhibit necroptosis by targeting NUAK1 and down-regulating NUAK1. GAS5 upregulates NUAK1 expression by competitively binding to miR-743a-5p, thereby inducing necroptosis. This study demonstrated, for the first time, that the GAS5-miR-743a-5p-NUAK1 axis is involved in the regulation of necroptosis via ceRNAs. Thus, GAS5 and NUAK1 induce necroptosis by competitively binding to miR-743a-5p.

This study explored the effects of 1, 2-bis (2,4, 6-tribromophenoxy) ethane (BTBPE) and bis (2-ethylhexyl) tetrabromophthalate (TBPH) on serum metabolites and lipids in male Sprague-Dawley (SD) rats. Rats were orally gavaged 250 mg/kg bw of BTBPE and 500 mg/kg bw of TBPH for 28 consecutive days. Serum samples were collected for metabolomics and lipidomics analysis. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to explore changes in rat metabolic patterns. Least absolute shrinkage and selection operator (LASSO) regression models were established using serum levels of total thyroxine (TT4), free thyroxine (FT4), and rats\' grouping information as variables to screen for robust differential substances. SuperPred was the database to obtain potential targets. The metabolomics and lipidomics results showed that BTBPE and TBPH had an impact on rat metabolic patterns, affecting pathways such as vitamin B6 synthesis. For BTBPE treatment, pyridoxal and ceramide (Cer) 24:0;4O were selected as differential substances related to thyroid hormones. For TBPH treatment, dehydroascorbic acid, acylcarnitine (CAR) 19:0, and diglyceride (DG) 38:4 were selected as differential substances related to thyroid hormones. Serotonin 2c receptor and cyclooxygenase-2 were chosen as potential targets of BTBPE and TBPH, respectively. In conclusion, this study found that BTBPE and TBPH impacted the metabolism of rats, and this effect may be related to changes in thyroid function.

Novel brominated flame retardants are widely used in electronics, textiles, furniture, and other products; they can enter the human body through ingestion and respiration and cause harm to the human body, and have been proven to have potential biological toxicity and accumulation effects. 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE) is a widely used novel brominated flame retardant; however, there is a lack of research on its mechanism of toxicity, particularly that of intestinal toxicity. Currently, studies on the functionality of iroquois homeobox 3 (IRX3) are extremely limited. In our study, BTBPE was administered to Sprague-Dawley (SD) rats and rat small intestinal crypt epithelial cells (IEC6 cells) in vivo and in vitro, respectively, and hematoxylin and eosin (HE), immunohistochemical, Alcian blue-periodic acid-Schiff (AB-PAS), CCK8, acridine orange/ethidium bromide (AO/EB), fluorescent probes, qPCR, western blotting, and immunofluorescence analyses were performed. To explore the damage mechanism of BTBPE, we used siRNA to silence IRX3 and iNOs-IN-1 (yeast extract-peptone-wheat; YPW) to inhibit nitric oxide synthase 2 (NOS2). The results showed that BTBPE exposure caused inflammation and necroptosis in the jejunum and ileum, as well as destruction of the tight junctions and mucus layer. Moreover, BTBPE activated the IRX3/NOS2 axis both in vivo and in vitro. Silencing IRX3 or inhibiting NOS2 inhibits necroptosis and restores tight junctions in IEC6 cells. In conclusion, our study found that in the jejunum, ileum, and IEC6 cells, BTBPE exposure caused necroptosis and tight junction destruction by activating the IRX3/NOS2 axis. Blocking the IRX3/NOS2 axis can effectively inhibit necroptosis and restore tight junction. In addition, BTBPE exposure caused inflammation and loss of the mucous layer in the jejunum and ileum. Our study is the first to explore the mechanism of intestinal damage caused by BTBPE exposure and to discover new biological functions regulated by the IRX3/NOS2 axis, providing new research directions for necroptosis and tight junctions.

As an alternative to octabromodiphenyl ether (octa-BDE), 1, 2-bis (2,4, 6-tribromophenoxy) ethane (BTBPE) has been widely used in a variety of combustible materials, such as plastics, textiles and furniture. Previous studies have demonstrated the thyroid toxicity of traditional brominated flame retardants for example octa-BDE clearly. Nevertheless, little is known about the thyroid toxicity of alternative novel brominated flame retardants BTBPE. In this study, it was demonstrated that BTBPE in vivo exposure induced FT4 reduction in 2.5, 25 and 250 mg/kg bw treated group and TT4 reduction in 25 mg/kg bw treated group. TG, TPO and NIS are key proteins of thyroid hormone synthesis. The results of Western blot and RT-PCR from thyroid tissue showed decreased protein levels and gene expression levels of TG, TPO and NIS as well as regulatory proteins PAX8 and TTF2. To investigate whether the effect also occurred in humans, anthropogenic Nthy-ori 3-1 cells were selected. Similar results were seen in vitro condition. 2.5 mg/L BTBPE reduced the protein levels of PAX8, TTF1 and TTF2, which in turn inhibited the protein levels of TG and NIS. The results in vitro experiment were consistent with that in vivo, suggesting possible thyrotoxic effects of BTBPE on humans. It was indicated that BTBPE had the potential interference of T4 generation and the study provided more evidence of the effects on endocrine disorders.

Brominated flame retardants such as polybrominated diphenyl ethers (PBDEs) have been used for decades until evidence of negative health effects led to bans in many countries. PBDEs have since been replaced by alternative legacy compounds or newly developed chemicals. In this study, eight alternative brominated flame retardants were analyzed in blubber and liver of harbor seal pups (≤6 months) from the Northwest Atlantic collected during 2001-2010 to elucidate concentrations, patterns, contamination trends, potential maternal transfer, and tissue partitioning. All compounds were detected in liver and blubber tissues with hexabromocyclododecane (HBCD) isomers and 2-ethylhexyl 2,3,4,5-tetrabromobenzoate (TBB) predominating. Overall, α-HBCD was the dominant HBCD isomer in both tissues although the concentrations of γ-HBCD exceeded those of α-HBCD in seven pups, indicating their mothers may have had alternative dietary patterns or recent exposure to the commercial mixture. Although it was detected in less than half of the samples, to our knowledge, this is the first study to report tetrabromobisphenol A (TBBPA) concentrations in multiple tissues of a top marine predator. For the brominated components of Firemaster® flame retardants, TBB concentrations exceeded bis-(2-ethylhexyl)-tetrabromophthalate (TBPH). This pattern may result from recent exposure to commercial mixtures in which TBB exceeds TBPH 4:1 or from differences in perinatal or lactational transfer efficiency of the two compounds. Between the two tissues, lipid-normalized β-HBCD, γ-HBCD, TBB and decabromodiphenyl ethane (DBDPE) concentrations were significantly higher in liver than blubber. This indicates that the bioaccumulation of these chemicals is not simply related to lipid dynamics but may be linked to blood proteins. This study demonstrates that harbor seal pups from this region are contaminated with alternative flame retardants passed to them via placental or lactational transfer. Given the evidence for negative health effects of these chemicals, this contamination adds additional pressure on the first year survival of these young, developing animals.

Global restrictions on use of legacy brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCDD) have generated demand for novel BFRs (NBFRs) as substitutes. Our research group has previously reported decreased concentrations of PBDEs and HBCDD and increased concentrations of NBFRs in UK indoor environments, suggesting that restrictions on PBDEs and HBCDD are exerting an impact. In this study, we analysed UK foodstuffs collected in 2020-21 and compared the BFR concentrations found with those found in similar samples collected in 2015 to investigate whether similar trends in BFR concentrations would be observed. Concentrations of PBDEs and HBCDD isomers detected in our samples had declined by 78-92 % and 59-97 % since the 2015 study, respectively. Moreover, concentrations of NBFRs (dominated by 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE or TBE), and bis(2-ethyl hexyl) tetrabromophthalate (BEH-TEBP or TBPH)) in UK foodstuffs increased significantly (28-1400 %) between 2015 and 2020-21. Combined, these findings suggest that restrictions on use of PBDEs and HBCDD have had a discernible impact on concentrations of these legacy BFRs and their NBFR replacements in UK foodstuffs. Interestingly, given recent reports of a significant increase in concentrations of decabromodiphenyl ethane (DBDPE) in UK house dust between 2014 and 2019, a significant decline (70-84 %) in concentrations of DBDPE was observed in UK foodstuffs.

The novel brominated flame retardants (NBFRs) have become widespread as a consequence of the prohibition on the use of polybrominated diphenyl ethers (PBDEs). However, the transformation mechanism and potential environmental risk are largely unclear. In this study, we have explored the phototransformation behavior of the most abundant NBFRs, 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) in water under ultraviolet (UV) irradiation. Meanwhile, the legacy 2,2\',4,4\',6,6\'-hexabromodiphenyl ether (BDE155) with similar structure was investigated contrastively. Results show that novel BTBPE is more persistent than legacy BDE155, with nearly four times slower photodegradation rate constants (0.0120 min-1and 0.0447 min-1, respectively). 18 products are identified in the phototransformation of BTBPE. Different from the only debrominated products formed in legacy BDE155 transformation, the ether bond cleavage photoproducts (e.g. bromophenols) are also identified in novel BTBPE transformation. Compound-specific stable isotope analysis (CSIA) confirms the phototransformation mechanism is mainly via debromination accompanying with the breaking of ether bond. Computational toxicity assessment implies that transformation products of BTBPE still have the high kidney risks. Especially the bromophenols formed via the ether bond cleavage could significantly increase the health effects on skin irritation. This study emphasizes the importance of understanding the photolytic behavior and potential risks of novel NBFRs and other structurally similar analogues.

Legacy and novel flame retardants (FRs) such as polybrominated diphenyl ethers (PBDEs), novel BFRs, and organophosphate flame retardants (OPFRs) were measured in water and sediment collected from highly industrialized bays of Korea. Predominant compounds in sediment were BDE 209, DBDPE, and BTBPE for BFRs, and TCPP and TBOEP for OPFRs, respectively. Higher alternative FR concentrations were observed compared to those reported for previous studies. The highest BFR concentrations were found in locations close to industrial complexes, while the OPFR concentrations were highest in locations close to domestic regions and a wastewater treatment plant. Different contamination sources were observed for BFRs and OPFRs. The ratio of DBDPE/BDE 209 in sediment ranged from 0.84 to 28, indicating a shift in consumption of BFRs. A significant decline in PBDEs suggests the effectiveness of domestic and global regulations. Despite this, sedimentary PBDE concentrations may pose adverse health risks to benthic organisms and humans.

Polybrominated dibenzo-p-dioxins (PBDDs) and hydroxylated polybrominated diphenyl ethers (OH-PBDEs) can be formed from bromophenols (BPs) by thermal degradation, biosynthesis or phototransformation. However, it is unknown whether PBDDs and OH-PBDEs can be formed during the chemical production processes that utilize BPs as raw materials. 2,4,6-tribromophenol (2,4,6-TBP) is an important raw material for the synthesis of 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE), a novel brominated flame retardant. In this study, PBDDs, polybrominated dibenzofurans (PBDFs) and OH-PBDEs have been identified and quantified in commercially available BTBPE and 2,4,6-TBP. Furthermore, their formation as unintentional by-products during the laboratory synthesis of BTBPE from 2,4,6-TBP and 1,2-dibromoethane in the presence of sodium carbonate has also been investigated. 2,3,7,8-substituted PBDDs and PBDFs (2,3,7,8-PBDD/Fs) were undetectable in commercial samples of BTBPE and present in low levels (nanogram per gram) in 2,4,6-TBP. Two tetrabrominated dibenzo-p-dioxins (TeBDDs), namely 1,3,6,8- and 1,3,7,9-TeBDD, and three hydroxylated pentabrominated diphenyl ethers (OH-pentaBDEs), namely 4\'-OH-BDE121, 2\'-OH-BDE121, and 6\'-OH-BDE100, were identified or tentatively identified, and quantitatively estimated to be at concentrations in the range of undetectable to several thousands of nanograms per gram in commercial BTBPE and 2,4,6-TBP. TeBDDs and OH-pentaBDEs were formed as by-products from 2,4,6-TBP during BTBPE synthesis. Further studies need to be conducted in order to determine whether PBDD/Fs and OH-PBDEs are also formed during the industrial synthesis of other chemical compounds that utilize BPs as raw materials or intermediates.

Brominated flame retardants (BFRs) represent a large group of chemicals used in a variety of household and commercial products to prevent fire propagation. The environmental persistence and toxicity of some of the most widely used BFRs has resulted in a progressive ban worldwide and the development of novel BFRs for which the knowledge on environmental health impacts remains limited. The objectives of this study were to evaluate the effects of two emerging BFRs, 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EH-TBB), in diet exposed juvenile rainbow trout (Oncorhynchus mykiss). Both compounds were detected in fish carcasses at 76% and 2% of the daily dosage of BTBPE and EH-TBB, respectively, indicating accumulation of BTBPE and by contrast extensive depuration/metabolism of EH-TBB. Liver gene transcription analysis using RNA-sequencing indicated that the chronic 28-d dietary exposure of trout to EH-TBB down-regulated one single gene related to endocrine-mediated processes, whereas BTBPE impacted the transcription of 33 genes, including genes involved in the immune response, reproduction, and oxidative stress. Additional analysis using qRT-PCR after 48-h and 28-d of exposure confirmed the impact of BTBPE on immune related genes in the liver (apolipoprotein A-I, lysozyme) and the head-kidney (complement c3-4). However, the activity of lysozymes measured at the protein level did not reflect transcriptomic results. Overall, results suggested an impact on immune-related gene transcription in BTBPE exposed fish, as well as oxidative stress and endocrine disruption potentials.