Screening Novel Per- and Polyfluoroalkyl Substances in Human Blood Based on Nontarget Analysis and Underestimated Potential Health Risks.

Nontarget analysis has gained prominence in screening novel perfluoroalkyl and polyfluoroalkyl substances (PFASs) in the environment, yet remaining limited in human biological matrices. In this study, 155 whole blood samples were collected from the general population in Shijiazhuang City, China. By nontarget analysis, 31 legacy and novel PFASs were assigned with the confidence level of 3 or above. For the first time, 11 PFASs were identified in human blood, including C1 and C3 perfluoroalkyl sulfonic acids (PFSAs), C4 ether PFSA, C8 ether perfluoroalkyl carboxylic acid (ether PFCA), C4-5 unsaturated perfluoroalkyl alcohols, C9-10 carboxylic acid-perfluoroalkyl sulfonamides (CA-PFSMs), and C1 perfluoroalkyl sulfonamide. It is surprising that the targeted PFASs were the highest in the suburban population which was impacted by industrial emission, while the novel PFASs identified by nontarget analysis, such as C1 PFSA and C9-11 CA-PFSMs, were the highest in the rural population who often drank contaminated groundwater. Combining the toxicity prediction results of the bioaccumulation potential, lethality to rats, and binding affinity to target proteins, C3 PFSA, C4 and C7 ether PFSAs, and C9-11 CA-PFSMs exhibit great health risks. These findings emphasize the necessity of broadening nontarget analysis in assessing the PFAS exposure risks, particularly in rural populations.

Temozolomide-based sonodynamic therapy induces immunogenic cell death in glioma.

BACKGROUND: In our previous study, we found for the first time that temozolomide (TMZ), the first-line chemotherapeutic agent for glioblastoma (GBM), can generate a large amount of reactive oxygen species (ROS) under ultrasound irradiation. Sonodynamic therapy (SDT) using TMZ as the sonosensitizer produced more potent antitumor effects than TMZ alone. Here, we further evaluate the effects of TMZ-based SDT on subcellular structures and investigate the immunogenic cell death (ICD)-inducing capability of TMZ-based SDT. METHODS: The sonotoxic effects of TMZ were explored in LN229 and GL261 glioma cells. The morphology of endoplasmic reticulum and mitochondria was observed by transmission electron microscopy. The nuclear DNA damage was represented by γ-H2AX staining. Bone marrow-derived dendritic cells (BMDCs) were employed to assess ICD-inducing capability of TMZ-based SDT. A cyclic arginine-glycine-aspartic (c(RGDyC))-modified nanoliposome drug delivery platform was used to improve the tumor targeting of SDT. RESULTS: TMZ-based SDT had a greater inhibitory effect on glioma cells than TMZ alone. Transmission electron microscopy revealed that TMZ-based SDT caused endoplasmic reticulum dilation and mitochondrial swelling. In addition, endoplasmic reticulum stress response (ERSR), nuclear DNA damage and mitochondrial permeability transition pore (mPTP) opening were promoted in TMZ-based SDT group. Most importantly, we found that TMZ-based SDT could promote the "danger signals" produced by glioma cells and induce the maturation and activation of BMDCs, which was associated with the mitochondrial DNA released into the cytoplasm in glioma cells. In vivo experiments showed that TMZ-based SDT could remodel glioma immune microenvironment and provoke durable and powerful anti-tumor immune responses. What's more, the engineered nanoliposome vector of TMZ conferred SDT tumor targeting, providing an option for safer clinical application of TMZ in combination with SDT in the future. CONCLUSIONS: TMZ-based SDT was capable of triggering ICD in glioma. The discovery of TMZ as a sonosensitizer have shown great promise in the treatment of GBM.

2-Ethylhexyl Diphenyl Phosphate Causes Obesity in Zebrafish by Stimulating Overeating via Inhibition of Dopamine Receptor D2.

Obesity is a popular public health problem worldwide and is mainly caused by overeating, but little is known about the impacts of synthetic chemicals on obesity. Herein, we evaluated the obesogenic effect caused by 2-ethylhexyl diphenyl phosphate (EHDPHP) on zebrafish. Adult zebrafish were exposed to 5, 35, and 245 µg/L of EHDPHP for 21 days. Results showed that EHDPHP exposure significantly promoted the feeding behavior of zebrafish, as evidenced by shorter reaction time, increased average food intake, feeding rate, and intake frequency (p < 0.05). Transcriptomic, real-time quantitative PCR, and neurotransmitter analyses revealed that the dopamine (DA) receptor D2 (DRD2) was inhibited, which interfered with the DA neural reward regulation system, thus stimulating food addiction to zebrafish. This was further verified by the restored DRD2 after 7 days of Halo (a DRD2 agonist) treatment. A strong interaction between EHDPHP and DRD2 was identified via molecular docking. As a consequence of the abnormal feeding behavior, the exposed fish exhibited significant obesity evidenced by increased body weight, body mass index, plasma total cholesterol, triglyceride, and body fat content. Additionally, the pathways linked to Parkinson's disease, alcoholism, and cocaine addiction were also disrupted, implying that EHDPHP might cause other neurological disorders via the disrupted DA system.

First insight into the sex-dependent accumulation, tissue distribution and potential toxicities of 2-ethylhexyl diphenyl phosphate and its metabolites in adult zebrafish.

The 2-ethylhexyl diphenyl phosphate (EHDPHP), a primary organophosphorus flame retardant used in various industrial products, is prone to biotransformation. However, there is a knowledge gap on the sex- and tissue-specific accumulation and potential toxicities of EHDPHP (M1) and its metabolites (M2-M16). In this study, adult zebrafish (Danio rerio) were exposed to EHDPHP (0, 5, 35 and 245 µg/L) for 21-day, which was followed by 7-day depuration. The bioconcentration factor (BCF) of EHDPHP in female zebrafish was 26.2 ± 7.7% lower than in males due to the lower uptake rate (ku) while higher depuration rate (kd) in the females. The regular ovulation and higher metabolic efficiency promoted elimination from female zebrafish, thus leading to much less (28-44%) accumulation of ∑(M1-M16) in female zebrafish. They exhibited the highest accumulation in the liver and intestine in both sexes, which might be regulated by tissue-specific transporters and histones evidenced by molecular docking results. Intestine microbiota analysis further revealed that female zebrafish were more susceptible to EHDPHP exposure, with more significant changes in phenotype number and KEGG pathways in female than male fish. Disease prediction results suggested that EHDPHP exposure might cause cancers, cardiovascular diseases as well as endocrine disorders in both sexes. These results provide a comprehensive understanding of the sex-dependent accumulation and toxicity of EHDPHP and its metabolites.

Microbial biotransformation mechanisms of PFPiAs in soil unveiled by metagenomic analysis.

As alternatives of long-chain PFASs (Poly- and perfluoroalkyl substances), perfluoroalkyl phosphinic acids (PFPiAs) are increasingly observed in the environment, but their environmental behaviors have not been well understood. Here, the microbial biotransformation of C6/C6 and C8/C8 PFPiA in two soils (Soil N and Y) was investigated. After 252 d and 330 d of incubation with PFPiAs in Soil N and Y respectively, the levels of PFPiAs decreased distinctly, accompanied by the increasing perfluorohexaphosphonic acid (PFHxPA) or perfluorooctanophosphonic acid (PFOPA) formation, magnifying PFPiAs were susceptible to C-P cleavage, which was also confirmed by the density functional theory calculations. The half-lives of the PFPiAs were longer than one year, while generally shorter in Soil N than in Soil Y and that of C6/C6 was shorter than C8/C8 PFPiA (392 d and 746 d in Soil N, and 603 and 1155 d in Soil Y, respectively). Metagenomic sequencing analysis revealed that Proteobacteria as the primary host of the potential functional genes related to CP bond cleavage might be the crucial phyla contributing to the biotransformation of PFPiAs. Meanwhile, the more intensive interactions between the microbes in Soil N consistently contribute to its greater capacity for transforming PFPiAs.

First Insight into the Formation of In Vivo Transformation Products of 2-Ethylhexyl diphenyl phosphate in Zebrafish and Prediction of Their Potential Toxicities.

As a frequently detected organophosphorus flame retardant in the environment, 2-ethylhexyl diphenyl phosphate (EHDPHP) is vulnerable to biotransformation, while the transformation mechanisms and potential toxicities of its transformation products remain unclear. In the present study, in vivo transformation products of EHDPHP in exposed zebrafish for 21d were analyzed by suspect screening and identified by mass spectrometry. Fifteen metabolites were identified, including 10 phase I and 5 phase II products with monohydroxylated products being primary, among which 5-OH-EHDPHP was the most predominant. Two sulfation products and one terminal desaturation metabolite of EHDPHP were reported for the first time. A density functional calculation coupled with molecular docking disclosed that the specific conformation of EHDPHP docked in the protein pockets favored the primary formation of 5-OH-EHDPHP, which was fortified to be a more suitable biomarker of EHDPHP exposure. The in vitro tests suggested that EHDPHP transformation took place not only in liver but also in intestine, where gut microbes played an important role. Due to lack of standards, in silico toxicity prediction combined with molecular docking indicated that several metabolites potentially cause higher toxicities than EHDPHP. The results provide deep insight into the potential health risks due to specific in vivo transformation of EHDPHP.

Insights into the lower trophic transfer of silver ions than silver containing nanoparticles along an aquatic food chain.

Silver nanoparticles (AgNPs) released into the environment are subject to environmental transformation processes before accumulating in aquatic organisms and transferring along the food chain. Lack of understanding on how environmental transformation affects trophic transfer of AgNPs hinders accurate prediction of the environmental risks of these widely present nanomaterials. Here we discover that pristine AgNPs as well as their sulfidation products (Ag2S-NPs) and dissolution products (Ag+) tend to be accumulated in Daphnia magna and subsequently transferred to zebrafish. In D. magna, Ag+ exhibits the highest bioaccumulation potential whereas Ag2S-NPs show the lowest bioaccumulation. Surprisingly, the biomagnification factor of Ag+ along the D. magna-zebrafish food chain appears to be significantly lower relative to AgNPs and Ag2S-NPs, likely due to the limited release of Ag from D. magna to zebrafish during digestion. Moreover, AgNPs and their transformation products mainly accumulate in the internal organs, particularly intestine, of zebrafish. Adsorption of AgNPs on the surface of the intestinal cell membrane mitigates depuration of AgNPs and, at least in part, leads to the larger biomagnification factor of AgNPs, relative to their transformation products. This research highlights the necessity of considering environmental transformation processes of nanomaterials in assessing their bioavailability and risk.

Insights into the sex-dependent reproductive toxicity of 2-ethylhexyl diphenyl phosphate on zebrafish (Danio rerio).

As a frequently detected organophosphate ester in various environmental media, the toxic effects of 2-ethylhexyl diphenyl phosphate (EHDPHP) on aquatic organisms of different sexes remain unclear. In this study, adult zebrafish were exposed to 2.5, 50, 250 µg/L of EHDPHP for 21 days to investigate its sex-dependent reproductive toxicity and related mechanisms. EHDPHP exposure significantly inhibited the reproduction of zebrafish, evidenced by the reduced spawning of females, depressed growth and development of their offspring. EHDPHP induced greater impacts on the changes of sex hormones and vitellogenin (VTG) in the males than females. For females, the synthesis of testosterone (T) was inhibited because of the down-regulated gnrhr2, gnrhr3, gnrhr4, gnrh3, gnrh2 and er2ß in the brain, while 17ß-estradiol (E2) increased in 250 µg/L due to up-regulated cyp19a. For males, the promotion of T was directly related to the up-regulation of fshr, 3ßhsd, star, cyp11 and cyp17 in the gonad, and eventually led to the increase of E2. The decrease of plasma 11-KT in both sexes could be mainly attributed to the down-regulation of cyp11b and hsd11b. The plasma VTG decreased in females but increased in males, which was in accordance with the down and up regulation of erα and er2ß in the females and males, respectively. All these indicated EHDPHP displayed reproductive toxicity on zebrafish in a sex dependent manner. Molecular docking analysis indicated stronger interaction of EHDPHP with the antagonisms of estrogen receptor (ER) and androgen receptor (AR), as well as the agonism of CYP19A1, which further revealed the sex-dependent reproductive toxicity mechanism of EHDPHP. This study highlights the importance of distinguishing males and females in toxicity evaluation of endocrine disruption chemicals.

Distribution and sources of ordinary monomeric and emerging oligomeric organophosphorus flame retardants in Haihe Basin, China.

With the banning of poly-brominated diphenyl ethers, organophosphorus flame retardants (OPFRs) have gained great development space as their alternatives. In this study, the concentration and partition of nine ordinary monomeric OPFRs and two emerging oligomeric OPFRs (Resorcinol bis(diphenyl phosphate) (RDP) and bisphenol A bis(diphenyl phosphate) (BDP) in the Haihe Basin of China were studied, and their possible sources were analyzed. The Æ©OPFRs in the surface water, sediments and suspended particulate matters (SPMs) in Haihe Basin were 80.1-777 ng/L, 18.9-86.5 ng/g dw and 6.40-234 ng/g dw, respectively. The detection frequencies of both oligomeric OPFRs exceeded 85%, suggesting their environmental contamination was widespread. The mean concentrations of RDP and BDP were 2.09 and 6.03 ng/L in the surface water, 0.91 and 5.09 ng/g dw in the sediments, 1.08 and 2.61 ng/g dw in SPMs, respectively. With Cl-OPFRs being predominant in all the three media, the proportions of alkyl-OPFRs, aryl-OPFRs, and oligomeric OPFRs in the sediments and SPMs were remarkably higher than those in surface water, suggesting that non-halogenated OPFRs preferred to distribute in solid phases. The water-sediment/SPM partition coefficients, logKoc, of the OPFRs were calculated based on the paired samples. The logKoc values of RDP and BDP were reported for the first time, and their logKoc water-sediment were 4.36 ± 0.47 and 4.76 ± 0.25 and their logKoc water-SPM were 3.77 ± 0.45 and 4.00 ± 0.47, respectively. The obtained logKoc values were correlated with their octanol-water partition coefficient (logKow) significantly. It suggested that their partition to sediment or SPM was dominated by hydrophobic interaction. Principal component analysis indicated that the emissions from manufacturing plants and airport operation might be two important sources of OPFRs in the Haihe Basin of China.

Uptake Kinetics, Accumulation, and Long-Distance Transport of Organophosphate Esters in Plants: Impacts of Chemical and Plant Properties.

The uptake, accumulation, and long-distance transport of organophosphate esters (OPEs) in four kinds of plants were investigated by hydroponic experiments. The uptake kinetics ( k1,root) of OPEs in plant roots were determined by the binding of OPEs with the proteins in plant roots and apoplastic sap for the hydrophobic compounds, which correlated well with the transpiration capacity of the plants for the hydrophilic compounds. However, the accumulation capacity of OPEs in plant root was controlled by the partition of OPEs to plant lipids. As a consequence, OPEs were taken up the fastest in wheat root as a result of its highest protein content but least accumulated as a result of its lowest lipid content. The translocation factor of the OPEs decreased quickly with the hydrophobicity (log Kow) increasing, suggesting that the hydrophobic OPEs were hard to translocate from roots to shoots. The hydrophilic OPEs, such as tris(2-chloroisopropyl) phosphate and tris(2-butoxyethyl) phosphate, were ambimobile in the plant xylem and phloem, suggesting that they could move to the edible parts of plants and enhanced risk to human health.

Health risk assessment of heavy metals in freshwater fish in the central and eastern North China.

The distribution and potential health risks of eight heavy metals (Copper (Cu), Chromium (Cr), Zinc (Zn), Lead (Pb), Arsenic (As), Cadmium (Cd), Manganese (Mn), Nickel (Ni)) in 16 freshwater systems from central and eastern North China, were investigated. The fish were divided as wild fish, which grew naturally without artificially feeding, and farmed fish. The total concentrations of the eight heavy metals ranged from 82.9 to 226 µg/L in the surface water samples and 3.32-27.6 mg/kg dw in the fish samples. There was no significant difference in the heavy metal concentrations between natural and farmed water systems. The concentrations of toxic metals, including Pb, As, Cd, Cr, are similar in all kinds of fish. However, the essential metals (Zn, Cu, Mn, Ni) in crucian carp (15.9 mg/kg) was much higher than other kinds of fish. Comparing the wild and farmed fish, the average concentrations of each heavy metal in wild crucian carp, bighead carp, grass carp were higher than those in farmed fish. The average log BCFs (bioconcentration factor) of Zn, Cr and Cu were the highest (2.14, 2.04, 2.00 L/kg) while that of Cd (0.65 L/kg) was the lowest. The non-carcinogenic and carcinogenic health risks to adults and children resulting from consuming the fish were assessed based on the target hazard quotients (THQ). The results indicated that the non-carcinogenic health risk to humans by consuming fish products, no matter wild or farmed fish, was relatively low. The carcinogenic risk of inorganic As was 5.11 × 10-6-1.95 × 10-4 for children and 2.71 × 10-6-1.04 × 10-4 for adult, which are within the acceptable range. The results indicated that the concentrations of heavy metals in the freshwater fish in central and eastern North China were relatively low, and did not cause considerable human health risks.