ABSTRACT
Humans are exposed to a cocktail of food-related and environmental contaminants, potentially contributing to the etiology of chronic diseases. Better characterizing the "exposome" is a challenging task and requires broad human biomonitoring (HBM). Veterinary drugs (VDs)/antibiotics, widely used and regulated in food and animal production, however, are typically not yet included in exposomics workflows. Therefore, in this work, a previously established multianalyte liquid chromatography-tandem mass spectrometry (LC-MS/MS) method covering >80 diverse xenobiotics was expanded by >40 VDs/antibiotics and pesticides. It was investigated if the generic workflow allowed for the successful integration of a high number of new analytes in a proof-of-principle study. The expanded method was successfully in-house validated and specificity, matrix effects, linearity, intra- and inter-day precision, accuracy, limits of quantification, and detection were evaluated. The optimized method demonstrated satisfactory recovery (81-120%) for most of the added analytes with acceptable RSDs (<20%) at three spiking levels. The majority of VDs/antibiotics and pesticides (69%) showed matrix effects within a range of 50-140%. Moreover, sensitivity was excellent with median LODs and LOQs of 0.10 ng/mL and 0.31 ng/mL, respectively. In total, the expanded method can be used to detect and quantify more than 120 highly diverse analytes in a single analytical run. To the best of the authors' knowledge, this work represents the first targeted biomonitoring method integrating VDs with various other classes of pollutants including plasticizers, PFAS, bisphenols, mycotoxins, and personal care products. It demonstrates the potential to expand targeted multianalyte methods towards additional groups of potentially toxic chemicals.
Subject(s)
Biological Monitoring , Pesticides , Veterinary Drugs , Animals , Humans , Biological Monitoring/methods , Exposome , Limit of Detection , Liquid Chromatography-Mass Spectrometry , Pesticides/analysis , Reproducibility of Results , Tandem Mass Spectrometry/methods , Veterinary Drugs/analysisABSTRACT
Metastasis is the major cause of high mortality in lung cancer. Exploring the underlying mechanisms of metastasis thus holds promise for identifying new therapeutic strategies that may enhance survival. Methods: We applied quantitative mass spectrometry to compare protein expression profiles between primary and metastatic lung cancer cells whilst investigating metastasis-related molecular features. Results: We discovered that BCAT1, the key enzyme in branched-chain amino acid metabolism, is overexpressed at the protein level in metastatic lung cancer cells, as well as in metastatic tissues from lung cancer patients. Analysis of transcriptomic data available in the TCGA database revealed that increased BCAT1 transcription is associated with poor overall survival of lung cancer patients. In accord with a critical role in metastasis, shRNA-mediated knockdown of BCAT1 expression reduced migration of metastatic cells in vitro and the metastasis of these cells to distal organs in nude mice. Mechanistically, high levels of BCAT1 depleted α-ketoglutarate (α-KG) and promoted expression of SOX2, a transcription factor regulating cancer cell stemness and metastasis. Conclusion: Our findings suggest that BCAT1 plays an important role in promoting lung cancer cell metastasis, and may define a novel pathway to target as an anti-metastatic therapy.
Subject(s)
Lung Neoplasms/metabolism , Neoplasm Metastasis/genetics , Transaminases/genetics , Animals , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , China , Databases, Genetic , Female , Gene Expression/genetics , Gene Expression Regulation, Neoplastic/genetics , Humans , Lung Neoplasms/genetics , Mass Spectrometry/methods , Mice , Mice, Inbred BALB C , Mice, Nude , Proteomics/methods , SOXB1 Transcription Factors/genetics , SOXB1 Transcription Factors/metabolism , Transaminases/metabolism , Transcriptome/genetics , Xenograft Model Antitumor AssaysABSTRACT
The level of phthalate esters (PAEs) alone is not considered to be a sufficient indicator of PAE pollution due to the quick metabolism of PAEs in the biota. The primary metabolites of PAEs, monoalkyl phthalate esters (MPEs), may also be an important indicator. However, PAE metabolism has scarcely been documented in wild marine organisms. We analysed five PAEs [dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), di(2-ethylhexyl) phthalate (DEHP), and di-n-octyl phthalate (DNOP)] and their corresponding MPEs [mono-methyl phthalate (MMP), mono-ethyl phthalate (MEP), mono-n-butyl phthalate (MBP), mono-2-ethylhexyl phthalate (MEHP), and mono-n-octyl phthalate (MNOP)] in 95 wild aquatic marine samples (including fish, prawns and molluscs) collected from the Yangtze River Delta area of the East China Sea. The species-dependent distribution of these compounds was associated with the food habits, living patterns and trophic levels of the biota. Slightly higher levels of hydrophobic PAEs (DBP and DEHP, logKOW 4.27 and 7.33, respectively) were observed in fish species consuming benthic organisms or in demersal fish species, suggesting the importance of benthic organisms and sediment. Trophic dilution of both PAEs and MPEs implies further metabolic transformation at higher trophic levels. MPE tissue distributions in fish demonstrate that the highest concentrations were always observed in bile. Metabolism via the kidney and gill is a probable main way for the relatively less hydrophobic MPEs (logKOW = <4.73, from MMP to MEHP), whereas metabolism via the liver is likely the main way for the most hydrophobic MNOP (logKOW 5.22). Generally, higher detection frequencies of MPEs were observed than those of parent PAEs. Significant liner correlations were observed between the levels of short-branched (carbon atom per chain = <4) MPEs and the sum of PAEs and MPEs (n = 95, p < 0.01), demonstrating that short-branched MPEs can be used as biomarkers of exposure to quantitatively reflect parent PAE contamination in wild marine organisms.
Subject(s)
Aquatic Organisms/drug effects , Diethylhexyl Phthalate/analogs & derivatives , Environmental Monitoring/methods , Rivers/chemistry , Water Pollutants, Chemical/analysis , Animals , Aquatic Organisms/metabolism , China , Dibutyl Phthalate/analysis , Dibutyl Phthalate/chemistry , Diethylhexyl Phthalate/analysis , Diethylhexyl Phthalate/chemistry , Esters/analysis , Fishes/metabolism , Mollusca/metabolism , Phthalic Acids/analysis , Phthalic Acids/chemistry , Water Pollutants, Chemical/chemistryABSTRACT
The distributions of alkylphenol (AP) and bisphenol A (BPA) in marine organisms, including fish, prawns and molluscs, could reflect the exposure of AP and BPA directly and effectively. This paper provides the first report on the species-dependent distribution and the human health risks of four APs (4-t-octylphenol, 4-t-OP; 4-octylphenol, 4-OP;4-nonylphenol, 4-NP; 4-n-nonylphenol, 4-n-NP) and BPA in 95 wild and 88 processed marine biota samples from the East China Sea of the Yangtze River Delta area. 4-NP was the predominant compound with the highest detected concentration of 19,890.50ng/g ww. Significant pollution was observed in Taizhou, Shanghai and Ningbo. The species-dependent distribution was related to food habits, living patterns and trophic transfer. Higher residual concentrations of 4-t-OP, 4-OP, 4-n-NP and BPA were observed in fish species that consumed benthic organisms or demersal fish species, whereas 4-NP showed different results due to trophic dilution. The trophic magnification factors (TMFs) of the linear APs (4-OP and 4-n-NP) (1.22-2.93) were higher than those of the branched ones (4-t-OP and 4-NP) (0.72-0.90), indicating the relative metabolism stability of linear APs. 4-NP has the lowest TMF value of 0.72, and its trophic dilution might be observed because the branched carbon chain exhibits the lowest dispersion force compared to that of the other APs. As for the health risk, 7-year old children may be exposed to the highest health risk of 4-NP with 95th percentile values of the hazard quotient of 0.22 to 0.23; however, the risks of the other chemicals were relatively low.
Subject(s)
Aquatic Organisms/metabolism , Benzhydryl Compounds/metabolism , Endocrine Disruptors/metabolism , Environmental Monitoring , Phenols/metabolism , Water Pollutants, Chemical/metabolism , Animals , China , Humans , Seafood/statistics & numerical dataABSTRACT
This study aimed to develop a sensitive and reliable multi-residue method for the determination of trace amounts of endocrine disrupting chemicals including five phthalate esters (PAEs), five monoalky phthalate esters (MPEs), four alkylphenols (APs) and bisphenol A (BPA) in seafood. Ultrasonic liquid extraction was selected for extraction based on acetonitrile, instead of frequently-used n-hexane, due to its lower background of PAEs. Application of solid phase extraction (SPE) with primary secondary amine (PSA, 1g/6 mL) cartridge achieved the relatively low matrix effects for MPEs and BPA in seafood. To our knowledge, it is the first study reporting about simultaneous extraction and purification of PAEs, MPEs, APs and BPA in biota samples. To obtain the maximum sensitivity, both liquid chromatography tandem mass spectrometry (LC-MS/MS) and gas chromatography tandem mass spectrometry (GC-MS/MS) were applied for analysis. This method was validated and tested on fish, mollusk and prawn. Sufficient linearity was verified by Mandel's fitting test for the matrix-matched calibrations used in this study for MPEs, APs and BPA, between 0.5 ng/g and 200 ng/g or 400 ng/g. And correlation coefficients of all calibrations suppressed 0.99 for all analytes. Good recoveries were obtained, ranging from 60% to 127% for most compounds. The sensitivity was good with method detection limits (MDLs) of 0.015-2.2 ng/g wet weight (ww) for all compounds. Most MDLs are much lower than those in previous reports. The sensitive method was then applied on real fish, mollusk and prawn samples from the Yangtze River Delta sea area (China), and all the target compounds were detected with the maximum concentrations of PAEs, MPEs, APs and BPA up to 219.3 ng/g ww, 51.4 ng/g ww, 62.0 ng/g ww and 8.6 ng/g ww, respectively.