High expression of YEATS2 as a predictive factor of poor prognosis in patients with hepatocellular carcinoma.

YEATS domain containing 2 (YEATS2), it may function as a proto-oncogene. This study aims to investigate if YEATS2 correlates with prognosis in hepatocellular carcinoma. The prognostic landscape of YEATS2 and its relationship with expression in hepatocellular carcinoma were deciphered with public databases, RT-qPCR and western-blot in tissue samples. The expression profiling and prognostic value of YEATS2 were explored using UALCAN, TIMER, OncoLnc database. Transcription and survival analyses of YEATS2 in hepatocellular carcinoma were investigated with cBioPortal database. The STRING database was explored to identify molecular functions and signaling pathways downstream of YEATS2. YEATS2 expression was significantly higher in hepatocellular carcinoma compared with adjacent non-malignant tissues. Promoter methylation of YEATS2 exhibited different patterns in hepatocellular carcinoma. High expression of YEATS2 was associated with poorer survival. Mechanistically, YEATS2 was involved in mediating multiple biological processes including morphogenesis and migration.

Alkylphenols disrupt estrogen homeostasis via diradical cross-coupling reactions: A novel pathway of endocrine disruption.

Estrogen, being an essential class of sex hormone, is an important target of endocrine disruption chemicals. It is well known that environmental disruptors could activate or inhibit estrogen receptors, acting as agonists or antagonists, and thus affect the circulating estrogen concentrations. Here, we report enzyme-mediated diradical cross-coupling reactions between alkylphenols (e.g., 2,4-di-tert-butylphenol [DBP], 4-nonylphenol [4-NP], and 4-tert-octylphenol [4-t-OP]) and estrogens (e.g., estradiol [E2]) that generate coupling metabolites and disrupt estrogen homeostasis. Among the phenolic xenobiotics, the screening of metabolic products revealed that alkylphenols had the highest reaction activities and generated coupling metabolites with high abundances (DBP-O-E2, 4-t-OP-O-E2, and 4-NP-O-E2). The coupling reactions were catalyzed by cytochrome P450 3A4 (CYP3A4) and verified by the detection of the coupling products in general populations. In vitro and in vivo exposures together with CYP3A4 inhibition demonstrated that cross-coupling reactions of phenols and E2 significantly reduced the normal levels of E2. We further established a unique spin-trapping-based high-throughput screening method to show the existence of diradicals in the coupling reaction. Density functional theory calculations revealed that spin aromatic delocalization was the fundamental cause of the high rebound barrier and sufficient lifetime of phenoxy radicals that enabled phenolic cross-coupling triggered by cytochrome P450. The identified mechanistic details for diradical cross-coupling reactions provide a novel pathway for phenolic chemicals to disrupt estrogen homeostasis.

Environmental biotransformation mechanisms by flavin-dependent monooxygenase: A computational study.

The enzyme-catalyzed metabolic biotransformation of xenobiotics plays a significant role in toxicology evolution and subsequently environmental health risk assessment. Recent studies noted that the phase I human flavin-dependent monooxygenase (e.g., FMO3) can catalyze xenobiotics into more toxic metabolites. However, details of the metabolic mechanisms are insufficient. To fill the mechanism in the gaps, the systemic density functional theory calculations were performed to elucidate diverse FMO-catalyzed oxidation reactions toward environmental pollutants, including denitrification (e.g., nitrophenol), N-oxidation (e.g., nicotine), desulfurization (e.g., fonofos), and dehalogenation (e.g., pentachlorophenol). Similar to the active center compound 0 of cytochrome P450, FMO mainly catalyzed reactions with the structure of the tricyclic isoalloxazine C-4a-hydroperoxide (FADHOOH). As will be shown, FMO-catalyzed pathways are more favorable with a concerted than stepwise mechanism; Deprotonation is necessary to initiate the oxidation reactions for phenolic substrates; The regioselectivity of nicotine by FMO prefers the N-oxidation other than N-demethylation pathway; Formation of the P-S-O triangle ring is the key step for desulfurization of fonofos by FMO. We envision that these fundamental mechanisms catalyzed by FMO with a computational method can be extended to other xenobiotics of similar structures, which may aid the high-throughput screening and provide theoretical predictions in the future.

Potential health benefits of lowering gas production and bifidogenic effect of the blends of polydextrose with inulin in a human gut model.

Polydextrose is a nutrient supplement, which is widely applied in the food industry. The use of polydextrose in combination with prebiotics and probiotics has recently increased, whereas the fermentation properties of its blend have not yet been fully revealed. We evaluated the metabolic profile of polydextrose, inulin, and their blends by a batch in vitro fermentation of fifteen human fecal inocula. After 24 h of fermentation, polydextrose increased the production of gas, ammonia, and several short chain fatty acids, including propionate and butyrate, when compared to its blends, inulin, and fructo-oligosaccharides. Furthermore, polydextrose had the slowest degradation rate of all the carbohydrates tested, consistent with its partial fermentation in the distal colon. The 16S rRNA gene sequencing analysis of the gut microbiome exhibited significantly increased relative abundance of Clostridium_XVIII, Megamonas, Mitsuokella, and Erysipelotrichaceae_incertae_sedis in polydextrose compared to other carbohydrates. On the other hand, the blends of polydextrose and inulin (1:1 or 2:1) showed reduced gas production and similar bifidogenicity to inulin alone. The blends not only had similar alpha-diversity and PCoA to inulin but also had a similar abundance of beneficial bacteria, such as Faecalibacterium and Roseburia, suggesting potential health benefits. Also their low gas production was likely due to the abundance of Faecalibacterium and Anaerostipes, which were negatively correlated with gas production. Additionally, our in vitro fermentation model shows advantages in the large-scale assessment of fermentation performance.

In Silico simulation of Cytochrome P450-Mediated metabolism of aromatic amines: A case study of N-Hydroxylation.

Aromatic amines, the widely used raw materials in industry, cause long-term exposure to human bodies. They can be metabolized by cytochrome P450 enzymes to form active electrophilic compounds, which will potentially react with nucleophilic DNA to exert carcinogenesis. The short lifetime and versatility of the oxidant (a high-valent iron (IV)-oxo species, compound I) of P450 enzymes prompts us to use theoretical methods to investigate the metabolism of aromatic amines. In this work, the density functional theory (DFT) has been employed to simulate the hydroxylation metabolism through H-abstraction and to calculate the activation energy of this reaction for 28 aromatic amines. The results indicate that the steric effects, inductive effects and conjugative effects greatly contribute to the metabolism activity of the chemicals. The further correlation reveals that the dissociation energy of -NH2 (BDEN-H) can successfully predict the time-consuming calculated activation energy (R2 for aromatic and heteroaromatic amines are 0.93 and 0.86, respectively), so BDEN-H can be taken as a key parameter to characterize the relative stability of aromatic amines in P450 enzymes and further to quickly assess their potential toxicity. The validation results prove such relationship has good statistical performance (qcv2 for aromatic and heteroaromatic amines are 0.95 and 0.90, respectively) and can be used to other aromatic amines in the application domain, greatly reducing computational cost and providing useful support for experimental research.

Exposure patterns, chemical structural signatures, and health risks of pesticides in breast milk: A multicenter study in China.

China is the world's largest pesticide user. These chemicals are bioaccumulative in the human body, and eventually could be transferred from the mother to the fetus/infant via placental and breastfeeding transport, which might pose developmental deficiency risks. In this study, human biomonitoring of legacy pesticides was conducted in three Chinese cities using 60 breast milk samples. The patterns, chemical structural signatures, and the estimated daily intake of pesticides were assessed. The median concentration of HCB (57.8 ng g-1 lw, Interquartile range: 28.5-76.9 ng g-1 lw) was the highest among all pesticides, whereas the HCHs, DDXs, TCVP, and heptachlor were also detected. A significantly different pattern of pesticides was found among three sampling cities: the Mianyang cases were mostly DDXs oriented while the Wuhan and Hangzhou cases were under HCB, HCHs, TCVP, and heptachlor influences. Maternal age and pre-pregnancy BMI were found to be the influencing factors for the pesticides in the breast milk, and dietary preferences were an important factor in the exposure scenario. Chemical structural signatures indicated that for HCHs and DDXs the exposure was mostly historical, while the lindane and dicofol exposure may exist among the volunteering mothers. The EF for chiral pesticides did not deviate significantly from the racemic value. The risk from breastfeeding was negligible according to the Chinese and UN standard, while some cases from Hangzhou and Wuhan exceeded the Canadian restrictions. Thus, the adverse health effects of chemical exposure by dietary intake for infants need to be closely monitored in future studies.

Transplacental transfer mechanism of organochlorine pesticides: An in vitro transcellular transport study.

Recent studies show that, even after being banned for agricultural applications for over 30 years, organochlorine pesticides (OCPs), including hexachlorocyclohexanes (HCHs) and dichlorodiphenyltrichloroethane (DDT) and its metabolites (DDXs), can still be found in various biological matrices and pose a potential hazard to the fetus in the womb. This study aimed to investigate the possible transplacental transfer mechanism of OCPs using an in vitro placental model. The results showed that for HCHs and DDXs, the placenta had a potential protection mechanism for the fetus by having higher efflux than intake active transport efficiency to transfer the xenobiotic out of the fetal circulation. No enantiomer-specific transport was observed for the chiral OCPs in vitro, hints simple diffusion played the major role in the transplacental transfer. Metabolic and transporter inhibitors were applied in the transepithelial transport experiment to evaluate the role that major transporting protein played in the active efflux process. The ATP production inhibitors were observed to have significant inhibition on transfer, proving the hypothesis that active transport participates in the transplacental transport of OCPs in humans. Multiple transporters contributed simultaneously in the active transport for the OCPs. In this study, we could confirm that the transplacental transfer of OCPs is a combination of simple diffusion and active transport. ATP-binding cassette (ABC) superfamily transporters on the placenta contribute in the active transport. These findings could improve the understanding of the mechanisms of transplacental transfer of the OCPs.

Supporting dataset and methods for Transplacental Transfer of Organochlorine Pesticides: Concentration Ratio and Chiral Properties.

The dataset and methods provided in this article supports "Transplacental Transfer of Organochlorine Pesticides: Concentration Ratio and Chiral Properties" [1]. The supplementary data were as follows: 1) the information on pretreatment and instrumental methods.2) the data for concentration in the maternal serum, cord serum and placenta samples and data interpretation. 3) the correlation between the influence factors and the log-transformed concentrations of the chemicals samples. 4) the dataset for transplacental transfer ratio of the OCPs and correlations with influencing factors.

Enantioseparation of chiral perfluorooctane sulfonate (PFOS) by supercritical fluid chromatography (SFC): Effects of the chromatographic conditions and separation mechanism.

Perfluorooctane sulfonate (PFOS) is one of the most frequently detected perfluoroalkyl substances in environmental and human samples. Previous studies have shown that nonracemic PFOS in biological samples can be used as a marker of PFOS exposure sources. In recent years, supercritical fluid chromatography (SFC) has emerged as a powerful method to separate chiral compounds. In this study, a method of perfluoro-1-methylheptane sulfonate (1 m-PFOS) enantioseparation by SFC was established. The optimal separation was obtained using a Chiralpak QN-AX column with CO2 /2-propanol (70/30, v/v) as the mobile phase with a flow rate of 1 mL/min, column temperature was 32°C, and BPR pressure was 1800 psi. The resolution (Rs) and retention time were 0.88 and 130 minutes, respectively. This method is more economic and greener than HPLC. Modifier pH and column temperature were determined to be significant factors of SFC chiral separation. Modifier pH is negatively correlated with the retention factors and Rs. Adsorption thermodynamics were used to explain the influence of temperature change, and it was concluded that the transfer of two enantiomers from the mobile phase to the stationary phase is enthalpy-driven. Enantioseparation of 1 m-PFOS by SFC follows the same rules of ion exchange as those for the chiral separation by HPLC.

Transplacental transfer of organochlorine pesticides: Concentration ratio and chiral properties.

Currently, there is limited information about the mechanism of the human transplacental transfer for organochlorine pesticides (OCPs). This study aimed to evaluate the transplacental transfer of OCPs to better understand the influencing factors of exposure and transplacental efficiency. The study involved quantitative determination of OCPs and the enantiomer fraction (EF) of chiral OCPs in pregnant women from Wuhan, China. The results indicate that the exposure levels of OCPs varied in the order: maternal serum > cord serum > placenta. Chiral contaminants, such as α-HCH, o,p'-DDD and o,p'-DDT, were non-racemic in the three biological matrices, wherein EFα-HCH < 0.5, EFo,p'-DDD < 0.5, EFo,p'-DDT > 0.5. For HCHs, the concentration ratio between cord serum and maternal serum (Rcm) <1, while for DDXs, the Rcm ≈ 1, indicating that the transport efficiency of different pollutants is related to the physicochemical properties. These results showed that placenta seems to be a more efficient barrier for ß-HCH than for p,p'-DDE. The concentration ratios across placenta significantly lower than 1 and the enantiomeric selective transfer imply that some OCPs may have more complicated maternal-fetus transfer mechanisms, involving both simple diffusion and active transport. To the best of our knowledge, this is the first study to investigate the transfer of OCPs and their enantiomer fractions across placenta. These findings could expand the database of chemical exposure in biological matrices and improve the understanding of the mechanisms of transplacental transfer of OCPs.

A proposed method of enantioselectivity analysis for residual chiral PCBs in gas chromatography.

Polychlorinated biphenyls (PCBs) are harmful and persistent organic pollutants. The influence of chiral PCBs acts mainly on the different enantiomer fraction, bioaccumulation and even degradation in environmental media. Solvents and temperatures existed almost everywhere during the analysis of extraction, purification, concentration and detection, which was often underestimated in previous studies. In our study, the configuration stability of the chiral PCBs was examined from solvent and temperature aspects. Transformation phenomena for the analytic stereoisomer monomers of PCB45, PCB95, and PCB149 affected by temperature were observed. We demonstrated that higher inlet temperatures could increase the sensibility for the low-concentration environmental samples, resulting in isomerization of chiral PCBs. Real rice samples were used to verify our analysis method. Combined with density functional theory, we verified the mechanism of isomer conversion with various numbers and sites of the -Cl substituent. PCBs with tetra-ortho substituents (2, 2', 6, 6') were relatively stable and showed the highest rotational barriers (Ea) at approximate 240 kJ mol-1. Others with trio-ortho substituents (2, 2', 6/6') showed Ea from 170 to 190 kJ mol-1, whose enantiomeric fractions would be affected by temperature during the analysis process for environmental detection. The method we developed was a promising means to understand the mechanism of isomerization and to predict stabilities of chiral PCBs.

Enantioseparation of napropamide by supercritical fluid chromatography: Effects of the chromatographic conditions and separation mechanism.

Supercritical fluid chromatography (SFC) is already used for enantioseparation in the pharmaceutical industry, but it is rarely used for the separation of chiral pesticides. Comparing with high performence liquid chromatography, SFC uses much more environmnetal friendly and economic mobile phase, supercritical CO2 . In our work, the enantioseparation of an amide herbicide, napropamide, using three different polysaccharide-type chiral stationary phases (CSPs) in SFC was investigated. By studying the effect of different CSPs, organic modifiers, temperature, back-pressure regulator pressures, and flow rates for the enantioseparation of napropamide, we established a rapid and green method for enantioseparation that takes less than 2 minutes: The column was CEL2, the mobile phase was CO2 with 20% 2-propanol, and the flow rate was 2.0 mL/min. We found that CEL2 demonstrated the strongest resolution capability. Acetonitrile was favored over alcoholic solvents when the CSP was amylose and 2-propanol was the best choice when using cellulose. When the concentration of the modifiers or the flow rate was decreased, resolutions and analysis times increased concurrently. The temperature and back-pressure regulator pressure exhibited only minor influences on the resolution and analysis time of the napropamide enantioseparations with these chiral columns. The molecular docking analysis provided a deeper insight into the interactions between the enantiomers and the CSPs at the atomic level and partly explained the reason for the different elution orders using the different chiral columns.

Activity, toxicity, molecular docking, and environmental effects of three imidazolinone herbicides enantiomers.

All imidazolinone (IMI) herbicides are chiral consisting of two enantiomers; however, studies on the enantioselectivities of their interactions are limited. This study is a systematic assessment of the enantiomers and racemates of IMI herbicides, including semi-preparation and determination of absolute configurations, stereoselective bioactivity toward target organisms (Echinochloa crus-galli and Microcystis aeruginosa), and toxicity toward Michigan Cancer Foundation-7 (MCF-7) cells. R-imidazolinones were found to be more active than S-IMIs in the inhibition of target organisms, and neither enantiomer had estrogenic activity. Moreover, the relationship between the molecular structures and the efficiency of target growth inhibition by the IMI herbicides was investigated. Molecular modeling provided the rational structural basis for the enantioselectivity of the acetohydroxyacid synthase (AHAS) activity of the IMIs. These findings encourage the application of enantiopure R-IMI herbicides to capitalize on their advantages over the racemates.