Comparison of the Glucocorticoid Receptor Binding and Agonist Activities of Typical Glucocorticoids: Insights into Their Endocrine Disrupting Effects.

Over the past decades, the synthetic glucocorticoids (GCs) have been widely used in clinical practice and animal husbandry. Given the health hazard of these toxic residues in food, it is necessary to explore the detailed interaction mechanisms of typical GCs and their main target glucocorticoid receptor (GR). Hence, this work compared the GR binding and agonist activities of typical GCs. Fluorescence polarization assay showed that these GCs were potent ligands of GR. Their GR binding affinities were in the order of methylprednisolone>betamethasone≈prednisolone>dexamethasone, with IC50 values of 1.67, 2.94, 2.95, and 5.58 nM. Additionally, the limits of detection of dexamethasone, betamethasone, prednisolone, and methylprednisolone were 0.32, 0.14, 0.19, and 0.09 µg/kg in fluorescence polarization assay. Reporter gene assay showed that these GCs induced GR transactivation in a dose-dependent manner, confirming their GR agonist activities. Among which, dexamethasone at the concentration of 100 nM produced a maximal induction of more than 11-fold over the blank control. Molecular docking and molecular dynamics simulations suggested that hydrogen-bonding and hydrophobic interactions played an important role in stabilizing the GC-GR-LBD complexes. In summary, this work might help to understand the GR-mediated endocrine disrupting effects of typical GCs.

Mangrove forest: An important coastal ecosystem to intercept river microplastics.

The research on transportation of river microplastics (MPs) mainly focuses on the estimations of the total contents of river MPs entering the ocean, while the related transportation processes and influence factors were still largely unknown. In our study, the role of mangrove forest, a special tropical ecosystem in the estuary, on the transportations of MPs from rivers to ocean was explored. Except for the ND river with the absence of mangrove forest, the MPs collected from the water sample of the river upstream were much higher than their corresponding downstream (p < 0.05), with the interception rate of riverine MPs by mangrove forests ranging from 12.86% to 56% in dry season and 10.57%-42% in rainy season. The MPs with the characteristics of high density, larger size and regular shape were more easily intercepted. Furthermore, the combined effects of ecological indicators, the properties of mangrove and the hydrodynamic factors jointly determined the interception rates of MPs. This study provides a new perspective and data support for quantifying mangrove forests intercepting MPs in rivers as a factor of MPs retention in global rivers.

Characterization of diphenyl phthalate as an agonist for estrogen receptor: an in vitro and in silico study.

Phthalate esters (PAEs) are important pollutants in the environment, which can interfere with the endocrine system by mimicking estrogen. However, limited information is available on modulating the estrogen receptor (ER) of five PAEs including di (2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), benzyl butyl phthalate (BBP), diphenyl phthalate (DPhP) and dicyclohexyl phthalate (DCHP). This study evaluated the agonistic effects of PAEs on human ER. The cytotoxicity assay showed that there were a significant inhibition of the cell proliferation with treatment of five PAEs. Moreover, DPhP does-dependently enhanced ER-mediated transcriptional activity in the reporter gene assay. The increased expression of estrogen-responsive genes (TFF1, CTSD, and GREB1) was also observed in MCF-7 cells treated with DPhP. The result of molecular docking showed that DPhP tended to bind to the agonist conformation of ER compared with the antagonist conformation of ER, demonstrating its agonist characteristic that has been confirmed in the reporter gene assay. Thus, we found that DPhP may be evaluated as an ER agonist in vitro and it can interfere with the normal function of human ER.

Identification of dicyclohexyl phthalate as a glucocorticoid receptor antagonist by molecular docking and multiple in vitro methods.

The potential activities of phthalate esters (PAEs) that interfere with the endocrine system have been focused recently. However, information on modulating the glucocorticoid receptor (GR) of PAEs is scarce. Our aim was to evaluate the agonistic / antagonistic properties of PAEs on human GR. Luciferase reporter gene assay revealed that the tested chemicals displayed no agonistic effects but dicyclohexyl phthalate (DCHP) exerted antagonistic activity in a dose-responsive manner for GR in HeLa cells. The effects of DCHP on dexamethasone (DEX)-induced GR nuclear translocation and gene expression of glucocorticoid-responsive gene expression (G6Pase, PEPCK, FAS, GILZ and MKP-1), as well as protein expression of G6Pase and PEPCK were further examined by RT-qPCR and western blot analysis. DCHP antagonized DEX-induced GR nuclear translocation and suppressed gene expression in both mRNA and protein levels. Furthermore, the results of molecular docking and molecular dynamics simulation showed that DCHP could bind to GR and exhibited potential regulation on this target protein. Collectively, we demonstrate that DCHP may act as a GR antagonist in vitro and is considered to exert endocrine effects via human GR.

Saponins as modulators of nuclear receptors.

As plant-derived natural products, saponins have been widely applied for the dietary modification of metabolic syndrome. However, the underlying mechanisms of their preventive and therapeutic effects are still largely unclear. Nuclear receptors have been identified as potential pharmaceutical targets for treating various types of metabolic disorders. With similar structure to endogenous hormones, several saponins may serve as selective ligands for nuclear receptors. Recently, a series of saponins are proved to exert their physiological activities through binding to nuclear receptors. This review summarizes the biological and pharmacological activities of typical saponins mediated by some of the most well described nuclear receptors, including the classical steroid hormone receptors (ER, GR, MR, and AR) and the adopted orphan receptors (PPAR, LXR, FXR, and PXR).

Transcriptional response of Pseudomonas chenduensis strain MBR to cadmium toxicity.

Cadmium (Cd) contamination has resulted in serious environmental pollution and threatens human health and ecosystems. Our recent studies have demonstrated that Pseudomonas chenduensis strain MBR can decrease Cd bio-availability and reduce Cd accumulation in rice; however, the transcriptional mechanisms underlying the bacterial response during and particularly after Cd bioremediation are still unclear. In this study, we used RNA-Seq to investigate the transcriptional profiles of strain MBR during and after Cd bioremediation. During Cd bioremediation, MBR removed all Cd2+ ions in solution within 24 h, accompanied by 564 upregulated and 363 downregulated expressed genes compared with that of the control (without Cd supplementation). Specifically, under Cd stress, the upregulation of czc (czcA, czcB, and czcC) and mer (merA, merT, merC, and merP) genes enabled Cd efflux from the cytoplasm and conferred resistance of MBR to Cd toxicity. The upregulation of genes (algK, algX, and alg44) related to biofilm formation enabled Cd absorption and contributed to Cd bioremediation. After Cd bioremediation, MBR was transferred to non-Cd medium, and the genes related to histidine metabolism and flagellar assembly still showed similar expression patterns as those during bioremediation (defined as Cd legacy effects). However, the genes involved in Cd resistance and bioremediation were not influenced by Cd legacy effects. This study provides new and thorough insights into the molecular mechanisms underlying Cd bioremediation by a functional microbe. KEY POINTS: • The upregulation of czc and mer genes is responsible for MBR resistance to Cd. • The upregulation of genes related to biofilm formation contributes to Cd bioremediation. • Cd effects on genes involved in histidine metabolism and flagellar assembly are long-lasting.

Phthalate esters and dexamethasone synergistically activate glucocorticoid receptor.

This study was conducted to determine the endocrine-disrupting effects of phthalate esters (PAEs) on the glucocorticoid receptor (GR) signaling. Potential (anti)glucocorticoid activities of six typical PAEs including di (2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), dibutyl phthalate (DBP), diisobutyl phthalate (DIBP), diethyl phthalate (DEP) and dimethyl phthalate (DMP) were evaluated on human GR using cell viability assessment, reporter gene expression analysis, mRNA analysis, and molecular docking and simulation. For all tested chemicals, co-treatment of DEHP and DINP with dexamethasone (DEX) exhibited a synergistic effect on GR transactivity in the reporter assays. Such co-treatment also synergistically enhanced DEX-induced upregulation of GR mediated gene (PEPCK, FAS and MKP-1) mRNA expression in HepG2 cells and A549 cells. Molecular docking and dynamics simulations showed that hydrophobic interactions may stabilize the binding between molecules and GR. In summary, DEHP and DINP may be involved in synergistic effects via human GR, which highlight the potential endocrine-disrupting activities of PAEs as contaminants.

Dynamics and potential roles of abundant and rare subcommunities in the bioremediation of cadmium-contaminated paddy soil by Pseudomonas chenduensis.

Microbial bioremediation of heavy metal-contaminated soil is a potential technique to reduce heavy metals in crop plants. However, the dynamics and roles of the local microbiota in bioremediation of heavy metal-contaminated soil following microbial application are rarely reported. In this study, we used Pseudomonas chenduensis strain MBR for bioremediation of Cd-contaminated paddy soil and investigated its effects on the dynamics of the local soil bacterial community and Cd accumulation in rice. Cd accumulation in rice grains and roots were significantly reduced by the addition of the strain MBR. The addition of the strain MBR caused greater changes in bacterial communities in rhizosphere soil than in bulk soil. MBR enhanced the roles of microbial communities in transformation of Cd fractions, especially in rhizosphere soil. The strain MBR likely regulated abundant subcommunities more than rare subcommunities to improve Cd bioremediation, especially in rhizosphere soil. Consequently, the dynamics and functional roles of the local microbial communities differed significantly during bioremediation between abundant and rare subcommunities and between rhizosphere soil and bulk soil. This study provides new insight into the microbiota-related mechanisms underlying bioremediation.

Estrogen receptor-based multi-residue screening of bisphenol compounds in urine.

Human exposure to bisphenol compounds (BPs) has been implicated in the development of several chronic diseases. Instead of exploiting the traditional methods for determination of BPs, this work confirms that the human estrogen receptor α ligand binding domain (hERα-LBD) is a powerful recognition element that can be used to monitor multi-residue of BPs in urine samples by fluorescence polarization (FP) assay. Test parameters were optimized for the best performance. Under the optimal conditions, the IC50 values of BPs are in the range of 0.04-1.61 µg mL-1 . Recovery experiments were then performed to assess the accuracy and precision of the established method. The results detected by FP assay show good agreements with that of liquid chromatography-tandem mass spectrometry method with a fit of R2  = 0.9372 and 0.9640 for BPE and BPAP, respectively. A computational methodology, ligand-based pharmacophore model, was also employed to further explore the broad-specific of tested compounds. It was found that the two hydrogen bond acceptor features and one hydrophobic aliphatic feature were essential for the corresponding cross-reactivity results from the FP assay. All these results suggest that the established method can be successfully applied to monitor the occurrence of BPs in urine.

Multi-Residue Method for the Analysis of Stilbene Estrogens in Milk.

The rapid analysis of stilbene estrogens is crucially important in the environment, food and health sectors, but quantitation of lower detection limit for stilbene estrogens persists as a severe challenge. We herein described a homologous and sensitive fluorescence polarization (FP) assay based on estrogen receptor α ligand binding domain (ER-LBD) to monitor stilbene estrogens in milk. Under optimal conditions, the half maximal inhibitory concentrations (IC50) of the FP assay were 9.27 nM, 12.94 nM, and 22.38 nM for hexestrol, dienestrol and diethylstilbestrol, respectively. And the corresponding limits of detection (LOD) values were 2.94 nM, 2.89 nM, and 3.12 nM. Finally, the assay was applied to determine the stilbenes in milk samples where the mean recoveries ranged from 95.76% to 112.78% and the coefficients of variation (CV) below 12.00%. Furtherly, we have focused our study on high cross-reactivity phenomena by using two in silico approaches, including molecular docking analysis and topology analysis. Overall, docking results show that several residues in the hydrophobic pocket produce hydrophobic interactions with the tested drug molecules, which contribute to the stability of their binding. In this paper, we conclude that the FP method is suitable for the rapid detection of stilbenes in milk samples, requiring no expensive analytical equipment or time-consuming sample preparation. This work offers a practical approach that applies bioscience technology in food safety testing and improves analytical speed and laboratory efficiency.

Estrogenicity of halogenated bisphenol A: in vitro and in silico investigations.

The binding interactions of bisphenol A (BPA) and its halogenated derivatives (halogenated BPAs) to human estrogen receptor α ligand binding domain (hERα-LBD) was investigated using a combined in vitro and in silico approach. First, the recombinant hERα-LBD was prepared as a soluble protein in Escherichia coli BL21(DE3)pLysS. A native fluorescent phytoestrogen, coumestrol, was employed as tracer for the fluorescence polarization assay. The results of the in vitro binding assay showed that bisphenol compounds could bind to hERα-LBD as the affinity ligands. All the tested halogenated BPAs exhibited weaker receptor binding than BPA, which might be explained by the steric effect of substituents. Molecular docking studies elucidated that the halogenated BPAs adopted different conformations in the flexible hydrophobic ligand binding pocket (LBP), which is mainly dependent on their distinct halogenation patterns. The compounds with halogen substituents on the phenolic rings and on the bridging alkyl moiety acted as agonists and antagonists for hERα, respectively. Interestingly, all the compounds in the agonist conformation of hERα formed a hydrogen bond with His524, while the compounds in the antagonist conformation formed a hydrogen bond with Thr347. These docking results suggested a pivotal role of His524/Thr347 in maintaining the hERα structure in the biologically active agonist/antagonist conformation. Comparison of the calculated binding energies vs. experimental binding affinities yielded a good correlation, which might be applicable for the structure-based design of novel bisphenol compounds with reduced toxicities and for environmental risk assessment. In addition, based on hERα-LBD as a recognition element, the proposed fluorescence polarization assay may offer an alternative to chromatographic techniques for the multi-residue determination of bisphenol compounds.

In vitro and in silico assessment of the structure-dependent binding of bisphenol analogues to glucocorticoid receptor.

Widespread use of bisphenol A (BPA) and other bisphenol analogues has attracted increasing attention for their potential adverse effects. As environmental endocrine-disrupting compounds (EDCs), bisphenols (BPs) may activate a variety of nuclear receptors, including glucocorticoid receptor (GR). In this work, the binding of 11 BPs to GR was investigated by fluorescence polarization (FP) assay in combination with molecular dynamics simulations. The human glucocorticoid receptor was prepared as a soluble recombinant protein. A fluorescein-labeled dexamethasone derivative (Dex-fl) was employed as tracer. Competitive displacement of Dex-fl from GR by BPs showed that the binding affinities of bisphenol analogues were largely dependent on their characteristic functional groups. In order to further understand the relationship between BPs structures and their GR-mediated activities, molecular docking was utilized to explore the binding modes at the atomic level. The results confirmed that structural variations of bisphenol analogues contributed to different interactions of BPs with GR, potentially causing distinct toxic effects. Comparison of the calculated binding energies vs. experimental binding affinities yielded a good correlation (R 2 = 0.8266), which might be helpful for the design of environmentally benign materials with reduced toxicities. In addition, the established FP assay based on GR exhibited the potential to offer an alternative to traditional methods for the detection of bisphenols.

Insight into the role of niche concept in deciphering the ecological drivers of MPs-associated bacterial communities in mangrove forest.

Myriad inherent and variable environmental features are controlling the assembly and succession of bacterial communities colonizing on mangrove microplastics (MPs). However, the mechanisms governing mangrove MPs-associated bacterial responses to environmental changes still remain unknown. Here, we assessed the dissimilarities of MPs-associated bacterial composition, diversity and functionality as well as quantified the niche variations of each taxon on plastispheres along river-mangrove-ocean and mangrove landward-to-seaward gradients in the Beibu Gulf, China, respectively. The bacterial richness and diversity as well as the niche breadth on mangrove sedimentary MPs dramatically decreased from landward to seaward regions. Characterizing the niche variations linked the difference of ecological drivers of MPs-associated bacterial populations and functions between river-mangrove-ocean (microplastic properties) and mangrove landward-to-seaward plastispheres (sediment physicochemical properties) to the trade-offs between selective stress exerted by inherent plastic substrates and microbial competitive stress imposed by environmental conditions. Notably, Rhodococcus erythropolis was predicted to be the generalist species and closely associated to biogeochemical cycles of mangrove plastispheres. Our work provides a reliable pathway for tackling the hidden mechanisms of environmental factors driving MPs-associated microbe from perspectives of niches and highlights the spatial dynamic variations of mangrove MPs-associated bacteria.

Manufacture of Cheddar cheese using probiotic Lactobacillus plantarum K25 and its cholesterol-lowering effects in a mice model.

The probiotic adjunct Lactobacillus plantarum K25 was inoculated into milk to produce probiotic cheese. The effect of Lb. plantarum K25 on cheese composition, microbiological growth and survival during the manufacturing and ripening period, primary and secondary proteolysis during cheese ripening, and the in vivo cholesterol-lowering ability of the probiotic cheese were investigated. The results showed that the use of adjunct Lb. plantarum K25 in Cheddar cheese did not affect the cheese components including moisture, protein, fat, salt content and the pH value of cheese. During the whole ripening period, the probiotic adjunct maintained its viability, suggesting the effectiveness of Cheddar cheese as a vehicle for delivery of probiotic bacteria. No significant differences were observed in water-soluble nitrogen, 70 % ethanol-soluble nitrogen, 5 % phosphotungstic acid-soluble nitrogen, free amino acids and urea-PAGE patterns between the control and probiotic cheeses. Assessment of the in vivo cholesterol-lowering property of cheese with Lb. plantarum K25 showed that the levels of serum total cholesterol, low-density lipoprotein cholesterol and triglycerides decreased significantly, and the level of serum high-density lipoprotein cholesterol increased in mice fed with the probiotic cheese. The results indicated the potential function as a dietary item of the probiotic cheese with Lb. plantarum K25 to reduce the risk of cardiovascular diseases.

Distinct microplastics abundance variation in root-associated sediments revealed the underestimation of mangrove microplastics pollution.

Mangrove sediment is acknowledged as the critical sink of microplastics (MPs). However, the potential effect of mangrove root systems on the MPs migration in sediment remains largely unknown. Here, our study characterized the spatial distribution of MPs trapped in root hair, rhizosphere, and non-rhizosphere zones, and analyzed their correlations with physicochemical properties of sediments. The significantly increased MPs abundances toward root systems shed light on the distinct effect on the migration of MPs exerted by mangrove root systems. Partial least squares path modeling (PLS-PM) analysis revealed that pore water content and pH influenced the abundances of different MP characteristics (shape, color, size, and type) and further promoted the accumulation of MPs toward the root systems. In different mangrove areas from landward to seaward, other sediment properties (median grain size, clay content, and salinity) also controlled MP distribution. Additionally, smaller-sized MPs (<1000 µm) were more easily transported to the root systems. Our study emphasizes the importance of considering root systems effect when investigating the mechanisms of MPs distribution and migration in mangrove sediments.

Identifying the electricity-saving driving behaviors of electric bus based on trip-level electricity consumption: a machine learning approach.

Electric buses (EBs) are gaining popularity worldwide as a more sustainable and eco-friendly alternative to diesel buses (DBs). Electricity-saving driving plays a crucial role in minimizing an EB's energy consumption, subsequently leading to an extended driving range. This study proposes a machine learning-based framework for identifying electricity-saving EB driving behaviors during various driving stages, including running on road segments, entering bus stops/intersections, and exiting bus stops/intersections. The proposed random forest (RF) model effectively evaluates the energy consumption level using EB drivers' historical driving data under different scenarios. Specifically, the electricity consumption factor (ECF), as the evaluation index, is divided into three categories to determine the implicit relationship between driving behavior and energy consumption. The results indicate that the classification accuracy of RF models surpasses 90%, which highlights the effectiveness in accurately identifying energy-efficient EB driving behaviors. In addition, the Shapley additive explanations (SHAP) and partial dependency plots (PDPs) are utilized to visualize and interpret the results of RF models. A speed interval of 30-40 km/h is identified as the most energy-efficient range for EB running on a road segment. Findings from this study can be applied to targeted optimization of electricity-saving driving strategies in different driving scenarios to improve the overall efficiency and sustainability of the transportation system.

Role of mangrove forest in interception of microplastics (MPs): Challenges, progress, and prospects.

Mangroves receive microplastics (MPs) from terrestrial, marine and atmospheric sources, acting as a huge filter for environmental MPs between land and sea. Due to the high primary production and complex hydrodynamic conditions in mangroves, MPs are extensively intercepted in various ways while flowing through mangroves, leading to a long-standing but fiercely increasing MPs accumulation. However, current researches mainly focused on the occurrence, source and fate of MPs pollution in mangroves, ignoring the role of mangrove forests in the interception of MPs. Our study firstly demonstrates that mangrove ecosystems have significantly greater MPs interception capacity than their surrounding environments. Then, the current status of studies related to the interception of MPs in mangrove ecosystems is comprehensively reviewed, with the main focus on the interception process and mechanisms. At last, the most pressing shortcomings of current research are highlighted regarding the intercepted flux, interception mechanisms, retention time and ecological risks of MPs in mangrove ecosystems and the relevant future perspectives are provided. This review is expected to emphasize the critical role of mangrove forests in the interception of MPs and provide the foundational knowledge for evaluating the MPs interception effect of mangrove forests globally.

Selective enrichments for color microplastics loading of marine lipophilic phycotoxins.

Microplastics (MPs) and marine lipophilic phycotoxins (MLPs) are two classes of emerging contaminants. Together, they may exacerbate the negative impacts on nearshore marine ecosystems. Herein, the loading of 14 representative MLPs, closely related to toxin-producing algae, on MPs and their relations with colorful MPs have been explored for the first time based on both field and lab data. The objectives of our study are to explore the roles of multiple factors (waterborne MLPs and MP characteristics) in the loading of MLPs by MPs with the applications of various statistical means, and to further explore the role of the color of MP in the loading of specific MLPs through lab simulation experiments. Our results demonstrated that MPs color determined the loading of some specific MLPs on MPs and green MPs can load much more than other colorful fractions (p < 0.05). These interesting phenomena illustrated that the color effects on the loading processes of MLPs on MPs are a dynamic process, and it can be well explained by the shading effect of MP color, which may affect the growth and metabolism of the attached toxic-producing algae on MPs and hence the production of specific MLPs. Furthermore, loading of MLPs on MPs can be considered as the comprehensive physicochemical and biological processes. Our results caution us that special attention should be paid to explore the real-time dynamic color shading effects on all kinds of bio-secreted contaminants loading on MPs, and highlight the necessary to comprehensive investigate the interaction between biota, organic contaminants and MPs.

Dietary regulation of peroxisome proliferator-activated receptors in metabolic syndrome.

BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) are a class of ligand-activated nuclear transcription factors, members of the type nuclear receptor superfamily, with three subtypes, namely PPARα, PPARß/δ, and PPARγ, which play a key role in the metabolic syndrome. In the past decades, a large number of studies have shown that natural products can act by regulating metabolic pathways mediated by PPARs. PURPOSE: This work summarizes the physiological importance and clinical significance of PPARs and reviews the experimental evidence that natural products mediate metabolic syndrome via PPARs. METHODS: This study reviews relevant literature on clinical trials, epidemiology, animals, and cell cultures published in NCBI PubMed, Scopus, Web of Science, Google Scholar, and other databases from 2001 to October 2022. Search keywords were "natural product" OR "botanical" OR "phytochemical" AND "PPAR" as well as free text words. RESULTS: The modulatory involvement of PPARs in the metabolic syndrome has been supported by prior research. It has been observed that many natural products can treat metabolic syndrome by altering PPARs. The majority of currently described natural compounds are mild PPAR-selective agonists with therapeutic effects that are equivalent to synthetic medicines but less harmful adverse effects. CONCLUSION: PPAR agonists can be combined with natural products to treat and prevent metabolic syndrome. Further human investigations are required because it is unknown how natural products cause harm and how they might have negative impacts.

Quercetin ameliorates hepatic fat accumulation in high-fat diet-induced obese mice via PPARs.

As a natural pigment in food, quercetin possesses multiple biological activities and plays a crucial role in regulating metabolic syndrome. Herein, we aim to explore the potential mechanism of quercetin to ameliorate hepatic fat accumulation. In vivo experiments showed that quercetin significantly relieved inflammation response by decreasing the serum TNF-α and IL-6 levels and also improved high-fat diet-induced hepatic steatosis without other organ injuries. Quercetin can effectively reduce lipid aggregation and down-regulate the protein expression of PCK1 in HepG2 cells induced by oleic acid and palmitic acid, indicating that inhibiting gluconeogenesis leads to hepatic fat accumulation reduction. Furthermore, molecular docking results suggested that quercetin can bind to both PPARα and PPARγ, with an even more potent binding affinity than indeglitazar, a pan-agonist of PPARs. In conclusion, quercetin may regulate gluconeogenesis to ameliorate hepatic fat accumulation via targeting PPARα/γ.