Evaluation of neurological behavior alterations and metabolic changes in mice under chronic glyphosate exposure.

Glyphosate is a widely used active ingredient in agricultural herbicides, inhibiting the biosynthesis of aromatic amino acids in plants by targeting their shikimate pathway. Our gut microbiota also facilitates the shikimate pathway, making it a vulnerable target when encountering glyphosate. Dysbiosis in the gut microbiota may impair the gut-brain axis, bringing neurological outcomes. To evaluate the neurotoxicity and biochemical changes attributed to glyphosate, we exposed mice with the reference dose (RfD) set by the U.S. EPA (1.75 mg/Kg-BW/day) and its hundred-time-equivalence (175 mg/Kg-BW/day) chronically via drinking water, then compared a series of neurobehaviors and their fecal/serum metabolomic profile against the non-exposed vehicles (n = 10/dosing group). There was little alteration in the neurobehavior, including motor activities, social approach, and conditioned fear, under glyphosate exposure. Metabolomic differences attributed to glyphosate were observed in the feces, corresponding to 68 and 29 identified metabolites with dysregulation in the higher and lower dose groups, respectively, compared to the vehicle-control. There were less alterations observed in the serum metabolome. Under 175 mg/Kg-BW/day of glyphosate exposure, the aromatic amino acids (phenylalanine, tryptophan, and tyrosine) were reduced in the feces but not in the serum of mice. We further focused on how tryptophan metabolism was dysregulated based on the pathway analysis, and identified the indole-derivatives were more altered compared to the serotonin and kynurenine derivatives. Together, we obtained a three-dimensional data set that records neurobehavioral, fecal metabolic, and serum biomolecular dynamics caused by glyphosate exposure at two different doses. Our data showed that even under the high dose of glyphosate irrelevant to human exposure, there were little evidence that supported the impairment of the gut-brain axis.

Emerging organic contaminants of liquid crystal monomers: Environmental occurrence, recycling and removal technologies, toxicities and health risks.

Liquid crystal monomers (LCMs) are a family of synthetic organic chemicals applied in the liquid crystal displays (LCDs) of various electric and electronic products (e-products). Due to their unique properties (i.e., persistence, bioaccumulative potential, and toxicity) and widespread environmental distributions, LCMs have attracted increasing attention across the world. Recent studies have focused on the source, distribution, fate, and toxicity of LCMs; however, a comprehensive review is scarce. Herein, we highlighted the persistence and bioaccumulation potential of LCMs by reviewing their physical-chemical properties. The naming rules were suggested to standardize the abbreviations regarding LCMs. The sources and occurrences of LCMs in different environmental compartments, including dust, sediment, soil, leachate, air and particulate, human serum, and biota samples, were reviewed. It is concluded that the LCMs in the environment mainly originate from the usage and disassembly of e-products with LCDs. Moreover, the review of the potential recycling and removal technologies regarding LCMs from waste LCD panels suggests that a combination of natural attenuation and physic-chemical remediation should be developed for LCMs remediations in the future. By reviewing the health risks and toxicity of LCMs, it is found that a large gap exists in their toxicity and risk to organisms. The fate and toxicity investigation of LCMs, and further investigations on the effects on the human exposure risks of LCMs to residents, especially to occupational workers, should be considered in the future.

Brain organoids are new tool for drug screening of neurological diseases.

At the level of in vitro drug screening, the development of a phenotypic analysis system with high-content screening at the core provides a strong platform to support high-throughput drug screening. There are few systematic reports on brain organoids, as a new three-dimensional in vitro model, in terms of model stability, key phenotypic fingerprint, and drug screening schemes, and particularly regarding the development of screening strategies for massive numbers of traditional Chinese medicine monomers. This paper reviews the development of brain organoids and the advantages of brain organoids over induced neurons or cells in simulated diseases. The paper also highlights the prospects from model stability, induction criteria of brain organoids, and the screening schemes of brain organoids based on the characteristics of brain organoids and the application and development of a high-content screening system.

Liquid Crystal Monomer: A Potential PPARγ Antagonist.

Liquid crystal monomers (LCMs) are a large family of artificial ingredients that have been widely used in global liquid crystal display (LCD) industries. As a major constituent in LCDs as well as the end products of e-waste dismantling, LCMs are of growing research interest with regard to their environmental occurrences and biochemical consequences. Many studies have analyzed LCMs in multiple environmental matrices, yet limited research has investigated the toxic effects upon exposure to them. In this study, we combined in silico simulation and in vitro assay validation along with omics integration analysis to achieve a comprehensive toxicity elucidation as well as a systematic mechanism interpretation of LCMs for the first time. Briefly, the high-throughput virtual screen and reporter gene assay revealed that peroxisome proliferator-activated receptor gamma (PPARγ) was significantly antagonized by certain LCMs. Besides, LCMs induced global metabolome and transcriptome dysregulation in HK2 cells. Notably, fatty acid ß-oxidation was conspicuously dysregulated, which might be mediated through multiple pathways (IL-17, TNF, and NF-kB), whereas the activation of AMPK and ligand-dependent PPARγ antagonism may play particularly important parts. This study illustrated LCMs as a potential PPARγ antagonist and explored their toxicological mode of action on the trans-omics level, which provided an insightful overview in future chemical risk assessment.

Dose-response metabolomics and pathway sensitivity to map molecular cartography of bisphenol A exposure.

In the toxicological regime, the toxicological endpoint and its dose-response relationship are two of the most prominent characters in conducting a risk assessment for chemical exposure. Systems biological methods have been used to comprehensively characterize the impact of toxicants on the biochemical pathways. However, the majority of the current studies are only based on single-dose, and limited information can be extrapolated to other doses from these experiments, regardless of the sensitivity of each endpoint. This study aims to understand the dose-response metabolite dysregulation pattern and metabolite sensitivity at the system-biological level. Here, we applied bisphenol A (BPA), an endocrine-disrupting chemical (EDC), as the model chemical. We first employed the global metabolomics method to characterize the metabolome of breast cancer cells (MCF-7) upon exposure to different doses (0, 20, 50, and 100 µM) of BPA. The dysregulated features with a clear dose-response relationship were also effectively picked up with an R-package named TOXcms. Overall, most metabolites were dysregulated by showing a significant dose-dependent behaviour. The results suggested that BPA exposure greatly perturbed purine metabolism and pyrimidine metabolism. Interestingly, most metabolites within the purine metabolism were described as a biphasic dose-response relationship. With the established dose-response relationship, we were able to fully map the metabolite cartography of BPA exposure within a wide range of concentrations and observe some unique patterns. Furthermore, an effective concentration of certain fold changes (e.g., EC+10 means the dose at which metabolite is 10% upregulated) and metabolite sensitivity were defined and introduced to this dose-response omics information. The result showed that the purine metabolism pathway is the most venerable target of BPA, which can be a potential endogenous biomarker for its exposure. Overall, this study applied the dose-response metabolomics method to fully understand the biochemical pathway disruption of BPA treatment at different doses. Both dose-response omics strategy and metabolite sensitivity analysis can be further considered and emphasized in future chemical risk assessments.

Gut microbial metabolite p-cresol alters biotransformation of bisphenol A: Enzyme competition or gene induction?

Recent studies on pharmaceuticals have revealed the direct and indirect mechanisms that link human gut microbiome to xenobiotic biotransformation. Though environmental contaminants compose a vital portion of xenobiotics and share overlapping biotransformation pathways with gut microbial metabolites, the possible interplay between gut microbiome and biotransformation of environmental contaminants remains obscure. This study utilized bisphenol A (BPA) and p-cresol as model compounds to explore whether gut microbial metabolites could affect environmental phenol metabolism on both in vitro and in vivo models. We have observed some distinct biotransformation behavior, where in vivo mouse examination using 171 & 1972 µg/kg bw p-cresol injection exhibited enhancing effect on BPA metabolism, but p-cresol was found as a strong inhibitor from 10/5 µM in a non-competitive pattern for BPA biotransformation in in vitro models of liver S9 fractions and HepG2 cell line, respectively. A further investigation revealed that the expression of biotransformation enzyme genes including Ugt1a1, Ugt2b1, or Sult1a1 of p-cresol treated mice were dynamically induced. In silico docking approach was also utilized to explore the non-competitive inhibition mechanism by estimating the binding affinity of key enzyme SULT 1A1. Overall, our results provided a novel insight into the biotransformation interaction between gut microbiome and environmental contaminants.

A comprehensive review on the analytical method, occurrence, transformation and toxicity of a reactive pollutant: BADGE.

Bisphenol A diglycidyl ether (BADGE)-based epoxy resin is one of the most widely used epoxy resins with an annual production amount of several million tons. Compared with all other legacy or emerging organic compounds, BADGE is special due to its toxicity and high reactivity in the environment. More and more studies are available on its analytical methods, occurrence, transformation and toxicity. Here, we provided a comprehensive review of the current BADGE-related studies, with focus on its production, application, available analytical methods, occurrences in the environment and human specimen, abiotic and biotic transformation, as well as the in vitro and in vivo toxicities. The available data show that BADGE and its derivatives are ubiquitous environmental chemicals and often well detected in human specimens. For their analysis, a water-free sample pretreatment should be considered to avoid hydrolysis. Additionally, their complex reactions with endogenous metabolites are areas of great interest. To date, the monitoring and further understanding of their transport and fate in the environment are still quite lacking, comparing with its analogues bisphenol A (BPA) and bisphenol S (BPS). In terms of toxicity, the summary of its current studies and Environmental Protection Agency (EPA) ToxCast toxicity database suggests BADGE might be an endocrine disruptor, though more detailed evidence is still needed to confirm this hypothesis in in vivo animal models. Future study of BADGE should focus on its metabolic transformation, reaction with protein and validation of its role as an endocrine disruptor. We believe that the elucidation of BADGEs can greatly enhance our understandings of those reactive compounds in the environment and human.

Metabolomics and In Silico Docking-Directed Discovery of Small-Molecule Enzyme Targets.

The identification of target proteins for small molecules is of great importance in drug discovery and for understanding the cellular mode of action (MOA) of toxicants. Herein, a "bottom-up" oriented target finding strategy is proposed based on the principle that the targeted enzymes can be inferred according to their phenotypic changes at the metabolome level. Meanwhile, computer-assisted in silico molecular docking analysis was performed to evaluate the binding affinities between the chemicals and the target enzymes to further rank the possible targets. In this study, triphenyl phosphate (TPhP) was used as an example to illustrate the workflow. After a comprehensive metabolome and lipidome analysis, 51 related metabolic enzymes were selected for ranking binding energies, wherein 25 proteins exhibited a higher affinity for TPhP than for their endogenous substrates. Two proteins, hydroxyacyl-coenzyme A dehydrogenase (HADH) and 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10), were further confirmed by surface phasma resonance (SPR) and isothermal titration calorimetry (ITC) analysis, displayed Kd values at low micromolar levels for TPhP. Overall, the proposed strategy has provided a feasible means for discovering enzymatic targets for the large-scale small-molecule sets, with the advantages of closely associating with the phenotype change, reducing the cost of groping, and improving the accuracy of target prediction.

System Biology-Guided Chemical Proteomics to Discover Protein Targets of Monoethylhexyl Phthalate in Regulating Cell Cycle.

Chemical proteomics methods have been used as effective tools to identify novel protein targets for small molecules. These methods have great potential to be applied as environmental toxicants to figure out their mode of action. However, these assays usually generate dozens of possible targets, making it challenging to validate the most important one. In this study, we have integrated the cellular thermal shift assay (CETSA), quantitative proteomics, metabolomics, computer-assisted docking, and target validation methods to uncover the protein targets of monoethylhexyl phthalate (MEHP). Using the mass spectrometry implementation of CETSA (MS-CETSA), we have identified 74 possible protein targets of MEHP. The Gene Ontology (GO) enrichment integration was further conducted for the target proteins, the cellular dysregulated proteins, and the metabolites, showing that cell cycle dysregulation could be one primary change due to the MEHP-induced toxicity. Flow cytometry analysis confirmed that hepatocytes were arrested at the G1 stage due to the treatment with MEHP. Subsequently, the potential protein targets were ranked by their binding energy calculated from the computer-assisted docking with MEHP. In summary, we have demonstrated the development of interactomics workflow to simplify the redundant information from multiomics data and identified novel cell cycle regulatory protein targets (CPEB4, ANAPC5, and SPOUT1) for MEHP.

A Novel Mechanism of Monoethylhexyl Phthalate in Lipid Accumulation via Inhibiting Fatty Acid Beta-Oxidation on Hepatic Cells.

Monoethylhexyl phthalate (MEHP) is one of the main active metabolites of the plasticizer di(2-ethylhexyl) phthalate. It has been known that MEHP has an impact on lipolysis; however, its mechanism on the cellular lipid metabolism remains largely unclear. Here, we first utilized global lipid profiling to fully characterize the lipid synthesis and degradation pathways upon MEHP treatment on hepatic cells. Meanwhile, we further identified the possible MEHP-targeted proteins in living cells using the cellular thermal shift assay (CETSA) method. The lipidomics results showed that there was a significant accumulation of fatty acids and other lipids in the cell. The CETSA identified 18 proteins and fatty acid ß-oxidation inhibition pathways that were significantly perturbed. MEHP's binding with selected proteins HADH and HSD17B10 was further evaluated using molecule docking, and results showed that MEHP has higher affinities as compared to endogenous substrates, which was further experimentally confirmed in the surface plasma resonance interaction assay. In summary, we found a novel mechanism for MEHP-induced lipid accumulation, which was probably due to its inhibitive effects on the enzymes in fatty acid ß-oxidation. This mechanism substantiates the public concerns on the high exposure level to plasticizers and their possible role as an obesogen.