Predicting the Metal Mixture Toxicity with a Toxicokinetic-Toxicodynamic Model Considering the Time-Dependent Adverse Outcome Pathways.

Chemicals mainly exist in ecosystems as mixtures, and understanding and predicting their effects are major challenges in ecotoxicology. While the adverse outcome pathway (AOP) and toxicokinetic-toxicodynamic (TK-TD) models show promise as mechanistic approaches in chemical risk assessment, there is still a lack of methodology to incorporate the AOP into a TK-TD model. Here, we describe a novel approach that integrates the AOP and TK-TD models to predict mixture toxicity using metal mixtures (specifically Cd-Cu) as a case study. We preliminarily constructed an AOP of the metal mixture through temporal transcriptome analysis together with confirmatory bioassays. The AOP revealed that prolonged exposure time activated more key events and adverse outcomes, indicating different modes of action over time. We selected a potential key event as a proxy for damage and used it as a measurable parameter to replace the theoretical parameter (scaled damage) in the TK-TD model. This refined model, which connects molecular responses to organism outcomes, effectively predicts Cd-Cu mixture toxicity over time and can be extended to other metal mixtures and even multicomponent mixtures. Overall, our results contribute to a better understanding of metal mixture toxicity and provide insights for integrating the AOP and TK-TD models to improve risk assessment for chemical mixtures.

Non-negligible vector effect of micro(nano)plastics on tris(1,3-dichloro-2-propyl) phosphate in zebrafish quantified by toxicokinetic model.

Micro(nano)plastics (MNPs) inevitably interact with coexisting contaminants and can act as vectors to affect their fate in organisms. However, the quantitative contribution of MNPs in the in vivo bioaccumulation and distribution of their coexisting contaminants remains unclear. Here, by selecting tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) as the typical coexisting contaminant, we quantified the contribution of MNPs to bioaccumulation and distribution of TDCIPP with toxicokinetic models. Results indicated that MNPs differentially facilitated TDCIPP bioaccumulation and distribution, and NPs slowed down TDCIPP depuration more significantly than MPs. Model analysis further revealed increasing contributions of MNPs to whole-fish TDCIPP bioaccumulation over time, with NPs (33-42%) contributing more than MPs (12-32%) at 48 h exposure. NPs contributed more than MPs to TDCIPP distribution in the liver (13-19% for MPs; 36-52% for NPs) and carcass (24-45% for MPs; 57-71% for NPs). The size-dependent vector effect might be attributed to the fact that MNPs promote contaminant transfer by damaging biofilm structure and increasing tissue membrane permeability, with NPs exerting stronger effects. This work demonstrated the effectiveness of using modeling tools to understand the relative importance of MNPs as contaminant vectors in the TK process and highlighted the higher contaminant transfer potential of NPs under combined exposure scenarios.

Near-Infrared-II In Vivo Visualization and Quantitative Tracking of Micro/Nanoplastics in Fish.

As emerging contaminants, micro/nanoplastics (MNPs) are widely present in aquatic environments and are often ingested by aquatic organisms. However, the in vivo trafficking and fate of MNPs remain largely unknown. Here, we developed near-infrared (NIR) aggregated-induced emission (AIE) fluorophore-labeled microplastics (2 µm) and nanoplastics (100 nm) as models of MNPs. This model was based on the NIR-AIE technique with strong emission at the second near-infrared (NIR-IIII) window, which overcomes the interference of autofluorescence and observation artifacts in the detection of commercial fluorescent-labeled particles. Due to its deep tissue penetration and noninvasiveness, the dynamic process of accumulation and transport of MNPs in individuals can be tracked with NIR imaging. We then directly visualized and quantified the uptake and depuration processes of MPs and NPs in zebrafish. The results showed that the MPs and NPs were mainly accumulated in the fish gut, and the distribution was heterogeneous. MPs tended to accumulate more in the fore and mid areas of the gut compared with NPs. Besides, both MPs and NPs could accumulate in large quantities locally in the gut and might cause intestinal obstruction. MNPs accumulated slowly during the initial exposure followed by rapid and sustained accumulation in gut. Based on these kinetic accumulation and depuration, we developed a refined toxicokinetic (TK) model to describe the dynamic changes in the uptake and depuration of MNPs. Overall, this study proposed a MNP model based on the NIR-AIE technique, which provided a reliable tracer technology for the visualization, tracking and quantification of MNPs in vivo.

A refined toxicokinetic model for quantifying the interaction between Cd and Cu in zebrafish larvae.

The interaction between metals is ubiquitous, but there is still a lack of quantitative models considering the interaction between metals, which leads to the deviations in predicting the joint toxicity of metals. The present study estimated the uptake rate constants (kin) and elimination rate constants (kout) and elucidated how the presence of one metal (Cu or Cd) affects the absorption and excretion of another metal (Cd or Cu) in zebrafish larvae. The results showed that Cd and Cu inhibited each other in the process of absorption and excretion by comparing separately kin and kout of Cd or Cu with the other metal Cu or Cd mixed concentrations increased, thereby affecting the Cd and Cu bioaccumulation in the zebrafish larvae. Then the interactions between Cd and Cu in the uptake and elimination processes were quantified to obtain a refined toxicokinetic model. Verification with independent experiment data showed that the refined toxicokinetic model could significantly improve the prediction of the Cd or Cu bioaccumulation in the zebrafish larvae. This study contributes to understand the toxicokinetic process of the Cd-Cu mixture in the zebrafish larvae, and the developed model could be used to predict the toxicity of the metal mixtures.

SDG26 Is Involved in Trichome Control in Arabidopsis thaliana: Affecting Phytohormones and Adjusting Accumulation of H3K27me3 on Genes Related to Trichome Growth and Development.

Plant trichomes formed by specialized epidermal cells play a role in protecting plants from biotic and abiotic stresses and can also influence the economic and ornamental value of plant products. Therefore, further studies on the molecular mechanisms of plant trichome growth and development are important for understanding trichome formation and agricultural production. SET Domain Group 26 (SDG26) is a histone lysine methyltransferase. Currently, the molecular mechanism by which SDG26 regulates the growth and development of Arabidopsis leaf trichomes is still unclear. We found that the mutant of Arabidopsis (sdg26) possessed more trichomes on its rosette leaves compared to the wild type (Col-0), and the trichome density per unit area of sdg26 is significantly higher than that of Col-0. The content of cytokinins and jasmonic acid was higher in sdg26 than in Col-0, while the content of salicylic acid was lower in sdg26 than in Col-0, which is conducive to trichome growth. By measuring the expression levels of trichome-related genes, we found that the expression of genes that positively regulate trichome growth and development were up-regulated, while the negatively regulated genes were down-regulated in sdg26. Through chromatin immunoprecipitation sequencing (ChIP-seq) analysis, we found that SDG26 can directly regulate the expression of genes related to trichome growth and development such as ZFP1, ZFP5, ZFP6, GL3, MYB23, MYC1, TT8, GL1, GIS2, IPT1, IPT3, and IPT5 by increasing the accumulation of H3K27me3 on these genes, which further affects the growth and development of trichomes. This study reveals the mechanism by which SDG26 affects the growth and development of trichomes through histone methylation. The current study provides a theoretical basis for studying the molecular mechanism of histone methylation in regulating leaf trichome growth and development and perhaps guiding the development of new crop varieties.

Elucidating the Differences in Metal Toxicity by Quantitative Adverse Outcome Pathways.

Numerous studies have reported that the toxicity differences among metals are widespread; however, little is known about the mechanism of differences in metal toxicity to aquatic organisms due to the lack of quantitative understanding of their adverse outcome pathway. Here, we investigated the effects of Cd and Cu on bioaccumulation, gene expression, physiological responses, and apical effects in zebrafish larvae. RNA sequencing was conducted to provide supplementary mechanistic information for the effects of Cd and Cu exposure. On this basis, we proposed a quantitative adverse outcome pathway (qAOP) suitable for metal risk assessment of aquatic organisms. Our work provides a mechanistic explanation for the differences in metal toxicity where the strong bioaccumulation of Cu enables the newly accumulated Cu to reach the threshold that causes different adverse effects faster than Cd in zebrafish larvae, resulting in a higher toxicity of Cu than that of Cd. Furthermore, we proposed a parameter CIT/BCF (the ratio of internal threshold concentration and bioaccumulation factor) that helps to understand the toxicity differences by combining the information of bioaccumulation and internal threshold of adverse effects. This work demonstrated that qAOP is an effective quantitative tool for understanding the toxicity mechanism and highlight the importance of toxicokinetics and toxicodynamics at different biological levels in determining the metal toxicity.

A review of interactions of microplastics and typical pollutants from toxicokinetics and toxicodynamics perspective.

The widespread microplastics (MPs) pollution has become a concerning environmental issue. The interactions between MPs and typical pollutants may change the bioaccumulation, and toxicity of pollutants, leading to high uncertainty in risk assessment. Still, significant gaps remain in the knowledge available to integrate these interactions in the perspectives of toxicokinetics (TK) and toxicodynamics (TD), which is also an essential part of quantitative toxicological research. This review systematically summarizes the interaction between MPs and typical pollutants in TK and TD processes. MPs can be acted as the vector or sink of pollutants to increase or decrease their bioaccumulation, and also may not affect their bioaccumulation due to no interaction. The adverse outcome pathway (AOP) framework enables novel approaches for determining the interaction between MPs and pollutants in the TD process. MPs can directly or indirectly enhance, reduce and not affect the toxicity of pollutants. A series of factors influencing the interaction in TK and TD processes are summarized, including MPs characteristics and exposure scenarios. TK-TD approach can quantitatively understand the interaction between MPs and pollutants based on the mechanism. Given the current knowledge gap in TK and TD processes, future perspectives on combined exposure research are proposed.

Role of Toxicokinetic and Toxicodynamic Parameters in Explaining the Sensitivity of Zebrafish Larvae to Four Metals.

Given the persistence and toxic potencies of metal contaminants in ecosystems, animals, and human beings, they are considered to be hazardous global pollutants. While the lethality of metal toxicities (e.g., LC50) can significantly vary, even within the same species, the underlying mechanisms are less well-understood. In this study, we developed a subcellular two-compartment toxicokinetic-toxicodynamic (TK-TD) model for zebrafish larvae when exposed to four metals (cadmium, lead, copper, and zinc) to reveal whether differences in metal toxicity (LC50 values) were dominated by the TK or TD processes. Results showed that the subcellular TK and TD parameters of the four metals were significantly different, and the bioconcentration factor (BCF) value of copper was higher than those of the other metals. We also found that the TD parameter internal threshold concentration (CIT) was significantly positively correlated to the LC50 values (R2 = 0.7), suggesting a dominant role of TD processes in metal toxicity. Furthermore, the combined parameter CIT/BCF for a metal-sensitive fraction (BCFMSF), which linked exposure to effects through the TK-TD approach, explained up to 89% of the variation in toxicity to the four metals. The present study suggests that the observed variation in toxicity of these four metals was mainly determined by TD processes but that TK processes should not be ignored, especially for copper.