Dual-responsive 3D DNA nanomachines cascaded hybridization chain reactions for novel self-powered flexible microRNA-detecting platform.

The microRNA-21 is closely related to chromatin remodeling and epigenetic regulation. In this work, an efficient double-response 3D DNA nanomachine (DRDN) was assembled by co-immobilizing two different lengths of hairpin DNA on the surface of gold nanoparticles (AuNPs) to capture microRNA-21 (miRNA-21), recycle miRNA-21, and trigger hybridization chain reactions (HCR). This work reports the fabrication of a laser-scribed graphene (LSG) electrode with excellent flexibility and electrical conductivity by laser-scribing commercial polyimide films (PI). The as-proposed self-powered biosensing platform presents significantly increased instantaneous current to in real-time monitor miRNA-21 by a capacitor. The biosensing platform exhibited highly sensitive detection of miRNA-21 with a detection limit of 0.142 fM in the range of 0.5 fM to 1 × 104 fM, and demonstrated high efficiency in the analysis of the tumor markers.

Prevention of methylmercury-triggered ROS-mediated impairment of embryo development by co-culture with adult adipose-derived mesenchymal stem cells.

Methylmercury (MeHg) is a potent toxin that exerts deleterious effects on human health via environmental contamination. Significant effects of MeHg on neuronal development in embryogenesis have been reported. Recently, our group demonstrated that MeHg exerts toxic effects on pre- and post-implantation embryonic development processes from zygote to blastocyst stage. Our results showed that MeHg impairs embryo development by induction of apoptosis through reactive oxygen species (ROS) generation that triggers caspase-3 cleavage and activation, which, in turn, stimulates p21-activated kinase 2 (PAK2) activity. Importantly, ROS were identified as a key upstream regulator of apoptotic events in MeHg-treated blastocysts. Data from the current study further confirmed that MeHg exerts hazardous effects on cell proliferation, apoptosis, implantation, and pre- and post-implantation embryo development. Notably, MeHg-induced injury was markedly prevented by co-culture with adipose-derived mesenchymal stem cells (ADMSCs) in vitro. Furthermore, ADMSC injection significantly reduced MeHg-mediated deleterious effects on embryo, placenta, and fetal development in vivo. Further investigation of the regulatory mechanisms by which co-cultured ADMSCs could prevent MeHg-induced impairment of embryo development revealed that ADMSCs effectively reduced ROS generation and its subsequent downstream apoptotic events, including loss of mitochondrial membrane potential and activation of caspase-3 and PAK2. The collective findings indicate that co-culture with mesenchymal stem cells (MSCs) or utilization of MSC-derived cell-conditioned medium offers an effective potential therapeutic strategy to prevent impairment of embryo development by MeHg.

Novel hollow MoS2@C@Cu2S heterostructures for high zinc storage performance.

Heterostructured materials have great potential as cathodes for zinc-ion batteries (ZIBs) because of their fast Zn2+ transport channels. Herein, hollow MoS2@C@Cu2S heterostructures are innovatively constructed using a template-engaged method. The carbon layer improves the electrical conductivity, provides a high in situ growth area, and effectively restricts volume expansion during the recycling process. MoS2 nanosheets are grown on the surfaces of hollow C@Cu2S nanocubes using the in situ template method, further expanding the specific surface area and exposing more active sites to enhance the electrical conductivity. As expected, an admirable reversible capacity of 197.2 mA h g-1 can be maintained after 1000 cycles with a coulombic efficiency of 91.1%. Therefore, we firmly believe that this work points the way forward for high-performance materials design and energy storage systems.

Self-Powered Biosensing System with Multivariate Signal Amplification for Real-Time Amplified Detection of PDGF-BB.

A self-powered biosensing system with multivariate signal amplification is designed for the ultrasensitive, highly efficient, rapid-response, and real-time detection of platelet-derived growth factor-BB (PDGF-BB). The biosensing system is composed of enzymatic biofuel cells (EBFCs), a capacitor, a digital multimeter (DMM), and a computer. Using the hybridization chain reaction (HCR), a few single DNA chains are transformed into abundant double-helix chains, which stimulates the reduction of [Ru(NH3)6]3+ to [Ru(NH3)6]2+ by electrostatic interaction, corresponding to the "on" state for HCR. As a result, the open-circuit voltage (EOCV) is significantly increased in this self-powered biosensing system. When PDGF-BB is present, a binding interaction between the target and the aptamer, i.e., PDGF-BB/Apt, corresponding to the "off" state for HCR, results in a decrease of EOCV. The PDGF-BB concentration is inversely proportional to EOCV, allowing readable, effective, and precise real-time detection of PDGF-BB. The detection limit of the biosensing system is 0.031 pg/mL (S/N = 3). This strategy provides a promising and powerful tool for the early clinical diagnosis of related colorectal cancer markers.

Novel Ultralow-Potential Electrochemiluminescence Aptasensor for the Highly Sensitive Detection of Zearalenone Using a Resonance Energy Transfer System.

An ultralow-potential electrochemiluminescence (ECL) aptasensor has been designed for zearalenone (ZEN) assay based on a resonance energy transfer (RET) system with SnS2 QDs/g-C3N4 as a novel luminophore and CuO/NH2-UiO-66 as a dual-quencher. SnS2 QDs were loaded onto g-C3N4 nanosheets and enhanced the ECL luminescence via strong synergistic effects under an ultralow potential. The UV-vis absorption spectrum of CuO/NH2-UiO-66 exhibits considerable overlap with the ECL emission spectrum of SnS2 QDs/g-C3N4, an important consideration for the RET process. In order to stimulate RET, the ZEN aptamer and complementary DNA are introduced for conjugation between the donor and the acceptor. With the binding interaction between ZEN by its aptamer, CuO/NH2-UiO-66 is removed from the electrode surface, resulting in the inhibition of the RET system and an increase in the ECL signal. Under optimal conditions, the as-prepared aptasensor quantified ZEN from 0.5 µg·mL-1 to 0.1 fg·mL-1 with a low limit of detection of 0.085 fg·mL-1, and it exhibited good stability, excellent specificity, high reproducibility, and desirable practicality. The sensing strategy provides a method for mycotoxins assay to monitor food safety.

Self-Powered Biosensor Based on DNA Walkers for Ultrasensitive MicroRNA Detection.

A novel self-powered biosensor is fabricated for ultrasensitive microRNA-21 (miRNA-21) detection, which includes an enzymatic biofuel cell (EBFC), DNA walkers, a digital multimeter (DMM), and a capacitor. As a novel strategy for signal amplification, DNA walkers are designed in the cathode, while the capacitor stores electrochemical energy from the EBFC to further boost the instantaneous current displayed by the DMM. When miRNA-21 is present, the DNA walkers are provoked to walk from as-opened hairpin structures to other hairpin structures, generating double-strand DNA structures, which stimulate [Ru(NH3)6]3+ to be adsorbed on the cathode surface by electrostatic interaction. Afterward, [Ru(NH3)6]3+ is reduced to [Ru(NH3)6]2+, and the open circuit voltage (EOCV) is significantly increased. Depending on the approach of signal amplification from DNA walkers, this biosensor displays an ultrasensitive assay toward miRNA-21 in the range of 0.5 to 104 fM, with a detection limit of 0.15 fM. In addition, this self-powered biosensor displays high selectivity for miRNA-21 assay in human serum samples.

Role of activated p21-activated kinase 2 in methylmercury-induced embryotoxic effects on mouse blastocysts.

Methylmercury (MeHg), a biotransformation product derived from mercury or inorganic mercury compounds in waterways, is a potent toxin that exerts hazardous effects on human health via environmental contamination. Previous studies have reported MeHg-induced impairment of nerve development in embryogenesis and placental development. However, the potential deleterious effects and regulatory mechanisms of action of MeHg on pre- and post-implantation embryo development are yet to be established. Experiments from the current study clearly demonstrate that MeHg exerts toxic effects on early embryonic development processes, including the zygote to blastocyst stage. Induction of apoptosis and decrease in embryo cell number were clearly detected in MeHg-treated blastocysts. Additionally, intracellular reactive oxygen species (ROS) generation and activation of caspase-3 and p21-activated protein kinase 2 (PAK2) were observed in MeHg-treated blastocysts. Importantly, prevention of ROS generation by pre-treatment with Trolox, a potent antioxidant, significantly attenuated MeHg-triggered caspase-3 and PAK2 activation as well as apoptosis. Notably, the downregulation of PAK2 via transfection of specifically targeted siRNA (siPAK2) led to marked attenuation of PAK2 activity and apoptosis and the deleterious effects of MeHg on embryonic development in blastocysts. Our findings strongly suggest that ROS serve as an important upstream regulator to trigger the activation of caspase-3, which further cleaves and activates PAK2 in MeHg-treated blastocysts. Activated PAK2 promotes apoptotic processes that, in turn, cause sequent impairment of embryonic and fetal development.

Self-Powered Biosensor for a Highly Efficient and Ultrasensitive Dual-Biomarker Assay.

A dual-biomarker, self-powered biosensor was fabricated for the ultrasensitive detection of microRNA-21 (miRNA-21) and miRNA-155 based on enzymatic biofuel cells (EBFCs), catalytic hairpin assembly (CHA), and DNA hybridization chain reaction (HCR), with a capacitor and digital multimeter (DMM). In the presence of miRNA-21, the CHA and HCR are triggered and lead to the generation of a double-helix chain, which stimulates [Ru(NH3)6]3+ to move to the biocathode surface due to electrostatic interaction. Subsequently, the biocathode obtains electrons from the bioanode and reduces [Ru(NH3)6]3+ to [Ru(NH3)6]2+, which significantly increases the open-circuit voltage (E1OCV). When miRNA-155 is present, CHA and HCR cannot be completed, resulting in a low E2OCV. The self-powered biosensor allows for the simultaneous ultrasensitive detection of miRNA-21 and miRNA-155 with detection limits of 0.15 and 0.66 fM, respectively. Moreover, this self-powered biosensor exhibits the highly sensitive detection for miRNA-21 and miRNA-155 assay in human serum samples.

The self-powered electrochemical biosensing platform with multi-amplification strategy for ultrasensitive detection of microRNA-155.

A self-powered biosensor (SPB) was constructed for the ultra-sensitive detection of microRNA-155 (miR-155) by combining a capacitor/enzymatic biofuel cell (EBFC), a strategy of rolling circle amplification (RCA) and a digital multimeter (DMM). The experimental results show that the sensitivity of the assembled EBFC-SPB can reach 15.85 µA/pM with the action of matching capacitor, which is 513% of that without capacitor (3.09 µA/pM). This achieves the first signal amplification. Furthermore, when the target miR-155 triggers RCA, electrons are continuous generated and flow to the biocathode through the external circuit to catalyze the reduction of oxygen and release [Ru(NH3)6]3+ electron acceptor. This achieves the second signal amplification. Finally, DMM is used to convert the signal into instantaneous current and amplify it for real-time reading. This achieves the third signal amplification. Therefore, the limit of detection (LOD) of the developed biosensor is as low as 0.17 fM (S/N = 3), and the linear range is between 0.5 fM and 10,000 fM, indicating that the EBFC-SPB has a broad application prospect for cancer marker of miR-155 with ultrasensitive detection.

Vertical aerosol data assimilation technology and application based on satellite and ground lidar: A review and outlook.

Observations and numerical models are mainly used to investigate the spatiotemporal distribution and vertical structure characteristics of aerosols to understand aerosol pollution and its effects. However, the limitations of observations and the uncertainties of numerical models bias aerosol calculations and predictions. Data assimilation combines observations and numerical models to improve the accuracy of the initial, analytical fields of models and promote the development of atmospheric aerosol pollution research. Numerous studies have been conducted to integrate multi-source data, such as aerosol optical depth and aerosol extinction coefficient profile, into various chemical transport models using various data assimilation algorithms and have achieved good assimilation results. The definition of data assimilation and the main algorithms will be briefly presented, and the progress of aerosol assimilation according to two types of aerosol data, namely, aerosol optical depth and extinction coefficient, will be presented. The application of vertical aerosol data assimilation, as well as the future trends and challenges of aerosol data assimilation, will be further analysed.

Real-time multiple signal amplification self-powered biosensing platform for ultrasensitive detection of MicroRNA.

A real-time self-powered biosensor is designed for ultrasensitive detection of microRNA-21 based on electrochemical energy device capacitor and target-induced recycling double amplification strategy, which greatly improves the output signal by converting a small number of targets into two glucose oxidase labeled output strand DNAs, and the squeezed-out output strand is recycled by the cathode to fix more signal [Ru(NH3)6]3+ to further improve the detection signal. A digital multimeter (DMM) is connected to computer for real-time displaying the output signal of the self-powered biosensing system, which improves the accuracy of the sensing platform. The sensitivity of the proposed biosensor is 116.15 µA/pM for target microRNA-21, which is 32.26 times higher than that of pure EBFC (3.6 µA/pM). The target concentration is proportional to the open-circuit voltage value in a wide linear range of 0.1-10000 fM with a low detection limit of 0.04 fM (S/N = 3). The method shows high sensitivity and excellent selectivity, and can be applied to detect tumor marker microRNA-21 in biological matrix.

Variations and drivers of aerosol vertical characterization after clean air policy in China based on 7-years consecutive observations.

Understanding the aerosol vertical characterization is of great importance to both climate and atmospheric environment. This study investigated the variations of aerosol profiles over eight regions of interest in China after clean air policy (2013-2019) and discussed the drivers of the vertical aerosol structure, using observations from active satellite measurements (CALIPSO). From the annual variation, the amplitude of extinction coefficient profiles showed a decreasing trend with fluctuations, and the maximum was 0.21 km-1 in Beijing-Tianjin-Hebei (JJJ). For regions suffered from air pollution, the variation was greatest below 0.45 km, while it was between 1-1.5 km for Sichuan Basin. The correlation coefficient between the relative humidity (RH) and the extinction coefficient indicated that the increase of RH inhibited the decrease of the extinction coefficient in the Yangtze River Delta. In most regions, the main aerosol subtypes were polluted dust and polluted continental, but they were coarser in JJJ and North West. The frequency of concurrency of dust and polluted dust aerosols decreased in JJJ, but polluted continental aerosols occurred more frequently. Further, the aerosol extinction coefficient profiles under different pollution conditions showed that it changed most during heavy pollution periods in JJJ, especially in 2017, with a significant aerosol loading between ∼700 and 1200 m. The atmospheric reanalysis data revealed that the weak convergence at low level and the divergence at high level supported the upward transport of aerosols in 2017. Overall, the differences in divergence allocation, RH, and wind filed were the main meteorological drivers.

Role of caspase-3-cleaved/activated PAK2 in brusatol-triggered apoptosis of human lung cancer A549 cells.

Brusatol, a major quassinoid extract of Bruceae fructus, is an important bioactive component with antineoplastic capacity. Several beneficial pharmacological and biological properties of brusatol have been uncovered to date, including anti-inflammatory, anticolitis, antimalarial, and anticancer activities. To confer anticancer benefits, brusatol is reported to effectively inhibit the Nrf2-mediated antioxidant response and trigger apoptotic signaling. In this study, we investigated the regulatory mechanisms underlying apoptotic processes in brusatol-treated A549 cells in detail. Our experiments showed that brusatol induces cell death through intracellular ROS-triggered mitochondria-dependent apoptotic events and does not involve necrosis. Mechanistically, p21-activated protein kinase 2 (PAK2) was cleaved by caspase-3 to generate an activated p34 fragment involved in brusatol-induced apoptosis of A549 cells. Notably, PAK2 knockdown led to downregulation of caspase-3-mediated PAK2 activity, in turn, effectively attenuating brusatol-induced apoptosis, highlighting a crucial role of caspase-3-activated PAK2 in this process. Moreover, knockdown of PAK2 resulted in significant inhibition of c-Jun N-terminal kinase (JNK) activity in brusatol-treated A549 cells, clearly suggesting that JNK serves as a downstream substrate of caspase-3-cleaved/activated PAK2 in the apoptotic cascade. SP600125, a specific JNK inhibitor, significantly suppressed brusatol-induced JNK activity but only partially prevented apoptosis, implying that JNK serves as only one of a number of substrates for PAK2 in the brusatol-triggered apoptotic cascade. Based on the collective results, we propose a signaling cascade model for brusatol-induced apoptosis in human A549 cells involving ROS, caspases, PAK2, and JNK.

Low-dose silver nanoparticles plus methyl mercury exert embryotoxic effects on mouse blastocysts via endoplasmic reticulum stress and mitochondrial apoptosis.

The health and environmental impacts of the increasing commercial use of silver nanoparticles (AgNPs) are a growing concern. Methyl mercury (MeHg) is a potent toxin that biotransforms from mercury or inorganic mercury compounds in waterways and causes dangerous environmental contamination. However, the potential interactions and combined effects of AgNPs and MeHg are yet to be established. In the current study, we showed that low/non-embryotoxic doses of AgNPs and MeHg interact synergistically to induce embryotoxicity and further explored the underlying mechanisms affecting mouse embryo development. Notably, co-treatment with noncytotoxic concentrations of AgNPs (10 µM) and MeHg (0.1 µM) triggered apoptotic processes and embryotoxicity in mouse blastocysts and evoked intracellular reactive oxygen species (ROS) generation, which was effectively blocked by preincubation with 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), a classic antioxidant. Further experiments demonstrated that ROS serve as a key upstream inducer of endoplasmic reticulum (ER) stress and mitochondria-dependent apoptotic processes in AgNP/MeHg-induced injury of mouse embryo implantation and pre- and postimplantation development. Our results collectively indicate that AgNP and MeHg at non-embryotoxic concentrations can synergistically evoke ROS, ultimately causing embryotoxicity through promotion of ER stress and mitochondria-dependent apoptotic signaling cascades.

Recent advances in biological detection with rolling circle amplification: design strategy, biosensing mechanism, and practical applications.

Rolling circle amplification (RCA) is a simple and isothermal DNA amplification technique that is used to generate thousands of repeating DNA sequences using circular templates under the catalysis of DNA polymerase. Compared to alternating temperature nucleic acid amplification such as polymerase chain reaction (PCR) amplification, RCA is more suitable for on-spot detection without the need for an expensive thermal cycler. In this study, the principle and classification of RCA are introduced, and the applications of RCA in the detection of pathogenic bacteria, nucleic acid tumor markers, viruses, and proteins are reviewed. Finally, the perspectives of RCA in biological detection are anticipated. The RCA method has a great potential for biological detection. This review aims to provide references for the further development and application of the RCA technique in biosensors.

Integration of a capacitor to a 3-D DNA walker and a biofuel cell-based self-powered system for ultrasensitive bioassays of microRNAs.

A self-powered microRNA biosensor with triple signal amplification systems was assembled through the integration of three-dimensional DNA walkers, enzymatic biofuel cells and a capacitor. The DNA walker is designed from an enzyme-free target triggered catalytic hairpin assembly of modified gold nanoparticles. When triggered by the target microRNA, the DNA walker will move along the catalytic hairpin track, resulting in a payload release of glucose oxidase. The enzymatic biofuel cell contains the glucose oxidase bioanode and a bilirubin oxidase biocathode that bring a dramatic open circuit voltage to realize the self-powered bioassays of microRNA. A capacitor is further coupled with the enzymatic biofuel cell to further amplify the electrochemical signal, and the sensitivity increases 28.82 times through optimizing the matching capacitor. Based on this design, the present biosensor shows high performance, especially for detection limit and sensitivity. Furthermore, the present biosensor was successfully applied for serum samples, directly demonstrating its good application in clinical biomedicine and disease diagnosis.

Enniatin B induces dosage-related apoptosis or necrosis in mouse blastocysts leading to deleterious effects on embryo development.

The current study has focused on the effects of enniatin B (ENN B, a major mycotoxin produced by Fusarium fungi) on early embryonic development. In in vitro analysis, mouse blastocysts were incubated in medium with ENN B (0-40 µM) or 0.5% DMSO (control group) for 24 hours. In an animal study, blastocysts were collected from mice which were intravenously injected with ENN B (1, 3, 5, and 7mg/kg body weight/day) for 4 days in order to analyze apoptosis and necrosis via Annexin V/PI staining assay; and proliferation using dual differential staining. Exposure to low ENN B concentration (10 µM in vitro and 3 mg/kg/day in vivo) promoted Reactive Oxygen Species (ROS) generation and apoptosis in the Inner Cell Mass (ICM), the mass of cells inside the blastocyst, impairing post-implantation development alone. On the other hand, exposure to a higher ENN B concentration (40 µM in vitro and 7 mg/kg/day in vivo) induced ROS generation and decreased in intracellular ATP which encouraged necrotic processes in both trophectoderm (TE) and ICM of blastocysts leading to impaired implantation and post-implantation development. Moreover, 5 and 7 mg/kg/day ENN B intraperitoneal injection to female mice for 4 days has caused downregulation of CXCL1, IL-1ß and IL-8 expressions and increased ROS generation in the liver of newborn mice. Over all, ENN B can induce apoptosis and/or necrosis depending on the treatment dosage in mouse blastocysts. ENN B-induced necrosis in blastocysts may exert long-term harmful effects on next-generation newborns.

Non-embryotoxic dosage of alternariol aggravates ochratoxin A-triggered deleterious effects on embryonic development through ROS-dependent apoptotic processes.

Alternariol (AOH) and ochratoxin A (OTA), two mycotoxins found in many foods worldwide, exhibit cytotoxicity and embryotoxicity, triggering apoptosis and cell cycle arrest in several mammalian cells and mouse embryos. The absorption rate of AOH from dietary foodstuff is low, meaning that the amount of AOH obtained from the diet rarely approaches the cytotoxic threshold. Thus, the potential harm of dietary consumption of AOH is generally neglected. However, previous findings from our group and others led us to question whether a low dosage of AOH could aggravate the cytotoxicity of other mycotoxins. In the present study, we examined how low dosages of AOH affected OTA-triggered apoptosis and embryotoxicity and investigated the underlying regulatory mechanism in mouse blastocysts. Our results revealed that non-cytotoxic concentrations of AOH (1 and 2 µM) could enhance OTA (8 µM)-triggered apoptotic processes and embryotoxicity in mouse blastocysts. We also found that AOH can enhance OTA-evoked intracellular reactive oxygen species (ROS) generation and that this could be prevented by pretreatment with the potent ROS scavenger, N-acetylcysteine. Finally, we observed that this ROS generation acts as a key inducer of caspase-dependent apoptotic processes and subsequent impairments of embryo implantation and pre- and post-implantation embryonic development. In sum, our results show that non-cytotoxic dosages of AOH can aggravate OTA-triggered apoptosis and embryotoxicity through ROS- and caspase-dependent signaling pathways.

Construction of an Integrated Device of a Self-Powered Biosensor and Matching Capacitor Based on Graphdiyne and Multiple Signal Amplification: Ultrasensitive Method for MicroRNA Detection.

The detection of microRNA (miRNA) in human serum has great significance for cancer prevention. Herein, a novel self-powered biosensing platform is developed, which effectively integrates an enzymatic biofuel cell (EBFC)-based self-powered biosensor with a matching capacitor for miRNA detection. A catalytic hairpin assembly and hybrid chain reaction are used to improve the analytical performance of EBFC. Furthermore, the matching capacitor is selected as an auxiliary signal amplifying device, and graphdiyne is applied as substrate material for EBFC. The results confirm that the developed method obviously increases the output current of EBFC, and the sensitivity can reach 2.75 µA/pM, which is 786% of pure EBFC. MiRNA can be detected in an expanded linear range of 0.1-100000 fM with a detection limit of 0.034 fM (S/N = 3). It can offer a selective and sensitive platform for nucleotide sequence detection with great potential in clinical diagnostics.

Alternariol exerts embryotoxic and immunotoxic effects on mouse blastocysts through ROS-mediated apoptotic processes.

Alternariol (AOH), a mycotoxin belonging to the genus Alternaria, has been shown to induce cytotoxicity, including apoptosis and cell cycle arrest, in several mammalian cell types. However, its effects on early-stage embryonic development require further investigation. Here, we have shown that AOH exerts embryotoxic effects on mouse blastocyst-stage embryos and long-term adverse effects on immunity in one-day-old newborn mice of the next generation. Significant apoptosis and decrease in total cell number, predominantly through loss of inner cell mass (ICM), and to a minor extent, trophectoderm (TE) cells, were observed in AOH-treated blastocysts. Moreover, AOH exerted detrimental effects on pre- and post-implantation embryo development potential and induced a decrease in fetal weight in in vitro development and embryo transfer assays. Injection of pregnant mice with AOH (1, 3 and 5 mg/kg body weight/day) for 4 days resulted in apoptosis of blastocyst-stage embryos and injurious effects on embryonic development from the zygote to blastocyst stage or embryo degradation and a further decrease in fetal weight. Furthermore, AOH exerted a long-term impact on the next generation, triggering a significant increase in total oxidative stress content and expression of genes encoding antioxidant proteins. Lower expression of CXCL1, IL-1ß and IL-8 related to innate immunity was detected in liver tissue extracts obtained from one-day-old newborns of AOH-injected pregnant mice (5 mg/kg body weight/day) relative to their non-treated counterparts. In addition, ROS served as an upstream regulator of AOH-triggered apoptotic processes and impairment of embryonic development. Our collective results highlight the potential of AOH as an embryotoxic and immunotoxic risk factor during embryo and infant development stages in mice.