Low doses of BPF-induced hypertrophy in cardiomyocytes derived from human embryonic stem cells via disrupting the mitochondrial fission upon the interaction between ERß and calcineurin A-DRP1 signaling pathway.

Bisphenol F (BPF) is a replacement to bisphenol A, which has been extensively used in industrial manufacturing. Its wide detection in various human samples raises increasing concern on its safety. Currently, whether a low dose of BPF compromises cardiac function is still unknown. This study provides the first evidence that low-dose BPF can induce cardiac hypertrophy by using cardiomyocytes derived from human embryonic stem cells (hES). Non-cytotoxic BPF increased cytosolic Ca 2+ influx ([Ca2+ ]c), which was most remarkable at low dose (7 ng/ml) rather than at higher doses. Significant changes in the morphological parameters of mitochondria and significant decreases in ATP production were induced by 7 ng/ml BPF, representing a classic hypertrophic cardiomyocyte. After eliminating the direct effects on mitochondrial fission-related DRP1 by administration of the DRP1 inhibitor Mdivi-1, we examined the changes in [Ca 2+ ]c levels induced by BPF, which enhanced the calcineurin (Cn) activity and induced the abnormal mitochondrial fission via the CnAß-DRP1 signaling pathway. BPF triggered excessive Ca 2+ influx by disrupting the L-type Ca 2+channel in cardiomyocytes. The interaction between ERß and CnAß cooperatively involved in the BPF-induced Ca 2+ influx, which resulted in the abnormal mitochondrial fission and compromised the cardiac function. Our findings provide a feasible molecular mechanism for explaining low-dose BPF-induced cardiac hypertrophy in vitro, preliminarily suggesting that BPF may not be as safe as assumed in humans.

Low doses of BPA induced abnormal mitochondrial fission and hypertrophy in human embryonic stem cell-derived cardiomyocytes via the calcineurin-DRP1 signaling pathway: A comparison between XX and XY cardiomyocytes.

Humans are inevitably exposed to bisphenol A (BPA) via multiple exposure ways. Thus, attention should be raised to the possible adverse effects related to low doses of BPA. Epidemiological studies have outlined BPA exposure and the increased risk of cardiovascular diseases (such as cardiac hypertrophy), which has been confirmed to be sex-specific in rodent animals and present in few in vitro studies, although the molecular mechanism is still unclear. However, whether BPA at low doses equivalent to human internal exposure level could induce cardiac hypertrophy via the calcineurin-DRP1 signaling pathway by disrupting calcium homeostasis is unknown. To address this, human embryonic stem cell (H1, XY karyotype and H9, XX karyotype)-derived cardiomyocytes (CM) were purified and applied to study the low-dose effects of BPA on cardiomyocyte hypertrophy. In our study, when H1- and H9-CM were exposed to noncytotoxic BPA (8 ng/ml), markedly elevated hypertrophic-related mRNA expression levels (such as NPPA and NPPB), enhanced cellular area and reduced ATP supplementation, demonstrated the hypertrophic cardiomyocyte phenotype in vitro. The excessive fission produced by BPA was promoted by CnAß-mediated dephosphorylation of DRP1. At the molecular level, the increase in cytosolic Ca2+ levels by low doses of BPA could discriminate between H1- and H9-CM, which may suggest a potential sex-specific hypertrophic risk in cardiomyocytes in terms of abnormal mitochondrial fission and ATP production by impairing CnAß-DRP1 signaling. In CnAß-knockdown cardiomyocytes, these changes were highly presented in XX-karyotyped cells, rather than in XY-karyotyped cells.

Safety of concomitant administration of inactivated hepatitis A vaccine with other vaccines in children under 16 years old in post-marketing surveillance.

BACKGROUND: Concomitant administration refers to the receipt of two or more vaccines during a single healthcare encounter, which is an efficient way to increase vaccination coverage in children. However, the post-marketing safety studies of concomitant administration are scarce. Inactivated hepatitis A vaccine (Healive®) has been used widely in China and other countries for more than a decade. We aimed to explore the safety of Healive® co-administered with other vaccines compared to Healive® alone in children under 16 years old. METHODS: We retrieved Adverse Events Following Immunization (AEFI) cases and vaccination doses of Healive® during 2020-2021 in Shanghai, China. The AEFI cases were divided into concomitant administration group and Healive® alone group. We used administrative data on vaccine doses as denominators to calculate and compare crude reporting rates between groups. We also compared baseline gender and age distribution, clinical diagnoses, and time interval from vaccination to onset of symptoms between groups. RESULTS: A total 319,247 doses of inactivated hepatitis A vaccine (Healive®) were used and 1,020 AEFI cases (319.50 per million doses) associated with Healive® were reported during 2020-2021 in Shanghai. There were 259,346 doses concomitantly administered with other vaccines and 830 AEFI cases (320.04 per million doses) were reported. There were 59,901 doses of Healive® that vaccinated alone, with 190 AEFI cases (317.19 per million doses). There was only one case with serious AEFI in concomitant administration group, with a rate of 0.39 per million doses. Reported rates of AEFI cases were similar between groups in general (p > 0.05). CONCLUSION: Concomitant administration of inactivated hepatitis A vaccine (Healive®) with other vaccines has a similar safe profile as Healive® alone.

Establishment of a 13 genes-based molecular prediction score model to discriminate the neurotoxic potential of food relevant-chemicals.

Although many neurotoxicity prediction studies of food additives have been developed, they are applicable in a qualitative way. We aimed to develop a novel prediction score that is described quantitatively and precisely. We examined cell viability, reactive oxygen species activity, intracellular calcium and RNA transcription level of potential prediction related genes to develop a high-throughput neurotoxicity test method in vitro to screen the neurotoxicity of hazardous factors in food using AI-based machine learning. We trained artificial intelligence models (random forest and neural network) to predict neurotoxicity precisely, establishing a universal classification assessment score (CA-Score) that relies on the expression status of only 13 of prediction related genes. The CA-Score system is almost universally applicable to food risk factors (p<0.05) in a manner independent of platform (microarray or RNA sequencing) by being compared with cut-off value 23.487 to judge whether it's neurotoxic or not. We finally validated our prediction with the external validation of CA-Score on neural precursor cells derived from embryonic stem cells. Therefore, we draw a conclusion that the AI-based machine learning including neural network and random forest is likely to provide a useful tool for large-scale screening of neurotoxicity in food risk factors.

Combined effects of BPA and PFOS on fetal cardiac development: In vitro and in vivo experiments.

Analyses of the combined effects of different EDCs are both important and difficult. This study attempts to evaluate the individual and combined effects of BPA and PFOS on heart development. Sprague-Dawley rats received individual or combined PFOS and BPA for 19 days during pregnancy. The results show that the combined BPA and PFOS exposure could lead to a morphological change in the fetal rat heart. An increase in the interventricular septal thickness (IVS) of approximately 20 % (391 µm in control vs 464 µm in combined exposure) was observed in the fetal rat hearts after the combined exposure to nearly 2000 µg/L PFOS and 100 µg/L BPA through drinking water. The total collagen and dynamin-related protein 1 (Drp1) mRNA level was increased in the fetal hearts exposed to the combination of 2000 µg/L PFOS and 100 µg/L BPA. However, the cell number in the IVS did not significantly change. Based on the previous literature, we believe that the combined exposure to BPA and PFOS had a synergistic effect on the thickness of the IVS. The combined exposure to 40 µg/L PFOS and 2 µg/L BPA failed to cause significant damage to the embryonic heart. The individual and combined effects and the mechanism of the effects of BPA and PFOS on heart development were further investigated by an in vitro study. Embryonic stem cells were administered individual or combined 10 ng/mL BPA and 100 ng/mL PFOS for 14 days during the cardiac differentiation period. The results show that exposure to the combination of 100 ng/mL PFOS and 10 ng/mL BPA could increase the cardiomyocyte size and collagen content. A selective inhibitor of Drp1, Mdivi-1, could inhibit the cardiomyocyte size enlargement but not the collagen content increase caused by the combined exposure. Thus, we believe that although the combined exposure to PFOS and BPA could affect mitochondrial biogenesis and collagen expression, these two effects seem to be relatively independent. Based on these results, this research concludes that combined exposure to PFOS and BPA could specifically lead to increased collagen and IVS thickening in heart development.

Use of embryonic stem cell-derived cardiomyocytes to study cardiotoxicity of bisphenol AF via the GPER/CAM/eNOS pathway.

Bisphenol AF (BPAF) is a derivative of bisphenol A (BPA) that is widely used in fluorinated polymers, fluorinated rubber, electronic equipment, plastic optical fibers, etc. Studies have shown that BPAF exposure is associated with a number of diseases; however, little is known about the effects of BPAF on cardiomyocytes. We investigated the impact of chronic exposure to BPAF on cardiomyocytes derived from embryonic stem cells (ESCs). The present study showed that chronic exposure to various concentrations of BPAF (0, 8, 200 and 1000 ng/ml) induces cardiomyocyte hypertrophy. The ratios of microfilaments to mitochondrial length and the ratio of microfilaments to cell nuclei and MYH7b levels indicate that BPAF exposure alters the morphology of the cells and mitochondria. Furthermore, BPAF exposure at concentrations from 8 to 1000 ng/ml results in an increase in G protein-coupled estrogen receptor (GPER) expression. Additionally, our results suggest that these effects of BPAF mediate cardiomyocyte hypertrophy apparently due to an increase in the production of reactive nitrogen species (RNS) via an increase in endothelial NO synthase (eNOS). These results imply that ESC-based myocardial differentiation can be an excellent cellular model to study BPAF-induced cardiotoxicity at the cellular and molecular levels.

Low-dose exposure to triclosan disrupted osteogenic differentiation of mouse embryonic stem cells via BMP/ERK/Smad/Runx-2 signalling pathway.

Triclosan (TCS) has been used widely in personal care products for its broad-spectrum antimicrobial activity. The detection of TCS in the umbilical cord sera, amniotic fluid, and placenta, has raised concerns about the risk to foetal development. In the current study, the embryonic stem cells test (EST) were utilized primarily for the evaluation of the adverse effects of TCS on cardiogenesis and osteogenesis in vitro. TCS was predicted to be weakly embryotoxic in cardiogenesis and strongly embryotoxic in osteogenesis. The 50% inhibition value of osteogenic differentiation was 110 times lower than that of cardiac differentiation, which suggested that the development of the skeletal system was more sensitive to TCS-induced disruption. The mechanism through which TCS exerted toxicity on osteogenesis was studied further. Decreased calcification in ESC-derived osteoblasts was observed after exposure to TCS at a low dose, equal to the human internal exposure level. TCS was observed to specifically target ERK activation, rather than JNK or p38. Further, the downregulation of p-Smad-1, together with strong inhibition on Runx-2 and Bglap-2 expression, was observed via BMP/ERK/Smad signalling when cells were exposed to TCS. The change in Runx-2 induced by a low-dose TCS highlighted a specific target for exploring its adverse effect on skeletal development.