Pollution by perfluorooctanoic acid (PFOA), a highly persistent and toxic substance, is commonly addressed with several approaches, but the need for post-treatment affects their environmental and economic sustainability. Biodegradation, which can efficiently degrade micropollutants in water under optimal conditions, offers a promising solution, but faces challenges from the persistence of the C-F bond and the high reactivity of F-. Despite recent use of various microorganisms for PFOA degradation, performance issues persist. In this study, we aimed to simulate PFOA degradation while mixed culture anammox granules oxidize NH4+ and to confirm the effects of electric potential and Fe(Ⅲ) applications on PFOA degradation. A control reactor containing only anammox granules exhibited NH4+ and PFOA removal efficiencies of 11.52% and 19.2%, respectively, with traceable biodegradation at 3.7%. In contrast, a reactor that applied both electric potential and Fe(Ⅲ) demonstrated enhanced removal efficiencies of 41.98% and 50.93%, with traceable biodegradation at 35.03%. Bio-electrochemical properties also improved, showing an average current generation of 331.95 μA and Coulombic efficiency of 141.79 ± 7.6%. This study shows that mixed-culture anammox granules with electric potential and Fe(Ⅲ) can be introduced for efficient PFOA biodegradation and provides the basis for further studies on overcoming the limitations of PFOA biodegradation.

IMPORTANCE:In veterinary forensic science, accurately determining the postmortem interval (PMI) is crucial for identifying the causes of animal deaths. Autolysis, a significant postmortem process, influences PMI estimation, but its relationship with humidity is not well understood.OBJECTIVE:This study aimed to improve the accuracy of PMI estimates in veterinary forensic cases by looking into how different humidity levels affect autolysis in different organs of rats.METHODS:The study involved 38 male rats, examining histopathological changes in their heart, liver, and pancreas. These organs were subjected to controlled humidity levels (20%, 55%, and 80%) at a constant 22°C. Tissue samples were collected at several intervals (0 h, 12 h, 24 h, 3 days, and 8 days) for comprehensive analysis.RESULTS:Distinct autolytic characteristics in animal organs emerged under varying humidity conditions. The low-humidity environment rapidly activated autolysis more than the high-humidity environment. In addition, it was found that lower humidity caused nuclear pyknosis, cytoplasmic disintegration, and myofiber interruption. The liver, in particular, showed portal triad aggregation and hepatocyte individuation. The pancreas experienced cell fragmentation and an enlarged intracellular space. High humidity also caused the loss of striations in cardiac tissues, and the liver showed vacuolation. Under these conditions, the pancreas changed eosinophilic secretory granules.CONCLUSIONS AND RELEVANCE:The study successfully established a clear connection between the autolytic process in PMIs and relative humidity. These findings are significant for developing a more accurate and predictable method for PMI estimation in the field of veterinary forensic science.

Importance: In veterinary forensic science, accurately determining the postmortem interval (PMI) is crucial for identifying the causes of animal deaths. Autolysis, a significant postmortem process, influences PMI estimation, but its relationship with humidity is not well understood. Objective: This study aimed to improve the accuracy of PMI estimates in veterinary forensic cases by looking into how different humidity levels affect autolysis in different organs of rats. Methods: The study involved 38 male rats, examining histopathological changes in their heart, liver, and pancreas. These organs were subjected to controlled humidity levels (20%, 55%, and 80%) at a constant 22 degrees C. Tissue samples were collected at several intervals (0 h, 12 h, 24 h, 3 days, and 8 days) for comprehensive analysis. Results: Distinct autolytic characteristics in animal organs emerged under varying humidity conditions. The low -humidity environment rapidly activated autolysis more than the high -humidity environment. In addition, it was found that lower humidity caused nuclear pyknosis, cytoplasmic disintegration, and myofiber interruption. The liver, in particular, showed portal triad aggregation and hepatocyte individuation. The pancreas experienced cell fragmentation and an enlarged intracellular space. High humidity also caused the loss of striations in cardiac tissues, and the liver showed vacuolation. Under these conditions, the pancreas changed eosinophilic secretory granules. Conclusions and Relevance: The study successfully established a clear connection between the autolytic process in PMIs and relative humidity. These findings are significant for developing a more accurate and predictable method for PMI estimation in the field of veterinary forensic science.

BACKGROUND:Zinc (Zn) is an essential cofactor for physiological homeostasis in the body. Zn oxide (ZnO), an inorganic compound that supplies Zn, exists in various sizes, and its bioavailability may vary depending on the size in vivo. However, comparative studies on the nutritional effects of micro-sized ZnO (M-ZnO) and nano-sized ZnO (N-ZnO) supplementation on Zn deficiency (ZnD) animal models have not been reported.OBJECTIVES:This study investigated the nutritional bioavailability of N-ZnO and M-ZnO particles in dietary-induced ZnD mice.METHODS:Animals were divided into six experimental groups: normal group, ZnD control group, and four ZnO treatment groups (Nano-Low, Nano-High, Micro-Low, and Micro-High). After ZnD induction, N-ZnO or M-ZnO was administered orally every day for 4 weeks.RESULTS:ZnD-associated clinical signs almost disappeared 7 days after N-ZnO or M-ZnO administration. Serum Zn concentrations were higher in the Nano-High group than in the ZnD and M-ZnO groups on day 7 of ZnO treatment. In the liver and testis, Nano-Low and Nano-High groups showed significantly higher Zn concentrations than the other groups after 14-day treatment. ZnO supplementation increased Mt-1 mRNA expression in the liver and testis and Mt-2 mRNA expression in the liver. Based on hematoxylin-and-eosin staining results, N-ZnO supplementation alleviated histological damage induced by ZnD in the testis and liver.CONCLUSIONS:This study suggested that N-ZnO can be utilized faster than M-ZnO for nutritional restoration at the early stage of ZnD condition and presented Mt-1 as an indicator of Zn status in the serum, liver, and testis.

Background Previous studies have presented evidence to support the significant association between red meat intake and colon cancer, suggesting that heme iron plays a key role in colon carcinogenesis. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, exhibits anti-oxidative and anti-cancer effects. However, the effect of EGCG on red meat-associated colon carcinogenesis is not well understood. Objectives We aimed to investigate the regulatory effects of hemin and EGCG on colon carcinogenesis and the underlying mechanism of action. Methods Hemin and EGCG were treated in Caco2 cells to perform the water-soluble tetrazolium salt-1 assay, lactate dehydrogenase release assay, reactive oxygen species (ROS) detection assay, real-time quantitative polymerase chain reaction and western blot. We investigated the regulatory effects of hemin and EGCG on an azoxymethane (AOM) and dextran sodium sulfate (DSS)-induced colon carcinogenesis mouse model. Results In Caco2 cells, hemin increased cell proliferation and the expression of cell cycle regulatory proteins, and ROS levels. EGCG suppressed hemin-induced cell proliferation and cell cycle regulatory protein expression as well as mitochondrial ROS accumulation. Hemin increased nuclear factor erythroid-2-related factor 2 (Nrf2) expression, but decreased Keap1 expression. EGCG enhanced hemin-induced Nrf2 and antioxidant gene expression. Nrf2 inhibitor reversed EGCG reduced cell proliferation and cell cycle regulatory protein expression. In AOM/DSS mice, hemin treatment induced hyperplastic changes in colon tissues, inhibited by EGCG supplementation. EGCG reduced the hemin-induced numbers of total aberrant crypts and malondialdehyde concentration in the AOM/DSS model. Conclusions We demonstrated that EGCG reduced hemin-induced proliferation and colon carcinogenesis through Nrf2-inhibited mitochondrial ROS accumulation.

Asb2, ankyrin repeat, and SOCS box protein 2 form an E3 ubiquitin ligase complex. Asb2 ubiquitin ligase activity drives the degradation of filamins, which have essential functions in humans. The placenta is a temporary organ that forms during pregnancy, and normal placentation is important for survival and growth of the fetus. Recent studies have shown that approximately 25-30% of knockout (KO) mice have non-viable offspring, and 68% of knockout lines exhibit placental dysmorphologies. There are very few studies on Asb2, with insufficient research on its role in placental development. Therefore, we generated Asb2 knockout mice and undertook to investigate Asb2 expression during organogenesis, and to identify its role in early embryonic and placental development. The external morphology of KO embryos revealed abnormal phenotypes including growth retardation, pericardial effusion, pale color, and especially heart beat defect from E 9.5. Furthermore, Asb2 expression was observed in the heart from E 9.5, indicating that it is specifically expressed during early heart formation, resulting in embryonic lethality. Histological analysis of E 10.5 KO heart showed malformations such as failure of chamber formation, reduction in trabeculated myocardium length, absence of mesenchymal cells, and destruction of myocardium wall. Moreover, the histological results of Asb2-deficient placenta showed abnormal phenotypes including small labyrinth and reduced vascular complexity, indicating that failure to establish mature circulatory pattern affects the embryonic development and results in early mortality. Collectively, our results demonstrate that Asb2 knockout mice have placental defects, that subsequently result in failure to form a normal cardiac septum, and thereby result in embryo mortality in utero at around E 9.5.

The content of harmful materials was investigated for coffee beans sold in Daejeon. Total 79 samples were analysed and 213 residual pesticides and 2 heavy metals were analysed simultaneously by GC-MS/MS, GC-NPD, GC-ECD, LC-MS/MS and ICP-MS. The instrumental method was validated with limit of detection (LOD), limit of quantification (LOQ), the linearity of standard curves. LOD of the residual pesticides was between 0.0003 to 0.0021 mg/kg, LOQ of the residual pesticides was between 0.0008 to 0.0064 mg/kg. LOD of the heavy metals was between 0.0077 to 0.0079 μg/kg, LOQ of the heavy metals was between 0.0233 to 0.0239 μg/kg. The linearity correlation coefficient for the calibration curve was between 0.9929 to 0.9999 and the recovery rate was between 95.4% to 106.1%. According to the monitoring of residual pesticides and heavy metals, no pesticide was detected in all coffee bean samples. 88.6% (70 samples) of analysed total 79 coffee beans contained at least 1 heavy metal but there was no sample which exceeded the maximum residual limit. Risk assessment was also carried out based on the content of heavy metals detected in coffee beans. The carcinogenic risk assessment to heavy metals showed that all cancer-risk (CR) values were below 10–6 and it meant that the CR due to heavy metals intake was evaluated as safe. The non-carcinogenic risk assessment to heavy metals showed that all hazard index (HI) were below 1, which was considered acceptable at the current level of exposure. The %PTWI values of lead and cadmium for 55 roasted coffee bean samples were 0.09% and 0.04% respectively, compared with the reference values. This results indicate that there is almost no health risk from heavy metal intake through the consumption of coffee beans in circulation in Daejeon.

Colon cancer has been considered a leading cause of cancer-associated death. Folic acid is a vitamin necessary for cellular physiological functions and cell viability. However, the association between folic acid intake and colon cancer has been examined in several prospective cohort studies are controversial. This study investigated the effects of folate intake on colon carcinogenesis and oxidative stress in an azoxymethane (AOM)/dextran sodium sulfate (DSS) institute for cancer research (ICR) mouse model. Thirty male ICR mice (5 weeks old) were divided into the control group and the experimental group supplied 0.03% folic acid via drinking water (50 mL/week/mouse) for 6 weeks. To induce colonic pre-neoplastic lesions, the animals were subcutaneously injected three times weekly with AOM (10 mg/kg body weight), followed by 2% DSS in drinking water for a week. Folic acid supplementation significantly suppressed the total number of aberrant crypt foci and aberrant crypts. Histological image data showed that folic acid supplementation attenuated neoplastic change. In addition, we measured the thiobarbituric acid reactive substances concentration of dry feces samples to identify the effect of folic acid on reactive oxygen accumulation. The folic acid supplementation group had reduced reactive oxygen species levels in dry feces compared to the control group. In conclusion, these findings indicate that folic acid suppresses colon carcinogenesis and oxidative stress in an AOM/DSS mouse model.

Colorectal cancer causes the most cancer-associated death worldwide, having a high cancer incidence. Pectin is a complex polysaccharide present in various fruits, emerging as an anti-carcinogenic candidate. Although pectin has a suppressive capacity for colon carcinogenesis, the effect of reactive oxygen species (ROS) generation and colonic aberrant foci formation in the colon carcinogenesis mouse model remains unclear. Therefore, this study investigates the regulatory effect of pectin supplementation on colon carcinogenesis induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in mice. In an animal experiment, thirty male institute for cancer research (ICR) mice were divided into two experimental groups; AOM/DSS (control group) and AOM/DSS + pectin (5% in drinking water). Furthermore, the number of aberrant crypt foci (ACF) and aberrant crypt (AC) on colonic mucosa were counted, and thiobarbituric acid-reactive substances (TBARS) assay was performed to estimate lipid peroxidation in feces. Pectin treatment significantly decreased the number of ACF and AC per colon compared with the control. Additionally, fecal TBARS level in the pectin group was significantly lower than those in the control group. Conclusively, these findings indicate that pectin-inhibited hyperplastic alteration and oxidative stress suppress colitis-associated colon carcinogenesis.

Feline parvovirus (FPV) is a small, non-enveloped, single-stranded DNA virus that infects cats. We recently isolated a feline parvovirus Fe–P2 strain from a dead stray cat in Iksan, 2017. Its partial genomic sequence (4,643 bases) was obtained, and phylogenetic analysis based on the VP2 nucleotide sequence showed that the FPV Fe-P2 strain was closely related to the FPV isolate Gigucheon in cat, 2017 (MN400978). In addition, we performed a serum neutralization (SN) test with the FPV isolates in various mammalian sera. These were from raccoon dog, water deer, Eurasian otter, Korean hare, leopard cat, and Asian badger, which were kindly provided by Chungnam Wild Animal Rescue Center. Notably, serological evidence of its infection was found in Asian badger, Meles leucurus (2/2) and leopard cat, Prionailurus bengalensis (5/8) through SN tests, whereas there was no evidence in raccoon dog, water deer, Eurasian otter, and Korean hare based on the collected sera in this study. These findings might provide partial evidence for the possible circulation of FPV or its related viruses among wild leopard cat and Asian badger in Korea. There should be additional study to confirm this through direct detection of FPVs in the related animal samples.

Expanding on previous demonstrations of the therapeutic effects of adeno‐associated virus (AAV) carrying small‐hairpin RNA (shRNA) in downregulating the mechanistic target of rapamycin (mTOR) in in vivo retinal vascular disorders, vascular endothelial growth factor (VEGF)‐stimulated endothelial cells were treated with AAV2‐shmTOR to examine the role of mTOR inhibition in retinal angiogenesis. AAV2‐shmTOR exposure significantly reduced mTOR expression in human umbilical vein endothelial cells (HUVECs) and decreased downstream signaling cascades of mTOR complex 1 (mTORC1) and mTORC2 under VEGF treatment. Moreover, the angiogenic potential of VEGF was significantly inhibited by AAV2‐shmTOR, which preserved endothelial integrity by maintaining tight junctions between HUVECs. These data thus support previous in vivo studies and provide evidence that AAV2‐shmTOR induces therapeutic effects by inhibiting the neovascularization of endothelial cells.

Various microorganisms are involved in nitrogen removal, and their group compositions depend closely on operating parameters. The structures and functions of nitrification microorganisms in full-scale anaerobic-anoxic-oxic (A2/O) and oxidation ditch processes were analyzed using metagenomics and canonical correspondence analysis. The community structure of ammonia-oxidizing archaea in the oxidation ditch was 3.8 (winter) - 6.3 (summer) times higher than in A2/O, and the complete ammonia oxidizer was only found in the oxidation ditch process. The canonical correspondence analysis of various environmental variables showed that Nitrosomonadales, Crenarchaeota, and Nitrospira inopinata correlate highly with nitrification, and Nitrospira was involved in NO2--N oxidation rather than Nitrobacter. The longer solid and hydraulic retention times in the oxidation ditch were more effective in achieving a wider range of novel nitrification than A2/O. This result indicates that microbial communities of novel nitrifiers and ammonia-oxidizing archaea improved in the oxidation ditch process, significantly contributing to stable nitrogen removal.

Colon cancer is known as the third most widespread cancer in the world.The interaction of heme-iron and ascorbic acid (AA) in colon carcinogenesis is not evident.Hemin (ferric chloride heme) is an iron-containing porphyrin with chlorine that can be formed from a heme group.The purpose of this study was to investigate the protective effect of AA on the formation of pre-neoplastic lesions induced by azoxymethane (AOM)/dextran sodium sulfate (DSS) plus hemin in mice.Forty-five ICR male mice were divided into three experimental groups; AOM/ DSS treatment (control group), hemin (2 g hemin/kg of b.w.), hemin + AA (1.0% in drinking water).The mice had three s.c.injections (0-2nd weeks of the experiment) of AOM (10 mg/kg b.w.) weekly and 2% DSS as drinking water for the next one week and the animals fed on AIN-76A purified rodent diet for 6 weeks.The numbers of aberrant crypt foci (ACF) and aberrant crypts (ACs) in colonic mucosa were counted after methylene blue staining.Lipid peroxidation in feces was measured by the thiobarbituric acid-reactive substances (TBARS) assay.The numbers of ACF and ACs per colon significantly increased in Hemin group compared to the control group.However, the numbers of ACF and ACs per colon notably decreased in hemin + AA group compared to the control group or hemin group (p<0.05).In feces, the TBARS value of hemin group was higher than the control group (p<0.01).The TBARS value of hemin + AA group was slightly decreased compared to Hemin group.These results indicate that hemin can promote the experimental colon carcinogenesis in ICR mice.On the other hand, additional supplement of AA via drinking water has a protective effect against the colon carcinogenesis.The related mechanisms need to be illustrated by further studies in future.

Globally, colon cancer is increased gradually and known as one of the major causes of cancer death. Stevia, a substitute of sugar, is known to have many components including alpha-tocopherol and anthocyanin etc, as antioxidants. This study's purpose is to investigate whether stevia plant extract can have a protective effect against colon carcinogenesis induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in mice. Total 30 male ICR mice were divided into 2 groups; AOM/DSS treatment (control group), AOM/DSS + stevia extract (0.5%, in drinking water). After acclimation for 1 week, five weeks old mice received three intraperitoneal AOM (10 mg/kg b.w.) injections weekly for 3 weeks (0–2nd weeks of the experiment) and 2% DSS as drinking water for the next one week. AIN-76A purified rodent diet and 0.5% stevia extract water were supplied to the animals for 6 weeks. The colons of mice were collected and the number of aberrant crypt foci (ACF) and aberrant crypts (ACs) in colonic mucosa were counted after staining with methylene blue. Malondialdehyde (MDA) concentration in feces were determined. The numbers of ACF and ACs were significantly (p<0.01) decreased in stevia-treated group compared with the control group. The MDA concentration in feces was also significantly (p<0.01) decreased in stevia-treated group compared with the control group. In histopathology of colonic epithelium, hyperplasia of colonic epithelium was less observed in steviatreated group. These results indicate that stevia has a protective effect against colon carcinogenesis induced by AOM/DSS in mice and further study needs to illustrate the protective mechanisms.

Biochemical methane potential tests and lab-scale continuous experiments were conducted to improve the yield and energy efficiency of anaerobic digestion through thermal hydrolysis pre-treatment. Methane generation, yield, and solubilization efficiency were evaluated through lab-scale tests. The pre-treated samples presented 50% biodegradability at 140 °C and 61.5% biodegradability at 165 °C. The increase in biodegradability was insignificant at 165 °C or higher temperature, and it was confirmed that the optimum conditions were achieved at 165 °C and 20 min of solubilization. The lab-scale continuous experiments confirmed that polymers were decomposed into low-molecular-weight compounds due to thermal hydrolysis, and pH decreased. NH4HCO3 produced by thermal hydrolysis acted as an alkali to enable a more stable operation compared to that before thermal hydrolysis. Total chemical oxygen demand as chromium (TCODCr) and soluble chemical oxygen demand as chromium (SCODCr) indicated 35.4% and 23.1% removal efficiency in terms of organic matter removal, respectively. Methane yield was approximately 0.35 kg m−3 at 2.0–4.0 kg (m3 d)−1 and 0.26 kg m−3 at 5.0 kg (m3 d)−1. The solubilization rate of 40.9% by thermal hydrolysis was confirmed through the lab-scale tests to determine its full-scale applicability.

Anaerobic digestion (AD) is one of the most widely adopted bioenergy recovery technologies globally. Despite the wide adoption, AD has been challenged by the unstable performances caused by imbalanced substrate and/or electron availability among different reaction steps. Bioelectrochemical anaerobic digestion (BEAD) is a promising concept that has demonstrated potential for balancing the electron transfer rates and enhancing the methane yield in AD during shocks. While great progress has been made, a wide range of, and sometimes inconsistent engineering and technical strategies were attempted to improve BEAD. To consolidate past efforts and guide future development, a comprehensive review of the fundamental bioprocesses in BEAD is provided herein, followed by a critical evaluation of the engineering and technical optimizations attempted thus far. Further, a few novel directions and strategies that can enhance the performance and practicality of BEAD are proposed for future research to consider. This review and outlook aim to provide a fundamental understanding of BEAD and inspire new research ideas in AD and BEAD in a mechanism-informed fashion.

Identification of a new agent from natural products for the protection of embryonic anomalies is potentially valuable. To investigate the protective effect exerted by lycopene against nicotine-induced malformations, mouse embryos in embryonic day 8.5 with yolk sac placentas were cocultured with 1 mM nicotine and/or lycopene (1×10−6,1×10−5 μM) for 48 h. The morphological defects and apoptotic cell deaths in the embryo and yolk sac placenta of the nicotine group were significantly increased. Exposure to nicotine resulted in reduced superoxide dismutase (SOD) activity and cytoplasmic SOD and cytoplasmic glutathione peroxidase mRNA levels, but increased lipid peroxidation level in embryos. Moreover, treatment with nicotine resulted in aggravated expressions of the mRNA or protein level of antiapoptotic (BCL2-associated X protein, B-cell lymphoma-extralarge, and caspase 3), anti-inflammatory (nuclear factor kappa-light-chain-enhancer of activated B cells and tumor necrosis factor-alpha), and vasculogenic (vascular endothelial growth factor-alpha, insulin-like growth factor-1, alpha smooth muscle actin, transforming growth factor-beta 1, and hypoxia inducible factor-1 alpha) factors in the embryo and yolk sac placenta. However, all the parameters were significantly improved by treatment with lycopene, as compared to the nicotine group. These findings indicate the potential of lycopene as a protective agent against embryonic anomalies and yolk sac vasculogenic and placenta-forming defects induced by nicotine through modulations of oxidative, apoptotic, vasculogenic, and inflammatory activities.

Endocrine-disrupting chemicals (EDC) are substances similar to steroid hormones that can disturb normal physiological functions of male and female reproductive organs. Endocrine-disrupting chemicals tend to bind to steroid hormone receptors. Sex steroid hormones modulate calcium signalling in the cardiac muscle in early embryo development. Among the steroid hormones, progesterone (P4) has been reported to affect blood pressure and other aspects of the cardiovascular system. The EDC that have similar structure to P4, such as octyl-phenol (OP) and bisphenol A (BPA), are potentially harmful to development of the heart. To confirm the effect of OP and BPA on early differentiation of mouse embryonic stem cells (mES) into cardiomyocytes, the hanging-drop method with mESC cell line (ES-E14TG2a) was used for forming embryoid bodies. Pluripotent mESC were cultured in basal medium with leukemia inhibitory factor and grown on mitomycin C-treated mouse embryonic fibroblasts in a 60-mm plate at 37°C in a 5% CO2 humidified tissue culture incubator with basal medium consisting of DMEM/F-12 supplemented with nonessential amino acids, 10% heat-inactivated and certified fetal bovine serum (FBS), 2-mercaptoethanol, penicillin, and streptomycin. The mouse embryoid bodies (mEB) were suspended onto 6-well plates and cultured with differentiation medium containing steroid-free FBS without leukemia inhibitory factor. Progesterone, OP, and BPA were added on Day 2 from mEB attachment. In addition, mifepristone (RU486), an antagonist for progesterone receptor (PR), was used to confirm the effect of P4 through PR. To determine whether RU486 is capable of attenuating the inhibition effect of P4, RU486 was added for 1 day starting on Day 11. Assessment of cardiomyocyte differentiation was determined by checking the number of beating cell populations divided by the total number of attached mEB. Total RNA was extracted using Trizol reagent and synthesised to cDNA using Moloney murine leukemia virus (MMLV) reverse transcriptase. Messenger RNA level was assessed using quantitative real-time PCR. To investigate calcium signalling, the mRNA levels of calcium channel gene Trpv2 and contraction-related genes Ryr2, Cam2, and Mylk3 were analysed. Beating ratio was decreased in P4, OP, and BPA treatments. Data were analysed by one-way ANOVA followed by Tukey’s multiple comparison tests, and P<0.05 was considered statistically significant in least 3 different replicates. The mRNA level of Pgr was significantly increased in P4, OP, and BPA-treated group. However, the mRNA level of calcium channel gene Trpv2 was significantly decreased in the P4, OP, and BPA-treated group. Expression of contraction-related genes such as Ryr2, Cam2, and Mlck3 were significantly decreased in the P4, OP, and BPA-treated group. In addition, treatment with RU486 rescues altered calcium channel gene and contraction-related genes. Taken together, these results suggest that OP and BPA may affect the differentiation of mESC into cardiomyocyte and disrupts differentiation of cardiomyocytes.

Miscarriage due to blastocyst implantation failure occurs in up to two-thirds of all miscarriage cases in humans. Calcium (Ca2+) has been shown to involve many cellular signal transduction pathways as well as regulation of cell adhesion, which is necessary for the physiology process of endometrial epithelial cell transformation and stromal cell decidualization during embryo implantation. Exposure to endocrine-disrupting chemicals (ED) can regulate the expression of genes associated with calcium transport in during pregnancy such as TRPV5, TRPV6, PMCA, and NCX1. Additionally, exposure to ED during early gestation results in disrupted intrauterine implantation, uterine receptive, leading to implantation failure. In this study, oestrogen (E2), bisphenol A (BPA), octylphenol (OP), and/or ICI 182,780 (oestrogen receptor antagonist, ICI) were injected subcutaneously from gestation Day 1 to gestation Day 3 post coitus. The number of implantation sites was significantly lower in the OP group, and no implantation site was observed in the E2 and ED+ICI groups. There were differences in the expression of calcium transient transport channel between maternal uterine and implantation. The level of TRPV6 and TRPV5 mRNA and protein was significantly increased by ED and/or ICI treatment in the uterus. The levels of TRPV5 and TRPV6 gene expression were significantly increased by ED with/without ICI treatment in the uterus. However, TRPV5 and TRPV6 gene expression was significantly lower in implantation site samples. The NCX1 and PMCA1 mRNA levels were significantly decreased by OP and BPA in the implantation site samples. Both mRNA and protein levels of MUC1 were markedly higher in all groups, except the BPA group when compared with the vehicle group in the uterus. The LIF and HOXA-10 mRNA were significantly low in E2; BPA+ICI; OP; and/or ICI in both the uterus and implantation site. Expression of the oestrogen receptor (ERa) and progesterone receptor (PR) was significantly lower in all groups except the BPA group when compared with the vehicle group. Taken together, E2, BPA, and OP disrupt the success of implantation through altered expression of calcium transport genes.