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Mito-TEMPO is a well-known mitochondria-specific superoxide scavenger. However, the effect of Mito-TEMPO on porcine embryo development, to our knowledge, has not been studied yet. In the present study, porcine embryos were classified into two groups (G1 and G2) based on the cytoplasm lipid contents at the zygote stage. The development of blastocysts derived from G2 zygotes was reduced (G2:16.2 ± 7.9% vs G1: 26.5 ± 5.9%; 1.6-fold, p < 0.05) compared to those from G1 zygotes. In G2 embryos, the proportion of TUNEL-positive cells was also higher than that of G1 embryos. Superoxide in G2 embryos was significantly increased compared to that in G1 embryos. Mitochondrial membrane potential and ATP production were lower in G2 embryos than in G1 embryos. Phosphorylation of Drp1 at Ser 616 increased in G1 embryos during the cleavage stages compared to that in the zygote but was not significantly different in G2 embryos. Then, the effects of Mito-TEMPO were investigated in G2 embryos. Blastocyst formation rate (G2: 19.1 ± 5.1% vs G2 + Mito-TEMPO: 28.8 ± 4.0%; 1.5-fold, p < 0.05) and mitochondrial aggregation were recovered after superoxide reduction by Mito-TEMPO treatment. Thus, we showed that Mito-TEMPO improves blastocyst development by superoxide reduction in porcine embryos in vitro .

Bisphenol A (BPA) is synthetic organic compound that exhibits estrogen-like properties and it induces mitochondrial superoxide production. Melatonin (Mela) protects against BPA-mediated cell damage and apoptosis. However, the antioxidative effects of Mela against BPA-induced superoxide production in porcine oocytes are still not known. In this study, we investigated the antioxidative effects of Mela against BPA-derived superoxide on oocyte maturation in pigs. To investigate the effects of the superoxide specific scavenger, Mito-TEMPO, on porcine oocyte maturation in response to BPA exposure apoptosis proteins, we treated the oocytes with Mito-TEMPO (0.1 µM) after pre-treating them with BPA (75 µM) for 22 h. As expected, the reduction in meiotic maturation and cumulus cell expansion of cumulus-oocyte-complexes (COCs) in the BPA (75 µM) treated group was recovered (p < 0.01) by treatment with Mito-TEMPO (0.1 µM). An increase in the levels of mitochondrial apoptotic proteins (AIF, cleaved Cas 3 and cleaved Parp1) in response to BPA-induced damage was also reduced by Mito-TEMPO treatment in porcine COCs. Interestingly, we confirmed the positive effects of Mela with respect to superoxide production upon BPA exposure during oocyte maturation and also confirmed the reduction in mitochondrial apoptosis in Mela (0.1 µM)-treated porcine COCs. These results provide evidence for the first time that antioxidative effects of Mela on BPA-derived superoxide improve porcine oocyte maturation.

Background Post-ovulatory aging (POA) is associated with reduced fertilization rates and poor embryo quality both in vivo and in vitro . However, the relationship between nicotinamide adenine dinucleotide (NAD + ) and the filamentous actin (F-actin) cytoskeleton in POA-induced oocytes remains unknown. Here, we investigated the mechanisms by which the NAD + salvage pathways function in poor oocyte maturation upon POA through the F-actin cytoskeleton. Methods Porcine oocytes were aged by extending in vitro maturation (IVM) for an additional 24 h to create a POA model. F-actin and adducin 1 (ADD1)-related spindle assembly were analyzed using immunofluorescence, western blotting, and RNA sequencing to identify key gene categories in the POA and IVM groups. To assess NAD + function in restoring oocyte maturation, nicotinamide mononucleotide (NMN) was added and the maturation efficiency was evaluated. Expression of spindle assembly factors, F-actin cytoskeleton factors, aging markers, and NAD + -related genes was analyzed via quantitative polymerase chain reaction, immunofluorescence, and western blotting. Results We revealed unique interactions between the F-actin/ADD1-related cytoskeleton and aging factors (clusterin ( CLU ) and FAM111 trypsin-like peptidase A ( FAM111A )) in poor-quality oocytes. POA oocytes were established with an extension of 24 h based on 44 h of IVM. They exhibited actin collapses and abnormal cortical F-actin, ADD1, and acetyl(Ac)-α-tubulin protein levels, which resulted in defective spindle assembly. RNA sequencing analysis was performed to identify differentially expressed genes involved in the oocyte viability response to aging, the cytoskeleton, and NAD metabolic processes using IVM and/or POA oocytes. This showed that NAD-binding genes were differentially expressed after POA induction, eight of which were downregulated compared with IVM oocytes. Importantly, activation of NAD + pathways upon addition of NMN to the medium at 24 h after IVM rescued the maturation capability of POA oocytes with perturbations of spindle assembly and cortical F-actin. Conclusion F-actin polymerization through NAD + generated from NMN is an essential factor in determining oocyte quality. This effect is mediated by microtubules related to spindle assembly in POA oocytes.

Mitochondrial division inhibitor 1 (Mdivi-1) reportedly provides a close connection between oocyte maturation and mitochondrial function in pigs. N -acetyl-5-methoxy-tryptamine (melatonin) is known to be a representative antioxidant with the ability to rehabilitate meiotic maturation of porcine oocytes. However, the ability of melatonin to recover Mdivi-1-mediated disruption of spindle formation during meiotic maturation of porcine oocytes during in vitro maturation (IVM) has not been studied. Here, we first investigated changes in mitochondrial length, such as fragmentation and elongation form, in mature porcine oocytes during IVM. Mature oocytes require appropriate mitochondrial fission for porcine oocyte maturation. We identified a dose-dependent reduction in meiotic maturation in porcine oocytes following Mdivi-1 treatment (50, 75, and 100 μM). We also confirmed changes in mitochondrial fission protein levels [dynamin-related protein 1 phosphorylation at serine 616 (pDRP1-Ser616) and dynamin-related protein 1 (DRP1)], mitochondrial membrane potential, and ATP production in 75 μM Mdivi-1-treated oocytes. As expected, Mdivi-1 significantly reduced mitochondrial function and DRP1 protein levels and increased spindle abnormalities in porcine oocytes. In addition, we confirmed that melatonin restores abnormal spindle assembly and reduces meiotic maturation rates by Mdivi-1 during porcine oocyte maturation. Interestingly, the expression levels of genes that reduce DNA damage and improve tubulin formation were enhanced during porcine meiotic maturation. Taken together, these results suggest that melatonin has direct beneficial effects on meiotic maturation through tubulin formation factors during porcine oocyte maturation.

While triclosan (TCS) exerts detrimental effects on female reproduction, the effect of TCS-derived toxins on porcine oocytes during in vitro maturation (IVM) is unclear. This study investigated the effects of TCS on mitochondrion-derived reactive oxygen species (ROS) production and apoptosis pathways during porcine oocyte maturation. Porcine oocytes were treated with TCS (1, 10, and 100 μM) and triphenylphosphonium chloride (Mito-TEMPO; 0.1 μM), and matured cumulus oocyte complexes (COCs) were stained with orcein, dichlorofluorescein diacetate (DCF-DA), and Mito-SOX. Proteins and mRNA levels of factors related to cumulus expansion and mitochondrion-mediated apoptosis and antioxidant enzymes were analyzed by western blotting and reverse-transcription polymerase chain reaction (RT-PCR), respectively. Meiotic maturation and cumulus cell expansion significantly decreased for COCs after TCS treatment along with an increase in mitochondrial superoxide levels at 44 h of IVM. Further, mitochondrion-related antioxidant enzymes and apoptosis markers were significantly elevated in porcine COCs following TCS-mediated oxidative damage. The protective effect of Mito-TEMPO as a specific superoxide scavenger from TCS toxin improved the maturation capacity of porcine COCs. Mito-TEMPO downregulated the mitochondrial apoptosis of TCS-exposed porcine COCs by reducing superoxide level. In conclusion, our data demonstrate that TCS mediates toxicity during porcine oocyte maturation through superoxide production and mitochondrion-mediated apoptosis.

Changes in F-actin distribution and cortical F-actin morphology are important for blastocyst developmental competence during embryogenesis. However, the effect of paclitaxel as a microtubule stabilizer on embryonic development in pigs remains unclear. We investigated the role of F-actin cytoskeleton stabilization via P38 MAPK activation using paclitaxel to improve the developmental potential of blastocysts in pigs. In this study, F-actin enrichment and adducin expression based on blastomere fragment rate and cytokinesis defects were investigated in cleaved embryos after in vitro fertilization (IVF). Adducin and adhesive junction F-actin fluorescence intensity were significantly reduced with increasing blastomere fragment rate in porcine embryos. In addition, porcine embryos were cultured with 10 and 100 nM paclitaxel for two days after IVF. Adhesive junction F-actin stabilization and p-P38 MAPK activity in embryos exposed to 10 nM paclitaxel increased significantly with blastocyst development competence. However, increased F-actin aggregation, cytokinesis defects, and over-expression of p-P38 MAPK protein by 100 nM paclitaxel exposure disrupted blastocyst development in porcine embryos. In addition, exposure to 100 nM paclitaxel increased the misaligned α-tubulin of spindle assembly and adhesive junction F-actin aggregation at the blastocyst stage, which might be caused by p-P38 protein over-expression-derived apoptosis in porcine embryos.

Conventional antiferroelectric materials with atomic-scale anti-aligned dipoles undergo a transition to a ferroelectric (FE) phase under strong electric fields. The moiré superlattice formed in the twisted stacks of van der Waals crystals exhibits polar domains alternating in moiré length with anti-aligned dipoles. In this moiré domain antiferroelectic (MDAF) arrangement, the distribution of electric dipoles is distinguished from that of two-dimensional FEs, suggesting dissimilar domain dynamics. Here we performed an operando transmission electron microscopy investigation on twisted bilayer WSe2 to observe the polar domain dynamics in real time. We find that the topological protection, provided by the domain wall network, prevents the MDAF-to-FE transition. As one decreases the twist angle, however, this transition occurs as the domain wall network disappears. Exploiting stroboscopic operando transmission electron microscopy on the FE phase, we measure a maximum domain wall velocity of 300 μm s–1. Domain wall pinnings by various disorders limit the domain wall velocity and cause Barkhausen noises in the polarization hysteresis loop. Atomic-scale analysis of the pinning disorders provides structural insight on how to improve the switching speed of van der Waals FEs.Polar domains have been observed in twist-stacked van der Waals layers, but their dynamics are unexplored. Here, using operando electron microscopy, it is found that polar domains in an antiferroelectric arrangement cannot transition to a ferroelectric state due to topological protection of the domain wall network.

For valence change memory (VCM)‐type synapses, a large number of vacancies help to achieve very linearly changed dynamic range, and also, the low activation energy of vacancies enables low‐voltage operation. However, a large number of vacancies increases the current of artificial synapses by acting like dopants, which aggravates low‐energy operation and device scalability. Here, mixed‐dimensional formamidinium bismuth iodides featuring in‐situ formed type‐I band structure are reported for the VCM‐type synapse. As compared to the pure 2D and 0D phases, the mixed phase increases defect density, which induces a better dynamic range and higher linearity. In addition, the mixed phase decreases conductivity for non‐paths despite a large number of defects providing lots of conducting paths. Thus, the mixed phase‐based memristor devices exhibit excellent potentiation/depression characteristics with asymmetricity of 3.15, 500 conductance states, a dynamic range of 15, pico ampere‐scale current level, and energy consumption per spike of 61.08 aJ. A convolutional neural network (CNN) simulation with the Canadian Institute for Advanced Research‐10 (CIFAR‐10) dataset is also performed, confirming a maximum recognition rate of approximately 87%. This study is expected to lay the groundwork for future research on organic bismuth halide‐based memristor synapses usable for a neuromorphic computing system. Mixed‐dimensional formamidinium bismuth iodides exhibit highly linear potentiation/depression characteristics with energy consumption as low as 61.08 aJ for the convolutional neural network (CNN) due to a large number of defects and in‐situ formed type I band alignment.

Pelvic fixation is performed to obtain proper coronal and sagittal alignment when the pelvic obliquity is more than 15º in patients with neuromuscular scoliosis (NMS). Since many NMS patients were wheelchair or bed-ridden status, there has been controversy on the effect of pelvic fixation. Therefore, the purpose of this study is to analyze the effects of pelvic fixation on correction of spinal deformity and quality of life (QoL) in NMS patients. A total of 77 NMS patients who underwent deformity correction were divided into three groups and retrospectively analyzed preoperatively, postoperatively, and at 2-year follow-up: pelvic fixation group (Group A, n = 16), fixed to S1 (Group B, n = 33), and fixed to L5 (Group C, n = 28). The correction rate of scoliosis was 60.0%, 58.0%, and 56.7% in groups A, B, and C, respectively, with no statistical difference ( P  > 0.05). The correction rate of pelvic obliquity was 61.3%, 42.8%, and 57.5% in respective groups A, B, and C, with no significance ( P  > 0.05). The correction loss of scoliosis and pelvic obliquity showed no statistical significance between three groups for 2-year follow-up (all P s > 0.05). There were no significant differences regarding clinical outcomes and postoperative complications among the three groups (all P s > 0.05). Therefore, pelvic fixation using iliac screws is not substantially influencing radiological and clinical outcomes in the patients with NMS.

Perinatal outcomes of twin pregnancies are determined by several factors, such as gestational age (GA), chorionicity, and discordance at birth. This retrospective study aimed to investigate the association of chorionicity and discordance with neonatal and neurodevelopmental outcomes in preterm twin infants from uncomplicated pregnancy. Data of very preterm twin infants who were both live-born between 2014 and 2019 on the chorionicity of the twin, diagnosis of the twin-to-twin syndrome (TTTS), weight discordance at birth, and neonatal and neurodevelopmental outcomes at 24 months of corrected age (CA) were collected. Of the 204 twin infants analyzed, 136 were dichorionic (DC) and 68 were monochorionic (MC), including 15 pairs with TTTS. After adjusting for GA, brain injury, including severe intraventricular hemorrhage and periventricular leukomalacia, was mostly found in the MC with TTTS group, with a higher incidence of cerebral palsy and motor delay at CA 24 months. After excluding TTTS, multivariable analysis showed no association between chorionicity and neonatal and developmental outcomes, whereas small infants among co-twins (adjusted odds ratio (aOR) 3.33, 95% confidence interval 1.03–10.74) and greater discordance (%) of weight at birth (aOR 1.04, 1.00–1.07) were associated with neurodevelopmental impairment. Monochorionicity might not determine adverse outcomes among very preterm twins from uncomplicated pregnancy.