Search

Simple Summary: Isoorientin is an excellent antioxidant and regulator of lipid metabolism. We added this drug to the in vitro medium of porcine embryos and found a significant increase in the developmental rate and quality of the embryos. Isoorientin (ISO) is a natural lignan glycoside flavonoid found in various plants, including Charcot and Stonecrop. ISO exhibits diverse physiological and pharmacological effects, such as antioxidative, anti-inflammatory, hepatoprotective, antiviral, antianxiety, and anti-myocardial ischaemic properties, as well as lipid metabolism regulation. This study investigated the impact of ISO supplementation on oxidative stress and lipid accumulation in porcine early embryos, along with its underlying mechanisms. Porcine embryos were cultured in vitro under different concentrations of ISO (0, 1, 10, and 100 nM). The results revealed that 10 nM ISO significantly enhanced the blastocyst rate and total embryonic cell count in vitro. ISO-treated embryos exhibited reduced reactive oxygen species levels and elevated glutathione levels compared to the untreated group. In addition, ISO treatment significantly increased the expression of the key antioxidant regulator Nrf2, improved mitochondrial function, and reduced lipid droplet accumulation. Concurrently, early embryo autophagy and apoptosis levels decreased. Furthermore, ISO treatment upregulated antioxidant-related genes (SOD1, SOD2, and CAT) and mitochondrial biogenesis related genes (NRF1, NRF2, and SIRT1), while downregulating lipid synthesis-related genes (SREBP1 and FASN). Additionally, lipid hydrolysis-related genes (ACADS) were elevated. These findings collectively suggest that ISO may facilitate early embryonic development in pigs by ameliorating oxidative stress and lipid metabolism. [ABSTRACT FROM AUTHOR]

This study examines the impact of Notoginsenoside R1 (NGR1), a compound from Panax notoginseng, on the maturation of porcine oocytes and their embryonic development, focusing on its effects on antioxidant levels and mitochondrial function. This study demonstrates that supplementing in vitro maturation (IVM) medium with NGR1 significantly enhances several biochemical parameters. These include elevated levels of glutathione (GSH), nuclear factor erythrocyte 2‐related factor 2 (NRF2) and mRNA expression of catalase (CAT) and GPX. Concurrently, we observed a decrease in reactive oxygen species (ROS) levels and an increase in JC‐1 immunofluorescence, mitochondrial distribution, peroxisome proliferator‐activated receptor‐γ coactivator‐1α (PGC1α) and nuclear NRF2 mRNA levels. Additionally, there was an increase in ATP production and lipid droplets (LDs) immunofluorescence. These biochemical improvements correlate with enhanced embryonic outcomes, including a higher blastocyst rate, increased total cell count, enhanced proliferative capacity and elevated octamer‐binding transcription factor 4 (Oct4) and superoxide dismutase 2 (Sod2) gene expression. Furthermore, NGR1 supplementation resulted in decreased apoptosis, reduced caspase 3 (Cas3) and BCL2‐Associated X (Bax) mRNA levels and decreased glucose‐regulated protein 78 kD (GRP78) immunofluorescence in porcine oocytes undergoing in vitro maturation. These findings suggest that NGR1 plays a crucial role in promoting porcine oocyte maturation and subsequent embryonic development by providing antioxidant levels and mitochondrial protection. [ABSTRACT FROM AUTHOR]

Simple Summary: The in vitro maturation of oocytes is important for animal husbandry and assisted reproductive technology, but due to the higher oxidative stress in the in vitro environment compared with the in vivo environment, it is necessary to improve the efficiency of this process by alleviating in vitro oxidative stress. Eupatilin is a flavonoid compound with various activities, such as anti-inflammatory, anti-oxidant, and anti-tumoral activities. It is currently unclear whether it can reduce oxidative stress and improve the efficiency of the in vitro maturation of oocytes. The results of this study indicate that adding eupatilin to the in vitro maturation medium can alleviate oxidative stress, endoplasmic reticulum stress, and DNA damage in oocytes, further stimulating the maturation rate and embryo rate of porcine oocytes. It is expected to improve in vitro maturation efficiency and improve the composition of the medium. Eupatilin (5,7-dihydroxy-3′,4′,6-trimethoxyflavone) is a flavonoid derived from Artemisia plants that has beneficial biological activities, such as anti-apoptotic, anti-oxidant, and anti-inflammatory activities. However, the protective effects of eupatilin against oxidative stress and endoplasmic reticulum stress in porcine oocyte maturation are still unclear. To investigate the effect of eupatilin on the development of porcine oocytes after in vitro maturation and parthenogenetic activation, we added different concentrations of eupatilin in the process of porcine oocyte maturation in vitro, and finally selected the optimal concentration following multiple comparisons and analysis of test results using SPSS (version 17.0; IBM, Chicago, IL, USA) software. The results showed that 0.1 μM eupatilin supplementation did not affect the expansion of porcine cumulus cells, but significantly increased the extrusion rate of porcine oocyte polar bodies, the subsequent blastocyst formation rate, and the quality of parthenogenetically activated porcine embryos. Additionally, it reduced the level of reactive oxygen species in cells and increased glutathione production. Further analysis revealed that eupatilin supplementation could reduce apoptosis, DNA double-strand breaks, and endoplasmic reticulum stress. In conclusion, supplementation with 0.1 μM eupatilin during in vitro maturation improved oocyte maturation and subsequent embryo development by reducing oxidative stress and endoplasmic reticulum stress. [ABSTRACT FROM AUTHOR]

Our previous study established that chrysoeriol (CHE) can reduce reactive oxygen species (ROS) accumulation, apoptosis, and autophagy in vitro culture (IVC) of porcine embryos. However, the role of CHE in oocyte maturation and lipid homeostasis is unclear. Herein, we aimed to elucidate the effect of CHE on porcine oocyte competence in vitro maturation (IVM) and subsequent embryo development. The study chooses parthenogenetic activated porcine oocytes as the research model. The study revealed that the cumulus expansion index and related gene expressions are significantly elevated after supplementing 1 μM CHE. Although there were no significant differences in nuclear maturation and cleavage rates, the blastocyst formation rate and total cell numbers were significantly increased in the 1 μM CHE group. In addition, CHE improved the expression of genes related to oocyte and embryo development. ROS was significantly downregulated in all CHE treatment groups, and intracellular GSH (glutathione) was significantly upregulated in 0.01, 0.1, and 1 μM CHE groups. The immunofluorescence results indicated that mitochondrial membrane potential (MMP) and lipid droplet (LD), fatty acid (FA), ATP, and functional mitochondria contents significantly increased with 1 μM CHE compared to the control. Furthermore, CHE increased the expression of genes related to lipid metabolism, mitochondrial biogenesis, and β-oxidation. [ABSTRACT FROM AUTHOR]

Notoginsenoside R1 (NGR1), derived from the Panax notoginseng root and rhizome, exhibits diverse pharmacological influences on the brain, neurons, and osteoblasts, such as antioxidant effects, mitochondrial function protection, energy metabolism regulation, and inhibition of oxygen radicals, apoptosis, and cellular autophagy. However, its effect on early porcine embryonic development remains unclear. Therefore, we investigated NGR1's effects on blastocyst quality, reactive oxygen species (ROS) levels, glutathione (GSH) levels, mitochondrial function, and embryonic development‐related gene expression in porcine embryos by introducing NGR1 during the in vitro culture (IVC) of early porcine embryos. Our results indicate that an addition of 1 μM NGR1 significantly increased glutathione (GSH) levels, blastocyst formation rate, and total cell number and proliferation capacity; decreased ROS levels and apoptosis rates in orphan‐activated porcine embryos; and improved intracellular mitochondrial distribution, enhanced membrane potential, and reduced autophagy. In addition, pluripotency‐related factor levels were elevated (NANOG and octamer‐binding transcription factor 4 [OCT4]), antioxidant‐related genes were upregulated (nuclear factor‐erythroid 2‐related factor 2 [NRF2]), and apoptosis‐ (caspase 3 [CAS3]) and autophagy‐related genes (light chain 3 [LC3B]) were downregulated. These results indicate that NGR1 can enhance early porcine embryonic development by protecting mitochondrial function. [ABSTRACT FROM AUTHOR]

Summary: Mammalian oocytes not fertilized immediately after ovulation can undergo ageing and a rapid decline in quality. The addition of antioxidants can be an efficient approach to delaying the oocyte ageing process. Onion peel extract (OPE) contains quercetin and other flavonoids with natural antioxidant activities. In this study, we investigated the effect of OPE on mouse oocyte ageing and its mechanism of action. The oocytes were aged in vitro in M16 medium for 16 h after adding OPE at different concentrations (0, 50, 100, 200, and 500 μg/ml). The addition of 100 μg/ml OPE reduced the oocyte fragmentation rate, decreased the reactive oxygen species (ROS) level, increased the glutathione (GSH) level, and improved the mitochondrial membrane potential compared with the control group. The addition of OPE also increased the expression of SOD1 , CAT , and GPX3 genes, and the caspase-3 activity in OPE-treated aged oocytes was significantly lower than that in untreated aged oocytes and similar to that in fresh oocytes. These results indicated that OPE delayed mouse oocyte ageing by reducing oxidative stress and apoptosis and enhancing mitochondrial function. [ABSTRACT FROM AUTHOR]

Brain organoids have been considered as an advanced platform for in vitro disease modeling and drug screening, but numerous roadblocks exist, such as lack of large-scale production technology and lengthy protocols with multiple manipulation steps, impeding the industrial translation of brain organoid technology. Here, we describe the high-speed and large-scale production of midbrain organoids using a high-throughput screening-compatible platform within 30 days. Micro midbrain organoids (µMOs) exhibit a highly uniform morphology and gene expression pattern with minimal variability. Notably, µMOs show dramatically accelerated maturation, resulting in the generation of functional µMOs within only 30 days of differentiation. Furthermore, individual µMOs display highly consistent responsiveness to neurotoxin, suggesting their usefulness as an in vitro high-throughput drug toxicity screening platform. Collectively, our data indicate that µMO technology could represent an advanced and robust platform for in vitro disease modeling and drug screening for human neuronal diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: The residue of chemicals can affect the maturation of livestock oocytes, thereby affecting the quality of embryos. Research has shown that imidacloprid, a systemic neonicotinoid insecticide widely used in agriculture, can damage the reproductive system of mammals, and it is unclear whether imidacloprid will affect oocyte maturation and how to reduce its toxicity. The results of this study show that imidacloprid can affect oocyte maturation, and melatonin supplementation can restore oocyte maturation by alleviating oxidative stress and endoplasmic reticulum stress, and offset the toxic effect of imidacloprid on pig oocytes, which indicates that Melatonin may be a promising drug to improve the quality of oocytes exposed to imidacloprid in animals. Imidacloprid (IMI) is an endogenous neonicotinoid insecticide widely used in agriculture and has attracted researchers' attention because of its risks to the environment and human health. Melatonin (MT) is an antioxidant hormone produced by the pineal gland of the brain. Studies have shown that it has a variety of physiological functions and plays a crucial role in the development of animal germ cells and embryos. The potential protective effects of MT against oocyte damage caused by neonicotinoid pesticide toxicity remain unclear. In this study, we report the toxicity of IMI against, and its effects on the quality of, porcine oocytes and the protective effect of MT on IMI-exposed oocytes. The results show that IMI exposure adversely affected oocyte maturation, while MT supplementation ameliorated its toxic effects. Specifically, IMI exposure increased oxidative stress (OS), endoplasmic reticulum stress (ERS), and apoptosis, which may affect polar body expulsion rates and blastocyst formation. Also, IMI exposure reduced oocyte cleavage rates and the number of cells in blastocysts. However, all of these toxic effects can be restored after a melatonin supplementation treatment. In conclusion, these results suggest that melatonin has a protective effect on IMI-induced defects during porcine oocyte maturation. [ABSTRACT FROM AUTHOR]

Simple Summary: The 2,2′4,4′-tetrabromodiphenyl ether (BDE-47) is a common flame retardant that can be widely distributed in the environment and organisms but has been shown to induce toxicity to various organisms and organ systems. We found that exposure to BDE-47 induced the early embryonic development of in vitro porcine culture through oxidative stress and autophagy induced by mitochondrial dysfunction and endoplasmic reticulum stress. Widely used as a flame retardant, 2,2′4,4′-tetrabromodiphenyl ether (BDE-47) is a persistent environmental pollutant with toxicological effects, including hepatotoxicity, neurotoxicity, reproductive toxicity, and endocrine disruption. To investigate the toxicological effects of BDE-47 on early porcine embryogenesis in vitro, cultured porcine embryos were exposed to BDE-47 during early development. Exposure to 100 μM BDE-47 decreased the blastocyst rate and mRNA level of pluripotency genes but increased the level of LC3 and the expression of autophagy-related genes. After BDE-47 exposure, porcine embryos' antioxidant capability decreased; ROS levels increased, while glutathione (GSH) levels and the expression of antioxidant-related genes decreased. In addition, BDE-47 exposure reduced mitochondrial abundance and mitochondrial membrane potential levels, downregulated mitochondrial biogenesis-associated genes, decreased endoplasmic reticulum (ER) abundance, increased the levels of GRP78, a marker of ER stress (ERS), and upregulated the expression of ERS-related genes. However, ER damage and low embryo quality induced by BDE-47 exposure were reversed with the ERS inhibitor, the 4-phenylbutyric acid. In conclusion, BDE-47 inhibits the development of early porcine embryos in vitro by inducing mitochondrial dysfunction and ERS. This study sheds light on the mechanisms of BDE-47-induced embryonic toxicity. [ABSTRACT FROM AUTHOR]

Modelings of the interface distribution and flow-induced residual stresses and birefringence in the sequential co-injection molding (CIM) of a center-gated disk were carried out using a numerical scheme based on a hybrid finite element/finite difference/control volume method. A nonlinear viscoelastic constitutive equation and stress-optical rule were used to model the frozen-in flow stresses in disks. The compressibility of melts is included in modeling of the packing and cooling stages and not in the filling stage. The thermally induced residual birefringence was calculated using the linear viscoelastic and photoviscoelastic constitutive equations combined with the first-order rate equation for volume relaxation and the master curves for the relaxation modulus and strain-optical coefficient functions of each polymer. The influence of the processing variables including melt and mold temperatures and volume of skin melt on the birefringence and interface distribution was analyzed for multi-layered PS-PC-PS, PS-PMMA-PS, and PMMA-PC-PMMA molded disks obtained by CIM. The interface distribution and residual birefringence in the molded disks were measured. The measured interface distributions and the gapwise birefringence distributions in CIM disks were found to be in a fair agreement with the predicted interface distributions and the total residual birefringence obtained by the summation of the predicted frozen-in flow and thermal birefringence., INTRODUCTION The co-injection process is known since its first introduction in the early 1970s. In co-injection molding (CIM), two different (dissimilar but chemically and Theologically compatible) polymers are injected into [...]

Simple Summary: Excellent quality of early embryonic development contributes to a successful pregnancy. At present, most in vitro cultured embryos can only develop to the blastocyst stage at most, because an in vitro culture (IVC) system cannot replace the physiological environment in vivo. During IVC, excessive accumulated reactive oxygen species in embryos cannot be easily metabolized, which will cause oxidative stress and suppress embryo development. In this study, we found that anti-oxidation capacity of early embryo was improved by adding 6-gingerol to IVC. Moreover, 6-gingerol can also improve blastocyst rate, cell proliferation, mitochondrial function, inhibit cell apoptosis, autophagy, and regulate functional genes expression in blastocyst. These results are helpful to optimize the early embryo culture system, and thus provide a theoretical basis for improving the early embryo quality and the efficiency of subsequent pregnancy. 6-Gingerol, the main active ingredient in ginger, exhibits a variety of biological activities, such as antioxidant, anti-inflammatory, and anticancer activities, and can affect cell development. However, the effects of 6-gingerol on mammalian reproductive processes, especially early embryonic development, are unclear. This study explored whether 6-gingerol can be used to improve the quality of in vitro-cultured porcine embryos. The results showed that 5 μM 6-gingerol significantly increased the blastocyst formation rates of porcine early embryos. 6-Gingerol attenuated intracellular reactive oxygen species accumulation and autophagy, increased intracellular glutathione levels, and increased mitochondrial activity. In addition, 6-gingerol upregulated NANOG, SRY-box transcription factor 2, cytochrome c oxidase subunit II, mechanistic target of rapamycin kinase, and RPTOR independent companion of MTOR complex 2 while downregulating Caspase 3, baculoviral IAP repeat containing 5, autophagy related 12, and Beclin 1. Most importantly, 6-gingerol significantly increased the levels of p-extracellular regulated protein kinase 1/2 while reducing the levels of p-c-Jun N-terminal kinase 1/2/3 and p-p38. These results indicate that 6-gingerol can promote the development of porcine early embryos in vitro. [ABSTRACT FROM AUTHOR]

Chimeric lysins composed of various combinations of cell wall-lysing (enzymatic) and cell-wall-binding (CWB) domains of endolysins, autolysins, and bacteriocins have been developed as alternatives to or adjuvants of conventional antibiotics. The screening of multiple chimeric lysin candidates for activity via E. coli expression is not cost effective, and we previously reported on a simple cell-free expression system as an alternative. In this study, we sufficiently improved upon this cell-free expression system for use in screening activity via a turbidity reduction test, which is more appropriate than a colony reduction test when applied in multiple screening. Using the improved protocol, we screened and compared the antibacterial activity of chimeric lysin candidates and verified the relatively strong activity associated with the CHAP (cysteine, histidine-dependent amidohydrolase/peptidase) domain of secretory antigen SsaA-like protein (ALS2). ALS2 expressed in E. coli showed two major bands, and the smaller one (subprotein) was shown to be expressed by an innate downstream promoter and start codon (ATG). The introduction of synonymous mutations in the promoter resulted in clearly reduced expression of the subprotein, whereas missense mutations in the start codon abolished antibacterial activity as well as subprotein production. Interestingly, most of the S. aureus strains responsible for bovine mastitis were susceptible to ALS2, but those from human and chicken were less susceptible. Thus, the simple and rapid screening method can be applied to select functional chimeric lysins and define mutations affecting antibacterial activity, and ALS2 may be useful in itself and as a lead molecule to control bovine mastitis. [ABSTRACT FROM AUTHOR]

Simple Summary: In this experiment, porcine embryos were used as a model, and the flavonoid chrysoeriol (CHE) was added to the in vitro culture of the porcine embryo. Then, the effect of CHE on embryo development was studied by microscopic observation, immunofluorescence staining, and quantitative Real-time PCR (qRT-PCR). The results showed that CHE could reduce the accumulation of reactive oxygen species (ROS) and the level of oxidative stress during the development of porcine embryos, thus improving the antioxidant activity of the embryos. We also found that CHE can increase the mitochondrial membrane potential in the embryos and protect mitochondrial function. Further experiments preliminarily showed that CHE can reduce apoptosis and autophagy during embryonic development. We have proved that CHE has good antioxidant activity, and this is the first time that CHE has been used in mammalian embryo culture in vitro. The results show that CHE can improve the development of pig embryos in vitro by reducing oxidative stress and autophagy. Chrysoeriol (CHE) is a flavonoid substance that exists in many plants. It has various physiological and pharmacological effects, including anti-inflammatory, antioxidant, anti-tumor, and protective activity, especially for the cardiovascular system and liver. Among common livestock embryos, porcine embryos are often considered high-quality objects for studying the antioxidant mechanisms of oocytes. Because porcine embryos contain high levels of lipids, they are more vulnerable to external stimuli, which affect development. Our study explored the influence of CHE supplementation on oxidative stress in porcine oocytes and its possible mechanisms. Different concentrations of CHE (0, 0.1, 1, and 3 µM) were supplemented in the in vitro culture medium of the porcine oocytes. The results showed that supplementation with 1 µM CHE significantly increased the blastocyst rate and total cell number of embryos in vitro. After finding the beneficial effects of CHE, we measured reactive oxygen species (ROS), glutathione (GSH), and mitochondrial membrane potential (MMP) when the oocytes reached the 4-cell stage of development and determined the levels of apoptosis, cell proliferation, and autophagy at the blastocyst stage of development. The expression levels of some related genes were preliminarily detected by qRT-PCR. The results showed that the apoptosis of blastocysts in the CHE-treated culture also decreased compared with the untreated culture. Furthermore, CHE downregulated intracellular ROS and increased GSH in the embryos. CHE was also shown to improve the activity of mitochondria and inhibit the occurrence of autophagy. In addition, antioxidant-related genes (SOD1, SOD2, and CAT) and cell pluripotency-related genes (SOX2, OCT4, and NANOG) were upregulated. At the same time, apoptosis-related (Caspase 3) and autophagy-related (LC3B) genes showed a downward trend after supplementation with CHE. These results indicate that CHE improved the development of porcine embryos in vitro by reducing oxidative stress and autophagy levels. [ABSTRACT FROM AUTHOR]

(1) Background: DNA double strand breaks (DSBs) are the most serious form of DNA damage that affects oocyte maturation and the physiological state of follicles and ovaries. Non-coding RNAs (ncRNAs) play a crucial role in DNA damage and repair. This study aims to analyze and establish the network of ncRNAs when DSB occurs and provide new ideas for next research on the mechanism of cumulus DSB. (2) Methods: Bovine cumulus cells (CCs) were treated with bleomycin (BLM) to construct a DSB model. We detected the changes of the cell cycle, cell viability, and apoptosis to determine the effect of DSBs on cell biology, and further evaluated the relationship between the transcriptome and competitive endogenous RNA (ceRNA) network and DSBs. (3) Results: BLM increased γH2AX positivity in CCs, disrupted the G1/S phase, and decreased cell viability. Totals of 848 mRNAs, 75 long noncoding RNAs (lncRNAs), 68 circular RNAs (circRNAs), and 71 microRNAs (miRNAs) in 78 groups of lncRNA–miRNA–mRNA regulatory networks, 275 groups of circRNA–miRNA–mRNA regulatory networks, and five groups of lncRNA/circRNA–miRNA–mRNA co-expression regulatory networks were related to DSBs. Most differentially expressed ncRNAs were annotated to cell cycle, p53, PI3K-AKT, and WNT signaling pathways. (4) Conclusions: The ceRNA network helps to understand the effects of DNA DSBs activation and remission on the biological function of CCs. [ABSTRACT FROM AUTHOR]

The simulation of the gapwise distribution of the thermally-induced residual birefringence and stresses in freely-quenched PS-PC-PS and PC-PS-PC multi-layered slabs in water was carried out to calculate the gapwise distribution of the transient and residual birefringence. The modeling was based on the linear viscoelastic and photoviscoelastic constitutive equations combined with the first-order rate equation for volume relaxation. The master curves for the Young's relaxation modulus and strain-optical coefficient functions obtained earlier for PS and PC were used in the simulations. The obtained numerical results provided the evolution of the thermally-induced stress and birefringence with time during and after quenching. The predicted gapwise residual birefringence distribution in these slabs was found to be in a fair agreement with the measured results. In addition, the gapwise distribution of the thermally-induced residual birefringence in the multi-layered PS-PMMA-PS, PMMA-PS-PMMA, PMMA-PC-PMMA, and PC-PMMA-PC slabs quenched from different initial temperatures was measured. Explanations were provided for the observed gapwise distribution of the thermal residual birefringence in each layer of these slabs including the effect of the initial temperature., INTRODUCTION During polymer processing, the polymer undergoes simultaneous mechanical and thermal influences in the fluid, rubbery, and glassy states. These effects introduce orientation, residual stresses, and shrinkage in the products [...]