RESUMO
The quality of pre-implantation embryos could affect developmental efficiency after embryo transfer. However, the assessment of pre-implantation embryos was unsatisfactory, especially in pig embryos to date. Therefore, this study was designed to investigate available and applicable parameters that indicate developmental potential and quality of porcine pre-implantation embryos produced by handmade cloning (HMC), and parthenogenetic activation without zona pellucida (PAZF) and with zona pellucida (PAZI). Firstly, a common division behaviour was detected, that is the formation of uneven division with two unequal size blastomeres (UD 2-cell), especially in HMC embryos; then, the proportion of UD 2-cell was found to be significantly higher than that of even division with equal size blastomeres (ED 2-cell) (72.56 ± 4.56 vs. 24.57 ± 1.92). The formation of UD 2-cell might be due to the spindle migration along the long axis in 1-cell stage, and the cleavage furrow was not formed in the centre of cytoplasm. In the two sister blastomeres of UD 2-cell, uneven distribution of organelles (mitochondria and lipid droplet) was observed with lower proportion in the smaller one (p < .05). Although no difference in blastocyst rate was observed between UD and ED 2-cell embryos, the cell number per blastocyst from UD 2-cell embryos was lower than that from ED 2-cell embryos (44.15 ± 2.05 vs. 51.55 ± 1.83). Besides, because of non-synchronized division of each blastomere, the following three cleavage routes were observed in all HMC/PAZF/PAZI embryos: T1 (2-cell â 3-cell â 4-cell â ≥5-cell â morula â blastocyst), T2 (2-cell â 3-cell â 4-cell â morula â blastocyst) and T3 (2-cell â 3-cell/4-cell â morula â blastocyst). Therefore, in pig in vitro-produced embryos, division behaviours of uneven volume of cytoplasm and non-synchronized cell cycles were observed at the early embryonic developmental stage, which might be another potential factor to evaluate embryonic development.
Assuntos
Blastômeros , Desenvolvimento Embrionário , Animais , Blastocisto , Feminino , Mórula , Partenogênese , Gravidez , SuínosRESUMO
Increased palmitic acid (PA) levels have been found in females with reduced fertility due to metabolic disorders. However, effective antioxidant astaxanthin (AXE) might positively affect animal reproduction. Therefore, the present study was designed to evaluate the impact of a high concentration of PA on oocyte maturation and the possible protective effect of AXE against high PA concentration in pigs. Firstly, different concentrations (0.2, 0.5, 0.8 mM) of PA were conducted on in vitro maturation (IVM) of pig oocytes (PA0.2, PA0.5, and PA0.8), while no addition of PA was performed as the control group (Ctrl). Results showed that the cumulus cell expansion index (CCEI) was lower in PA0.5 and PA0.8 groups compared to Ctrl group (p < .05). In PA0.5 group, not only did the percentage of matured oocytes with the first polar body (PB1) reduced, that with more oocytes arrested at germinal vesicle (GV) stage (53.44% ± 7.16% vs. 20.93% ± 5.16%, p < .05), but also a higher number of transzonal projections (TZPs) was observed in PA0.5 than Ctrl group. Besides, supplement of PA resulted in a dose-dependent decrease in mitochondrial activity. Although no difference of lipid content was observed between PA0.5 and Ctrl groups, the lipid content was at a higher level in PA0.2 group than in the other three groups. Hence, concentration of 0.5 mM of PA was performed in the following experiments, and 2.5 µM AXE carried out to investigate the possible relief effects of PA (PA0.5 + AXE). Results showed that the percentage of matured oocytes with PB1 was higher in PA0.5 + AXE than in PA0.5 group (63.43% ± 1.50% vs. 55.33% ± 0.81%, p < .01), and ROS levels both in oocytes and their cumulus cells (CCs) reduced in PA0.5 + AXE when compared to PA0.5 group. In addition, the rate of CCs with apoptosis decreased in PA0.5 + AXE, and the expression level of caspase 3 and BAX was lower than PA0.5 group. In conclusion, the maturation of pig oocytes was inhibited by the high concentration of PA; however, this negative effect of PA-induced might be relieved by the supplement of AXE.
Assuntos
Técnicas de Maturação in Vitro de Oócitos , Ácido Palmítico , Feminino , Animais , Suínos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Ácido Palmítico/farmacologia , Ácido Palmítico/metabolismo , Células do Cúmulo , OócitosRESUMO
To address the non-negligible volume expansion and the inherent poor electronic conductivity of silica (SiO2 ) material, microsphere-like SiO2 /MXene hybrid material is designed and successfully synthesized through the combination of the Stöber method and spray drying. The SiO2 nanoparticles are firmly anchored on the laminated MXene by the bonding effect, which boosts the structural stability during the long-term cycling process. The MXene matrix not only possesses high elasticity to buffer the volume variation of SiO2 nanoparticles, but also promotes the transfer of electrons and lithium ions. Moreover, the microsphere wrapped with ductile MXene film reduces the specific surface area, relieves the side reactions, and enhances the coulombic efficiency. Therefore, superior electrochemical performance including high reversible capacity, outstanding cycle stability, high coulombic efficiency, especially in the first cycle, excellent rate capability as well as high areal capacity are acquired for SiO2 /MXene microspheres anode.
RESUMO
The interaction between Li0.33La0.56TiO3 (LLTO) and metallic lithium leads to severe interfacial instability of LLTO-containing solid-state electrolytes with a lithium metal anode. To improve the interfacial stability, a heterogeneous composite electrolyte PVDF-HFP@LLTO/PEO (PLTP) is designed and fabricated with a PEO electrolyte layer adhered to the PVDF-HFP@LLTO (PLT) electrolyte membrane. The PLTP heterogeneous composite electrolyte exhibits a superior ionic conductivity of 3.23 × 10-4 S cm-1 at 60 °C and a highly stable electrochemical window of up to 4.7 V (vs. Li/Li+). Remarkably, taking advantage of the effective protection of the PEO electrolyte layer, the chemical stability at the electrolyte/lithium metal anode interface is significantly enhanced. As a result, solid-state Li||LiFePO4 and Li||LiNi0.6Co0.2Mn0.2O2 batteries with the heterogeneous electrolyte exhibit an impressive electrochemical performance with high Coulombic efficiency and stable cycling capability. The strengthened interfacial stability enables the heterogeneous electrolyte to be a promising alternative for the further development of solid-state lithium metal batteries.
RESUMO
Devising synthetic strategies to control crystal structure is of great importance as materials properties are governed by structure. MnS is a great model system as it has three known stable polymorphs. Herein, we show the selective synthesis of colloidal wurtzite- and rock-salt-type MnS under identical reactions conditions changing only the manganese halide precursor. Mixtures of Mn halides or halide surrogate (e.g., NH4Cl) also enabled polymorph control. Powder X-ray diffraction aliquot studies of the reactions revealed the crystal structure at the onset of nucleation and that of the final product is the same, unlike the Ostwald ripening transformation observed in other systems. The halide-driven selectivity was also observed in the synthesis of manganese selenide nanoparticles. In this system, variation of the Mn halide precursor allowed access to the wurtzite- and rock salt-type polymorphs of MnSe, as well as the pyrite-MnSe2 phase. Based on this work, the mixing of metal salts might be a simple and effective strategy towards polymorph control and access materials with new crystal structures.
RESUMO
Bromodomain PHD finger transcription factor (BPTF), a core subunit of nucleosome-remodeling factor (NURF) complex, plays an important role in chromatin remodeling. However, few information of BPTF is available in pig, especially in mammalian follicular granulosa cells (GCs). The present study firstly confirmed that BPTF in porcine was relative close to human and mouse. The expression of BPTF could be detected in ovary, testes, lung, kidney, large intestine, and small intestine. And a relative high expression of BPTF was observed in ovarian follicles and GCs. When BPTF was knocked down (BPTF-siRNA), the viability of GCs was affected. And the expression level of CDK1, cyclin B1, CDK4 and CDK2 was higher than the control, which might indicate that the cell cycle of GCs was inhibited from S to G2/M phase. Although the apoptosis level was induced in the BPTF-siRNA GCs, the reduced level of H3K4 methylation was detected with the down regulation of SMYD3, EHMT2 and DPY30. Thereby, results in the present might provide the primary knowledge of BPTF in GCs and the follicular development in pig.
Assuntos
Montagem e Desmontagem da Cromatina , Células da Granulosa , Feminino , Humanos , Suínos , Animais , Camundongos , Ciclo Celular , Divisão Celular , Apoptose , Mamíferos , Antígenos de Histocompatibilidade , Histona-Lisina N-MetiltransferaseRESUMO
Epigenetic status of fetal fibroblasts (FFs) is one of the crucial factors accounted for the success of somatic cell nuclear transfer and gene editing, which might inevitably be affected by passaging. But few systematic studies have been performed on the epigenetic status of passaged aging cells. Therefore, FFs from large white pig were in vitro passaged to the 5, 10, and 15 (F5, F10, and F15) passages in the present study to investigate the potential alteration of epigenetic status. Results indicated the senescence of FFs occurs with the passaging, as assessed by the weakened growth rate, increased ß-gal expression, and so on. For the epigenetic status of FFs, the higher level both of DNA methylation and H3K4me1, H3K4me2, H3K4me3 was observed at F10, but the lowest level was observed at F15. However, the fluorescence intensity of m6A was significantly higher in F15, but lower (p < 0.05) in F10, and the related mRNA expression in F15 was significantly higher than F5. Further, RNA-Seq indicated a considerable difference in the expression pattern of F5, F10, and F15 FFs. Among differentially expressed genes, not only the genes involved in cell senescence were changed, but also the upregulated expression of Dnmt1, Dnmt3b, Tet1 and dysregulated expression of histone methyltransferases-related genes were detected in F10 FFs. In addition, most genes related to m6A such as METTL3, YTHDF2, and YTHDC1 were significantly different in F5, F10, and F15 FFs. In conclusion, the epigenetic status of FFs was affected by being passaged from F5 to F15.
Assuntos
Metilação de DNA , Epigênese Genética , Suínos , Animais , Metilação de DNA/genética , Técnicas de Transferência Nuclear , Fibroblastos/metabolismoRESUMO
Different development stages of porcine embryos have different tolerance to low temperature. Therefore, we took the porcine embryos after parthenogenetic activation (PA) as the model, to explore the optimal development stage for vitrification during morula (D4), early blastocyst (D5), and expanded blastocyst (D6) after PA (D0). Embryos were observed with microscope and analyzed by different staining after cryo-recovery for 24 hours. The quality of embryos was damaged after vitrification, including embryonic nuclei, DNA, cytoskeleton, and organelles. The re-expansion rate at 24 hours of D5 embryos was significantly higher than those of D4 and D6 embryos (D5 vs. D4 vs. D6, 27.620 ± 0.041 vs. 7.809 ± 0.027 vs. 13.970 ± 0.032, p < 0.05). Therefore, D5 embryos were selected as research objects to explore the effect of vitrification on lipid in vitrified embryos. The results showed that the expression levels of perilipin PLIN3 messenger RNA (mRNA) and triacylglycerol synthesis-related genes AGPAT1 and DGAT mRNA are significantly reduced (p < 0.05). Vitrification affected lipid synthesis, which might have an irreversible impact on embryonic development. In conclusion, our data demonstrated that the optimal stage of vitrification was D5 for early blastocysts.
Assuntos
Criopreservação , Vitrificação , Animais , Blastocisto/fisiologia , Criopreservação/métodos , Criopreservação/veterinária , Desenvolvimento Embrionário/fisiologia , Feminino , Lipídeos , Gravidez , RNA Mensageiro , SuínosRESUMO
An amount (5 wt %) of lithium tungstate (Li2WO4) as an additive significantly improves the cycle and rate performances of the LiNi0.6Co0.2Mn0.2O2 electrode at the cutoff voltage of 4.6 V. The 5 wt % Li2WO4-mixed LiNi0.6Co0.2Mn0.2O2 electrode delivers a reversible capacity of 199.2 mA h g-1 and keeps 73.1% capacity for 200 cycles at 1 C. It retains 67.4% capacity after 200 cycles at 2 C and delivers a discharge capacity of 167.3 mA h g-1 at 10 C, while those of the pristine electrode are only 44.7% and 87.5 mA h g-1, respectively. It is shown that the structure of the LiNi0.6Co0.2Mn0.2O2 cathode material is not affected by mixing Li2WO4. The introduced Li2WO4 effectively restrains the LiPF6 and carbonate solvent decomposition by consuming PF5 at high cutoff voltage, forming a stable cathode/electrolyte interface film with low resistance.