Effect of vitrification on protein O-GlcNAcylation in mouse metaphase II oocytes.

In brief: Oocyte vitrification leads to DNA hypomethylation, which results in defect in early embryo development. This study reveals that oocyte vitrification impairs the DNA methylation pattern by influencing protein O-GlcNAcylation. Abstract: Oocyte vitrification leads to decreased DNA methylation levels, which impairs the quality and the developmental potential of oocytes. However, the underlying molecular mechanism still need to be further revealed. In this study, mouse metaphase II (M II) oocytes were frozen by vitrification technology, while fresh oocytes were used as the control group. The effect of oocyte vitrification on protein O-GlcNAcylation and its impact on the developmental potential of oocytes were elucidated. We found that the protein O-GlcNAcylation levels were significantly increased in vitrified oocytes. Increase of protein O-GlcNAcylation levels in control oocytes by PUGNAc (an O-GlcNAcase inhibitor) decreases blastocyst rate after parthenogenetic activation (20.82% in PUGNAc-treated group; 53.82% in control group, P < 0.05). We also discovered that DNA methylation was disrupted in two-cell embryos derived from vitrified oocytes, resulting in decreased 5mC and increased 5hmC, showing similar phenotypes to that derived from PUGNAc-treated oocytes. In vitrified and PUGNAc-treated oocytes, O-GlcNAcylated TET3 was significantly increased. Notably, by inhibiting protein O-GlcNAcylation in vitrified oocytes using OSMI1 (an O-GlcNAc transferase inhibitor) we restored the DNA methylation in two-cell embryos and ameliorated the developmental defects in early embryo. Thus, elevated protein O-GlcNAcylation in vitrified oocytes is an essential contributor to their declining embryonic developmental potential. Modulation of protein O-GlcNAcylation improves the developmental potential of vitrified oocytes.

Mechanisms and effects of novel ammonifying microorganisms on nitrogen ammonification in cow manure waste composting.

It is essential to reduce nitrogen losses and to improve nitrogen conversion during organic waste composting because of environmental protection and sustainable development. To reveal newly domesticated ammonifying microorganisms (AM) cultures on the ammonification and nitrogen conversion during the composting, the screened microbial agents were inoculated at 5 % concentration (in weight basis) into cow manure compost under five different treatments: sterilized distilled water (Control), Amm-1 (mesophilic fungus-F1), Amm-2 (mesophilic bacterium-Z1), Amm-3 (thermotolerant bacterium-Z2), and Amm-4 (consortium: F1, Z1, and Z2), and composted for 42 days. Compared to control, AM inoculation prolonged the thermophilic phases to 9-19 days, increased the content of NH4+-N to 1.60-1.96 g/kg in the thermophilic phase, reduced N2O and NH3 emissions by 22.85-61.13 % and 8.45-23.29 %, increased total Kjeldahl nitrogen, and improved cell count and viability by 12.09-71.33 % and 66.71-72.91 %. AM was significantly associated with different nitrogen and microbial compositions. The structural equation model (SEM) reveals NH4+-N is the preferable nitrogen for the majority of bacterial and fungal growth and that AM is closely associated with the conversion between NH3 and NH4+-N. Among the treatments, inoculation with Amm-4 was more effective, as it significantly enhanced the driving effect of the critical microbial composition on nitrogen conversion and accelerated nitrogen ammonification and sequestration. This study provided new concepts for the dynamics of microbial in the ammonification process of new AM bacterial agents in cow manure compost, and an understanding of the ecological mechanism underlying the ammonification process and its contribution to nitrogen (N) cycling from the perspective of microbial communities.

Occurrence, ecology risk assessment and exposure evaluation of 19 anthelmintics in dust and soil from China.

In order to fill the blank of domestic research on anthelmintics in dust and soil, 159 paired dust (including indoor and outdoor dust) and soil samples were collected nationwide. All 19 kinds of the anthelmintics were detected in the samples. The total concentration of the target substances in the outdoor dust, indoor dust and soil samples ranged from 1.83 to 1.30 × 103 ng/g, from 2.99 to 6.00 × 103 ng/g and from 0.23 to 8.03 × 102 ng/g, respectively. The total concentration of the 19 anthelmintics in northern China were significantly higher than those in southern China in the outdoor dust and soil samples. No significant correlation was found in the total concentration of anthelmintics between the indoor and outdoor dust because of strong human activities interference, however, a significant correlation existed between the outdoor dust and soil samples and between the indoor dust and soil samples. High ecological risk was found at 35% and 28% of all the sampling sites to non-target organisms in the soil respectively for IVE and ABA, and merits further study. The daily anthelmintics intakes were evaluated via ingestion and dermal contact of soil and dust samples for both children and adults. Ingestion was the predominant way for anthelmintics exposure, and the anthelmintics in soil and dust did not pose a health threat to human health at present.

Alkaline Dilution Alters Sperm Motility in Dairy Goat by Affecting sAC/cAMP/PKA Pathway Activity.

In dairy goat farming, increasing the female kid rate is beneficial to milk production and is, therefore, economically beneficial to farms. Our previous study demonstrated that alkaline incubation enriched the concentration of X-chromosome-bearing sperm; however, the mechanism by which pH affects the motility of X-chromosome-bearing sperm remains unclear. In this study, we explored this mechanism by incubating dairy goat sperm in alkaline dilutions, examining the pattern of changes in sperm internal pH and Ca2+ concentrations and investigating the role of the sAC/cAMP/PKA pathway in influencing sperm motility. The results showed that adding a calcium channel inhibitor during incubation resulted in a concentration-dependent decrease in the proportion of spermatozoa with forward motility, and the sperm sAC protein activity was positively correlated with the calcium ion concentration (r = 0.9972). The total motility activity, proportion of forward motility, and proportion of X-chromosome-bearing sperm decreased (p < 0.05) when cAMP/PKA protease activity was inhibited. Meanwhile, the enrichment of X-chromosome-bearing sperm by pH did not affect the sperm capacitation state. These results indicate that alkaline dilution incubation reduces Ca2+ entry into X-sperm and the motility was slowed down through the sAC/cAMP/PKA signaling pathway, providing a theoretical foundation for further optimization of the sex control method.

Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes.

BACKGROUND: Oocyte vitrification has been widely used in the treatment of infertility and fertility preservation. However, vitrification-induced mitochondrial damage adversely affects oocyte development. Several studies have reported that mitochondrial calcium uptake protein 1 (MICU1) regulates the uptake of mitochondrial calcium by the mitochondrial calcium uniporter (MCU) and subsequently controls aerobic metabolism and oxidative stress in mitochondria, but research considering oocytes remains unreported. We evaluated whether the addition of MICU1 modulators enhances mitochondrial activity, pyruvate metabolism, and developmental competence after warming of MII oocytes. METHODS: Retrieved MII oocytes of mice were classified as vitrified or control groups. After thawing, oocytes of vitrified group were cultured with or without DS16570511 (MICU1 inhibitor) and MCU-i4 (MICU1 activator) for 2 h. RESULTS: Mitochondrial Ca2+ concentration, pyruvate dephosphorylation level, and MICU1 expression of MII oocytes were significantly increased after vitrification. These phenomena were further exacerbated by the addition of MCU-i4 and reversed by the addition of DS16570511 after warming. However, the mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) in vitrified-warmed MII oocytes drop significantly after vitrification, which was improved after MCU-i4 treatment and decreased significantly after DS16570511 treatment. The vitrification process was able to elicit a development competence reduction. After parthenogenetic activation, incubation of the thawed oocytes with MCU-i4 did not alter the cleavage and blastocyst rates. Moreover, incubation of the thawed oocytes with DS16570511 reduced the cleavage and blastocyst rates. CONCLUSIONS: MICU1-mediated increasing mitochondrial calcium uptake after vitrification of the MII oocytes promoted the pyruvate oxidation, and this process may maintain oocyte development competence by compensating for the consumption of ATP under stress state.