Integrated lipid metabolomics and proteomics analysis reveal the pathogenesis of polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrinological and metabolic disorder that can lead to female infertility. Lipid metabolomics and proteomics are the new disciplines in systems biology aimed to discover metabolic pathway changes in diseases and diagnosis of biomarkers. This study aims to reveal the features of PCOS to explore its pathogenesis at the protein and metabolic level. METHODS: We collected follicular fluid samples and granulosa cells of women with PCOS and normal women who underwent in vitro fertilization(IVF) and embryo transfer were recruited. The samples were for the lipidomic study and the proteomic study based on the latest metabolomics and proteomics research platform. RESULTS: Lipid metabolomic analysis revealed abnormal metabolism of glycerides, glycerophospholipids, and sphingomyelin in the FF of PCOS. Differential lipids were strongly linked with the rate of high-quality embryos. In total, 144 differentially expressed proteins were screened in ovarian granulosa cells in women with PCOS compared to controls. Go functional enrichment analysis showed that differential proteins were associated with blood coagulation and lead to follicular development disorders. CONCLUSION: The results showed that the differential lipid metabolites and proteins in PCOS were closely related to follicle quality,which can be potential biomarkers for oocyte maturation and ART outcomes.

Differential expression of follicular fluid exosomal microRNA in women with diminished ovarian reserve.

PURPOSE: Decreased ovarian reserve function is mainly characterized by female endocrine disorders and fertility decline. Follicular fluid (FF) exosomal microRNAs (miRNAs) have been shown to regulate the function of granulosa cells (GCs). The present study explored differentially expressed miRNAs (DEmiRNAs) in patients with diminished ovarian reserve (DOR). METHODS: FF was collected from 12 DOR patients and 12 healthy controls. DEmiRNAs between the two groups were identified and analyzed using high-throughput sequencing technology and validated by real-time quantitative PCR (RT-qPCR). RESULTS: A total of 592 DEmiRNAs were identified using high-throughput miRNA sequencing, of which 213 were significantly upregulated and 379 were significantly downregulated. The sequencing results were further validated by RT-qPCR. These DEmiRNA target genes were mainly involved in the cancer pathway, phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, regulation of actin cytoskeleton signaling pathway, and biological processes related to protein binding, nucleoplasm, cytoplasm, and cell membrane. CONCLUSION: FF exosomal miRNAs are significantly differentially expressed in DOR patients versus non-DOR patients, underscoring their crucial role in regulating the pathogenesis of DOR.

Effects of Cu (II) on the Growth of Chlorella vulgaris and Its Removal Efficiency of Pollutants in Synthetic Piggery Digestate.

C. vulgaris has a positive effect on the removal of nutrients from pig farm biogas slurry. However, swine wastewater often contains heavy metal ions, such as Cu (II), which may have impacts on the nutrient removal performance of C. vulgaris. Additionally, the heavy metal ions in wastewater can be adsorbed by microalgae. In this study, the stress effect of Cu (II) on the growth of Chlorella vulgaris, the Cu (II) removal by microalgae, and the effect of different concentrations of Cu (II) on the nutrient removal efficiency of C. vulgaris in biogas slurries were explored. The results showed that the microalgae biomass of microalgae on the sixth day of the experiment was the highest in the treatment with a Cu (II) concentration of 0.5 mg/L, which was 30.1% higher than that of the 2.5 mg/L group. C. vulgaris had higher removal efficiencies of Cu (II) at a Cu (II) concentration of 0.1~1.5 mg/L. The-OH, C=O, -COOH, and C-O groups on the surface of the algal cells play a significant role in the removal of Cu (II). The removal rates of COD, NH3-N, TN, and TP by C. vulgaris at a Cu (II) concentration of 0.5 mg/L were the highest, which were 89.0%, 53.7%, 69.6%, and 47.3%, respectively.

Unraveling the Electronic Structure and Dynamics of the Atomically Dispersed Iron Sites in Electrochemical CO2 Reduction.

Single-atom catalysts with a well-defined metal center open unique opportunities for exploring the catalytically active site and reaction mechanism of chemical reactions. However, understanding of the electronic and structural dynamics of single-atom catalytic centers under reaction conditions is still limited due to the challenge of combining operando techniques that are sensitive to such sites and model single-atom systems. Herein, supported by state-of-the-art operando techniques, we provide an in-depth study of the dynamic structural and electronic evolution during the electrochemical CO2 reduction reaction (CO2RR) of a model catalyst comprising iron only as a high-spin (HS) Fe(III)N4 center in its resting state. Operando 57Fe Mössbauer and X-ray absorption spectroscopies clearly evidence the change from a HS Fe(III)N4 to a HS Fe(II)N4 center with decreasing potential, CO2- or Ar-saturation of the electrolyte, leading to different adsorbates and stability of the HS Fe(II)N4 center. With operando Raman spectroscopy and cyclic voltammetry, we identify that the phthalocyanine (Pc) ligand coordinating the iron cation center undergoes a redox process from Fe(II)Pc to Fe(II)Pc-. Altogether, the HS Fe(II)Pc- species is identified as the catalytic intermediate for CO2RR. Furthermore, theoretical calculations reveal that the electroreduction of the Pc ligand modifies the d-band center of the in situ generated HS Fe(II)Pc- species, resulting in an optimal binding strength to CO2 and thus boosting the catalytic performance of CO2RR. This work provides both experimental and theoretical evidence toward the electronic structural and dynamics of reactive sites in single-Fe-atom materials and shall guide the design of novel efficient catalysts for CO2RR.

Pollutant removal from swine wastewater and kinetics in constructed rapid infiltration system (CRI system).

The purpose of this paper is centred on the kinetics of removal of main pollutants in wastewater and to compared different hydraulic loading conditions of the constructed rapid infiltration system (CRI system) in terms of removal efficiencies, effluent concentrations, mass removal rate (MRR), and the first-order removal rate coefficient (k) of COD, TOC, NH4+-N, TN, and TP. The results showed that the higher the hydraulic loading, the higher the effluent concentration. The results that synthesized hydraulic loading, effluent concentrations, removal efficiencies, and other conditions showed that the best hydraulic loading was 40 cm/d. When the hydraulic load was 40 cm/d, the effluent average concentrations of COD, TOC, NH4+-N, TN, TP, Cu2+ and the removal efficiencies were 27.31 ± 16.40 mg/L, 86.11%, 10.55 ± 5.25 mg/L, 84.64%, 0.59 ± 0.87 mg/L, 99.60%, 143.31 ± 14.77 mg/L, 7.04%, 5.64 ± 1.38 mg/L, 79.20%, and 0.13 ± 0.47 mg/L, 97.51%, respectively. According to a kinetic study of the primary pollutants, the MRR increased with an increase in the hydraulic loading, except for ammonia nitrogen. CRI-3, CRI-4 were high significant correlated with ammonia nitrogen (with R2 = 93.65% and R2 = 95.03%, respectively), while CRI-2, CRI-3, and CRI-4 were high significant correlated with total nitrogen (with R2 = 94.56%, R2 = 96.70% and R2 = 96.56% respectively).

Degradation of ammonia gas by Cu2O/{001}TiO2 and its mechanistic analysis.

A heterogeneous composite catalyst Cu2O/{001}TiO2 was successfully prepared by the impregnation-reduction method. With ammonia as the target pollutant, the degradation performance and degradation mechanism analysis of the prepared composite catalyst were investigated, providing technology for the application of photocatalysis technology in ammonia treatment reference. The catalysts were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), specific surface area (BET), fluorescence spectrum (PL) and UV-visible absorption (UV-Vis). The results showed: compared with single {001}TiO2, the addition of Cu2O to form a composite catalyst can reduce the recombination of electron-hole pairs, resulting in increased absorption intensity in the visible light range, decreased band gap width, and finally improved the degradation performance. When the composite ratio is 1 : 10, the specific surface area is the largest, which is 72.51 m2 g-1, and the degradation rate of ammonia is also the highest maintained at 85%. After repeated use for 5 times, the degradation rate of ammonia decreases gradually due to the loss of catalyst and photo-corrosion. In the whole reaction process, surface adsorbed water and associated hydroxyl radical participate in the ammonia degradation reaction, and finally form free hydroxyl radical and NO3 -. It provides some theoretical support for ammonia gas treatment, which is of great significance to protect the environment.

Preparation of F-doped H2Ti3O7-{104} nanorods with oxygen vacancies using TiOF2 as precursor and its photocatalytic degradation activity.

Photocatalytic degradation is an eco-friendly and sustainable method for the treatment of water pollutants especially tetracycline hydrochloride (TCH). Herein, we developed F-doped H2Ti3O7-{104} nanorods with oxygen vacancies using TiOF2 as a precursor by simple alkali hydrothermal and ion-exchange methods. The phase structure, surface composition, optical properties, specific surface areas and charge separation were analysed by a series of measurements. The effects of KOH concentration on the structure and properties of H2Ti3O7 were investigated. It is confirmed that the TiOF2/H2Ti3O7 composite can be formed in low concentration KOH solution (1 mol L-1), while the H2Ti3O7 single phase can be formed in high concentration KOH solution (>3 mol L-1). The prepared F-doped H2Ti3O7-{104} nanorods provide a high specific surface area of 457 m2 g-1 and a macroporous volume of 0.69 cm3 g-1. The appropriate mesoporous structure of the photocatalyst makes TCH have a stronger affinity on its surface, which is more conducive to the subsequent photodegradation. Moreover, a synergistic mechanism of photosensitization and ligand-metal charge transfer (LMCT) in the photocatalytic degradation of TCH was proposed. In addition, the prepared F-doped H2Ti3O7-{104} nanorods showed excellent cycle stability and resistance to light corrosion. After five cycles of photodegradation, the degradation rate of TCH was only reduced from 92% to 83%. This low-cost strategy could be used for the mass production of efficient photocatalysts, which can be used for TCH clean-up in wastewater treatment.

Transformation of novel TiOF2 nanoparticles to cluster TiO2-{001/101} and its degradation of tetracycline hydrochloride under simulated sunlight.

The anatase type cluster TiO2-{001/101} was rapidly generated by a one-step hydrothermal method. The transformation process of coral-like TiOF2 nanoparticles to cluster TiO2-{001/101} was investigated for the first time, and the sensitization between cluster TiO2-{001/101} and tetracycline hydrochloride (TCH) was also discussed. The degradation rate of TCH by cluster TiO2-{001/101} under simulated sunlight was 92.3%, and the total removal rate was 1.76 times that of P25. Besides, cluster TiO2-{001/101} settles more easily than P25 in deionized water. The study showed that cluster TiO2-{001/101} derived from coral-like TiOF2 nanoparticles had a strong adsorption effect on TCH, which was attributed to the oxygen vacancy (Ov) and {001} facets of cluster TiO2-{001/101}. The strong adsorption effect promoted the sensitization between cluster TiO2-{001/101} and TCH, and widened the visible light absorption range of cluster TiO2-{001/101}. In addition, the fluorescence emission spectrum showed that cluster TiO2-{001/101} had a lower luminous intensity, which was attributed to the heterojunction formed by {001} facets and {101} facets that reduces the recombination rate of carriers. It should be noted that cluster TiO2-{001/101} still has good degradation performance for TCH after five cycles of degradation. This study provides a new idea for the synthesis of cluster TiO2-{001/101} with high photocatalytic performance for the treatment of TCH wastewater.

Homocysteine impairs porcine oocyte quality via deregulation of one-carbon metabolism and hypermethylation of mitochondrial DNA†.

Homocysteine (Hcy) is an intermediate in the one-carbon metabolism that donates methyl groups for methylation processes involved in epigenetic gene regulation. Although poor oocyte quality in polycystic ovarian syndrome (PCOS) patients is associated with elevated Hcy concentration in serum and follicular fluid, whether Hcy directly affects oocyte quality and its mechanisms are poorly understood. Here we show that Hcy treatment impaired oocyte quality and developmental competence, indicated by significantly reduced survival rate, polar body extrusion rate, and cleavage rate. Hcy treatment resulted in mitochondrial dysfunction, with increased production of mitochondrial ROS, reduced mtDNA copy number, and the expression of 7 out of 13 mtDNA-encoded genes and 2 ribosome RNA genes, 12S rRNA and 16S rRNA. Upon Hcy treatment, the expression of one-carbon metabolic enzymes and DNMT1 was enhanced. Interestingly, DNA methyltransferase inhibitor 5'AZA rescued Hcy-induced mitochondrial dysfunction, impaired oocyte quality and developmental competence. Concurrently, expression of one-carbon metabolic enzymes and methylation status of mtDNA coding sequences were also normalized, at least partially, by 5'AZA treatment. Our findings not only extend the understanding about how Hcy induces poor oocyte quality, but also contribute to a novel angle of identifying targets for enhancing the quality of oocyte from PCOS patients.

The dynamic pattern of PLIN3 in pig oocytes and cumulus cells during in vitro maturation.

Lipid droplets (LDs) are the main energy resource for porcine preimplantation embryonic development. PLIN3 has been implicated in LD formation and regulation. Therefore, this study aimed to detect the dynamic pattern of PLIN3 in pig oocytes and cumulus cells (CC) during in vitro maturation (IVM), and to determine the relationship between PLIN3 and LD content. IVM with cumulus-enclosed oocytes (CEO), cumulus-denuded oocytes (DO) and the CCs denuded from the corresponding oocytes (DCC) was performed in porcine follicular fluid (PFF) or PFF-free optimized medium. DO and the DCC were cultured together under the same conditions as described above, while the DO was named DTO and the DCC was named DTCC in this group. Firstly, our results revealed LDs distributed widely in oocytes and CC, while the PLIN3 protein coated these LDs and spread out ubiquitously in the cytoplasm. Secondly, not only the mRNA level but also at protein level of PLIN3 in immature naked oocytes (IO) was higher than that in matured CEO, DO and DTO. Although PLIN3 was expressed at lower levels in CC from immature oocytes (ICC), the protein level of PLIN3 was comparably higher in the ECC and DCC groups. The triglyceride (TG) content in CEO and DO was significantly less abundant compared with that in IO. Therefore, our results indicated that co-culturing of oocytes and CC might affect PLIN3 expression levels in CC but not in oocytes. Lipid accumulation in pig oocytes during maturation might be affected by PLIN3 cross-talk between oocytes and CC.

LC3-Dependent Autophagy in Pig 2-Cell Cloned Embryos Could Influence the Degradation of Maternal mRNA and the Regulation of Epigenetic Modification.

In this study, the distribution as well as the effect of autophagy on reprogramming in pig cloned embryos were observed immediately after somatic cell nuclear transfer. Results showed that the LC3 was at the highest level in cloned embryos at 2-cell stage, and it decreased with the development from 2-cell stage to blastocyst. Different to cloned embryos, the intensity of LC3 in parthenogenetic activation (PA) embryos was at the highest level at 4-cell stage. A markedly higher level of Bmp15, H1foo, and Dppa3 was shown in cloned embryos at 2-cell stage (p < 0.05 or p < 0.01), but a significantly lower level of LC3, Sox2, and eIF1A was observed at 4-cell stage (p < 0.05), compared with PA embryos. When the efficient interfering by the LC3 siRNA was performed on the cloned embryos (p < 0.01), not only the mRNA level of maternal Cyclin B, Bmp15, Gdf9, c-mos, H1foo, and Dppa3 was increased significantly (p < 0.05), but also the expression of Dnmt1 and Dnmt3b was obviously upregulated (p < 0.05). Although the expression of Sox2 and Oct4 is not changed, the expression of Stat3 decreased significantly (p < 0.05). Furthermore with the treatment of 200 nM rapamycin, the expression of eIF1A and Stat3 was significantly increased at 4-cell stage. In conclusion, the LC3-dependent autophagy mainly occurred in cloned embryos at 2-cell stage, but at 4-cell stage in PA embryos. In addition, the modulation of autophagy could affect genome activation by influencing the degradation of maternal mRNA and regulating the expression of DNA methyltransferase.

Dynamic Reorganization of Nucleosome Positioning in Somatic Cells after Transfer into Porcine Enucleated Oocytes.

The nucleosome, the fundamental structural unit of chromatin, is a critical regulator of gene expression. The mechanisms governing changes to nucleosome occupancy and positioning during somatic cell reprogramming remain poorly understood. We established a method for generating genome-wide nucleosome maps of porcine embryonic fibroblasts (PEF), reconstructed 1-cell embryos generated by somatic cell nuclear transfer (SCNT), and fertilized zygotes (FZ) using MNase sequencing with only 1,000 cells. We found that donor PEF chromatin, especially X chromosome, became more open after transfer into porcine oocytes and nucleosome occupancy decreased in promoters but increased in the genic regions. Nucleosome arrangements around transcriptional start sites of genes with different expression levels in somatic cells tended to become transcriptionally silent in SCNT; however, some pluripotency genes adopted transcriptionally active nucleosome arrangements. FZ and SCNT had similar characteristics, unlike PEF. This study reveals the dynamics and importance of nucleosome positioning and chromatin organization early after reprogramming.

Genome-scale identification of nucleosome organization by using 1000 porcine oocytes at different developmental stages.

The nucleosome is the basic structural unit of chromosomes, and its occupancy and distribution in promoters are crucial for the regulation of gene expression. During the growth process of porcine oocytes, the "growing" oocytes (SF) have a much higher transcriptional activity than the "fully grown" oocytes (BF). However, the chromosome status of the two kinds of oocytes remains poorly understood. In this study, we profiled the nucleosome distributions of SF and BF with as few as 1000 oocytes. By comparing the altered regions, we found that SF tended toward nucleosome loss and more open chromosome architecture than BF did. BF had decreased nucleosome occupancy in the coding region and increased nucleosome occupancy in the promoter compared to SF. The nucleosome occupancy of SF was higher than that of BF in the GC-poor regions, but lower than that of BF in the GC-rich regions. The nucleosome distribution around the transcriptional start site (TSS) of all the genes of the two samples was basically the same, but the nucleosome occupancy around the TSS of SF was lower than that of BF. GO functional annotation of genes with different nucleosome occupancy in promoter showed the genes were mainly involved in cell, cellular process, and metabolic process biological process. The results of this study revealed the dynamic reorganization of porcine oocytes in different developmental stages and the critical role of nucleosome arrangement during the oocyte growth process.

Effects of combined exposure to 17α-ethynylestradiol and dibutyl phthalate on the growth and reproduction of adult male zebrafish (Danio rerio).

To evaluate the combined effects of 17α-ethynylestradiol (EE2) and dibutyl phthalate (DBP) on the growth and reproduction of male zebrafish, three-month-old fish were exposed to 0.005 or 0.020µg/L EE2, 100 or 500µg/L DBP or their binary mixtures under semi-static conditions. Investigated parameters include the length, weight, condition factor, vitellogenin (VTG) induction, acyl-CoA oxidase (AOX) protein level, histopathological alteration of testis, liver and gill, and reproductive capacity. After 21d exposure, no statistical difference was found among the weights, lengths and condition factors of different treatment groups. In all binary mixture groups, decreased VTG levels were detected compared to EE2-only groups; and the AOX levels were significantly lower than DBP-only treatments while both chemicals can individually induce AOX synthesis. Therefore, EE2 and DBP may act additively on VTG and antagonistically on AOX induction in males. After 45d exposure, delayed gametogenesis was observed for the DBP-only groups, indicated by fewer spermatozoa and more spermatocytes, which was further aggravated with the addition of EE2. The developmental delay of testis partially recovered after a 30d depuration in clean water. Combined exposure also caused liver and gill lesions, which were not alleviated during the 30d depuration, suggesting a nonreversible harmful effect the same as single exposure. Mixed EE2 and DBP were observed to impair the reproductive capability (the fecundity and fertilization rate) of males, while single exposure did not. Co-exposed to 0.020µg/L EE2 and 100µg/L DBP promoted the early hatching of offspring (F1 generation) at 48h post-fertilization (hpf), but the survival rates of the F1 generation were similar in all treatments. Our findings indicate that the effects of mixed EE2 and DBP at environmentally relevant levels can be either antagonistic or additive relying on the specific toxicological endpoints and the respective doses of each chemical.

Bio-electro-Fenton system for enhanced estrogens degradation.

The feasibility of removing estrogens including 17ß-estradiol (E2) and 17α-ethynyl-estradiol (EE2) was studied in a bio-electro-Fenton (BEF) system equipped with a Fe@Fe2O3/non-catalyzed carbon felt (NCF) composite cathode. E2 and EE2 were removed by reactive oxidants, produced by bio-electro-Fenton system and zero-valent iron/O2 system, as well as adsorption. Under closed-circuit condition, 81% of E2 and 56% of EE2 were removed within 10h in the system, in which the highest concentration of total iron ions and H2O2 reached 81 and 1.2mg/L, respectively. The maximum power density of BEF system equipped with Fe@Fe2O3/NCF electrode was 4.35 W/m(3). Two intermediates of E1 and 6-OH-E2 were identified during Fenton oxidation of E2. This study demonstrates the degradation fate of E2 and EE2 in a BEF system equipped with Fe@Fe2O3/NCF electrodes, which provides a promising and cost-effective solution for the removal of recalcitrant contaminants with simultaneous power generation.