ID3 regulates progesterone synthesis in bovine cumulus cells through modulation of mitochondrial function.

DNA binding inhibitory factor 3 (ID3) has been shown to have a key role in maintaining proliferation and differentiation. It has been suggested that ID3 may also affect mammalian ovarian function. However, the specific roles and mechanisms are unclear. In this study, the expression level of ID3 in cumulus cells (CCs) was inhibited by siRNA, and the downstream regulatory network of ID3 was uncovered by high-throughput sequencing. The effects of ID3 inhibition on mitochondrial function, progesterone synthesis, and oocyte maturation were further explored. The GO and KEGG analysis results showed that after ID3 inhibition, differentially expressed genes, including StAR, CYP11A1, and HSD3B1, were involved in cholesterol-related processes and progesterone-mediated oocyte maturation. Apoptosis in CC was increased, while the phosphorylation level of ERK1/2 was inhibited. During this process, mitochondrial dynamics and function were disrupted. In addition, the first polar body extrusion rate, ATP production and antioxidation capacity were reduced, which suggested that ID3 inhibition led to poor oocyte maturation and quality. The results will provide a new basis for understanding the biological roles of ID3 as well as cumulus cells.

Relativistic impulse approximation in the atomic ionization process induced by millicharged particles.

The millicharged particle has become an attractive topic to probe physics beyond the Standard Model. In direct detection experiments, the parameter space of millicharged particles can be constrained from the atomic ionization process. In this work, we develop the relativistic impulse approximation (RIA) approach, which can duel with atomic many-body effects effectively, in the atomic ionization process induced by millicharged particles. The formulation of RIA in the atomic ionization induced by millicharged particles is derived, and the numerical calculations are obtained and compared with those from free electron approximation and equivalent photon approximation. Concretely, the atomic ionizations induced by mllicharged dark matter particles and millicharged neutrinos in high-purity germanium (HPGe) and liquid xenon (LXe) detectors are carefully studied in this work. The differential cross sections, reaction event rates in HPGe and LXe detectors, and detecting sensitivities on dark matter particle and neutrino millicharge in next-generation HPGe and LXe based experiments are estimated and calculated to give a comprehensive study. Our results suggested that the next-generation experiments would improve 2-3 orders of magnitude on dark matter particle millicharge δ χ than the current best experimental bounds in direct detection experiments. Furthermore, the next-generation experiments would also improve 2-3 times on neutrino millicharge δ ν than the current experimental bounds.

Mutational spectrum of phenylketonuria in Jiangsu province.

UNLABELLED: Phenylketonuria (PKU) is caused by variants in the phenylalanine hydroxylase (PAH) gene. We systematically investigated all 13 exons of the PAH gene and their flanking introns in 31 unrelated patients and their parents using next-generation sequencing (NGS). A total of 33 different variants were identified in 58 of 62 mutant PAH alleles. The prevalent variants with a relative frequency of 5 % or more were c.721C > T, c.1068C > A, c.611A > G, c.1197A > T, c.728G > A, c.331C > T, and c.442-1G > A. One novel variant was identified in this study-c.699C > G. We studied genotype-phenotype correlations using the Guldberg arbitrary value (AV) system, which revealed a consistency rate of 38 % (8/21) among the 21 predicted phenotypes. The genotype-based prediction of BH4 responsiveness was also evaluated, and 14 patients (45.2 %) were predicted to be BH4 responsive. CONCLUSION: This study presents the spectrum of PAH variants in Jiangsu province. The information obtained from the genotype-based prediction of BH4 responsiveness might be used for the rational selection of candidates for BH4 testing. WHAT IS KNOWN: • Phenylketonuria (PKU) is caused by variants in the phenylalanine hydroxylase (PAH) gene. • The spectrum of PAH variants in different Chinese populations has been reported. What is new: • This is the first report on the spectrum of PAH variants in Jiangsu province. • This study identified one novel PAH variant-c.699C>G-and and tries to show a genotype-phenotype relationship also regarding BH4-responsiveness.

Hypoxia-mimetic agents inhibit proliferation and alter the morphology of human umbilical cord-derived mesenchymal stem cells.

BACKGROUND: The therapeutic efficacy of human mesenchymal stem cells (hMSCs) for the treatment of hypoxic-ischemic diseases is closely related to level of hypoxia in the damaged tissues. To elucidate the potential therapeutic applications and limitations of hMSCs derived from human umbilical cords, the effects of hypoxia on the morphology and proliferation of hMSCs were analyzed. RESULTS: After treatment with DFO and CoCl2, hMSCs were elongated, and adjacent cells were no longer in close contact. In addition, vacuole-like structures were observed within the cytoplasm; the rough endoplasmic reticulum expanded, and expanded ridges were observed in mitochondria. In addition, DFO and CoCl2 treatments for 48 h significantly inhibited hMSCs proliferation in a concentration-dependent manner (P < 0.05). This treatment also increased the number of cells in G0/G1 phase and decreased those in G2/S/M phase. CONCLUSIONS: The hypoxia-mimetic agents, DFO and CoCl2, alter umbilical cord-derived hMSCs morphology and inhibit their proliferation through influencing the cell cycle.

[Differential proteomic analysis in human umbilical cord mesenchymal stem cells induced by cobalt chloride].

OBJECTIVE: To analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry. METHODS: 2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions. RESULTS: 2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated. CONCLUSION: The effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.

Phosphoproteome profile of human lung cancer cell line A549.

As an in vitro model for type II human lung cancer, A549 cells resist cytotoxicity via phosphorylation of proteins as demonstrated by many studies. However, to date, no large-scale phosphoproteome investigation has been conducted on A549. Here, we performed a systematical analysis of the phosphoproteome of A549 by using mass spectrometry (MS)-based strategies. This investigation led to the identification of 337 phosphorylation sites on 181 phosphoproteins. Among them, 67 phosphoproteins and 230 phosphorylation sites identified appeared to be novel with no previous characterization in lung cancer. Based on their known functions as reported in the literature, these phosphoproteins were functionally organized into highly interconnected networks. Western blotting and immunohistochemistry analyses were performed to validate the expression of a bottleneck phosphoprotein YAP1 in cancer cell lines and tissues. This dataset provides a valuable resource for further studies on phosphorylation and lung carcinogenesis.

Quantitative phosphoproteomics of proteasome inhibition in multiple myeloma cells.

BACKGROUND: The proteasome inhibitor bortezomib represents an important advance in the treatment of multiple myeloma (MM). Bortezomib inhibits the activity of the 26S proteasome and induces cell death in a variety of tumor cells; however, the mechanism of cytotoxicity is not well understood. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the differential phosphoproteome upon proteasome inhibition by using stable isotope labeling by amino acids in cell culture (SILAC) in combination with phosphoprotein enrichment and LC-MS/MS analysis. In total 233 phosphoproteins were identified and 72 phosphoproteins showed a 1.5-fold or greater change upon bortezomib treatment. The phosphoproteins with expression alterations encompass all major protein classes, including a large number of nucleic acid binding proteins. Site-specific phosphopeptide quantitation revealed that Ser38 phosphorylation on stathmin increased upon bortezomib treatment, suggesting new mechanisms associated to bortezomib-induced apoptosis in MM cells. Further studies demonstrated that stathmin phosphorylation profile was modified in response to bortezomib treatment and the regulation of stathmin by phosphorylation at specific Ser/Thr residues participated in the cellular response induced by bortezomib. CONCLUSIONS/SIGNIFICANCE: Our systematic profiling of phosphorylation changes in response to bortezomib treatment not only advanced the global mechanistic understanding of the action of bortezomib on myeloma cells but also identified previously uncharacterized signaling proteins in myeloma cells.

[Comparative study on quality of Tongrentang red ginseng and Korean red ginseng--determination of ginsenosides and polysaccharides].

OBJECTIVE: To establish methods for quantitative determination of ginseng saponins, ginsenoside Rg1, Re, Rb1 and polysaccarides and compare the qualities of Tongrentang Red Ginseng and Korean Red Ginseng. METHOD: Macroreticular resin-colorimetric method was developed to determine ginseng saponins and a new HPLC method with gradient eluents was established for determination of ginsenoside Rg1, Re, Rb1. For ginseng polysaccharides, phenol-oil of vitriol colorimetric method was developed and some factors were also optimized. RESULT: The content of ginseng saponins in Tongrentang Red Ginseng was not lower than that of Korean Red Ginseng. Ginsenoside Rg1 and Rb1 in Tongrentang Red Ginseng were higher than those in Korean Red Ginseng, while Ginsenoside Re was slightly lower than that of Korean Red Ginseng. However, the amount of Ginseng Polysaccharides in Tongrentang Red Ginseng was greater than those in Korean Red Ginseng. CONCLUSION: The contents of ginseng saponins and ginsenoside Rg1, Re, Rb1 in Tongrentang Red Ginseng were not lower than that in Korean Red Ginseng. The methods for determination of ginsenosides and ginseng polysaccharides were quite accurate and reliable to the quality control of Ginseng.