Cell-Free Seminal mRNA and MicroRNA Exist in Different Forms

BACKGROUND: The great interest in cell-free mRNA, microRNA (miRNA) as molecular biomarkers for clinical applications, and as 'signaling' molecules for intercellular communication highlights the need to reveal their physical nature. Here this issue was explored in human cell-free seminal mRNA (cfs-mRNA) and miRNA (cfs-miRNA). METHODOLOGY/PRINCIPAL FINDINGS: Selected male reproductive organ-specific mRNAs, miRNAs, and piRNAs were quantified by quantitative real-time PCR in all experiments. While the stability of cfs-miRNA assessed by time-course analysis (up to 24 h at room temperature) was similar with cfs-mRNA, the reductive changes between cfs-miRNA and cfs-mRNA after filtration and Triton X-100 treatment on seminal plasma were very different, implying their different physical nature. Seminal microvesicles (SMVs) were then recovered and proportions of cfs-mRNA and cfs-miRNA within SMVs were quantified. The amounts of SMVs- sequestered cfs-mRNAs almost were the same as total cfs-mRNA, and were highly variable depending on the different sizes of SMVs. But most of cfs-miRNA was independent of SMVs and existed in the supernatant. The possible form of cfs-miRNA in the supernatant was further explored by filtration and protease K digestion. It passed through the 0.10-µm pore, but was degraded dramatically after intense protease K digestion. CONCLUSIONS/SIGNIFICANCE: The predominant cfs-mRNA is contained in SMVs, while most cfs-miRNA is bound with protein complexes. Our data explained the stability of extracellular RNAs in human semen, and shed light on their origins and potential functions in male reproduction, and strategy of developing them as biomarkers of male reproductive system

P-CSREC: A New Approach for Personalized Cloud Service Recommendation

It is becoming a challenging issue for users to choose a satisfied service to fit their need due to the rapid growing number of cloud services and the vast amount of service type varieties. This paper proposes an effective cloud service recommendation approach, named personalized cloud service recommendation (P-CSREC), based on the characterization of heterogeneous information network, the use of association rule mining, and the modeling and clustering of user interests. First, a similarity measure is defined to improve the average similarity (AvgSim) measure by the inclusion of the subjective evaluation of users’ interests. Based on the improved AvgSim, a new model for measuring the user interest is established. Second, the traditional K-Harmonic Means (KHM) clustering algorithm is improved by means of involving multi meta-paths to avoid the convergence of local optimum. Then, a frequent pattern growth (FP-Growth) association rules algorithm is proposed to address the issue and the limitation of traditional association rule algorithms to offer personalization in recommendation. A new method to define a support value of nodes is developed using the weight of user’s score. In addition, a multi-level FP-Tree is defined based on the multi-level association rules theory to extract the relationship in higher level. Finally, a combined user interest with the improved KHM clustering algorithm and the improved FP-Growth algorithm is provided to improve accuracy of cloud services recommendation to target users. The experimental results demonstrated the effectiveness of the proposed approach in improving the computational efficiency and recommendation accuracy

Cornel Iridoid Glycoside Inhibits Tau Hyperphosphorylation via Regulating Cross-Talk Between GSK-3β and PP2A Signaling

Neurofibrillary pathology contributes to neuronal dysfunction and correlates with the clinical progression of Alzheimer’s disease (AD). Tau phosphorylation is mainly regulated by a balance of glycogen synthase kinase-3β (GSK-3β) and protein phosphatase 2A (PP2A) activities. Cornel iridoid glycoside (CIG) is a main component extracted from Cornus officinalis. The purpose of this study was to investigate the effects of CIG on GSK-3β and PP2A, thus to explore the mechanisms of CIG to inhibit tau hyperphosphorylation. The rat model of tau hyperphosphorylation was established by intraventricular injection of wortmannin and GF-109203X (GFX) to activate GSK-3β. The results showed that intragastrical administration of CIG inhibited tau hyperphosphorylation in the brain of rats induced by wortmannin/GFX. The results in vivo and in vitro exhibited that CIG inhibited tau hyperphosphorylation and GSK-3β over-activation. In the mechanism of action, CIG’s attenuating GSK-3β activity was found to be dependent on PI3K/AKT signaling pathway. PP2A catalytic C subunit (PP2Ac) siRNA abrogated the effect of CIG on PI3K/AKT/GSK-3β. Additionally and crucially, we also found that CIG inhibited the demethylation of PP2Ac at Leu309 in vivo and in vitro. It enhanced PP2A activity, decreased tau hyperphosphorylation, and protected cell morphology in okadaic acid (OA)-induced cell model in vitro. PP2Ac siRNA abated the inhibitory effect of CIG on tau hyperphosphorylation. Moreover, CIG inhibited protein phosphatase methylesterase-1 (PME-1) and demethylation of PP2Ac, enhanced PP2A activity, and decreased tau hyperphosphorylation in PME-1-transfectd cells. Taken together, CIG inhibited GSK-3β activity via promoting P13K/AKT and PP2A signaling pathways. In addition, CIG also elevated PP2A activity via inhibiting PME-1-induced PP2Ac demethylation to inhibit GSK-3β activity, thus regulated the cross-talk between GSK-3β and PP2A signaling and consequently inhibited tau hyperphosphorylation. These results suggest that CIG may be a promising agent for AD therapy

Genetic changes in fetal cerebral cortex after maternal exposure to sevoflurane

Purpose: To investigate the acute changes in transcriptome of radial glial progenitor cells after maternal exposure to sevoflurane.Methods: Two groups of sample data were collected from the data set in the GEO database. Pregnant mice in sevoflurane group were exposed to 2.5 % sevoflurane for 6 h on day 14.5 of the pregnancy, while the mice in control group were exposed to 100 % oxygen for 6 h. At the end of the exposure period, the cerebral cortex of the two groups of fetuses was isolated and analyzed by RNA sequencing. Differentially expressed genes (DEG) were analyzed using limma package of R language. The Gene Ontology (GO) and KEGG pathway enrichment were based on DEG through a cluster profile package of R. Moreover, protein-protein interactions (PPI) network construction and central gene prediction were carried out using a string database and R package.Results: Bioinformatics analysis revealed 289 up-regulated genes and 311 down-regulated DEGs, respectively. Gene Ontology and KEGG enrichment analysis revealed terms related to neural development and transcriptional function. Based on the central genes of the PPI network, it was found that certain genes may play significant roles in the regulation of neural development. These genes are hnRNPM, AURKA, NCBP, SRSF6, ASF1B, HNRNPA2B1, DDX21, H3F3B, KPNA2 and ABCE1 (p < 0.05).Conclusion: The findings of this study suggest that hub genes and a variety of signal pathways may play key roles in the development of radial glial progenitor cells

Identification and characterization of the expression profile of microRNAs in Anopheles anthropophagus

BACKGROUND: Anopheles anthropophagus, one of the most important mosquito-borne disease vectors in Asia, mainly takes blood meals from humans and transmits both malaria and filariae. MicroRNAs (miRNAs) are small non-coding RNAs, and play a critical role in many cellular processes, including development, differentiation, apoptosis and innate immunity. METHODS: We investigated the global miRNA expression profile of male and female adults of A. anthropophagus using illumina Hiseq2000 sequencing combined with Northern blot. RESULTS: By using the miRNAs of the closely-related species Anopheles gambiae and Aedes aegypti as reference, we obtained 102 miRNAs candidates out of 12.43 million raw sequencing reads for male and 16.51 million reads for female, with 81 of them found as known miRNAs in An. gambiae and/or Ae. aegypti, and the remaining 21 miRNAs were considered as novel. By analyzing the revised read count of miRNAs in male and female, 29 known miRNAs show sexual difference expression: >2-fold in the read count of the same miRNAs in male and female. Especially for miR-989, which is highly expressed in the female mosquitoes, but shows almost no detected expression in male mosquitoes, indicating that miR-989 may be involved in the physiological activity of female mosquito adults. The expression of four miRNAs in different growth stages of mosquito were further identified by Northern blot. Several miRNAs show the stage-specific expression, of which miR-2943 only expressed in the egg stage, suggesting that miR-2943 may be associated with the development of mosquito eggs. CONCLUSIONS: The present study represents the first global characterization of An. anthropophagus miRNAs in sexual differences and stage-specific functions. A better understanding of the functions of these miRNAs will offer new insights in mosquito biology and has implications for the effective control of mosquito-borne infectious diseases

Musical experience may help the brain respond to second language reading

A person's native language background exerts constraints on the brain's automatic responses while learning a second language. It remains unclear, however, whether and how musical experience may help the brain overcome such constraints and meet the requirements of a second language. This study compared native Chinese English learners who were musicians, non-musicians and native English readers on their automatic brain automatic integration of English letter-sounds with an ERP cross-modal audiovisual mismatch negativity paradigm. The results showed that native Chinese-speaking musicians successfully integrated English letters and sounds, but their non-musician peers did not, despite of their comparable English learning experience and proficiency level. However, native Chinese-speaking musicians demonstrated enhanced cross-modal MMN for both synchronized and delayed letter-sound integration, while native English readers only showed enhanced cross-modal MMN for synchronized integration. Moreover, native Chinese-speaking musicians showed stronger theta oscillations when integrating English letters and sounds, suggesting that they had better top-down modulation. In contrast, native English readers showed stronger delta oscillations for synchronized integration, and their cross-modal delta oscillations significantly correlated with English reading performance. These findings suggest that long-term professional musical experience may enhance the top-down modulation, then help the brain efficiently integrating letter-sounds required by the second language. Such benefits from musical experience may be different from those from specific language experience in shaping the brain's automatic responses to reading.Peer reviewe