13 research outputs found

    RNA pull-down-confocal nanoscanning (RP-CONA), a novel method for studying RNA/protein interactions in cell extracts that detected potential drugs for Parkinson’s disease targeting RNA/HuR complexes

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    MicroRNAs (miRNAs, miRs) are a class of small non-coding RNAs that regulate gene expression through specific base-pair targeting. The functional mature miRNAs usually undergo a two-step cleavage from primary miRNAs (pri-miRs), then precursor miRNAs (pre-miRs). The biogenesis of miRNAs is tightly controlled by different RNA-binding proteins (RBPs). The dysregulation of miRNAs is closely related to a plethora of diseases. Targeting miRNA biogenesis is becoming a promising therapeutic strategy. HuR and MSI2 are both RBPs. MiR-7 is post-transcriptionally inhibited by the HuR/MSI2 complex, through a direct interaction between HuR and the conserved terminal loop (CTL) of pri-miR-7-1. Small molecules dissociating pri-miR-7/HuR interaction may induce miR-7 production. Importantly, the miR-7 levels are negatively correlated with Parkinson’s disease (PD). PD is a common, incurable neurodegenerative disease causing serious motor deficits. A hallmark of PD is the presence of Lewy bodies in the human brain, which are inclusion bodies mainly composed of an aberrantly aggregated protein named α-synuclein (α-syn). Decreasing α-syn levels or preventing α-syn aggregation are under investigation as PD treatments. Notably, α-syn is negatively regulated by several miRNAs, including miR-7, miR-153, miR-133b and others. One hypothesis is that elevating these miRNA levels can inhibit α-syn expression and ameliorate PD pathologies. In this project, we identified miR-7 as the most effective α-syn inhibitor, among the miRNAs that are downregulated in PD, and with α-syn targeting potentials. We also observed potential post-transcriptional inhibition on miR-153 biogenesis in neuroblastoma, which may help to uncover novel therapeutic targets towards PD. To identify miR-7 inducers that benefit PD treatment by repressing α-syn expression, we developed a novel technique RNA Pull-down Confocal Nanoscaning (RP-CONA) to monitor the binding events between pri-miR-7 and HuR. By attaching FITC-pri-miR-7-1-CTL-biotin to streptavidin-coated agarose beads and incubating them in human cultured cell lysates containing overexpressed mCherry-HuR, the bound RNA and protein can be visualised as quantifiable fluorescent rings in corresponding channels in a confocal high-content image system. A pri-miR-7/HuR inhibitor can decrease the relative mCherry/FITC intensity ratio in RP-CONA. With this technique, we performed several small-scale screenings and identified that a bioflavonoid, quercetin can largely dissociate the pri-miR-7/HuR interaction. Further studies proved that quercetin was an effective miR-7 inducer as well as α-syn inhibitor in HeLa cells. To understand the mechanism of quercetin mediated α-syn inhibition, we tested the effects of quercetin treatment with miR-7-1 and HuR knockout HeLa cells. We found that HuR was essential in this pathway, while miR-7 hardly contributed to the α-syn inhibition. HuR can directly bind an AU-rich element (ARE) at the 3’ untranslated region (3’-UTR) of α-syn mRNA and promote translation. We believe quercetin mainly disrupts the ARE/HuR interaction and disables the HuR-induced α-syn expression. In conclusion, we developed and optimised RP-CONA, an on-bead, lysate-based technique detecting RNA/protein interactions, as well as identifying RNA/protein modulators. With RP-CONA, we found quercetin inducing miR-7 biogenesis, and inhibiting α-syn expression. With these beneficial effects, quercetin has great potential to be applied in the clinic of PD treatment. Finally, RP-CONA can be used in many other RNA/protein interactions studies

    RNA-targeted Therapies and High-throughput Screening Methods

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    RNA-binding proteins (RBPs) are involved in regulating all aspects of RNA metabolism, including processing, transport, translation, and degradation. Dysregulation of RNA metabolism is linked to a plethora of diseases, such as cancer, neurodegenerative diseases, and neuromuscular disorders. Recent years have seen a dramatic shift in the knowledge base, with RNA increasingly being recognised as an attractive target for precision medicine therapies. In this article, we are going to review current RNA-targeted therapies. Furthermore, we will scrutinise a range of drug discoveries targeting protein-RNA interactions. In particular, we will focus on the interplay between Lin28 and let-7, splicing regulatory proteins and survival motor neuron (SMN) pre-mRNA, as well as HuR, Musashi, proteins and their RNA targets. We will highlight the mechanisms RBPs utilise to modulate RNA metabolism and discuss current high-throughput screening strategies. This review provides evidence that we are entering a new era of RNA-targeted medicine

    Rec4Ad: A Free Lunch to Mitigate Sample Selection Bias for Ads CTR Prediction in Taobao

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    Click-Through Rate (CTR) prediction serves as a fundamental component in online advertising. A common practice is to train a CTR model on advertisement (ad) impressions with user feedback. Since ad impressions are purposely selected by the model itself, their distribution differs from the inference distribution and thus exhibits sample selection bias (SSB) that affects model performance. Existing studies on SSB mainly employ sample re-weighting techniques which suffer from high variance and poor model calibration. Another line of work relies on costly uniform data that is inadequate to train industrial models. Thus mitigating SSB in industrial models with a uniform-data-free framework is worth exploring. Fortunately, many platforms display mixed results of organic items (i.e., recommendations) and sponsored items (i.e., ads) to users, where impressions of ads and recommendations are selected by different systems but share the same user decision rationales. Based on the above characteristics, we propose to leverage recommendations samples as a free lunch to mitigate SSB for ads CTR model (Rec4Ad). After elaborating data augmentation, Rec4Ad learns disentangled representations with alignment and decorrelation modules for enhancement. When deployed in Taobao display advertising system, Rec4Ad achieves substantial gains in key business metrics, with a lift of up to +6.6\% CTR and +2.9\% RPM

    Determination of 27 fragrances in cosmetics and perfume raw materials by gas chromatography-mass spectrometry

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    Treatment of Synthetic Ammonium Sulfate Wastewater by Mixed Culture of Chlorella pyrenoidosa and Enriched Nitrobacteria

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    Ammonium sulfate wastewater can cause eutrophication and black odor of water body. Although ammonia nitrogen can be used as nutrient of microalgae, high ammonia nitrogen levels could inhibit the growth of microalgae. Nitrobacteria can transform ammonia nitrogen into nitrate nitrogen. In this study, mono Chlorella pyrenoidosa culture (mono-C.py), synchronous mixed culture (mixed-a), and asynchronous mixed culture (mixed-b) systems were examined for their ability to treat ammonium sulfate wastewater. Nitrogen removal rate of mixed-b at the end of culture (52.96%) was higher than that of the mono-C.py (46.37%) and the mixed-a (39.11%). Higher total suspended solid concentration (2.40 g/L), crude protein yield (0.76 g/L), and heating value yield (35.73 kJ/L) were obtained in mixed-b, meanwhile with excellent settlement performance (91.43 +/- 0.51%). Mechanism analysis of settlement showed that the relative abundance of floc-forming-related bacteria Sphingopyxis and Acidovorax were increased generally, while nitrification/denitrifying members were decreased in mixed-b along with the culture proceeding

    The joint effect of ammonium and pH on the growth of Chlorella vulgaris and ammonium removal in artificial liquid digestate

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    Although ammonium containing digestate is an ideal alternative medium for microalgae cultivation, high ammonium or unfavorable pH may inhibit microalgal growth. In this study, the joint effect of ammonium and pH on the growth of C. vulgaris and nutrient removal in artificial digestate was investigated. Our results show that ammonium and pH both affected algal growth, but free ammonia (FA) was the main actual inhibitory factor. Algal specific growth rate presented a negative correlation with FA and their relationship was well fitted by a linear regression model. Microalgal growth was little affected below 36.8 mg L-1 FA, while the obvious inhibition occurred at 184 mg L-1 FA (EC50), indicating a high tolerance to FA. Ammonium removal was well described by a first-order kinetics model. FA stress stimulated the production of extracellular organic matters (EOMs), which was good for micmalgae adaptation but adverse to pollutant removal

    Performance of a microalgal-bacterial consortium system for the treatment of dairy-derived liquid digestate and biomass production

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    To enhance the treatment performance of dairy-derived liquid digestate (DLD) using microalgal-bacterial consortium system composed of Chlorella vulgaris and indigenous bacteria (CV), activated sludge was introduced to form a new microalgal-bacterial consortium system (Co-culture). The activated sludge shortened the lag phase and increased the specific growth rate of C. vulgaris (0.56 d(-1)). The biomass yield in the Co-culture was 2.72 g L-1, which was lower than that in the CV (3.24 g L-1), but the Co-culture had an improved COD (chemical oxygen demand) removal (25.26%) compared to the CV (13.59%). Quantitative PCR and metagenomic analyses demonstrated that microalgae also promoted bacterial growth, but influenced differently on the bacterial communities of indigenous bacteria and activated sludge. Compared with indigenous bacteria, activated sludge was more prone to forming a favorable symbiosis with C. vulgaris. These findings contribute to the construction of efficient microalgal-bacterial consortium system in wastewater treatment

    Cultivation of Chlorella vulgaris on unsterilized dairy-derived liquid digestate for simultaneous biofuels feedstock production and pollutant removal

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    In order to assess viability of microalgae cultivation using unsterilized dairy-derived liquid digestate (DLD) for simultaneous biofuels feedstock production and contaminant removal, four DLD concentrations (25%, 50%, 75% and 100%) were used to grow Chlorella vulgaris in batch photobioreactors (PBRs). The 25% DLD was an ideal alternative medium in that high growth rate (0.69 d(-1)), high lipid productivity (112.9 mg L-1 d(-1)) as well as high nutrient removal were attained. The high DLD concentration caused inhibition of microalgal growth, where COD was more inhibitive than ammonium. The presence of bacteria did not influence microalgae production because of limited growth. Microalgal growth reduced the richness and diversity of bacterial community. Furthermore, the species of Bacteroidetes, Candidatus Saccharibacteria, and Chlamydiae rather than Proteobacteria benefited microalgal-bacterial symbiosis. These findings contribute to better application of microalgal-bacterial system for large-scale microalgae cultivation as well as environmental sustainability

    Enhancement of the energy yield from microalgae via enzymatic pretreatment and anaerobic co-digestion

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    Microalgae have been considered as one of the most promising biomass resources for the generation of biodiesel, whereas algal residues (AR) after lipid extraction need to be utilized to enhance its economy and sustainability. This study proposed an integrated biochemical method for microalgae Chlorella sp. processing to achieve a high energy yield (12.33 KJ g(-1)VS). The microalgae were first pretreated with mixed enzymes for lipid extraction. After that, the AR obtained from the lipid extraction of microalgae was co-digested with energy grass (GR) Pennisetum hybrid for methane production. Results indicated that the lipid yield from microalgae was enhanced by 54.45% with mixed enzymes pretreatment of cellulase, xynalase and pectinase compared to that without enzymatic pretreatment. The maximal methane yield (207.35 +/- 15.66 mLCH(4) g(-1)VS) of AR and GR co-digestion was obtained with the AR and GR VS ratio at 1:3. Energy yields evaluation based on the biodiesel and methane yields indicated that the energy yield for this two-step combined method was 169% higher than that for biodiesel alone. Thus, the proposed integrated approach was verified to be applicable for microalgae processing for energy production. (C) 2018 Published by Elsevier Ltd

    Threshold for Relationship between Vitamin D and Parathyroid Hormone in Chinese Women of Childbearing Age

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    Background: The aim of this study was to assess the relationship between serum 25-hydroxyvitamin D [25(OH)D] and serum intact parathyroid hormone (PTH) in Chinese childbearing women, and to estimate the optimum threshold of 25(OH)D that maximally inhibits the PTH, which is considered to be the optimal status for vitamin D sufficiency. Methods: Serum samples were selected from the biological samples’ bank of the Chinese Chronic Diseases and Nutrition Survey (CCDNS) 2015. The serum 25(OH)D concentration was determined by liquid chromatography tandem mass spectrometry and the serum PTH was determined by electronic chemiluminescence. Simple linear and partial correlation analysis, locally weighted regression smooth scatterplot (LOESS), nonlinear least squares estimation (NLS), and segmented regression (SR) were utilized to estimate the relationship of 25(OH)D and PTH, and to determine the threshold of 25(OH)D. Results: A total of 1568 serum samples of 25(OH)D concentration and PTH concentration were analyzed. A significant inverse relationship between 25(OH)D and PTH concentration was observed below 15.25 (14.22–16.28) ng/mL, and PTH decreased slowly with the increase of 25(OH)D above 16.75 (15.43–18.06) ng/mL after adjusting by age, latitude, city type, season, corrected calcium, and phosphorus. A very short plateau of PTH was found at 15.25 ng/mL and 16.75 ng/mL in terms of 25(OH)D according to LOESS, NLS, and SR. Conclusions: The serum 25(OH)D was negatively correlated with the serum PTH. The threshold of VitD sufficiency was found in the range of 14.22–18.06 ng/mL in terms of serum 25(OH)D concentration for Chinese childbearing women aged 18–44 years old
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