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100 result(s) for "Cai, Danfeng"
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Phase separation of YAP reorganizes genome topology for long-term YAP target gene expression
Yes-associated protein (YAP) is a transcriptional co-activator that regulates cell proliferation and survival by binding to a select set of enhancers for target gene activation. How YAP coordinates these transcriptional responses is unknown. Here, we demonstrate that YAP forms liquid-like condensates in the nucleus. Formed within seconds of hyperosmotic stress, YAP condensates compartmentalized the YAP transcription factor TEAD1 and other YAP-related co-activators, including TAZ, and subsequently induced the transcription of YAP-specific proliferation genes. Super-resolution imaging using assay for transposase-accessible chromatin with photoactivated localization microscopy revealed that the YAP nuclear condensates were areas enriched in accessible chromatin domains organized as super-enhancers. Initially devoid of RNA polymerase II, the accessible chromatin domains later acquired RNA polymerase II, transcribing RNA. The removal of the intrinsically-disordered YAP transcription activation domain prevented the formation of YAP condensates and diminished downstream YAP signalling. Thus, dynamic changes in genome organization and gene activation during YAP reprogramming is mediated by liquid–liquid phase separation. Cai et al. show that YAP forms liquid-like condensates in the nucleus that compartmentalize YAP’s DNA binding cofactors and transcription co-activators to induce transcription of YAP-specific proliferation genes.
Modeling and analysis of collective cell migration in an in vivo three-dimensional environment
A long-standing question in collective cell migration has been what might be the relative advantage of forming a cluster over migrating individually. Does an increase in the size of a collectively migrating group of cells enable them to sample the chemical gradient over a greater distance because the difference between front and rear of a cluster would be greater than for single cells? We combined theoretical modeling with experiments to study collective migration of the border cells in-between nurse cells in the Drosophila egg chamber. We discovered that cluster size is positively correlated with migration speed, up to a particular point above which speed plummets. This may be due to the effect of viscous drag from surrounding nurse cells together with confinement of all of the cells within a stiff extracellular matrix. The model predicts no relationship between cluster size and velocity for cells moving on a flat surface, in contrast to movement within a 3D environment. Our analyses also suggest that the overall chemoattractant profile in the egg chamber is likely to be exponential, with the highest concentration in the oocyte. These findings provide insights into collective chemotaxis by combining theoretical modeling with experimentation.
The joy of scientific discoveries and interactions
Danfeng Cai, an assistant professor in the Department of Biochemistry and Molecular Biology at the Johns Hopkins Bloomberg School of Public Health, discusses her career path, including her work on the biomolecular condensation of YAP, and her excitement in her ongoing work on transcriptional condensates.
Indirect Competitive Enzyme-Linked Immunosorbent Assay Based on Broad-Spectrum Antibody for Simultaneous Determination of Thirteen Fluoroquinolone Antibiotics in Rana catesbeianus
Fluoroquinolone (FQ) is a type of widely used antibiotic in agriculture and aquaculture, and exposure to low doses of FQs may result in the transfer of resistance between animal and human pathogens. Based on the optimization of the operating parameters, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) standard curve was constructed for the simultaneous detection of 13 FQs, including enrofloxacin (ENR), ciprofloxacin (CIP), sarafloxacin (SAR), ofloxacin (OFL), norfloxacin (NOR), pefloxacin mesylate (PM), pefloxacin (PEF), enoxacin (ENX), marbofloxacin (MAR), fleroxacin (FLE), lomefloxacin (LOM), danofloxacin (DAN), and difloxacin (DIF). The limit of detection (LOD, computed as IC10) and sensitivity (IC50) of the ic-ELISA for ENR were 0.59 μg/L and 19.23 μg/L, respectively. The precision and dependability of the detection results of this ic-ELISA were properly verified by HPLC in Rana catesbeianus samples. This indicated that the established ic-ELISA approach could be utilized to determine the FQs in Rana catesbeianus. In addition, this ic-ELISA, based on a broad-spectrum antibody, provides a technical reference and potential strategy for an immunoassay of hazard factors with similar structure.
METTL3 facilitates the translation of CircSIK2 during chicken myogenesis in an m6A dependent manner
Circular RNAs (circRNAs) are generally considered a new class of non-coding RNA (ncRNA) that frequently appears in the eukaryotic transcriptome. In principle, circRNAs may encode proteins, as some of them are generated from exons and possess elements for internal ribosome entry. Circular RNAs have the potential to serve as an unexplored reservoir for the generation of novel proteins, yet the identification of coding-circRNAs is a daunting task. In this study, we developed a specialized strategy for the discovery of coding-circRNA by combining RNA sequencing, ribosome profiling, and mass spectrometry to find a multitude of circRNAs translated in vivo . A total of 40,084 circRNAs were found in chicken myoblasts and myotubes, and 15,332 circRNAs had a predicted open reading frame (ORF). Via ribosome footprints, we discovered that a group of circRNAs (4,069) was associated with translating ribosomes (ribo-circRNAs). Moreover, a total of 3,927 circRNAs with an infinite ORF were discovered, and 860 of them were associated with translating ribosome (ribo-no-stop-codon circRNAs). Mass spectrometry found 5 specific peptides spectra spanning a back-splice junction of circRNAs. circSIK2, one of the ribo-circRNAs, could be methylated by METTL3 and translated into SIK2-176aa, thus promoting the proliferation and differentiation of myoblasts and muscle hypertrophy. Our results suggest that many circRNAs were translating during chicken myogenesis, and METTL3 could enhance the translation of circSIK2. To the best of our knowledge, only two circRNAs translation events have been reported to be mediated by m 6 A. Our research would represent the third such event, and the first documented instance of a translatable circRNA in poultry.
DNA methylation in poultry: a review
As an important epigenetic modification, DNA methylation is involved in many biological processes such as animal cell differentiation, embryonic development, genomic imprinting and sex chromosome inactivation. As DNA methylation sequencing becomes more sophisticated, it becomes possible to use it to solve more zoological problems. This paper reviews the characteristics of DNA methylation, with emphasis on the research and application of DNA methylation in poultry.
Determination Methods of the Risk Factors in Food Based on Nanozymes: A Review
Food safety issues caused by foodborne pathogens, chemical pollutants, and heavy metals have aroused widespread concern because they are closely related to human health. Nanozyme-based biosensors have excellent characteristics such as high sensitivity, selectivity, and cost-effectiveness and have been used to detect the risk factors in foods. In this work, the common detection methods for pathogenic microorganisms, toxins, heavy metals, pesticide residues, veterinary drugs, and illegal additives are firstly reviewed. Then, the principles and applications of immunosensors based on various nanozymes are reviewed and explained. Applying nanozymes to the detection of pathogenic bacteria holds great potential for real-time evaluation and detection protocols for food risk factors.
Does Participation in Aquaculture Cooperatives Increase Farmers′ Profit and Output?
Individual aquaculture farmers in developing countries play an important role in ensuring food security. This study uses survey data from aquaculture households in Rongcheng and Xiangshan cities in China to explore the impact of cooperative participation on the benefits to the aquaculture households. The empirical results show that the participation of aquaculture farmers in cooperatives has effectively increased their net profit and output per unit area. On average, participating in cooperatives increased the net profit and output per unit area of farmers by approximately 15.55% and 11.47%, respectively. The test results of the mechanism show that the information services, technical training, and product sales guidance provided by the cooperatives have increased the net profit of the farmers. At the same time, the information services and product sales guidance provided by cooperatives are important reasons for the increase in the output per unit area.
A Smartphone Colorimetric Sensor Based on Pt@Au Nanozyme for Visual and Quantitative Detection of Omethoate
A smartphone colorimetric sensor based on the Pt@Au nanozyme was successfully developed for the visual and quantitative detection of omethoate in fruit and vegetables. The anti-omethoate antibody was conjugated on the surface of the Pt@Au nanozyme as a catalytic functional signal probe, and coating antigen conjugated on the surface of magnetic polystyrene microspheres (MPMs) was used as a separation capture probe. In the sensing system, when the catalytic functional signal probe was combined with a separation capture probe containing no omethoate, the visible blue color appeared with the addition of tetramethylbenzidine (TMB) chromogenic solution, and the maximum B value of the sensing system was obtained via the smartphone. With increasing concentrations of omethoate, the visualization of the sensing system decreased, and the B-value obtained via the smartphone dropped. Under optimal detection conditions, the omethoate could be detected in a linear range of 0.5–50 μg/L (R2 = 0.9965), with a detection limit of 0.01 μg/L. The accuracy and reliability of the detection results of this colorimetric sensor were successfully confirmed by enzyme linked immunosorbent assay (ELISA) and gas chromatography. This colorimetric sensor provides a technical reference and potential strategy for the immunoassay of hazard factors in resource-scarce laboratories.
CircMGA Depresses Myoblast Proliferation and Promotes Myotube Formation through miR-144-5p/FAP Signal
Circular RNAs are endogenous and abundant in skeletal muscle, and may not only be involved in regulating gene expression in a variety of ways, but also function as important regulators in poultry muscle development. Our previous research found that circMGA was differentially expressed during chicken muscle embryo development; however, as a novel circular RNA, the regulating mechanism of circMGA in myogenesis has never been studied before. In this study, we aimed to investigate the functional roles and related molecular mechanisms of circMGA in chicken primary myoblast cells. CircMGA originated from the exon 13–14 of MGA gene, was differentially expressed during embryo development and myogenesis differentiation, and could inhibit myoblast cell proliferation by repressing cell cycle related genes and promote myotube formation through MyoD and MyHC. Biotin-labeled miRNA pulldown assay and luciferase reporter assay result showed that miR-144-5p could directly target circMGA and FAP, indicating that there could be a competing endogenous RNA mechanism between circMGA and FAP. In function, miR-144-5p showed opposite regulation in myoblast cell with circMGA and FAP, just as expected. circMGA co-transfected with miR-144-5p or si-FAP could effectively eliminate the inhibition of miR-144-5p on myoblast proliferation and differentiation. In conclusion, we found a novel circRNA, named circMGA, which generated from the 13–14 exon of the MGA gene, and could inhibit myoblast proliferation and promote myotube formation by acting as the sponge of miR-144-5p and through miR-144-5p/FAP signal. Moreover, circMGA could effectively eliminate the inhibition of miR-144-5p on myoblast differentiation, thus releasing FAP and promoting myotube formation.