101 research outputs found

    Analysis of Annual Available Water Resources of a Representative Basin in Upper Loess Plateau

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    Source: ICHE Conference Archive - https://mdi-de.baw.de/icheArchiv

    Optimized sample preparation for two-dimensional gel electrophoresis of soluble proteins from chicken bursa of Fabricius

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    <p>Abstract</p> <p>Background</p> <p>Two-dimensional gel electrophoresis (2-DE) is a powerful method to study protein expression and function in living organisms and diseases. This technique, however, has not been applied to avian bursa of Fabricius (BF), a central immune organ. Here, optimized 2-DE sample preparation methodologies were constructed for the chicken BF tissue. Using the optimized protocol, we performed further 2-DE analysis on a soluble protein extract from the BF of chickens infected with virulent avibirnavirus. To demonstrate the quality of the extracted proteins, several differentially expressed protein spots selected were cut from 2-DE gels and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).</p> <p>Results</p> <p>An extraction buffer containing 7 M urea, 2 M thiourea, 2% (w/v) 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), 50 mM dithiothreitol (DTT), 0.2% Bio-Lyte 3/10, 1 mM phenylmethylsulfonyl fluoride (PMSF), 20 U/ml Deoxyribonuclease I (DNase I), and 0.25 mg/ml Ribonuclease A (RNase A), combined with sonication and vortex, yielded the best 2-DE data. Relative to non-frozen immobilized pH gradient (IPG) strips, frozen IPG strips did not result in significant changes in the 2-DE patterns after isoelectric focusing (IEF). When the optimized protocol was used to analyze the spleen and thymus, as well as avibirnavirus-infected bursa, high quality 2-DE protein expression profiles were obtained. 2-DE maps of BF of chickens infected with virulent avibirnavirus were visibly different and many differentially expressed proteins were found.</p> <p>Conclusion</p> <p>These results showed that method C, in concert extraction buffer IV, was the most favorable for preparing samples for IEF and subsequent protein separation and yielded the best quality 2-DE patterns. The optimized protocol is a useful sample preparation method for comparative proteomics analysis of chicken BF tissues.</p

    Identification of hub genes in airway epithelial cells of asthma patients by WGCNA and PPI network

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    258-267Bronchial asthma is a common chronic disease of airway inflammation, high mucus secretion and airway hyper responsiveness. The pathogenetic mechanisms of asthma remain unclear. In this study, we aimed at identifying genes playing an import role in disease-related pathways in airway epithelial cells of asthma patients. Microarray data GSE41861 of asthma airway epithelial cells was used to screen differentially expressed genes (DEGs) through GEO2R analysis. The weighted gene co-expression network analysis (WGCNA) was performed to identify gene co-expression network modules in bronchial asthma. The DAVID database was then used to perform functional and pathway enrichment analysis of these DEGs. In addition, we have conducted protein-protein interaction (PPI) network of DEGs by STRING, and eventually found key genes and significant modules. A total of 315 DEGs (111 up-regulated and 204 down-regulated) were identified between severe asthma and healthy individual, which were mainly involved in pathways of cilium assembly, cilium morphogenesis, axon guidance, positive regulation of fat cell differentiation, and positive regulation of cell substrate adhesion. A total of 60 genes in the black module and green module were considered to be correlated with the severity of asthma. Combining PPI network, several key genes were identified, such as BP2RY14, PTGS1, SLC18A2, SIGLEC6, RGS13, CPA3, and HPGDS. Our findings revealed several genes that may be involved in the process of development of bronchial asthma and potentially be candidate targets for diagnosis or therapy of bronchial asthma

    Ventral Visual Pathway-Cerebellar Circuit Deficits in Alcohol Dependence: Long- and Short-Range Functional Connectivity Density Study

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    Objective: To identify the underlying intrinsic functional connectome changes in patients with alcohol dependence.Methods: A functional connectivity density (FCD) analysis was used to report on the functional connectivity changes in 24 male patients with alcohol dependence (age, 47.83 ± 6.93 years) and 24 healthy male subjects (age, 47.67 ± 6.99 years). We defined the voxels with a correlated threshold of r &gt; 0.25 inside their neighborhood (radius sphere ≤ 6 mm) as shortFCD, and radius sphere &gt; 6 mm as longFCD. We repeated the network analysis using a range of correlation r thresholds (r = 0.30, 0.35, 0.40, 0.45, 0.50, 0.6, and 0.75) to determine whether between-group differences were substantially affected by the selection of the different R-value thresholds used. A ROC curve was used to test the ability of the FCD in discriminating between the two groups. Pearson's correlation was used to evaluate the relationships between the FCD differences in brain areas and demographic characteristics.Results: The covered differences in brain areas in binarized shortFCD were larger than binarized longFCD in both groups. The intra-group FCD differences did not depend on the selection of different thresholds used. Patients with alcohol dependence were associated with the longFCD deficit in the cerebellum posterior lobe, and shortFCD deficit in the ventral system of the visual pathway and increased shortFCD in the left precentral gyrus, right salience network and right cingulate gyrus. A ROC curve demonstrated that these specific brain areas alone discriminated between the two groups with a high degree of sensitivity and specificity. In the alcohol dependence group, the cerebellum posterior lobe, visual association cortex and the salience network displayed significant correlations with demographic characteristics.Conclusions: The shortFCD analysis was more sensitive than the longFCD analysis in finding differences in the brain areas. The ventral visual pathway-cerebellar circuit deficit appeared to be altered in patients with alcohol dependence

    Dissociation of ssDNA - Single-Walled Carbon Nanotube Hybrids by Watson-Crick Base Pairing

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    The unwrapping event of ssDNA from the SWNT during the Watson-Crick base paring is investigated through electrical and optical methods, and binding energy calculations. While the ssDNA-metallic SWNT hybrid shows the p-type semiconducting property, the hybridization product recovered metallic properties. The gel electrophoresis directly verifies the result of wrapping and unwrapping events which was also reflected to the Raman shifts. Our molecular dynamics simulations and binding energy calculations provide atomistic description for the pathway to this phenomenon. This nano-physical phenomenon will open up a new approach for nano-bio sensing of specific sequences with the advantages of efficient particle-based recognition, no labeling, and direct electrical detection which can be easily realized into a microfluidic chip format.Comment: 4 pages, 4 figure
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