743 research outputs found

    Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins

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    We propose the concept of universal fiducials based on a set of pre-made semi-synthetic antibodies (sABs) generated by customized phage display selections against the fusion protein BRIL, an engineered variant of apocytochrome b562a. These sABs can bind to BRIL fused either into the loops or termini of different GPCRs, ion channels, receptors and transporters without disrupting their structure. A crystal structure of BRIL in complex with an affinity-matured sAB (BAG2) that bound to all systems tested delineates the footprint of interaction. Negative stain and cryoEM data of several examples of BRIL-membrane protein chimera highlight the effectiveness of the sABs as universal fiducial marks. Taken together with a cryoEM structure of sAB bound human nicotinic acetylcholine receptor, this work demonstrates that these anti-BRIL sABs can greatly enhance the particle properties leading to improved cryoEM outcomes, especially for challenging membrane proteins

    Accumulation, temporal variation, source apportionment and risk assessment of heavy metals in agricultural soils from the middle reaches of Fenhe River basin, North China

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    The Fenhe River basin is the main agricultural and industrial developed area in Shanxi province, China. In recent years, agricultural non-point source pollution in the Fenhe River basin intensified, threatening soil quality and safety in the area. Accumulation of eight heavy metals (HMs) including chromium (Cr), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) has been detected in soil samples from 50 agricultural sites (0-20 cm) from the middle reaches of the Fenhe River basin. The ecological and human health risk and potential sources of the eight HMs were investigated. In addition, the human health and ecological risks imposed by the possible sources of the eight HMs were quantitatively apportioned. The enrichment factor (EF) values of Cr, Ni, Cu, Pb and Zn were lower than 2, indicating minimal enrichment, while values for As, Cd and Hg were between 2 and 5, exhibiting moderate enrichment. Temporal variation analysis suggested that most HMs in the study area exhibited low concentrations after 2015, except As. The potential ecological risk index was 174.09, indicating low ecological risk. The total hazard index and cancer risk values were 0.395 and 5.35 x 10(-4) for adults and 2.75 and 3.63 x 10(-4) for children, indicating the accepted standard levels were exceeded for non-carcinogenic risk for children and carcinogenic risks for both adults and children. Four potential sources were identified: (1) natural sources, (2) farming activities, (3) coal combustion, and (4) exhaust emissions. Natural sources represented the largest contributor to ecological risk, accounting for 57.42% of the total. Coal combustion was the major contributor to human health risks, accounting for 43.27% and 43.73% of the total non-carcinogenic risk and carcinogenic risk for adults, respectively, and 42.72% and 43.88% for children, respectively

    Enabling multi-level relevance feedback on PubMed by integrating rank learning into DBMS

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    Background: Finding relevant articles from PubMed is challenging because it is hard to express the user's specific intention in the given query interface, and a keyword query typically retrieves a large number of results. Researchers have applied machine learning techniques to find relevant articles by ranking the articles according to the learned relevance function. However, the process of learning and ranking is usually done offline without integrated with the keyword queries, and the users have to provide a large amount of training documents to get a reasonable learning accuracy. This paper proposes a novel multi-level relevance feedback system for PubMed, called RefMed, which supports both ad-hoc keyword queries and a multi-level relevance feedback in real time on PubMed. Results: RefMed supports a multi-level relevance feedback by using the RankSVM as the learning method, and thus it achieves higher accuracy with less feedback. RefMed "tightly" integrates the RankSVM into RDBMS to support both keyword queries and the multi-level relevance feedback in real time; the tight coupling of the RankSVM and DBMS substantially improves the processing time. An efficient parameter selection method for the RankSVM is also proposed, which tunes the RankSVM parameter without performing validation. Thereby, RefMed achieves a high learning accuracy in real time without performing a validation process. RefMed is accessible at http://dm.postech.ac.kr/refmed. Conclusions: RefMed is the first multi-level relevance feedback system for PubMed, which achieves a high accuracy with less feedback. It effectively learns an accurate relevance function from the user's feedback and efficiently processes the function to return relevant articles in real time.1114Nsciescopu

    The critical role of the operating conditions on the Fenton oxidation of 2-chlorophenol: assessment of PCDD/Fs formation

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    This work assesses the influence of the operating conditions H2O2dose (20 or 100% of the stoichiometric amount), temperature (20 or 70◩C), and the presence of chloride in the oxidation medium in the formation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) during Fenton treatment of aqueous samples of 2-chlorophenol, 2-CP, one of the strongest precursor of PCDD/Fs. After 4 h of oxidation in the experiments carried out with 20% H2O2chlorinated phenoxyphenols and biphenyls, which are intermediates in PCDD/Fs formation, as well as PCDD/Fs were observed, resulting in concentrations11 times higher than in the untreated sample. Additionally, when NaCl was also present in the reaction medium, PCDD/Fs were formed at higher extent, with a total concentration 74.4 times higher than in the untreated 2-CP solution. Results depicted a preferential formation of PCDFs over PCDDs, with dominance of lower chlorinated PCDD/Fs (tetra and penta-PCDD/Fs). Besides, the formation of the most toxic PCDD/Fs congeners (2,3,7,8-PCDD/Fs) was not favored under the operating conditions used in this work.Financial support from the projects CTQ2011-25262, CTQ2008-05545 and CTQ2008-00690 (Ministerio de Economía y Competitividad – MINECO (SPAIN) and Fondo Europeo de Desarrollo Regional – FEDER) is gratefully acknowledged

    The LDL Receptor Clustering Motif Interacts with the Clathrin Terminal Domain in a Reverse Turn Conformation

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    Previously the hexapeptide motif FXNPXY807 in the cytoplasmic tail of the LDL receptor was shown to be essential for clustering in clathrin-coated pits. We used nuclear magnetic resonance line-broadening and transferred nuclear Overhauser effect measurements to identify the molecule in the clathrin lattice that interacts with this hexapeptide, and determined the structure of the bound motif. The wild-type peptide bound in a single conformation with a reverse turn at residues NPVY. Tyr807Ser, a peptide that harbors a mutation that disrupts receptor clustering, displayed markedly reduced interactions. Clustering motif peptides interacted with clathrin cages assembled in the presence or absence of AP2, with recombinant clathrin terminal domains, but not with clathrin hubs. The identification of terminal domains as the primary site of interaction for FXNPXY807 suggests that adaptor molecules are not required for receptor-mediated endocytosis of LDL, and that at least two different tyrosine-based internalization motifs exist for clustering receptors in coated pits

    A global model of meteoric sodium

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    A global model of sodium in the mesosphere and lower thermosphere has been developed within the framework of the National Center for Atmospheric Research's Whole Atmosphere Community Climate Model (WACCM). The standard fully interactive WACCM chemistry module has been augmented with a chemistry scheme that includes nine neutral and ionized sodium species. Meteoric ablation provides the source of sodium in the model and is represented as a combination of a meteoroid input function (MIF) and a parameterized ablation model. The MIF provides the seasonally and latitudinally varying meteoric flux which is modeled taking into consideration the astronomical origins of sporadic meteors and considers variations in particle entry angle, velocity, mass, and the differential ablation of the chemical constituents. WACCM simulations show large variations in the sodium constituents over time scales from days to months. Seasonality of sodium constituents is strongly affected by variations in the MIF and transport via the mean meridional wind. In particular, the summer to winter hemisphere flow leads to the highest sodium species concentrations and loss rates occurring over the winter pole. In the Northern Hemisphere, this winter maximum can be dramatically affected by stratospheric sudden warmings. Simulations of the January 2009 major warming event show that it caused a short-term decrease in the sodium column over the polar cap that was followed by a factor of 3 increase in the following weeks. Overall, the modeled distribution of atomic sodium in WACCM agrees well with both ground-based and satellite observations. Given the strong sensitivity of the sodium layer to dynamical motions, reproducing its variability provides a stringent test of global models and should help to constrain key atmospheric variables in this poorly sampled region of the atmosphere. Key Points The first global model of mesospheric sodium has been developed Model includes new description of the meteoroid input function Meridional winds and stratospheric sudden warnings affect the sodium column

    Phytochemical content and antioxidant activity of grapefruit (Star Ruby): A comparison between fresh freeze-dried fruits and different powder formulations

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    [EN] Different grapefruit powders obtained by freeze drying and spray drying with prior addition of shell materials (arabic gum and bamboo fiber) were studied in order to evaluate the effect of these preservation processes on the retention of antioxidants, in comparison with the freeze-dried fruit with no carriers added. Freeze-dried samples showed above 90% retention of these phytochemicals, while spray-dried samples presented good retention of vitamins but a sharp decrease in of phenolic compounds. Pearson's correlation analysis showed that the most significant contribution to DPPH scavenging activity and inhibition of β-carotene bleaching was provided by phenolic compounds, mostly flavonoids, while the contribution to the reducing power was due to ascorbic acid and α-tocopherol. Therefore, the loss of these compounds in the spray-dried samples resulted in products with lower antioxidant activity. Naringin and narirutin were the major phenolic compounds in all grapefruit samples, although other flavanones present in lower concentration, like hesperidin, neohesperidin didymin, poncirin or melitidin, also showed high correlations with the antioxidant value of the samples.The authors thank the Ministerio de Economia y Competitividad for the financial support given throughout the Project AGL 2012-39103, and to the Foundation for Science and Technology and FEDER under Program PT2020 for financial support to CIMO (UID/AGR/00690/2013), and L. Barros (SFRH/BPD/107855/2015) grant. To POCI-01-0145-FEDER-006984 (LA LSRE-LCM), funded by ERDF, through POCI-COMPETE2020 and FCT.Agudelo-Sterling, C.; Barros, L.; Santos-Buelga, C.; MartĂ­nez Navarrete, N.; Ferreira, ICFR. (2017). Phytochemical content and antioxidant activity of grapefruit (Star Ruby): A comparison between fresh freeze-dried fruits and different powder formulations. LWT - Food Science and Technology. 80:106-112. https://doi.org/10.1016/j.lwt.2017.02.006S1061128
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