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1.
Elmén J Lindow M Silahtaroglu A Bak M Christensen M Lind-Thomsen A Hedtjärn M Hansen JB Hansen HF Straarup EM McCullagh K Kearney P Kauppinen S 《Nucleic acids research》2008,36(4):1153-1162
MicroRNA-122 (miR-122) is an abundant liver-specific miRNA, implicated in fatty acid and cholesterol metabolism as well as hepatitis C viral replication. Here, we report that a systemically administered 16-nt, unconjugated LNA (locked nucleic acid)-antimiR oligonucleotide complementary to the 5′ end of miR-122 leads to specific, dose-dependent silencing of miR-122 and shows no hepatotoxicity in mice. Antagonism of miR-122 is due to formation of stable heteroduplexes between the LNA-antimiR and miR-122 as detected by northern analysis. Fluorescence in situ hybridization demonstrated uptake of the LNA-antimiR in mouse liver cells, which was accompanied by markedly reduced hybridization signals for mature miR-122 in treated mice. Functional antagonism of miR-122 was inferred from a low cholesterol phenotype and de-repression within 24 h of 199 liver mRNAs showing significant enrichment for miR-122 seed matches in their 3′ UTRs. Expression profiling extended to 3 weeks after the last LNA-antimiR dose revealed that most of the changes in liver gene expression were normalized to saline control levels coinciding with normalized miR-122 and plasma cholesterol levels. Combined, these data suggest that miRNA antagonists comprised of LNA are valuable tools for identifying miRNA targets in vivo and for studying the biological role of miRNAs and miRNA-associated gene-regulatory networks in a physiological context. 相似文献
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Shared patterns of species turnover between seaweeds and seed plants break down at increasing distances from the sea 下载免费PDF全文
Carlos F. D. Gurgel Thomas Wernberg Mads S. Thomsen Bayden D. Russell Paul Adam Jonathan M. Waters Sean D. Connell 《Ecology and evolution》2014,4(1):27-34
We tested for correlations in the degree of spatial similarity between algal and terrestrial plants communities along 5500 km of temperate Australian coastline and whether the strength of correlation weakens with increasing distance from the coast. We identified strong correlations between macroalgal and terrestrial plant communities within the first 100 km from shore, where the strength of these marine–terrestrial correlations indeed weakens with increasing distance inland. As such, our results suggest that marine‐driven community homogenization processes decompose with increasing distance from the shore toward inland. We speculate that the proximity to the marine environment produces lower levels of community turnover on land, and this effect decreases progressively farther inland. Our analysis suggests underlying ecological and evolutionary processes that give rise to continental‐scale biogeographic influence from sea to land. 相似文献
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Bennedsen M Stuer-Lauridsen B Danielsen M Johansen E 《Applied and environmental microbiology》2011,77(8):2785-2787
Second-generation genome sequencing and alignment of the resulting reads to in silico genomes containing antimicrobial resistance and virulence factor genes were used to screen for undesirable genes in 28 strains which could be used in human nutrition. No virulence factor genes were detected, while several isolates contained antimicrobial resistance genes. 相似文献
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Alastair B. Ross Cecilia Svelander Otto I. Savolainen Mads Vendelbo Lind John P. Kirwan Isabelle Breton Jean-Philippe Godin Ann-Sofie Sandberg 《Analytical biochemistry》2016
Plasma alkylresorcinols are increasingly analyzed in cohort studies to improve estimates of whole grain intake and their relationship with disease incidence. Current methods require large volumes of solvent (>10 ml/sample) and have relatively low daily sample throughput. We tested five different supported extraction methods for extracting alkylresorcinols from plasma and improved a normal-phase liquid chromatography coupled to a tandem mass spectrometer method to reduce sample analysis time. The method was validated and compared with gas chromatography–mass spectrometry analysis. Sample preparation with HybridSPE supported extraction was most effective for alkylresorcinol extraction, with recoveries of 77–82% from 100 μl of plasma. The use of 96-well plates allowed extraction of 160 samples per day. Using a 5-cm NH2 column and heptane reduced run times to 3 min. The new method had a limit of detection and limit of quantification equivalent to 1.1–1.8 nmol/L and 3.5–6.1 nmol/L plasma, respectively, for the different alkylresorcinol homologues. Accuracy was 93–105%, and intra- and inter-batch precision values were 4–18% across different plasma concentrations. This method makes it possible to quantify plasma alkylresorcinols in 100 μl of plasma at a rate of at least 160 samples per day without the need for large volumes of organic solvents. 相似文献
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Jayde A. Gawthorne L. Evan Reddick Snezhana N. Akpunarlieva Katherine S. H. Beckham John M. Christie Neal M. Alto Mads Gabrielsen Andrew J. Roe 《PloS one》2012,7(12)
In this work, we describe the utility of Light, Oxygen, or Voltage-sensing (LOV) flavoprotein domains from plant phototropins as a reporter for protein expression and function. Specifically, we used iLOV, an enhanced and more photostable variant of LOV. A pET-based plasmid for protein expression was constructed, encoding a C terminal iLOV-octahistidine (His8)-tag and a HRV 3C protease cleavage recognition site. Ten different proteins, with various sub-cellular locations, were cloned into the plasmid, creating iLOV-His8 tag fusions. To test protein expression and how iLOV could be used as a reporter, the proteins were expressed in three different cell lines, in four different culture media, at two different temperatures. To establish whether the presence of the iLOV tag could have an impact on the functionality, one of the proteins, EspG, was over-expressed and purified. EspG is an “effector” protein normally produced by enterohemorrhagic E. coli strains and “injected” into host cells via the T3SS. We tested functionality of EspG-iLOV fusion by performing functional studies of EspG in mammalian host cells. When EspG-iLOV was microinjected into the host cell, the Golgi apparatus was completely disrupted as had previously been observed for EspG. 相似文献
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Camilla Schmidt Morgen Benjamin Rokholm Carina Sj?berg Brixval Camilla Schou Andersen Lise Geisler Andersen Mette Rasmussen Anne-Marie Nybo Andersen Pernille Due Thorkild I. A. S?rensen 《PloS one》2013,8(7)