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21.
Felipe Romero-Saavedra Diana Laverde Dominique Wobser Charlotte Michaux Aurélie Budin-Verneuil Benoit Bernay Abdellah Benachour Axel Hartke Johannes Huebner 《PloS one》2014,9(11)
Infections by opportunistic bacteria have significant contributions to morbidity and mortality of hospitalized patients and also lead to high expenses in healthcare. In this setting, one of the major clinical problems is caused by Gram-positive bacteria such as enterococci and staphylococci. In this study we extract, purify, identify and characterize immunogenic surface-exposed proteins present in the vancomycin resistant enterococci (VRE) strain Enterococcus faecium E155 using three different extraction methods: trypsin shaving, biotinylation and elution at high pH. Proteomic profiling was carried out by gel-free and gel-nanoLC-MS/MS analyses. The total proteins found with each method were 390 by the trypsin shaving, 329 by the elution at high pH, and 45 using biotinylation. An exclusively extracytoplasmic localization was predicted in 39 (10%) by trypsin shaving, in 47 (15%) by elution at high pH, and 27 (63%) by biotinylation. Comparison between the three extraction methods by Venn diagram and subcellular localization predictors (CELLO v.2.5 and Gpos-mPLoc) allowed us to identify six proteins that are most likely surface-exposed: the SCP-like extracellular protein, a low affinity penicillin-binding protein 5 (PBP5), a basic membrane lipoprotein, a peptidoglycan-binding protein LysM (LysM), a D-alanyl-D-alanine carboxypeptidase (DdcP) and the peptidyl-prolyl cis-trans isomerase (PpiC). Due to their close relationship with the peptidoglycan, we chose PBP5, LysM, DdcP and PpiC to test their potential as vaccine candidates. These putative surface-exposed proteins were overexpressed in Escherichia coli and purified. Rabbit polyclonal antibodies raised against the purified proteins were able to induce specific opsonic antibodies that mediated killing of the homologous strain E. faecium E155 as well as clinical strains E. faecium E1162, Enterococcus faecalis 12030, type 2 and type 5. Passive immunization with rabbit antibodies raised against these proteins reduced significantly the colony counts of E. faecium E155 in mice, indicating the effectiveness of these surface-related proteins as promising vaccine candidates to target different enterococcal pathogens. 相似文献
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Abdellah Barakate James D. Higgins Sebastian Vivera Jennifer Stephens Ruth M. Perry Luke Ramsay Isabelle Colas Helena Oakey Robbie Waugh F. Chris H. Franklin Susan J. Armstrong Claire Halpin 《The Plant cell》2014,26(2):729-740
In many cereal crops, meiotic crossovers predominantly occur toward the ends of
chromosomes and 30 to 50% of genes rarely recombine. This limits the exploitation of
genetic variation by plant breeding. Previous reports demonstrate that chiasma
frequency can be manipulated in plants by depletion of the synaptonemal complex
protein ZIPPER1 (ZYP1) but conflict as to the direction of change, with fewer
chiasmata reported in Arabidopsis thaliana and more crossovers
reported for rice (Oryza sativa). Here, we use RNA interference
(RNAi) to reduce the amount of ZYP1 in
barley (Hordeum vulgare) to only 2 to 17% of normal zygotene levels.
In the ZYP1RNAi lines, fewer than half of the chromosome
pairs formed bivalents at metaphase and many univalents were observed, leading to
chromosome nondisjunction and semisterility. The number of chiasmata per cell was
reduced from 14 in control plants to three to four in the ZYP1-depleted lines,
although the localization of residual chiasmata was not affected. DNA double-strand
break formation appeared normal, but the recombination pathway was defective at later
stages. A meiotic time course revealed a 12-h delay in prophase I progression to the
first labeled tetrads. Barley ZYP1 appears to function similarly to ZIP1/ZYP1 in
yeast and Arabidopsis, with an opposite effect on crossover number
to ZEP1 in rice, another member of the Poaceae. 相似文献
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One hundred bacteria, isolated from rhizospheric soil and rhizoplane of healthy soybean plants, were assayed for antifungal activity against six Phytophthora sojae isolates. Nine of the tested bacteria inhibited the hyphal growth of P. sojae in vitro. They were subsequently evaluated for their in vitro traits and identified using the 16S rRNA gene sequences. Four of them (Paenibacillus sp.,—S1; Streptomyces sp.,—S9, S10 and S11) were further selected on the basis of their strongest antagonistic activity in vitro against P. sojae race 4, the predominant race in most growing soybean areas in Canada, and tested for their beneficial effects on soybean plants in the greenhouse. Results showed that application of bacterial strain S11 as seed coating reduced the disease severity by 57.1% and increased the root and shoot weight by and 140 and 108% respectively, in comparison to the diseased control. Overall, a positive correlation was recorded between the in vitro and in planta effects of the selected bacteria. This is promising for further application as select environmentally safe biological control agents in the protection of soybean against root rot diseases. 相似文献
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Giovanni Appendino Abdellah Ech-Chahad Alberto Minassi Luciano De Petrocellis Vincenzo Di Marzo 《Bioorganic & medicinal chemistry letters》2010,20(1):97-99
The side chain benzylic methylene is a critical element for the vanilloid activity of resiniferatoxin (2a, RTX), and introduction of branching, oxygen functions, or isosteric substitution at this center proved detrimental, with a decrease of potency of 2–3 orders of magnitude compared to the natural product. Conversely, only a modest erosion of activity was observed upon α-methylation and α-methylenation of the side chain. Surprisingly, introduction of an iodine atom in the guaiacyl moiety of the oxygen isoster 2h led to an unexpected and remarkable (>1000-fold) increase of potency, affording 2i, a compound that outperforms RTX in terms of vanilloid agonism and represents the first one-digit picomolar ligand of a TRP channel discovered to date. 相似文献
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Pavel Mader Rodrigo Mendoza-Sanchez Aman Iqbal Aiping Dong Elena Dobrovetsky Victoria B. Corless Sean K. Liew Scott R. Houliston Renato Ferreira De Freitas David Smil Carlo C. Dela Sena Steven Kennedy Diego B. Diaz Hong Wu Ludmila Dombrovski Abdellah Allali-Hassani Jinrong Min Matthieu Schapira Cheryl H. Arrowsmith 《Bioorganic & medicinal chemistry》2019,27(17):3866-3878
SET domain bifurcated protein 1 (SETDB1) is a human histone-lysine methyltransferase which is amplified in human cancers and was shown to be crucial in the growth of non-small and small cell lung carcinoma. In addition to its catalytic domain, SETDB1 harbors a unique tandem tudor domain which recognizes histone sequences containing both methylated and acetylated lysines, and likely contributes to its localization on chromatin. Using X-ray crystallography and NMR spectroscopy fragment screening approaches, we have identified the first small molecule fragment hits that bind to histone peptide binding groove of the Tandem Tudor Domain (TTD) of SETDB1. Herein, we describe the binding modes of these fragments and analogues and the biophysical characterization of key compounds. These confirmed small molecule fragments will inform the development of potent antagonists of SETDB1 interaction with histones. 相似文献
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High‐level primary clarithromycin resistance of Helicobacter pylori in Morocco: a prospective multicenter molecular study 下载免费PDF全文
27.
James D. Higgins Ruth M. Perry Abdellah Barakate Luke Ramsay Robbie Waugh Claire Halpin Susan J. Armstrong F. Chris H. Franklin 《The Plant cell》2012,24(10):4096-4109
Meiosis involves reciprocal exchange of genetic information between homologous chromosomes to generate new allelic combinations. In cereals, the distribution of genetic crossovers, cytologically visible as chiasmata, is skewed toward the distal regions of the chromosomes. However, many genes are known to lie within interstitial/proximal regions of low recombination, creating a limitation for breeders. We investigated the factors underlying the pattern of chiasma formation in barley (Hordeum vulgare) and show that chiasma distribution reflects polarization in the spatiotemporal initiation of recombination, chromosome pairing, and synapsis. Consequently, meiotic progression in distal chromosomal regions occurs in coordination with the chromatin cycles that are a conserved feature of the meiotic program. Recombination initiation in interstitial and proximal regions occurs later than distal events, is not coordinated with the cycles, and rarely progresses to form chiasmata. Early recombination initiation is spatially associated with early replicating, euchromatic DNA, which is predominately found in distal regions. We demonstrate that a modest temperature shift is sufficient to alter meiotic progression in relation to the chromosome cycles. The polarization of the meiotic processes is reduced and is accompanied by a shift in chiasma distribution with an increase in interstitial and proximal chiasmata, suggesting a potential route to modify recombination in cereals. 相似文献
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