56 research outputs found
Графический дизайн как визуальный язык межкультурного взаимодействия
This article describes how visual graphics language as a sign system can be in contact with the audience, overcoming the language barrier. In terms of graphic design it can be available to transfer information, and even affect the viewer, causing artistic and emotional reflection.Эта статья о том, как визуальный язык графики в виде знаковой символики может входить в контакт со зрителем, преодолевая языковый барьер. На языке графического дизайна можно доступно передать информацию и даже воздействовать на зрителя, вызывая при этом художественно-эмоциональные образы
Relationships between SAV576 and SAV577.
<p>(<b>A</b>) Comparison of DNA-binding affinity of SAV576 and SAV577 with probes 1 and 577p. Each lane contained 0.15 nM labeled probe. White arrows: free probes. Black arrows: DNA-protein complexes. (<b>B</b>) Competitive EMSAs of probe 1 with His<sub>6</sub>-SAV576 and His<sub>6</sub>-SAV577 proteins. End-labeled probe 1 was incubated with the indicated concentrations of His<sub>6</sub>-SAV576 or His<sub>6</sub>-SAV577. Lanes 1 and 1′: control reactions (no protein added). Diagonal arrows: two complexes in lanes 3 and 3′. (<b>C</b>) GST pull-down assays of SAV576 and SAV577 from <i>E. coli</i> whole cell lysate. GST- and His<sub>6</sub>-tagged proteins were co-expressed in <i>E. coli</i>, lysed by sonication, and subjected to GST pull-down assay and Western blotting using anti-GST and anti-His antibodies, respectively. Lanes 1, cell lysate before IPTG induction. Lanes 2, cell lysate after induction. Lanes 3, GST pull-down.</p
Two Adjacent and Similar TetR Family Transcriptional Regulator Genes, <i>SAV577</i> and <i>SAV576</i>, Co-Regulate Avermectin Production in <i>Streptomyces avermitilis</i>
<div><p><i>Streptomyces avermitilis</i> is an important bacterial species used for industrial production of avermectins, a family of broad-spectrum anthelmintic agents. We previously identified the protein SAV576, a TetR family transcriptional regulator (TFR), as a downregulator of avermectin biosynthesis that acts by controlling transcription of its major target gene <i>SAV575</i> (which encodes cytochrome P450/NADPH-ferrihemoprotein reductase) and <i>ave</i> genes. <i>SAV577</i>, another TFR gene, encodes a SAV577 protein that displays high amino acid homology with SAV576. In this study, we examined the effect of SAV577 on avermectin production and the relationships between SAV576 and SAV577. SAV577 downregulated avermectin biosynthesis indirectly, similarly to SAV576. SAV576 and SAV577 both directly repressed <i>SAV575</i> transcription, and reciprocally repressed each other's expression. <i>SAV575</i> transcription levels in various <i>S. avermitilis</i> strains were correlated with avermectin production levels. DNase I footprinting and electrophoretic mobility shift assays indicated that SAV576 and SAV577 compete for the same binding regions, and that DNA-binding affinity of SAV576 is much stronger than that of SAV577. GST pull-down assays revealed no direct interaction between the two proteins. Taken together, these findings suggest that SAV577 regulates avermectin production in <i>S. avermitilis</i> by a mechanism similar to that of SAV576, and that the role of SAV576 is dominant over that of SAV577. This is the first report of two adjacent and similar TFR genes that co-regulate antibiotic production in <i>Streptomyces.</i></p></div
Identification of <i>SAV577</i> as a target gene of SAV576.
<p>(<b>A</b>) Real-time RT-PCR analysis of <i>SAV577</i> transcription levels for ATCC31267 (WT) and <i>SAV576</i> deletion mutant D576 grown in FM-II on days 2 and 6. Relative values were obtained using <i>hrdB</i> as a reference. Error bars, standard deviations (<i>n</i> = 3). ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. (<b>B</b>) ChIP assays. Anti-SAV576 antibody was used to immunoprecipitate SAV576-DNA complexes from ATCC31267 and D576 cells treated with formaldehyde. The DNAs used for PCR were total DNA prior to immunoprecipitation (positive control: lanes “+”), immunoprecipitated DNA with anti-SAV576 antibody (experimental sample: lanes “S”), and immunoprecipitated DNA with rabbit preimmune serum (negative control: lanes “IgG”). The <i>hrdB</i> promoter region was used as a control. (<b>C</b>) EMSAs of the interaction of probe 577p with purified His<sub>6</sub>-SAV576 protein. Each lane contained 0.3 nM labeled probe. An approximately 100-fold excess of the unlabeled probe was used in competitive assays. BSA was used as a negative control for SAV576 protein. Labeled 262-bp <i>hrdB</i> promoter region (probe <i>hrdBp</i>) was used to eliminate non-specific binding of the SAV576 protein. The free probe is indicated by solid arrow, and the retarded DNA fragment is indicated by parentheses.</p
Pinpoint Diagnostic Kit for Heat Stroke by Monitoring Lysosomal pH
Heat
stroke is one of the most serious causes of mortality. To
prevent the situation, it is fundamental to research the mechanism
of heat cytotoxicity. The preliminary results revealed that heat stroke
and the change of lysosome acidity had some certain correlation. To
further clarify their relationship, herein, we report a highly selective
and sensitive fluorescence probe (<b>NT1</b>) for turn-on sensing
of the pH value. <b>NT2</b> was synthesized as control compound.
Compared to <b>NT2</b>, <b>NT1</b> showed accurate lysosome
target ability. In addition, the suitable p<i>K</i><sub>a</sub> value (5.67) allows <b>NT1</b> to response to the changes
of lysosomal pH values. Most importantly, <b>NT1</b> could be
used to study the correlation between the change of lysosomal pH and
heat stroke. It was shown that the lysosomal pH value increasing with
temperature during heat stroke. Thus, <b>NT1</b> was an excellent
candidate for research of the complex biological mechanism of heat
stroke
The possible regulatory pathway of SAV576/SAV577/SAV575 on avermectin biosynthesis in <i>S. avermitilis</i>.
<p>Two similar TFRs SAV576 and SAV577 both repress avermectin biosynthesis indirectly by (i) repressing <i>SAV575</i> expression directly, and SAV575 may provide precursors for avermectin biosynthesis; (ii) controlling unknown regulatory gene(s) that directly (or indirectly) regulate expression of the pathway-specific activator gene <i>aveR</i>, and AveR activates avermectin biosynthesis by activating the transcription of biosynthetic structural genes. SAV576 and SAV577 repress each other's expression. However, SAV576 is autoregulated whereas SAV577 is not. Arrows indicate activation, and bars indicate repression. Solid lines indicate direct control, and dotted lines indicate unknown routes.</p
SAV577 directly represses <i>SAV575</i> and <i>SAV576</i> expression.
<p>(<b>A</b>) Effect of <i>SAV577</i> deletion on expression of <i>aveR</i>, <i>aveA1</i>, <i>SAV575</i>, <i>SAV576</i> and <i>SAV577</i>, and detection of <i>SAV575</i> transcription level in various <i>S. avermitilis</i> strains. <i>SAV577′</i>, 136-bp transcript from <i>SAV577</i> promoter region and the remainder ORF in D577. Error bars, standard deviations (<i>n</i> = 3). ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. ns, not significant. (<b>B</b>) Schematic representation of the relative positions of probes used for EMSAs. The lengths and positions of probes 1–7 were described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099224#pone.0099224-Guo1" target="_blank">[20]</a>. Probe 577p, a 209-bp DNA fragment from positions −159 to −367 relative to the <i>SAV577</i> start codon, covering the putative transcriptional start point of <i>SAV577</i>. (<b>C</b>) EMSAs of the interaction of probes with purified His<sub>6</sub>-SAV577 protein. Each lane contained 0.3 nM labeled probe. (<b>D</b> and <b>E</b>) DNase I footprinting assay of SAV577 on the <i>SAV575</i> (<b>D</b>) and <i>SAV576</i> (<b>E</b>) promoter regions. The fluorograms represent control DNA (10 µM BSA) and protection reactions with increasing concentrations (0.08, 0.4, 0.8 µM) of His<sub>6</sub>-SAV577 protein.</p
Avermectin production and growth of wild-type strain ATCC31267 and <i>SAV577</i> mutant strains.
<p>(<b>A</b>) Comparison of avermectin production in various <i>S. avermitilis</i> strains grown in FM-I medium for 10 days. WT, wild-type strain ATCC31267. WT/pKC1139, ATCC31267 carrying control plasmid pKC1139. D576, <i>SAV576</i> deletion mutant. D577, <i>SAV577</i> deletion mutant. D577/pSET152-577, complementation strain of D577. WT/pKC1139-577, <i>SAV577</i> overexpression strain. D576-577, <i>SAV576-SAV577</i> double deletion mutant. (<b>B</b> and <b>C</b>) Effect of <i>SAV577</i> deletion on avermectin production (<b>B</b>) and growth (<b>C</b>) of <i>S. avermitilis</i> grown in soluble medium FM-II. Solid squares, ATCC31267; Solid circles, D577. Results are shown as mean ± SD for three independent experiments. **<i>P</i><0.01 and ***<i>P</i><0.001 as determined by Student's <i>t</i>-test. ns, not significant.</p
Crocin exerts anti-inflammatory and anti-arthritic effects on type II collagen-induced arthritis in rats
<p><b>Context:</b> Rheumatoid arthritis (RA) is a common systemic auto-immune disease, which is characterized by chronic and symmetry synovial inflammation. Crocin has been reported to exhibit anti-inflammatory effects in animal models.</p> <p><b>Objective:</b> This study investigates the anti-inflammatory and anti-arthritic effects of crocin on type II collagen-induced arthritis (CIA) in Wistar rats.</p> <p><b>Materials and methods:</b> The CIA rat model was established and randomly divided into five groups with or without crocin treatment (10, 20 or 40 mg/kg), which was started on day 21 after arthritis induction and persisted for 36 days. The symptoms and molecular mechanisms of CIA and crocin-treated CIA rats were compared and investigated.</p> <p><b>Results:</b> CIA rats presented severe RA symptoms, including high arthritis score, paw swelling, joint inflammation, bone erosion, chondrocyte death, cartilage destruction, enhanced expressions of matrix metalloproteinase (MMP) and pro-inflammatory cytokines. However, crocin could mitigate these symptoms. Crocin (40 mg/kg) exhibited the most efficient therapeutic function on CIA rats: the histological scores of joint inflammation, bone erosion, chondrocyte death, cartilage surface erosion, and bone erosion of CIA rats receiving 40 mg/kg crocin treatment were comparable to the normal rats. MMP-1, -3 and -13 protein expression levels of CIA rats with 40 mg/kg crocin treatment were decreased to levels similar to normal rats. Moreover, crocin could also inhibit the expression of TNF-α, IL-17, IL-6 and CXCL8 in serum and ankle tissues of CIA rats.</p> <p><b>Conclusions:</b> In summary, crocin exhibits therapeutic potential for RA, by mitigating the symptoms and inhibiting the pro-inflammatory factor expression.</p
Dentritic CuPtPd Catalyst for Enhanced Electrochemical Oxidation of Methanol
The
CuPtPd catalyst is designed and synthesized successfully via
directly reducing metal ions. The composition, morphology, and structure
of the as-prepared CuPtPd nanohybrids are characterized by scanning
electron microscopy and transmission electron microscopy, energy-dispersive
spectrometry, selected-area electron diffraction, X-ray diffraction,
and inductively coupled plasma atomic emission spectrometry. By comparison
of the electrocatalytic properties of the ternary CuPtPd catalyst
with bimetallic catalysts, we find that ternary nanocomposites perform
better electrocatalytic and antipoisoning activity toward oxidation
of methanol. The catalytic mass activity of the CuPtPd nanoparticles
is 5.51-fold of commercial Pd black and 12.1-fold of Pt black
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