Fluorophore‐Promoted RNA Folding and Photostability Enables Imaging of Single Broccoli‐Tagged mRNAs in Live Mammalian Cells

Spinach and Broccoli are fluorogenic RNA aptamers that bind DFHBI, a mimic of the chromophore in green fluorescent protein, and activate its fluorescence. Spinach/Broccoli‐DFHBI complexes exhibit high fluorescence in vitro, but they exhibit lower fluorescence in mammalian cells. Here, computational screening was used to identify BI, a DFHBI derivative that binds Broccoli with higher affinity and leads to markedly higher fluorescence in cells compared to previous ligands. BI prevents thermal unfolding of Broccoli at 37 °C, leading to more folded Broccoli and thus more fluorescent Broccoli‐BI complexes in cells. Broccoli‐BI complexes are more photostable owing to impaired photoisomerization and rapid unbinding of photoisomerized cis‐BI. These properties enable single mRNA containing 24 Broccoli aptamers to be imaged in live mammalian cells treated with BI. Small molecule ligands can thus promote RNA folding in cells, and thus allow single mRNA imaging with fluorogenic aptamers.     

An algorithm for unimodular completion over Laurent polynomial rings

We present a new and simple algorithm for completion of unimodular vectors with entries in a multivariate Laurent polynomial ring over an infinite field K. More precisely, given n?3 and a unimodular vector V=^t(v₁,…,v_n)∈Rⁿ (that is, such that 〈v₁,…,v_n〉=R), the algorithm computes a matrix M in M_n(R) whose determinant is a monomial such that MV=^t(1,0,…,0), and thus M^-1 is a completion of V to an invertible matrix.     

Improved image contrast with mangafodipir trisodium (MnDPDP) during MR-guided percutaneous cryosurgery of the liver

The present study was undertaken to measure the gain observed in the liver-to-tumor contrast of perioperative images when using mangafodipir trisodium, a liver-specific contrast agent, during percutaneous cryosurgery of the liver performed under the guidance of magnetic resonance images. Retrospective quantitative analyses of MR images were performed on eleven patients having a total of 30 liver tumors treated by MR-guided percutaneous cryosurgery. An initial group of four patients were treated with no contrast agent, and was compared with a second group of 7 patients who received an intravenous injection of 5 microM/kg of mangafodipir for their cryosurgery. The percutaneous cryosurgery was monitored under the near-real-time-imaging mode of a 0.5T open-configuration MRI system using a T(1)-weighted Gradient-recalled echo pulse sequence. A significant improvement in the liver-to-tumor contrast-to-noise ratio was observed with mangafodipir (p < 0.05, paired t test) in 0.5T preoperative images. Along with the stability of the mangafodipir contrast enhancement during the entire cryosurgical procedure, the resulting gain in contrast allowed for better visualizing the presence of residual untreated tumor margins at the periphery of the cryosurgery iceball directly from perioperative images acquired with patients under narcosis. Consequently, it not only became easier for the interventionalist to determine the need for an additional cryoprobe to increase the size of the iceball during the procedure, but also to decide on the appropriate end point of the cryosurgery.     

Nitrosopeptide mapping: a novel methodology reveals s-nitrosylation of dexras1 on a single cysteine residue

S-Nitrosylation of specific cysteine residues is a reversible signaling mechanism of nitric oxide (NO) generated by NO synthase (NOS) enzymes. In some proteins, evidence has accumulated that more than one cysteine can be S-nitrosylated; however, it is difficult to distinguish S-nitrosylation on separate cysteine residues. We report a novel simple, sensitive, and specific procedure for nitrosopeptide mapping. Dexras1 is a monomeric G protein whose guanine nucleotide exchange activity is augmented by NO; the identity and number of its S-nitrosylated cysteines is unknown. We describe the radiolabeling of S-nitrosylated cysteine residues in Dexras1. A nitrosopeptide map, generated by two-dimensional peptide chromatography, reveals that only a single cysteine is S-nitrosylated following NO exposure. Mutagenesis of Cys11 abolished the effect of NO donors on Dexras1, implicating this residue in the NO-mediated activation of Dexras1.     

Fluorophore-Promoted RNA Folding and Photostability Enables Imaging of Single Broccoli-Tagged mRNAs in Live Mammalian Cells

Spinach and Broccoli are fluorogenic RNA aptamers that bind DFHBI, a mimic of the chromophore in green fluorescent protein, and activate its fluorescence. Spinach/Broccoli-DFHBI complexes exhibit high fluorescence in vitro, but they exhibit lower fluorescence in mammalian cells. Here, computational screening was used to identify BI, a DFHBI derivative that binds Broccoli with higher affinity and leads to markedly higher fluorescence in cells compared to previous ligands. BI prevents thermal unfolding of Broccoli at 37 °C, leading to more folded Broccoli and thus more fluorescent Broccoli-BI complexes in cells. Broccoli-BI complexes are more photostable owing to impaired photoisomerization and rapid unbinding of photoisomerized cis-BI. These properties enable single mRNA containing 24 Broccoli aptamers to be imaged in live mammalian cells treated with BI. Small molecule ligands can thus promote RNA folding in cells, and thus allow single mRNA imaging with fluorogenic aptamers.     

First Report of a Lipopeptide Biosurfactant from Thermophilic Bacterium Aneurinibacillus thermoaerophilus MK01 Newly Isolated from Municipal Landfill Site

A biosurfactant-producing thermophile was isolated from the Kahrizak landfill of Tehran and identified as a bacterium belonging to the genus Aneurinibacillus. A thermostable lipopeptide-type biosurfactant was purified from the culture medium of this bacterium and showed stability in the temperature range of 20–90 °C and pH range of 5–10. The produced biosurfactant could reduce the surface tension of water from 72 to 43 mN/m with a CMC of 1.21 mg/mL. The strain growing at a temperature of 45 °C produces a substantial amount of 5 g/L of biosurfactant in the medium supplemented with sunflower oil as the sole carbon source. Response surface methodology was employed to optimize the biosurfactant production using sunflower oil, sodium nitrate, and yeast extract as variables. The optimization resulted in 6.75 g/L biosurfactant production, i.e., 35 % improved as compared to the unoptimized condition. Thin-layer chromatography, FTIR spectroscopy, ¹H-NMR spectroscopy, and biochemical composition analysis confirmed the lipopeptide structure of the biosurfactant.     

A newly synthesized organic–inorganic hybrid in nano-size including [BW12O40]5− anions and hydrolyzed 2-cyanoguanidine cations as a double working green catalyst

Research on Chemical Intermediates - A newly prepared nano-organic–inorganic hybrid compound, videlicet [HPDCD]5[BW12O40]·3H2O (HPDCD?=?hydrolyzed and protonated...     

Quantitative sodium magnetic resonance imaging in food: Addressing sensitivity issues using single quantum chemical shift imaging at high field

According to various health organizations, the global consumption of salt is higher than recommended and needs to be reduced. Ideally, this would be achieved without losing the taste of the salt itself. In order to accomplish this goal, both at the industrial and domestic levels, we need to understand the mechanisms that govern the final distribution of salt in food. The in-silico solutions in use today greatly over-simplify the real food structure. Measuring the quantity of sodium at the local level is key to understanding sodium distribution. Sodium magnetic resonance imaging (MRI), a non-destructive approach, is the ideal choice for salt mapping along transformational process. However, the low sensitivity of the sodium nucleus and its short relaxation times make this imaging difficult. In this paper, we show how sodium MRI can be used to highlight salt heterogeneities in food products, provided that the temporal decay is modeled, thus correcting for differences in relaxation speeds. We then propose an abacus which shows the relationship between the signal-to-noise ratio of the sodium MRI, the salt concentration, the B0 field, and the spatial and temporal resolutions. This abacus simplifies making the right choices when implementing sodium MRI.     

Assembling ligands in situ using bioorthogonal boronate ester synthesis

Many molecules that could manipulate cellular function are not practical due to their large size and concomitant undesirable pharmocokinetic properties. Here, we describe a bioorthogonal, highly stable boronate ester (HiSBE) synthesis and use this reaction to synthesize a biologically active molecule from smaller precursors in a physiological context. The rapid rate of HiSBE synthesis suggests that it may be useful for assembling a wide variety of biologically active molecules in physiological solutions.     

Antibacterial and antioxidant constituents of Acalypha wilkesiana

This study was aimed at characterising the secondary metabolites responsible for antibacterial and antioxidant activities of Acalypha wilkesiana. Purification of the defatted methanol leaves extract was guided by the DPPH free radical scavenging assay as well as by evaluation of the antibacterial activity against four bacterial strains. As a result, geraniin, corilagin, quadrangularic acid M and shikimic acid were purified and isolated. Shikimic acid, reported for the first time from this plant, proved to be the major metabolite of the extract. All the four isolated compounds showed bactericidal activity against extended spectrum beta-lactamase-producing Klebsiella pneumoniae (700603), while corilagin was the single compound to exhibit antioxidant activity (IC₅₀ 53 μg/mL).