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Biodegradable magnesium alloys are advantageous in various implant applications, as they reduce the risks associated with permanent metallic implants. However, a rapid corrosion rate is usually a hindrance in biomedical applications. Here we report a facile two step procedure to introduce multifunctional, anti-corrosive coatings on Mg alloys, such as AZ31. The first step involves treating the NaOH-activated Mg with bistriethoxysilylethane to immobilize a layer of densely crosslinked silane coating with good corrosion resistance; the second step is to impart amine functionality to the surface by treating the modified Mg with 3-amino-propyltrimethoxysilane. We characterized the two-layer anticorrosive coating of Mg alloy AZ31 by Fourier transform infrared spectroscopy, static contact angle measurement and optical profilometry, potentiodynamic polarization and AC impedance measurements. Furthermore, heparin was covalently conjugated onto the silane-treated AZ31 to render the coating haemocompatible, as demonstrated by reduced platelet adhesion on the heparinized surface. The method reported here is also applicable to the preparation of other types of biofunctional, anti-corrosive coatings and thus of significant interest in biodegradable implant applications.

Riboswitches are cis-acting mRNA elements that regulate gene expression in response to ligand binding. Recently, a class of riboswitches was proposed to respond to the molybdenum cofactor (Moco), which serves as a redox center for metabolic enzymes. The 5′ leader of the Escherichia coli moaABCDE transcript exemplifies this candidate riboswitch class. This mRNA encodes enzymes for Moco biosynthesis, and moaA expression is feedback inhibited by Moco. Previous RNA-seq analyses showed that moaA mRNA copurified with the RNA binding protein CsrA (carbon storage regulator), suggesting that CsrA binds to this RNA in vivo. Among its global regulatory roles, CsrA represses stationary phase metabolism and activates central carbon metabolism. Here, we used gel mobility shift analysis to determine that CsrA binds specifically and with high affinity to the moaA 5′ mRNA leader. Northern blotting and studies with a series of chromosomal lacZ reporter fusions showed that CsrA posttranscriptionally activates moaA expression without altering moaA mRNA levels, indicative of translation control. Deletion analyses, nucleotide replacement studies and footprinting with CsrA-FeBABE identified two sites for CsrA binding. Toeprinting assays suggested that CsrA binding causes changes in moaA RNA structure. We propose that the moaA mRNA leader forms an aptamer, which serves as a target of posttranscriptional regulation by at least two different factors, Moco and the protein CsrA. While we are not aware of similar dual posttranscriptional regulatory mechanisms, additional examples are likely to emerge.

Pt on Pd nanocatalyst with hollow interior structure was synthesized by a modified galvanic replacement method. The hollow nanoparticles with rough Pt surface on Pd layer (Pt–Pd) exhibit significantly higher electrochemically active surface area when compared to that of conventional hollow Pt nanoparticles, with remarkably enhanced oxygen reduction reaction activity and durability compared to those of as-synthesized hollow Pt catalysts and the E-TEK commercial Pt/C. Real-world application of the catalysts in a single fuel cell has been evaluated and found consistent with the electrochemical measurements. Keywords: Pt–Pd, Bimetallic, Electrocatalyst, Fuel cell

Graphene based materials coupled with transition metal oxides are promising electrode materials in asymmetric supercapacitors owing to their unique properties which include high surface area, good chemical stability, electrical conductivity, abundance, and lower cost profile over time. In this paper a composite material consisting of graphene oxide exfoliated with microwave radiation (mw rGO), and manganosite (MnO) is synthesised in order to explore their potential as an electrode material. The composite material was characterised by scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to explore the process occurring at the electrode/electrolyte interface. Long term cyclability and stability were investigated using galvanostatic charge/discharge testing. From the resulting analysis, an asymmetric supercapacitor was constructed with the best composite containing 90% MnO–10% mw rGO (w/w). The device exhibited a capacitance of 0.11 F/cm 2 (51.5 F/g by mass) and excellent capacity retention of 82% after 15,000 cycles at a current density of 0.5 A/g.

Noncoding regulatory RNA molecules, also known as small RNAs, participate in several bacterial regulatory networks. The central component of the carbon storage regulator (Csr) and the homologous repressor of secondary metabolites (Rsm) systems is an RNA binding protein (CsrA or RsmA) that regulates gene expression post-transcriptionally by affecting ribosome binding and/or mRNA stability. Members of the CsrB family of noncoding regulatory RNA molecules contain multiple CsrA binding sites and function as CsrA antagonists by sequestering this protein. Depending on the particular organism, the Csr (or Rsm) system participates in global regulatory circuits that control central carbon flux, the production of extracellular products, cell motility, biofilm formation, quorum sensing and/or pathogenesis.

Fungal allergens are present in viable and non-viable conidia, hyphae and fungal fragments. It has been shown that large quantities of allergen are released from conidia during germination. We used a gold immunolabelling technique and field emission scanning electron microscopy to examine the allergen release from Alternaria alternata conidia. Immunolabelling was associated with the hyphal tip and amorphous matter associated with the emerging hyphae. Non-specific antibody controls showed no labelling associated with germinating fungi. This suggests that material released from hyphae may be an additional source of fungal allergens.

The RNA-binding protein CsrA (carbon storage regulator) is a new kind of global regulator, which facilitates specific mRNA decay. A recombinant CsrA protein containing a metal-binding affinity tag (CsrA-H6) was purified to homogeneity and authenticated by N-terminal sequencing, matrix-assisted laser desorption/ionization time of flight mass spectrometry, and other studies. This protein was entirely contained within a globular complex of approximately 18 CsrA-H6 subunits and a single approximately 350-nucleotide RNA, CsrB. cDNA cloning and nucleotide sequencing revealed that the csrB gene is located downstream from syd in the 64-min region of the Escherichia coli K-12 genome and contains no open reading frames. The purified CsrA-CsrB ribonucleoprotein complex was active in regulating glg (glycogen biosynthesis) gene expression in vitro, as was the RNA-free form of the CsrA protein. Overexpression of csrB enhanced glycogen accumulation in E. coli, a stationary phase process that is repressed by CsrA. Thus, CsrB RNA is a second component of the Csr system, which binds to CsrA and antagonizes its effects on gene expression. A model for regulatory interactions in Csr is presented, which also explains previous observations on the homologous system in Erwinia carotovora. A highly repeated nucleotide sequence located within predicted stem-loops and other single-stranded regions of CsrB, CAGGA(U/A/C)G, is a plausible CsrA-binding element.

The extracellular alginate lyase activity from a fermentative marine bacterium isolated from actively growing tissues of Sargassum fluitans has been purified and studied with respect to substrate specificity and mechanism. The enzyme endolytically depolymerizes (1→4)-β- d -mannuronan derived from alginate to oligomeric products possessing 4,5-unsaturated, nonreducing termini. Reversed-phase liquid chromatography has established that early in the reaction the tri-, tetra-, and pentameric oligomers are the predominant species. The pentamer and larger products that at first accumulate in the reaction are later degraded to smaller products. The trimer is the major product late in the reaction, at which time the dimer and tetramer are also present in significant amounts. By incubating purified oligomers with enzyme, the trimer is shown to be completely refractory to further depolymerization and therefore represents a limit product of the reaction catalyzed by this enzyme. The tetramer is slowly converted into trimer and monomer, whereas the pentamer is readily converted into trimer and dimer.

The nucleotide sequences of the Escherichia coli genome between the glycogen biosynthetic genes glgB and glgC, and 1170 bp of DNA which follows glgA have been determined. The region between glgB and glgC contains an open reading frame (ORF) of 1521 bp which we call glgX. This ORF is capable of coding for an Mr 56 684 protein. The deduced amino acid (aa) sequence for the putative product shows significant similarity to the E. coli glycogen branching enzyme, and to several different glucan hydrolases and transferases. The regions of sequence similarity include residues which have been reported to be involved in substrate binding and catalysis by taka-amylase. This suggests that the proposed product may catalyze hydrolysis or glycosyltransferase reactions. The cloned region which follows glgA contains an incomplete ORF (1149 bp), glgY, which appears to encode 383 aa of the N terminus of glycogen phosphorylase, based upon sequence similarity with the enzyme from rabbit muscle (47% identical aa residues) and with maltodextrin phosphorylase from E. coli (37% identical aa residues). Results suggest that neither ORF is required for glycogen biosynthesis. The localization of glycogen biosynthetic and degradative genes together in a cluster may facilitate the regulation of these systems in vivo.

The capacities of developing Amanita carpophores to translocate and metabolically alter amatoxins were examined. The tritium-labeled derivative [ 3 H]6′- O -methyl-dehydroxymethyl-α-amanitin was administered either alone or in solution with α-amanitin or 6′- O -methyl-α-amanitin to the volvae or bulbs of immature carpophores of several nontoxic species of Amanita . Extracts of subsections of developed carpophores were analyzed by liquid scintillation counting and thin-layer chromatography/fluorography to determine translocation and chemical alterations of the [ 3 H]6′- O -methyl-dehydroxymethyl-α-amanitin. RNA polymerase II inhibition and high-performance liquid chromatography were used to quantitate the levels of amatoxins. The results of these experiments demonstrated that (a) amatoxins were translocated to the stipe and pileus in all species examined, (b) [ 3 H]6′- O -methyl-dehydroxymethyl-α-amanitin and 6′- O -methyl-α-amanitin remained unaltered after translocation, and (c) five of six nontoxic species inactivated α-amanitin. The potential for inactivation is correlated with the presence of tyrosinase activities (EC 1.14.18.1) in the carpophore tissue. It is concluded that although amatoxins are refractory toward any endopeptidolytic activities present during carpophore development, most nontoxic species produce oxidative enzymes capable of inactivating amatoxins via attack at the 6′-hydroxyindole moiety.