Numerical simulation of acoustic emission activity in reinforced concrete structures by means of finite element modelling at the macroscale

Acoustic emissions have been widely used as a means to investigate the damage state of concrete structures. While successful applications have regarded the localisation of cracks, quantification of the damage and safety margin estimation have been elusive because the main approaches are mostly based on empirical observations. In this work, a methodology for the numerical simulation of acoustic emission events in reinforced concrete structures is proposed with the aim of filling this gap. It relies on a numerical model for reinforced concrete structures at the macro-scale which simulates the mechanical cyclic behaviour of the structure. Analysis of the stress and strain states in the numerical model provides the basis for the simulation of the occurrence and quantification of the events. A simple attenuation law is then used to estimate the acoustic event intensity recorded by the sensors. Application to a four-point bending test on a reinforced concrete beam confirms the capability of the model to reproduce the data recorded during the test, including the Felicity effect and the cumulative intensity curve. This could potentially open the path to a more quantitative use of acoustic emission data for structural assessment of reinforced concrete structures, directly linking mechanical models and acoustic observations

C–H insertion as a key step to spiro-oxetanes, scaffolds for drug discovery

A new route to spiro-oxetanes, potential scaffolds for drug discovery, is described. The route is based on the selective 1,4-C–H insertion reactions of metallocarbenes, generated from simple carbonyl precursors in flow or batch mode, to give spiro-β-lactones that are rapidly converted into spiro-oxetanes. The three-dimensional and lead like-properties of spiro-oxetanes is illustrated by the conversion of the 1-oxa-7-azaspiro[3,5]nonane scaffold into a range of functionalized derivatives

Functional mapping of the interaction between TDP-43 and hnRNP A2 in vivo

Nuclear factor TDP-43 has been reported to play multiple roles in transcription, pre-mRNA splicing, mRNA stability and mRNA transport. From a structural point of view, TDP-43 is a member of the hnRNP protein family whose structure includes two RRM domains flanked by the N-terminus and C-terminal regions. Like many members of this family, the C-terminal region can interact with cellular factors and thus serve to modulate its function. Previously, we have described that TDP-43 binds to several members of the hnRNP A/B family through this region. In this work, we set up a coupled minigene/siRNA cellular system that allows us to obtain in vivo data to address the functional significance of TDP-43-recruited hnRNP complex formation. Using this method, we have finely mapped the interaction between TDP-43 and the hnRNP A2 protein to the region comprised between amino acid residues 321 and 366. Our results provide novel details of protein–protein interactions in splicing regulation. In addition, we provide further insight on TDP-43 functional properties, particularly the lack of effects, as seen with our assays, of the disease-associated mutations that fall within the TDP-43 321-366 region: Q331K, M337V and G348C

Identification and Characterization of a Novel Family of Cysteine-Rich Peptides (MgCRP-I) from Mytilus galloprovincialis

We report the identification of a novel gene family (named MgCRP-I) encoding short secreted cysteine-rich peptides in the Mediterranean mussel Mytilus galloprovincialis. These peptides display a highly conserved pre-pro region and a hypervariable mature peptide comprising six invariant cysteine residues arranged in three intramolecular disulfide bridges. Although their cysteine pattern is similar to cysteines-rich neurotoxic peptides of distantly related protostomes such as cone snails and arachnids, the different organization of the disulfide bridges observed in synthetic peptides and phylogenetic analyses revealed MgCRP-I as a novel protein family. Genome- and transcriptome-wide searches for orthologous sequences in other bivalve species indicated the unique presence of this gene family in Mytilus spp. Like many antimicrobial peptides and neurotoxins, MgCRP-I peptides are produced as pre-propeptides, usually have a net positive charge and likely derive from similar evolutionary mechanisms, that is, gene duplication and positive selection within the mature peptide region; however, synthetic MgCRP-I peptides did not display significant toxicity in cultured mammalian cells, insecticidal, antimicrobial, or antifungal activities. The functional role of MgCRP-I peptides in mussel physiology still remains puzzling

Plant-Growth Promotion and Biocontrol Properties of Three Streptomyces spp. Isolates to Control Bacterial Rice Pathogens

Bacterial Panicle Blight caused by Burkholderia glumae is a major disease of rice, which has dramatically affected rice production around the world in the last years. In this study we describe the assessment of three Streptomyces isolates as biocontrol agents for B. glumae. Additionally, the presence of other plant-growth promoting abilities and their possible beneficial effects upon their inoculation on rice plants was evaluated as an ecological analysis for their future inoculation in rice crops. Two isolates (A20 and 5.1) inhibited growth of virulent B. glumae strains, as well as a wide range of bacterial and fungal species, while a third strain (7.1) showed only antifungal activity. In vitro tests demonstrated the ability of these strains to produce siderophores, Indoleacetic acid (IAA), extracellular enzymes and solubilizing phosphate. Greenhouse experiments with two rice cultivars indicated that Streptomyces A20 is able to colonize rice plants and promote plant growth in both cultivars. Furthermore, an egfp tagged mutant was generated and colonization experiments were performed, indicating that Streptomyces A20 –GFP was strongly associated with root hairs, which may be related to the plant growth promotion observed in the gnotobiotic experiments. In order to characterize the antimicrobial compounds produced by strain A20 bacteria, mass spectrometry analyses were performed. This technique indicated that A20 produced several antimicrobial compounds with sizes below 3 kDa and three of these molecules were identified as Streptotricins D, E and F. These findings indicate the potential of Streptomyces A20 as a biocontrol inoculant to protect rice plants against bacterial diseases

Novel Protocol for the Chemical Synthesis of Crustacean Hyperglycemic Hormone Analogues — An Efficient Experimental Tool for Studying Their Functions

The crustacean Hyperglycemic Hormone (cHH) is present in many decapods in different isoforms, whose specific biological functions are still poorly understood. Here we report on the first chemical synthesis of three distinct isoforms of the cHH of Astacus leptodactylus carried out by solid phase peptide synthesis coupled to native chemical ligation. The synthetic 72 amino acid long peptide amides, containing L- or D-Phe3 and (Glp1, D-Phe3) were tested for their biological activity by means of homologous in vivo bioassays. The hyperglycemic activity of the D-isoforms was significantly higher than that of the L-isoform, while the presence of the N-terminal Glp residue had no influence on the peptide activity. The results show that the presence of D-Phe3 modifies the cHH functionality, contributing to the diversification of the hormone pool

Interaction of RGG and HTH motifs with nucleic acids: a study with rationally designed synthetic and recombinant polypeptides

The aim of this work is to provide data on the weak interactions between proteins and nucleic acids, using a sequence-specific DNA-binding protein (the recombinant 1-69 N-terminal domain of the phage 434 cl repressor, R69), and a non-specific nucleic acid binding domain (the arginine rich RGG domain of human nucleolin produced by solid phase synthesis) as in vitro model systems. Pyrene fluorescence spectroscopy and disulfide crosslinking via C-terminally attached Cys residues showed that cognate DNA acts as template for R69 dimerization even in the absence of the native C-terminal dimerization domain. Dimeric binding of R69 to DNA results in intensive pyrene excimer fluorescence resulting from the stacking of two C-terminally attached pyrene labels. The'fluorescence of the pyrene monomer (383nm) is quenched in a salt/sensitive manner by both cognate and non-cognate DNA molecules, probably due to electrostatic interactions with the DNA backbone. DNA sequences as short as the half-site are sufficient to promote dimer formation in a detectable manner. On the other hand, studies with artificial hybrid operators prove that two half sites provide complete specificity to the complex assembly. These studies suggest that sequential binding of monomers is a plausible pathway for the assembly of R69 on DNA. The RGG motif is present in many specific and non-specific nucleic acid binding proteins and its arginine residues undergo methylation by protein arginine methyltransferase type I as a part of various, not entirely known regulatory events. We used novel methods of solid phase peptide synthesis to produce methylated and non-methylated peptides spanning sequences from residues 646-706 of the human nucleolin C-terminal domain and we used them to study nucleic acid binding in vitro. Melting curve assays, double filter binding assays and circular dichroism studies showed that the non-sequence specific nucleic acid binding is sensitive to the geometry of the nucleic acid as it binds ssDNA>RNA>dsDNA. While binding strength is not substantially influenced by dimethylation, the ability of the peptides to modify the nuclecic acid structure, detected by CD spectroscopy, is substantially decreased upon methylation

DNA-mediated assembly of weakly interacting DNA-binding protein subunits: in vitro recruitment of phage 434 repressor and yeast GCN4 DNA-binding domains

The specificity of DNA-mediated protein assembly was studied in two in vitro systems, based on (i) the DNA-binding domain of bacteriophage 434 repressor cI (amino acid residues 1–69), or (ii) the DNA-binding domain of the yeast transcription factor GCN4, (amino acids 1–34) and their respective oligonucleotide cognates. In vivo, both of these peptides are part of larger protein molecules that also contain dimerization domains, and the resulting dimers recognize cognate palindromic DNA sequences that contain two half-sites of 4 bp each. The dimerization domains were not included in the peptides tested, so in solution—in the presence or absence of non-cognate DNA oligonucleotides—these molecules did not show appreciable dimerization, as determined by pyrene excimer fluorescence spectroscopy and oxidative cross-linking monitored by mass spectrometry. Oligonucleotides with only one 4 bp cognate half-site were able to initiate measurable dimerization, and two half-sites were able to select specific dimers even from a heterogeneous pool of molecules of closely related specificity (such as DNA-binding domains of the 434 repressor and their engineered mutants that mimic the binding helix of the related P22 phage repressor). The fluorescent technique allowed us to separately monitor the unspecific, ionic interaction of the peptides with DNA which produced a roughly similar signal in the case of both cognate and non-cognate oligonucleotides. But in the former case, a concomitant excimer fluorescence signal showed the formation of correctly positioned dimers. The results suggest that DNA acts as a highly specific template for the recruitment of weakly interacting protein molecules that can thus build up highly specific complexes

Acoustic Emissions analysis of a four-point bending test on a Reinforced Concrete beam

Assessment of existing structures by means of non-destructive techniques is an important research topic in modern engineering, because of its potential in estimating structural health status without introducing undesired damage. Among several recognized techniques, Acoustic Emission (AE) analysis is growing interest in the scientific community, since it can be used to detect the initiation of cracking processes, to determine cracking typology and to extract other information from the specimen. In this paper, the authors present the results of a four-point bending test on a Reinforce Concrete (RC) beam, performed at the University of Salerno. The test has been conducted according to a loading protocol characterised by several loading-unloading cycles, and both mechanical and acoustic data have been recorded. The results will be presented in terms of standard AE parameters, commonly adopted in literature, such as Felicity ratio, signal strength and cumulative signal strength, historic index and b value. It will be underlined that these parameters allow to detect interesting moments of the phenomenon (cracking onset and development) and to assess the damage of the structure