Real-time risk analysis for hybrid earthquake early warning systems

Earthquake Early Warning Systems (EEWS), based on real-time prediction of ground motion or structural response measures, may play a role in reducing vulnerability and/or exposition of buildings and lifelines. In fact, recently seismologists developed efficient methods for rapid estimation of event features by means of limited information of the P-waves. Then, when an event is occurring, probabilistic distributions of magnitude and source-to-site distance are available and the prediction of the ground motion at the site, conditioned to the seismic network measures, may be performed in analogy with the Probabilistic Seismic Hazard Analysis (PSHA). Consequently the structural performance may be obtained by the Probabilistic Seismic Demand Analysis (PSDA), and used for real-time risk management purposes. However, such prediction is performed in very uncertain conditions which have to be taken into proper account to limit false and missed alarms. In the present study, real-time risk analysis for early warning purposes is discussed. The magnitude estimation is performed via the Bayesian approach, while the earthquake localization is based on the Voronoi cells. To test the procedure it was applied, by simulation, to the EEWS under development in the Campanian region (southern Italy). The results lead to the conclusion that the PSHA, conditioned to the EEWS, correctly predicts the hazard at the site and that the false/missed alarm probabilities may be controlled by set up of an appropriate decisional rule and alarm threshold

Exploring the use of helicogenic amino acids for optimising single chain relaxin-3 peptide agonists

Relaxin-3 is a highly conserved two-chain neuropeptide that acts through its endogenous receptor the Relaxin Family Peptide-3 (RXFP3) receptor. The ligand/receptor system is known to modulate several physiological processes, with changes in food intake and anxiety-levels the most well studied in rodent models. Agonist and antagonist analogues based on the native two-chain peptide are costly to synthesise and not ideal drug leads. Since RXFP3 interacting residues are found in the relaxin B-chain only, this has been the focus of analogue development. The B-chain is unstructured without the A-chain support, but in single-chain variants structure can be induced by dicarba-based helical stapling strategies. Here we investigated whether alternative helical inducing strategies also can enhance structure and activity at RXFP3. Combinations of the helix inducing α-aminoisobutyric acid (Aib) were incorporated into the sequence of the relaxin-3 B-chain. Aib residues at positions 13, 17 and 18 partially reintroduce helicity and activity of the relaxin-3 B-chain, but other positions are generally not suited for modifications. We identify Thr21 as a putative new receptor contact residue important for RXFP3 binding. Cysteine residues were also incorporated into the sequence and cross-linked with dichloroacetone or α, α'-dibromo--xylene. However, in contrast to previously reported dicarba variants, neither were found to promote structure and RXFP3 activity

Structure-activity studies of cysteine-rich α-Conotoxins that inhibit high voltage-activated calcium channels via GABA\u3csub\u3eB\u3c/sub\u3e receptor activation reveal a minimal functional motif

α-Conotoxins are disulfide-rich peptides that target nicotinic acetylcholine receptors. Recently we identified several α-conotoxins that also modulate voltage-gated calcium channels by acting as G protein-coupled GABAB receptor (GABABR) agonists. These α-conotoxins are promising drug leads for the treatment of chronic pain. To elucidate the diversity of α-conotoxins that act through this mechanism, we synthesized and characterized a set of peptides with homology to α-conotoxins known to inhibit high voltage-activated calcium channels via GABABR activation. Remarkably, all disulfide isomers of the active α-conotoxins Pu1.2 and Pn1.2, and the previously studied Vc1.1 showed similar levels of biological activity. Structure determination by NMR spectroscopy helped us identify a simplified biologically active eight residue peptide motif containing a single disulfide bond that is an excellent lead molecule for developing a new generation of analgesic peptide drugs

Development of relaxin-3 agonists and antagonists based on grafted disulfide-stabilized scaffolds

Relaxin-3 is a neuropeptide with important roles in metabolism, arousal, learning and memory. Its cognate receptor is the relaxin family peptide-3 (RXFP3) receptor. Relaxin-3 agonist and antagonist analogs have been shown to be able to modulate food intake in rodent models. The relaxin-3 B-chain is sufficient for receptor interactions, however, in the absence of a structural support, linear relaxin-3 B-chain analogs are rapidly degraded and thus unsuitable as drug leads. In this study, two different disulfide-stabilized scaffolds were used for grafting of important relaxin-3 B-chain residues to improve structure and stability. The use of both Veronica hederifolia Trypsin inhibitor (VhTI) and apamin grafting resulted in agonist and antagonist analogs with improved helicity. VhTI grafted peptides showed poor binding and low potency at RXFP3, on the other hand, apamin variants retained significant activity. These variants also showed improved half-life in serum from ~5 min to >6 h, and thus are promising RXFP3 specific pharmacological tools and drug leads for neuropharmacological diseases

Binding conformation and determinants of a single chain peptide antagonist at the relaxin-3 receptor RXFP3

The neuropeptide relaxin-3 and its receptor relaxin family peptide receptor-3 (RXFP3) play key roles in modulating behavior such as memory and learning, food intake and reward-seeking. A linear relaxin-3 antagonist (R3 B1-22R) based on a modified and truncated relaxin-3 B-chain was recently developed. R3 B1-22R is unstructured in solution, thus the binding conformation and determinants of receptor binding are unclear. Here we have designed, chemically synthesized and pharmacologically characterized more than 60 analogues of R3 B1-22R to develop an extensive understanding of its structure-activity relationships. We show that the key driver for affinity is the non-native C-terminal Arg23. Additional contributors to binding include amino acid residues that are important also for relaxin-3 binding, including Arg12, Ile15 and Ile19. Intriguingly, amino acid residues that are not exposed in native relaxin-3, including Phe14 and Ala17, also interact with RXFP3. We show that R3 B1-22R has a propensity to form a helical structure and modifications that support a helical conformation are functionally well tolerated while helix breakers such as proline residues disrupt binding. These data suggest the peptide adopts a helical conformation, like relaxin-3, upon binding to RXFP3 but that its smaller size allows it to penetrate deeper into the orthosteric binding site creating more extensive contacts with the receptor

Binding conformation and determinants of a single chain peptide antagonist at the relaxin-3 receptor RXFP3

The neuropeptide relaxin-3 and its receptor relaxin family peptide receptor-3 (RXFP3) play key roles in modulating behavior such as memory and learning, food intake and reward-seeking. A linear relaxin-3 antagonist (R3 B1-22R) based on a modified and truncated relaxin-3 B-chain was recently developed. R3 B1-22R is unstructured in solution, thus the binding conformation and determinants of receptor binding are unclear. Here we have designed, chemically synthesized and pharmacologically characterized more than 60 analogues of R3 B1-22R to develop an extensive understanding of its structure-activity relationships. We show that the key driver for affinity is the non-native C-terminal Arg23. Additional contributors to binding include amino acid residues that are important also for relaxin-3 binding, including Arg12, Ile15 and Ile19. Intriguingly, amino acid residues that are not exposed in native relaxin-3, including Phe14 and Ala17, also interact with RXFP3. We show that R3 B1-22R has a propensity to form a helical structure and modifications that support a helical conformation are functionally well tolerated while helix breakers such as proline residues disrupt binding. These data suggest the peptide adopts a helical conformation, like relaxin-3, upon binding to RXFP3 but that its smaller size allows it to penetrate deeper into the orthosteric binding site creating more extensive contacts with the receptor

Confronto fra analisi invasive e non invasive:studio di opere pittoriche della prima metà del XIX secolo in Italia

Enterocin NKR-5-3B, one of the multiple bacteriocins produced by Enterococcus faecium NKR-5-3, is a 64-amino acid novel circular bacteriocin that displays broad-spectrum antimicrobial activity. Here we report the identification, characterization, and three-dimensional nuclear magnetic resonance solution structure determination of enterocin NKR-5-3B. Enterocin NKR-5-3B is characterized by four helical segments that enclose a compact hydrophobic core, which together with its circular backbone impart high stability and structural integrity. We also report the corresponding structural gene, enkB, that encodes an 87-amino acid precursor peptide that undergoes a yet to be described enzymatic processing that involves adjacent cleavage and ligation of Leu(24) and Trp(87) to yield the mature (circular) enterocin NKR-5-3B