Hydraulic Safety Evaluation and Dynamic Investigations of Baghetto Bridge in Italy

The present study deals with the structural safety evaluation of a 50-year-old river bridge, called Baghetto Bridge, located in north Italy on the Adda River. Generally speaking, hydraulic processes are the main cause of bridge failure. Scour and hydrodynamic loads have been largely studied by the hydraulic engineering community; however, in practice, integration with structural analysis is often missing. The aim of this research is to provide a multidisciplinary procedure based on hydraulic and dynamic investigations devoted to the structural verification and monitoring of river bridges with traditional mechanical bearings. The deck-river interaction is addressed, studying the influence of debris accumulation on the bridge and performing structural verification of the bearing supports. The actions exerted on the bridge deck by the river current were estimated following the recommendations of the Italian code and making some further assumptions. In addition, dynamic investigations and FE modelling were performed. The results show (1) a relatively fast procedure that can be applied by practitioners to perform structural verification of river bridges with traditional mechanical bearings, and (2) an investigation method to evaluate temperature-frequency correlation as a reference for future inspections

Bioactive lipopeptides of Pseudomonas syringae

Structural features and biological activities of lipodepsipeptides produced by various strains of the widely spread phytopathogenic bacterium Pseudomonas syringae pv. syringae ae presented here. Emphasis is put on biotechnological and potential biomedical applications of this family of secondary metabolites. In this context, structurally related metabolites from other Pseudomonas species are also considered

Properties of Proteoliposomes Containing Fusicoccin Receptors from Maize

We have recently described a fusicoccin (FC)-sensitive system reconstituted by inserting into liposomes FC-receptors and H(+)-ATPase-enriched preparations from maize tissues. While the proteoliposomes of maize H(+)-ATPase had been already investigated, those of FC-receptors required a careful characterization before use in the dual system. In particular, the influence of the phospholipid environment on time-course, reversibility, and pH-dependence of the FC-binding reaction has been studied by comparing these properties in microsome-bound, solubilized, and liposome-entrapped receptors. Similarities and differences between the results of this investigation and those previously obtained with FC-receptors from spinach leaves suggest that functionally similar binding proteins from monocot and dicot plants have distinct structural features

Purification and Characterization of Leu-Proteinase, the Leucine Specific Serine Proteinase from Spinach (Spinacia oleracea L.) Leaves

The leucine specific serine proteinase present in the soluble fraction of leaves from Spinacia oleracea L. (called Leu-proteinase) has been purified by acetone precipitation and a combination of gel-filtration, ion exchange, and adsorption chromatography. This enzyme shows a molecular weight of 60,000 ± 3,000 daltons, an isoelectric point of 4.8 ± 0.1, and a relative electrophoretic mobility of 0.58 ± 0.03. The Leu-proteinase catalyzed hydrolysis of p-nitroanilides of N-α-substituted(-l-)amino acids as well as of chromogenic macromolecular substrates has been investigated between pH 5 and 10 at 23 ± 0.5°C and I = 0.1 molar. The enzyme activity is characterized by a bell-shaped profile with an optimum pH value around 7.5, reflecting the acid-base equilibrium of groups with pK(a) values of 6.8 ± 0.1 and 8.2 ± 0.1 (possibly the histidyl residue present at the active site of the enzyme and the N-terminus group). Among the substrates considered, N-α-benzoyl-l-leucine p-nitroanilide shows the most favorable catalytic parameters and allows to determine an enzyme concentration as low as 1 × 10(−9) molar. In agreement with the enzyme specificity, only N-α-tosyl-l-leucine chloromethyl ketone, di-isopropyl fluorophosphate and phenylmethylsulfonyl fluoride, among compounds considered specific for serine enzymes, strongly inhibit the Leu-proteinase. Accordingly, the enzyme activity is insensitive to cations, chelating agents, sulfydryl group reagents, and activators

Some Properties of a Functional Reconstituted Plasmalemma H(+)-ATPase Activated by Fusicoccin

Fusicoccin was shown to stimulate the ATP-driven, intravesicular acidification of liposomes reconstituted with crude fusicoccin receptors and the H(+)-translocating ATPase, both solubilized from maize (Zea mays L.) plasma membrane. The present paper reports optimal conditions for dual reconstitution and fusicoccin activation as well as the biochemical characterization of the effect of fusicoccin on this system. Fusicoccin stimulation of proton pumping was dependent on pH and fusicoccin concentration. Its specificity was demonstrated by the positive effect of two cotylenins that have a high affinity for fusicoccin receptors and by the negative response to 7,9-epideacetylfusicoccin, an inactive fusicoccin derivative. Kinetic measurements at different ATP concentrations showed that fusicoccin increases the V(max) of the enzyme. Fusicoccin stimulation of maize H(+)-ATPase was also maintained when receptors from maize were substituted by those from spinach (Spinacia oleracea L.)

Entrapment into liposomes of fusicoccin binding sites

Fusicoccin (FC) binding sites solubilised from microsomal fractions of spinach leaves have been entrapped into soybean lecithin liposomes with an 80% yield. The investigation of the properties of these proteoliposomes has demonstrated that the rates of FC-binding and of exchange between radioactive and cold FC are intermediate between those observed with membrane-bound and with solubilised binding sites. It appears that the entrapped proteins are preferentially outside-oriented since they are inactivated by trypsin treatment; moreover, Triton X-100 permeabilization of the proteoliposomes demonstrated that one fifth of these sites are inside-oriented