Limited proteolysis in the investigation of beta2-microglobulin amyloidogenic and fibrillar states.

Amyloid fibrils of patients treated with regular haemodialysis essentially consists of β2-microglobulin (β2-m) and its truncated species ΔN6β2-m lacking six residues at the amino terminus. The truncated fragment shows a higher propensity to self-aggregate and constitutes an excellent candidate for the analysis of a protein in the amyloidogenic conformation. The surface topology and the conformational analysis of native β2-m and the truncated ΔN6β2-m species both in the soluble and in the fibrillar forms were investigated by the limited proteolysis/mass spectrometry strategy. The conformation in solution of a further truncated mutant ΔN3β2-m lacking three residues at the N-terminus was also examined. This approach appeared particularly suited to investigate the regions that are solvent-exposed, or flexible enough to be accessible to protein-protein interactions and to describe the conformation of transient intermediates. Moreover, proteolysis experiments can also be tailored to investigate amyloid fibrils by discriminating the protein regions constituting the unaccessible core of the fibrils and those still flexible and exposed to the solvent. Although native β2-m and ΔN3β2-m shared essentially the same conformation, significative structural differences exist between the native and the ΔN6β2-m proteins in solution with major differences located at the end moiety of strand V and subsequent loop with strand VI and at both the N- and C-termini of the proteins. On the contrary, an identical distribution of preferential proteolytic sites was observed in both proteins in the fibrillar state, which was nearly superimposible to that observed for the soluble form of ΔN6β2-m. These data revealed that synthetic fibrils essentially consists of an unaccessible core comprising residues 20-87 of the β2-m protein with exposed and flexible N- and C-terminal ends. Moreover, proteolytic cleavages observed in vitro at Lys 6 and Lys 19 reproduce specific cleavages that have to take place in vivo to generate the truncated forms of β2-m occurring in natural fibrils. On the basis of these results, a molecular mechanism for fibril formation has been propose

optimization of solar integration in biomass fuelled steam plants

Abstract This paper is focused on solar-biomass integration and presents a thermodynamic analysis of solar power utilization replacing the steam bleeds of a regenerative Hirn cycle plant, biomass fuelled, in feedwater pre heating process. In solar-biomass integrated configuration an energy conversion efficiency of solar energy has been evaluated in order to compare the use of solar energy in solo and hybrid configurations. Such efficiency has been adopted as optimization parameter for the best hybrid plant configuration, varying steam pressure and regeneration parameters

A Hybrid Screen-Printed Strip for Enhanced Electroanalysis towards Lead and Cadmium in Multi-Matrices

Although heavy metals represent a major treat for ecosystem and human health, reference methods for their monitoring are characterized by time-consuming procedures, skilled personel and sophisticated equipment (e.g. ICP-MS, AAS). The development of portable solutions is required, particularly improving interventions and reducing complexity. To this regards, an electrochemical strip for the determination of lead and cadmium in clinical, environmental and food matrices have been developed. The Bismuth film-based flexible device has been optimized and it has been able to detect cadmium and lead, respectively, down to the detection limit of 1.3 and 2 ppb. The use of Whatman No.1 chromatographic paper has allowed to improve the sensitivity towards the detection of heavy metals, because of the porosity that allowed to pre-concentrate species. This led to an improvement in the sensitivity, with a detection limit of 0.3 and 0.5 ppb, respectively, to cadmium and lead, and offers the possibility to tune the sensitivity according to needs, e.g., improving the number of pre-concentration steps. Subsequently, the application of the electrochemical sensor in drinking water, mussel and blood serum was evaluated, demonstrating how these hybrid polyester-paper electrochemical strips can significantly lower the time and costs for on-site measurements, through analytical methods of simple use. The accuracy has been evaluated by comparison with ICP-MS measurements, giving satisfactory results

A tool for evaluating geothermal power exploitability and its application to Ischia, Southern Italy

The paper proposes a method to evaluate the potential for electric power production at any site of possible geothermal interest. Accounting for geological data of the reservoirs, the method allows the computation of the available electrical power of the investigated site. Electrical energy production from geothermal sources is realized through different techniques, such as single flash and double flash, dry steam, and binary ORC plants. The technique chosen to be the most productive is determined by analyzing a specific range of geofluid properties, mainly temperature and pressure. Moreover, each plant typology has a global efficiency that may be correlated to geofluid enthalpy by empiric relations available in literature. The proposed evaluation method brings together all these correlations, yielding the power availability from a geosource, once its temperature and pressure are known. The method takes as input the geofluid available flow rate, its pressure, temperature and non-condensable gas content. It defines the best plant option from these parameters, calculates its global efficiency and finally returns the actual available power. For sites of geothermic interest, such as the volcanic island of Ischia in Southern Italy, the results of the application of this new method clearly highlight the most suitable zones for power plants installations

Tuber borchii fruit body: 2-dimensional profile and protein identification

The formation of the fruit body represents the final phase of the ectomycorrhizal fungus T. borchii life cycle. Very little is known concerning the molecular and biochemical processes involved in the fructification phase. 2-DE maps of unripe and ripe ascocarps revealed different protein expression levels and the comparison of the electropherograms led to the identification of specific proteins for each developmental phase. Associating micropreparative 2-DE to microchemical approaches, such as N-terminal sequencing and 2-D gel-electrophoresis mass-spectrometry, proteins playing pivotal roles in truffle physiology were identifie

Serum protein profiling of early and advanced stage Crohn's disease

AbstractCrohn's disease (CD) represents a highly debilitating disease of difficult diagnosis and increasing incidence. Serum protein profiling of early stage Crohn's disease (ES) CD was investigated in order to improve the comprehension of the very early pathologic mechanisms and to support the difficult diagnostic procedures currently available. Inflammatory proteins and complement 3 chain C (C3c) were over-represented during ES CD, clusterin, retinol binding protein, α1-microglobulin and transthyretin were under-represented. A C3c isoform was found to be present only during ES CD. By now, lack of specific antibodies to detect isoforms made it impossible to perform alternative validation

Evolution of an insect immune barrier through horizontal gene transfer mediated by a parasitic wasp.

Genome sequencing data have recently demonstrated that eukaryote evolution has been remarkably influenced by the acquisition of a large number of genes by horizontal gene transfer (HGT) across different kingdoms. However, in depth-studies on the physiological traits conferred by these accidental DNA acquisitions are largely lacking. Here we elucidate the functional role of Sl gasmin, a gene of a symbiotic virus of a parasitic wasp that has been transferred to an ancestor of the moth species Spodoptera littoralis and domesticated. This gene is highly expressed in circulating immune cells (haemocytes) of larval stages, where its transcription is rapidly boosted by injection of microorganisms into the body cavity. RNAi silencing of Sl gasmin generates a phenotype characterized by a precocious suppression of phagocytic activity by haemocytes, which is rescued when these immune cells are incubated in plasma samples of control larvae, containing high levels of the encoded protein. Proteomic analysis demonstrates that the protein Sl gasmin is released by haemocytes into the haemolymph, where it opsonizes the invading bacteria to promote their phagocytosis, both in vitro and in vivo. Our results show that important physiological traits do not necessarily originate from evolution of pre-existing genes, but can be acquired by HGT events, through unique pathways of symbiotic evolution. These findings indicate that insects can paradoxically acquire selective advantages with the help of their natural enemies

Nutritional controlled preparation and administration of different tomato purées indicate increase of β-carotene and lycopene isoforms, and of antioxidant potential in human blood bioavailability: A pilot study

The isoforms of lycopene, carotenoids, and their derivatives including precursors of vitamin A are compounds relevant for preventing chronic degenerative diseases such as cardiovascular diseases and cancer. Tomatoes are a major source of these compounds. However, cooking and successive metabolic processes determine the bioavailability of tomatoes in human nutrition. To evaluate the effect of acute/chronic cooking procedures on the bioavailability of lycopene and carotene isoforms in human plasma, we measured the blood levels of these compounds and of the serum antioxidant potential in volunteers after a meal containing two different types of tomato sauce (rustic or strained). Using a randomized cross-over administration design, healthy volunteers were studied, and the above indicated compounds were determined by HPLC. The results indicate an increased bioavailability of the estimated compounds and of the serum antioxidant potential with both types of tomato purée and the subsequently derived sauces (the increase was greater with strained purée). This study sheds light on the content of nutrient precursors of vitamin A and other antioxidant compounds derived from tomatoes cooked with different strategies. Lastly, our study indicates that strained purée should be preferred over rustic purée

Shape oscillations of an oil drop rising in water: effect of surface contamination

Inertial shape oscillations of heptane drops rising in water are investigated experimentally. Diameters from 0.59 to 3.52 mm are considered, corresponding to a regime where the rising motion should not affect shape oscillations for pure immiscible fluids. The interface, however, turns out to be contaminated. The drag coefficient is considerably increased compared to that of a clean drop due to the well-known. Marangoni effect resulting from a gradient of surfactant concentration generated by the fluid motion along the interface. Thanks to the decomposition of the shape into spherical harmonics, the eigenfrequencies and the damping rates of oscillation modes n = 2, 3, 4 and 5 have been measured. Frequencies are not affected by contamination, while damping rates are increased by a considerable amount that depends neither on drop instantaneous velocity nor on diameter. This augmentation, however, depends on the mode number: it is maximum for mode two (multiplied by 2.4) and then relaxes towards the value of a clean drop as n increases. A previous similar investigation of a drop attached to a capillary has not revealed such an increase of the damping rates, indicating that the coupling between rising motion and surface contamination is responsible for this effect