Vascular proteomics

The characterization of patients with acute coronary syndromes (ACS) at the molecular and cellular levels provides a novel vision for understanding the pathological and clinical expression of the disease. Recent advances in proteomic technologies permit the evaluation of systematic changes in protein expression in many biological systems and have been extensively applied to cardiovascular diseases (CVD). The cardiovascular system is in permanent intimate contact with blood, making blood-based biomarker discovery a particularly worthwhile approach. Thus, proteomics can potentially yield novel biomarkers reflecting CVD, establish earlier detection strategies, and monitor response to therapy. Here we review the different proteomic strategies used in the study of atherosclerosis and the novel proteins differentially expressed and secreted by atherosclerotic lesions which constitute novel potential biomarkers (HSP-27, Cathepsin D). Special attention is paid to MS-Imaging of atheroma plaque and the generation, for the first time, of 2-D images of lipids, showing the distribution of these molecules in the different areas of the atherosclerotic lesions. In addition new potential biomarkers have been identified in plasma (amyloid A1α, transtherytin), circulating cells (protein profile in monocytes from ACS patients) and individual cells constituents of atheroma plaques (endothelial, VSMC, macrophages) which provide novel insights into vascular pathophysiology.     

Anaerobic monodigestion of poultry manure: determination of operational parameters for CSTR

In this work the anaerobic monodigestion for the treatment of turkey manure was evaluated, without its codigestion with another substrate. The effect of the organic loading rate (OLR) and the substrate concentration (high total solids (TS) concentration) or product concentration (high volatile fatty acids (VFA) and/or ammonia (NH(3)-N) concentrations) was studied. The results show that for a continuous stirred tank reactor (CSTR) operation, a maximum of 40 g/L of TS and 4.0 g/L of ammonium (NH(4)(+)) was required. In addition, the maximum organic loading rate (OLR) will not exceed 1.5 kg VS/m(3)d. Higher TS and NH(4)(+) concentrations and OLR lead to a reduction on the methane productivity and volatile solids (VS) removal. During the CSTR operation, a high alkalinity concentration (above 10 g/L CaCO(3)) was found; this situation allowed maintaining a constant and appropriate pH (close to 7.8), despite the VFA accumulation. In this sense, the alkalinity ratio (α) is a more appropriate control and monitoring parameter of the reactor operation compared to pH. Additionally, with this parameter a VS removal of 80% with a methane productivity of 0.50 m(3)(CH4)/m(3)(R)d is achieved.     

On drawbody shapes: From Bergmark–Roos to kinematic models

The geometrical locus defined by the initial location of the fragments that are recovered from an extraction point in underground mining, after a given operation, is commonly named “drawbody”. A brief review of drawbody shapes in flat-bottomed hoppers is proposed. The Bergmark–Roos hypothesis is discussed and it is shown that when the continuity equation is considered, particle density increases with time and when moving toward the hopper aperture. Drawbody shapes are calculated for flows predicted from a plasticity approach, as well as from a kinematic model. Applications to complex configurations in which the flow is produced by two drawpoints, either in simultaneous or sequential extractions, are discussed in some detail. In particular, the extracted zone is calculated exactly and its dependence on distance between drawpoints is investigated. The knowledge of such locus should prove valuable when optimizing ore recovery in mining processes.     

A small domain (6.5 kDa) of bacterial protein G inhibits C3 covalent binding to the Fc region of IgG immune complexes

Attachment of the complement component C3 to antigen-antibody (Ag-Ab) complexes (immune complexes, IC) is the key molecular event responsible for the elimination of many Ag in the form of Ag-Ab-C3b. The CH1 domain and the Fc region of the Ab, which have previously been involved in the binding of C3b, are also the targets of several bacterial IgG-binding proteins, particularly proteins G and A. Here we describe the ability of a small recombinant protein G domain (B2; 6.5 kDa) to inhibit the covalent binding of C3b to the Fc portion of IgG without affecting the binding to the Fab part. Protein G (B2 domain) produced a remarkable inhibition of covalent binding of C3b to IC formed with rabbit IgG, but none with the F(ab')2 fragment, indicating that B2 interferes with the C3b binding to the Fc region. A weak inhibition was observed with IC formed with mouse IgG2b which preferentially binds B2 domain on the CH1 domain of the Fab. To confirm these data, recombinant single-chain Ab devoid of CH1 domains (scAb), and including the rabbit or human Fc portion (hinge-CH2-CH3), were produced and used to form IC. Protein G-B2 domain inhibited C3b binding to IC formed with scAb of either human or rabbit constant regions, supporting the view of a specific blockade of C3b binding to the Fc region. A similar inhibition of C3b binding was observed using protein A instead of protein G B2 domain and the same set of IC. On the CH1 domain, C3b and B2 bind on opposite faces, and therefore do not interfere with each other in their binding. However, B2 domain bound to the inter-CH2-CH3 region impedes the C3b binding to the Fc. This inhibition clarifies the specificity of C3b for the different regions of IgG and explains how bacterial IgG-binding proteins provide the bacteria with a mechanism of evasion from the opsonizing action of complement and contribute to the virulence. This could be a general mechanism of escape because protein G binds the majority of mammalian Ig.     

Rancidity development during the frozen storage of farmed coho salmon (Oncorhynchus kisutch): Effect of antioxidant composition supplied in the diet

A commercial diet including synthetic antioxidants (BHT–ethoxyquin mixture) (diet I) was provided to coho salmon (Oncorhynchus kisutch) in parallel with two diets including natural antioxidants (tocopherol isomers–rich mixture, diet II; tocopherol isomers-rosemary extract mixture, diet III). A comparative study of the rancidity development in the corresponding frozen (−18 °C) products was undertaken. When compared to fish fed with diet I, individuals corresponding to diet II showed a greater (p < 0.05) retention of primary (conjugated dienes and peroxides content) and secondary (anisidine and thiobarbituric acid indices) lipid oxidation compounds that led to a lower interaction compound formation (fluorescence ratio ranges: 0.33–0.50 and 0.55–0.85, for diet II and diet I individuals, respectively); likewise, a higher polyene index (1.99–2.14 and 1.72–1.97, respectively) and lower oxidised taste scores (0.0–0.6 and 0.0–2.4, respectively) were obtained. No effect (p > 0.05) on lipid hydrolysis development (free fatty acid formation) could be found as a result of employing different diets.