Proteomic mapping of 4-hydroxynonenal protein modification sites by solid-phase hydrazide chemistry and mass spectrometry

The modification of proteins by the cytotoxic, reactive aldehyde 4-hydroxynonenal (HNE) is known to alter protein function and impair cellular mechanisms. In order to identify susceptible amino acid sites of HNE modification within complex biological mixtures by microcapillary liquid chromatography and linear ion trap tandem mass spectrometry, we have developed a solid-phase capture and release strategy that utilizes reversible hydrazide chemistry to enrich HNE-modified peptides. To maximize the detection of fragment ions diagnostic of HNE modification, both neutral loss-dependent acquisition of MS/MS/MS spectra and the pulsed Q dissociation operation mode were employed. When the solid-phase hydrazide enrichment strategy was applied to a yeast lysate treated with HNE, 125 distinct amino acid sites of HNE modification were mapped on 67 different proteins. The endogenous susceptibility of many of these proteins to HNE modification was demonstrated by analyzing HNE-treated yeast cell cultures with a complementary biotin hydrazide enrichment strategy. Further analysis revealed that the majority of amino acid sites susceptible to HNE modification were histidine residues, with most of these sites being flanked by basic amino acid residues, and predicted to be solvent exposed. These results demonstrate the effectiveness of this novel strategy as a general platform for proteome-scale identification of amino acid sites susceptible to HNE modification from within complex mixtures.     

Scanning and transmission electron microscopy study of the microstructural changes occurring in aluminium matrix composites reinforced with SiC particles during casting and welding: interface reactions

Processing of aluminium matrix composites (AMCs), especially those constituted by a reactive system such as Al–SiC, presents great difficulties which limit their potential applications. The interface reactivity between SiC and molten Al generates an aluminium carbide which degrades the composite properties. Scanning and transmission electron microscopes equipped with energy-dispersive X-ray spectroscopes are essential tools for determining the structure and chemistry of the Al–SiC interfaces in AMCs and changes occurring during casting and arc welding. In the present work, an aluminium–copper alloy (AA2014) reinforced with three different percentages of SiC particles was subjected to controlled remelting tests, at temperatures in the range 750–900 °C for 10 and 30 min. Arc welding tests using a tungsten intert gas with power inputs in the range 850–2000 W were also carried out. The results of these studies showed that during remelting there is preferential SiC particle consumption with formation of Al₄C₃ by interface reaction between the solid SiC particle and the molten aluminium matrix. The formation of Al₄C₃ by the same mechanism has also been detected in molten pools of arc welded composites. However, in this case there was formation of an almost continuous layer of Al₄C₃, which protects the particle against further consumption, and formation of aciculate aluminium carbide on the top weld. Both are formed by fusion and dissolution of the SiC in molten aluminium followed by reaction and precipitation of the Al₄C₃ during cooling.     

Antioxidant activity of grapefruit seed extract on vegetable oils

The antioxidant activities of a grapefruit seed extract (GFSE) which contains tocopherols, citric acid and ascorbic acid, were investigated in vegetable oils. Different amounts of GFSE in powder were dissolved in a mixture of soybean oil–sunflower oil. The oxidation was carried out in an AOM equipment at temperatures of 98·7°C, 75°C and 66·5°C. The results indicated prooxidant effects of GFSE at 97·8°C and 75°C possibly due to a blockade effect of a hydroxyl compound by tocopherol. The inhibitor reaction orders at these temperatures were 0·902 and 0·465, respectively, while at 66·5°C the oxidation rate was inverse to GFSE concentration with reaction order of −0·420. These results seem to indicate different reaction mechanisms depending on temperature conditions. The blockade effects were explained on the basis of a proposed complex between tocopherol and citric acid molecules. © 1998 SCI.     

Friction effects on the load capacity of a column and a hydraulic cylinder

Over the last years, a better comprehension of hydraulic cylinder performance is observed to be a prime objective of many owing to demanding applications. By observing these hydraulic cylinder applications, we see currently proposed models and design criteria do not take into account real factors, and further there are misalignment and boundary conditions in the end supports, mechanism interaction, dust, clearance and imperfections. Since these boundary conditions can develop progressive moments, the ideal simple supported bi-articulated configuration of the actuator is no longer valid, and virtual clamped configuration appears with friction moments that substantially modify the buckling analysis. In a previous work, misalignment effects were studied and discussed. In the present work a study of the influence of friction moments when misalignment effects are deliberately left aside is carried out. In order to separate these two phenomena (friction and misalignment) boundary conditions in a column are analysed. From this analysis, a theoretical and experimental work has been carried out for columns and an actuator characterizing the critical factors that cause the collapse. The aim of this paper is to describe the behaviour of actuators under load capacity with experimental validation when friction moment is taken into account and becomes an unknown variable. Experimental and theoretical results point out the importance and influence of the friction effects in columns and hydraulic cylinders. Then, it is recommended that knowledge of the hydraulic cylinder application to predict its load capacity owing to that direct extrapolation of results from current theories and criterions could lead to incorrect estimations.     

Dip-pen nanolithography on (bio)reactive monolayer and block-copolymer platforms: deposition of lines of single macromolecules

The application of atomic force microscopy (AFM) tip-mediated molecular transfer (dip-pen nanolithography or DPN) to fabricate nanopatterned (bio)reactive platforms based on dendrimers on reactive self-assembled monolayer (SAM) and polymer thin films is discussed. The transfer of high-molar-mass polyamidoamine (PAMAM) dendrimers (generation 5) and the rapid in situ covalent attachment of the deposited adsorbates onto reactive N-hydroxysuccinimide (NHS) terminated SAMs on gold and NHS-activated polystyrene-block-poly(tert-butyl acrylate) (PS(690)-b-PtBA(1210)) block copolymer thin films were investigated as strategies to suppress line broadening by surface diffusion in DPN. By exploiting carefully controlled environmental conditions (such as temperature and relative humidity), scan rates, and in particular the covalent attachment of the dendrimers to the reactive films, the observed line broadening and hence the lateral diffusion of dendrimers was substantially less pronounced compared to that observed with DPN of thiols on gold. By this method, high-definition patterns of dendrimers were conveniently fabricated down to 30-nm length scales. The presence of primary amino groups in the deposited dendrimers ultimately offers the possibility to anchor biochemically relevant molecules, such as proteins and polypeptides, to these nanostructured platforms for a wide range of possible applications in the life sciences and in particular for the investigation of controlled cell-surface interactions.     

Application of the Thermal Quadrupoles Method to Semitransparent Solids

In this study, the thermal quadrupoles method is extended to semitransparent layered solids. Using this method, the surface temperature of semitransparent multilayered materials is calculated as a function of the optical and thermal properties of each layer. This result eventually leads to determination of the thermal diffusivity, thermal resistance, and/or optical absorption coefficient of layered materials using photothermal techniques. The thermal quadrupoles method is applied to determine the thermal contact resistance in glass stacks.     

Analysis of Fin-Tube Joints in a Compact Heat Exchanger

This article deals with an analysis of fin-tube joints as functions of topological alterations of the joint fillet size. Based on numerical predictions of a joint topology formed by the surface tension driven reactive flow of molten metal, and subsequently verified by empirical evidence gathered through both laboratory and industrial testing, the topology alterations were identified for thermal integrity studies. Subsequently, thermal characteristics of corresponding fin-tube joints were determined in terms of two models of the thermal contact resistance. Model predictions of the fin efficiency with an altered topology of the joint zone were compared with the simulation results from a computational fluid dynamics study, and the results fit well. Numerical predictions of joint topology were devised using an in-house-developed finite-element code, and verified by the Surface Evolver code. Such prediction provided quantitative joint topology information that was needed in assessments of the joint thermal performance. Experimental data were obtained using a computer-controlled transparent hot zone with an ultra-high-purity nitrogen background atmosphere under tightly controlled conditions, and also by an analysis of the state-of-the-art manufacturing process data obtained from an industrial setting. It is demonstrated that a value of fin efficiency, assumed as recommended by traditional sizing design procedures, may drastically differ from actual values.     

Who leads research productivity growth? Guidelines for R&D policy-makers

This paper evaluates to what extent policy-makers have been able to promote the creation and consolidation of comprehensive research groups that contribute to the implementation of a successful innovation system. Malmquist productivity indices are applied in the case of the Spanish Food Technology Program, finding that a large size and a comprehensive multi-dimensional research output are the key features of the leading groups exhibiting high efficiency and productivity levels. While identifying these groups as benchmarks, we conclude that the financial grants allocated by the program, typically aimed at small-sized and partially oriented research groups, have not succeeded in reorienting them in time so as to overcome their limitations. We suggest that this methodology offers relevant conclusions to policy evaluation methods, helping policy-makers to readapt and reorient policies and their associated means, most notably resource allocation (financial schemes), to better respond to the actual needs of research groups in their search for excellence (micro-level perspective), and to adapt future policy design to the achievement of medium-long term policy objectives (meso and macro-level).     

Spatial and temporal variation in indicator microbe sampling is influential in beach management decisions

Fecal indicator microbes, such as enterococci, are often used to assess potential health risks caused by pathogens at recreational beaches. Microbe levels often vary based on collection time and sampling location. The primary goal of this study was to assess how spatial and temporal variations in sample collection, which are driven by environmental parameters, impact enterococci measurements and beach management decisions. A secondary goal was to assess whether enterococci levels can be predictive of the presence of Staphylococcus aureus, a skin pathogen. Over a ten-day period, hydrometeorologic data, hydrodynamic data, bather densities, enterococci levels, and S. aureus levels including methicillin-resistant S. aureus (MRSA) were measured in both water and sand. Samples were collected hourly for both water and sediment at knee-depth, and every 6 h for water at waist-depth, supratidal sand, intertidal sand, and waterline sand. Results showed that solar radiation, tides, and rainfall events were major environmental factors that impacted enterococci levels. S. aureus levels were associated with bathing load, but did not correlate with enterococci levels or any other measured parameters. The results imply that frequencies of advisories depend heavily upon sample collection policies due to spatial and temporal variation of enterococci levels in response to environmental parameters. Thus, sampling at different times of the day and at different depths can significantly impact beach management decisions. Additionally, the lack of correlation between S. aureus and enterococci suggests that use of fecal indicators may not accurately assess risk for some pathogens.