Simultaneous identification of GSTP1 Ile105→Val105 and Ala114→Val114 substitutions using an amplification refractory mutation systempolymerase chain reactionassay: studies in patients with asthma

BACKGROUND: The glutathione S-transferase (GST) enzyme GSTP1 utilizes byproducts of oxidative stress. We previously showed that alleles of GSTP1 that encode the Ile105→Val105 substitution are associated with the asthma phenotypes of atopy and bronchial hyperresponsiveness (BHR). However, a further polymorphic site (Ala114→Val114) has been identified that results in the following alleles: GSTP1(*)A (wild-type Ile105→Ala114), GSTP1(*)B (Val105→Ala114), GSTP1(*)C (Val105→Val114) and GSTP1(*)D (Ile105→Val114). METHODS: Because full identification of GSTP1 alleles may identify stronger links with asthma phenotypes, we describe an amplification refractory mutation system (ARMS) assay that allows identification of all genotypes. We explored whether the GSTP1 substitutions influence susceptibility to asthma, atopy and BHR. RESULTS: Among 191 atopic nonasthmatic, atopic asthmatic and nonatopic nonasthmatic individuals, none had the BD, CD, or DD genotypes. GSTP1 BC was significantly associated with reduced risk for atopy (P = 0.031). Compared with AA, trend test analysis identified a significant decrease in the frequency of GSTP1 BC with increasing severity of BHR (P = 0.031). Similarly, the frequency of GSTP1 AA increased with increasing BHR. CONCLUSION: These data suggest that GSTP1(*)B and possibly GSTP1(*)C are protective against asthma and related phenotypes

Perspectives for the VITO beam line at ISOLDE, CERN

By using polarized ion beams in combination with the β-NMR technique, the Versatile Ion-polarized Techniques On-line (VITO) experiment at ISOLDE, CERN links together expertise from different fields in an unique experimental setup. An overview of the experimental techniques and a general description of the newly designed beam line are presented. Potential uses in multidisciplinary research and perspectives for future experiments are discussed

Controlled exposure to diesel exhaust and traffic noise - Effects on oxidative stress and activation in mononuclear blood cells

Particulate air pollution increases risk of cancer and cardiopulmonary disease, partly through oxidative stress. Traffic-related noise increases risk of cardiovascular disease and may cause oxidative stress. In this controlled random sequence study, 18 healthy subjects were exposed for 3h to diesel exhaust (DE) at 276μg/m(3) from a passenger car or filtered air, with co-exposure to traffic noise at 48 or 75dB(A). Gene expression markers of inflammation, (interleukin-8 and tumor necrosis factor), oxidative stress (heme oxygenase (decycling-1)) and DNA repair (8-oxoguanine DNA glycosylase (OGG1)) were unaltered in peripheral blood mononuclear cells (PBMCs). No significant differences in DNA damage levels, measured by the comet assay, were observed after DE exposure, whereas exposure to high noise levels was associated with significantly increased levels of hOGG1-sensitive sites in PBMCs. Urinary levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine were unaltered. In auxiliary ex vivo experiments whole blood was incubated with particles from the exposure chamber for 3h without effects on DNA damage in PBMCs or intracellular reactive oxygen species production and expression of CD11b and CD62L adhesion molecules in leukocyte subtypes

Multi-functional oilfield production chemicals: maleic-based polymers for gas hydrate and corrosion inhibition

Several chemical problems can occur during the production of oil and gas through flow lines. This includes corrosion, scale deposition and gas hydrate plugging. Three separate chemicals may be needed to treat these issues. Kinetic hydrate inhibitors (KHIs) are used in cold oil or natural gas production flow lines to prevent the formation and plugging of the line with gas hydrates. They are often injected concomitantly with other production chemicals such as corrosion and scale inhibitors. KHIs are specific low molecular weight water-soluble polymers with amphiphilic groups formulated with synergists and solvents. However, many corrosion inhibitors (CIs) are antagonistic to the KHI polymer, severely reducing the KHI performance. It would be preferable and economic if the KHI also could act as a CI. We have explored the use of maleic-based copolymers as KHIs as well as their use as film-forming CIs. KHIs were tested using a natural gas mixture in high pressure rocking cells using the slow constant cooling test method. A terpolymer from reaction of vinyl acetate:maleic anhydride copolymer with cyclohexy lamine and 3,3-di-n-butylaminopropylamine (VA:MA-60% cHex-40% DBAPA), gave excellent performance as a KHI, better than the commercially available poly(N-vinyl caprolactam) (PVCap). CO2 corrosion inhibition was measured by Linear Polarization Resistance (LPR) in a 1 litre CO2 bubble test equipment using C1018 steel coupons. The new terpolymer gave good CO2 corrosion inhibition in 3.6 wt% brine, significantly better than PVCap, but not as good as a commercial imidazoline-based surfactant corrosion inhibitor. The terpolymer also showed good corrosion inhibition efficiency at high salinity conditions, (density 1.12 g/cm3). VA:MA-60% cHex-40% DBAPA shifted the open-circuit potential to more positive values and significantly decreased the corrosion rate.publishedVersio

Water and molecular chaperones act as weak links of protein folding networks: energy landscape and punctuated equilibrium changes point towards a game theory of proteins

Water molecules and molecular chaperones efficiently help the protein folding process. Here we describe their action in the context of the energy and topological networks of proteins. In energy terms water and chaperones were suggested to decrease the activation energy between various local energy minima smoothing the energy landscape, rescuing misfolded proteins from conformational traps and stabilizing their native structure. In kinetic terms water and chaperones may make the punctuated equilibrium of conformational changes less punctuated and help protein relaxation. Finally, water and chaperones may help the convergence of multiple energy landscapes during protein-macromolecule interactions. We also discuss the possibility of the introduction of protein games to narrow the multitude of the energy landscapes when a protein binds to another macromolecule. Both water and chaperones provide a diffuse set of rapidly fluctuating weak links (low affinity and low probability interactions), which allow the generalization of all these statements to a multitude of networks.Comment: 9 pages, 1 figur

Reduction of aerobic and lactic acid bacteria in dairy desludge using an integrated compressed CO2 and ultrasonic process

International audienceAbstractCurrent treatment routes are not suitable to reduce and stabilise bacterial content in some dairy process streams such as separator and bactofuge desludges which currently present a major emission problem faced by dairy producers. In this study, a novel method for the processing of desludge was developed. The new method, elevated pressure sonication (EPS), uses a combination of low frequency ultrasound (20 kHz) and elevated CO2 pressure (50 to 100 bar). Process conditions (pressure, sonicator power, processing time) were optimised for batch and continuous EPS processes to reduce viable numbers of aerobic and lactic acid bacteria in bactofuge desludge by ≥3-log fold. Coagulation of proteins present in the desludge also occurred, causing separation of solid (curd) and liquid (whey) fractions. The proposed process offers a 10-fold reduction in energy compared to high temperature short time (HTST) treatment of milk

Structure of metal site in azurin, met 121 mutants of azurin, and stellacyanin investigated by 111m Cd Perturbed Angular Correlation (PAC)

The geometries of the metal sites in cadmium-substituted azurins have been investigated by Cd-111m perturbed angular correlation (PAC), The study includes wild type azurin as well as Met(121) mutants of azurin, where methionine has been substituted by Ala, Asn, Asp, Gin, Glu, and Leu.The nuclear quadrupole interaction of wild type azurin analyzed in the angular overlap model is well described as coordination of His(46), His(117), and Cys(112) and cannot be described by coordination of Met(121) and/or Gly(45).For most of the mutants, there exist two coordination geometries of the cadmium ion, With the exception of the Gau and Asp mutants, one of the conformations is similar to the wild type conformation. The other coordination geometries are either best described by a coordinating water molecule close to the original methionine position or by coordination by the substituting amino acid, These experiments show that even though the methionine does not coordinate it plays an important role for the geometry of the metal site.The nuclear quadrupole interaction of stellacyanin was also measured, The value resembles the most prominent nuclear quadrupole interaction of the Met(121) --> Gin mutant of Alcaligenes denitrificans azurin, indicating that the structures of the two metal sites are similar.Macromolecular Biochemistr