Transamidation Down-Regulates Intestinal Immunity of Recombinant α-Gliadin in HLA-DQ8 Transgenic Mice

Enzymatic transamidation of gliadins by microbial transglutaminase (mTG) inhibits interferon-γ (IFN-γ) secretion by intestinal T cell lines in patients with celiac disease (CD). To gain insight into the cellular mechanisms underlying the down-regulatory effects of transamidation, we tested a single recombinant α-gliadin (r-gliadin) harbouring two immunodominant peptides, p13 (aa. 120–139) and p23 (aa. 220–239), in HLA-DQ8 transgenic mice, a model of gluten sensitivity. Mice were intranasally immunised with r-gliadin or r-gliadin transamidated by mTG (K-r-gliadin) along with cholera toxin, and the response of mesenteric lymph node cells was analysed by cytokine multiplex assay. An in vitro challenge with r-gliadin was characterised by secretion of specific cytokines featuring both innate immunity and the Th1/Th2/Th17 pattern of the adaptive response. Notably, transamidation specifically down-regulated the Th1 response. Structural studies performed on K-r-gliadin confirmed that specific glutamine residues in p13 and p23, previously found to be deamidated by tissue transglutaminase, were also transamidated by mTG. In silico analysis, simulating p13 and p23 peptide binding to HLA-DQ8 showed that these glutamines, in the form of glutamate, could interact by means of salt bridges with peculiar amino acids of the alpha chain of HLA-DQ8, suggesting that their transamidation may influence the HLA-restricted recognition of these peptides. Thus, the structural findings provided a rationale to explain the down-regulation of the r-gliadin-specific Th1 response following transamidation.     

Survival and growth of Listeria monocytogenes and enterohemorrhagic Escherichia coli O157:H7 in minimally processed artichokes

The ability of Listeria monocytogenes and Escherichia coli O157:H7 inoculated by immersion (at 4.6 and 5.5 log CFU/ g, respectively) to survive on artichokes during various stages of preparation was determined. Peeling, cutting, and disinfecting operations (immersion in 50 ppm of a free chlorine solution at 4 degrees C for 5 min) reduced populations of L. monocytogenes and E. coli O157:H7 by only 1.6 and 0.8 log units, respectively. An organic acid rinse (0.02% citric acid and 0.2% ascorbic acid) was more effective than a tap water rinse in removing these pathogens. Given the possibility of both pathogens being present on artichokes at the packaging stage, their behavior during the storage of minimally processed artichokes was investigated. For this purpose, batches of artichokes inoculated with L. monocytogenes or E. coli O157:H7 (at 5.5 and 5.2 log CFU/g, respectively) were packaged in P-Plus film bags and stored at 4 degrees C for 16 days. During this period, the equilibrium atmosphere composition and natural background microflora (mesophiles, psychrotrophs, anaerobes, and fecal coliforms) were also analyzed. For the two studied pathogens, the inoculum did not have any effect on the final atmospheric composition (10% O2, 13% CO2) or on the survival of the natural background microflora of the artichokes. L. monocytogenes was able to survive during the entire storage period in the inoculated batches, while the E. coli O157:H7 level increased by 1.5 log units in the inoculated batch during the storage period. The modified atmosphere was unable to control the behavior of either pathogen.     

Mini soft cheese as a simple model for biochemical studies on cheese-making and ripening

A new miniature cheese model obtained under controlled microbiological conditions was proposed, characterized and tested for reproducibility. Optimal heat treatment of cheesemilk was defined, as well as maximal ripening time. Miniature cheeses were obtained with batch pasteurized milk (65 °C, 30 min) and ripened at 5 °C. Lactic and nonlactic microbial populations were monitored by plate counts. Proteolysis was assessed by nitrogen fractions, electrophoresis and liquid chromatography, and a sniffing test was applied to evaluate aroma. Coliform bacteria decreased during ripening but moulds and yeasts increased up to 10⁴ cfu/g after 60 d, which defined the end of ripening period. Starter population remained constant during all ripening (10⁹ cfu/g), while nonstarter lactic acid bacteria increased from ∼10² to 10⁴ cfu/g. Soluble nitrogen levels at pH 4.6, in trichloracetic acid (0.73 mol/l) and in phosphotungtic acid (0.009 mol/l) were 151, 67, and 10 g/1000 g of the total nitrogen, respectively, after 60 d of ripening, which are usual values for soft cheeses. Proteolytic patterns as measured by electrophoresis were also similar to those of standard cheeses, as well as the aroma of the products. Peptide profiles revealed that the areas of most peaks increased with ripening time. The proposed model showed to be suitable for the production of mini cheese specimens for laboratory testing of cultures and enzymes in similar conditions to their real environment in the food matrix.     

Tuning the Behavior of Au and Pt Catalysts for the Chemoselective Hydrogenation of Nitroaromatic Compounds

Efficient catalysts for the synthesis of functionalized anilines from the corresponding substituted nitroaromatics should combine an excellent chemoselectivity with a high hydrogenation activity. Attempts to increase the activity of the Au/TiO₂ catalyst and to improve the selectivity of traditional Pt catalysts, based on a previous understanding of the mode of action of supported gold and platinum nanoparticles, are reviewed and discussed.     

In situ bismuth-film electrode for square-wave anodic stripping voltammetric determination of tin in biodiesel

A bismuth-film electrode (BiFE) was applied in square-wave anodic stripping voltammetry (SWASV) in order to determine Sn (IV) in biodiesel samples. In situ simultaneous deposition of tin and bismuth at −1.2 V for 90 s was carried out in a supporting electrolyte containing 0.1 mol L⁻¹ acetate buffer (pH 4.5) and 1.73 mmol L⁻¹ caffeic acid as the complexing agent. A single well-defined anodic stripping peak was observed at −0.58 V for the oxidation of Sn to Sn (II), which was used as the analytical signal. The calibration curve was obtained in the concentration range of 0.17–7.83 μmol L⁻¹ with the detection limit being 0.14 μmol L⁻¹ (r = 0.9990). Repeatability and reproducibility for the measurement of the current peak were characterized by relative standard deviations of 3.6% and 4.1%, respectively, for a 5.0 μmol L⁻¹ Sn (IV) solution (n = 10). The method was validated by comparing the results obtained with those provided by application of the atomic absorption spectroscopy technique.     

Surface Characterization of Vulcanized Rubber Treated with Sulfuric Acid and its Adhesion to Polyurethane Adhesive

Modifications produced on a vulcanized styrene –butadiene rubber surface by treatment with sulfuric acid were studied and several experimental variables were considered.

The treatment of R1 rubber with sulfuric acid produced a noticeable decrease in contact angle which was mainly ascribed to an increase in surface energy due to the formation of sulfonic acid moieties and C?O bonds, and the removal of zinc stearate. The rubber surface swelled and became brittle as a result of the treatment, and when flexed microcracks were created. A rubber surface layer modification was produced with a consequent decrease in tensile strength and elongation-at-break values. The treatment enhanced the T-peel strength of R1 rubber/polyurethane adhesive joints and the locus of failure was cohesive in the rubber.

The optimum immersion time in H₂SO₄ solution was less than 1 min., and the reaction time in air was not found to be critical; the neutralization with ammonium hydroxide and the high concentration of the sulfuric acid (95 wt%) were essential to produce adequate effectiveness of the treatment.     

Properties of Polyurethane Elastomers with Different Hard/Soft Segment Ratio

Three polyurethane elastomers (PU) containing different hard/soft (h/s) segment ratios were prepared. The PUs were characterized using Gel Permeation Chromatography (GPC), DSC, Wide Angle X-ray Diffraction (WAXD). Dynamic Mechanical Thermal Analysis (DMTA), and Stress-controlled rheometry. The surface properties were evaluated from contact angle measurements. The PUs were used as raw materials for solvent-based adhesives, whose adhesion properties were measured from T-peel tests of solvent-wiped polyvinyl chloride (PVC)/polyurethane adhesive joints. The increase in the amount of h/s segment ratio affected the structure and morphology of the PUs, reducing the degree of phase separation and the extent of the secondary interactions between polymer chains. The h/s segment ratio determined the thermal, mechanical, rheological and adhesion properties of the PUs.     

Designing hypermedia tools for solving problems in mathematics

This article presents the design and preparation using hypermedia tools of an interactive CD-ROM for the active teaching and learning of diverse problem-solving strategies in Mathematics for secondary school students. The use of the CD-ROM allows the students to learn, interactively, the heuristic style of solving problems. A range of problems has been used, each of which requires different solving strategies. A complementary section for consulting the theoretical foundations for the process of solving problems and other related information is also included on the CD-ROM. This section provides both theoretical and curriculum support for teachers.     

Enzymatic Clarification of Fruit Juices by Fungal Pectin Lyase

The activity of pectic enzymes was measured in culture filtrates of four selected fungal strains. Pectin lyase (PL) activity was produced by all fungi, but pectinesterase activity was not detected in culture filtrates of the strain Penicillium expansum F16. The use of crude enzymes resulted in good clarification of apple juice as evaluated by an increase in transmittance at 660 nm. PL activities were partially purified by dye-affinity chromatography on Blue Sepharose. The enzyme was bound to the column and eluted in a single peak, free of PE activity, and not coinciding with the major amount of protein. Apple, grape, and passion fruit juices were effectively clarified by these partially purified enzymes. Methanol content was determined by gas-liquid chromatography, being undetectable in the clarified juices. These pectic enzymes demonstrate potential for use in fruit juice processing.     

A formal framework to test soft and hard deadlines in timed systems

This paper introduces a formal framework to specify and test systems presenting both soft and hard deadlines. While hard deadlines must always be met on time, soft deadlines can be sometimes met in a different time, usually greater, from the specified one. It is this characteristic (to formally definetextitsometimes) that produces several reasonable alternatives to define appropriate implementation relations, that is, relations to decide whether an implementation is correct with respect to a specification. In addition to introducing these relations, the paper also presents a formal testing framework to test implementations and provides an algorithm to derive sound and complete test suites with respect to the implementation relations previously defined. That is, an implementation conforms to a specification if and only if the implementation successfully passes all the tests belonging to the suite derived from the specification. Copyright © 2011 John Wiley & Sons, Ltd.