Regulation of cytotoxic T lymphocyte-associated molecule-4 by Src kinases

Cytotoxic T lymphocyte-associated molecule-4 (CTLA-4) is a cell surface receptor expressed on activated T cells that can inhibit T cell responses induced by activation of the TCR and CD28. Studies with phosphorylated peptides based on the CTLA-4 intracellular domain have suggested that tyrosine phosphorylation of CTLA-4 may regulate its interactions with cytoplasmic proteins that could determine its intracellular trafficking and/or signal transduction. However, the kinase(s) that phosphorylate CTLA-4 remain uncharacterized. In this report, we show that CTLA-4 can associate with the Src kinases Fyn and Lck and that transfection of Fyn or Lck, but not the unrelated kinase ZAP70, can induce tyrosine phosphorylation of CTLA-4 on residues Y201 and Y218. A similar pattern of tyrosine phosphorylation was found in pervanadate-treated Jurkat T cells stably expressing CTLA-4. Phosphorylation of CTLA-4 Y201 in Jurkat cells correlated with cell surface accumulation of CTLA-4. CTLA-4 phosphorylation induced the association of CTLA-4 with the tyrosine phosphatase SHP-2, but not with phosphatidylinositol 3-kinase. In contrast, Lck-induced phosphorylation of CD28 resulted in the recruitment of phosphatidylinositol 3-kinase, but not SHP-2. These findings suggest that phosphorylation of CD28 and CTLA-4 by Lck activates distinct intracellular signaling pathways. The association of CTLA-4 with Src kinases and with SHP-2 results in the formation of a CTLA-4 complex with the potential to regulate T cell activation.     

Control of foodborne pathogens on fresh-cut fruit by a novel strain of Pseudomonas graminis

The consumption of fresh-cut fruit has substantially risen over the last few years, leading to an increase in the number of outbreaks associated with fruit. Moreover, consumers are currently demanding wholesome, fresh-like, safe foods without added chemicals. As a response, the aim of this study was to determine if the naturally occurring microorganisms on fruit are “competitive with” or “antagonistic to” potentially encountered pathogens. Of the 97 and 107 isolates tested by co-inoculation with Escherichia coli O157:H7, Salmonella and Listeria innocua on fresh-cut apple and peach, respectively, and stored at 20 °C, seven showed a strong antagonistic capacity (more than 1-log unit reduction). One of the isolates, CPA-7, achieved the best reduction values (from 2.8 to 5.9-log units) and was the only isolate able to inhibit E. coli O157:H7 at refrigeration temperatures on both fruits. Therefore, CPA-7 was selected for further assays. Dose-response assays showed that CPA-7 should be present in at least the same amount as the pathogen to adequately reduce the numbers of the pathogen. From the results obtained in in vitro assays, competition seemed to be CPA-7's mode of action against E. coli O157:H7. The CPA-7 strain was identified as Pseudomonas graminis. Thus, the results support the potential use of CPA-7 as a bioprotective agent against foodborne pathogens in minimally processed fruit.     

Food and nutrition security discursive frames in the context of the Spanish economic crisis

The recent economic crisis in Spain has provoked a number of food and nutrition security (FNS) impacts that have fuelled a fragmented landscape of food-related debates, although these have not taken the form of a coherent and all-embracing public discussion on national food security. This paper is an inductive analysis of such debates, mostly based on media coverage, in an attempt to disentangle the different discourse frames on FNS in Spain, and the role played by the economic crisis in the shaping and underpinning of such frames. For this purpose a frame matrix was constructed, paying particular attention to the stakeholders’ governance claims in order to confront frame-specific problems. Results show how stakeholders have used elements of the crisis to underpin or reinterpret their core arguments. In addition, governance claims have concentrated on the malfunctioning of the food chain, mostly due to unequal bargaining power, and the way this could be putting at risk crucial components of the food system and future FNS. Also, the short-termism of most stakeholders’ approaches obscures the long run threats to the Spanish food system.     

Simulation procedure of high pressure vessels using the wire winding technique

One of the most common techniques for high pressure vessel design (>6000 bar) is the wire winding technique. This process consists on surrounding a cylinder with a flat wire made of high strength steel, reaching to each layer progressively compressive stresses in the inner cylinder. In this way, residual initial compressive stresses in the core of the vessel, in contact with the internal pressure, increase the fatigue life of the installation. Current design codes, as the ASME code (2007 Section VIII – Division 3) include special design requirements as a safety factor against plastic collapse. In particular, this code requires that the analysis shall be performed using numerical methods for cylindrical vessels that have an inner-outer diameters ratio greater than 1.5. In this work, a wire winding simulation methodology for the analysis of high pressure vessels is presented. The simulation procedure has been validated with analytical solutions in linear elastic cases, obtaining very good agreement. Then, the plastic collapse pressure has been obtained using the proposed procedure in order to carry out the ASME code requirements.     

Fate of Escherichia coli in apple and reduction of its growth using the postharvest biocontrol agent Candida sake CPA‐1

BACKGROUND: Generally, acidic fruits and fruit juices are considered ‘safe’ from a microbiological point of view. However, some outbreaks of foodborne illnesses have been linked to the consumption of unpasteurised cider. The aim of this work was to study the survival of Escherichia coli in apple juice, wounds and flesh and on apple surfaces at different temperatures and to determine the effect of the fungal biocontrol agent Candida sake CPA‐1 against the colonisation of apple by E. coli. RESULTS: Trials were conducted with a mixture of five strains of E. coli isolated from apples. E. coli was unable to grow in apple juice at 5, 15 and 25 °C but survived. At 10 °C and above, E. coli thrived in fresh‐cut apple and wounds. At 5 °C it survived in apple wounds after 27 days of storage and after 21 days in fresh‐cut apples. When E. coli was inoculated in apple wounds together with the yeast antagonist C. sake, its growth was reduced by approximately 1 log cfu wound^?1 at 25 °C. At 5 °C no effect of the biocontrol agent was observed. CONCLUSION: Despite the low pH of apple, a rapid increase in the bacterial population is possible if the temperature is not kept low enough. The biocontrol agent C. sake, developed to prevent fruit decay during storage, could also reduce E. coli growth in wounded apples at abusive temperatures. This would represent an additional benefit of using this biocontrol agent when applied to control postharvest diseases. Copyright © 2009 Society of Chemical Industry     

A model for determining the global solar radiation for Makurdi,Nigeria

An empirical model for determining the monthly average daily global solar radiation on a horizontal surface for Makurdi, Nigeria (Latitude 7°7′N and Longitude 8°6′E) was developed using the Angstrom–Page equation. The solar radiation (W/m²), hours of bright sunshine and cloudiness were measured hourly from 0600 H to 1800 H daily for 18 months. The constants ‘a’ and ‘b’ of the Angstrom linear type equation were determined by plotting the clearness index (H/H_o) against the possible sunshine hours (n_s/N) to obtain the line of best fit. The constant ‘a’ was obtained from the intercept of the line on the y-axis while the constant ‘b’ was obtained from the slope of the line. The developed model for determining the global horizontal solar radiation at the location was H = H_o [0.17 + 0.68(n/N)] with a coefficient of correlation of 0.78. The mean bias error and root mean square error that were used to test the performance of the constants were 0.17% and 1.22% respectively. The measured solar radiation was compared with the solar radiation predicted by the model and no significant difference was found between them using F-LSD at P ≤ 0.05.     

Antagonistic effect of Pseudomonas graminis CPA-7 against foodborne pathogens in fresh-cut apples under simulated commercial conditions

Recently, we reported that the application of the strain CPA-7 of Pseudomonas graminis, previously isolated from apple, could reduce the population of foodborne pathogens on minimally processed (MP) apples and peaches under laboratory conditions. Therefore, the objective of the present work was to find an antioxidant treatment and a packaging atmosphere condition to improve CPA-7 efficacy in reducing a cocktail of four Salmonella and five Listeria monocytogenes strains on MP apples under simulated commercial processing. The effect of CPA-7 application on apple quality and its survival to simulated gastric stress were also evaluated. Ascorbic acid (2%, w/v) and N-acetyl-l-cysteine (1%, w/v) as antioxidant treatments reduced Salmonella, L. monocytogenes and CPA-7 recovery, meanwhile no reduction was observed with NatureSeal^® AS1 (NS, 6%, w/v). The antagonistic strain was effective on NS-treated apple wedges stored at 10 °C with or without modified atmosphere packaging (MAP). Then, in a semi-commercial assay, efficacy of CPA-7 inoculated at 10⁵ and 10⁷ cfu mL⁻¹ against Salmonella and L. monocytogenes strains on MP apples with NS and MAP and stored at 5 and 10 °C was evaluated. Although high CPA-7 concentrations/populations avoided Salmonella growth at 10 °C and lowered L. monocytogenes population increases were observed at both temperatures, the effect was not instantaneous. No effect on apple quality was detected and CPA-7 did not survived to simulated gastric stress throughout storage. Therefore, CPA-7 could avoid pathogens growth on MP apples during storage when use as part of a hurdle technology in combination with disinfection techniques, low storage temperature and MAP.