The stress of food allergy issues in daily life.

OBJECTIVE: Food allergies are a growing health concern, but their implications for daily psychological functioning are unknown. This micro-longitudinal study investigated the daily frequency of food allergy issues and how this related to experiences of stress, mood and physical energy. DESIGN: One hundred and eight adults with physician-diagnosed food allergies completed an initial Internet survey followed by a 2-week Internet daily diary survey. MAIN OUTCOME MEASURES: The initial survey collected socio-demographic and food allergy information. The daily survey collected information about the participants' experiences of stress, mood, physical energy and food allergy issues during that day. RESULTS: Commonly experienced allergy issues included negative physical symptoms, higher food prices, anxiety about safety of food, trouble maintaining a healthy diet and anxiety/stress at social occasions. Furthermore, multilevel modelling analyses showed that stress and negative mood were significantly higher on days with more allergy issues. Older adults experienced lower positive mood and physical energy on days with more issues. CONCLUSION: This is the first study to incorporate near to real-time tracking to examine the frequency of food allergy issues and the implications for daily psychological functioning. Targeting the issues we identified could reduce stress in patients with food allergies and improve their overall quality of life.

Understanding the Needs of Food-Allergic Adults.

Food allergies are a growing issue, yet society's understanding of how individuals' lives are affected is limited. We conducted four focus groups with food-allergic adults in New Zealand to gain an in-depth understanding of the issues that impacted their quality of life. Key themes identified included allergen-free eating issues, health care system issues, costs of having a food allergy, effects on well-being, external influences (e.g., others' lack of awareness), and internal influences (personal growth and adaptation). The unmet needs of food-allergic adults led to risk taking, increased stress, and social isolation. A lack of awareness in others (including medical professionals, food service providers, and the general public) had a negative impact, suggesting that an intervention targeting awareness would be beneficial. Adaptation, particularly in terms of assertiveness and organizational skills, was important for coping, so we suggest training in these skills be made available for food-allergic patients.

The effect of holding live sea urchins (Evechinus chloroticus) in air prior to gonad removal on gonad adenine nucleotide profiles during storage at 4°C.

Sea urchin gonads are usually sold as a fresh chilled product. Thus, to evaluate the effect of live urchin's post-harvest conditions on gonad shelf-life, gonads were extracted either immediately after harvesting or after holding urchins in air at either 4 or 15°C for 144 and 72h, respectively. Gonads were subsequently washed in brine and stored at 4°C for 10days prior to adenine nucleotide (nmol/gw/w) profile determination. A decline in ATP (control: 376.16; urchins held in air: 231.58 and 245.16) and build-up of its degradation products, mainly inosine (control: 13.25; urchins held in air: 82.87 and 52.95), was observed in gonads recovered from urchins held in air. A faster increase in ATP degradation products was detected during storage of gonads recovered from urchins held in air, with final K-values (%) of 59.34 and 48.18 being significantly higher than K-values obtained from the controls (29.69, p<0.05), suggesting that post-harvest handling can negatively impact on gonad shelf-life.

Adsorption of enterobactin to metal oxides and the role of siderophores in bacterial adhesion to metals.

The potential contribution of chemical bonds formed between bacterial cells and metal surfaces during biofilm initiation has received little attention. Previous work has suggested that bacterial siderophores may play a role in bacterial adhesion to metals. It has now been shown using in situ ATR-IR spectroscopy that enterobactin, a catecholate siderophore secreted by Escherichia coli, forms covalent bonds with particle films of titanium dioxide, boehmite (AlOOH), and chromium oxide-hydroxide which model the surfaces of metals of significance in medical and industrial settings. Adsorption of enterobactin to the metal oxides occurred through the 2,3-dihydroxybenzoyl moieties, with the trilactone macrocycle having little involvement. Vibrational modes of the 2,3-dihydroxybenzoyl moiety of enterobactin, adsorbed to TiO(2), were assigned by comparing the observed IR spectra with those calculated by the density functional method. Comparison of the observed adsorbate IR spectrum with the calculated spectra of catecholate-type [H(2)NCOC(6)H(3)O(2)Ti(OH)(4)](2-) and salicylate-type [H(2)NCOC(6)H(3)O(2)HTi(OH)(4)](2-) surface complexes indicated that the catecholate type is dominant. Analysis of the spectra for enterobactin in solution and that adsorbed to TiO(2) revealed that the amide of the 2,3-dihydroxybenzoylserine group reorientates during coordination to surface Ti(IV) ions. Investigation into the pH dependence of enterobactin adsorption to TiO(2) surfaces showed that all 2,3-dihydroxybenzoyl groups are involved. Infrared absorption bands attributed to adsorbed enterobactin were also strongly evident for E. coli cells attached to TiO(2) particle films. These studies give evidence of enterobactin-metal bond formation and further suggest the generality of siderophore involvement in bacterial biofilm initiation on metal surfaces.

Distinct proteinase 3-induced cytokine patterns in Wegener´s granulomatosis, Churg-Strauss syndrome, and healthy controls.

OBJECTIVES: To analyse whether a specific cytokine pattern is elicited in response to the autoantigen proteinase 3 (PR3) in active Wegener's granulomatosis (WG). METHODS: Six-colour flow cytometry was used to analyse cytokine production and surface markers of the total CD4+ T-cell population ex vivo and in PR3-stimulated T-cell lines of patients with active PR3-ANCA-positive WG, PR3-ANCA-negative Churg-Strauss syndrome (CSS), and healthy controls (HC). RESULTS: The cytokine response of the total PB CD4+ T cell population was skewed towards distinct pro-inflammatory cytokine patterns in WG (Th1-type) and CSS (Th17, Th1-/Th2-type). Th2-type as well as Th17 cell populations including Th17/Th1, Th17/Th2 and Th22 cells were elicited in response to PR3 stimulation in WG. In contrast, CSS patients displayed a Th2-type dominated response following PR3 stimulation. CONCLUSIONS: These data suggest that the cytokine response of the total CD4+ T-cell population and PR3-specific cells is influenced by the underlying disorder.

Characterization of monolaurin resistance in Enterococcus faecalis.

There is increasing concern regarding the presence of vancomycin-resistant enterococci in domestically farmed animals, which may act as reservoirs and vehicles of transmission for drug-resistant enterococci to humans, resulting in serious infections. In order to assess the potential for the use of monolaurin as a food preservative, it is important to understand both its target and potential mechanisms of resistance. A Tn917 mutant library of Enterococcus faecalis AR01/DGVS was screened for resistance (MIC, >100 microg/ml) to monolaurin. Three mutants were identified as resistant to monolaurin and were designated DGRM2, DGRM5, and DGRM12. The gene interrupted in all three mutants was identified as traB, which encodes an E. faecalis pheromone shutdown protein and whose complementation in trans restored monolaurin sensitivity in all three mutants. DGRM2 was selected for further characterization. E. faecalis DGRM2 showed increased resistance to gentamicin and chloramphenicol (inhibitors of protein synthesis), while no difference in the MIC was observed with the cell wall-active antibiotics penicillin and vancomycin. E. faecalis AR01/DGVS and DGRM2 were shown to have similar rates (30% cell lysis after 4 h) of cell autolytic activity when activated by monolaurin. Differences in cell surface hydrophobicity were observed between the wild type and the mutant, with the cell surface of the parent strain being significantly more hydrophobic. Analysis of the cell wall structure of DGRM2 by transmission electron microscopy revealed an increase in the apparent cell wall thickness and contraction of its cytoplasm. Taken together, these results suggest that the increased resistance of DGRM2 was due to a change in cell surface hydrophobicity, consequently limiting the diffusion of monolaurin to a potential target in the cytoplasmic membrane and/or cytoplasm of E. faecalis.

Adsorption to metal oxides of the Pseudomonas aeruginosa siderophore pyoverdine and implications for bacterial biofilm formation on metals.

The initiation of biofilm formation is poorly understood, and in particular, the contribution of chemical bond formation between bacterial cells and metal surfaces has received little attention. We have previously used in situ infrared spectroscopy to show, during the initial stages of Pseudomonas aeruginosa biofilm formation, the formation of coordinate covalent bonds between titanium dioxide particle films and pyoverdine, a mixed catecholate and hydroxamate siderophore. Here we show using infrared spectroscopy that pyoverdine can also form covalent bonds with particle films of Fe2O3, CrOOH, and AlOOH. Adsorption to the metal oxides through the catechol-like 2,3-diamino-6,7-dihydroxyquinoline part of pyoverdine was most evident in the infrared spectrum of the adsorbed pyoverdine molecule. Weaker infrared absorption bands that are consistent with the hydroxamic acids of pyoverdine binding covalently to TiO2, Fe2O3, and AlOOH surfaces were also observed. The adsorption of pyoverdine to TiO2 and Fe2O3 surfaces showed a pH dependence that is indicative of the dominance of the catechol-like ligand of pyoverdine. Infrared absorption bands were also evident for pyoverdine associated with the cells of P. aeruginosa on TiO2 and Fe2O3 surfaces and were notably absent for genetically modified cells unable to synthesize or bind pyoverdine at the cell surface. These studies confirm the generality of pyoverdine-metal bond formation and suggest a wider involvement of siderophores in bacterial biofilm initiation on metals.

Interaction of Escherichia coli and soil particles in runoff.

A laboratory-scale model system was developed to investigate the transport mechanisms involved in the horizontal movement of bacteria in overland flow across saturated soils. A suspension of Escherichia coli and bromide tracer was added to the model system, and the bromide concentration and number of attached and unattached E. coli cells in the overland flow were measured over time. Analysis of the breakthrough curves indicated that the E. coli and bromide were transported together, presumably by the same mechanism. This implied that the E. coli was transported by advection with the flowing water. Overland-flow transport of E. coli could be significantly reduced if the cells were preattached to large soil particles (> 45 microm). However, when unattached cells were inoculated into the system, the E. coli appeared to attach predominantly to small particles (< 2 microm) and hence remained unattenuated during transport. These results imply that in runoff generated by saturation-excess conditions, bacteria are rapidly transported across the surface and have little opportunity to interact with the soil matrix.

Laboratory scale Clean-In-Place (CIP) studies on the effectiveness of different caustic and acid wash steps on the removal of dairy biofilms.

A laboratory scale, bench top flow system was used to partially reproduce dairy plant conditions under which biofilms form and to quantify the effectiveness of caustic and acid wash steps in reducing the number of viable bacteria attached to stainless steel (SS) surfaces. Once bacteria attached to surfaces, a standard clean-in-place (CIP) regime (water rinse, 1% sodium hydroxide at 65 degrees C for 10 min, water rinse, 1.0% nitric acid at 65 degrees C for 10 min, water rinse) did not reproducibly ensure their removal. Standard CIP effectiveness was compared to alternative cleaning chemicals such as: caustic blends (Alkazolv 48, Ultrazolv 700, Concept C20, and Reflex B165); a caustic additive (Eliminator); acid blends (Nitroplus and Nitrobrite); and sanitizer (Perform). The addition of a caustic additive, Eliminator, enhanced biofilm removal compared to the standard CIP regime and further increases in cleaning efficiency occurred when nitric acid was substituted with Nitroplus. The combination of NaOH plus Eliminator and Nitroplus achieved a 3.8 log reduction in the number of cells recovered from the stainless steel surface. The incorporation of a sanitizer step into the CIP did not appear to enhance biofilm removal. This study has shown that the effectiveness of a "standard" CIP can possibly be enhanced through the testing and use of caustic and acid blends. There are many implications of these findings, including: the development of improved cleaning regimes and improved product quality, plant performance, and economic returns.

Erosion and subsequent transport state of Escherichia coli from cowpats.

Processes by which fecal bacteria enter overland flow and their transportation state to surface waters are poorly understood, making the effectiveness of measures designed to intercept this pathway, such as vegetated buffer strips, difficult to predict. Freshly made and aged (up to 30 days) cowpats were exposed to simulated rainfall, and samples of the cowpat material and runoff were collected. Escherichia coli in the runoff samples were separated into attached (to particles) and unattached fractions, and the unattached fraction was analyzed to determine if the cells were clumped. Within cowpats, E. coli grew for 6 to 14 days, rather than following a typical logarithmic die-off curve. E. coli numbers in the runoff correlated with numbers inside the cowpat. Most of the E. coli organisms eroded from the cowpats were transported as single cells, and only a small percentage (about 8%) attached to particles. The erosion of E. coli from cowpats and the state in which the cells were transported did not vary with time within a single rainfall event or over time as the cowpats aged and dried out. These findings indicate that cowpats can remain a significant source of E. coli in overland flow for more than 30 days. As well, most of the E. coli organisms eroded from cowpats will occur as readily transportable single cells.

Morphotypic conversion in Listeria monocytogenes biofilm formation: biological significance of rough colony isolates.

Adherence to a stainless steel surface selected isolates of Listeria monocytogenes with enhanced surface colonization abilities and a change in phenotype from the common smooth colony morphology to a succession of rough colony morphotypes. Growth in broth culture of the best-adapted, surface-colonizing rough colony morphotype gave a smooth colony revertant. Comparative analysis revealed that the smooth and rough variants had similar phenotypic and biochemical characteristics (e.g., identical growth rates and tolerances to antibiotics and environmental stressors). Rough colony isolates, however, failed to coordinate motility or induce autolysis. The defect in autolysis of rough colony isolates, which involved impaired cellular localization of several peptidoglycan-degrading enzymes, including cell wall hydrolase A (CwhA), suggested a link to a secretory pathway defect. The genetic basis for the impairment was studied at the level of the accessory secretory pathway component SecA2. DNA sequencing of the secA2 gene in smooth and rough colony isolates found no mutations in the coding or promoter regions. Analysis of SecA2 expression with an integrated secA2-FLAG tag construct found the protein to be upregulated in the rough and revertant backgrounds compared to the parental smooth colony isolate. A compensatory mechanism involving the SecA2 secretion pathway components is postulated to control smooth to rough interconversion of L. monocytogenes. Such phenotypic variation may enhance the ability of this opportunistic pathogen to colonize environments as diverse as processing surfaces, food products, and animal hosts.

Thermal-Death Times of Listeria monocytogenes in Green Shell Mussels ( Perna canaliculus ) Prepared for Hot Smoking.

Thermal-death times were determined for seven strains of Listeria monocytogenes in green shell mussels ( Perna canaliculus ) that had been soaked in brine in preparation for hot smoking. Subsamples (25 g) of blended mussels were aseptically transferred into plastic pouches and inoculated with 106 cells of L. monocytogenes g-1. The inoculated material was incubated at six different temperatures and L. monocytogenes numbers were determined at five time intervals for each temperature. Temperature trials were duplicated. A best-fit line, by least squares linear regression, was fitted to the data. The correlation value (R2) was 0.96. From the best-fit line, D values at 56, 58, 59, 60 and 62°C were calculated to be 48.09, 16.25, 9.45, 5.49 and 1.85 min respectively, with a z value of 4.25°C. This study demonstrates that small changes in the core temperature of a product during hot smoking can have a profound effect upon the rate at which contaminating L. monocytogenes cells are killed. Results from this study will help to ensure that thermal regimes designed to eliminate L. monocytogenes from hot-smoked mussels are based on adequate thermal-time information.