Almond allergens: molecular characterization, detection, and clinical relevance

Almond ( Prunus dulcis ) has been widely used in all sorts of food products (bakery, pastry, snacks), mostly due to its pleasant flavor and health benefits. However, it is also classified as a potential allergenic seed known to be responsible for triggering several mild to life-threatening immune reactions in sensitized and allergic individuals. Presently, eight groups of allergenic proteins have been identified and characterized in almond, namely, PR-10 (Pru du 1), TLP (Pru du 2), prolamins (Pru du 2S albumin, Pru du 3), profilins (Pru du 4), 60sRP (Pru du 5), and cupin (Pru du 6, Pru du γ-conglutin), although only a few of them have been tested for reactivity with almond-allergic sera. To protect sensitized individuals, labeling regulations have been implemented for foods containing potential allergenic ingredients, impelling the development of adequate analytical methods. This work aims to present an updated and critical overview of the molecular characterization and clinical relevance of almond allergens, as well as review the main methodologies used to detect and quantitate food allergens with special emphasis on almond.     

Manufacture and properties of bifidogenic saccharides derived from wood mannan

Pinus pinaster wood samples were subjected to double hydrothermal processing. The liquors coming from the second stage, containing soluble saccharides of polymeric or oligomeric nature from hemicelluloses (POHs), were subjected to membrane processing (operating in discontinuous diafiltration) for refining and fractionation. Refined POH fractions were characterized by matrix-assisted laser desorption/ionization time of flight mass spectrometry and chromatographic techniques. The most complex POH component was made up of 14 hexoses and contained 4 acetyl groups. The fermentability of purified POHs by human fecal inocula was assessed by measuring both carbon source consumption and formation of short-chain fatty acids. The bifidogenic ability of POHs was confirmed by fluorescence in situ hybridization. The stimulatory effects on the bifidobacterial population reached by POHs were of the same order as those obtained with commercial fructooligosaccharides.     

Food-derived peptides stimulate mucin secretion and gene expression in intestinal cells

In this study, the hypothesis that food-derived opioid peptides besides β-casomorphin 7 might modulate the production of mucin via a direct action on epithelial goblet cells was investigated in HT29-MTX cells used as a model of human colonic epithelium. Seven milk whey or casein peptides, a human milk peptide, and a wheat gluten-derived peptide with proved or probable ability to bind μ- or δ-opioid receptors were tested on the cell culture. Significantly increased secretion of mucins was found after exposure to six of the assayed peptides, besides the previously described β-casomorphin 7, as measured by an enzyme-linked lectin assay (ELLA). Human β-casomorphin 5 and α-lactorphin were selected to study the expression of mucin 5AC gene (MUC5AC), the HT29-MTX major secreted mucin gene. α-Lactorphin showed increased expression of MUC5AC from 4 to 24 h (up to 1.6-fold over basal level expression), although differences were statistically different only after 24 h of exposure. However, this increased expression of MUC5AC did not reach significance after cell treatment with human β-casomorphin 5. In conclusion, six food-derived peptides have been identifed with described or probable opioid activity that induce mucin secretion in HT29-MTX cells. Concretely, α-lactorphin is able to up-regulate the expression of the major secreted mucin gene encoded by these cells.     

VARIATION IN SOIL ORGANIC CARBON AND NITROGEN STOCKS ALONG A TOPOSEQUENCE IN A TRADITIONAL MEDITERRANEAN OLIVE GROVE

The impact of the topographical position on soil properties was evaluated in an olive grove with traditional tillage. Three topographical positions: summit, backslope and toeslope were chosen for evaluation. The soil samples were taken from four soil sections of 0·25 m (0–1 m). The soil organic carbon (SOC) and N content increased along the downslope direction (5·5, 6·5 and 7·1 g C kg⁻¹ and 0·3, 0·8 and 0·9 g N kg⁻¹ in the surface layer in the summit, backslope and toeslope respectively) as well as SOC and N stocks, considering the two first soil sections. In addition, there was movement of the most erodible textural fraction (silt). However, the total SOC stock (refer to 1 m of depth) did not vary with respect to the topographical position, but the total N stock (refer to 1 m of depth) varied significantly. These increases were due to erosion processes that occur along the toposequence, leading to organic matter transfers from the summit to the toeslope. All the stratification ratios calculated were lower than 2, indicating the low quality of the soils. Therefore, alternative management techniques that avoid soil erosion must be considered in olive grove in order to increase the soil quality and fertility. Copyright © 2014 John Wiley & Sons, Ltd.     

Avian pathogenic Escherichia coli MT78 invades chicken fibroblasts

Avian pathogenic Escherichia coli (APEC) are responsible for extraintestinal diseases, called colibacillosis, in avian species. The most severe manifestation of the disease is colisepticemia that usually starts at the respiratory tract and may result in bird death. However, it is not yet clear how APEC cross the respiratory epithelium and get into the bloodstream. In this work, we studied the interaction between 8 APEC strains (UEL31, UEL17, UEL13, UEL29, MT78, IMT5155, IMT2470, A2363) and a chicken non-phagocytic cell, the fibroblast CEC-32 cell line. We investigated the association profile, the invasion capability, the cytotoxicity effect and the induction of caspase-3/7 activation in an attempt to understand the way the pathogen gains access to the host bloodstream. Association to cells was determined after 1 h of infection, while cell invasion was determined after 4 and 24 h of infection. The cytotoxic effect of bacterial infection was measured by lactate dehydrogenase (LDH) release and the activation of the apoptotic program was verified by caspase-3/7 activation. Also, the presence of genes for adhesins, invasins and other related virulence-associated factors was verified by PCR. All bacterial strains showed similarity in relation to adhesion, LDH release and caspase-3/7 activation. However, one APEC strain, MT78, showed high invasion capability, comparable to the invasive Salmonella typhimurium strain SL1344. Since an APEC strain was capable of invading non-phagocytic cells in vitro, the same may be happening with the epithelial cells of the avian respiratory tract in vivo. CEC-32 monolayers can also provide a useful experimental model to study the molecular mechanisms used by APEC to invade non-phagocytic cells.     

Spatial evaluation of soil salinity using the WET sensor in the irrigated area of the Segura river lowland

The electrical conductivity of the water within the soil pores (EC_p) measured with the WET sensor, appears to be a reliable estimate of soil salinity. A methodology combining the use of the WET sensor along with geostatistics was developed to delimit and evaluate soil salinity within an irrigated area under arid to semiarid Mediterranean climate in SE Spain. A systematic random sampling of 104 points was carried out. The association between EC_p and the saturation‐extract electrical conductivity (EC_se) was assessed by means of correlation analysis. The semivariograms for EC_p were obtained at three different soil depths. Interpolation techniques, such as ordinary kriging and cokriging, were applied to obtain EC_p levels in the unknown places. For each one of the soil depths, a model able to predict EC_se from EC_p was developed by means of ordinary least squares regression analysis. A good correlation (r = 0.818, p < 0.001) between EC_p and EC_se was found. Spherical spatial distribution was the best model to fit to experimental semivariograms of EC_p at 10, 30, and 50 cm soil depths. Nevertheless, cokriging using the EC_p of an adjacent soil depth as an auxiliary variable provided the best results, compared to ordinary kriging. An analytical propagation‐error methodology was found to be useful to ascertain the contribution of the spatial interpolation and ordinary least squares analysis to the uncertainty of the EC_se mapping. This methodology allowed us to identify 98% of the study area as affected by salinity problems within a rooting depth of 50 cm, with the threshold of EC_se value at 2 dS m^–1. However, considering the crops actually grown and 10% potential reduction yield, the soil‐salinity‐affected area decreased to 83%. The use of sensors to measure soil salinity in combination with geostatistics is a cost‐effective way to draw maps of soil salinity at regional scale. This methodology is applicable to other agricultural irrigated areas under risk of salinization.     

Simultaneous determination of 20 pharmacologically active substances in cow's milk, goat's milk, and human breast milk by gas chromatography-mass spectrometry

This paper reports a systematic approach to the development of a method that combines continuous solid-phase extraction and gas chromatography-mass spectrometry for the simultaneous determination of 20 pharmacologically active substances including antibacterials (chloramphenicol, florfenicol, pyrimethamine, thiamphenicol), nonsteroideal anti-inflammatories (diclofenac, flunixin, ibuprofen, ketoprofen, naproxen, mefenamic acid, niflumic acid, phenylbutazone), antiseptic (triclosan), antiepileptic (carbamazepine), lipid regulator (clofibric acid), β-blockers (metoprolol, propranolol), and hormones (17α-ethinylestradiol, estrone, 17β-estradiol) in milk samples. The sample preparation procedure involves deproteination of the milk, followed by sample enrichment and cleanup by continuous solid-phase extraction. The proposed method provides a linear response over the range of 0.6-5000 ng/kg and features limits of detection from 0.2 to 1.2 ng/kg depending on the particular analyte. The method was successfully applied to the determination of pharmacologically active substance residues in food samples including whole, raw, half-skim, skim, and powdered milk from different sources (cow, goat, and human breast).