R-mandelonitrile-lyase, homolog to Pru d 10, is a major peach allergen in peach allergic Spanish population.

BACKGROUND AND OBJECTIVE: Peach allergy is a prevalent cause of food allergy. Despite the repertoire of allergens available for molecular diagnosis, there are still patients with undetectable IgE levels to peach allergens but presenting symptoms after its ingestion. The objective of this study was to investigate the allergenic profile in a patient population with symptoms produced by peach. METHODS: An exploratory retrospective study was performed with patients presenting symptoms after the ingestion of peach. Forty-two patients were included in the study. The allergenic profile of individual patients was investigated by immunoblot. A serum pool was prepared with the sera that recognized a 70 kDa band. This pool was used to detect this protein in peach peel and pulp and to identify the 70 kDa protein in 2D immunoblot. Spots recognized in the 2D immunoblot were sequenced by LC-MS/MS. Inhibition studies were performed between peach peel and almond. RESULTS: Twenty-two patients (52.4%) recognized the 70 kDa protein in immunoblot. This protein was recognized in peel and pulp. Two different spots were observed in 2D-PAGE, both were identified as (R)-mandelonitrile lyases (RML) with high amino acid similarity with Pru du 10. Peach RML were partially inhibited with an almond extract. No association was found between any reported symptom and sensitization to RML. RML-sensitized patients were older and reported pollen associated respiratory symptoms more frequently than negative patients. CONCLUSION: A new peach allergen, a RML, homologous of Pru du 10, recognized by 52% of the population has been identified.

IgE, IgG1 and IgG4 response to specific allergens in sensitized subjects showing different clinical reactivity to Anisakis simplex.

SUMMARY: Background. Anisakis simplex hypersensitive subjects may be sensitized without clinical allergy, or experience acute symptoms or chronic urticaria induced by raw fish. We studied whether the 3 subgroups differ in IgE, IgG1 or IgG4 reactivity to specific Anisakis simplex allergens. Methods. 28 Anisakis simplex-hypersensitive adults, 11 with acute symptoms, 9 with chronic urticaria, and 8 sensitized were studied. IgE, IgG1 and IgG4 to rAni s 1, 5, 9 and 10 were sought by ELISA. IgE and IgG4 to nAni s 4 were determined by WB. Results. IgE to Ani s 1, 4, 5, 9, and 10 were found in 8, 3, 2, 5, and 9 sera, respectively. Nine sera did not react to any allergen. IgG1 to Ani s 1, 5, 9, and 10 were detected in 5, 16, 14, and 4 sera, respectively. Four sera did not react to any of the 4 allergens. IgG4 to Ani s 1, 4, 5, 9, and 10 were detected in 10, 0, 2, 6 and 1 sera, respectively. Fifteen subjects did not react to any of the 5 allergens. On ELISA sensitized subjects showed lower IgE and IgG1 levels than patients. IgG4 levels were highest in the sensitized group. The prevalence of IgE, IgG1 or IgG4 reactivity to any of the studied allergens did not differ between the 3 subgroups. Conclusion. The clinical expression of Anisakis simplex sensitization does not seem to depend on IgE reactivity to a specific allergen of the parasite, nor on the presence of IgG antibodies possibly related with blocking activity.

Relationships between T cell and IgE/IgG4 epitopes of the Anisakis simplex major allergen Ani s 1.

BACKGROUND: Anisakiasis is a global disease caused by the consumption of raw or lightly cooked fish parasitized with third-stage Anisakis larvae. Anisakis simplex allergens may cause severe allergic reactions including angio-oedema, urticaria and anaphylaxis. Approximately 80% of allergic patients have allergen-specific IgE against Ani s 1, and the diagnostic value of testing for antibodies to Ani s 1 has been extensively demonstrated. However, no previous studies have investigated the molecular aspects of the allergic response to Ani s 1. Knowledge of allergen-specific T cell and B cell (IgE and IgG4) epitopes is important for elucidating the immunological mechanisms underlying allergic responses, and for understanding why particular proteins behave as allergens. OBJECTIVE: To elucidate the main T cell- and B cell (IgE and IgG4)- binding regions of Ani s 1. METHODS: T cell epitopes were identified by peptide proliferation assays using T cell lines derived from peripheral blood mononuclear cells of 11 patients with Anisakis allergy, and IgE and IgG4 epitopes were identified by microarray immunoassay using sera from a different group of 11 patients with Anisakis allergy. RESULTS: Several T cell epitopes of Ani s 1 were identified, of which Ani s 1145-156 , Ani s 1151-162 and Ani s 1163-171 located at the C-terminal end of the protein were the most relevant. IgE and IgG4 recognized largely the same peptides, including Ani s 122-41 , Ani s 125-44 , Ani s 127-47 , Ani s 137-56 and Ani s 194-113 . CONCLUSIONS AND CLINICAL RELEVANCE: This is the first report describing the T cell epitopes of an important allergen of A. simplex, and the first B cell epitope study of this allergen in the Spanish population. This information can help to elucidate the mechanisms underlying the allergic response to Ani s 1, potentially leading to therapeutic and diagnostic advances.

Effects of a low-fat diet with antioxidant supplementation on biochemical markers of multiple sclerosis long-term care residents.

INTRODUCTION: Multiple sclerosis (MS) treatment options are primarily limited to immunomodulatory therapies in MS non-progressive forms. Nutrition intervention studies suggest that diet may be considered as a complementary treatment to control disease progression. Therefore, dietary intervention may help to improve wellness and ameliorate symptoms of MS patients. OBJECTIVES: To assess the effect of a low-fat diet with antioxidant supplementation on biochemical markers of institutionalized patients with progressive forms of multiple sclerosis. METHODS: A randomized prospective placebo-controlled study involving 9 participants, 5 of them assigned to the intervention group (low-fat diet and antioxidant supplementation) and the other 4 to the placebo group (low-fat diet). The effect of the dietary intervention, involving diet modification and antioxidant supplementation, was examined for 42 days by measuring anthropometric, biochemical parameters and oxidative stress markers in blood at baseline (day 0), intermediate (day 15) and end (day 42) stages of the treatment. RESULTS: The intervention group obtained C reactive protein levels significantly lower than those observed in the corresponding placebo group at the end of the study. Oxidative stress and inflammatory markers isoprostane 8-iso-PGF2α and interleukine IL-6 values also diminished after dietary intervention in the intervention group. Catalase activity increased significantly in the intervention group prior antioxidant supplementation. No significant differences were observed in other oxidative stress markers. CONCLUSIONS: The results suggest that diet and dietary supplements are involved in cell metabolism modulation and MS-related inflammatory processes. Consequently, low fat diets and antioxidant supplements may be used as complementary therapies for treatment of multiple sclerosis.

Introducción: Las posibilidades de tratamiento de la esclerosis múltiple (EM) se encuentran limitadas principalmente a terapias con inmumoduladores en las formas no progresivas de EM. Los estudios de intervención nutricional sugieren que la dieta puede considerarse como un tratamiento alternativo para controlar la progresión de la enfermedad. Por esta razón, las intervenciones en la dieta pueden ayudar a mejorar el bienestar y mejorar los síntomas de los pacientes con EM. Objetivos: Valorar el efecto de una dieta pobre en grasas con suplementación de antioxidantes en los marcadores bioquímicos de pacientes institucionalizados que presentan formas progresivas de EM. Métodos: Se realizó un estudio prospectivo aleatorizado controlado por placebo con 9 participantes, 5 de los cuales se asignan al grupo de intervención (dieta baja en grasas y suplementación antioxidante) y los 4 restantes al grupo placebo (dieta baja en grasas). Se evaluó el efecto de la intervención dietética que supone modificación de la dieta e introducción de antioxidantes durante 42 días mediante valoraciones de parámetros antropométricos y bioquímicos y marcadores del estrés oxidativo en sangre y orina en las etapas inicial (día 0), intermedia (día 15) y final (día 42) del tratamiento. Resultados: Se obtuvieron niveles de proteína C reactiva significativamente inferiores en el grupo de intervención con respecto al grupo placebo al final del estudio. Los marcadores de estrés oxidativo e inflamación: isoprostanos 8-iso-PGF2e interleucina IL-6 también disminuyeron en el grupo de intervención después de la intervención dietética. La actividad de la enzima catalasa aumentó de forma significativa en el grupo de intervención antes de la suplementación con antioxidantes. No se observaron diferencias significativas en otros marcadores de estrés oxidativo. Conclusiones: Los resultados obtenidos sugieren que la dieta y los suplementos dietéticos están involucrados en la modulación del metabolismo celular y los procesos de inflamación de la EM. En consecuencia, las dietas bajas en grasas y los suplementos antioxidantes podrían ser utilizados como terapias alternativas en el tratamiento de la EM.

Contact angioedema and rhinoconjunctivitis caused by Dendrobaena species and Sarcophaga carnaria used as fishing bait.

The flesh fly Sarcophaga carnaria is commonly used as fishing bait. Immunoglobulin (Ig) E-mediated reactions caused by the handling of this bait have been reported. The earthworm Dendrobaena species is increasingly being used as fishing bait but there have been no reported cases of allergy to this species to date. We studied a 26-year-old amateur angler who presented rhinoconjunctivitis, urticaria, and angioedema on handling S carnaria. He started to use Dendrobaena species instead but developed the same symptoms. The aim of this study was to identify the allergens involved in the patient's clinical reactions. The study was performed using immunoglobulin (Ig) E immunoblotting and immunoblotting inhibition assays.The patient's serum detected allergens from Dendrobaena species (of an apparent molecular weight of approximately 150, 60, 37, 24, 21 and 19 kDa) and S. carnaria (approximately 70 kDa and a smear ranging from 50 to 40 kDa). The patient was diagnosed with allergy to both Dendrobaena species and 5 carnaria. This is the first case describing Dendrobaena species as an allergic agent.

Effect of orlistat on lipid peroxidation, Na+, K+ ATPase, glutathione and serotonin in rat brain.

The aim of this study was to evaluate the effect of orlistat, a drug used in weight loss, on 5-HT and indicators of oxidative stress in rat brain. Orlistat, 12 mg/kg was administered to Wistar rats as single dose or successive doses on 3 consecutive days. Blood glucose and oxidative stress indicators were detected by measurement of lipid peroxidation, Na+, K+ ATPase, glutathione and serotonin levels using previous validated methods. The levels of glucose decreased in rats receiving successive doses. The activity of Na+, K+ ATPase and total ATPase was reduced in rats receiving successive doses, while the level of lipid peroxidation increased slightly in both groups. Glutathione underwent significant reduction in the successive doses group (p < 0.05). 5-HT increased significantly after single dose treatment (p < 0.05). Orlistat can induce pro-oxidant effects in the brain due to alteration of serotonergic metabolism and the reduction of glutathione.

Selective allergy to lobster in a case of primary sensitization to house dust mites.

Allergy to only 1 kind of seafood is uncommon. We report a case of selective allergy to lobster. We studied a 30-year-old man who suffered generalized urticaria, facial erythema, and pharyngeal pruritus after eating lobster. He had a more than 10-year history of mild persistent asthma and sensitization to house dust mites. The study was performed by skin prick test, and prick-prick test, oral food challenge, specific immunoglobulin (Ig) E determinations by CAP (Phadia, Uppsala, Sweden) and ADVIA-Centaur (ALK-Abelló, Madrid, Spain), and IgE-immunoblotting. The patient's serum recognized 2 allergens of around 198 kDa and 2 allergens of around 65 kDa from the lobster extract, allergens of around 15, 90, and 120 kDa from Dermatophagoides pteronyssinus extract, and allergens of around 15 and 65 kDa from Dermatophagoides farinae extract. Serum did not recognize purified shrimp tropomyosin. Immunoblot-inhibition assay results indicated cross-reactivity between lobster and mite allergens. This is the first report of selective allergy to lobster.

Lettuce-induced anaphylaxis. Identification of the allergen involved.

BACKGROUND: Only 2 allergenic proteins have been described in lettuce allergy: a 16-kDa protein (putative profilin) and a lipid transfer protein (LTP) named Lac s 1. OBJECTIVE: Our aim was to identify the allergens involved in the anaphylactic reactions of 2 patients who had eaten lettuce. METHODS: The study was performed by Ig (immunoglobulin)-E immunodetection and immunodetection-inhibition assays. RESULTS: Both patients' sera showed specific IgE binding to a single protein from the crude lettuce extract (apparent molecular weight of 14 kDa). To characterize the allergen detected, the lettuce extract underwent proteolytic digestion and heat treatment and was highly resistant to both. The patients' sera also recognized the major peach allergen Pru p 3 by immunodetection. When the lettuce allergen was incubated with both Pru p 3 from peach peel and recombinant Pru p 3, the immunodetection-inhibition assay indicated that patients were sensitized to the lettuce LTP Lac s 1. CONCLUSIONS: The allergen involved in the lettuce-induced anaphylaxis of our patients was the LTP Lac s 1.

Sensitization to serum albumins in children allergic to cow's milk and epithelia.

Patients with persistent milk allergy and specific immunoglobulin E (IgE) to bovine serum albumin (BSA) have a greater risk of rhinoconjunctivitis and asthma because of animal dander. To prove the cross-reactivity between serum albumin (SA) of different mammals in milk, meat, and epithelia and determine if heat treatment of meats decrease the allergenicity of albumins. The study was performed using SDS-PAGE and IgE-immunoblotting using sera from eight patients sensitized to milk, BSA, and animal danders. Sera from non-allergic and only animal dander allergic subjects served as a control. With one exception, all patients' sera recognized SA in different meats (beef, lamb, deer, and pork), epithelia (dog, cat, and cow), and cow's milk. Some patients even were only sensitized to SA in meat and epithelia. Danders' allergic only recognized other proteins in epithelia but not SA. No patients reacted to SA from heated meat extracts. Serum albumin is an important allergen involved in milk, meat, and epithelia allergy. The first contact with SA was through cow's milk and patients developed sensitization to epithelia SA even without direct contact with animals. Patients with both BSA and cow's milk allergy must avoid raw meats and furry pets.

Modelling the growth of Leuconostoc mesenteroides by Artificial Neural Networks.

The combined effect of temperature (10.5 to 24.5 degrees C), pH level (5.5 to 7.5), sodium chloride level (0.25% to 6.25%) and sodium nitrite level (0 to 200 ppm) on the predicted specific growth rate (Gr), lag-time (Lag) and maximum population density (yEnd) of Leuconostoc mesenteroides under aerobic and anaerobic conditions, was studied using an Artificial Neural Network-based model (ANN) in comparison with Response Surface Methodology (RS). For both aerobic and anaerobic conditions, two types of ANN model were elaborated, unidimensional for each of the growth parameters, and multidimensional in which the three parameters Gr, Lag, and yEnd are combined. Although in general no significant statistical differences were observed between both types of model, we opted for the unidimensional model, because it obtained the lowest mean value for the standard error of prediction for generalisation. The ANN models developed provided reliable estimates for the three kinetic parameters studied; the SEP values in aerobic conditions ranged from between 2.82% for Gr, 6.05% for Lag and 10% for yEnd, a higher degree accuracy than those of the RS model (Gr: 9.54%; Lag: 8.89%; yEnd: 10.27%). Similar results were observed for anaerobic conditions. During external validation, a higher degree of accuracy (Af) and bias (Bf) were observed for the ANN model compared with the RS model. ANN predictive growth models are a valuable tool, enabling swift determination of L. mesenteroides growth parameters.

Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2.

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach. METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity. RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01. CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.