Peanut oleosins associated with severe peanut allergy-Importance of lipophilic allergens for comprehensive allergy diagnostics.
BACKGROUND: Peanut allergy is one of the most common and most severe food allergies in Western countries and its accurate diagnosis to prevent potential life-threatening allergic reactions is crucial. However, aqueous extracts used for routine diagnostic measurements are devoid of lipophilic allergens such as oleosins. We have recently succeeded in the isolation and purification of these unique proteins, and the current study evaluates their allergenic potential and clinical relevance. OBJECTIVE: We sought to assess allergenicity and sensitization prevalence of oleosins obtained from both raw and in-shell roasted peanuts. Additionally, we tested the utilization of natural and recombinant oleosins for allergy diagnostic purposes. METHODS: Oleosin sensitization, prevalence, and impact of thermal processing were analyzed by immunoblot with sera from 52 peanut-allergic individuals displaying different clinical phenotypes. The application of natural and recombinant oleosins for allergy diagnostics was investigated by basophil activation test (BAT). IgE-binding epitopes were identified by oligopeptide microarray. RESULTS: Sensitization to oleosins was observed exclusively in peanut-allergic subjects suffering from severe systemic reactions. IgE-binding capacity of oleosins derived from in-shell roasted peanuts was increased as shown by immunoblot analysis and BAT. Both natural and recombinant molecules can be used to identify oleosin-sensitized patients by BAT. A linear epitope of Ara h 15 was determined which displays high similarity to other seed-derived oleosins. CONCLUSION: Oleosins are clinically relevant peanut allergens and most likely associated with severe allergic symptoms. In-shell roasting increases their allergenicity, which is consistent with the observation that most allergic reactions are in connection with roasted peanuts.
Effects of sulfur dioxide concentration on organic acids and β-carotene in dried apricots during storage.
The effects of various sulfur dioxide (SO2) concentrations (0, 451, 832, 1594, 2112 and 3241mg/kg) on the profiles and contents of organic acids (OAs) and β-carotene in sulfured dried apricots (SDAs) were investigated during storage at 4, 20 and 30°C for 379days. In all samples, four OAs [malic acid (MA), citric acid (CA), succinic acid (SA) and oxalic acid (OXA)] were identified. SA (13.9-31.8g/kgdw) was the major OA in SDAs containing SO2 at lower than 1594mgSO2/kg, while MA (11.7-11.7g/kgdw) was the major OA in SDAs containing SO2 at higher than 1594mgSO2/kg. As SO2 concentration increased, CA and OXA contents increased whereas MA contents decreased. Moreover, the highest stabilities of β-carotene, MA and SO2 were determined in SDAs containing 1594mgSO2/kg at 4°C. Therefore, we suggest using 1594mgSO2/kg and storing SDAs at 4°C to protect OAs and β-carotene.
Update on Timing and Source of ‘Allergenic' Foods.
As the prevalence of food allergies in many communities continues to rise, the question of when in infancy to introduce any solid foods, or specific 'more allergenic' foods, as a food allergy prevention strategy has been debated. Observational studies have found that introducing any solid foods prior to 4 months of age was associated with an increased risk of allergic disease. Hence, the current allergy prevention consensus recommendation is that the introduction of any solid foods should commence after 4 months of age. Over recent years, several randomized controlled trials have been conducted to investigate the 'ideal' timing of introduction of some specific 'more allergenic' foods (including peanuts and eggs) into infant diets. To date, the results from three of these trials have determined that there is no reason to delay the introduction of the 'more allergenic' foods into the infant's diet after solid foods have commenced. However, these trials have also highlighted the finding that some infants are sensitized to food allergens before any known ingestion of solid foods. Thus, future research needs to focus on strategies to prevent early-life food allergen sensitization prior to complementary feeding.
Conjunctival provocation test in diagnosis of peanut allergy in children.
BACKGROUND: Peanut allergy frequently cause severe allergic reactions. Diagnosis include detection of IgE to peanuts in serum or by skin prick tests. While children may have allergic sensitisation without having clinical peanut allergy, oral peanut challenge is often required for accurate diagnosis. The conjunctival provocation test is used for diagnosis and evaluation of treatment effect in inhalant allergies, but it has not been evaluated as a tool for diagnosing peanut allergy. OBJECTIVE: To investigate if the conjunctival provocation tests may be feasible, accurate and safe in diagnosing clinically relevant peanut allergy in patients with suspected peanut allergy. METHODS: This cross-sectional case-control study in children with clinical or laboratory suspected peanut allergy included 102 children recruited from the regional paediatric departments and specialist practices during one year from April 2011. A peanut tolerant control group of 28 children of similar age was recruited locally. A double-blind placebo-controlled conjunctival provocation test with peanut extract was performed in all children, while oral peanut provocation was performed as double-blind placebo-controlled challenge in children with suspected peanut allergy and as an open challenge in the control children. RESULTS: All 81 children with a positive double-blind placebo-controlled oral food challenge also had a positive conjunctival provocation test. None of the children with negative conjunctival provocation test had a positive oral food challenge. The sensitivity and the specificity of the conjunctival provocation test was 0.96 and 0.83 respectively. No children had severe adverse reaction caused by the conjunctival provocation test, whereas 23 children suffered an anaphylactic reaction to the oral food challenge. CONCLUSION & CLINICAL RELEVANCE: Conjunctival allergen challenge appears to be a feasible, accurate and safe in diagnosing children referred for suspected peanut allergy.
Low percentage of clinically relevant pistachio nut and mango co-sensitisation in cashew nut sensitised children.
BACKGROUND: Cashew nut, pistachio nut and mango belong to the Anacardiaceae family and are botanically related. Therefore, cashew nut sensitised children are frequently advised to eliminate cashew nuts and pistachio nuts from their diet. The 'Improvement of Diagnostic mEthods for ALlergy assessment (IDEAL trial number NTR3572) study showed that cashew nut sensitised children were co-sensitised to pistachio nut in 98% of cases and to mango in 21% of cases. The aim of this follow-up study to IDEAL is to assess the clinical relevance of co-sensitisation to pistachio nut and mango in cashew nut sensitised children. METHODS: Children were recruited from the study: 'Improvement of Diagnostic mEthods for ALlergy assessment (IDEAL trial number NTR3572). Inclusion criterion for the IDEAL study was sensitization to cashew nut as demonstrated by either SPT or sIgE, and a clinical history of reactions to cashew nuts or no previous (known) exposure. Sensitized children who were tolerant to cashew nuts were excluded. Inclusion criterion for this IDEAL follow-up study was co-sensitization to pistachio nut, regardless the result of the DBPCFC with cashew nut. In this follow-up study a double-blind placebo-controlled food challenge with pistachio nut and an open food challenge with mango were performed. RESULTS: Twenty-nine children (mean age of 11.6 years, 62% male) were included. Pistachio nut sensitisation was clinically relevant in only 34% of cashew-sensitised children and only 31% of cashew challenge positive children. None of the children was challenge positive to mango. CONCLUSION: Although co-sensitisation between cashew nut and pistachio nut was observed in 98%, pistachio nut sensitisation was only clinically relevant in 34% of the children. Therefore, a challenge test with pistachio nut is recommended in children with cashew nut and pistachio nut sensitisation. Trial registration The study was registered in the Dutch trial register (registration number 3572) on 10 August 2012 (retrospectively registered).
Early introduction of peanuts.
Peanut allergy is the most common food allergy and the leading cause of anaphylaxis and death due to food allergy. Despite previous guidelines created by the American Academy of Pediatrics in 2003, peanut allergy has continued to significantly increase over the past decade. Therefore, further research has been done to help clinicians provide more evidence-based recommendations about the timing of introduction of peanuts. The LEAP study, published in February 2015, demonstrated the value of much earlier introduction of peanuts to nonallergic patients than previously suggested. These findings have altered current practice, and recommendations supported by the American Academy of Pediatrics now allow the introduction of peanuts as early as age 4 months to reduce the likelihood of developing peanut allergy.
Detection of Food Allergens by Taqman Real-Time PCR Methodology.
Real-time PCR (polymerase chain reaction) has shown to be a very effective technology for the detection of food allergens. The protocol described herein consists on a real-time PCR assay targeting the plant ITS (Internal Transcribed Spacer) region, using species-specific primers and hydrolysis probes (Taqman) dual labeled with a reporter fluorophore at the 5' end (6-carboxyfluorescein, FAM) and a quencher fluorophore at the 3' end (Blackberry, BBQ). The species-specific real-time PCR systems (primers/probe) described in this work allowed the detection of different nuts (peanut, hazelnut, pistachio, almond, cashew, macadamia, walnut and pecan), common allergens present in commercial food products, with a detection limit of 0.1 mg/kg.
Validation of Recipes for Double-Blind Placebo-Controlled Challenges With Milk, Egg White, and Hazelnut.
BACKGROUND: The double-blind, placebo-controlled food challenge (DBPCFC) is considered the definitive diagnostic test for food allergy. Nevertheless, validated recipes for masking the foods are scarce, have not been standardized, and differ between centers. Sensory evaluation techniques such as the triangle test are necessary to validate the recipes used for DBPCFC. METHODS: We developed 3 recipes for use in DBPCFC with milk, egg white, and hazelnut and used the triangle test to validate them in a 2-phase study in which 197 volunteers participated. In each phase, participants tried 3 samples (2 active-1 placebo or 2 placebo-1 active) and had to identify the odd one. In phase 1, the 3 samples were given simultaneously, whereas in phase 2, the 3 samples of foods that failed validation in phase 1 were given sequentially. A visual analog scale (VAS) ranging from 1 to 10 was used to evaluate how much participants liked the recipes. RESULTS: In phase 1, the egg white recipe was validated (n=89 volunteers, 38.9% found the odd sample, P=.16). Milk and hazelnut recipes were validated in phase 2 (for both foods, n=30 participants, 36.7% found the odd sample, P=.36). Median VAS scores for the 3 recipes ranged from 6.6 to 9.7. CONCLUSIONS: We used sensory testing to validate milk, egg white, and hazelnut recipes for use in DBPCFC. The validated recipes are easy to prepare in a clinical setting, provide the equivalent of 1 serving dose, and were liked by most participants.
Detection of Food Allergens by Phage-Displayed Produced Antibodies.
Phage display is a powerful tool to produce recombinant antibodies against a given antigen without animal immunization. This technology employs libraries of recombinant bacteriophages that display billions of different functional antibody fragments on their surface. They are selected by panning in vitro against the target antigen in search for specific binders. In this chapter, we describe the selection of single chain variable fragment (scFv) antibodies to be used for detection of allergenic proteins from nuts in food products. The artificial libraries TomLinson I+J (MRC Laboratory of Molecular Biology and MRC Centre for Protein Engineering) were employed that resulted in successful phage-ELISA systems for detection of almond and walnut proteins in commercial food products.
Drying and decontamination of raw pistachios with sequential infrared drying, tempering and hot air drying.
Pistachio nuts have been associated with outbreaks of foodborne disease and the industry has been impacted by numerous product recalls due to contamination with Salmonella enterica. The current hot air drying of pistachios has low energy efficiency and drying rates, and also does not guarantee the microbial safety of products. In the study described herein, dehulled and water-sorted pistachios with a moisture content (MC) of 38.14% (wet basis) were dried in a sequential infrared and hot air (SIRHA) drier to <9% MC. The decontamination efficacy was assessed by inoculating pistachios with Enterococcus faecium, a surrogate of Salmonella enterica used for quality control in the almond industry. Drying with IR alone saved 105min (34.4%) of drying time compared with hot air drying. SIRHA drying of pistachios for 2h with infrared (IR) heat followed by tempering at a product temperature of 70°C for 2h and then by hot air drying shortened the drying time by 40min (9.1%) compared with drying by hot air only. This SIRHA method also reduced the E. faecium cell population by 6.1-logCFU/g kernel and 5.41-logCFU/g shell of pistachios. The free fatty acid contents of SIRHA dried pistachios were on par with that of hot air dried samples. Despite significant differences in peroxide values (PV) of pistachio kernels dried with the SIRHA method compared with hot air drying at 70°C, the PV were within the permissible limit of 5Meq/kg for edible oils. Our findings demonstrate the efficacy of SIRHA drying in achieving simultaneous drying and decontamination of pistachios.