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Kerry, Ireland. Hazelnut flour (30% protein) was purchased from Nutt Ranch, Marlborough, New Zealand; and celeriac powder (11.8% protein) from Kraeuter Mix GmbH, Abstwind, Germany. All other chemicals were of analytical grade, and distilled water was used unless otherwise stated. A chocolate dessert base formulation was developed by the Institute of Food Research, based on a system devised by Unilever R&D BV (Vlaardingen, Netherlands). For each allergen, a ��high�� and ��low�� allergen dessert base was produced to enable delivery of the required dose range. In brief, rapeseed oil and Tween 60 were mixed, warmed S6 Kinase to approximately 40��C until the mixture became homogenous, and the resultant was added to dry ingredients and mixed by large domestic mixer (Kenwood Chef; Kenwood Ltd, Havant, UK) (2?min minimum speed, 5?min maximum speed). The resulting paste was dispensed in 30 g aliquots into sterile 125?ml containers (VWR International, West Sussex, UK) and packed into kits comprising ��high-allergen�� (three pots), ��low-allergen�� (one pot) and placebo (allergen-free; five pots) dessert bases, dosing spoons and instructions for storage and use. Prior to use, dessert base was hydrated with still, bottled mineral water (70?ml per 30?g of dessert base), mixed until it became smooth and chilled at 2�C5��C, for at least 30?min, to allow the cold swelling starch to fully hydrate. The resulting ��mousse�Cstyle�� dessert was mixed again to ensure homogeneity and stored Selleckchem AZD-3965 at 2�C5��C for a maximum of 24?h prior to DBPCFC. The recipes for hydrated (as-eaten) placebo, high-allergen and low-allergen desserts are described in Table?1a (hazelnut), b (celeriac). Desserts are classed as a category 1 food, according to UK guidelines for microbiological quality (23), and had to pass microbiological testing prior to use. Dessert bases were analysed in triplicate at Unilever (Colworth, Bedford, UK) for total colony counts (acceptable limits: check details