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A 30-year-old man from the Philippines with pollen allergy noted the appearance of oral allergy syndrome (OAS) after eating raw apple, raw peach, raw celery, and recently, jackfruit (Artocarpus integrifolia), a tropical fruit which belongs to the Moraceae family (mulberry) and to the genus Artocarpus (breadfruit tree). Despite the patient's multiple sensitization in skin prick tests and in the Pharmacia CAP System to birch, grass, mugwort pollen, related fruits and vegetables, and jackfruit, in RAST-inhibition studies neither rBet v 1 nor rBet v 2 (profilin), the well-known cross-reacting allergenic components in OAS, could inhibit the specific IgE response to jackfruit. Whether the reaction to jackfruit is specific or whether other pollen-related, cross-reacting allergenic components exist should be investigated further.

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BACKGROUND: Oral allergy syndrome in response to fruits and vegetables frequently occurs as clusters of hypersensitivity to members of the same botanical family, for which the immunologic basis lies in a number of common allergens, most of them still unidentified. OBJECTIVE: This study was designed to assess the in vivo and in vitro cross-reactivity between fruits of the Prunoideae subfamily (i.e., peach, cherry, apricot, and plum) and to identify their major allergens and the cross-reactivity of the peach extract with grass and birch pollen. METHODS: The in vivo study was conducted by skin prick tests and open food challenges with fresh fruits in 23 patients with oral allergy syndrome for peach and positive skin prick test and RAST results for the other Prunoideae. In vitro sodium dodecylsulfate-polyacrylamide gel electrophoresis was followed by immunoblotting and immunoblotting-inhibition. RESULTS: A 13 kd component was identified as the only major allergen common to all the Prunoideae, the other major allergens were found at 14 kd in peach and at 30 kd in cherry. Immunoblotting inhibition showed wide cross-reactivity within the Prunoideae, whereas grass and birch pollen partially inhibited the peach blotting. CONCLUSIONS: Clinical cross-reactivity to Prunoideae is essentially due to a common 13 kd IgE-binding component, which seems to be the most important major allergen of this subfamily, not shared with grass and birch pollen.

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Aspergillus fumigatus grown in submerged and surface cultures was extracted, and the extracts were analyzed separately. The submerged extract contained 31.9% protein and 8.3% carbohydrate, while the corresponding values were 17.0% and 33.3% for the surface material. With individual sera from patients with allergic asthma, SDS-PAGE combined with immunoblotting revealed that the submerged extract contained at least six strong IgE-binding components (20, 30, 38, 50, 68, and 90 kDa) in addition to several weak to medium IgE-binding components. The surface extract contained about the same number of IgE-binding components, but only one gave a strong reaction (20 kDa). The allergens present were shown to have pI between 4.5 and 5.6 as demonstrated by isoelectric focusing (IEF) combined with immunoblotting. For identification of A. fumigatus glycoprotein allergens, both extracts were treated with periodate under mild conditions. Two allergens of the submerged extract (90 and 38 kDa) partly lost their IgE-binding ability by this treatment, indicating that these components are glycoproteins and that the carbohydrate moiety is involved in the IgE binding. The IgE-binding ability of the 20-kDa allergen was not influenced by periodate. For assessment of the stability of the two allergen extracts, aqueous solutions were kept at 4 degrees C for 2, 7, and 21 d and then analyzed by SDS-PAGE and immunoblotting. The results showed that most allergens of the submerged extract were partly inactivated after 2 d. After 21 d, only the 20-kDa and 30-kDa components were still able to bind IgE. Similar results were obtained by analyzing the surface extract.(ABSTRACT TRUNCATED AT 250 WORDS)

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Allergy to kiwi, poppy seeds, and/or sesame seeds often occurs in patients with a simultaneous sensitization to nuts and flour. Previously cross reactions have been verified by RAST inhibition. In this study the nature of this cross-reactivity is further characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), followed by immunoblotting to nitrocellulose. The degree of cross-reactivity among kiwi, sesame seeds, poppy seeds, hazelnuts, and rye grain was found to be very high in the patients studied. The existence of both cross-reacting and unique components was observed; however, the cross-reacting and unique components could be different for different patients.

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The allergenic similarity of the pyroglyphid mite D. pteronyssinus and the glycyphagid mite L. destructor was investigated with a new immunoblotting inhibition technique allowing simultaneous comparison of several allergens. Extracts of D. pteronyssinus and L. destructor were separated by SDS-PAGE and electroblotted to nitrocellulose (NC). A serum pool containing IgE specific to the major allergens in both mites was mixed with serially diluted extracts of D. pteronyssinus and L. destructor and incubated with the mite allergens of NC. The inhibition of the IgE binding to NC was evaluated by densitometric scanning and percentage inhibition was calculated. The IgE antibodies to the 25-kD component in D. pteronyssinus, were inhibited to the same degree by extracts of D. pteronyssinus and L. destructor. Another major allergen component in D. pteronyssinus (16 kD) was also inhibited by L. destructor extract but to a lesser degree: 400 times more of the heterologous than of the homologous extract was needed for 50% inhibition. To produce 50% of heterologous inhibition of the two major allergen components at 15 and 53 kD of L. destructor, 2000 and 10,000 times more respectively, of D. pteronyssinus than of L. destructor extract were needed. Two minor allergen components of L. destructor showed some cross-reactivity with D. pteronyssinus. However, L. destructor was a stronger inhibitor of D. pteronyssinus than vice versa, probably because the sera were obtained from persons more sensitized to L. destructor than to D. pteronyssinus.

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Sera from patients sensitized to Aspergillus fumigatus (A.f.) were screened for specific IgE using sodiumdodecylsulphate-gradient-polyacrylamidegelelectrophoresis (SDSgPAGE) followed by immunoblotting to nitrocellulose. Approximately 25 IgE-binding components were detected. The components of molecular weight 20, 31, 44, 50, 53, 77 and 90 kD were reacting with more than 50% of the patients. The 90, 77 and 20-kD components showed up as the strongest IgE-binding bands. The 20-kD component, called Ag 20 kD, was purified and further characterized. Ag 20 kD was purified to apparent homogeneity. using a combination of size-exclusion chromatography on a Sephacryl S-200 column, preparative isoelectric focusing in a pI 2.5-6.5 gradient, and a Sephadex G-50 Superfine column. Fractions were characterized with protein and carbohydrate analyses, RAST and SDSgPAGE followed by immunoblotting to nitrocellulose. Ag 20 kD was found to be a glycoprotein as it stained with both Coomassie Brilliant Blue and PAS. However, it did not bind Con A, and thus, did probably not contain any terminal alpha-D mannopyranosyl end groups. The relation between mannose, galactose and glucose was found to be 2:1:0.5. The isoelectric point was heterogeneous within pH range 5-6, and the molecular weight was estimated to approximately 20 kD. An increased RAST response was shown for the purified component compared with crude extract using patient sera reacting with Ag 20. The antigen was shown not to be identical with the previously described Ag 3. Neither did it fit the description of Ag 5, 7 or 13 earlier described by the same group. The antigen is going to be used for further immunochemical and clinical investigations, and coupling to other systems for antigen characterization.

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The effects of sample treatments, separation conditions, and the possible presence of antibody interference phenomena in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE) followed by nitrocellulose immunoblotting (IB) were studied using allergens from Alternaria alternata (Alt.a.) and Aspergillus fumigatus (Asp.f.) as model systems. In order to obtain good resolution in the IB method the mould allergens were separated in gradient gels under dissociating conditions (SDSgPAGE) including sample treatment with boiling, SDS and 2-mercaptoethanol (ME). These treatments all reduced the IgE binding capacities of the Asp.f. and Alt.a. extracts studied. Nevertheless, a great variety of IgE-binding components were detected after IB, and this could probably be explained by refolding of allergens during the blotting procedure where SDS was partly removed. A comparison of IgG-enriched and IgG-reduced serum fractions in IB revealed small differences in the IgE-binding patterns suggesting that IgG interference is of minor importance in the system studied. The IB method must be individually optimized for each new allergen studied using well characterized sera, preferably monospecific for the major components. Taking these facts into consideration, the IB technique is a valuable complement to other methods in the study of mould allergens.

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A panel of 55 sera from patients with suspected mold allergy from two different geographic areas, A and B, was investigated for specific IgE antibodies to 16 individual molds using Phadebas RAST technique. The molds investigated were Alternaria, Cladosporium, Aspergillus, Penicillium, Mucor, and Candida from the former RAST panel and ten additional new mold genera. The RAST screening revealed that 73% of patients had specific IgE antibodies to at least one of the 16 molds. Eighteen percent were negative to the six molds of the previous RAST panel but had specific IgE antibodies to at least one of the ten new molds. The highest frequency of IgE antibodies in the A group could be assigned to Cladosporium, followed by Botrytis and Helminthosporium. In group B, sensitivity to Botrytis and Phoma showed the highest frequencies. This study revealed that the relative importance of mold genera may vary greatly between patient groups from different geographic areas. Some of the species of the expanded RAST panel seem to be at least as important as Alternaria and Cladosporium. Multiple sensitivities seem to be caused by sensitization by many mold species rather than by cross-reactivity.

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A random sample of 46 general practitioners of the Unitá Sanitaria Locale in Torino recruited 802 elderly outpatients and collected information about complaints and current drug treatment. Within a week each patient received a home interview and details were collected on drug compliance and use of drugs other than those reported by the general practitioners. On average, each patient was taking 3.6 drugs, of which 2.9 were correctly reported by the general practitioners and 0.7 were unreported. Among the most prescribed therapeutic groups there were drugs with a narrow therapeutic index (cardiovascular drugs, diuretics, psychotropic agents) and substances whose efficacy has never been fully documented ("cerebroactive-vasoactive" agents). Age and number of complaints were positively and significantly correlated with number of prescribed drugs. Nearly half of the sample (44.4%)--more frequently women and people with higher education--were taking one or more drugs not detected by the general practitioners, often benzodiazepines taken over a long period for anxiety or insomnia. Full compliance occurred for 81.5% of the prescriptions and 59.9% of patients were correctly taking all prescribed drugs. Compliance was lower for prescriptions of the general practitioners compared with other doctors' prescriptions (eg, hospital doctor, private doctor) and probability of taking correctly all the prescribed drugs decreased with the number of medicines concurrently taken. The most common reason for noncompliance was fear of side effects.

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A random sample of 46 general practitioners of the Unità Sanitaria Locale in Torino recruited 802 elderly outpatients and collected information about complaints and current drug treatment. Within a week each patient received a home interview and details were collected on drug compliance and use of drugs other than those reported by the GP. On average, each patient was taking 3.6 drugs of which 2.9 were correctly reported by the GP and 0.7 were un reported. Among the most prescribed therapeutic groups there were drugs with a narrow therapeutic index (cardiovascular drugs, diuretics, psychotropic agents) and substances whose efficacy has never been fully documented ("cerebroactive/vasoactive"). Age and number of compliants were positively and significantly correlated with number of prescribed drugs. The most common reason for non-compliance was fear of side-effects.

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