Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens.

BACKGROUND: Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia-sensitized patients or its relationship with other LTP allergens is not clear. OBJECTIVE: To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area. METHODS: Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens. RESULTS: The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3. CONCLUSION: Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent.

Pressurised metered-dose inhalers (MDIs) versus dry powder inhalers devices (DPIs) to rapid-acting inhaled b2-agonists for asthma in children.

OBJECTIVE: To compare the clinical effectiveness of pressurized metered-dose inhalers (MDIs) with that of dry powder inhalers (DPIs) in delivering short-acting b2-agonists in children with asthma. METHODS: Searches were performed in Medline (1997-March 2002), the Cochrane Library Database and the Embase reference lists of review articles and clinical trials. In addition, the international headquarters of b2-agonist manufacturers were contacted. We performed a review of randomized controlled trials. RESULTS: Ten randomized controlled trials were included. No differences in clinical effectiveness were found between MDIs and PDIs. Two studies reported that fewer adverse events occurred when the Turbuhaler was used. Two long-term studies in children found that children preferred the MDI to the Rotohaler. CONCLUSIONS: 1) In stable asthma, short-acting b2 bronchodilators in standard MDIs are as effective as dry powder inhalers. 2) Pooling of results was limited by the small number of studies and therefore no overall conclusions could be drawn. 3) From the limited data available, we found little or no evidence for an additional clinical benefit of DPI devices over standard MDIs in children with asthma.

cDNA cloning and heterologous expression of the major allergens from peach and apple belonging to the lipid-transfer protein family.

BACKGROUND: Lipid-transfer proteins (LTPs) have been identified as major allergens of Rosaceae fruits in populations living in areas virtually free of Fagales trees, such as several Mediterranean communities. Pru p 3 and Mal d 3, the allergens from peach and apple, respectively, have a main clinical relevance in these areas. OBJECTIVE: To isolate and characterize cDNAs for Pru p 3 and Mal d 3, and to produce recombinant Pru p 3 in the yeast Pichiapastoris. METHODS: cDNAs for both allergens were isolated by polymerase chain reaction using nondegenerated primers. Expression of Pru p 3 was performed in P. pastoris using the pPIC 9 vector. The recombinant product was purified by gel-filtration chromatography followed by RP-HPLC. Immunodetection and immunoblot inhibition assays were carried out with sera from peach-allergic patients. RESULTS: The cDNAs for both Pru p 3 and Mal d 3 showed a 273 open reading frame coding for the 91 amino acid mature polypeptides. The deduced amino acid sequences exhibited N-terminal regions fully identical to those previously determined for the natural peach and apple allergens. Pru p 3 was expressed in P. pastoris at 20 mg/L of culture medium. The recombinant allergen showed the same N-terminal sequence (plus a glutamic acid added for proper extracellular expression) and apparent molecular size as natural Pru p 3. Both the recombinant and natural forms of Pru p 3 displayed similar immunoglobulin (Ig)E-binding capacity in immunodetection and immunoblot inhibition assays. CONCLUSIONS: Comparison of the complete primary structures of mature Pru p 3 and Mal d 3 deduced from their corresponding cDNA clones supports the close relationship between both allergens. Recombinant Pru p 3 binds IgE in vitro like its natural counterpart. Therefore, it can be a useful tool for specific diagnosis and structural studies.

Positividad de la prueba del parche con fenitoína en un caso de síndromede Stevens-Johnson / Positive patch test with phenytoin in a case of Stevens-Johnson syndrome

El síndrome de Stevens-Johnson (SJS) o eritema multiforme mayor es una reacción de posible mecanismo inmunológico poco frecuente, con una incidencia media anual estimada de 1-2 casos por millón de habitantes. La fenitoína y otros anticonvulsivantes, tales como fenobarbital y carbamnacepina, se han asociado con casos de SJS. Se describe el caso de una mujer de 40 arios de edad, diagnosticada de un tumor cerebral (oligodendroglloma), que presentó un cuadro clínico compatible con SJS, 48 horcas después de administrarle metamizol magnésico y fenitoína, por vía intravenosa (la paciente había realizado previamente un ciclo de tratamiento con buena tolerancia a dichos fármacos). Se realizaron pruebas cutáncas (1prick test) con los preparados comerciales de fenitoína (50 mg/ml) y metamizol (400 mg/ml), que frieron negativas en ambos casos. Se llevaron a cabo pruebas epicutáneas con los preparados comerciales de los fármacos implicados que resultaron positivas a fenitoína, a las 48 y 96 horas con persistencia de la lesión resultante incluso una semana después, y negativas en el caso del metamizol. La provocación oral hasta alcanzar la dosis terapéutica con metamizol fue tolerada. La provocación con fenitoína no se realizó, por el riesgo de una reacción potencialmente grave. Este caso puede servir como ejemplo de la utilidad de las pruebas epicutáneas para llegas- a identificar al agente causal, en casos de SJS relacionados con la toma de varios medicamentos (AU)

Lipid-transfer proteins as potential plant panallergens: cross-reactivity among proteins of Artemisia pollen, Castanea nut and Rosaceae fruits, with different IgE-binding capacities.

BACKGROUND: Lipid-transfer proteins (LTPs), but not Bet v 1 homologues, have been identified as major allergens of apple and peach in the Rosaceae fruit-allergic population in the Mediterranean area. Many of these patients show cosensitization to mugwort pollen. LTPs have an ubiquitous distribution in tissues of many plant species, and have been proposed as a novel type of plant panallergens. OBJECTIVE: We sought to isolate LTPs from Artemisia pollen and from a plant food not belonging to the Rosaceae family, such as chestnut nut, and to compare their amino acid sequences and IgE-binding capacities with those of apple and peach LTPs. METHODS: Allergens (LTPs) were isolated by different chromatographic methods (gel-filtration, ion exchange and/or reverse-phase HPLC), and characterized by N-terminal amino acid sequencing and MALDI analysis. Specific IgE-quantification and immunodetection, as well as immunoblot and ELISA inhibition assays, were carried out using sera from patients allergic to both apple and peach. RESULTS: Purified LTPs from Artemisia pollen and from chestnut seed showed molecular masses about 9 700d, and 43-50% sequence identity with the equivalent allergens of apple and peach in the first 30 N-terminal residues, which comprise about one third of the total amino acid sequence. A similar degree of sequence identity (50%) was found between the Artemisia and chestnut proteins. Both isolated LTPs bound specific IgE of sera from Rosaceae fruits allergic patients. However, substantially lower values of specific IgE-binding and maximum ELISA inhibition percentages were obtained for Artemisia and chestnut LTPs when compared to those from apple and peach. CONCLUSION: LTPs from Artemisia pollen and chestnut crossreact with allergens (LTPs) of Rosaceae fruits, but significant differences in specific IgE-binding capacities were observed among members of the plant LTP family. Thus, further studies are needed to evaluate the clinical significance of the observed cross-reactivities of plant LTPs.

Comparison of the efficacy and safety of two preseasonal regimens of glutaraldehyde modified, tyrosine-adsorbed parietaria pollen extract over a period of three years in monosensitive patients.

The purpose of this study was to evaluate the clinical efficacy over a period of three years (1988-90) of two preseasonal dosage regimens of a Parietaria allergoid (Bencard Tyrosine Parietaria) in patients who were only sensitive to this pollen. Fifty patients were included (14 men and 36 women, age: mean, 28 years; range, 14-47 years). Twenty five patients (group A) were treated each january with the basic course of Bencard Tyrosine Parietaria. This consisted of injecting subcutaneously 0.5 ml from each of three vials, with one week between each injection. A further injection using the vial with the highest dose was given one week later. Each january and february, twenty five patients (group B) were treated with the basic course of Bencard Tyrosine Parietaria, repeating the last dose five times, with one week between each injection. Immunotherapy with a tyrosine-adsorbed Parietaria judaica allergoid is an effective method for mitigating nasal (p < 0.0001), bronchial (p < 0.005), conjunctival (p < 0.001) and palatal itching symptoms (p < 0.0001) in patients who are sensitive to this pollen. Sensitivity to Parietaria pollen, as verified by skin test and nasal challenge, decreased during immunotherapy (p < 0.001). Histamine release by peripheral blood basophils decreased during the course of the study, falling from 43.5 ng/ml to 12.3 ng/ml in group A and from 42.9 ng/ml to 10.0 ng/ml in group B; during the second and third years, IgG levels were increased one and four months after starting treatment with the extract, while this was not the case after ten months; IgE levels were also increased. Finally, overall tolerance to this immunotherapy product was good in almost all patients.

Lipid-transfer proteins are relevant allergens in fruit allergy.

BACKGROUND: Allergy to apple and Prunus fruits is frequently associated with birch pollinosis, with the principal cross-reacting allergens involved being members of the Bet v 1 family. However, a major 13-kd component, nonimmunologically related to Bet v 1, has been implicated as allergen in patients allergic to Prunoideae fruit but not to birch pollen. OBJECTIVE: We sought to isolate and characterize the 13-kd allergen present in apple and peach. METHODS: Sera from patients allergic to both fruits were selected on the basis of clinical symptoms, skin prick tests responses, and specific IgE levels. Allergens were purified by reverse-phase HPLC and characterized by N-terminal amino acid sequencing, MALDI analysis, specific IgE immunodetection, and immunoblot inhibition assays. RESULTS: A 13-kd protein band was recognized in crude apple and peach extracts by 9 of 10 and 10 of 10 sera from patients allergic to fruit, respectively. The isolation and characterization of the corresponding allergens allowed their identification as lipid-transfer proteins, with a molecular mass of 9058 d for the apple protein and 9138 d for the peach protein. Both purified allergens were recognized by sera from patients allergic to fruit and fully inhibited the IgE binding by the 13-kd component present in the 2 crude fruit extracts. CONCLUSION: Lipid-transfer proteins are relevant apple and peach allergens and, considering their ubiquitous distribution in tissues of many plant species, could be a novel type of panallergen of fruits and vegetables.

The role of rhinovirus in allergic airway inflammation.

Epidemiological data demonstrate that viral infections are the most important trigger for acute asthma symptoms in children, and this association persists in many adults with asthma. Studies on volunteers experimentally infected with rhinoviruses (RV) suggest that atopy alone does not predispose to unusually severe symptoms. In contrast, experimental models combining viral infection and allergen exposure have identified potential links between virus-induced and allergen-induced inflammation. While in vitro studies suggest that cytokines may be an important part of this association, their role must be verified by sampling lower airway fluids and tissues in vivo after experimental and/or natural rhinovirus infections. Although it has long been recognized that the common cold is a potent trigger for symptoms of asthma, the mechanisms underlying the association between upper respiratory infection and increased lower airway obstruction remain obscure. The use of experimental infection of volunteers with or without respiratory allergies has enabled direct comparisons of common cold symptoms in these two groups. Furthermore, techniques such as bronchoalveolar lavage and segmental antigen challenge have been used to directly sample lower airway fluids and tissues during acute viral infection.

Antiallergic properties of antihistamines.

Histamine is a major mediator of the allergic reaction, and histamine H1-receptor antagonists have a long history of clinical efficacy in a variety of allergic disorders. The pathogenesis of allergic disease is complex, involving not only histamine and mast cell-derived tryptase, but also eosinophil and neutrophil derived mediators, cytokines, and intercellular adhesion molecules (ICAM-1). A number of "in vitro" and "in vivo" studies have been performed to assess the clinical effectiveness of antihistamines in inhibiting the allergen-induced inflammatory process in the skin and mucosa. In vitro human studies have shown that high concentration of second generation antihistamines can block inflammatory mediator release from basophils and mast cells, and reduce ICAM-1 expression in epithelial cell lines. In vivo studies have also shown an effect on the allergen-induced inflammatory reaction; both oral and intranasal antihistamines cause a reduction in nasal symptoms and inflammatory cell influx. Analysis of secretory fluids and tissues after challenge indicates that antihistamines interfere with mediator release. Recruitment of inflammatory cells to the site of the allergic insult is also disturbed by antihistamines of second-generation, suggesting that these drugs may inhibit upregulation of molecules involved in cell adhesion and migration, and perhaps they may interfere with the cytokine cascade through their ability of stabilizing mast cells and of limiting the incursion of inflammatory cells. This article reviews available human data on the antiallergic effects of antihistamines.

[Treatment of asthma using aerosols]. / Tratamiento del asma con aerosoles.

The vast majority of clinicians in Europe now prescribe beta-2 agonists as first-line therapy for patients with asthma. Inhaler devices may deliver rapidly acting (beta-2 sympathomimetics) and more slowly acting (anticholinergic) bronchodilator therapy as well as prophylactic medication (sodium cromoglycate and topical corticosteroids). The metered dose inhaler (MDI) is most often prescribed, but at least 50% of patients cannot use this device efficiently and 10-15% of those who can, develop an inefficient technique. The vast majority of those patients are able to use a single-dose dry power inhaler. Recent studies have shown that a multidose dry power system can be used by most patients and is preferred to the MDI by over two-thirds of patient. The large volume spacer systems have been shown to be as good as the MDI and nebulizer systems in the management of asthma, and they are easier to use than the MDI. Nebulisers are of value in chronic asthma in children who cannot use other delivery systems. The role of nebulisers for the domiciliary treatment of asthma in adults, however, remains controversial.

[Comparative study] of beclomethasone and budesonide, with same posology (400 micrograms every 12 hours), in the control of cortico-dependent intrinsic asthma]. / Estudio comparativo entre beclometasona y budesonida, a igual posología (400 micrograms cada 12 horas), en el control del asma intrínseco corticodependiente.

Inhaled steroids have rendered an undoubtful benefit in the control of airway inflammation of the asthmatic patients. Our objective was to compare clinical efficacy between budesonide (BUD) and beclomethasona depropionate (BDP), when administered at equal doses (800 micrograms/24 hours). A two ways crossed open clinical trial was designed. Thirty-three steroid dependent chronic asthmatic patients (18 females, 15 males) were included. Ages ranged from 29 to 73 years (mean = 52.5 +/- 11.7). All subjects suffered a severe asthma, with several years of activity (mean = 11.7 +/- 7.8), insufficiently controlled by inhaled steroids and bronchodilators, who required regular systemic steroids supply. The parameters compared were: the patients subjective symptoms punctuation (cough, expectoration, thoracic noises, exercise induce dyspnea and dyspnea at rest), salbutamol needs (number of inhalations/day), additional prednisone needs, sputum eosinophil counts, FEV1 measurement and inespecific bronchial reactivity control (PD20 methacoline). After a baseline week patients received one of the drugs for 6 weeks and, after a lavage week, the other drug was administered for another 6 weeks. All patients improved with both therapies. We got the following conclusions: 1) a significative decrease in salbutamol (p < 0.05-0.001) and prednisone needs (p < 0.05-0.001); 2) this decrease has been more important during BUD therapy, although without significative differences; 3) no significant variations in sputum eosinophils, FEV1 or bronchial reactivity were observed; 4) both drugs, when administered at equal doses, have probed to be equally effective in severe steroid dependent asthma control.

[Non-sedative antihistaminics in the treatment of chronic urticaria]. / Los antihistamínicos no sedantes en el tratamiento de la urticaria crónica.

Antihistamines are the drugs of choice in the symptomatic relief of chronic idiopathic urticaria; however, the usefulness of classic antihistamines has been limited by side effects. In the 1980s a new class of antihistamines has been developed that maintains effectiveness and produces less side effects (eg anticholinergic side effects, daytime sedation, etc). This review analyzes each of the new nonsedating antihistamines commercially available in Spain (astemizole, ebastine, cetirizine, loratadine and terfenadine) and evaluates its clinical efficacy and safety in the treatment of chronic idiopathic urticaria.

Determination of eosinophil rate after nasal provocation test in allergic rhinitis diagnosis.

After nasal provocation test in patients with allergic rhinitis, using the allergen they were sensitized to, we have observed: 1) an increase in the percentage of nasal eosinophils after 2, 3, 24 and 48 hours; 2) sneezes, mainly in the first 30 minutes; 3) nasal obstruction in the first three hours; 4) absence of rhinorrhea, but not in all the patients; and 5) no predominance of nasal, auricular and/or palatine pruritus at any time. When patients without rhinitis, or with allergic rhinitis were stimulated using a pneumoallergen they were not sensitized to, no significative increase in the nasal eosinophils percentage was found. No symptoms were observed either. So, we can conclude that nasal secretion samples, for eosinophilia percentage determination, should be taken from 2 to 48 hours after nasal provocation, and that the most frequent symptoms, which are probably related to cellular changes, are nasal obstruction and sneezes.

[Adverse reactions to food preservatives]. / Reacciones adversas a conservantes alimentarios.

We relate our experiences about the number of exacerbations that certain food preservatives such as sorbic acid, benzoic acid, sodium benzoate, metabisulfite and sodium nitrate can provoke in 62 patients affected with ASA-triad in steroid dependent intrinsic asthma with nasal polyps and acute bronchospasm caused by aspirin ingestion, and in 80 patients with chronic urticaria (C.U.) as well as the first assays of the possible usefulness of the HRT (Histamine release test automatized using whole blood) for the etiologic diagnosis process. In the cases of ASA-triad, and after the ingestion of aspirin (alternating with lactose in identical capsule), we consider the result as positive when the reduction of FEV1 is superior to 20% from its baseline value. Regarding the cases of C.U., the symptoms always exacerbate twice as much with the same substance within 24 hours of its administration. We have performed the HRT on 59 patients (14 with ASA-triad, 11 with steroid dependent intrinsic asthma; 20 with C.U. were negative to oral intake of analgesics/additives and 14 with C.U. showed positive results). Successive dilutions were incubated for 30 minutes using: pyrazolones, acetylsalicylate of lysine, sodium salicylate, sodium benzoate and 4-hydroxybenzoic acid which did not produce liberation of histamine in 100 controlled individuals. All the determinations were done in duplicate, considering positive those superior to 20% of the difference between total and basal histamine. We have not observed any significant descent of the FEV1 with benzoate and salicylate in our group of 62 patients with ASA-triad, nor any manifestations presented with sodium metabisulfite, sodium nitrate and sorbic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

[Immediate hypersensitivity to fruits and vegetables and pollenosis]. / Hipersensibilidad inmediata a frutas y verduras y polinosis.

Various works have been published, mainly by Scandinavian authors, in which a partial immunological identity has been found to exist between birch pollen and hazelnut and apple. However, our attention has been particularly drawn to the high proportion of people who are allergic to the apple and other fruits, which in some cases has been as much as 50 and 70%, and in whom an oral pruritus alone is considered sufficient a symptom of allergies to certain foodstuffs. However due to the fact that the incidence of allergies to fruit in patients who suffer from pollen-related allergies is much lower in this area, being in the order of 7%, and because of the absence of birch, we decided to carry out our investigation in the opposite direction to that chosen by the Scandinavian researchers. That is to say, from the point of view of patients suffering from allergies to fruits and vegetables (these being the foodstuffs which most frequently produce allergic reactions in our own particular environment) we decided to investigate: 1) which foodstuffs, according to clinical history, are most frequently responsible for producing allergies; 2) their possible co-existence with pollen related allergies; 3) which pollens are responsible for producing a higher rate of positive results in patients; 4) and finally, by means of the RAST inhibition technique, to discover it a cross-reactivity might exist between these pollens and the peanut or the pea. We chose 40 patients (19 females and 21 males), between the ages of 5 and 49 years old, whose clinical history showed beyond a shadow of a doubt that they displayed symptoms of immediate hypersensitivity after the ingestion of certain fruits and/or vegetables. The technique employed in the skin tests was that of scratch of the skin from the backs of the patients (in a prone position), systematically using fresh natural foodstuffs, and at the same time trying to ensure that they were of the same type as those that had originally produced the above-mentioned symptoms in the patients. On 193 occasions we carried out passive transfers (the Prausnitz-Küstner method or P.K.), and on 13 patients we carried out oral provocative tests (cases with positive scratch and uncertain clinical histories). With regard to the pollens, we used glycerine-based antigens of the type: Lolium perenne, Olea europeae, Artemisia vulgaris and Parietaria officinalis of Dome/Hollister-Stier, and the Bencard gramineae.(ABSTRACT TRUNCATED AT 400 WORDS)