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1.
Clin Exp Allergy ; 42(6): 966-75, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22909168

RESUMEN

BACKGROUND: The mould Alternaria alternata is a major elicitor of allergic asthma. Diagnosis and specific immunotherapy (SIT) of Alternaria allergy are often limited by the insufficient quality of natural mould extracts. OBJECTIVE: To investigate whether recombinant Alt a 1 can be used for reliable diagnosis of Alternaria alternata allergy and to develop a safe, non-allergenic vaccine for SIT of Alternaria allergy. METHODS: The qualitative sensitization profile of 80 Alternaria-allergic patients from Austria and Italy was investigated using an allergen micro-array and the amount of Alternaria-specific IgE directed to rAlt a 1 was quantified by ImmunoCAP measurements. Peptides spanning regions of predicted high surface accessibility of Alt a 1 were synthesized and tested for IgE reactivity and allergenic activity, using sera and basophils from allergic patients. Carrier-bound peptides were studied for their ability to induce IgG antibodies in rabbits which recognize Alt a 1 and inhibit allergic patients' IgE reactivity to Alt a 1. RESULTS: rAlt a 1 allowed diagnosis of Alternaria allergy in all tested patients, bound the vast majority (i.e. >95%) of Alternaria-specific IgE and elicited basophil activation already at a concentration of 0.1 ng/mL. Four non-allergenic peptides were synthesized which, after coupling to the carrier protein keyhole limpet hemocyanin, induced Alt a 1-specific IgG and inhibited allergic patients' IgE binding to Alt a 1. CONCLUSIONS AND CLINICAL RELEVANCE: rAlt a 1 is a highly allergenic molecule allowing sensitive diagnosis of Alternaria allergy. Carrier-bound non-allergenic Alt a 1 peptides are candidates for safe SIT of Alternaria allergy.


Asunto(s)
Alternaria/inmunología , Antígenos Fúngicos/inmunología , Vacunas Fúngicas/inmunología , Hipersensibilidad/diagnóstico , Hipersensibilidad/prevención & control , Péptidos/inmunología , Adolescente , Adulto , Animales , Anticuerpos Antifúngicos/sangre , Anticuerpos Antifúngicos/inmunología , Anticuerpos Antifúngicos/metabolismo , Especificidad de Anticuerpos/inmunología , Antígenos Fúngicos/química , Niño , Femenino , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina E/metabolismo , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Masculino , Unión Proteica/inmunología , Pliegue de Proteína , Multimerización de Proteína , Estructura Secundaria de Proteína , Conejos , Adulto Joven
2.
Clin Exp Allergy ; 40(3): 385-97, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20210812

RESUMEN

Allergen-specific immunotherapy (SIT) is the only specific and disease-modifying approach for the treatment of allergy but several disadvantages have limited its broad applicability. We argue that the majority of the possible disadvantages of SIT such as unwanted effects, poor efficacy and specificity as well as inconvenient application are related to the poor quality of natural allergen extracts, which are the active ingredients of all currently available allergy vaccines. Because of the progress made in the field of molecular allergen characterization, new allergy vaccines based on recombinant allergens, recombinant hypoallergenic allergen derivatives and allergen-derived T cell peptides have entered clinical testing and hold promise to reduce the side-effects and to increase the specificity as well as the efficacy of SIT. Here, we present a refined immunotherapy concept, which is based on the use of peptides derived from allergen surfaces that exhibit reduced, allergen-specific IgE as well as T cell reactivity. These peptides when fused to non-allergenic carriers give rise to allergen-specific protective IgG responses with T cell help from a non-allergenic carrier molecule. We summarize the experimental data demonstrating that such peptide vaccines can bypass allergen-specific IgE as well as T cell activation and may be administered at high doses without IgE- and T cell-mediated side-effects. Should these peptide vaccines prove efficacious and safe in clinical trials, it may become possible to develop convenient, safe and broadly applicable forms of SIT as true alternatives to symptomatic, drug-based allergy treatment.


Asunto(s)
Alérgenos/inmunología , Desensibilización Inmunológica/métodos , Hipersensibilidad/terapia , Inmunoglobulina E/inmunología , Péptidos/inmunología , Linfocitos T/inmunología , Vacunas/inmunología , Relación Dosis-Respuesta Inmunológica , Humanos , Hipersensibilidad/inmunología
3.
Clin Exp Allergy ; 40(4): 679-87, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20447079

RESUMEN

BACKGROUND: Allergy to fig fruit (Ficus carica) has been described in patients allergic to Ficus benjamina or rubber latex but may occur also in pollen-allergic patients. OBJECTIVE: To study the potential cross-reactivity between fig and taxonomically related fruits with the major birch pollen allergen Bet v 1. METHODS: One hundred and eighty-eight patients with or without birch pollen allergy were prick-to-prick tested with fig (F. carica), mulberry (Morus alba), jackfruit (Artocarpus heterophyllus; all family Moraceae) and other pollen-associated foods. Moraceae fruit extracts were separated by SDS-PAGE and tested with patient sera and polyclonal antisera against Mal d 1. Western blot inhibition was performed with Moraceae fruit extracts, birch pollen and recombinant Bet v 1. Putative Bet v 1 homologs in Moraceae fruits were analysed by liquid chromatography-ion trap mass spectrometry. RESULTS: Among 85 patients with isolated birch pollen allergy, 78% had a positive skin test to fresh fig, 10% to dried fig, 91% to mulberry, 91% to jackfruit, 77% to Rosaceae fruits and 83% to hazelnut. Sixty-six per cent of birch pollen-allergic patients positive for fig, reported symptoms after consumption of fresh figs, whereas dried figs were mostly well tolerated. In 60 patients with isolated Ficus benjamina sensitization, the reactivity rates to the same foods were 83-40-0-0-0-0%. None of 32 mugwort pollen-allergic patients reacted to Moraceae fruits. Rabbit anti-Mal d 1 and patient sera reacted to a 17 kDa band in all Moraceae extracts. IgE binding to these proteins was completely inhibited by birch pollen and rBet v 1. Mass spectrometry identified several peptides from the 17 kDa fig, mulberry and jackfruit allergen with respectively 60%, 56% and 76% homology to Bet v 1. CONCLUSION: Fig and other Moraceae fruits contain allergens homologous to Bet v 1 and represent clinically relevant birch pollen-associated foods.


Asunto(s)
Alérgenos/inmunología , Ficus/inmunología , Hipersensibilidad a los Alimentos , Frutas/inmunología , Moraceae/inmunología , Proteínas de Plantas/inmunología , Alérgenos/química , Secuencia de Aminoácidos , Antígenos de Plantas , Cromatografía Liquida , Reacciones Cruzadas , Hipersensibilidad a los Alimentos/etiología , Hipersensibilidad a los Alimentos/inmunología , Humanos , Hipersensibilidad/etiología , Hipersensibilidad/inmunología , Hipersensibilidad al Látex/etiología , Hipersensibilidad al Látex/inmunología , Espectrometría de Masas , Datos de Secuencia Molecular , Péptidos/química , Péptidos/inmunología , Proteínas de Plantas/química , Pruebas Cutáneas
4.
Int Arch Allergy Immunol ; 151(1): 17-27, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-19672093

RESUMEN

BACKGROUND: Allergen-specific subcutaneous immunotherapy (SCIT) is an antigen-specific therapy of IgE-mediated allergies. In the present study, we analyze the epitope specificities of antibody responses induced by SCIT with allergen extracts from pollen of trees belonging to the order Fagales (birch, alder, hazel) adsorbed onto aluminum hydroxide. METHODS: The IgE, IgG1-4 and IgA responses to defined recombinant allergens (birch pollen: Bet v 1; alder pollen: Aln g 1; hazel pollen: Cor a 1; apple: Mal d 1) as well as to Bet v 1-derived recombinant fragments and synthetic peptides were analyzed in sera from patients who had undergone SCIT for different periods of time. RESULTS: Long-term SCIT (>1 year; cumulative dose >1,000,000 SQ units) induced more pronounced IgG1, IgG2 and IgG4 responses to Bet v 1 and Bet v 1-related allergens according to the degree of sequence homology (Bet v 1>Aln g 1>Cor a 1>Mal d 1) than short-term SCIT (<1 year; cumulative dose <1,000,000 SQ units). In contrast to patients treated for <1 year, patients treated for >1 year mounted distinct IgG1, IgG2 and IgG4 responses against sequential Bet v 1 epitopes. No relevant allergen-specific IgA or IgG3 responses were induced by short- or long-term SCIT. Using a competitive ELISA assay, it could be shown that serum IgG from patients undergoing long-term SCIT inhibited IgE reactivity to Bet v 1 better than IgG from patients undergoing short-term SCIT. CONCLUSION: SCIT with allergen extracts adsorbed onto aluminum hydroxide induces IgG responses against new epitopes that block IgE binding and cross-react with structurally related allergens depending, among other factors, on duration of treatment and cumulative injected dose.


Asunto(s)
Alérgenos/administración & dosificación , Proteínas Recombinantes/administración & dosificación , Rinitis Alérgica Estacional/terapia , Adyuvantes Inmunológicos/farmacología , Adolescente , Adulto , Alérgenos/inmunología , Hidróxido de Aluminio/farmacología , Desensibilización Inmunológica , Mapeo Epitopo , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Inyecciones Subcutáneas , Masculino , Persona de Mediana Edad , Polen/inmunología , Proteínas Recombinantes/inmunología , Rinitis Alérgica Estacional/sangre , Rinitis Alérgica Estacional/inmunología , Adulto Joven
5.
Eur J Clin Invest ; 39(5): 429-36, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-19302561

RESUMEN

BACKGROUND: Commercial extracts used for diagnosis and treatment of allergy are currently prepared from natural allergen sources. The aim of this study was to analyse birch pollen allergen extracts produced for in vivo diagnosis of birch pollen allergy regarding their contents of individual birch pollen allergens (Bet v 1, Bet v 2 and Bet v 4). METHODS: Protein contents were measured and the allergen composition was analysed by immunoblotting using antibody probes specific for Bet v 1, Bet v 2 and Bet v 4 in birch pollen extracts from five manufacturers of allergen extracts. The contents of the major birch pollen allergen, Bet v 1, were quantified with a specific two-site binding enzyme-linked immunosorbent assay with nanogram sensitivity for Bet v 1. The biological activities of the allergen extracts were evaluated by skin prick testing in birch pollen allergic patients and compared with their sensitization profiles. RESULTS: A more than 10-fold variation regarding total protein contents (23.1-314 microg mL(-1)) and also regarding the amounts of the major birch pollen allergen, Bet v 1 (1.62-19.6 microg mL(-1)) was found. The highly cross-reactive Bet v 4 allergen was absent in three of the five tested extracts. Furthermore, varying skin test results were obtained in birch pollen allergic patients with the allergen extracts. CONCLUSIONS: Commercial birch pollen extracts exhibit a considerable variability regarding allergen contents and hence deliver varying in vivo test results. These problems might be overcome with recombinant allergen-based preparations.


Asunto(s)
Alérgenos/inmunología , Antígenos de Plantas/inmunología , Betula/inmunología , Hipersensibilidad/inmunología , Proteínas de Plantas/inmunología , Polen/inmunología , Adulto , Anciano , Alérgenos/biosíntesis , Femenino , Humanos , Immunoblotting , Masculino , Persona de Mediana Edad , Proteínas de Plantas/biosíntesis , Sensibilidad y Especificidad , Pruebas Cutáneas/métodos
6.
Clin Exp Allergy ; 38(8): 1400-8, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18564332

RESUMEN

BACKGROUND: The diagnosis and specific immunotherapy of allergy is currently performed with allergen extracts prepared from natural allergen sources. OBJECTIVE: To analyse commercial timothy grass pollen allergen extracts used for in vivo diagnosis regarding their qualitative and quantitative allergen composition and in vivo biological activity. METHODS: Antibodies specific for eight timothy grass pollen allergens (Phl p 1, Phl p 2, Phl p 4, Phl p 5, Phl p 6, Phl p 7, Phl p 12, Phl p 13) were used to detect these allergens in timothy grass pollen extracts from four manufacturers by immunoblotting. ELISA assays were developed and used to quantify the three major allergens (Phl p 1, Phl p 2, Phl p 5) in the extracts. The magnitude of skin responses to the four extracts was studied by skin prick testing in 10 grass pollen-allergic patients. RESULTS: The allergen extracts showed broad variations in protein compositions and amounts (24.1-197.7 microg/mL extract). Several allergens could not be detected in certain extracts or appeared degraded. A considerable variability regarding the contents of major allergens was found (Phl p 1: 32-384 ng/mL; Phl p 2: 1128-6530 ng/mL, Phl p 5: 40-793 ng/mL). Heterogeneous skin test results were obtained with the extracts in grass pollen-allergic patients. CONCLUSIONS: Timothy grass pollen extracts from different manufacturers exhibit a considerable heterogeneity regarding the presence of individual allergens and hence yield varying in vivo test results. Problems related to the use of natural grass pollen allergen extracts may be circumvented by using defined recombinant grass pollen allergens.


Asunto(s)
Alergia e Inmunología/normas , Antígenos de Plantas/análisis , Phleum/química , Extractos Vegetales/química , Polen/química , Antígenos de Plantas/inmunología , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Humanos , Immunoblotting , Phleum/inmunología , Extractos Vegetales/uso terapéutico , Polen/inmunología , Rinitis Alérgica Estacional/diagnóstico , Rinitis Alérgica Estacional/inmunología , Rinitis Alérgica Estacional/terapia , Pruebas Cutáneas/métodos , Pruebas Cutáneas/normas
7.
Clin Exp Allergy ; 38(10): 1688-96, 2008 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18754759

RESUMEN

BACKGROUND: Beech and oak pollen are potential allergen sources with a world-wide distribution. OBJECTIVE: We aimed to characterize the allergen profile of beech and oak pollen and to study cross-reactivities with birch and grass pollen allergens. METHODS: Sera from tree pollen-allergic patients with evidence for beech and oak pollen sensitization from Basel, Switzerland, (n=23) and sera from birch pollen-allergic patients from Vienna, Austria, (n=26) were compared in immunoblot experiments for IgE reactivity to birch (Betula pendula syn. verrucosa), beech (Fagus sylvatica) and oak (Quercus alba) pollen allergens. Subsequently, beech and oak pollen allergens were characterized by IgE inhibition experiments with purified recombinant and natural allergens and with allergen-specific antibody probes. Birch-, beech- and oak pollen-specific IgE levels were determined by ELISA. RESULTS: Beech and oak pollen contain allergens that cross-react with the birch pollen allergens Bet v 1, Bet v 2 and Bet v 4 and with the berberine bridge enzyme-like allergen Phl p 4 from timothy grass pollen. Sera from Swiss and Austrian patients exhibited similar IgE reactivity profiles to birch, beech and oak pollen extracts. IgE levels to beech and oak pollen allergens were lower than those to birch pollen allergens. CONCLUSION: IgE reactivity to beech pollen is mainly due to cross-reactivity with birch pollen allergens, and a Phl p 4-like molecule represented another predominant IgE-reactive structure in oak pollen. The characterization of beech and oak pollen allergens and their cross-reactivity is important for the diagnosis and treatment of beech and oak pollen allergy.


Asunto(s)
Alérgenos/inmunología , Betula/inmunología , Fagus/inmunología , Hipersensibilidad/inmunología , Inmunoglobulina E/inmunología , Polen/inmunología , Reacciones Antígeno-Anticuerpo , Austria , Reacciones Cruzadas/inmunología , Humanos , Hipersensibilidad/diagnóstico , Inmunoglobulina E/sangre , Quercus/inmunología , Suiza
8.
Clin Exp Allergy ; 38(7): 1192-202, 2008 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-18462454

RESUMEN

BACKGROUND: Patients with multiple sensitizations require alternative forms of treatment, as the efficacy of conventional immunotherapy is unsatisfactory. OBJECTIVE: In the present study, we sought to compare the efficacy of a subcutaneously (s.c.) and a mucosally applied polyvalent vaccine to reduce allergic immune responses within airway and lung tissues. METHODS: Female BALB/c mice were intraperitoneally immunized with recombinant (r)Bet v 1, rPhl p 1 and rPhl p 5, followed by an aerosol challenge of birch and phleum pollen extract. For tolerance induction, either a mixture of the immunodominant peptides or a hybrid peptide of the respective antigens was s.c. injected or intranasally applied before poly-sensitization. RESULTS: Mucosal but not systemic pre-treatment with poly-peptides led to significant suppression of eosinophils and IL-5 production in bronchoalveolar lavages, as well as IL-5, IL-4, IL-13 and eotaxin levels in lung cell cultures. Lung histology showed a clear reduction of cellular infiltration and mucus production only in intranasally pre-treated mice. In accordance, also the systemic immune response, characterized by IgE-dependent basophil degranulation and IL-4 levels in vitro, was significantly reduced by mucosal antigen application, but only marginally influenced by subcutaneous pre-treatment. Both treatment routes led to up-regulated CTLA4 expression in splenocytes, whereas only after mucosal pre-treatment Foxp3 expression levels were enhanced in lung CD3(+) T cells. Furthermore, intranasal but not subcutaneous application of the peptides enhanced IL-10 levels in the lungs, indicating regulatory mechanisms operating in local tolerance induction. CONCLUSION: Mucosal application of peptides is superior to systemic application in preventing both local and systemic poly-allergic T helper2 immune responses, suggesting mucosal tolerance induction as an attractive strategy for the primary and secondary prevention of allergic multi-sensitization and lung pathology.


Asunto(s)
Alérgenos/administración & dosificación , Desensibilización Inmunológica , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Inflamación/inmunología , Pulmón/inmunología , Péptidos/administración & dosificación , Alérgenos/inmunología , Animales , Basófilos/inmunología , Basófilos/metabolismo , Degranulación de la Célula , Citocinas/biosíntesis , Citocinas/inmunología , Eosinófilos/inmunología , Eosinófilos/metabolismo , Femenino , Hipersensibilidad/patología , Hipersensibilidad/prevención & control , Inmunoglobulina G/sangre , Inflamación/metabolismo , Inflamación/prevención & control , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Mucosa Nasal/inmunología , Péptidos/inmunología , Polen/inmunología , Ratas , Sistema Respiratorio/inmunología , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo
9.
Allergy ; 63(3): 310-26, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18269676

RESUMEN

Allergen extracts have been used for diagnosis and treatment of allergy for around 100 years. During the second half of 20th century, the notion increasingly gained foothold that accurate standardization of such extracts is of great importance for improvement of their quality. As a consequence, manufacturers have implemented extensive protocols for standardization and quality control. These protocols have overall IgE-binding potencies as their focus. Unfortunately, each company is using their own in-house reference materials and their own unique units to express potencies. This does not facilitate comparison of different products. During the last decades, most major allergens of relevant allergen sources have been identified and it has been established that effective immunotherapy requires certain minimum quantities of these allergens to be present in the administered maintenance dose. Therefore, the idea developed to introduce major allergens measurements into standardization protocols. Such protocols based on mass units of major allergen, quantify the active ingredients of the treatment and will at the same time allow comparison of competitor products. In 2001, an EU funded project, the CREATE project, was started to support introduction of major allergen based standardization. The aim of the project was to evaluate the use of recombinant allergens as reference materials and of ELISA assays for major allergen measurements. This paper gives an overview of the achievements of the CREATE project.


Asunto(s)
Alérgenos/clasificación , Guías como Asunto , Hipersensibilidad/diagnóstico , Proteínas Recombinantes , Estudios de Validación como Asunto , Cromatografía Líquida de Alta Presión/normas , Desensibilización Inmunológica/normas , Ensayo de Inmunoadsorción Enzimática/normas , Europa (Continente) , Femenino , Humanos , Masculino , Espectrometría de Masas/normas , Proteínas Recombinantes/normas , Estándares de Referencia , Valores de Referencia , Sensibilidad y Especificidad , Análisis Espectral/normas , Organización Mundial de la Salud
10.
FEBS Lett ; 261(1): 106-8, 1990 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-1968399

RESUMEN

Allicin is shown to be a specific inhibitor of the acetyl-CoA synthetases from plants, yeast and mammals. The bacterial acetyl-CoA-forming system, consisting of acetate kinase and phosphotransacetylase, was inhibited too. Non-specific interaction with sulfhydryl-groups could be excluded in experiments with dithioerythritol and p-hydroxymercuribenzoate. Binding of allicin to the enzyme is non-covalent and reversible. [14C]-Acetate incorporation into fatty acids of isolated plastids was inhibited by allicin with an I50-value lower than 10 microM. Other enzymes of the fatty acid synthesis sequence were not affected, as was shown using precursors other than acetate.


Asunto(s)
Acetato CoA Ligasa/antagonistas & inhibidores , Coenzima A Ligasas/antagonistas & inhibidores , Acetato CoA Ligasa/metabolismo , Acetatos/metabolismo , Animales , Bacterias/enzimología , Bovinos , Cloroplastos/enzimología , Disulfuros , Relación Dosis-Respuesta a Droga , Ácidos Grasos/biosíntesis , Miocardio/enzimología , Ácidos Sulfínicos/metabolismo , Ácidos Sulfínicos/farmacología , Levaduras/enzimología
11.
FEBS Lett ; 414(1): 129-34, 1997 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-9305746

RESUMEN

In further substantiating the novel mevalonate-independent pathway for isoprenoid biosynthesis, which generates isopentenyl diphosphate (IPP) via 1-deoxy-D-xylulose-5-phosphate, labeling experiments with 1-[2H(1)]deoxy-D-xylulose were performed with various higher plants and algae: efficient incorporation was observed into isoprene emitted by Populus, Chelidonium, and Salix, into the phytol moiety of chlorophylls in a red alga (Cyanidium), in two green algae (Scenedesmus, Chlamydomonas), and a higher plant (Lemna). By contrast, 13C-mevalonate applied was incorporated into isoprene and phytol to a much lower extent or not at all. This demonstrates that this '1-deoxy-D-xylulose-5-phosphate pathway' for biosynthesis of plastidic isoprenoids is widely distributed in photosynthetic organisms.


Asunto(s)
Butadienos/metabolismo , Eucariontes/metabolismo , Hemiterpenos , Magnoliopsida/metabolismo , Pentanos , Fitol/metabolismo , Xilulosa/análogos & derivados , Isótopos de Carbono , Chlorophyta/metabolismo , Deuterio , Glicósidos/metabolismo , Espectroscopía de Resonancia Magnética , Ácido Mevalónico/metabolismo , Estructura Molecular , Compuestos Organofosforados/metabolismo , Rhodophyta/metabolismo , Esteroles/biosíntesis , Xilulosa/metabolismo
13.
Artículo en Inglés | MEDLINE | ID: mdl-21337238

RESUMEN

Biogenic amines in wine may impair sensory wine quality and cause adverse health effects in susceptible individuals. In this study, histamine and other biogenic amines were determined by HPLC after amine derivatisation to dansyl chloride conjugates in 100 selected high-quality red wines made from seven different cultivars. Amine levels varied considerably between different wines. The most abundant amines were putrescine (median = 19.4 mg l(-1), range = 2.9-122), histamine (7.2 mg l(-1), 0.5-26.9), and tyramine (3.5 mg l(-1), 1.1-10.7), whereas lower levels were found for isoamylamine (median = 0.25 mg l(-1)), phenylethylamine (0.16 mg l(-1)), cadaverine (0.58 mg l(-1)), spermidine (1.8 mg l(-1)) and tryptamine (0.06 mg l(-1)). Positive correlations were observed between isoamylamine and phenylethylamine, and between histamine, putrescine and tyramine levels. Amine concentrations were similar in all wine cultivars except Pinot noir and St. Laurent wines, which showed significantly higher tryptamine and cadaverine levels. The results indicate that levels of histamine and other biogenic amines may vary considerably between red wines independent of grape variety and that high amounts can also be found in high-rated wines. Adopting a legal histamine threshold level of 10 mg l(-1) in the European Union, as formerly introduced in other countries, would have excluded 34% of the investigated wines from the market.


Asunto(s)
Aminas Biogénicas/análisis , Contaminación de Alimentos , Histamina/análisis , Vino/análisis , Métodos Analíticos de la Preparación de la Muestra , Aminas Biogénicas/química , Cromatografía Líquida de Alta Presión , Compuestos de Dansilo/química , Colorantes Fluorescentes/química , Histamina/química , Límite de Detección , Putrescina/análisis , Putrescina/química , Control de Calidad , Reproducibilidad de los Resultados , Especificidad de la Especie , Tiramina/análisis , Tiramina/química
14.
Clin Exp Allergy ; 34(8): 1251-8, 2004 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-15298566

RESUMEN

BACKGROUND: Ornamental Ficus benjamina (FB) has been recognized as a new indoor allergen. Little is known about the prevalence in moderately exposed subjects and the proposed association with fruit and Hevea latex hypersensitivity. OBJECTIVE: To study the prevalence of FB sensitization and the relationship with Hevea latex allergy, to identify cross-reacting fruits, and to characterize the responsible allergens. METHODS: A skin prick test solution prepared from FB latex (200 microg/mL) was included in our routine screening programme for suspect inhalant allergy. Patients reacting with the FB extract were further skin tested with exotic fruits by the prick-to-prick method. Inhibition of fig and FB CAP by FB latex, fig (Ficus carica), kiwi, the thiolproteases ficin and papain, Hevea latex and rHev b 6.02 (hevein) was performed in selected patients. RESULTS: Of 2662 patients with a positive skin test to any aeroallergen, 66 (2.5%) reacted with FB. Ten patients showed isolated sensitization to FB. Although FB-positive subjects were more often co-sensitized to Hevea latex than FB-negative (10.6% vs 3.8%, P< 0.01), nearly 90% tested negative for Hevea latex. Sensitization to FB was specifically associated with positive skin tests to fresh fig (83%), dried fig (37%), kiwi fruit (28%), papaya (22%), avocado (19%), banana (15%), and pineapple (10%) (n = 54). Clinical reactions were reported mainly from fresh and dried fig and kiwi (47%, 60%, and 64%, respectively, of skin test-positive patients), including seven patients with systemic reactions (urticaria, angiooedema, asthma). CAP to fig in 11 patients with clinical fruit allergy was inhibited on average by 87% by FB latex, 89% by fresh fig, 80% by dried fig, 38% by kiwi (100 microg/mL each), and by 59% and 44% by ficin and papain (50 microg/mL), respectively. No inhibition was obtained with Hevea latex and rHev b 6.02. CAP to FB was inhibited on average by 95% by FB, 60% by fresh fig, 41% by ficin, 29% by papain, and less than 7% by rubber latex allergens. CONCLUSIONS: Sensitization to FB latex is found in 2.5% of atopic individuals and mostly occurs independently of Hevea latex allergy. Sensitization is commonly associated with allergic reactions to figs and other tropical fruits ('Ficus-fruit syndrome'). This cross-reactivity is mediated at least in part by thiolproteases.


Asunto(s)
Alérgenos/inmunología , Ficus , Hipersensibilidad/diagnóstico , Actinidia , Adulto , Ananas , Carica , Reacciones Cruzadas , Hipersensibilidad a los Alimentos/complicaciones , Hipersensibilidad a los Alimentos/inmunología , Hevea , Humanos , Hipersensibilidad/inmunología , Hipersensibilidad al Látex/complicaciones , Hipersensibilidad al Látex/inmunología , Tamizaje Masivo , Musa , Prevalencia , Pruebas Cutáneas , Síndrome
15.
Plant J ; 22(1): 39-50, 2000 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10792819

RESUMEN

The principal supply of carbon precursors for fatty acid synthesis in leaf tissue has been a much debated topic, with some experiments suggesting a direct supply from the C3 products of photosynthetic carbon fixation and colleagues suggesting the utilization of free acetate (for which concentrations in leaves in the range of 0.05-1.4 mM have been reported). To address this issue we first reassessed the in vivo rate of fatty acid synthesis using a new method, that of [13C]carbon dioxide labeling of intact Arabidopsis plants with the subsequent analysis of fatty acids by gas chromatography-mass spectrometry (GC-MS). This method gave an average value of 2.3 mmoles carbon atoms h-1 mg chlorophyll-1 for photosynthetic tissues. The method was extended by isotopic dilution analysis to measure the rate of fatty acid synthesis in the dark. There was negligible fatty acid synthesis (< 5% of the rate in the light) in the dark. In addition, the method allowed an estimate of the absolute rate of fatty acid degradation of about 4% of the total fatty acid content per day. With the in vivo rate of fatty acid synthesis in the light defined, if the bulk tissue acetate concentration available for fatty acid synthesis is 1 mM, this acetate pool can sustain fatty acid synthesis for approximately 60 min. When the leaves of Arabidopsis, barley and pea were given a 5 min pulse of [14C]carbon dioxide, the label rapidly appeared in fatty acids with a lag phase of less than 2-3 min. Continuous labeling with [14C]carbon dioxide, for up to 1 h, showed a similar result. Furthermore, 14C-label in free acetate was less than 5% of that in fatty acids. In conclusion, these data suggest that either the bulk pool of acetate is not involved in fatty acid synthesis or the concentration of acetate must be less than 0.05 mM under strong illumination.


Asunto(s)
Ácidos Grasos/biosíntesis , Hojas de la Planta/metabolismo , Ácido Acético/metabolismo , Acetilcoenzima A/metabolismo , Arabidopsis/metabolismo , Arabidopsis/efectos de la radiación , Carbono/metabolismo , Dióxido de Carbono/metabolismo , Isótopos de Carbono , Radioisótopos de Carbono , Oscuridad , Cromatografía de Gases y Espectrometría de Masas , Cinética , Luz , Fotosíntesis
16.
Clin Exp Allergy ; 34(3): 460-9, 2004 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-15005742

RESUMEN

BACKGROUND: IgE antibodies against carbohydrate epitopes have been identified recently as a major cause of in vitro double positivity to honeybee (HB) and vespid venom in patients with stinging-insect allergy. As these antibodies possibly have low clinical relevance they may be misleading in the diagnosis of venom allergy. OBJECTIVE: To confirm the role of carbohydrate epitopes in double positivity and to locate the responsible glycoallergens in HB and yellow jacket (YJ) venom by western blot. METHODS: Immunoblot inhibition using HB venom, YJ venom and two glycoprotein sources displaying 1-3-fucosylated N-glycans (i.e. oilseed rape (OSR) pollen, and the synthetic neo-glycoprotein fucosylated/xylosylated N-glycans from bromelain coupled to bovine serum albumin (MUXF-BSA)) as inhibitors were performed with sera from 15 double-positive patients with stinging-insect allergy. Additionally, reactivity with blotted hymenoptera venoms of a carbohydrate-specific rabbit antiserum against OSR pollen was investigated. RESULTS: Major venom glycoallergens binding with carbohydrate-specific human IgE and rabbit IgG were detected in HB venom at 42 (hyaluronidase (HYA)), 46, 65 and 95 kDa, and in YJ venom at 38 and 43 kDa (HYA). Antibody binding to these allergens was completely lost after periodate treatment. Glycans of HB phospholipase were bound by patients' IgE only after protein denaturation. In 10 of the 15 patients the reactivity was with the second venom because of carbohydrates alone. The high-molecular-weight glycoallergens identified in HB venom probably correspond to similar proteins described earlier, including allergens B and C. The 38-kDa YJ allergen might represent a homologue of V mac 3. CONCLUSIONS: The data confirm the proposed role of carbohydrate-specific IgE in double positivity to HB and YJ venom and shed new light on some previously described minor hymenoptera allergens of uncertain clinical significance. The consideration of carbohydrate-specific IgE may allow to discriminate between patients with potentially relevant and patients with non-relevant double sensitization.


Asunto(s)
Alérgenos/análisis , Venenos de Abeja/química , Glicoproteínas/análisis , Hipersensibilidad Inmediata/inmunología , Venenos de Avispas/química , Adolescente , Adulto , Niño , Reacciones Cruzadas , Femenino , Humanos , Hipersensibilidad Inmediata/tratamiento farmacológico , Immunoblotting/métodos , Inmunoglobulina E/inmunología , Masculino , Persona de Mediana Edad , Ácido Peryódico/uso terapéutico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
17.
Allergy ; 58(5): 448-51, 2003 May.
Artículo en Inglés | MEDLINE | ID: mdl-12752335

RESUMEN

BACKGROUND: Allergy to houseflies is rare. We report a case of respiratory allergy from occupational exposure to houseflies in a farmer. CASE REPORT: A 30 year-old female farmer with a long-standing history of grass pollen allergy observed for 2 years rhino-conjunctivitis and mild asthma when entering livestock stables and barns. Allergy retesting revealed sensitization to various pollens but not to animal danders. Houseflies (Musca domestica) occurring on the farm in great quantity were suspected by the farmer herself as the causative agent. RESULTS: Skin prick testing with housefly was positive in the patient and negative in four controls. Experimental radioallergosorbant test was class 3 positive. Sensitization to house dust mite, storage mites and cockroach was not detectable. Western blots with housefly extracts revealed immunoglobulin E (IgE)-binding to bands of 70, 50, and approximately 16 kDa. Tropomyosin in the housefly extract (35 kDa) was recognized by a tropomyosin reference serum but not by the patient. In enzyme-linked immunosorbent assay (ELISA) inhibition assays using housefly as the solid phase, IgE-binding of the patient was inhibited by 75% by M. domestica and by 44% by the closely related lesser housefly (Fannia canicularis), but not by extracts from blowfly (Lucilia spp.), fruit fly (Drosophila spp.), horsefly (Haematopota pluvialis) and mosquito (Culex pipiens). The IgE-binding of the tropomyosin control serum was inhibited by 60-80% by all species. CONCLUSIONS: In accordance with previous reports, this case demonstrates that respiratory sensitization to insects may be highly specific. According to ELISA inhibition, cross-sensitization in the present case was restricted to species of the family of true flies (Muscidae).


Asunto(s)
Moscas Domésticas , Enfermedades Profesionales/etiología , Exposición Profesional/efectos adversos , Hipersensibilidad Respiratoria/etiología , Adulto , Animales , Femenino , Humanos
18.
J Am Acad Dermatol ; 35(3 Pt 1): 377-80, 1996 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-8784272

RESUMEN

BACKGROUND: Contact dermatitis from artificial nails made from self-curing acrylic resins is occasionally reported. Recently, UV light-cured products introducing new acrylics have become available. OBJECTIVE: Our purpose was to identify relevant allergens in commercial light-curing products by patch tests and to evaluate the efficacy of "hypoallergenic" products by inclusion into the test series. METHODS: Patients wearing photobonded acrylic nails who had perionychial and subonychial eczema were patch tested with an acrylate battery and "hypoallergenic" commercial products. RESULTS: Triethyleneglycol dimethacrylate, hydroxyfunctional methacrylates, and (meth)-acrylated urethanes proved to be relevant allergens in photobonded nail preparations. Methacrylated epoxy resin sensitization was not observed. All "hypoallergenic" products provoked positive reactions. CONCLUSION: The omission of irritant methacrylic acid in UV-curable products does not reduce the high sensitizing potential of new acrylates. In contrast to the manufacturers' declarations, all "hypoallergenic" products continue to include acrylate functional monomers and therefore continue to cause allergic sensitization.


Asunto(s)
Resinas Acrílicas/efectos adversos , Cosméticos/efectos adversos , Dermatitis Alérgica por Contacto/etiología , Enfermedades de la Uña/inducido químicamente , Uñas , Acrilatos/efectos adversos , Resinas Acrílicas/aislamiento & purificación , Resinas Acrílicas/efectos de la radiación , Alérgenos/efectos adversos , Alérgenos/aislamiento & purificación , Bisfenol A Glicidil Metacrilato/efectos adversos , Resinas Compuestas/efectos adversos , Cosméticos/efectos de la radiación , Eccema/inducido químicamente , Resinas Epoxi , Femenino , Dedos , Humanos , Irritantes/efectos adversos , Metacrilatos/efectos adversos , Pruebas del Parche , Polietilenglicoles/efectos adversos , Ácidos Polimetacrílicos/efectos adversos , Poliuretanos/efectos adversos , Rayos Ultravioleta
19.
Hautarzt ; 51(4): 266-9, 2000 Apr.
Artículo en Alemán | MEDLINE | ID: mdl-10810664

RESUMEN

We report on a child with pili trianguli et canaliculi. This hair shaft abnormality belongs to a heterogeneous group of diseases which are included under the synonym uncombable hair. The diagnosis was confirmed by scanning electron microscopy, revealing hair shafts with a characteristic longitudinal groove. In addition the girl suffered from atopic eczema and tooth anomalies. Our findings suggest that this disorder could represent a tricho-odontal subtype of ectodermal dysplasia.


Asunto(s)
Dermatitis Atópica/genética , Displasia Ectodérmica/genética , Cabello/anomalías , Anomalías Dentarias/genética , Biotina/administración & dosificación , Preescolar , Dermatitis Atópica/diagnóstico , Displasia Ectodérmica/diagnóstico , Femenino , Cabello/patología , Humanos , Microscopía Electrónica de Rastreo , Anomalías Dentarias/diagnóstico
20.
Allergy ; 55(10): 923-30, 2000 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11030372

RESUMEN

BACKGROUND: The role of ash (Fraxinus excelsior) pollen as a cause of spring pollinosis in central Europe has received little attention. It is not clear whether ash pollen is a primary cause of sensitization or whether it is implicated through cross-sensitization to other pollens. METHODS: Over a 22-month period, ash pollen was included in a screening series for inhalant allergies. Pollen data were documented from 1976 through 1999. The frequency of IgE-binding to the ash-specific allergen Fra e 1 and pollen panallergens, respectively, was compared by Western blot between mono- (n = 6), oligo- (n = 16), and polysensitized (n=25) patients. RESULTS: Of 5,416 consecutive patients sensitized to any pollen, 920 (17.6%) had a positive skin prick test to ash. Total pollen counts varied extensively between years (229-5,351) as did peak concentrations (23-837 grains/m3/24 h). Western blotting revealed Fra e 1 sensitization in 100% of monosensitized, 93% of oligosensitized, but only 44% of polysensitized patients. IgE against profilins (Fra e 2), Ca-binding proteins (Fra e 3), and carbohydrate epitopes in the three groups was found in 0/0/17%, 0/19/31%, and 32/72/60%, respectively. At least 50% of sera from patients with Fra e 1 sensitization did not bind with the protein in Western blots under reducing conditions. CONCLUSIONS: Ash pollen should be considered a relevant factor and distinct entity in spring pollinosis. In all, only 20% of positive skin tests to ash appear to result from cross-sensitization to pollen panallergens.


Asunto(s)
Alérgenos/inmunología , Proteínas Contráctiles , Polen/inmunología , Rinitis Alérgica Estacional/inmunología , Alérgenos/metabolismo , Austria , Western Blotting , Proteínas de Unión al Calcio/metabolismo , Carbohidratos/inmunología , Epítopos/metabolismo , Humanos , Inmunoglobulina E/inmunología , Inmunoglobulina E/metabolismo , Magnoliopsida/inmunología , Proteínas de Microfilamentos/metabolismo , Profilinas , Prueba de Radioalergoadsorción , Rinitis Alérgica Estacional/etiología , Pruebas Cutáneas , Árboles/inmunología
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