Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 53
Filtrar
Mais filtros










Filtros aplicados
Base de dados
Intervalo de ano de publicação
2.
J. investig. allergol. clin. immunol ; 32(5): 367-374, 2022. ilus, tab
Artigo em Inglês | IBECS | ID: ibc-212732

RESUMO

Background: Platanus acerifolia (London plane tree) is a deciduous tree of the Platanaceae family. Sensitization to this plant varies withgeography. Madrid, located in central Spain, has one of the highest London plane tree pollen concentration levels on the Iberian Peninsula.Objectives: We evaluated both the clinical characteristics and the molecular sensitization pattern of patients with allergy to London planetree pollen in the region of Madrid.Patients and Methods: Thirty-eight patients allergic to London plane tree pollen were selected according to their clinical symptoms andpositive results in skin prick testing and/or specific IgE determination. Serum was collected, and allergen components were evaluatedusing immunodetection techniques as well as ImmunoCAP. The IgE-binding proteins detected were identified and characterized usingmass spectrometry.Results: Analysis of serum samples from allergic patients revealed 9 IgE-binding bands in London plane tree pollen extract. Among these,the 45-kDa protein, which corresponded to Pla a 2, was detected in 76.3% of patients. However, the 18-kDa (Pla a 1) and 9-kDa (Pla a 3)bands were detected in 44.7% and 23.7% of sera, respectively. These results were confirmed using purified proteins. Characterization ofthe allergen revealed the 27-kDa protein to be glutathione-S-transferase.Conclusions: The molecular profile of patients sensitized to London plane tree pollen differs from that reported in studies from otherlocations. In the population we studied, the prevalence of Pla a 2 was higher than that of Pla a 1 and Pla a 3. In addition, the minorallergen previously referred to as Pla a 4 was characterized as glutathione-S-transferase. (AU)


Antecedentes: Platanus acerifolia es un árbol de hoja caduca de la familia Platanaceae. La sensibilización frente a esta planta varía enfunción de la zona geográfica. Madrid, ubicada en el centro de España, tiene uno de los mayores niveles de concentración de polen deeste árbol en la Península Ibérica.Objetivo: Evaluar las características clínicas y los patrones moleculares de sensibilización en pacientes con alergia al plátano de sombraen la región de Madrid.Pacientes y Métodos: Treinta y ocho pacientes alérgicos al polen del plátano de sombra fueron seleccionados de acuerdo con los síntomasclínicos, pruebas cutáneas positivas y/o IgE específica. El suero se recogió y se evaluaron los componentes alérgicos mediante técnicas deinmunodetección, así como ImmunoCAP. Las proteínas que unían IgE fueron identificadas y caracterizadas por espectrometría de masas.Resultados: El análisis de los sueros de los pacientes alérgicos reveló 9 bandas que captaban IgE en los extractos de polen de plátano desombra. Entre estas, la proteína de 45 kDa, correspondiente a Pla a 2, se detectó en el 76,3% de los pacientes. Sin embargo, las bandasde 18 kDa (Pla a 1) y 9 kDa (Pla a 3) fueron reconocidas en el 44,7% y 27,3%, respectivamente. Estos resultados se confirmaron usandoproteínas purificadas. La caracterización de los alérgenos identificó la proteína de 27 kDa como una glutatión S-transferasa.Conclusiones: El perfil molecular de los pacientes sensibilizados al polen del plátano de sombra varía respecto al descrito en estudios deotras localizaciones. Nuestra población muestra una mayor prevalencia de Pla a 2 comparado con Pla a 1 y Pla a 3. Además, el alérgenominoritario previamente denominado Pla a 4 fue caracterizado como una glutatión-S-transferasa. (AU)


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Antígenos de Plantas/análise , Hipersensibilidade/diagnóstico , Magnoliopsida/imunologia , Pólen/imunologia , Alérgenos/imunologia , Hipersensibilidade/epidemiologia , Prevalência , Espanha/epidemiologia , Hipersensibilidade Imediata , Imunoglobulina E/imunologia , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática
3.
J. investig. allergol. clin. immunol ; 32(5): 383-392, 2022. ilus, graf
Artigo em Inglês | IBECS | ID: ibc-212734

RESUMO

Background: Immediate and delayed-type hypersensitivity reactions to pet-borne allergens are common in atopic diseases. In atopic dermatitis (AD), controversy surrounds the contribution to the disease of cross-reactivity to self-proteins. Human cystatin A and the cat allergen Fel d 3 belong to the cystatins, an evolutionary conserved protein family. The objective of the present study was to assess crossreactivity between mammalian cystatins and to analyze T-cell responses to cystatin in AD patients sensitized to pet dander. Methods: cDNA coding for dog cystatin was cloned from dog skin. Sera from 245 patients with IgE-mediated sensitization to cat and dog dander were tested for IgE binding to recombinantly expressed feline, canine, and human cystatin. Of these, 141 were also diagnosed with AD. Results: Cystatin-specific IgE was detected in 36 patients (14.7%), of whom 19 were considerably affected by AD. Within the AD patients, 9 had measurable IgE against all 3 cystatins. Cystatin-sensitized AD patients did not differ from non–cystatin-sensitized patients in terms of disease severity, age, or total IgE levels. T-cell cytokine measurements showed elevated IL-4 levels after stimulation with feline and human cystatin. Conclusion: The humoral response suggests that in addition to Fel d 3, the homologous protein from dog might play a role in allergy. Furthermore, human cystatin appears to be capable of driving a type 2 immune response in sensitized AD patients and may therefore be considered a so-called autoallergen, as proposed for other evolutionary conserved proteins. (AU)


Antecedentes: Las reacciones de hipersensibilidad de tipo inmediato y retardado a los alérgenos que están en las mascotas son comunes en las enfermedades atópicas. En este estudio, en pacientes con dermatitis atopica (DA), se analiza la reactividad cruzada con las autoproteínas y su contribución a la enfermedad. Tanto la cistatina A humana como el alérgeno felino Fel d 3 pertenecen a la familia de las cistatinas, una familia de proteínas conservadas evolutivamente. El objetivo del presente estudio fue evaluar la reactividad cruzada entre las cistatinas de mamíferos y analizar la respuestas de las células T a la cistatina en pacientes con DA sensibilizados a la caspa de las mascotas. Métodos: El ADNc que codifica la cistatina de perro se clonó a partir de piel de perro. Se analizaron sueros de 245 pacientes con sensibilización por IgE a la caspa de gato y perro para determinar la unión de IgE a cistatina felina, canina y humana expresada de forma recombinante, respectivamente. De estos 245 pacientes, 141 fueron diagnosticados de DA. Resultados: Se detectó IgE específica frente a cistatina en el 14,7% (36) de los pacientes, de los cuales 19 padecían DA. Dentro de los pacientes con DA, 9 tenían IgE medible contra las tres cistatinas. Los pacientes con DA sensibilizados frente a cistatina no difirieron de los pacientes no sensibilizados con cistatina en términos de gravedad de la enfermedad, edad o niveles totales de IgE. El análisis de citocinas de células T reveló niveles elevados de IL-4 después de la estimulación con cistatina felina y humana. Conclusión: La respuesta humoral sugiere que, además de Fel d 3, la proteína homóloga de perro también podría desempeñar un papel en la alergia. Además, la cistatina humana parece ser capaz de promover una respuesta inmune de tipo 2 en pacientes con DA sensibilizados y, por lo tanto, puede considerarse un autoalérgeno, como se ha propuesto para otras proteínas conservadas evolutivamente. (AU)


Assuntos
Humanos , Animais , Gatos , Cães , Dermatite Atópica/etiologia , Animais de Estimação , Apresentação Cruzada , Cistatinas/imunologia , Linfócitos T/imunologia , Dermatite Atópica/imunologia , Ensaio de Imunoadsorção Enzimática , Eletroforese em Gel de Poliacrilamida
4.
Allergol. immunopatol ; 48(6): 597-602, nov.-dic. 2020. graf, tab
Artigo em Inglês | IBECS | ID: ibc-199248

RESUMO

INTRODUCTION AND OBJECTIVES: Moths are a significant source of indoor and outdoor aeroallergens. High prevalence of IgE-mediated sensitization was demonstrated in a group of patients with allergic respiratory diseases. There are no studies on adult stage of these moth species allergens involved in allergic respiratory reactions - the aim of this study. MATERIAL AND METHODS: 36 participants were included in an experimental study, submitted to skin prick test with Bombyx mori wing extract and six other common allergens, as well as Western blot analysis with incubated nitrocellulose membrane impregnated with silkworm moth extract and human IgE-antibody. The participants were divided into 3 groups: 1) 21 allergic patients whose skin prick test was positive to Bombyx mori wing extract, 2) eight allergic patients whose skin prick test was positive to mite and negative to Bombyx mori extract 3) seven negative non-allergic subjects. RESULTS: Among the 21 participants from group 1, 19 serum samples reacted to Bombyx mori extract by Western blot. All of them reacted to a protein at 80 kDa and five other proteins (66, 50, 45, 37 and 30 kDa) were identified in more than 50% of the individuals tested, considered as major allergenic proteins. Sera from seven out of eight patients sensitized to house dust mite demonstrated IgE-reactivity to Bombyx mori extract by Western blot analysis. Serum samples from healthy participants did not react at all. CONCLUSION: Six major reactive proteins by immunoblot analysis from moth’s wings sensitized patients can be potential allergens. The one at 80 kDa is the major protein, seen in all IgE-reactive patients from group 1 and in none from group 2, yet to be identified. Future studies should be conducted to better characterize these proteins


No disponible


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Hipersensibilidade Respiratória/imunologia , Bombyx/química , Proteínas/análise , Alérgenos/análise , Bombyx/patogenicidade , Western Blotting , Imunoglobulina E/imunologia , Eletroforese em Gel de Poliacrilamida , Valores de Referência
5.
Allergol. immunopatol ; 48(4): 348-354, jul.-ago. 2020. tab, graf, ilus
Artigo em Inglês | IBECS | ID: ibc-199719

RESUMO

INTRODUCTION AND OBJECTIVES: This study aimed to compare the effects of different biotinylation methods on the performance characteristics of allergen-specific IgE detection. MATERIALS AND METHODS: The Gal d 6 gene was cloned into the pAN6/pAC6 vector, resulting in rGal d 6-Bio/Bio-rGal d 6 vector. The fusion protein was expressed in Escherichia coli AVB101 and simultaneously biotinylated in a site-specific manner. The Gal d 6 gene was amplified via PCR and cloned into the pET-28a vector and transformed into E. coli BL21 and purified via Ni-NTA, followed by chemical biotinylation using Sulfo-NHS-LC-Biotin. Twenty-eight patients allergic to hen's egg white were examined for sensitization against egg yolk. An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed to detect allergen-specific IgE. RESULTS: rGal d 6, Bio-rGal d 6, and rGal d 6 were prepared using different biotin binding modes to detect allergen-specific IgE. rGal d 6-Bio (Kd = 0.6154) and Bio-rGal d 6 (Kd = 0.6698) had a markedly better detection performance than rGal d 6 (Kd = 28.93), and the rGal d 6-Bio had a better detection performance in small-volume serum samples. CONCLUSIONS: rGal d 6-Bio improved the sensitivity for the detection of allergen-specific IgE


No disponible


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Hipersensibilidade Imediata/diagnóstico , Biotinilação , Anafilaxia/diagnóstico , Hipersensibilidade Alimentar/diagnóstico , Sensibilidade e Especificidade , Eletroforese em Gel de Poliacrilamida , Immunoblotting
6.
J. investig. allergol. clin. immunol ; 30(6): 421-429, 2020. tab, graf
Artigo em Inglês | IBECS | ID: ibc-202594

RESUMO

BACKGROUND: Mites are the most prevalent source of indoor allergens. The present study used a component-resolved diagnosis (CRD) approach to investigate the mite-specific IgE sensitization profile for Dermatophagoides pteronyssinus and Blomia tropicalis. We also assessed the performance of a commercially available CRD approach in patients with severe allergic rhinitis. METHODS: We selected 63 consecutive patients with dual sensitization to D pteronyssinus and B tropicalis and persistent severe rhinitis according to the ARIA guidelines. We performed skin prick tests with standardized extracts and determined specific serum IgE to both mites, along with serum specific IgE to Der p 1, Der p 2, Der p 23, Der p 10, and Blo t 5. RESULTS: Fifty-eight and 59 patients had positive sIgE to the whole extracts of D pteronyssinus and B tropicalis, respectively. While 91.67% of patients were sensitized to specific IgE to Der p 1, Der p 2, and/or Der p 23, specific IgE to Blo t 5 (≥0.3 ISU-E) was not detected in most of the serum samples (55%). CONCLUSIONS: Although the combination panel of the commercially available major allergens Der p 1, Der p 2, and Der p 23 identified more than 90% of the D pteronyssinus-allergic patients, Blo t 5 performed somewhat poorly in those sensitized to B tropicalis. Improvements in CRD and further research concerning the prevalence and clinical relevance of serodominant allergens are needed to achieve a genuine molecular diagnosis, as well as patient-centered mite allergy-specific immunotherapy


INTRODUCCIÓN: Los ácaros son los alérgenos de interior más prevalentes. El presente estudio investiga el perfil de sensibilización a Dermatophagoides pteronyssinus y Blomia tropicalis, así como el rendimiento del diagnóstico por componentes (CRD) disponible comercialmente en pacientes con rinitis alérgica grave persistente. MATERIAL Y MÉTODOS: Seleccionamos 63 pacientes con rinitis grave persistente (Guía ARIA) con sensibilización dual a D. pteronyssinus y B. tropicalis. Se realizaron pruebas cutáneas en prick con extractos estandarizados, IgE sérica específica a ambos ácaros además de IgE específica a alérgenos individuales Der p 1, Der p 2, Der p 23, Der p 10 y Blo t 5. RESULTADOS: Cincuenta y ocho y 59 pacientes presentaron IgE específica positiva a extractos crudos de D. pteronyssinus y B. tropicalis, respectivamente. Aunque el 91,67% mostraron sensibilización a Der p 1, Der p 2 y/o Der p 23, Blo t 5 (≥0,3 ISU-E) no fue detectado en la mayoría (55%) de las muestras estudiadas. CONCLUSIONES: Aunque la combinación de alérgenos principales Der p 1, Der p 2 Der p 23, pudo identificar más del 90% de los pacientes sensibilizados a D. pteronyssinus, Blo t 5 presentó un rendimiento diagnóstico muy limitado para aquellos sensibilizados a B. tropicalis. Conocer la prevalencia y relevancia clínica de los alérgenos acarianos serodominantes en cada territorio contribuiría a una mejor identificación de sensibilizaciones genuinas en la era de la medicina de precisión


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto Jovem , Adulto , Rinite Alérgica/diagnóstico , Rinite Alérgica/etiologia , Dermatophagoides pteronyssinus , Hipersensibilidade Imediata , Antígenos de Dermatophagoides/imunologia , Índice de Gravidade de Doença , Testes Cutâneos , Eletroforese em Gel de Poliacrilamida , Western Blotting
7.
Nutr. clín. diet. hosp ; 40(3): 153-161, 2020. ilus, graf
Artigo em Inglês | IBECS | ID: ibc-201599

RESUMO

OBJECTIVES: To determine the effects of W100E-Leptin in a streptozotocin-induced diabetic mice model (effects in the body weight, fasting serum glucose and glucose tolerance). METHODS: Intraperitoneal W100E-Leptin application at 1 mg/kg for 13 days. We used 3 experimental groups (n=6). Group 1: Diabetes + W100E-Leptin (intraperitoneal administration), Group 2: Diabetes + buffer (vehicle) and Group 3: Healthy control + buffer (vehicle). RESULTS: We determined the effects of W100E on the behavior of the mice, more active, more hair and a tendency to gain body weight. We did not observe any hypoglycemic effect of W100E-Leptin on serum glucose levels in the tests we performed. CONCLUSIONS: These results show us the need to characterize the effects of this hormone in diabetes. We will continue with the characterization of the change that is generated in the protein regulation caused by W100E-Leptin in the diabetes, to propose this hormone as an adjunct against diabetes


No disponible


Assuntos
Animais , Camundongos , Diabetes Mellitus Experimental/metabolismo , Leptina/metabolismo , Glicemia/análise , Eletroforese em Gel de Poliacrilamida , Teste de Tolerância a Glucose
9.
Int. microbiol ; 22(2): 289-296, jun. 2019. graf, tab
Artigo em Inglês | IBECS | ID: ibc-184835

RESUMO

Wheat gluten proteins are decisive for the industrial properties of flour, so alterations resulting from grain infection with Fusarium graminearum produce changes in the glutenin content that affect the baking properties. This work analyzes the high-molecular-weight glutenin changes from wheat flour with different degrees of F. graminearum infection at field, since these proteins are determinant for the quality properties of flour. Wheat cultivars-on field trials-infected with F. graminearum isolates of diverse aggressiveness showed severity values between 9.1 and 42.58% and thousand kernel weight values between 28.12 and 32.33 g. Negative correlations between severity and protein content and positive correlations between yield and protein content were observed, employing reversed-phase high-performance liquid chromatography and polyacrylamide gel electrophoresis. Furthermore, the protein signal changes were in agreement for both methodological approaches. Also, the degree of disease observed and the protein changes on infected wheat cultivars varied in relation with the aggressiveness of the isolate responsible for the infection. The principal component analysis showed a close arrangement among protein values obtained by HPLC. For each cultivar, two principal components were obtained, which explained 80.85%, 88.48%, and 93.33% of the total variance (cultivars Sy200, AGP Fast, and Klein Tigre respectively). To our knowledge, the approaches employed for the analysis of protein changes according to the degree of disease, as well as the thorough statistical analysis, are novel for the study of Fusarium Head Blight


No disponible


Assuntos
Triticum/microbiologia , Farinha/microbiologia , Grão Comestível/microbiologia , Proteínas de Grãos/análise , Fusarium/patogenicidade , Fungos/patogenicidade , Contaminação de Alimentos/análise , Glutens/análise , Fusarium/crescimento & desenvolvimento , Cromatografia Líquida de Alta Pressão/métodos , Eletroforese em Gel de Poliacrilamida/métodos
10.
Rev. iberoam. micol ; 36(2): 66-71, abr.-jun. 2019. tab, ilus, graf
Artigo em Inglês | IBECS | ID: ibc-185478

RESUMO

Background: Members of the Pleosporaceae family are known as important sources of airborne allergens which are responsible for asthma and allergic diseases. Aims: The purpose of this study was to investigate the gene profiling and expression pattern of Alt a 1 in Alternaria alternata and other members of the Pleosporaceae family including Stemphylium botryosum, Ulocladium chartarum, Curvularia lunata, Cladosporium cladosporioides, and Epicoccum nigrum. Methods: Alternaria alternata and related genera were cultured on Czapek-Dox broth medium at 25°C for 21 days. The presence of Alt a 1 was assessed in fungal culture filtrates by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then confirmed by immunoblot analysis. Real-time PCR was carried out for quantitation of the Alt a 1 gene encoding corresponding protein at the transcriptional level using cDNA prepared from fungal RNA. Results: SDS-PAGE showed protein bands ranging from 14 to 100 kDa. A 14kDa band corresponding to Alt a 1 was present in A. alternata, S. botryosum and U. chartarum. The gene expression of Alt a 1 was reported in A. alternata and some other related genera. The Ct mean value recorded for A. alternata strains ranged from 24.70 to 27.84 while it was in the range 23.62-32.09 for other related taxa. No apparent transcription or expression was revealed in C. cladosporioides. Conclusions: The presence and efficient expression of Alt a 1 gene in A. alternata and other related taxa indicate that Alt a 1 protein is a major component of the secretory machinery of Pleosporaceae family members, and it may play a crucial role in its allergenicity


Antecedentes: Los miembros de la familia Pleosporaceae son una fuente importante de alérgenos aéreos causantes de asma y enfermedades alérgicas. Objetivos El objetivo de este trabajo fue estudiar el perfil de expresión génica de la proteína Alt a 1 en Alternaria alternata y otros miembros de la familia Pleosporaceae, entre las cuales pueden citarse Stemphylium botryosum, Ulocladium chartarum, Curvularia lunata, Cladosporium cladosporioides y Epicoccum nigrum. Métodos: Alternaria alternata y otros géneros relacionados se cultivaron en caldo Czapek-Dox a 25°C durante 21 días. La existencia de Alt a 1 en los filtrados de los cultivos se evaluó mediante electroforesis en gel de poliacrilamida con dodecilsulfato sódico (SDS-PAGE) para después confirmarla mediante inmunotransferencia. Se realizó RCP en tiempo real para la cuantificación de la transcripción del gen responsable (Alt a 1) utilizando ADNc a partir del ARN del hongo. Resultados: Mediante SDS-PAGE se visualizaron bandas de proteínas de 14 a 100 kDa. La banda de 14 kDa, correspondiente a Alt a 1, estaba presente en A. alternata, S. botryosum y U. chartarum. Se detectó expresión génica de Alt a 1 en A. alternata y otros géneros relacionados. El valor medio de Ct registrado en los aislamientos de A. alternata varió entre 24,70 y 27,84; en otros taxones cercanos, el intervalo estuvo entre 23,62 y 32,09. No se detectó transcripción o expresión en C. cladosporioides. Conclusiones: La presencia del gen Alt a 1 y su expresión en A. alternata y otros taxones próximos indica que la proteína Alt a 1 es uno de los componentes principales del mecanismo secretorio de los miembros de la familia Pleosporaceae y puede desempeñar un papel fundamental en su capacidad alergénica


Assuntos
Humanos , Alérgenos/genética , Alternaria/genética , Antígenos de Fungos/genética , Ascomicetos/genética , Proteínas Fúngicas/genética , Perfilação da Expressão Gênica , Alérgenos/análise , Alternaria/imunologia , Antígenos de Fungos/análise , Ascomicetos/imunologia , Eletroforese em Gel de Poliacrilamida , Proteínas Fúngicas/análise , Immunoblotting
16.
Clin. transl. oncol. (Print) ; 16(2): 153-157, feb. 2014. tab, ilus
Artigo em Inglês | IBECS | ID: ibc-127718

RESUMO

PURPOSE: To construct a protein catalogue of malignant pleural effusion from lung adenocarcinoma patients and to screen the potential candidates of biomarkers for diagnostic value in human lung adenocarcinoma. METHOD: Five malignant pleural effusion samples of lung adenocarcinoma patients were collected from January 2009 to September. A composite sample was analyzed using shotgun strategy. Pleural effusion samples were separated by means of SDS-PAGE. Proteomic analysis was performed by 1D-LC-MS/MS, and then the proteins were identified using SEQUEST software and protein database search. RESULTS: Among 230 unique proteins, 123 proteins were identified with higher confidence levels (at least two unique peptide sequences matched). Most of these proteins have been reported in plasma. However, there are 7 proteins, including JUP protein, suprabasin, annexin A2, transforming growth factor-beta-induced protein ig-h3 (βig-h3), V-set and immunoglobulin domain-containing protein 4 precursor, ifapsoriasin 2 and actin, cytoplasmic 1 have not been reported in serum. CONCLUSIONS: Seven proteins may represent potential candidates of biomarkers. Annexin A2 is of special interest since it may play a role in the regulation of intercellular adhesion and cell proliferation (AU)


No disponible


Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Adenocarcinoma/metabolismo , Neoplasias Pulmonares/metabolismo , Derrame Pleural Maligno/metabolismo , Proteoma/análise , Proteômica/métodos , Biomarcadores Tumorais/análise , Adenocarcinoma/complicações , Adenocarcinoma/patologia , Eletroforese em Gel de Poliacrilamida , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/patologia , Derrame Pleural Maligno/patologia , Biomarcadores Tumorais/metabolismo
17.
J. investig. allergol. clin. immunol ; 23(3): 159-167, mayo-jun. 2013. tab, ilus
Artigo em Inglês | IBECS | ID: ibc-114859

RESUMO

Antecedentes: La globalización de la industria alimentaria proporciona la exposición a nuevos pescados no domésticos y se hace necesaria la identificación de los alérgenos potenciales para diagnosticar las reacciones alérgicas. Objetivo: El objetivo de este estudio fue estudiar las proteínas fijadoras de IgE que constituyen los alérgenos de la perca del Nilo (L. niloticus). Métodos: Mediante electroforesis 2D en gel se separaron las proteínas del músculo del L. niloticus y G. morhua y se enfrentaron al suero de 12 pacientes con historia de reacción inmediata a pescado, así como al suero de pacientes atópicos y controles sanos. Las proteínas reactivas a IgE fueron identificadas mediante espectrofotometría de masas. Results: En los resultados, el paciente mostraba un índice bajo de fijación de IgE a parvalbúminas, sin embargo mostraba fijación de IgE a 8 alérgenos diferentes a la parvalbúmina de L. niloticus y 5 a la G. morhua. Observamos una sensibilización cruzada de 7/12 (58%) de los individuos alérgicos a pescado a la enolasa-3 del L. niloticus, mientras que 11/12 (92%) de los pacientes estaban sensibilizados a la enolasa-3 del G. morhua. Conclusión: La identificación de los alérgenos especie-específicos o de la sensibilización individual podría en el futuro mejorar las estrategias de evitación (AU)


Background: Globalization of the food industry has led to widespread exposure to new nondomestic fish species; therefore, identification of potential allergens is necessary in order to diagnose allergic reactions. Objective: Contact with a patient who was allergic to Nile perch (Lates niloticus) prompted us to investigate the immunoglobulin (Ig) E– reactive proteins that could be allergens of this species. Methods: 2D gel electrophoresis was used to separate the muscle proteins of L niloticus and Gadus morhua. Immunoblotting was performed with sera from 12 patients with a history of immediate-type allergic reaction to fish and from atopic and nonatopic controls. IgE-reactive proteins were detected and identifi ed using mass spectrometry. Results: The index patient had low levels of IgE binding to parvalbumins. However, 8 putative allergens other than parvalbumin from L niloticus and 5 from G morhua were identified. Further investigation revealed cross-sensitivity to enolase 3 from L niloticus in 7 of the 12 fish-allergic individuals (58%), whereas 11 of the 12 patients (92%) were sensitized to enolase 3 from G morhua. However, atopic control patients were also sensitized to enolase 3 from L niloticus and G morhua. Conclusion: Identification of species-specific allergens and individual sensitization could help us to improve avoidance strategies (AU)


Assuntos
Humanos , Proteínas de Peixes/efeitos adversos , Alérgenos/isolamento & purificação , Hipersensibilidade Alimentar/prevenção & controle , Proteínas de Peixes/imunologia , Fosfoglucomutase/imunologia , Fosfopiruvato Hidratase/imunologia , Proteínas de Ligação ao Cálcio/imunologia , Eletroforese em Gel de Poliacrilamida , Creatina Quinase/imunologia , Testes Cutâneos , Ensaio de Imunoadsorção Enzimática , Western Blotting
18.
J. investig. allergol. clin. immunol ; 23(3): 176-182, mayo-jun. 2013. ilus, tab
Artigo em Inglês | IBECS | ID: ibc-114861

RESUMO

Antecedentes: La larva de Protophormia terraenovae, utilizada como cebo vivo para la pesca, es capaz de producir reacciones alérgicas en el 7% de la población de pescadores de agua dulce de Cáceres, según las observaciones previas de nuestro grupo. Objetivo: Identificar el patrón de alérgenos en P. terranovae y otros Califóridos. Materiales y métodos: Los extractos de P. terraenovae, Calliphora vomitoria, Lucilia sericata y Lumbricus terrestris se sometieron a técnicas de SDS-PAGE e IgE-immunoblotting utilizando sueros individuales de 24 pacientes sensibilizados a P. terraenovae. Se realizaron también técnicas de ELISA e IgE-immunoblotting inhibition para la identificación de posibles alérgenos comunes entre dichas especies. Resultados: 15 pacientes reconocieron una banda entorno a los 15.3 kDa frente al extracto de P. terraenovae, además de otros 2 alérgenos de 22.8 and 69kDa. Con C. vomitoria, se observaron 5 bandas de 73, 46, 40, 28 y 14 kDa. Con L. sericata se observaron 2 alérgenos mayores de 73 y 14 kDa. Usando L terrestris, 13 pacientes reconocieron un alérgeno de unos 15.5 kDa. Los estudios de inhibición IgE demostraron la presencia de reactividad cruzada inmunológica principalmente entre L. terrestris y P. terraenovae. Conclusiones: La larva de P. terraenovae parece tener alérgenos especie-específi cos y alérgenos compartidos con C. vomitoria y L. sericata. Se ha observado una importante reactividad cruzada inmunológica entre P. terraenovae y L. terrestris. Un alérgeno entre los 15-16 kDa podría ser uno de los responsables de este fenómeno (AU)


Background: Our group previously found that up to 7% of amateur anglers in Caceres, Spain may be allergic to the larvae of Protophormia terraenovae (order Diptera, family Calliphoridae) used as live bait for fishing. Objective: To identify the pattern of major allergens in P terraenovae and other species of Calliphoridae. Materials and Methods: Extracts of P terraenovae, Calliphora vomitoria, Lucilia sericata and Lumbricus terrestris were characterized using sodium dodecyl sulfate polyacrylamide gel electrophoresis and IgE-immunoblotting techniques in individual sera from 24 patients with a positive skin test result and/or specific IgE determination (enzyme-linked immunosorbent assay [ELISA]) to P terraenovae. ELISA and IgE-immunoblotting inhibition studies were also performed to identify potential cross-reactive allergens between these species. Results: IgE-immunoblotting with P terraenovae showed a band of 15.3 kDa recognized by 15 patients, in addition to 2 further allergens of 22.8 kDa and 69 kDa. For C vomitoria, 5 bands of 73, 46, 40, 28, and 14 kDa were observed. For L sericata, 2 major allergens of 73 kDa and 14 kDa were observed. In the case of L terrestris, IgE from 13 patients recognized 1 allergen of around 15.5 kDa. IgE-immunoblotting and ELISA inhibition revealed the presence of cross-reactivity, mainly between L terrestris and P terraenovae. Conclusions: P terraenovae appears to have species-specific allergens and allergens shared with C vomitoria and L sericata. Striking immunological cross-reactivity was observed between P terraenovae and L terrestris. An allergen of 15-16 kDa could be involved in this phenomenon (AU)


Assuntos
Animais , Oligoquetos/imunologia , Alérgenos/análise , Alérgenos/química , Alérgenos , Dessensibilização Imunológica , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade/diagnóstico , Hipersensibilidade/imunologia , Peixes/imunologia , Alérgenos/imunologia , Eletroforese em Gel de Poliacrilamida/métodos , Eletroforese em Gel de Poliacrilamida/normas , Eletroforese em Gel de Poliacrilamida , Western Blotting/métodos , Western Blotting/normas , Western Blotting , Ensaio de Imunoadsorção Enzimática/instrumentação , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática
19.
Int. microbiol ; 13(3): 113-121, sept. 2010. ilus, tab
Artigo em Inglês | IBECS | ID: ibc-84635

RESUMO

The prophage Lv1, harbored by a vaginal Lactobacillus jensenii isolate, was induced by several different anticancer, antimicrobial, and antiseptic agents, suggesting that they contribute to the adverse vaginal effects associated with their therapeutic use. Of special interest with respect to its novelty was the inducing effect of nonoxynol-9, a non-ionic detergent commonly used as a spermicide. The Lv1 genome consists of a 38,934-bp dsDNA molecule with cohesive ends, in which 48 ORFs were recognized, and is organized into functional modules. Lv1 belongs to the family Siphoviridae and, more precisely, to the proposed Sfi21-like genus. The capsid-tail junction of the Lv1 virions is fragile such that most particles become disrupted, suggesting that the virus is defective and thus unable to generate fertile progeny. However, genome analysis did not provide evidence of the defective nature of the prophage, other than the finding that its genome is shorter than those of other, related, phages. Further analysis indicated that prophage Lv1 suffered deletions in its right half to the extent that it no longer fulfill the minimum packaging limits, thereby generating the observed unstable particles (AU)


No disponible


Assuntos
Humanos , Feminino , Genoma Viral , Lactobacillus/virologia , Prófagos/isolamento & purificação , Ativação Viral , Anti-Infecciosos Locais/metabolismo , Antineoplásicos/metabolismo , DNA Viral/química , DNA Viral/genética , Eletroforese em Gel de Poliacrilamida , Lactobacillus/isolamento & purificação , Microscopia Eletrônica de Transmissão , Prófagos/classificação , Prófagos/genética , Prófagos/fisiologia , Vagina/microbiologia , Replicação Viral
20.
Int. microbiol ; 13(2): 67-77, jun. 2010. ilus, tab
Artigo em Inglês | IBECS | ID: ibc-84631

RESUMO

Seven maritime Antarctic lakes located on Byers Peninsula (Livingston Island, South Shetland Islands) were surveyed to determine the relationship between planktonic bacterial community composition and environmental features. Specifically, the extent to which factors other than low temperature determine the composition of bacterioplankton assemblages of maritime Antarctic lakes was evaluated. Both deep and shallow lakes in the central plateau of the Peninsula, as well as a coastal lake, were studied in order to fully account for the environmental heterogeneity of the Peninsula's lakes. The results showed that shallow coastal lakes display eutrophic conditions, mainly due to the influence of marine animals, whereas plateau lakes are generally deeper and most are oligotrophic, with very limited inputs of nutrients and organic matter. Meso-eutrophic shallow lakes are also present on the Peninsula; they contain microbial mats and a higher trophic status because of the biologically mediated active nutrient release from the sediments. Diversity studies of the lakes' planktonic bacterial communities using molecular techniques showed that bacterial diversity is lower in eutrophic than in oligotrophic lakes. The former also differed in community composition with respect to dominant taxa. Multivariate statistical analyses of environmental data yielded the same clustering of lakes as obtained based on the DGGE band pattern after DNA extraction and amplification of 16S rRNA gene fragments. Thus, even in extremely cold lakes, the bacterial community composition parallels other environmental factors, such as those related to trophic status. This correspondence is not only mediated by the influence of marine fauna but also by processes including sediment and ice melting dynamics. The bacterial community can therefore be considered to be equally representative as environmental abiotic variables in demonstrating the environmental heterogeneity among maritime Antarctic lakes (AU)


No disponible


Assuntos
Bactérias/classificação , Bactérias/genética , Biodiversidade , Microbiologia da Água , Impressões Digitais de DNA , DNA Ribossômico/genética , DNA Bacteriano/genética , Eletroforese em Gel de Poliacrilamida , Geografia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...