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The historical stone heritage that we inherit must be passed on to future generations, not only in the same conditions that we found it but, if possible, in better ones. Construction also demands better and more durable materials, often stone. The protection of these materials requires knowledge of the types of rocks and their physical properties. The characterization of these properties is often standardized to ensure the quality and reproducibility of the protocols. These must be approved by entities whose purpose is to improve the quality and competitiveness of companies and to protect the environment. Standardized water absorption tests could be envisaged to test the effectiveness of certain coatings in protecting natural stone against water penetration, but we found that some steps of these protocols neglect any surface modification of the stones, and hence may not be completely effective when a hydrophilic protective coating (i.e., graphene oxide) is present. In this work, we analyze the UNE 13755/2008 standard for water absorption and propose alternative steps to adapt the norm for use with coated stones. The properties of coated stones may invalidate the interpretation of the results if the standard protocol is applied as is, so here we pay special attention to the characteristics of the coating applied, the type of water used for the test, the materials used, and the intrinsic heterogeneity of the specimens.
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Objective. The aims of this study are to compare 2 origins of adipose-derived mesenchymal stem cells (MSCs) (omentum and subcutaneous) from 2 pathologies (morbid obesity and cancer) vs healthy donors. Adipose tissue has revealed to be the ideal MSC source. However, in developing adipose-derived stem cells (ASCs) for clinical use, it is important to consider the effects of different fat depots and also the effect of donor variability. Methods. We isolated and characterized the membrane markers and differentiation capacities of ASCs obtained from patients with these diseases and different origin. During the culture period, we further analysed the cells' proliferation capacity in an in vitro assay as well as their secretome. Results. Adipose-derived stem cells isolated from obese and cancer patients have mesenchymal phenotype and similar cell proliferation as ASCs derived from healthy donors, some higher in cells derived from subcutaneous fat. However, cells from these 2 types of patients do not have the same differentiation potential, especially in cancer patients from omentum, and exhibit distinct secretion of both pro-inflammatory and regulatory cytokines, which could explain the differences in use due to origin as well as pathology associated with the donor. Conclusion. Subcutaneous and omentum ASCs are slightly different; omentum generates fewer cells but with greater anti-inflammatory capacity. Adipose-derived stem cells from patients with either obesity or cancer are slightly altered, which limits their therapeutic properties.
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Células-Tronco Mesenquimais , Neoplasias , Obesidade Mórbida , Tecido Adiposo , Humanos , Células-Tronco Mesenquimais/metabolismo , Omento , Gordura SubcutâneaRESUMO
Most concrete studies are concentrated on mechanical properties especially strength properties either directly or indirectly (fresh and durability properties). Hence, the ratio of split tensile strength to compressive strength plays a vital role in defining the concrete properties. In this review, the impact of design parameters on the strength ratio of various grades of Self-Compacting Concrete (SCC) with recycled aggregate is assessed. The design parameters considered for the study are Water to Cement (W/C) ratio, Water to Binder (W/B) ratio, Total Aggregates to Cement (TA/C) ratio, Fine Aggregate to Coarse Aggregate (FA/CA) ratio, Water to Solid (W/S) ratio in percentage, superplasticizer (SP) content (kg/cu.m), replacement percentage of recycled coarse aggregates (RCA), replacement percentage of recycled fine aggregates (RFA), fresh density and loading area of the specimen. It is observed that the strength ratio of SCC with recycled aggregates is affected by design parameters.
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INTRODUCTION: The COVID-19 pandemic has resulted in unparalleled changes to patient care, including the suspension of cancer surgery. Concerns regarding COVID-19-related risks to patients and healthcare workers with the re-introduction of major complex minimally invasive and open surgery have been raised. This study examines the COVID-19 related risks to patients and healthcare workers following the re-introduction of major oesophago-gastric (EG) surgery. PATIENTS AND METHODS: This was an international, multi-centre, observational study of consecutive patients treated by open and minimally invasive oesophagectomy and gastrectomy for malignant or benign disease. Patients were recruited from nine European centres serving regions with a high population incidence of COVID-19 between 1 May and 1 July 2020. The primary endpoint was 30-day COVID-19-related mortality. All staff involved in the operative care of patients were invited to complete a health-related survey to assess the incidence of COVID-19 in this group. RESULTS: In total, 158 patients were included in the study (71 oesophagectomy, 82 gastrectomy). Overall, 87 patients (57%) underwent MIS (59 oesophagectomy, 28 gastrectomy). A total of 403 staff were eligible for inclusion, of whom 313 (78%) completed the health survey. Approaches to mitigate against the risks of COVID-19 for patients and staff varied amongst centres. No patients developed COVID-19 in the post-operative period. Two healthcare workers developed self-limiting COVID-19. CONCLUSIONS: Precautions to minimise the risk of COVID-19 infection have enabled the safe re-introduction of minimally invasive and open EG surgery for both patients and staff. Further studies are necessary to determine the minimum requirements for mitigations against COVID-19.
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COVID-19 , Pandemias , Pessoal de Saúde , Humanos , Procedimentos Cirúrgicos Minimamente Invasivos , SARS-CoV-2RESUMO
BACKGROUND: The best alimentary and biliopancreatic limb (BPL) lengths in the Roux-en-Y gastric bypass (RYGB) still remain unclear. The aim of this study was to compare the effect of a BPL of 70 vs 120 cm, with a constant AL of 150 cm on long-term weight loss, remission of comorbidities, and supplementation needs after RYGB. PATIENTS AND METHODS: A prospective randomized study of morbidly obese patients undergoing RYGB was performed. Patients were randomized into two groups: those patients undergoing RYGB with a BPL of 70 cm (BPL 70 cm) and those ones undergoing RYGB with a BPL of 120 cm (BPL 120 cm). BMI, excess BMI loss (EBMIL), remission of comorbidities and specific vitamin and mineral supplementation needs at 1, 2, and 5 years were analyzed. RESULTS: Two hundred fifty-three patients were included in each group. There were no significant differences in BMI, EBMIL and the remission of diabetes mellitus, hypertension, and dyslipidemia between groups at 1, 2, and 5 years after surgery. Patients from group BPL 120 cm required greater specific supplementation of vitamin B12, folic acid, and vitamin A during all the follow-up. CONCLUSION: A RYGB with 120 cm BPL does not achieve greater weight loss or remission of comorbidities than a RYGB with 70 cm BPL but is associated with greater deficiencies of vitamin B12, vitamin A, and folic acid. TRIAL REGISTRATION: ClinicalTrials.gov Identifier NCT03607305. https://clinicaltrials.gov/.
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Deficiência de Vitaminas/tratamento farmacológico , Comorbidade , Derivação Gástrica/métodos , Vitaminas/administração & dosagem , Redução de Peso , Adulto , Deficiência de Vitaminas/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
Objetivos: Conocer los factores asociados a sobrepeso y obesidad en niños atendidos en el Hospital San Bartolomé. 2005-2010. Material y métodos: la muestra estuvo constituida por 149 niños de 9 a 11 años de edad atendidos en el Hospital san Bartolomé. Los datos obtenidos durante la investigación, por medio de la ficha de recolección de datos, elaborada para los fines de la investigación se ordenaron y procesaron en una computadora personal, valiéndonos del programa SPSS 21.0 para Windows 2007. Resultados: El 52.3 por ciento fueron varones y el 47.7 por ciento mujeres. La media de la edad global fue de 10.5+/-1 años, siendo la mínima de 8 años y la máxima de 12 años. La prevalencia de sobrepeso fue del 35.6 por ciento. La prevalencia de obesidad fue del 10.1 por ciento. En relación a las características epidemiológicas de los pacientes con obesidad y sobrepeso encontramos que la mayoría fueron mujeres, del nivel socioeconómico B, procedentes de zonas urbanas. Los factores de riesgo asociados a sobrepeso fueron el antecedente de obesidad (OR: 3.8), y el consumo de carbohidratos (OR: 4.4). Hubo una diferencia estadísticamente significativa de mayor edad (10.8 versus 10.3), menos horas de actividad física (1 versus 2), y mayor número de ingesta de alimentos en los niños con sobrepeso (3.7 versus 3.2). Hubo una diferencia estadísticamente significativa de mayor número de ingesta de alimentos en los niños con obesidad (3.7 versus 3.3). Conclusión: La prevalencia de sobrepeso y obesidad fue 35.6 por ciento y de 10.1 por ciento respectivamente. Los niños con obesidad y sobrepeso se caracterizaron por ser mujeres, del nivel socioeconómico B, procedentes de zonas urbanas. Los factores de riesgo asociados a sobrepeso y obesidad fueron: el antecedente de obesidad, consumo de carbohidratos, la poca actividad física, y la ingesta de alimentos más de 3 veces al día.
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Masculino , Feminino , Humanos , Criança , Adolescente , Fatores de Risco , Obesidade Infantil , Sobrepeso , Estudos Observacionais como Assunto , Estudos Retrospectivos , Estudos Transversais , Estudos de Casos e ControlesRESUMO
Specific IgE to gliadin was proposed as a marker for wheat dependent exercise induced anaphylaxis, while Tri a 14 was found to induce IgE response in baker's asthma. We evaluated whether these components could be used for discriminating phenotypes of wheat allergy. Twenty-nine patients who were wheat-induced anaphylaxis and/or urticaria (n=21, group I) and baker's asthma (n=8, group II) were enrolled. The prevalence of serum specific IgE to Tri a 14 was higher in group II (25%) than in group I (4.8%), while the serum specific IgE to gliadin was significantly higher in group I (70%) than in group II (12.5%). The cutoff value for predicting the baker's asthma using the ratio of serum specific IgE to Tri a 14 to gliadin was 742.8 optical density×1,000/(kU/L) with high sensitivity and specificity. These findings suggest that Tri a 14/gliadin may be a potential marker for predicting baker's asthma.
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Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Gliadina/imunologia , Imunoglobulina E/sangue , Hipersensibilidade a Trigo/diagnóstico , Hipersensibilidade a Trigo/imunologia , Adulto , Anafilaxia/imunologia , Asma/sangue , Asma/diagnóstico , Asma/imunologia , Biomarcadores/sangue , Feminino , Humanos , Imunoglobulina E/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Fenótipo , Triticum/imunologia , Urticária/imunologiaRESUMO
The study of cross-reactivity in allergy is key to both understanding. the allergic response of many patients and providing them with a rational treatment In the present study, protein microarrays and a co-sensitization graph approach were used in conjunction with an allergen microarray immunoassay. This enabled us to include a wide number of proteins and a large number of patients, and to study sensitization profiles among members of the LTP family. Fourteen LTPs from the most frequent plant food-induced allergies in the geographical area studied were printed into a microarray specifically designed for this research. 212 patients with fruit allergy and 117 food-tolerant pollen allergic subjects were recruited from seven regions of Spain with different pollen profiles, and their sera were tested with allergen microarray. This approach has proven itself to be a good tool to study cross-reactivity between members of LTP family, and could become a useful strategy to analyze other families of allergens.
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Alérgenos/imunologia , Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Proteínas de Plantas/imunologia , Análise Serial de Proteínas/métodos , Alérgenos/química , Reações Cruzadas/imunologia , Eletroforese em Gel de Poliacrilamida , Epitopos/química , Alimentos , Hipersensibilidade Alimentar/imunologia , Geografia , Humanos , Imunoensaio/métodos , Lipídeos/química , Modelos Estatísticos , Pólen , Proteínas Recombinantes/química , Espanha , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Cross-reactivity of plant foods is an important phenomenon in allergy, with geographical variations with respect to the number and prevalence of the allergens involved in this process, whose complexity requires detailed studies. We have addressed the role of thaumatin-like proteins (TLPs) in cross-reactivity between fruit and pollen allergies. A representative panel of 16 purified TLPs was printed onto an allergen microarray. The proteins selected belonged to the sources most frequently associated with peach allergy in representative regions of Spain. Sera from two groups of well characterized patients, one with allergy to Rosaceae fruit (FAG) and another against pollens but tolerant to food-plant allergens (PAG), were obtained from seven geographical areas with different environmental pollen profiles. Cross-reactivity between members of this family was demonstrated by inhibition assays. Only 6 out of 16 purified TLPs showed noticeable allergenic activity in the studied populations. Pru p 2.0201, the peach TLP (41%), chestnut TLP (24%) and plane pollen TLP (22%) proved to be allergens of probable relevance to fruit allergy, being mainly associated with pollen sensitization, and strongly linked to specific geographical areas such as Barcelona, Bilbao, the Canary Islands and Madrid. The patients exhibited >50% positive response to Pru p 2.0201 and to chestnut TLP in these specific areas. Therefore, their recognition patterns were associated with the geographical area, suggesting a role for pollen in the sensitization of these allergens. Finally, the co-sensitizations of patients considering pairs of TLP allergens were analyzed by using the co-sensitization graph associated with an allergen microarray immunoassay. Our data indicate that TLPs are significant allergens in plant food allergy and should be considered when diagnosing and treating pollen-food allergy.
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Reações Cruzadas/imunologia , Hipersensibilidade Alimentar/imunologia , Proteínas de Plantas/imunologia , Análise Serial de Proteínas , Adolescente , Adulto , Criança , Feminino , Alimentos/efeitos adversos , Hipersensibilidade Alimentar/sangue , Frutas/imunologia , Geografia , Humanos , Imunização , Imunoensaio , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Peso Molecular , Proteínas de Plantas/isolamento & purificação , Pólen/imunologia , Espanha , Adulto JovemRESUMO
Hypoallergenic mutants with reduced IgE-binding capacity but which show a similar T-cell response to the corresponding natural allergen are ideal tools for immunotherapy, for preventing a possible anaphylactic shock. An IgE conformational epitope has been identified in Cuc m 2, the major allergen and profilin from melon. Since this epitope is highly conserved in most pollen profilins, it may contribute to an explanation of cross-reactivity between pollen and food profilins. Mutants (Mut 1 and Mut 2) were generated by changing specific residues of the Cuc m 2 epitope to alanine, produced in Escherichia coli, and purified by chromatographic methods. Mut 1 showed a slight reduction in IgE binding but an allergenic activity that was similar to recombinant Cuc m 2, as measured by basophil activation test (BAT) and skin prick test (SPT). By contrast, Mut 2 displayed a substantial reduction in IgE-binding capacity (57%) and positive responses, as determined by BAT (33%) and SPT (50%), when compared to those of rCuc m 2. However, the T-cell proliferation and cytokine production induced by Mut 2 and rCuc m 2 were similar. Thus, this mutant represent potential candidate for immunotherapy of profilin allergies.
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Antígenos de Plantas/imunologia , Cucurbitaceae/imunologia , Imunoglobulina E/imunologia , Profilinas/imunologia , Adolescente , Adulto , Sequência de Aminoácidos , Antígenos de Plantas/química , Células Cultivadas , Sequência Conservada , Cucurbitaceae/química , Epitopos/imunologia , Feminino , Humanos , Imunoterapia , Masculino , Dados de Sequência Molecular , Profilinas/química , Alinhamento de Sequência , Linfócitos T/citologia , Linfócitos T/imunologia , Adulto JovemRESUMO
BACKGROUND: Cross-reactivity among plant food allergens belonging to the nonspecific lipid transfer protein (LTP) family is well known. In contrast, the relationship among these allergens and their putative homologs from olive (Ole e 7) and Parietaria (Par j 1) pollen has not been clarified. METHODS: Sera with specific IgE to LTP allergens were obtained from peach-, mustard- and olive pollen-allergic patients. Purified LTP allergens from foods (peach, apple, mustard and wheat) and pollens (olive, mugwort and Parietaria) were tested by ELISA and ELISA-inhibition assays. RESULTS: Plant food LTP-allergic patients showed a significantly higher number of sera (89-100 vs. 33-64%) with specific IgE and mean specific IgE levels (0.30-1.56 vs. 0.21-0.34 OD units) to the 4 food LTP allergens tested than to olive Ole e 7 and Parietaria Par j 1 pollen. ELISA-inhibition assays indicated cross-inhibition between food LTP allergens but no cross-reactivity between these allergens and Ole e 7 and Par j 1, or, even more, between the LTP allergens from olive and Parietaria pollen. CONCLUSIONS: LTP allergens from olive and Parietaria pollen cross-react neither with allergenic LTPs from plant foods nor between themselves. Therefore, both pollens do not seem to be related with the LTP syndrome.
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Alérgenos/imunologia , Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Reações Cruzadas , Parietaria/imunologia , Proteínas de Plantas/imunologia , Pólen/imunologia , Sequência de Aminoácidos , Hipersensibilidade Alimentar , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Olea/imunologia , Alinhamento de SequênciaRESUMO
The use of peptides from a phage display library selected by binding to a given antibody is a widespread technique to probe epitopes of antigenic proteins. However, the identification of interaction sites mimicked by these peptides on the antigen surface is a difficult task. LocaPep is a computer program developed to localize epitopes using a new clusters algorithm that focuses on protein surface properties. The program is constructed with the aim of providing a flexible computational tool for predicting the location of epitopes on protein structures. As a first set of testing results, the localization of epitope regions in eight different antigenic proteins for which experimental data on their antibody interactions exist is correctly identified by LocaPep. These results represent a disparate sample of biologically different systems. The program is freely available at http://atenea.montes.upm.es.
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Biologia Computacional/métodos , Epitopos/imunologia , Biblioteca de Peptídeos , Proteínas/química , Software , Alérgenos/química , Antígenos de Plantas , Fator VIII/química , Proteína gp120 do Envelope de HIV/química , Humanos , Glicoproteínas de Membrana/química , Modelos Moleculares , Proteínas de Plantas , Conformação Proteica , Estrutura Terciária de Proteína , Prunus , Receptor ErbB-2/química , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave , Glicoproteína da Espícula de Coronavírus , Proteínas do Envelope Viral/químicaRESUMO
BACKGROUND: Profilins are commonly involved in polysensitization of allergic patients; therefore, appropriate markers should be used in component-resolved diagnosis. OBJECTIVE: To evaluate the immunological equivalence between profilins from pollens and plant-derived foods, to be used in component-resolved diagnosis. METHODS: Specific immunoglobulin (Ig) G antibodies against pollen and fruit profilins, as well as sera from patients allergic to mustard, melon, or olive pollen, were used. Purified profilins from mustard seeds, fruit melon, and chenopod and birch pollen were assayed in immunoblotting, enzyme-linked immunosorbent assay (ELISA), and ELISA inhibition assays. RESULTS: Significant correlation was found in the response of purified profilins by ELISA and immunoblotting for both specific IgG and IgE. The highest levels of IgE binding were obtained for olive pollen-allergic patients, which could be related to the route of sensitization. The responses of individual patients to profilins were also similar and independent of the sensitizing source. The inhibition between pairs of allergens was generally higher than 70%, indicating that profilins share most of the IgE epitopes. Modeling of mimotopes in the conformational structure of the implicated profilins supports their strong cross-reactivity obtained experimentally. CONCLUSIONS: No correlation exists between the level of IgE response of individual patients to specific profilins and the corresponding theoretical sensitizing source, suggesting that the sensitization could be attributable to any profilin present in the environment of the patients. This would bear out the use of most profilins as a common marker for polysensitization in component-resolved diagnosis and for therapeutic approaches.
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Alérgenos/imunologia , Reações Antígeno-Anticorpo/imunologia , Imunoglobulina E/imunologia , Plantas Comestíveis/imunologia , Pólen/imunologia , Profilinas/imunologia , Alérgenos/química , Alérgenos/genética , Sequência de Aminoácidos , Animais , Antígenos de Plantas/química , Antígenos de Plantas/genética , Antígenos de Plantas/imunologia , Ligação Competitiva/imunologia , Chenopodium/química , Chenopodium/imunologia , Reações Cruzadas/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/química , Epitopos de Linfócito B/imunologia , Humanos , Hipersensibilidade/diagnóstico , Hipersensibilidade/imunologia , Soros Imunes/imunologia , Immunoblotting , Imunoglobulina G/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Dados de Sequência Molecular , Mostardeira/química , Mostardeira/imunologia , Plantas Comestíveis/química , Pólen/química , Profilinas/química , Profilinas/genética , Coelhos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Homologia de Sequência de AminoácidosRESUMO
Banana fruit allergy is well known, but neither immunoglobulin E recognition patterns to purified plant food allergens nor true prevalences of putative banana allergens have been established. This study aimed to characterize ß-1,3-glucanase and thaumatin-like protein (TLP) as banana allergens, testing them, together with other plant food allergens, in 51 children with allergic reactions after banana ingestion and both positive specific IgE and skin prick test (SPT) to banana. Banana ß-1,3-glucanase and TLP were isolated and characterized. Both banana allergens, together with kiwifruit TLP Act d 2, avocado class I chitinase Pers a 1, palm pollen profilin Pho d 2 and peach fruit lipid transfer protein (LTP) Pru p 3, were tested by in vitro and in vivo assays. Banana ß-1,3-glucanase (Mus a 5) was glycosylated, whereas banana TLP (Mus a 4) was not, in contrast with its homologous kiwi allergen Act d 2. Specific IgE to both banana allergens, as well as to peach Pru p 3, was found in over 70% of sera from banana-allergic children, and Mus a 4 and Pru p 3 provoked positive SPT responses in 6 of the 12 tested patients, whereas Mus a 5 in only one of them. Both peptidic epitopes and cross-reactive carbohydrate determinants were involved in the IgE-binding to Mus a 5, whereas cross-reactivity between Mus a 4 and Act d 2 was only based on common IgE protein epitopes. Profilin Pho d 2 elicited a relevant proportion of positive responses on in vitro (41%) and in vivo (58%) tests. Therefore, Mus a 4 and LTP behave as major banana allergens in the study population, and profilin seems to be also a relevant allergen. Mus a 5 is an equivocal allergenic protein, showing high IgE-binding to its attached complex glycan, and low in vivo potency.
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Alérgenos , Proteínas de Transporte , Hipersensibilidade Alimentar/diagnóstico , Glucana 1,3-beta-Glucosidase , Musa/imunologia , Proteínas de Plantas , Adolescente , Alérgenos/efeitos adversos , Alérgenos/isolamento & purificação , Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Proteínas de Transporte/isolamento & purificação , Criança , Pré-Escolar , Quitinases/imunologia , Quitinases/isolamento & purificação , Reações Cruzadas/imunologia , Feminino , Hipersensibilidade Alimentar/imunologia , Glucana 1,3-beta-Glucosidase/imunologia , Glucana 1,3-beta-Glucosidase/isolamento & purificação , Humanos , Imunoglobulina E/sangue , Lactente , Masculino , Proteínas de Plantas/imunologia , Proteínas de Plantas/isolamento & purificação , Profilinas/imunologia , Profilinas/isolamento & purificação , Prunus/imunologia , Testes CutâneosRESUMO
BACKGROUND: A considerable number of pollen-allergic patients develops allergy to plant foods, which has been attributed to cross-reactivity between food and pollen allergens. The aim of this study was to analyze the differences among pollen-allergic patients with and without plant food allergy. METHODS: Eight hundred and six patients were recruited from 8 different hospitals. Each clinical research group included 100 patients (50 plant food-allergic patients and 50 pollen-allergic patients). Diagnosis of pollen allergy was based on typical case history of pollen allergy and positive skin prick tests. Diagnosis of plant-food allergy was based on clear history of plant-food allergy, skin prick tests and/or plant-food challenge tests. A panel of 28 purified allergens from pollens and/or plant foods was used to quantify specific IgE (ADVIA-Centaur® platform). RESULTS: Six hundred and sixty eight patients (83%) of the 806 evaluated had pollen allergy: 396 patients with pollen allergy alone and 272 patients with associated food and pollen allergies. A comparison of both groups showed a statistically significant increase in the food and pollen allergy subgroup in frequency of: (1) asthma (47 vs. 59%; p < 0.001); (2) positive skin test results to several pollens: Plantago, Platanus, Artemisia, Betula, Parietaria and Salsola (p < 0.001); (3) sensitization to purified allergens: Pru p 3, profilin, Pla a 1 - Pla a 2, Sal k 1, PR-10 proteins and Len c 1. CONCLUSION: Results showed relevant and significant differences between both groups of pollen-allergic patients depending on whether or not they suffered from plant-derived food allergy.
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Hipersensibilidade Alimentar/imunologia , Plantas Comestíveis/imunologia , Rinite Alérgica Sazonal/imunologia , Adulto , Feminino , Humanos , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Testes CutâneosAssuntos
Antígenos de Plantas/análise , Citrullus/efeitos adversos , Cucumis melo/efeitos adversos , Hipersensibilidade Alimentar/imunologia , Proteínas de Plantas/análise , Adulto , Especificidade de Anticorpos , Antígenos de Plantas/imunologia , Feminino , Hipersensibilidade Alimentar/diagnóstico , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Malato Desidrogenase/análise , Malato Desidrogenase/imunologia , Lectinas de Plantas/análise , Lectinas de Plantas/imunologia , Proteínas de Plantas/imunologia , Profilinas/análise , Profilinas/imunologia , Serina Endopeptidases/imunologia , Testes CutâneosRESUMO
Japanese cedar (Cryptomeria japonica) pollen is a major cause of seasonal rhinitis and conjunctivitis in Japan, and an understanding of its full allergen repertoire is prerequisite for the development of future molecular diagnostics and immunotherapeutic strategies. Here we report the identification of a new C. japonica pollen IgE-binding antigen (CJP-8) homologous to lipid transfer proteins (LTPs), a class of plant cross-reactive allergens found in foods, latex, and pollen grains. The cjp-8 cDNA encodes a 165-amino acid polypeptide possessing the conserved eight cysteines characteristic of plant LTP family members. Escherichia coli-expressed recombinant CJP-8 (r-CJP-8) reacted with IgE antibody from Japanese cedar pollinosis patients at a 37.5% frequency (6/16).
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Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Cryptomeria/imunologia , Imunoglobulina E/imunologia , Proteínas de Plantas/imunologia , Pólen/imunologia , Sequência de Aminoácidos , Antígenos de Plantas/genética , Clonagem Molecular , Cryptomeria/genética , Cisteína/imunologia , Humanos , Dados de Sequência Molecular , Proteínas de Plantas/genética , Rinite Alérgica Sazonal/imunologia , Homologia de Sequência de AminoácidosRESUMO
BACKGROUND: Food allergy to wheat-derived foodstuffs is on the rise. Tri a 14, a wheat flour lipid transfer protein (LTP) allergen, has been described as a major allergen associated with baker's asthma and wheat food allergy. Cross-reactivity among LTP allergens leads to the so-called 'LTP syndrome'. METHODS: Eight adult patients showing anaphylaxis after ingestion of wheat-derived foodstuffs were selected. A homemade wheat extract, purified natural (n) and recombinant (r) Tri a 14, and peach fruit and Artemisia pollen LTP allergens Pru p 3 and Art v 3 were subjected to skin prick test, specific IgE determination (ELISA) and IgE immunodetection assays. RESULTS: All tests were positive in the 8 selected patients with the homemade extract. Positive skin prick test responses to nTri a 14, Pru p 3 and Art v 3 were found in 5/8, 6/8 and 4/4 patients, respectively. Specific IgE determined by ELISA assays was detected in 6 to nTri a 14 and rTri a 14, in 4 to Pru p 3 and in 3 to Art v 3 out of 8 individual sera tested, whereas all these sera showed IgE binding to nTri a 14 and Pru p 3 in immunodetection after SDS-PAGE separation. CONCLUSIONS: Tri a 14 seems to be a relevant allergen in patients with anaphylaxis after ingestion of wheat flour foodstuffs, according to in vitro and in vivo results. Clinical history of the analyzed patients, together with sensitization to peach Pru p 3 and Artemisia pollen Art v 3, suggests that 6 of them suffer from LTP syndrome.
Assuntos
Anafilaxia/imunologia , Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Proteínas de Plantas/imunologia , Hipersensibilidade a Trigo/imunologia , Adulto , Alérgenos/imunologia , Antígenos de Plantas/biossíntese , Antígenos de Plantas/genética , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Feminino , Hipersensibilidade Alimentar/imunologia , Gliadina/imunologia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/imunologia , Proteínas Recombinantes/imunologia , Testes Cutâneos , Síndrome , Triticum/química , Triticum/imunologia , Adulto JovemRESUMO
Peach non-specific lipid transfer protein (Pru p 3; nsLTP) has been characterized as the major food allergen in the adult Mediterranean population. Its wheat homologous protein, Tri a 14 has a relevant inhalant allergen in occupational baker's asthma. Different sensitization patterns to these allergens have been found in patients with this latter disorder. The objective of the present study was to characterize IgE epitopes of Tri a 14 and to compare them with those of Pru p 3 using three complementary strategies: the analysis of IgE-binding capacity of decapeptides bound to membrane, the identification of mimotopes using a phage display random peptide library, and the analysis of the surface electrostatic potential of both allergens. Thus, synthetic overlapping decapeptides, covering the Pru p 3 and Tri a 14 amino acid sequences, were used to identify sequential regions involved in recognition of IgE from baker's asthma patients sensitized to both nsLTPs. A phage display library was screened with total IgE from the same patients, and positive clones sequentially selected using the purified allergens, allowed to identify mimotopes (conformational epitopes) of Tri a 14 and Pru p 3. Both sequential regions and mimotopes were localized in the corresponding 3D molecular surface and their electrostatic properties were analyzed. Common sequential regions with strong IgE-binding capacity (residues 31-40 and 71-80) were identified in Tri a 14 and Pru p 3, whereas regions Tri a 14(51-60) and Pru p 3(11-20) were found specific of each allergen. A major conformational epitope (mimotope), L34H35N36R39S40S42D43G74V75L77P78Y79T80, which comprised the two common sequential epitopes, was located in Tri a 14, and a very similar one in Pru p 3. However, differences were detected on the surface electrostatic potential of both mimotopes: a first part (around residues 31-45) showed similar positive features in both allergens, whereas a second part (around residues 74-80) was markedly negative in Tri a 14 but neutral-positive in Pru p 3. Tri a 14 and Pru p 3 have a similar conformational region involved in IgE-binding, although their electrostatic features are different. Additionally, common and specific sequential IgE-binding regions were mapped in both allergens. These findings could be instrumental in understanding the cross-reactivity and specificity of sensitization to both homologous allergens.