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1.
Nutrients ; 13(7)2021 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-34371885

RESUMO

Type 1 diabetes mellitus (T1DM) patients occasionally develop disordered eating behaviors, leading to insulin manipulation without medical consultation, targeting to achieve weight control. In clinical practice, the Diabetes Eating Problem Survey-Revised Version (DEPS-R) questionnaire has been used to evaluate eating disorders in T1DM patients. This study was conducted to validate the factor structure of the Greek version of DEPS-R using Confirmatory Factor Analysis (CFA), to investigate its reliability and convergent validity in Greek T1DM adults and to compare a single factor DEPS-R model with multiple factor models. Participants were 103 T1DM adults receiving insulin, who responded to DEPS-R. Their anthropometric, biochemical and clinical history data were evaluated. The sample presented good glycemic control and 30.1% scored above the established DEPS-R cut-off score for disturbed eating behavior. CFA results revealed that the data fit well to the factor models. The DEPS-R scale had good reliability and was positively linked to BMI, HbA1c, total daily dose and time in range. Model comparison supported the superiority of the 1-factor model, implying that Greek clinicians and practitioners might not have to consider individualized treatment based on various scores across different subscales but they can adopt a single DEPS-R score for an easy and efficient screening for disordered eating.


Assuntos
Diabetes Mellitus Tipo 1/psicologia , Comportamento Alimentar/psicologia , Transtornos da Alimentação e da Ingestão de Alimentos/diagnóstico , Programas de Rastreamento/normas , Inquéritos e Questionários/normas , Adulto , Antropometria , Análise Fatorial , Transtornos da Alimentação e da Ingestão de Alimentos/etiologia , Feminino , Grécia , Humanos , Masculino , Pessoa de Meia-Idade , Psicometria , Reprodutibilidade dos Testes , Traduções
2.
Int Arch Allergy Immunol ; : 1-8, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-34192693

RESUMO

BACKGROUND: Food allergy (FA) in children impacts their own and their family quality of life (QoL). The association of specific FA factors with the various domains of health-related QoL (HRQL) in children is unclear. OBJECTIVE: The aim of this study was to evaluate FA characteristics in primary school children as determinants of components of HRQL. METHODS: Children with FA were recruited from the allergy clinic of a tertiary children's hospital. Demographic and clinical data were retrieved from their records, and 3 HRQL questionnaires were administered: the FA QoL Questionnaire-Child Form (FAQLQ-CF), the FA independent measure (FAIM), and the Pediatric QoL Questionnaire (PedsQL™). Stepwise multiple linear regression analysis was carried out to investigate the correlation between FA characteristics and the scores on the HRQL scales. Bonferroni correction for multiple comparisons was set at p < 0.0002. RESULTS: Of 172 primary schoolchildren with FA invited to take part, 110 participated (response rate 64%), of whom 83 (75.5%) were male, aged 7.5-12.3 years (mean 10.0 ± 1.4) years. From 38 demographic and clinical characteristics, 10 were excluded on initial data analysis and 28 proceeded to bivariate analysis with the scores on FAQLQ-CF, FAIM, PedsQL™, and their subscales. Most of the 28 showed no correlation with the scores; only 4 were entered into multivariate analysis with FAQLQ-CF and PedsQL™ scores, none of which, finally showed significant association. CONCLUSION: The HRQL of children with FA did not depend on gender, age, number, and type of allergen or the characteristics of the most severe allergic reaction.

3.
Allergol Immunopathol (Madr) ; 49(3): 146-152, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33938200

RESUMO

INTRODUCTION AND OBJECTIVES: In order to investigate food allergy's prevalence, risk factors and eating behavior of children with relevant anamnesis, a study was performed in Cypriot primary schools. PATIENTS: A specially composed questionnaire for self-reported adverse reactions to food, created in the context of the EuroPrevall study, was distributed in 13 representative primary schools across the country. Participants were sub-grouped into three groups; healthy (H), those with unconfirmed food hypersensitivity reactions (FA-) and children with a confirmed diagnosis by a physician IgE-mediated food allergy (FA+). Food habits, family health history and lifestyle factors were assessed and groups' outcomes were compared with each other. RESULTS: For the study, 202 questionnaires were completed and returned; 31 children (19 FA- and 12 FA+) reported an adverse food reaction. Significant risk factors for developing FA+ were being the first born or having siblings with asthma, attended a day nursery, but also maternal alcohol drinking during pregnancy, parental smoking and parental occupation in food processing or use of latex gloves. The presence of children in the kitchen during cooking showed a protective role. Dietary habits of FA+ children were significantly diminished in terms of variety and frequency of consumption in comparison to the rest, in which had a greater overlap. CONCLUSION: Further research is required for the interesting risk or protective factors revealing from the current investigation. The negative effect of food allergy in the dietary habits of food allergic children documented in the literature, is strongly supported herein.

4.
Front Immunol ; 12: 661806, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33897707

RESUMO

Breastfeeding not only provides the optimum source of nutrients for the neonate and its first strong shield against infection but also lays the foundation for somatic and psychological bonding between the mother and child. During the current COVID-19 pandemic, although the guidelines of the relevant international and national agencies recommend breastfeeding by SARS-CoV-2-infected mothers, considerable insecurity persists in daily clinical practice regarding the safety of the infants and the perceived advantages and disadvantages of discontinuation of breastfeeding. This is a systematic review of the currently available information regarding the transmissibility of SARS-CoV-2 through or while breastfeeding and the protection against infection that breast milk might provide. The accumulated body of knowledge regarding the role of breast milk in the development of the neonatal immune system and protection against infection by other respiratory viruses is discussed, with a focus on the anti-inflammatory role of the antibodies, microbes, and viruses provided to the infant in breast milk and its relevance to the case of SARS-CoV-2.


Assuntos
Anticorpos Antivirais/imunologia , Aleitamento Materno , COVID-19/imunologia , Transmissão Vertical de Doenças Infecciosas , Leite Humano/imunologia , Complicações Infecciosas na Gravidez/imunologia , SARS-CoV-2/imunologia , COVID-19/patologia , COVID-19/transmissão , Feminino , Humanos , Recém-Nascido , Gravidez , Complicações Infecciosas na Gravidez/patologia
5.
Front Immunol ; 12: 648546, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33763085

RESUMO

Multisystem Inflammatory Syndrome in children (MIS-C) is a rare but devastating complication of coronavirus disease 19 (COVID-19). The development of prognostic biomarkers and more importantly the implementation of new treatment modalities would have a significant impact in clinical practice regarding the outcome of MIS-C. Vitamin D could be a potential candidate. In this mini review we analyze the immunomodulatory role of vitamin D in viral infections and specifically in COVID-19. We also examine the current literature regarding the association of vitamin D with MIS-C and Kawasaki disease. The vitamin D was evaluated not only as a biomarker but also as a nutritional supplement. We concluded that vitamin D levels could be valuable in predicting severe forms of MIS-C and correction of abnormal levels in severe MIS-C may influences its evolution. 25-hydroxyvitamin D3 [25(OH)D3] supplementation raising serum [25(OH)D] concentrations potentially have a favorable effect in reducing the severity of MIS-C in certain circumstances. Further studies are needed to confirm these results.


Assuntos
COVID-19/complicações , Síndrome de Resposta Inflamatória Sistêmica/tratamento farmacológico , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Vitamina D/administração & dosagem , Animais , COVID-19/sangue , COVID-19/tratamento farmacológico , COVID-19/etiologia , COVID-19/virologia , Criança , Humanos , SARS-CoV-2/fisiologia , Síndrome de Resposta Inflamatória Sistêmica/sangue , Vitamina D/sangue
6.
J Allergy Clin Immunol Pract ; 9(1): 225-235.e10, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32916320

RESUMO

BACKGROUND: Walnut allergy is common across the globe, but data on the involvement of individual walnut components are scarce. OBJECTIVES: To identify geographical differences in walnut component sensitization across Europe, explore cosensitization and cross-reactivity, and assess associations of clinical and serological determinants with severity of walnut allergy. METHODS: As part of the EuroPrevall outpatient surveys in 12 European cities, standardized clinical evaluation was conducted in 531 individuals reporting symptoms to walnut, with sensitization to all known walnut components assessed in 202 subjects. Multivariable Lasso regression was applied to investigate predictors for walnut allergy severity. RESULTS: Birch-pollen-related walnut sensitization (Jug r 5) dominated in Northern and Central Europe and lipid transfer protein sensitization (Jug r 3) in Southern Europe. Profilin sensitization (Jug r 7) was prominent throughout Europe. Sensitization to storage proteins (Jug r 1, 2, 4, and 6) was detected in up to 10% of subjects. The walnut components that showed strong correlations with pollen and other foods differed between centers. The combination of determinants best predicting walnut allergy severity were symptoms upon skin contact with walnut, atopic dermatitis (ever), family history of atopic disease, mugwort pollen allergy, sensitization to cat or dog, positive skin prick test result to walnut, and IgE to Jug r 1, 5, 7, or carbohydrate determinants (area under the curve = 0.81; 95% CI, 0.73-0.89). CONCLUSIONS: Walnut-allergic subjects across Europe show clear geographical differences in walnut component sensitization and cosensitization patterns. A predictive model combining results from component-based serology testing with results from extract-based testing and information on clinical background allows for good discrimination between mild to moderate and severe walnut allergy.


Assuntos
Hipersensibilidade Alimentar , Juglans , Nozes , Alérgenos , Animais , Antígenos de Plantas , Gatos , Reações Cruzadas , Cães , Europa (Continente)/epidemiologia , Humanos , Imunoglobulina E
7.
Nutrients ; 12(11)2020 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-33167542

RESUMO

Asthma is a complex chronic inflammatory disorder. Diet's impact on asthma symptoms is controversial. The objective of this pilot crossover, randomized, two-period study was to examine the effect of dietary histamine intake on asthma symptoms in twenty-one children with mild intermittent asthma. Children were randomly assigned to either a high- or low-histamine diet, based on the Mediterranean pattern, for 4 weeks. After a 2-week washout period, patients crossed to the alternative diet for 4 additional weeks. Asthma symptoms were assessed at baseline and after the completion of each diet period by a clinician. Daily symptoms and peak flow were recorded throughout the intervention. Adherence to the dietary intervention was assessed via analysis of four random 24-h recalls, for each intervention period. Eighteen children completed the study. Significantly higher mean air flow obstruction was recorded and a trend for prolonged and more severe symptoms was observed during the high-histamine period. Diet may have an active and direct impact on asthma symptoms. Food choice is affected and/or may affect symptoms in children with mild asthma. Diet intervention is promising yet challenging, for asthma control.


Assuntos
Asma/patologia , Alimentos , Histamina/efeitos adversos , Criança , Dieta , Comportamento Alimentar , Feminino , Humanos , Masculino , Micronutrientes/análise , Projetos Piloto
8.
Food Chem Toxicol ; 127: 61-69, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30826409

RESUMO

Many food innovations rely on the introduction and use of new or modified proteins. New or modified food proteins may lead to major health risks due to their inherent potential to cause food allergy. Currently, the pre-market allergenicity assessment for new or modified food proteins and protein sources relies on methods for identifying allergenic hazards based on characteristics of known allergens. However, there is no general consensus on the allergenicity parameters to use and the criteria that should apply for the evaluation and decisions to be made. In this paper, we propose that the strategy for allergenicity risk assessment of new or modified food proteins and the methodologies applied should be governed by the risk management questions to be answered, reflected in the information needed by risk managers to enable their informed decision making. We generated an inventory of health outcome-related assessment parameters and criteria potentially important for risk management decision-making and we discuss the implications of selecting different optional criteria (e.g. cut-off values) for what could be accepted as safe with regards to the health outcomes in the (at risk) population. The impact of these various options on both method development and risk management practices was investigated.


Assuntos
Alérgenos/imunologia , Proteínas na Dieta/imunologia , Hipersensibilidade Alimentar/prevenção & controle , Imunoglobulina E/metabolismo , Alérgenos/efeitos adversos , Tomada de Decisões , Proteínas na Dieta/efeitos adversos , Hipersensibilidade Alimentar/imunologia , Humanos , Medição de Risco , Gestão de Riscos
10.
Mol Biotechnol ; 56(6): 498-506, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24198217

RESUMO

Allergy to banana fruit appears to have become an important cause of fruit allergy in Europe. Among five allergens that have been found, beta-1,3-glucanase denoted as Mus a 5 was identified as a candidate allergen for the component-resolved allergy diagnosis of banana allergy. Because of the variations in protein levels in banana fruit, in this study Mus a 5 was produced as a fusion protein with glutathione-S-transferase in Escherichia coli. The recombinant Mus a 5 was purified under native conditions by a combination of affinity, ion-exchange, and reversed phase chromatography. N-terminal sequence was confirmed by Edman degradation and 55 % of the primary structure was identified by mass fingerprint, while the secondary structure was assessed by circular dichroism spectroscopy. IgG reactivity of recombinant protein was shown in 2-D immunoblot with anti-Mus a 5 antibodies, while IgG and IgE binding to natural Mus a 5 was inhibited with the recombinant Mus a 5 in immunoblot inhibition test. IgE reactivity of recombinant Mus a 5 was shown in ELISA within a group of ten persons sensitized to banana fruit. Recombinant Mus a 5 is a novel reagent suitable for the component-resolved allergy diagnosis of banana allergy.


Assuntos
Alérgenos/genética , Hipersensibilidade Alimentar/genética , Glucana 1,3-beta-Glucosidase/genética , Alérgenos/imunologia , Sequência de Aminoácidos , Animais , Europa (Continente) , Frutas/imunologia , Glucana 1,3-beta-Glucosidase/imunologia , Humanos , Camundongos , Musa , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia
11.
Mol Nutr Food Res ; 56(3): 446-53, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22162266

RESUMO

SCOPE: Banana fruit has become an important cause of fruit allergy in the recent years. Among the five registered IUIS allergens, Mus a 1 and Mus a 2 have been characterized in detail. In this study, molecular characterization and evaluation of the allergenic properties of ß-1,3-glucanase from banana (Musa acuminata), denoted as Mus a 5, were performed. METHODS AND RESULTS: The gene of Mus a 5 was cloned and sequenced. The obtained cDNA revealed a novel Mus a 5 isoform with an open reading frame encoding a protein of 340 amino acids comprising a putative signal peptide of 28 amino acid residues. By MALDI-TOF analysis Mus a 5 isolated from banana fruit revealed a molecular mass of 33451±67 Da. Two Mus a 5 isoforms (pI 7.7 and 8.0) were detected by 2D immunoblot with an identical N-terminal sequence. By mass fingerprint, 76 and 83% of the primary structure was confirmed for the two mature Mus a 5 isoforms, respectively. IgE reactivity to Mus a 5 was found in 74% of patients sensitized to banana fruit. Upregulation of basophil activation markers CD63 and CD203c was achieved with Mus a 5 in a concentration-dependent manner. CONCLUSION: Mus a 5 is a functional allergen and a candidate for the component-resolved allergy diagnosis of banana allergy.


Assuntos
Alérgenos/imunologia , Alérgenos/isolamento & purificação , Antígenos de Plantas/imunologia , Antígenos de Plantas/isolamento & purificação , Musa/química , Proteínas de Plantas/isolamento & purificação , Adolescente , Adulto , Sequência de Aminoácidos , Antígenos de Plantas/química , Clonagem Molecular , Endo-1,3(4)-beta-Glucanase/imunologia , Feminino , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Frutas/química , Humanos , Imunoglobulina E/metabolismo , Masculino , Dados de Sequência Molecular , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Isoformas de Proteínas/química , Pirofosfatases/genética , Pirofosfatases/metabolismo , Testes Cutâneos/métodos , Tetraspanina 30/genética , Tetraspanina 30/metabolismo , Regulação para Cima , Adulto Jovem
12.
Clin Transl Allergy ; 1(1): 10, 2011 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-22409883

RESUMO

BACKGROUND: European legislators and wine producers still debate on the requirement for labeling of wines fined with potentially allergenic food proteins (casein, egg white or fish-derived isinglass). We investigated whether wines fined with known concentrations of these proteins have the potential to provoke clinical allergic reactions in relevant patients. METHODS: In-house wines were produced for the study, fined with different concentrations of casein (n = 7), egg albumin (n = 1) and isinglass (n = 3). ELISA and PCR kits specific for the respective proteins were used to identify the fining agents. Skin prick tests and basophil activation tests were performed in patients with confirmed IgE-mediated relevant food allergies (n = 24). A wine consumption questionnaire and detailed history on possible reactions to wine was obtained in a multinational cohort of milk, egg or fish allergic patients (n = 53) and patients allergic to irrelevant foods as controls (n = 13). RESULTS: Fining agents were not detectable in wines with the available laboratory methods. Nevertheless, positive skin prick test reactions and basophil activation to the relevant wines were observed in the majority of patients with allergy to milk, egg or fish, correlating with the concentration of the fining agent. Among patients consuming wine, reported reactions were few and mild and similar with the ones reported from the control group. CONCLUSION: Casein, isinglass or egg, remaining in traces in wine after fining, present a very low risk for the respective food allergic consumers. Physician and patient awareness campaigns may be more suitable than generalized labeling to address this issue, as the latter may have negative impact on both non-allergic and allergic consumers.

13.
Pediatr Allergy Immunol ; 21(1 Pt 1): 90-5, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19744220

RESUMO

The rise of asthma prevalence in children observed the last years might be related to several dietary factors/components as suggested by several researchers. We aimed to evaluate the potential relationship between certain nutrients intake and asthma occurrence in a population of pre-school children. In the framework of the cross-sectional study 'Growth, Exercise and Nutrition Epidemiological Study In pre-schoolers', data were collected from 1964 children, aged 24-72 months, living in five different counties in Greece. The International Study of Asthma and Allergies in Childhood questionnaire was used to assess asthma related outcomes. Dietary intake was assessed with 3 days diet records. The prevalence of ever wheeze, current wheeze and diagnosed asthma was 37.7%, 27.5% and 10.5% respectively. Dietary intake of magnesium had a 0.5% and 0.6% increase in the reported risk of current wheeze and diagnosed asthma respectively. On the contrary a decrease in the prevalence of ever (OR: 0.997, 95% CI: 0.995-1.000) and current wheeze (OR: 0.996, 95% CI: 0.993-0.999) was associated with vitamin C intake. Calcium intake slightly decreased the risk of current wheeze (OR: 0.999, 95% CI: 0.998-0.999). An increase of 2% of the risk of reporting ever or current wheeze was associated with mono-unsaturated fatty acid intake. Magnesium intake was the only independent predictor for doctor's diagnosed asthma. We conclude that dietary intake of vitamin C and calcium seem to have a protective effect on the incidence of wheeze in pre-school children, whereas magnesium and mono-unsaturated fatty acid may have a harmful role.


Assuntos
Asma , Dieta , Ingestão de Energia , Sons Respiratórios , Ácido Ascórbico , Asma/diagnóstico , Asma/epidemiologia , Índice de Massa Corporal , Cálcio , Criança , Pré-Escolar , Estudos Transversais , Ácidos Graxos Insaturados , Feminino , Grécia/epidemiologia , Inquéritos Epidemiológicos , Humanos , Magnésio , Masculino , Prevalência , Sons Respiratórios/diagnóstico , Inquéritos e Questionários
15.
Mol Nutr Food Res ; 52 Suppl 2: S196-207, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18504705

RESUMO

Allergic reaction following fish consumption can trigger life-threatening reactions in predisposed individuals. Parvalbumins from different species have been identified as the major fish allergens. There are two distinct phylogenetic lineages of parvalbumins, alpha and beta. Most allergic reactions are caused by beta-parvalbumins. We cloned and expressed cDNAs encoding cod (Gadus morhua) and carp (Cyprinus carpio) beta-parvalbumins and purified natural cod beta-parvalbumin. CD spectra of the purified proteins showed that their overall secondary structure contents were very similar. No differences in thermal stability were monitored in the calcium-bound or calcium-depleted form of natural cod parvalbumin. IgE reactivity was assessed using 26 sera of fish allergic patients from Spain, The Netherlands, and Greece in immunoblot and ELISA experiments. Twenty-five of the 26 patients with IgE reactivity to native and recombinant cod parvalbumin also reacted to the recombinant carp parvalbumin. IgE inhibition assays were performed using cod and carp extracts and purified recombinant parvalbumin of cod and carp. High crossreactivity among cod and carp parvalbumins was observed in immunoblots as well as in fluid phase assays. Natural and recombinant parvalbumins gave comparable results when performing various in vitro diagnostic assays.


Assuntos
Alérgenos/química , Carpas/imunologia , Gadiformes/imunologia , Parvalbuminas/química , Sequência de Aminoácidos , Animais , Cálcio/metabolismo , Dicroísmo Circular , Clonagem Molecular , Reações Cruzadas , Concentração de Íons de Hidrogênio , Imunoglobulina E/metabolismo , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Dados de Sequência Molecular , Parvalbuminas/imunologia , Parvalbuminas/isolamento & purificação , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação
16.
Int Arch Allergy Immunol ; 146(2): 156-61, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18204282

RESUMO

BACKGROUND: Although development of clinical tolerance is the rule in allergy to cow's milk (CM), food challenges are required in order to reintroduce CM into the patient's diet. Less 'invasive' procedures able to predict tolerance would be useful as clinical tools. The purpose of this study was to identify potential risk factors for clinical reactivity in CM-allergic children assessed for CM reintroduction. METHODS: One hundred and sixteen open challenges performed in children 10-47 months old with IgE-mediated allergy to CM, in order to reintroduce CM into the diet, were retrospectively evaluated. Specific IgE (sIgE) levels assessed by the CAP System FEIA and skin prick tests (SPT) were obtained at diagnosis and prechallenge. Demographic parameters and measures of sIgE were evaluated as potential predictors of a positive challenge in univariate and multivariate logistic regression models. RESULTS: Twenty-four out of 116 challenges were positive, 9 of which required the use of adrenaline. In order of performance, prechallenge sIgE <3.94 kU/l, the combination of SPT and sIgE or an SPT wheal <4 mm could correctly predict a negative challenge outcome, whereas values of SPT >7.5 mm or sIgE >25.4 kU/l, or their combination, had a high positive predictive value. The presence of atopic dermatitis did not affect the predictive accuracy of these values. CONCLUSIONS: Milk sIgE level prechallenge is a useful predictor of challenge outcome in patients with milk allergy. SPT do not offer an additional predictive value, but can be used when sIgE is not available.


Assuntos
Hipersensibilidade a Leite/diagnóstico , Hipersensibilidade a Leite/imunologia , Leite/efeitos adversos , Leite/imunologia , Angioedema/imunologia , Animais , Criança , Pré-Escolar , Feminino , Gastroenteropatias/imunologia , Humanos , Imunoglobulina E/sangue , Lactente , Recém-Nascido , Masculino , Hipersensibilidade Respiratória/imunologia , Estudos Retrospectivos , Testes Cutâneos , Urticária/imunologia
17.
Mol Nutr Food Res ; 51(3): 360-6, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17309118

RESUMO

Standardized allergen extracts are needed for diagnosis and therapy purposes. For grapes, standardization is hampered by low protein and high tannin and pectin concentrations. The aim of the current study was to develop an optimized method for the extraction of grape proteins and possibly extend this to other fruits. Several existing or modified extraction methods were compared by means of protein concentration determination, SDS-PAGE, immunoblotting and radioallergosorbent test (RAST). An optimized extraction protocol was obtained in which we combined a high concentration of plant tissue, a concentrated, enriched and neutral buffer able to remove sugars and keep proteins soluble and a bivalent buffer for pectin removal. Both the quantitative (protein concentration) and qualitative parameters (SDS-PAGE protein patterns and IgE reactivity) were compared to standard protocols and commercial extracts used as diagnostic tools in the clinical practice. This method proved to be the most efficient mainly compared to the standard Björksten protocol in extracting the low molecular weight proteins, including the major grape allergen (lipid transfer protein, Vit v 1). It proved to be an easy, low cost and reproducible method proposed to prepare grape extracts that could replace the commercially available ones, used for diagnosis and possibly extend the method to other fruits especially in extracting LTPs.


Assuntos
Alérgenos/isolamento & purificação , Hipersensibilidade Alimentar/diagnóstico , Frutas/imunologia , Vitis/imunologia , Adolescente , Adulto , Criança , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Imunoglobulina E/imunologia , Masculino , Teste de Radioalergoadsorção , Testes Cutâneos
18.
Int Arch Allergy Immunol ; 143(2): 92-102, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17228170

RESUMO

BACKGROUND: Grape allergy is considered rare; grape lipid transfer protein (LTP; Vit v 1), an endochitinase and a thaumatin-like protein (TLP) have been reported as grape allergens. A considerable number of patients have referred to our department for severe reactions to grapes, and several IgE binding proteins were detected. OBJECTIVES: The aim of this study was to identify and characterise the allergens involved in severe allergic reactions to grapes and describe the population in which they occur. METHODS: Patients with reported severe allergic reactions to grapes (n = 37) are described. Grape allergens were purified/fractionated by a combination of chromatographic techniques, identified by proteomic analysis and biochemically characterised. Immunoreactivity was assessed by blot (inhibitions) and RAST (inhibitions), and skin prick tests were performed with the isolated allergens. RESULTS: All subjects were polyallergic, sensitised and reactive to several additional foods and pollen. All patients were sensitised to grape LTP. A 28-kDa expansin, a 37.5-kDa polygalacturonase-inhibiting protein, a 39-kDa beta-1,3-glucanase and a 60-kDa protein were identified as minor grape allergens. Endochitinase and TLP did not play a role. Inhibition experiments revealed the possible cross-reactive role of LTP for clinical sensitivities to other LTP-containing plant foods, but also the involvement of cross-reactive carbohydrate determinants of minor allergens in IgE cross-reactivity. CONCLUSIONS: LTP is the major grape allergen, while additional minor allergens may contribute to clinical reactivity. Severe grape allergy presents in atopic patients who frequently react to other LTP-containing, plant-derived foods. The 'LTP syndrome' is the appropriate term to describe this condition.


Assuntos
Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Imediata/imunologia , Vitis/imunologia , Adolescente , Adulto , Western Blotting , Criança , Pré-Escolar , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Imunoglobulina E/sangue , Masculino , Proteômica , Teste de Radioalergoadsorção , Vitis/química
19.
J Allergy Clin Immunol ; 118(2): 473-80, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16890774

RESUMO

BACKGROUND: Severe grape allergy has been linked to lipid transfer protein (LTP) sensitization. LTPs are known to be resistant to pepsin digestion, although the effect of gastroduodenal digestion on its allergenicity has not been reported. OBJECTIVE: We sought to investigate the effect of gastric and gastroduodenal digestion on the allergenic activity of grape LTP. METHODS: The proteolytic stability of grape LTP was investigated by using an in vitro model of gastrointestinal digestion. The allergenicity of LTP and its digesta was assessed in vitro by means of IgE immunoblotting, RASTs, and in vivo skin prick tests in the same patients with grape allergy. RESULTS: Grape LTP was resistant to gastric digestion, and yielded a 6000-d relative molecular mass C-terminally trimmed fragment after duodenal digestion. This fragment retained the in vitro IgE reactivity of the intact protein. Inclusion of phosphatidylcholine during gastric digestion protected the LTP to a limited extent against digestion. Digestion did not affect the in vivo (skin prick test) biologic activity of LTP. CONCLUSION: The allergenic activity of grape LTP was highly resistant to in vitro digestion. This property might facilitate sensitization through the gastrointestinal tract and might also potentiate the ability of LTPs to elicit severe allergic reactions in sensitized individuals. CLINICAL IMPLICATIONS: Purified natural allergens will facilitate the development of component-resolved diagnostic approaches, including allergen chips. This study contributes to our understanding of the role digestion plays in symptom elicitation in true food allergy.


Assuntos
Proteínas de Transporte/imunologia , Digestão/fisiologia , Duodeno/metabolismo , Suco Gástrico/metabolismo , Vitis/imunologia , Alérgenos/imunologia , Antígenos de Plantas , Basófilos/imunologia , Criança , Pré-Escolar , Hipersensibilidade Alimentar/sangue , Hipersensibilidade Alimentar/imunologia , Liberação de Histamina , Humanos , Imunoglobulina E/sangue , Masculino , Proteínas de Plantas
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