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1.
J Chromatogr A ; 1643: 462076, 2021 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-33789193

RESUMO

The present research is focused on the preliminary evaluation, in particular in relation to the advisable operational conditions, of a novel low duty cycle flow modulator. In such a respect, a fast comprehensive two-dimensional gas chromatography-mass spectrometry method is herein proposed. Applications on a C7-C30 series of alkanes, 64 fragrance allergens (plus 2 internal standards), and 5 perfumes, were carried out by using two different column sets, low-polarity + medium-polarity and low-polarity + low-polarity. In both cases, the first column was of dimensions 10 m × 0.25 mm ID × 0.25 µm df, while the second one was of dimensions 1 m × 0.10 mm ID × 0.10 µm df. A modulation period of 700 ms, with a re-injection period of 80 ms, was used in order to obtain a higher duty cycle (measured to be approx. 0.04). Absolute quantification of the allergens was carried out by using two internal standards, namely 1,4-dibromobenzene and 4,4'-dibromobiphenyl. In terms of limits of quantification the instrumental response was characterized by a wide variability, ranging between 9 ppb and 5.4 ppm for both column sets. A total number of 97 fragrance allergens were identified and quantified in five commercial perfumes.


Assuntos
Alérgenos/análise , Cromatografia Gasosa/métodos , Perfumes/química , Alcanos/análise , Alcanos/química , Alérgenos/química , Limite de Detecção , Perfumes/normas , Padrões de Referência , Espectrometria de Massas por Ionização por Electrospray
2.
Nutrients ; 13(2)2021 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-33578927

RESUMO

Detailed information about the amount of allergenic protein ingested by the patient prior to an allergic reaction yields valuable information for the diagnosis, guidance and management of food allergy. However, the exact amount of ingredients is often not declared on the label. In this study the feasibility was studied for estimating the amount of allergenic protein from milk, eggs, peanuts and hazelnuts in frequently consumed composite and non-composite foods and per bite or sip size in different age groups in the Netherlands. Foods containing milk, egg, peanut or hazelnut most frequently consumed were selected for the age groups 2-3, 4-6 and 19-30 years. If the label did not yield clear information, the amount of allergenic protein was estimated based on food labels. Bite or sip sizes were determined in these age groups in 30 different foods. The amount of allergenic protein could be estimated in 47/70 (67%) of composite foods, which was complex. Estimated protein content of milk, egg, peanut and hazelnut was 2-3 g for most foods but varied greatly from 3 to 8610 mg and may be below threshold levels of the patient. In contrast, a single bite or sip can contain a sufficient amount of allergenic protein to elicit an allergic reaction. Bite and sip sizes increased with age. In every day practice it is hard to obtain detailed and reliable information about the amount of allergenic protein incorporated in composite foods. We encourage companies to disclose the amount of common allergenic foods on their labels.


Assuntos
Alérgenos/análise , Proteínas na Dieta/análise , Hipersensibilidade Alimentar/imunologia , Rotulagem de Alimentos , Adolescente , Adulto , Alérgenos/imunologia , Animais , Arachis/química , Arachis/imunologia , Criança , Pré-Escolar , Corylus/química , Corylus/imunologia , Dieta , Proteínas na Dieta/imunologia , Relação Dose-Resposta Imunológica , Ovos/análise , Feminino , Hipersensibilidade Alimentar/diagnóstico , Humanos , Masculino , Leite/química , Leite/imunologia , Países Baixos , Inquéritos e Questionários , Adulto Jovem
3.
J Chromatogr A ; 1639: 461877, 2021 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-33545578

RESUMO

An analytical approach using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed to simultaneously detect Fagopyrum esculentum Moench (buckwheat) and cereals containing gluten (Triticum species including wheat and spelt, rye, barley, and oats) that were specified in regulations for food allergen labeling on processed foods. Trypsin-digested peptides were purified from different processed food commodities and heptapeptides derived from buckwheat 13S globulin (GFIVQAR, m/z 395.8 [precursor] > 177.0 [product]) and Triticum low molecular weight glutenin (QIPEQSR, m/z 429.3 [precursor] > 616.2 [product]) were specifically detected each species at levels as low as 0.050-0.056 µg/L and 0.028-0.032 µg/L, respectively. Detection of these synthetic peptides was quantitative to over 100 µg/L by reference to the synthetic peptide calibration curves and at recovery rates, 76.6 ± 4.1%-104.8 ± 17.1% and 82.4 ± 2.0%-105.8 ± 5.3%, for GFIVQAR and QIPEQSR, respectively, when 1-1,000 µg of these peptides were spiked into a retort tomato sauce for pasta or dried instant soup. In combination with LC-MS/MS detection methods specific to other cereals containing gluten (rye, barley, and oats), the developed analytical approach was applicable to a wide variety of processed food commodities for food allergen labeling.


Assuntos
Alérgenos/análise , Grão Comestível/química , Análise de Alimentos , Hipersensibilidade Alimentar/diagnóstico , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão , Peptídeos/análise
4.
Sci Total Environ ; 767: 145426, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33550056

RESUMO

Bioaerosols play a major role in the plant life of ecosystems. In addition, they have a profound impact on human health, since they may cause lung diseases or allergies. The key objective of this study is to assess the below cloud scavenging effect of rainfall on pollen concentration. The analysis is based on a sampling carried out in León, Spain, between 2015 and 2018. The rainfall variables and the pollen concentrations have been obtained with a disdrometer and a volumetric Hirst type spore-trap, respectively. In order to evaluate the scavenging, three parameters have been calculated: scavenging efficiency (through the concentration-weighted average (%ΔC)), the scavenging coefficient (λ) and the percentage of events with a decrease in pollen concentration (%ES) also called events with effective scavenging. 71% of rain events presented an effective scavenging that affected all types of pollen. The %ΔC mean value of total pollen was 24 ± 18% (positive values indicate an effective scavenging) and the types of pollen with the highest values were Castanea and Cupressaceae (71 and 40%, respectively). A linear model (R2 = 0.94) to estimate the pollen concentration after rain was built with variables such as pollen concentration before rain and other variables from a weather station and a disdrometer. Furthermore, we have shown the possibility of knowing in real time the probable Cupressaceae pollen concentration, from the initial pollen concentration and the physical parameters of rain (such as raindrop size, rain intensity or volume swept by raindrops in their falling path).


Assuntos
Poluentes Atmosféricos , Poluentes Atmosféricos/análise , Alérgenos/análise , Ecossistema , Humanos , Pólen/química , Estações do Ano , Espanha , Tempo (Meteorologia)
5.
Food Chem ; 347: 129064, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33486358

RESUMO

A one-step mild extraction of total wheat protein fractions was developed in this study, and the allergic cross-reactivity among dietary cereals were assessed by SDS-PAGE, western blotting, indirect ELISA, and inhibition ELISA using sera from 12 wheat allergic patients. The fractions of albumin, globulin, gliadin and glutenins in wheat flour can be obtained by a one-step extraction with Na2CO3-NaHCO3 (20 mM, pH 9.6, 0.5 M NaCl, 40% ethanol, 1 mM PMSF) in comparison to sequential extractions. Results showed high cross-reactivity in wheat, barley and rye due to close resemblance and high sequence identity (>50%), whereas nearly negligible cross-reactivity among rice, buckwheat, and quinoa was observed. Our research findings suggest that people with wheat allergy should rely primarily on the use of rice, quinoa and non-grain buckwheat, which is an effective substitute for wheat, while those with hypersensitivity should avoid the use of barley and rye in their diet.


Assuntos
Alérgenos/análise , Grão Comestível/química , Extração em Fase Sólida/métodos , Triticum/metabolismo , Hipersensibilidade a Trigo/diagnóstico , Adulto , Alérgenos/imunologia , Alérgenos/isolamento & purificação , Sequência de Aminoácidos , Chenopodium quinoa/metabolismo , Reações Cruzadas , Grão Comestível/imunologia , Grão Comestível/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Glutens/análise , Glutens/imunologia , Hordeum/metabolismo , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Proteínas de Plantas/isolamento & purificação , Alinhamento de Sequência , Hipersensibilidade a Trigo/patologia
6.
Methods Mol Biol ; 2241: 139-148, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33486734

RESUMO

Eosinophils are important for tissue homeostasis and host responses to pathogens and allergens. The impact of eosinophils within tissues depends in part on whether cytotoxic proteins in crystalloid granules are released. Determinants of eosinophil motility and loss of granule contents are incompletely understood. The goal of this chapter is to present methods to study the effects of potential mediators on purified human blood eosinophils interacting with adhesive proteins found in extracellular matrix. We show that differential interference contrast video-enhanced microscopy and a bead-clearing assay provide complementary information about how different mediator-adhesive protein combinations direct eosinophil motility and granule fate. The former method is rich in information about cell shape, pattern of movement, and state of granules whereas the latter method lends itself to quantification and interrogation of multiple conditions in replicate.


Assuntos
Ensaios de Migração Celular/métodos , Movimento Celular/imunologia , Eosinófilos/citologia , Alérgenos/análise , Proteínas Sanguíneas/análise , Grânulos Citoplasmáticos/química , Proteínas Granulares de Eosinófilos/química , Matriz Extracelular/imunologia , Citometria de Fluxo/métodos , Humanos , Microscopia/métodos
7.
J Environ Manage ; 282: 111963, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33465718

RESUMO

Pollen grains emitted by urban vegetation are the main primary biological airborne particles (PBAPs) which alter the biological quality of urban air and have a significant impact on human health. This work analyses the interactions which exist between pollen-type PBAPs, meteorological variables, and air pollutants in the urban atmosphere so that the complex relationships and trends in future scenarios of changing environmental conditions can be assessed. For this study, the 1992-2018 pollen data series from the city of Granada (southeast Spain) was used, in which the dynamics of the total pollen as well as the 8 main pollen types (Cupressaceae, Olea, Pinus, Platanus, Poaceae, Populus, Quercus and Urticaceae) were analysed. The trend analysis showed that all except Urticaceae trended upward throughout the series. Spearman's correlations with meteorological variables showed that, in general, the most influential variables on the pollen concentrations were the daily maximum temperature, relative humidity, water vapor pressure, global radiation, and insolation, with different effects on different pollen types. Parallel analysis by neural networks (ANN) confirmed these variables as the predominant ones, especially global radiation. The correlation with atmospheric pollutants revealed that ozone was the pollutant with the highest influence, although some pollen types also showed correlation with NO2, SO2, CO and PM10. The Generalized Linear Models (GLM) between pollen and pollutants also indicated O3 as the most prominent variable. These results highlight the active role that pollen-type PBAPs have on urban air quality by establishing their interactions with meteorological variables and pollutants, thereby providing information on the behaviour of pollen emissions under changing environmental conditions.


Assuntos
Poluentes Atmosféricos , Poluentes Ambientais , Poluentes Atmosféricos/análise , Alérgenos/análise , Cidades , Monitoramento Ambiental , Humanos , Estações do Ano , Espanha
8.
Sci Total Environ ; 757: 143924, 2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33310575

RESUMO

BACKGROUND: Studies have demonstrated an adverse role of outdoor allergens on respiratory symptoms. It is unknown whether this effect is independent or synergistic of outdoor air pollutants. METHODS: We systematically reviewed all epidemiological studies that examined interaction effects between counts of outdoor airborne allergens (pollen, fungal spores) and air pollutants, on any respiratory health outcome in children and adults. We searched the MEDLINE, EMBASE and Scopus databases. Each study was summarized qualitatively and assessed for quality and risk of bias (International Prospective Register for Systematic Reviews, registration number CRD42020162571). RESULTS: Thirty-five studies were identified (15 timeseries, eight case-crossovers, 11 panels and one cohort study), of which 12 reported a significant statistical interaction between an allergen and air pollutant. Eight interactions were related to asthma outcomes, including one on lung function measures and wheeze, three to medical consultations for pollinosis and one to allergic symptoms (nasal, ocular or bronchial). There was no consensus as to which allergen or air pollutant is more likely to interact. No study investigated whether interactions are stronger in atopic individuals. CONCLUSION: Despite strong evidence from small experimental studies in humans, only a third of studies identified significant allergen-pollutant interactions using common epidemiological study designs. Exposure misclassification, failure to examine subgroups at risk, inadequate statistical power or absence of population-level effects are possible explanations.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Adulto , Poluentes Atmosféricos/efeitos adversos , Poluentes Atmosféricos/análise , Poluição do Ar/efeitos adversos , Poluição do Ar/análise , Alérgenos/análise , Criança , Estudos de Coortes , Humanos , Pólen/química
9.
Methods Mol Biol ; 2178: 357-376, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33128761

RESUMO

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is largely recognized as an important tool in the analysis of many biomolecules such as proteins and peptides. The MS analysis of digested peptides to identify a protein or some of its modifications is a key step in proteomics. MALDI-MS is well suited for the peptide mass fingerprinting (PMF) technique, as well as selected fragmentation of various precursors using collisional-induced dissociation (CID) or post-source decay (PSD).In the last few years, MALDI-MS has played a significant role in food chemistry, especially in the detection of food adulterations, characterization of food allergens, and investigation of protein structural modifications induced by various industrial processes that could be an issue in terms of food quality and safety.Here, we present simple extraction protocols of allergenic proteins in food commodities such as milk, egg, hazelnut , and lupin seeds. Classic bottom-up approaches based on Sodium Dodecyl Sulphate (SDS) gel electrophoresis separation followed by in-gel digestion or direct in-solution digestion of whole samples are described. MALDI-MS and MS /MS analyses are discussed along with a comparison of data obtained by using the most widespread matrices for proteomic studies, namely, α-cyano-4-hydroxy-cinnamic acid (CHCA) and α-cyano-4-chloro-cinnamic acid (CClCA). The choice of the most suitable MALDI matrix is fundamental for high-throughput screening of putative food allergens.


Assuntos
Alérgenos/análise , Análise de Alimentos , Hipersensibilidade Alimentar , Proteômica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Alérgenos/química
10.
J Food Sci ; 86(1): 68-77, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33336377

RESUMO

Currently, there has been a surge of interest in revealing the interactions between plasma and food matrices. In this study, we investigated the impacts of atmospheric cold plasma (ACP) treatment on the structural, physicochemical and allergenic characteristics of soybean protein isolate (SPI). SPI dispersions were subjected to ACP treatments at different frequencies (80 to 100 Hz) and durations (1 to 10 min) to investigate the effects of exposing conditions. Results showed that ACP induced reactive oxygen species-mediated oxidation of soy proteins, resulting in modifications in the secondary and ternary structures of SPI. As a consequence, functional properties of SPI, such as emulsifying (56 to 168%, compared with control) and foaming properties (60 to 194%) were influenced by varying degrees. In addition, under certain circumstance (120 Hz, 5 min), the IgE-binding level of SPI was decreased by up to 75%, when compared to the control. Moderate treatment yielded products with improved functionality and reduced allergenicity, while extensive exposure induced a loss of vendibility due to protein aggregation. PRACTICAL APPLICATION: In this study, we demonstrated for the first time, that plasma species reacted with soybean proteins, resulting in spatial structural changes which are closely related with protein functionality and allergenicity. ACP interacts with macromolecules in aqueous systems and thus can be an alternative and promising nonthermal approach in modifying soybean proteins, whereas the exact role of different processing parameters needs to be well-elaborated.


Assuntos
Alérgenos/análise , Manipulação de Alimentos/métodos , Proteínas de Soja/química , Proteínas de Soja/imunologia , Fenômenos Químicos , Emulsões/química , Oxirredução , Gases em Plasma , Conformação Proteica , Espécies Reativas de Oxigênio/química , Água/química
11.
Allergol. immunopatol ; 48(6): 576-581, nov.-dic. 2020. graf, tab
Artigo em Inglês | IBECS | ID: ibc-199245

RESUMO

INTRODUCTION: Eosinophilic esophagitis (EoE) is a chronic, local immune-mediated esophageal disease that has been on the increase lately. There is currently enough evidence to conclude that EoE is an allergic disorder triggered by food allergens, with cow's milk (CM) being the most frequent. Dietary intervention is the first-line approach. This study aimed to assess the clinical characteristics, the diagnostic method, and the prognosis of patients whose culprit food was CM, as opposed to other triggers. METHODS: Children with EoE evaluated in our pediatric Allergy Department were retrospectively studied from 2004 to 2017. We collected clinical variables, diagnostic protocol, treatment, and follow-up data. We compared patients whose culprit food was CM and patients with EoE due to other causative agents. RESULTS: We analyzed 31 children with EoE and found the causative food to be cow's milk in 14 (45%). Clinical characteristics were similar in patients with EoE due to milk or any other cause. Eight of 14 patients with milk-induced EoE (57.14%) presented positive skin prick test results against cow's milk. All patients had positive IgE against cow's milk. None of the patients had any other food as the trigger. The median follow-up was 2.68 years (6 months to 9 years) with initial remission of 100%. CONCLUSION: Testing-based elimination diets effectively treated all of the patients with milk-induced EoE. The advantage of this diagnostic protocol is that it required a mean of only two foods to be tested, significantly smaller number than in empiric diets


No disponible


Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Esofagite Eosinofílica/diagnóstico , Esofagite Eosinofílica/imunologia , Hipersensibilidade a Leite/complicações , Hipersensibilidade a Leite/diagnóstico , Substitutos do Leite Humano , Testes de Irritação da Pele/métodos , Estudos Retrospectivos , Seguimentos , Imunoglobulina E/sangue , Alérgenos/análise , Hipersensibilidade Alimentar/complicações , Hipersensibilidade Alimentar/diagnóstico
12.
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
14.
J Chromatogr A ; 1627: 461396, 2020 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-32823101

RESUMO

Comprehensive two-dimensional gas chromatography (GC×GC) based on flow-modulation (FM) is gaining increasing attention as an alternative to thermal modulation (TM), the recognized GC×GC benchmark, thanks to its lower operational cost and rugged performance. An accessible, rational procedure to perform method translation between the two platforms would be highly valuable to facilitate compatibility and consequently extend the flexibility and applicability of GC×GC. To enable an effective transfer, the methodology needs to ensure preservation of the elution pattern, separation power, and sensitivity. Here, a loop-type thermal modulation system with dual detection (TM-GC×GC-MS/FID) used for the targeted analysis of allergens in fragrances is selected as reference method. Initially, six different columns configurations are systematically evaluated for the flow-modulated counterpart. The set-up providing the most consistent chromatographic separation (20 m x 0.18 mm dc x 0.18 µm df + 1.8 m x 0.18 mm dc x 0.18 µm df) is further evaluated to assess its overall performance in terms of sensitivity, linearity, accuracy, and pattern reliability. The experimental results convincingly show that the method translation procedure is effective and allows successful transfer of the target template metadata. Additionally, the FM-GC×GC-MS/FID system is suitable for challenging applications such as the quantitative profiling of complex fragrance materials.


Assuntos
Cromatografia Gasosa/métodos , Alérgenos/análise , Calibragem , Cromatografia Gasosa/normas , Ionização de Chama , Limite de Detecção , Perfumes/análise , Perfumes/normas , Reprodutibilidade dos Testes
15.
Environ Sci Pollut Res Int ; 27(36): 45447-45459, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32789634

RESUMO

Cumulative data indicate that pollen grains and air pollution reciprocally interact. Climate changes seem also to influence pollen allergenicity. Depending on the plant species and on the pollutant type and concentration, this interaction may modify the features and metabolism of the pollen grain. Previous results revealed a significant positive correlation between pollen and aeroallergen, even using two different samplers. However, some discrepancy days have been also detected with low pollen but high aeroallergen concentrations. The main aim of the present paper is to find how the environmental factors, and specially pollutants, could affect the amount of allergens from olive and grass airborne pollen. Pollen grains were collected by a Hirst-type volumetric spore trap. Aeroallergen was simultaneously sampled by a low-volume Cyclone Burkard sampler. Phl p 5 and Ole e 1 aeroallergen were quantified by double-sandwich ELISA test. The data related to air pollutants, pollen grains, and aeroallergens were analyzed with descriptive statistic. Spearman's correlation test was used to identify potential correlations between these variables. There is a significant positive correlation between aeroallergens and airborne pollen concentrations, in both studied pollen types, so allergen concentrations could be explained with the pollen concentration. The days with unlinked events coincide between olive and grass allergens. Nevertheless, concerning to our results, pollutants do not affect the amount of allergens per pollen. Even if diverse pollutants show an unclear relationship with the allergen concentration, this association seems to be a casual effect of the leading role of some meteorological parameters.


Assuntos
Poluentes Atmosféricos , Poluentes Ambientais , Olea , Poluentes Atmosféricos/análise , Alérgenos/análise , Proteínas de Plantas/análise , Poaceae , Espanha
16.
Food Chem ; 332: 127389, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32645674

RESUMO

Food allergens that cause anaphylactic reactions have become an important health problem worldwide. Among them, shrimp is a popular seafood in many cuisines. The best way to avoid allergic reactions is to mitigate the intake of food allergens. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for the detection of shrimp DNA. Using LAMP primers, the identification of shrimp DNA by the LAMP assay was specific and rapid (within 30 min). It exhibited no cross-reaction with the DNA of other Crustacea, including crabs and lobster, and at least 0.01% shrimp DNA existed in the test sample. Additionally, the sensitivity of LAMP for detecting shrimp DNA was 100-fold greater than that of conventional PCR. LAMP for the detection of shrimp DNA was reproducible regardless of whether the genomic DNA was extracted from boiled, steamed or roasted shrimp samples. In summary, the LAMP assay established herein not only could be potentially used for diagnosing shrimp DNA but could also be applicable for identifying shrimp allergens in commercial food products in marketplaces.


Assuntos
Alérgenos/análise , Técnicas de Amplificação de Ácido Nucleico/métodos , Penaeidae/genética , Alimentos Marinhos/análise , Alérgenos/genética , Animais , Sequência de Bases , Braquiúros/genética , Primers do DNA/metabolismo , Nephropidae/genética , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Alinhamento de Sequência
17.
Food Chem ; 332: 127413, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32652410

RESUMO

Mass spectrometry-based methods coupled with stable isotope dilution have become effective and widely used methods for the detection and quantification of food allergens. Current methods target signature peptides resulting from proteolytic digestion of proteins of the allergenic ingredient. The choice of appropriate stable isotope-labelled internal standard is crucial, given the diversity of encountered food matrices which can affect sample preparation and analysis. We propose the use of concatemer, an artificial and stable isotope-labelled protein composed of several concatenated signature peptides as internal standard. With a comparative analysis of three matrices contaminated with four allergens (egg, milk, peanut, and hazelnut), the concatemer approach was found to offer advantages associated with the use of labelled proteins, ideal but unaffordable, and circumvent certain limitations of traditionally used synthetic peptides as internal standards. Although used in the proteomic field for more than a decade, concatemer strategy has not yet been applied for food analysis.


Assuntos
Alérgenos/análise , Proteômica/métodos , Alérgenos/metabolismo , Sequência de Aminoácidos , Animais , Arachis/metabolismo , Cromatografia Líquida de Alta Pressão , Corylus/metabolismo , Ovos/análise , Marcação por Isótopo , Espectrometria de Massas , Leite/metabolismo , Isótopos de Nitrogênio/química , Peptídeos/análise , Peptídeos/química , Proteômica/normas , Padrões de Referência , Espectrometria de Massas em Tandem
18.
J Environ Sci (China) ; 95: 43-48, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32653191

RESUMO

Pollen pollution and allergy are becoming prominent issues in China. However, few studies on pollinosis have been reported. As an allergen in the atmosphere, allergenic Humulus scandens pollen was collected from four districts of Shanghai, including Wusong (WS), Jiading (JD), Xujiahui (XJH) and Songjiang (SJ). The mass concentrations of SO2, NO2, O3, PM10, and PM2.5 (particulate matter with air dynamic diameter less than 10 and 2.5 µm, respectively) near the four sampling sites were also recorded during Humulus scandens pollen season. The allergenicity of the Humulus scandens pollen was assessed by using of a rat model and enzyme linked immunosorbent assay (ELISA). Relationships between the allergenicity and air pollutants were correlated. Our results demonstrated that the biological viability of the pollens collected from the four districts exhibited no significant differences. ELISA and dot blotting results further demonstrated that the serum of sensitized rats exhibited much higher immune-reactive response than that of control groups. Western blotting showed that the 15 KD (1KD = 1000 dalton) proteins of Humulus pollen led to the allergic response. The allergenic intensity of Humulus pollen protein from different samples followed the pattern: WS > JD > XJ > SJ. There was a negative relationship between the allergenicity of Humulus pollens and PM10 (R = -0.99) / PM2.5 (R = -0.73), and a positive relationship with O3 (R = 0.92). These data clearly showed that PM10 and PM2.5 could enhance Humulus pollen protein release, and O3 could aggravate the allergenicity of the Humulus pollen.


Assuntos
Poluentes Atmosféricos/análise , Poluição do Ar/análise , Humulus/imunologia , Alérgenos/análise , Animais , China , Material Particulado/análise , Pólen/química , Pólen/imunologia , Ratos
19.
Environ Monit Assess ; 192(8): 537, 2020 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-32696086

RESUMO

A thunderstorm is a risk factor for severe respiratory allergy or asthma attacks in patients suffering from pollen/spore allergy. This study aimed to investigate the changes in the spectrum and quantity of pollen and fungal spores in the air of Bratislava during summer storms as well as the impact of selected environmental parameters on these changes. Pollen/spore samples were collected using a Burkard volumetric aerospore trap during summer 2016. To identify those types of pollen/spores that may harm human health during the storm episodes, we analysed how the concentration of individual bioparticles in the air changed during pre-storm/storm/post-storm periods. The effect of environmental variables on the concentration of selected pollen/spore types was evaluated through Spearman's correlation analysis. The results of our study suggest that thunderstorm-related respiratory allergy symptoms in the study area may be caused by (1) spores of Myxomycetes, the airborne concentration of which increases due to an increase in wind speed during the pre-storm period; (2) ruptured pollen and Diatripaceae spores, the concentration of which increases due to increase in precipitation and relative air humidity, respectively, during the storm period; and (3) spores of Fusarium and Leptosphaeria, the concentration of which increases due to increase in precipitation and air temperature, respectively, during the post-storm period.


Assuntos
Microbiologia do Ar , Poluentes Atmosféricos/análise , Alérgenos/análise , Monitoramento Ambiental , Europa (Continente) , Humanos , Estações do Ano , Esporos Fúngicos
20.
Food Chem ; 331: 127276, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32540696

RESUMO

The inclusion on the label of packed foods of any ingredient or technological adjuvant causing allergies is required by EU food legislation. In this study a targeted proteomics method for detecting four allergens in animal-derived food matrices was developed. Liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) was used to select marker peptides from four allergens and develop a quantitative method able to simultaneously detect the presence of milk, egg, crustaceans and soy. The method was validated on fish or swine processed food products contaminated at 5 µg g-1 for milk and egg and 10 µg g-1 for soy and crustaceans. The method was tested by analyzing commercial food products with high protein content and was compared to the ELISA technique. Our results indicated the presence of soy not reported on the food label of some products, pointing out the need for efficient controls to protect allergic consumers.


Assuntos
Alérgenos/análise , Cromatografia Líquida/métodos , Análise de Alimentos/métodos , Hipersensibilidade Alimentar , Espectrometria de Massas em Tandem/métodos , Animais , Ensaio de Imunoadsorção Enzimática , Limite de Detecção , Leite/química , Peptídeos/análise , Carne de Porco/análise , Reprodutibilidade dos Testes , Alimentos Marinhos/análise , Alimentos de Soja/análise , Suínos
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