Biomedical subjects

Soheila J Maleki

Publications and source records attributed to Soheila J Maleki.

11 recordsLinked to original sources

PubMed2004

Allergenic properties of roasted peanut allergens may be reduced by peroxidase.

Peanut allergy is a public health issue. The culprits are the peanut allergens. Reducing the allergenic properties of these allergens or proteins will be beneficial to allergic individuals. In this study, the objective was to determine if peroxidase (POD), which catalyzes protein cross-linking, reduces the allergenic properties of peanut allergens. In the experiments, protein extracts from raw and roasted defatted peanut meals at pH 8 were incubated with and without POD in the presence of hydrogen peroxide at 37 degrees C for 60 min. The POD-treated and untreated samples were then analyzed by SDS-PAGE, western blots, and competitive inhibition ELISA. IgE binding or allergenicity was determined in blots and ELISA. Results showed that POD treatment had no effect on raw peanuts with respect to protein cross-linking. However, a significant decrease was seen in the levels of the major allergens, Ara h 1 and Ara h 2, in roasted peanuts after POD treatment. Also, polymers were formed. Despite this, a reduction in IgE binding was observed. It was concluded that POD induced the cross-linking of mainly Ara h 1 and Ara h 2 from roasted peanuts and that, due to POD treatment, IgE binding was reduced. The finding indicates that POD can help reduce the allergenic properties of roasted peanut allergens.

Allergens↗

PubMed2004

cDNA clone of a putative peanut (Arachis hypogaea L.) trypsin inhibitor has homology with peanut allergens Ara h 3 and Ara h 4.

Trypsin inhibitors are pathogenesis-related (PR) proteins, which play an important role in the plant defense mechanism against insects and pathogens. Peanut trypsin inhibitors are low molecular mass seed storage proteins. Like peanut allergens, they are stable to acid and heat, resistant to digestion, and can have a negative impact on human health. In peanut, five Bowman-Birk trypsin inhibitors (BBTI) have been isolated and amino acid sequences published. However, to date, no peanut BBTI sequence is available at both the cDNA and the genomic levels. The objectives of this investigation were (i) to synthesize degenerate oligonucleotides based on conserved regions of published amino acid sequences of BBTI, BII, and BIII; (ii) to isolate, sequence, and analyze at least one positive peanut trypsin inhibitor cDNA clone using the synthesized (32)P-labeled oligonucleotides as probes; and (iii) to determine its trypsin inhibitory activity. Thirty-two degenerate oligonucleotides DNA primers of 24 nucleotides each were synthesized based on the published amino acid sequences of peanut BBTI, and two were selected as probes to screen a peanut Lambda gt 11 cDNA library. Three putative positive clones were isolated, purified, and subcloned, and one was sequenced. Sequence analysis revealed a partial cDNA clone of 643 bp with a start codon. This clone shares 93 and 96% nucleotide sequence homology with peanut allergens Ara h 3 and Ara h 4 cDNA clones, respectively. A trypsin inhibitor assay revealed that peanut allergen Ara h 3 has a trypsin inhibitory activity of 11 238 TIA/mg protein. We concluded that peanut allergen Ara h 3 may also function as a trypsin inhibitor.

Allergens↗

PubMed2004

Food processing: effects on allergenicity.

PURPOSE OF REVIEW: There has been an increase in the prevalence of food allergy, asthma and severe reactions to foods in the past decade. The reason for this increase is unknown and despite the potential for a lethal outcome, no treatments or therapies are available. Many areas are being explored to understand the increase in food allergy and the role of modern processing techniques is one of them. RECENT FINDINGS: The fact that processing influences allergenicity is not a new concept, but due to some definitive studies in the last few years the topic has been revitalized. Not only is the processed form of a particular allergen important in assessment of allergenicity, but the role of the food matrix in which an allergen is processed must also be evaluated. The issue of allergenicity of particular foods is therefore highly complex, and even with our existing knowledge we have just begun to understand some of the contributions of processing to allergenic properties of proteins and foods. SUMMARY: Researchers and medical personnel have begun to weigh the benefits and potential disadvantages of assessing the allergenicity of foods in unprocessed forms. In this review, the known effects of processing on the allergenicity of various foods will be discussed along with the implications of the most recent studies on the future direction of research in this area.

Allergens↗

PubMed2004

Staphylococcus aureus AgrA binding to the RNAIII-agr regulatory region.

The control of virulence gene expression in the human pathogen Staphylococcus aureus is under the partial control of the two-component quorum-sensing system encoded by genes of the agr locus. The product of the agrA gene has been shown by amino acid sequence similarity to be the putative response regulator; however, binding of AgrA to promoters under its control has not yet been demonstrated. In this study, we isolated and purified soluble AgrA by expression under osmotic shock conditions and ion-exchange chromatography. Purified AgrA showed high-affinity binding to the RNAIII-agr intergenic region by electrophoretic mobility shift assays. Binding was localized by DNase I protection assays to a pair of direct repeats in the P2 and P3 promoter regions of the agr locus. We found that this binding was enhanced by the addition of the small phosphoryl donor, acetyl phosphate. The difference in binding affinity between these two promoters was found to result from a 2-bp difference between the downstream direct repeats of the P2 and P3 sites. Mutation of these base pairs in the P3 site to match those found in the P2 site increased the affinity of AgrA for the P3 site relative to that for the P2 site. These results are consistent with the function of AgrA as a response regulator with recognition sites in the promoter regions of RNAIII and the agr locus.

Bacterial Proteins↗

PubMed2003

Linking peanut allergenicity to the processes of maturation, curing, and roasting.

The processes of peanut maturation, curing, and roasting are known to have an important role in peanut flavors. One of these processes (i.e., roasting) has been found to have an effect on allergenicity. To determine if the other processes (i.e., maturation and curing) affect allergenicity, mature and immature roasted peanuts and peanuts cured at different temperatures (35-77 degrees C) were, respectively, tested for IgE binding and advanced glycation end adducts (AGEs). Peanuts with and without stress proteins, which are associated with peanut maturation and curing, were also tested. Results showed that mature roasted peanuts exhibited a higher IgE binding and AGEs level than immature roasted peanuts. Curing temperatures between 35 and 60 degrees C gave no difference in the profiles. However, a higher curing temperature (i.e., 77 degrees C) exhibited a profile of higher levels of AGEs and IgE binding. These levels were higher in peanuts with stress proteins than without stress proteins. Roasting increased stress protein level and IgE binding. From these results, the processes of maturation and curing, in conjunction with roasting, may be associated with allergenicity, suggesting that these processes may lead to changes in the allergenic properties of peanuts.

Allergens↗

PubMed2003

The major peanut allergen, Ara h 2, functions as a trypsin inhibitor, and roasting enhances this function.

BACKGROUND: The widespread use of peanut products, the severity of the symptoms, and its persistence in afflicted individuals has made peanut allergy a major health concern in western countries such as the United States, United Kingdom, and Canada. In a previous study, the authors showed that the allergenic properties of peanut proteins are enhanced as a result of thermal processing. OBJECTIVE: The purpose of this investigation was to determine whether any specific functions are associated with the major peanut allergen, Ara h 2, and whether the functionality of this protein is influenced by processing. An assay was developed and used to assess structure/function changes in Ara h 2 induced by roasting and the effect of these alterations on the allergenic properties of this major peanut allergen. METHODS: A protein domain homology search was used to determine possible functions for Ara h 2. One of the putative functions (protease inhibition) was tested by means of appropriate enzyme assays and protein gel electrophoresis. Circular dichroism was used to compare the structural properties of Ara h 2 purified from raw and roasted peanuts. RESULTS: Ara h 2 purified from peanuts is homologous to and functions as a trypsin inhibitor. Roasting caused a 3.6-fold increase in trypsin inhibitory activity. Functional and structural comparison of the Ara h 2 purified from roasted peanuts to native and reduced Ara h 2 from raw peanuts revealed that the roasted Ara h 2 mimics the behavior of native Ara h 2 in a partially reduced form. CONCLUSIONS: The data indicate that thermal processing might play an important role in enhancing the allergenic properties of peanuts. Not only has it previously been shown to affect the structural and allergic properties of peanut proteins but also, for the first time, the functional characteristics of an allergen. These structural and functional alterations are likely to influence the allergenicity of peanuts.

2S Albumins, Plant↗

PubMed2003

Characterization of lymphocyte responses to peanuts in normal children, peanut-allergic children, and allergic children who acquired tolerance to peanuts.

Comparing lymphocyte responses to allergenic and nonallergenic foods could reveal the differences between pathogenic and normal immune responses to foods. Defining the cytokine-producing phenotypes of peanut-specific lymphocytes from peanut-allergic children, children who outgrew peanut allergy, and children who have always tolerated peanuts may be useful for understanding the mechanisms of food tolerance. Investigating immune responses against foods is hindered, however, by the fact that circulating food antigen-specific lymphocytes are very rare. In a novel approach we used carboxyfluorescein succinimidyl ester to detect peanut-specific lymphocytes by flow cytometry. We confirmed that these cells are indeed peanut specific by cloning. Peanut-allergic donors show Th2 polarization of cytokine production by peanut-specific cells (IFN-gamma (low), TNF-alpha (low), IL-4 (high), IL-5 (high), IL-13 (high)). Conversely, nonallergic children and children who have outgrown their allergy show Th1 skewing to peanut antigens (IFN-gamma(high), TNF-alpha (high), IL-4 (low), IL-5 (low), IL-13(low)), similarly to nonallergenic food antigens (beta-lactoglobulin, OVA). This finding suggests that peanut antigens do not intrinsically induce Th2 skewing, but that the type of response depends upon the donor's allergic status. In conclusion, food allergic status is characterized by a Th2 response whereas Th1-skewed responses underlie oral tolerance.

Adolescent↗

PubMed2002

Analysis of the DNA-binding properties of MyoD, myogenin, and E12 by fluorescence anisotropy.

MyoD and Myogenin are dominant myogenic regulatory factors (MRFs), which are involved in control of muscle-specific gene expression. The ubiquitously expressed E12 dimerizes with MyoD and Myogenin and has been shown to enhance their DNA-binding and transcriptional activities. In this study, fluorescence anisotropy assays have been used to determine the Gibb's free energy of dissociation (DeltaG) for MyoD, Myogenin, and E12 as homo- and heterodimers to the well-characterized myosin light chain enhancer (MLC), muscle creatine kinase (MCK) enhancer, and mutant thereof. The heterodimers of MyoD or Myogenin with E12 bound the MCK enhancer equally well (DeltaG = 21 kcal/mol). The homodimers varied dramatically in both MLC and MCK enhancer binding affinity. MyoD homodimer bound the MCK enhancer with the highest affinity (DeltaG = 19.6 kcal/mol) in comparison with the Myogenin homodimer-MCK interaction (DeltaG = 16.6 kcal/mol) and E12 homodimer-MCK interaction (DeltaG = 18.0 kcal/mol). The slope and shape of the binding isotherms revealed that with the exception of the E12 homodimer-MCK enhancer interaction, the other proteins bound with high levels of positive cooperativity. In contrast, the E12 homodimer-MCK enhancer interaction actually occurs with significant negative cooperativity. The binding of these proteins to MLC enhancer mimicked binding to the MCK enhancer, but with much lower affinities. These data support the hypothesis that DNA acts as an allosteric ligand facilitating the dimerization of these proteins. The combination of differential affinity and cooperativity explains why the heterodimers are the active species in transcriptional regulation.

Creatine Kinase↗

PubMedDate not supplied

Structural and functional alterations in major peanut allergens caused by thermal processing.

The majority of foods that we eat are subjected to some type of processing either at home or by the manufacturer. The biochemical reactions that occur in foods as a result of thermal processing can be both beneficial and harmful. Here, we briefly review the effects of thermal processing and some of the effects of the Maillard reaction on the allergenicity of food proteins. Specifically, we focus on the known effects of roasting on the allergenic properties of peanut proteins and the contribution of Maillard reaction products or advanced glycation end products to these observed effects. The most thorough understanding of the effects of thermal processing on allergenicity involves the peanut proteins. Thermal processing alters specific biophysical and immunological properties of peanut proteins, such as structure, function, solubility, digestibility, immunoglobulin E (IgE) binding, and T-cell responses. A better understanding of the effects of thermal processing-induced biochemical and immunological alterations is of utmost importance for proper risk assessment of existing and newly introduced proteins in the food source, as well as development of effective diagnostic tools and therapeutic treatments for food allergy.

Allergens↗

PubMedDate not supplied

Can commercial peanut assay kits detect peanut allergens?

Peanut is the food group mostly associated with severe and fatal allergic reactions. In the United States, more than 90% of peanut-allergic individuals' serum IgE recognized peanut proteins Ara h 1 and Ara h 2, thus establishing these proteins as major peanut allergens. The amount of Ara h 1 and Ara h 2 in 3 varieties of peanut cultivars that are commonly processed in the industrialized countries was determined to be 12-16 and 6-9%, respectively. Current commercial peanut test kits use polyclonal peanut-specific antibodies to detect soluble or buffer extractable peanut proteins. Because the 2 major peanut allergens Ara h 1 and Ara h 2 are isolated from soluble peanut proteins, it is generally assumed that these commercial kits can detect peanut allergens, although none of these kits claims to detect peanut allergen. This study showed for the first time that the peanut test kits could, in fact, detect major peanut allergens Ara h 1 and Ara h 2 in both native or heat-denatured structures; therefore, these kits qualified to be classified as peanut allergen enzyme-linked immunosorbent assays.

2S Albumins, Plant↗

PubMedDate not supplied

Competitive inhibition ELISA for quantification of Ara h 1 and Ara h 2, the major allergens of peanuts.

Allergies to peanuts are becoming an increasingly important health problem as a result of the persistence and severity of the reaction in allergic individuals. Because no treatment currently is available, avoidance is the only option for peanut-allergic individuals. Avoidance of an abundant and often disguised food such as peanuts, however, is very difficult; therefore, competitive inhibition ELISAs were developed to detect and quantitate each of the major peanut allergens, Ara h 1 and Ara h 2. Under optimal conditions for each assay, the sensitivity of the Ara h 1 and Ara h 2 detection assays were 12 and 0.5 ng/mL, respectively. These assays were primarily devised to effectively compare the levels of Ara h 1 and Ara h 2 in a wide variety of peanuts or peanut products but can also be used to identify cross-reactive antigens. The method is simple and rapid, requiring only one allergen-specific antibody and, therefore, could be adapted specifically to detect the presence of these individual allergens in different foods.

2S Albumins, Plant↗