Immunoglobulin E-binding Proteins of Cooked Walnuts in Korean Children

PURPOSE: The immunological characteristics of young Korean children with walnut (WN) allergy and the influence of different cooking methods on WN proteins have not been evaluated to date. This study aimed to evaluate the major WN allergens identified among Korean children, together with changes in WN antigenicity caused by common cooking methods. METHODS: We enrolled children under the age of 13 years with WN serum-specific immunoglobulin (Ig) E concentrations. The protein fractions of dry-fried and boiled WN extracts were compared with those of raw WNs using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), 2-dimentional gel electrophoresis (2DE) and a proteomic analysis using electrospray ionization (liquid chromatography-mass spectrometry [LC-MS]). An immunoblotting analysis was conducted to examine IgE reactivity toward raw WNs using serum samples from 6 children with a clinical WN allergy. To determine the processed WN proteins with IgE-binding capacity, a 2D-immunoblotting analysis was performed using the pooled sera of 20 WN-sensitized children. RESULTS: Protein bands from raw WNs were identified at 9, 16, 28, 52, 58, and 64 kDa via SDS-PAGE. The 9- and 16-kDa protein bands were enhanced by boiling, whereas the 52- and 64-kDa bands were considerably diminished. On LC-MS analysis, of the 66 IgE-binding proteins present in raw WNs, 57 were found in dry-fried WNs, but only 4 in boiled WNs. The sera of 5 out of 6 participants reacted with the 52-kDa protein bands and those of 4 out of 6 participants reacted with the 16- and 28-kDa protein bands, respectively. Meanwhile, a 2D-immunoblotting result confirmed the presence of different binding patterns among children who consumed cooked WNs. CONCLUSIONS: The protein profile of boiled WNs is substantially different from that of raw WNs. However, 4 proteins including prolamins remained stable after dry-frying or boiling. Further studies are needed to evaluate the clinical relevance of these findings.

Gastroprotective Effects of Glutinous Rice Extract against Ethanol-, Indomethacin-, and Stress-induced Ulcers in Rats / 전남의대학술지

This study was designed to evaluate the efficacy of an orally administered aqueous extract of glutinous rice (GRE) to protect against acute gastric mucosal lesions induced by ethanol, indomethacin, and water immersion restraint stress in rats and to characterize the active substances responsible for the protection. GRE was shown to dose-dependently prevent the gastric lesions induced by the above ulcerogenic treatments at doses of 30 to 300 mg/kg. GRE treatment increased the gastric mucin content and partially blocked the ethanol-induced depletion of the gastric mucus layer. Also, it increased the nonprotein sulfhydryl concentration in the gastric mucosa. The gastroprotective action of GRE was markedly enhanced by co-treatment with 4-8 mg/kg tea extracts. The activity of GRE was completely lost by heat treatment at 80degrees C for 3 min or treatment with 0.01% pepsin at 37degrees C for 1 h. Protein extraction studies indicated that prolamins are involved in the gastroprotective activity of GRE. Our results suggest that glutinous rice proteins are useful for the prevention and treatment of gastritis and peptic ulcer.

Gastroprotective Effects of Glutinous Rice Extract against Ethanol-, Indomethacin-, and Stress-induced Ulcers in Rats / 전남의대학술지

This study was designed to evaluate the efficacy of an orally administered aqueous extract of glutinous rice (GRE) to protect against acute gastric mucosal lesions induced by ethanol, indomethacin, and water immersion restraint stress in rats and to characterize the active substances responsible for the protection. GRE was shown to dose-dependently prevent the gastric lesions induced by the above ulcerogenic treatments at doses of 30 to 300 mg/kg. GRE treatment increased the gastric mucin content and partially blocked the ethanol-induced depletion of the gastric mucus layer. Also, it increased the nonprotein sulfhydryl concentration in the gastric mucosa. The gastroprotective action of GRE was markedly enhanced by co-treatment with 4-8 mg/kg tea extracts. The activity of GRE was completely lost by heat treatment at 80degrees C for 3 min or treatment with 0.01% pepsin at 37degrees C for 1 h. Protein extraction studies indicated that prolamins are involved in the gastroprotective activity of GRE. Our results suggest that glutinous rice proteins are useful for the prevention and treatment of gastritis and peptic ulcer.

Diagnosis and Management of Grain-Induced Asthma

Grain-induced asthma is a frequent occupational allergic disease mainly caused by inhalation of cereal flour or powder. The main professions affected are bakers, confectioners, pastry factory workers, millers, farmers, and cereal handlers. This disorder is usually due to an IgE-mediated allergic response to inhalation of cereal flour proteins. The major causative allergens of grain-related asthma are proteins derived from wheat, rye and barley flour, although baking additives, such as fungal alpha-amylase are also important. This review deals with the current diagnosis and treatment of grain-induced asthma, emphasizing the role of cereal allergens as molecular tools to enhance diagnosis and management of this disorder. Asthma-like symptoms caused by endotoxin exposure among grain workers are beyond the scope of this review. Progress is being made in the characterization of grain and bakery allergens, particularly cereal-derived allergens, as well as in the standardization of allergy tests. Salt-soluble proteins (albumins plus globulins), particularly members of the alpha-amylase/trypsin inhibitor family, thioredoxins, peroxidase, lipid transfer protein and other soluble enzymes show the strongest IgE reactivities in wheat flour. In addition, prolamins (not extractable by salt solutions) have also been claimed as potential allergens. However, the large variability of IgE-binding patterns of cereal proteins among patients with grain-induced asthma, together with the great differences in the concentrations of potential allergens observed in commercial cereal extracts used for diagnosis, highlight the necessity to standardize and improve the diagnostic tools. Removal from exposure to the offending agents is the cornerstone of the management of grain-induced asthma. The availability of purified allergens should be very helpful for a more refined diagnosis, and new immunomodulatory treatments, including allergen immunotherapy and biological drugs, should aid in the management of patients with this disorder.

Diagnosis and Management of Grain-Induced Asthma

Grain-induced asthma is a frequent occupational allergic disease mainly caused by inhalation of cereal flour or powder. The main professions affected are bakers, confectioners, pastry factory workers, millers, farmers, and cereal handlers. This disorder is usually due to an IgE-mediated allergic response to inhalation of cereal flour proteins. The major causative allergens of grain-related asthma are proteins derived from wheat, rye and barley flour, although baking additives, such as fungal alpha-amylase are also important. This review deals with the current diagnosis and treatment of grain-induced asthma, emphasizing the role of cereal allergens as molecular tools to enhance diagnosis and management of this disorder. Asthma-like symptoms caused by endotoxin exposure among grain workers are beyond the scope of this review. Progress is being made in the characterization of grain and bakery allergens, particularly cereal-derived allergens, as well as in the standardization of allergy tests. Salt-soluble proteins (albumins plus globulins), particularly members of the alpha-amylase/trypsin inhibitor family, thioredoxins, peroxidase, lipid transfer protein and other soluble enzymes show the strongest IgE reactivities in wheat flour. In addition, prolamins (not extractable by salt solutions) have also been claimed as potential allergens. However, the large variability of IgE-binding patterns of cereal proteins among patients with grain-induced asthma, together with the great differences in the concentrations of potential allergens observed in commercial cereal extracts used for diagnosis, highlight the necessity to standardize and improve the diagnostic tools. Removal from exposure to the offending agents is the cornerstone of the management of grain-induced asthma. The availability of purified allergens should be very helpful for a more refined diagnosis, and new immunomodulatory treatments, including allergen immunotherapy and biological drugs, should aid in the management of patients with this disorder.

Fraccionamiento y caracterización de las proteínas solubles de la harina de nuez de Barinas (caryodendron orinocense K. ) / Fractionation and characterization of soluble proteins from nuez de Barinas (caryodendron orinocense K .) flour

La harina de nuez de Barinas (Caryodendron orinocense K.) de acuerdo a estudios realizados presenta un contenido de pro teínas en el rango de 15-18%, que permite se pueda considerar como fuente de proteína. Sin embargo no se conoce la composición de estas proteínas. En este estudio se realizó un proceso de fraccionamiento de acuerdo a la solubilidad en diferentes solventes (agua, cloruro de sodio al 5%, hidróxido de sodio 0,02N, etanol al 70%), y el establecimiento de la composición proteica según el peso molecular de la harina de nuez de Barinas obtenida a partir de las nueces secas, molidas y des grasadas, determinando el rango de pesos moleculares por SDS-PAGE y su comparación con el rango de pesos moleculares de las proteínas de la harina de soya (Glycine max). Asi mismo, se determinó la digestibilidad in vitro por hidrólisis enzimática. Los resultados indican un rango de PM de 20.000- 97.000 daltons para la soya y de 6.500-45.000 daltons para la nuez de Barinas. El peso molecular más bajo sugiere una más fácil digestión de las proteínas, que corrobora el valor de la di - ges tibilidad obtenido de 75%. Las proteínas presentes están cons tituidas por albúminas 50,72%, globulinas 15,56%, prolaminas 23,10% y glutelinas 2,52% que representan el 92,15% del total de proteínas presentes. Estas fracciones presentaron una pureza de 81,57%, 94,01%, 70,86% y 92,53%, respectivamente. Estos resultados sugieren la posibilidad de uso de la harina de Caryodendron orinocense K en el desarrollo de productos alimenticios para niños y ancianos.

Nuez de Barinas (Caryodendron orinocense K.) flour has been reported to have a content of proteins in the range of 15-18% that allows considering it as a source of proteins. However, there is no evidence on the composition of these proteins. The objective of this study was the extraction and separation of the different fractions of proteins based on their solubility in different solvents (water, 5% sodium chloride, sodium hydroxide 0.02N, and 70% ethanol), and the protein composition assessment of the nuez de Barinas flour from dried, milled, and defatted nuts; by analysis of the range of mo lecular weight by SDS-PAGE. Molecular weights of proteins from soybean (Glycine max) flour, were used for comparison. In vitro protein digestibility was determined by enzymatic hydrolysis. Results presented soy proteins in the range of 20.000-97.000 daltons while the nuez de Barinas flour proteins were in the range of 6.500-45.000 daltons. Low molecular weight proteins should suggest a much easier digestibility of these proteins, which is related to the digestibility index of 75% found. Protein composition was found to be 50,72% albumins, 15,56% globulins, 23,10% prolamins, and 2,52% glutelins which represent 92,15% of total proteins present. These fractions showed a percent purity of 81.57, 94.01, 70.86 and 92.53, respectively. These results suggest the possibility to use this flour Caryodendron orinocense K. in developing food products for children and elderly persons.

Diversity and stability study on rice mutants induced in space environment / 基因组蛋白质组与生物信息学报·英文版

To further study the characteristics of changes on the molecular level of rice mutants induced in space environment, we analyzed proteins in leaves and seeds of four rice mutants (two high-tillering and two low-tillering) in the 8(th) and 9(th) generations after a 15-day spaceflight, and compared with their ground controls by two-dimentional polyacrylamide gel electrophoresis and reverse phase liquid chromatography (RPLC). In addition, the albumin, globulin, prolamine, glutelin, and amylose of the mutant seeds were analyzed by RPLC and ultra-violet spectrometry. The results showed that the low-abundance proteins of leaves in the peak tillering stage are more likely to be induced compared with their corresponding controls. The albumin, globulin, and prolamine of the mutant seeds revealed changes when compared with their controls, and the characteristics of changes in different mutants were stably inherited in the 8(th) and 9(th) generations, suggesting that they can be used as bio markers to identity the mutants induced by spaceflight. Moreover, two proteins (SSP9111 and SSP6302) were found to be expressed with high intensity (two-fold change) in different mutants, which were both correlated with photosystem according to mass spectrometry and database searching.