PURPOSE: The present study was performed to determine the factor, either duration or the temperature of heat treatment, exerting maximal and significant influence on the composition and allergenicity of egg white (EW) proteins. METHODS: Raw EW and 4 kinds of heated EW (fried EW, boiled EW for 10 minutes, boiled EW for 30 minutes, and baked EW for 20 minutes at 170degrees C) were prepared, and subsequently protein extraction was carried out. The proteins were separated by SDS-PAGE, and then immunoglobulin E (IgE) immunoblots were performed with the sera of 7 egg-allergic patients. Furthermore, the antigenic activities of ovalbumin (OVA), ovomucoid (OM), and ovotransferrin (OT) in different EW samples were measured by inhibition enzyme-linked Immuno-sorbent assay (ELISA). RESULTS: In SDS-PAGE analysis, the intensity of the protein band at 45 kD (corresponding to OVA) decreased significantly in boiled EW (30 minutes) and baked EW, but no change was observed in the case of boiled EW for 10 minutes. In IgE immunoblots, the IgE response to 34-50 kD (OM and OVA) in boiled EW for 30 minutes decreased significantly, when compared with raw EW and other heated EWs. In inhibition ELISA, a significant decrease in the OVA antigenic activity was observed in boiled EW for 30 minutes amongst other heated EW samples. However, OM antigenic activity in all kinds of heated EW including boiled EW for 30 minutes did not reduce after heat treatment. The OT antigenic activity nearly disappeared in heated EWs except in the case of boiled EW for 10 minutes. CONCLUSIONS: Amongst 4 kinds of heated EWs, the boiled EW for 30 minutes showed the most significant changes both in composition and reduction in allergenicity. Our results revealed that the duration of heat treatment had more influence on the composition and allergenicity of EW proteins than the temperature.
Conalbumin ; Egg Hypersensitivity ; Egg Proteins ; Egg White ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Hot Temperature ; Humans ; Immunoglobulin E ; Immunoglobulins ; Ovalbumin ; Ovomucin ; Ovum ; Proteins

PURPOSE: Reliable predictors of tolerance to cooked egg in an egg allergic population are not established. We investigated the usefulness of the skin prick test to cooked egg in children with egg allergy. METHODS: We studied 36 children with egg allergy. Skin prick tests (SPTs) for the uncooked or cooked form of egg white and egg yolk, whole egg, ovomucoid (OVM), and ovalbumin (OVA) were performed at diagnosis. The reagents of cooked egg for SPT were prepared by baking for 25 minutes in 200 degree oven. We also examined specific IgE levels to whole egg, egg white, egg yolk, OVM, and OVA. RESULTS: Patients with history of allergic reaction to extensively heated egg showed significantly increased wheal size for cooked egg white (median [interquartile range]), 10.5 [7.0-14.6] vs. 4.2 [0.0-5.6], P<0.001) and OVM (9.6 [7.3-13.8] vs. 5.6 [0.0-7.8], P=0.001) than those without the history. The strongest positive correlation was found between wheal size for cooked egg white and OVM (r=0.788, P<0.001). SPT wheal size for cooked egg white were positively correlated with serum OVM-specific IgE levels (r=0.691, P<0.001). Cutoff value was 7.0 mm in SPT wheal size for cooked egg white, the sensitivity was 73.1% and specificity was 99.0%. SPT for cooked egg white showed significantly higher area under curve than serum egg white specific IgE. CONCLUSION: Our results suggest that SPT to cooked egg white may be useful predictor of allergic reaction to cooked egg. Further investigations will be needed.
Area Under Curve ; Child* ; Diagnosis ; Egg Hypersensitivity* ; Egg Proteins ; Egg White ; Egg Yolk ; Hot Temperature ; Humans ; Hypersensitivity ; Immunoglobulin E ; Indicators and Reagents ; Ovalbumin ; Ovomucin ; Ovum* ; Skin Tests ; Skin*

PURPOSE: Atopic dermatitis is a chronic, inflammatory relapsing skin disease fre- quently seen in children with a history of food allergy. Recently, acute allergic reaction to egg in patients with atopic dermatitis who have never been exposed to egg white protein has been reported. The pattern of antigen-antibody reaction by means of IgE and IgG western blotting was analysed to evaluate the possibility of non- dietary sensitization. METHODS: Total 16 children(<3 years old) with egg allergy were enrolled. Eight patients of them had never ingested egg before(Non-dietary sensitization, NDS) and the other 8 patients had a history of previous egg ingestion without significant clinical symptoms(Dietary sensitization, DS). Egg proteins were analysed by SOS-PAGE and antigen-antibody reaction by means of IgE and IgG was deteded by Western-Blotting. RESULTS: The pattern of IgE antibodies to egg white protein were similar in the two groups. IgG Western blotting to egg white in the two groups, the ovotran sferrine and ovalbumin were significantly bound by 100% of sera, respectively. In DS patients, specific IgG antibodies to ovomucoid was significantly bound by 87% (7/8) of sera whereas only 38%(3/8) of sera had detectable specific IgG antibodies against ovomucoid in NDS patients. CONCLUSION: Theses results suggest that specific IgE to egg white related to exposure of egg white protein regar specific IgG to ovomucoid is related to dietary ingestion of egg. Specific IgG to ovalbumin and ovotrans-ferrine show good clinical correlation, further study might be essential to clarify to this hypothesis.
Allergens* ; Antibodies ; Antigen-Antibody Reactions ; Blotting, Western ; Child ; Dermatitis, Atopic ; Eating ; Egg Hypersensitivity ; Egg Proteins ; Egg White ; Food Hypersensitivity ; Humans ; Hypersensitivity ; Immunoglobulin E* ; Immunoglobulin G* ; Ovalbumin ; Ovomucin ; Ovum* ; Skin Diseases

PURPOSE: Egg (egg white) allergies are among the most common food allergies in infants and young children. Serum egg white-specific IgE (sIgE) levels have been shown to be correlated with clinical symptoms, and the predictive decision point of sIgE levels has been proposed and used widely in the clinical setting. However, some patients whose sIgE levels to egg white are higher than the predictive decision point value show no clinical symptoms, and vice versa. This study was conducted to evaluate the clinical usefulness of sIgE antibodies to egg white and its components in the diagnosis of egg allergies. METHODS: Forty-one patients younger than 2 years of age with no experience of egg intake due to concerns regarding allergies or a non-specific clinical response to eggs were enrolled. Total IgE levels and the levels of IgE antibodies specific for egg white and its components (ovomucoid, ovalbumin, and conalbumin) were measured by ImmunoCAP testing. The clinical response of the subjects was confirmed by an open oral food challenge (OFC). RESULTS: Fifteen (71.4%) out of 21 patients in the egg white-sIgE > or =2 kU/L group showed a positive response, while 10 (50.0%) out of 20 patients in the egg white-sIgE <2 kU/L group showed a negative response to the OFC. There were no statistically significant differences in the levels of sIgE antibodies against egg white and its components between the positive and negative open OFC groups. In addition, there were no statistically significant differences in the levels of sIgE antibodies against egg white and its components based on an intra-group analysis. CONCLUSIONS: Our results show that the sensitivity and specificity of the predictive decision point values for egg white-sIgE antibodies by ImmunoCAP were relatively low in Korean children. In addition, no egg white component predicted the clinical reactivity of the subjects. We suggest that the predictive decision point value for a positive egg oral challenge test by ImmunoCAP should be re-evaluated. Moreover, we suggest that careful personal history recording and challenge tests are necessary for the correct diagnosis of an egg allergy.
Antibodies ; Child ; Egg Hypersensitivity ; Egg Proteins ; Egg White ; Eggs ; Food Hypersensitivity ; Humans ; Hypersensitivity ; Immunoglobulin E ; Infant ; Ovalbumin ; Ovum

PURPOSE: It is known that ovomucoid, an egg allergen, is heat resistant and remains soluble after heating. However, a recent study showed that the antigenic activity of ovomucoid could be reduced by heating when egg white (EW) was mixed with wheat flour. This study was performed to determine the influence of wheat flour on the antigenic activities of EW proteins when EW is heated, and the influence of the duration of heat treatment. METHODS: A mixture of EW and wheat flour was kneaded for 10 minutes and then baked at 180degrees C for 10 minutes and 30 minutes. The EW without wheat flour was also heated at 180degrees C for 10 minutes and 30 minutes. The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and IgE immunoblotting was performed with the pooled sera of 5 egg-allergic patients. The antigenic activities of ovomucoid in different EW samples were measured by inhibition enzyme-linked immunosorbent assay (ELISA). RESULTS: 1) SDS-PAGE: the intensity of the 37-50 kD bands (overlapped bands of ovomucoid and ovalbumin) decreased significantly in the mixture of EW and wheat flour baked for 30 minutes, compared with the mixture baked for 10 minutes, heated EW and raw EW. 2) IgE immunoblot: in the mixture of EW and wheat, a remarkable decrease of IgE reactivity to 37-50 kD was observed when baked for 30 minutes. 3) Inhibition ELISA: the antigenic activity of ovomucoid decreased significantly in the mixture of EW and wheat baked for 30 minutes, but not in the heated pure EW. CONCLUSIONS: This study showed that the antigenic activity of ovomucoid can be reduced by baking EW with wheat flour. The decrease in ovomucoid antigenicity in the baked mixture of EW and wheat flour was dependent on the time of heat treatment, indicating that heating should be prolonged to achieve a reduction in ovomucoid antigenic activity.
Acrylic Resins ; Egg Proteins ; Egg White ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Flour ; Heating ; Hot Temperature ; Humans ; Immunoblotting ; Immunoglobulin E ; Ovomucin ; Ovum ; Proteins ; Sodium Dodecyl Sulfate ; Triticum

Hemo-Q (R) and Albumax (R) solution are widely used antianemics. Hemo-Q (R) contains cow's milk protein (casein) and Albumax (R) contains egg white protein (ovalbumin). Cow's milk protein and egg protein can cause common allergic diseases in infants and young children. We reported two cases of young children with milk and egg allergy who presented skin symptoms after ingestion or cutaneous contact with Hemo-Q (R) or Albumax (R) solution. When Hemo-Q (R) or Albumax (R) solution was taken or rubbed on, erythematous papules and wheals were developed in 10-20 minutes, but other antianemics which don't contain milk or egg protein contents didn't show skin manifestations.
Caseins ; Child ; Eating ; Egg Hypersensitivity ; Egg White* ; Food Hypersensitivity ; Humans ; Infant ; Milk Proteins* ; Milk* ; Ovalbumin ; Ovum* ; Skin ; Skin Manifestations ; Urticaria*

OBJECTIVETo study the influence of heat-induced epitope retrieval (HIER) on endogenous avidin-binding activity (EABA) and to establish an effective way to block EABA in immunohistochemistry.

METHODSSystematically screening EABA in 164 (679 samples) formalin-fixed and paraffin-embedded human tissues including 76 (102 samples) normal tissues and 88 (577 samples) tumor tissues as well as 4 (80 samples) formalin-fixed and paraffin-embedded rat normal tissues using tissue array (tissue chip), HIER, immunohistochemistry and egg white solution blocking. In addition, EABA was also examined in 9 (15 samples) human frozen tissues.

RESULTS(1) EABA was detected in frozen tissues. (2) No staining for EABA was seen in formalin-fixed and paraffin-embedded tissues. (3) EABA was revealed after the tissues treated with microwave HIER. (4) The density of signal for EABA was variable from tissue to tissue and cell to cell. (5) The signals of EABA expressed in scatter or diffuse in tissues and in granular form in cytoplasm. (6) EABA was found in a wide range of epithelial tissues, especially in gland epithelia of normal and tumor tissues. These included kidney, adrenal cortex, liver, C cells of thyroid gland, oxyphil cells of parathyroid, fundal gland of stomach, sebaceous gland of skin, duct of salivary; oncocytoma and papillary adenocarcinoma of kidney and thyroid gland, adenolymphoma of parotid, carcinoma of liver cell, adenocarcinoma of stomach, colon, prostate, gall bladder and endometrium, and so on. (7) EABA was easier revealed by higher pH value buffer (EGTA pH 9.0) than that with lower pH value (EDTA pH 8.0 and citrate pH 6.0). (8) The revealed EABA could be effectively blocked using 20% egg white solution.

CONCLUSIONSHIER could unmask EABA in formalin-fixed and paraffin-embedded tissues. The unmasked EABA present in a wide range of human normal and tumor tissues as well as in rat normal tissues. The EABA could influence routine immunohistochemistry staining when using (strept)avidin-horseradish peroxidase detective system. The egg white solution could effectively block EABA and eliminate the influence of EABA on immunohistochemistry.

Animals ; Avidin ; antagonists & inhibitors ; metabolism ; Biotin ; metabolism ; Cells, Cultured ; Egg Proteins ; pharmacology ; Epithelial Cells ; metabolism ; Epitopes ; Female ; Hot Temperature ; Humans ; Immunohistochemistry ; Male ; Neoplasms ; metabolism ; pathology ; Rats

No abstract available.
Avidin*

OBJECTIVETo measure the prevalence, the possible causes and the influencing factors of allergy, food hypersensitivity and food intolerance in 0-36 month old infants in 8 cities in China.

METHODTotally 2632 infants from the outpatient departments of prevention and health care of two representative hospitals in 8 Chinese cities were randomly selected by applying multistage cluster sampling method from October 2011 to March 2012, and a one-on-one survey to infants' parents was conducted to investigate infants' sensitization status.

RESULTSelf-reported infant allergy rate was 17.97% (473/2632) ; self-reported food hypersensitivity and food intolerance rates were 6.53% (172/2632) and 4.26% (112/2632) , respectively. The proportion of self-reported food hypersensitivity of 0-12 months old infants was 4.47% (74/1656) and their top five allergens in a descending order were eggs (28.38%) , shrimp (25.68%) , fish (21.62%) , milk (18.92%) and wheat (4.05%) . The proportion of self-reported 13-36 months old infant's food hypersensitivity was 10.05% (98/976) . The top five allergens were shrimp (33.93%) , fish (26.79%) , eggs (23.21%) , milk (12.50%) and soy (3.57%) in 13-24 months group, while fish (38.24%) , shrimp (35.29%) , eggs (20.59%) , milk (20.59%) and peanuts (2.94%) in 25-36 months group. Both 7-12 and 13-24 month old were the highest incidence (both of them were 11.98%, 58/484) of age for developing food hypersensitivity and 7-12-month old was also the highest incidence (8.47%, 41/484) of age for food intolerance. The self-reported food intolerance rate was 3.68% (61/1656) and 5.23% (51/976) in the two age groups, respectively. Age, parental history of allergy and father's educational level (OR was 2.452, 1.482 and 2.598, respectively, P < 0.01) were the risk factors of food hypersensitivity; within two weeks of sickness (OR = 1.267, P < 0.05) was the risk factor of food intolerance.

CONCLUSIONInfancy was the most vulnerable period of life of getting allergy, therefore, it is necessary for all infants to prevent allergy through a variety of effective strategies.

Age Distribution ; Child, Preschool ; China ; epidemiology ; Dietary Proteins ; adverse effects ; Egg Hypersensitivity ; epidemiology ; etiology ; Eggs ; adverse effects ; Female ; Food Hypersensitivity ; epidemiology ; etiology ; prevention & control ; Humans ; Incidence ; Infant ; Infant Food ; adverse effects ; Infant Formula ; Infant, Newborn ; Male ; Milk Hypersensitivity ; epidemiology ; etiology ; prevention & control ; Risk Factors ; Sampling Studies ; Sex Distribution ; Surveys and Questionnaires

PURPOSE: Serum screening test to detect specific immunoglobulin E (IgE) is an important step for the assessment of potential allergenicity of genetically modified (GM) food. The purpose of this study was to evaluate the usefulness of pooled serum for serum screening instead of individual serum. METHODS: Children with allergic disease were recruited and those who were sensitized to peanut or egg white were selected to obtain their sera. Sensitization to these foods was determined when the level of specific IgE was over 0.35 kU/L by ImmunoCAP. The patients were divided into subgroups according to their level of specific IgE. Raw proteins were extracted and immunoblot analysis was performed to compare the immunoreactivity between individual serum and pooled serum. RESULTS: Pooled serum from peanut-sensitized allergic children showed all the bands which were shown in immunoblot analysis by using individual serum and peanut protein extract. These findings were demonstrated both in pooled serum with low level of peanut-specific IgE and in those with high level of peanut-specific IgE. Likewise, there was no difference in the immunoreactivity between individual serum and pooled serum from egg white-sensitized allergic children. CONCLUSION: Pooled serum can be used as an alternative to individual serum for the serum screening in the allergenicity assessment of GM food.
Child ; Egg White ; Food, Genetically Modified ; Humans ; Immunoglobulin E ; Immunoglobulins ; Mass Screening ; Ovum ; Proteins