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[Effects of soil moisture on the compensation effect of winter wheat with its partial roots cut off at returning green stage].

The study with pot experiment showed that after cutting partial roots at returning green stage, the growth of winter wheat was restrained at early growth stage, and the leaf area was decreased significantly from returning green to jointing stage but restored to the level of the control at flowering stage. Under high soil moisture condition, root cutting increased the values of chlorophyll fluorescence parameters ETR, phiPS II , qp and qn, at jointing stage significantly. The accumulated dry matter weight per stem after anthesis was significantly higher in root-cut wheat (0. 81 g) than in the control (0. 56 g) , with the accumulation coefficient (AC) of root-cut wheat increased by 38. 79% , but no significant difference was observed in root weight. Under low soil moisture condition, there were no significant differences in the values of chlorophyll fluorescence parameters and accumulated dry matter weight per stem after anthesis between root-cut wheat and the control, but the root weight of root-cut wheat decreased significantly. Soil moisture didn' t enhance the compensation effect of the aboveground biomass and grain yield of root-cut wheat. Root cutting reduced the water consumption of winter wheat significantly. Under high soil moisture condition, root-cut wheat saved 2 000 ml of water, and its water use efficiency (WUE) ( 1. 97 g x kg (-1)) was significantly higher than that of the control (1.70 g x kg(-1)). Under low soil moisture condition, root-cut wheat saved 1500 ml of water, but there was no significant difference in the WUE between root-cut wheat and the control.

Adaptation, Physiological↗

[The dot-blot technic for quantitative determination of wheat protein in sausage].

A simple immunological method is described by which it is possible to detect the wheat flour according to wheat protein in the heat-processed meat products. The wheat proteins were extracted with a buffer TRIS-NaCl (0.02 mol/l, 0.5 mol/l) from: a) wheat flour; b) model meat products with addition of wheat flour (3.2, 3.2, 3.6, 0.8, 0.4 and 0.2%); c) commercial meat products containing wheat flour (sausages-S, P, PF). The aliquots of the extracts were subjected to an immunological dot-blot test. The extracts from the wheat flour and the model meat products were used for calibration of this method. The quantitative evaluation of the test was based on the spectrophotometric determination of the quantity of the violet stain on the nitrocellulose (NC) strip surfaces. The strips were immersed into the acetone and the absorbance were determined at 580 nm. The screening of the samples by dot-blot procedure proved to be an efficient way of the wheat flour determination in the cooked meat products at 0.2% level (i.e. 0.375 ng of wheat protein applied to the NC strip). Since the wheat antibody (primary antiserum) has no crossreactivity with pork or beef, this technique can be used for the detection of plant proteins in the mentioned mixtures.

Flour↗

IgE binding to pepsin-digested water soluble and insoluble wheat proteins.

BACKGROUND: Positive skin prick tests to wheat are a common finding among atopic patients, but only a minor fraction of these patients show immediate clinical symptoms after wheat ingestion. The aim of this study was to evaluate the allergenicity of wheat proteins after pepsin treatments. METHODS: Six patients with positive specific IgE and/or skin prick test to soluble wheat proteins and to gluten were studied. Five of them had no symptoms after wheat ingestion and showed a negative oral challenge test. All sera were analysed with extracts obtained after pepsin hydrolysis during different time periods of water-soluble and insoluble wheat proteins by IgE immunoblotting. RESULTS: A pepsin-sensitive allergen of around 35 kDa was recognized by the five atopic negative wheat oral challenge patients in the undigested water-insoluble extract. A patient showing immediate urticaria after wheat ingestion detected a pepsin-resistant allergen with similar molecular weight. Finally, an inhalation-induced asthma patient recognized water-soluble proteins (of around 14 kDa) that could correspond to the alpha-amylase inhibitors. CONCLUSIONS: Our immunoblotting method shows better correlation with the clinical symptoms than skin prick test and CAP results. Pepsin resistance of the acetic acid-soluble wheat proteins may be responsible for the immediate symptoms of the patient who presented immediate urticaria.

Adolescent↗

IgE antibodies to omega-5 gliadin in children with wheat-induced anaphylaxis.

BACKGROUND: Wheat can cause severe immunoglobulin E (IgE)-mediated systemic reactions including anaphylaxis but knowledge on relevant wheat allergens at the molecular level is scanty. METHODS: Seven children (aged from 6 months to 13 years) experiencing from 2 to 10 anaphylactic reactions in a year after eating food-containing wheat were examined. Purified omega-5 gliadin was used as an allergen in IgE enzyme-linked immunosorbent assay (ELISA) and in skin prick testing (SPT). Wheat CAP radioallergosorbent test (RAST) and SPT were also examined. RESULTS: All seven anaphylactic children, but none of 15 control subjects had IgE antibodies to omega-5 gliadin in ELISA. Five of the six tested anaphylactic children showed positive SPT to omega-5 and crude gliadin, and all seven had positive wheat CAP RAST and SPT. One child was challenged with wheat, which caused anaphylaxis. After adherence to a wheat-free diet four children remained symptomless and three experienced one to two anaphylactic reactions. CONCLUSION: The present results show that wheat omega-5 gliadin is a major sensitizing allergen in children with wheat-induced anaphylaxis. They also suggest that omega-5 gliadin IgE ELISA could be used as a diagnostic test for this severe allergy.

Adolescent↗

[Fermented wheat germ extract in the supportive therapy of colorectal cancer].

The role of the product in the treatment of colorectal cancer is reviewed in the light of experimental and clinical results to date. The fermented wheat germ extract (code name: MSC, trade name: Avemar) registered as a dietary food for special medical purposes for cancer patients to complement the active oncotherapy, exerted a growth inhibitory effect in HCR-25 human colon carcinoma xenograft, and had a synergistic effect with 5-FU in mouse C-38 colorectal carcinoma. The product is capable of chemoprevention of colon carcinoma in F-344 rats. One of the most significant underlying mechanism is a highly cancer cell specific induction of caspase-3 mediated cleavage of PARP. In the frame of supportive therapy, fermented wheat germ extract proved to be efficient in the treatment of colorectal cancer in humans. 30 patients following radical operation were treated with standard postoperative therapy, 12 of them were given fermented wheat germ extract as additive treatment: following a 9 month long administration, no new distant metastases were detected, in contrast to 4 out 18 treated with standard therapy alone. Out of 34 patients following radical surgery and treated with chemotherapy, 17 who were given fermented wheat germ extract, achieved an improved survival rate. In the frame of a controlled multicenter open label cohort study, 170 colorectal cancer patients received anticancer therapies (chemo/radiotherapy) completed with fermented wheat germ extract in 66 of them. Results (fermented wheat germ extract vs. control): new recurrences: 3.0% vs. 17.3% (p < 0.01); new metastases: 7.6% vs. 23.1% (p < 0.01); deaths: 12.1% vs. 31.7% (p < 0.01), progression-related events in total: 16.7% vs. 42.3% (p < 0.001). Survival analysis showed significant improvements in the fermented wheat germ extract group, regarding progression-free (p = 0.0184) and overall survival probabilities (p = 0.0278). Strong predictors of survival determined by Cox's proportional hazards were UICC stage and fermented wheat germ extract treatment. Mild gastrointestinal side effects were observed in 9 cases. Supportive application of fermented wheat germ extract in colorectal cancer is highly recommended.

Adult↗

[Diagnosing allergic reactions to ingested wheat in children].

BACKGROUND: Sensitisation to wheat demonstrated by specific IgE without clinical reactions to ingested wheat is common. Clinical allergy towards wheat may cause anaphylactic or mild reactions. Until now, oral challenge tests have been necessary for diagnosing severe reactions to ingested wheat. We wanted to distinguish between severe and mild reactions by using three different IgE tests against wheat. MATERIAL AND METHODS: 29 children with elevated levels of specific IgE against wheat (f4 test) were referred to our hospital for further diagnostic procedures. We performed specific IgE tests against different wheat components (f4, f79 and f98) and oral provocation tests. RESULTS: 10 children with severe clinical reactions had high levels at all three specific IgE tests and comparable levels between the tests. The 4 children with mild reactions showed varying values of IgE but very similar levels between the three different tests. 15 children had negative provocation tests, varying levels of specific IgE, and less similar levels between the tests. INTERPRETATION: Available specific IgE tests are not useful in order to distinguish between severe, mild or no clinical reactions to ingested wheat. Oral challenge tests are still needed for correct diagnosis of severe wheat allergy.

Allergens↗

Very high CO2 reduces photosynthesis, dark respiration and yield in wheat.

Although terrestrial CO2 concentrations, [CO2] are not expected to reach 1000 micromoles mol-1 for many decades, CO2 levels in closed systems such as growth chambers and glasshouses, can easily exceed this concentration. CO2 levels in life support systems in space can exceed 10000 micromoles mol-1 (1%). Here we studied the effect of six CO2 concentrations, from ambient up to 10000 micromoles mol-1, on seed yield, growth and gas exchange of two wheat cultivars (USU-Apogee and Veery-l0). Elevating [CO2] from 350 to 1000 micromoles mol-1 increased seed yield (by 33%), vegetative biomass (by 25%) and number of heads m-2 (by 34%) of wheat plants. Elevation of [CO2] from 1000 to 10000 micromoles mol-1 decreased seed yield (by 37%), harvest index (by 14%), mass per seed (by 9%) and number of seeds per head (by 29%). This very high [CO2] had a negligible, non-significant effect on vegetative biomass, number of heads m-2 and seed mass per head. A sharp decrease in seed yield, harvest index and seeds per head occurred by elevating [CO2] from 1000 to 2600 micromoles mol-1. Further elevation of [CO2] from 2600 to 10000 micromoles mol-1 caused a further but smaller decrease. The effect of CO2 on both wheat cultivars was similar for all growth parameters. Similarly there were no differences in the response to high [CO2] between wheat grown hydroponically in growth chambers under fluorescent lights and those grown in soilless media in a glasshouse under sunlight and high pressure sodium lamps. There was no correlation between high [CO2] and ethylene production by flag leaves or by wheat heads. Therefore, the reduction in seed set in wheat plants is not mediated by ethylene. The photosynthetic rate of whole wheat plants was 8% lower and dark respiration of the wheat heads 25% lower when exposed to 2600 micromoles mol-1 CO2 compared to ambient [CO2]. It is concluded that the reduction in the seed set can be mainly explained by the reduction in the dark respiration in wheat heads, when most of the respiration is functional and is needed for seed development.

Biomass↗

Homology in the region containing a tRNA(Trp) gene and a (complete or partial) tRNA(Pro) gene in wheat mitochondrial and chloroplast genomes.

We have used bean mitochondrial (mt) and chloroplast (cp) tRNA(Trp) as probes to locate the corresponding genes on the mt and cp genomes of wheat and we have determined the nucleotide sequences of the wheat mt and cp tRNA(Trp) genes and of the flanking regions. Sequence comparisons show that the wheat mt and cp tRNA(Trp) genes are 97% homologous. On the wheat cp DNA, a tRNA(UGGPro) gene was found 139 bp upstream of the cp tRNA(Trp) gene. On the wheat mt DNA, a sequence of 23 nucleotides completely homologous with the 3' end of this cp tRNA(Pro) gene was found 136 bp upstream of the mt tRNA(Trp) gene, but there is only 38% homology between cp and mt wheat genomes in the intergenic regions. The overall organization of this region in the chloroplast genome (a tRNA(Trp) gene separated by about 140 bp from a tRNA(Pro) gene) is also found in the mitochondrial genome, suggesting that this mitochondrial fragment might have originated from a chloroplast DNA insertion. A comparison of the genes and of the intergenic regions located between the tRNA(Trp) gene and the tRNA(Pro) (or partial tRNA(Pro)) gene shows that there is an almost complete conservation of these sequences in the mitochondrial DNA of wheat and maize, whereas wheat mt and cp intergenic regions show more sequence divergence. Wheat mt tRNA(Trp) gene is encoded by the main mt genome (accounted for by the master chromosome) but, in the case of maize mitochondria, this gene was found to be encoded by the 2.3 kb linear plasmid, indicating that this plasmid is not dispensable in maize mitochondria.

Base Sequence↗

Transferability of SSR markers among wheat, rye, and triticale.

Simple sequence repeat (SSR) markers are a valuable tool for many purposes, such as mapping, fingerprinting, and breeding. However, they are only available in some economically important crops because of the high cost and labor intensity involved in their development. Comparative mapping reveals a high degree of colinearity between closely related species, which allows the exchange of markers between them. Our objective was to examine the transferability of SSR markers among wheat ( Triticum aestivum L.), rye ( Secale cereale L.), and triticale (X Triticosecale Wittmack). One hundred forty-eight wheat and 28 rye SSR markers were used to amplify genomic DNA extracted from five lines each of wheat, rye, and triticale. Transferability of wheat SSR markers to rye was 17%, whereas 25% of rye markers were amplifiable in wheat. In triticale, 58% and 39% transferability was achieved for wheat and rye markers, respectively. Wheat markers gave an average of 2.6, 2.7, and 2.4 polymorphic bands in wheat, rye, and triticale, respectively, while rye markers gave an average of 2.0 in rye and none in wheat and triticale. These transferable markers can now be exploited for further genetic and breeding studies in these species.

Edible Grain↗

Complementation of the pina (null) allele with the wild type Pina sequence restores a soft phenotype in transgenic wheat.

The tightly linked puroindoline genes, Pina and Pinb, control grain texture in wheat, with wild type forms of both giving soft, and a sequence alteration affecting protein expression or function in either giving rise to hard wheat. Previous experiments have shown that addition of wild type Pina in the presence of mutated Pinb gave intermediate grain texture but addition of wild type Pinb gave soft grain. This raises questions as to whether Pina may be less functional than Pinb. Our goal here was to develop and characterize wheat lines expressing the wild type Pina-D1a sequence in hard wheat with the null mutation (Pina-D1b) for Pina. Three transgenic lines plus Bobwhite were evaluated in two environments. Grain texture, grain protein, and kernel weight were determined for the transgenic lines and Bobwhite. The three transgenic lines had soft phenotype, and none of the transgenic lines differed from Bobwhite for grain protein or kernel weight. The soft phenotype was accompanied by increases in Pina transcript accumulation. Total Triton X-114 extractable PINA and PINB increased from 2.5 to 5.5 times those from a soft wheat reference sample, and friabilin, PINA and PINB bound to starch, increased from 3.8 to 7.8 times those of the soft wheat reference. Bobwhite showed no starch bound PINA, but transgenic lines had levels from 5.3 to 13.7 times those of the soft wheat reference sample. Starch bound PINB in transgenic lines also increased from 0.9 to 2.5 times that for the soft wheat reference sample. The transgenic expression of wild type Pina sequence in the Pina null genotype gave soft grain with the characteristics of soft wheat including increased starch bound friabilin. The results support the hypothesis that both wild type Pin genes need to be present for friabilin formation and soft grain.

Alleles↗

Detailed comparison between the wheat chromosome group 7 short arms and the rice chromosome arms 6S and 8L with special reference to genes involved in starch biosynthesis.

Rice bacterial artificial chromosome (BAC) clones have been identified that contain sequences orthologous to each EST localized to wheat chromosome 7AS deletion stocks by Southern blot hybridization. This information has been used to relate the DNA sequence included in each wheat deletion stock to a complement of rice BACs. A virtual contig was used that covered 90 cM (21 Mb) of DNA sequence (with a gap for the 6S/8L junction). Comparison of the positions of orthologous genes on the rice virtual contig and on wheat chromosome 7AS showed that there was an unexpectedly low level of synteny (31.4%) and a high level of chromosome rearrangements (68.6%). The non-syntenous loci were of two classes: wheat and rice genes found at different locations in the genome (32.6%), and ESTs in wheat not present in rice (36.0%). Four starch synthetic genes, GBSSI, SSI, SSIIa and DBEI, were located at similar positions on wheat chromosome 7AS and the virtual rice contig covering wheat chromosome 7AS. A preliminary comparison between the short arms of chromosome 7A and 7D in wheat showed that both chromosomes had a similar level of sequence synteny with rice. Therefore, there appears to be considerable variation in gene order between wheat chromosome 7S and rice chromosome 6S and 8L.

Chromosomes, Artificial, Bacterial↗

Comparative pathogenicity of Fusarium graminearum isolates from China revealed by wheat coleoptile and floret inoculations.

Fusarium head blight (FHB) or scab caused by Fusarium species is an economically important disease on small grain cereal crops worldwide. Accurate assessments of the pathogenicity of fungal isolates is a key obstacle toward a better understanding of the Fusarium-wheat scab system. In this study, a new laboratory method for inoculation of wheat coleoptiles was developed, which consists of cutting off the coleoptile apex, covering the cut apex with a piece of filter paper soaked in conidial suspension, and measuring the lengths of brown lesions 7 days post inoculation. After coleoptile inoculation, distinct brown lesions in the diseased stems were observed, in which the presence of the fungus was verified by PCR amplification with F. graminearum Schwable-specific primers. Coleoptile inoculation of six wheat varieties indicated that a highly susceptible wheat variety was more suitable as a differentiating host for the pathogenicity assay. Analysis of the coleoptiles inoculated with a set of 58 different isolates of F. graminearum showed a significant difference in the lengths of the lesions, forming the basis by which pathogenicity of the isolates was assessed. Field inoculation of florets of three wheat varieties over 2 years revealed significant differences in pathogenicity among the 58 isolates, and that the highly resistant and highly susceptible wheat varieties were more appropriate and stable for pathogenicity assessment in field trials. Comparative analyses of eight inoculation experiments of wheat with 58 F. graminearum isolates showed significant direct linear correlations (P<0.001) between coleoptile and floret inoculations. These results indicate that the wheat coleoptile inoculation is a simple, rapid and reliable method for pathogenicity studies of F. graminearum in wheat.

China↗

Comparative sequence analysis of the phytochrome C gene and its upstream region in allohexaploid wheat reveals new data on the evolution of its three constituent genomes.

Bread wheat is an allohexaploid with genome composition AABBDD. Phytochrome C is a gene involved in photomorphogenesis that has been used extensively for phylogenetic analyses. In wheat, the PhyC genes are single copy in each of the three homoeologous genomes and map to orthologous positions on the long arms of the group 5 chromosomes. Comparative sequence analysis of the three homoeologous copies of the wheat PhyC gene and of some 5 kb of upstream region has demonstrated a high level of conservation of PhyC, but frequent interruption of the upstream regions by the insertion of retroelements and other repeats. One of the repeats in the region under investigation appeared to have inserted before the divergence of the diploid wheat genomes, but was degraded to the extent that similarity between the A and D copies could only be observed at the amino acid level. Evidence was found for the differential presence of a foldback element and a miniature inverted-repeat transposable element (MITE) 5' to PhyC in different wheat cultivars. The latter may represent the first example of an active MITE family in the wheat genome. Several conserved non-coding sequences were also identified that may represent functional regulatory elements. The level of sequence divergence (Ks) between the three wheat PhyC homoeologs suggests that the divergence of the diploid wheat ancestors occurred some 6.9 Mya, which is considerably earlier than the previously estimated 2.5-4.5 Mya. Ka/Ks ratios were <0.15 indicating that all three homoeologs are under purifying selection and presumably represent functional PhyC genes. RT-PCR confirmed expression of the A, B and D copies. The discrepancy in evolutionary age of the wheat genomes estimated using sequences from different parts of the genome may reflect a mosaic origin of some of the Triticeae genomes.

5' Flanking Region↗

Changes of some anti-oxidative physiological indices under soil water deficits among 10 wheat (Triticum aestivum L.) genotypes at tillering stage.

Drought is one of the major ecological factors limiting crop production and food quality globally, especially in the arid and semi-arid areas of the world. Wheat is the staple food for more than 35% of world population and wheat cultivation is mainly restricted to such zones with scarcity of water, so wheat anti-drought physiology study is of importance to wheat production, food safety and quality and biotechnological breeding for the sake of coping with abiotic and biotic conditions. The current study is to investigate changes of anti-oxidative physiological indices of 10 wheat genotypes at tillering stage. The main results and conclusion of tillering stage in terms of activities of POD, SOD, CAT and MDA content as followed: (1) 10 wheat genotypes can be generally grouped into three kinds (A-C, respectively) according to their changing trend of the measured indices; (2) A group performed better drought resistance under the condition of treatment level 1 (appropriate level), whose activities of anti-oxidative enzymes (POD, SOD, CAT) were higher and MDA lower; (3) B group exhibited stronger anti-drought under treatment level 2 (light-stress level), whose activities of anti-oxidative enzymes were higher and MDA lower; (4) C group expressed anti-drought to some extent under treatment level 3 (serious-stress), whose activities of anti-oxidative enzymes were stronger, MDA lower; (5) these results demonstrated that different wheat genotypes have different physiological mechanisms to adapt themselves to changing drought stress, whose molecular basis is discrete gene expression profiling (transcriptom). The study in this respect is the key to wheat anti-drought and biological-saving water in worldwide arid and semi-arid areas; (6) POD, SOD, and CAT activities and MDA content of different wheat genotypes had quite different changing trend at different stages and under different soil water stress conditions, which was linked with their origin of cultivation and individual soil water threshold, which will provide better reference to selecting proper plant species for eco-environmental construction and crops for sustainable agriculture in arid and semi-arid areas.

Antioxidants↗

Analysis of recombination and gene distribution in the 2L1.0 region of wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.).

Both wheat and barley belong to tribe Triticeae and are closely related. High-density detailed comparison of physical and genetic linkage maps revealed that wheat genes are present in physically small gene-rich regions (GRRs). One of the largest GRRs is located around fraction length 1.0 of the long arm of wheat homoeologous group 2 chromosomes termed the "2L1.0 region." The main objective of this study was to analyze the structural and functional organization of the 2L1.0 region in barley in comparison to wheat. Using the 29 physically mapped RFLP markers for the region, wheat and barley consensus genetic linkage maps of the 2L1.0 region were generated by combining information from 18 wheat and 7 barley genetic linkage maps. Comparative analysis using these consensus maps and other available wheat and barley mapping resources identified 227 DNA markers and ESTs for the region. The region accounted for 58% of the genes and 68% of the arm's recombination in wheat. However, the corresponding region in barley accounted for about 42% of the genes and 81% of the recombination. The kb/cM ratio for the region was 122 in barley compared to 244 in wheat. Distribution of genes and recombination varied between the two species even though the gene order and density were similar.

Chromosome Mapping↗

Wheat starch intolerance in patients with celiac disease.

OBJECTIVE: Evaluate in patients with celiac disease the tolerance of prolonged consumption of small amounts of gliadin contained in products containing wheat starch. DESIGN: Open 1-year trial of the addition of wheat starch to a gluten-free diet in a cohort of adult patients with biopsy-proven celiac disease who had never consumed wheat starch. The control group consisted of patients with celiac disease who tolerated wheat starch. SUBJECTS: Seventeen patients with celiac disease and 14 control patients, all diagnosed according to criteria of the European Society of Pediatric Gastroenterology and Nutrition, were recruited from the Canadian Celiac Association and the Quebec Celiac Foundation. SETTING: The study was conducted in the outpatient clinic of the Gastroenterology and Nutrition Service of Ste Justine Hospital, Montreal, Quebec, Canada. INTERVENTIONS: Patients were asked to consume four to six portions daily of a wheat starch-containing product, mainly bread, for up to 1 year. MAIN OUTCOME MEASURES: The gliadin content of the wheat starch product used in this trial was quantified by enzyme-linked immunosorbent assay. Patient outcome measures included symptoms, nutritional parameters (anthropometric data, complete blood count, serum folate and iron levels), and immunologic parameters (antigliadin antibody and antiendomysium antibody titers). RESULTS: A quantifiable amount of immunoreactive gliadin (0.75 mg/100 g) was found in the wheat starch. The majority of the patients with celiac disease (11 of 17) who had never consumed wheat starch previously developed symptoms, which resolved within weeks of discontinuing the product. Relapse of skin lesions was seen in two of three patients with coexisting dermatitis herpetiformis. No weight loss or biochemical changes were observed. Despite the presence of symptoms, antigliadin antibody and antiendomysium antibody determinations were not useful to detect the clinical intolerance. APPLICATIONS: The innocuousness of the long-term ingestion of "gluten-free" products containing wheat starch is still unproven, and prolonged use of such products by patients with celiac disease cannot be recommended.

Adolescent↗

A novel wheat gliadin as a cause of exercise-induced anaphylaxis.

BACKGROUND: Food-dependent, exercise-induced anaphylaxis is a severe form of allergy; the reaction is caused by ingestion of a specific food before exercise. This disorder often escapes diagnosis because neither the ingested food nor the exercise alone induces the symptoms. OBJECTIVE: The aim of the study was to characterize the allergens involved in wheat-dependent, exercise-induced anaphylaxis and to describe the clinical outcome in a series of 18 adult patients. METHODS: All 18 patients had experienced recurrent episodes of generalized urticaria during exercise, 17 patients in association with collapse and 15 patients with an anaphylactic reaction. The symptoms appeared only when the patients had eaten food containing wheat before exercise. Wheat allergens were detected by immunoblotting, purified by gel filtration and reversed-phase chromatography, and subjected to N-terminal sequencing. The IgE-binding ability of the purified proteins was studied by ELISA, and their in vivo reactivity was studied by skin prick testing. RESULTS: IgE antibodies from pooled patient sera were bound to 65-kd and 40-kd wheat proteins in immunoblotting. The 65-kd allergen was a previously undescribed wheat protein, showing 61% sequence identity to gamma-gliadin, whereas the 40-kd allergen had 100% identity to alpha-gliadin. In ELISA, all 18 patients showed elevated IgE levels to the novel gamma-like gliadin, and 13 of the patients showed elevated IgE levels to the alpha-gliadin. None of the 54 control subjects with wheat allergy, urticaria, or coeliac disease had IgE antibodies to the gamma-like gliadin. The in vivo reactivity of the gamma-like gliadin was verified by positive skin prick test responses in all of the 15 patients who were tested. During the follow-up on a gluten-free or wheat-free diet, 3 patients experienced reactions after having unknowingly eaten wheat before exercise, but all the other patients who were adhering to the diet remained symptom-free. CONCLUSION: This study shows that wheat is a frequent cause of food-dependent, exercise-induced anaphylaxis and suggests that the major allergen is a previously undescribed gamma-like gliadin. For screening of this life-threatening allergy, we recommend skin prick testing with crude gliadin and we recommend a gluten-free diet for treatment.

Adolescent↗

Fast omega-gliadin is a major allergen in wheat-dependent exercise-induced anaphylaxis.

BACKGROUND: Wheat-dependent exercise-induced anaphylaxis is an anaphylaxy induced by physical exercise after ingestion of wheat. An immediate-type hypersensitivity to water/salt-insoluble fraction of wheat proteins (gluten) has been considered to underlie in this disease. OBJECTIVE: The aim of the study is to determine the major allergen in Japanese patients with wheat-dependent exercise-induced anaphylaxis by using a panel of purified wheat gliadins and glutenins. METHODS: Water/salt-insoluble wheat proteins, alpha-gliadin, beta-gliadin, gamma-gliadin, fast omega-gliadin, slow omega-gliadin, high molecular weight glutenin and low molecular weight glutenin, were purified, and five patients with wheat-dependent exercise-induced anaphylaxis, whose diagnose had been determined by positive-challenge test, were evaluated for skin prick test, dot-blotting test and CAP-RAST inhibition test by using these purified wheat proteins. RESULTS: The fast omega-gliadin was the most potent allergen among these water/salt-insoluble proteins when evaluated by skin prick test and dot-blotting test. Fast and slow omega-gliadin, and gamma-gliadin caused dose-dependent inhibition of the serum IgE-binding to solid-phase gluten in the patients. The incubation with fast omega-gliadin of the patient's serum caused dose-dependent inhibition in the IgE-binding to gamma-gliadin as well as slow omega-gliadin, indicating a cross-reactivity of these proteins in IgE-binding. CONCLUSION: We concluded that fast omega-gliadin is a major allergen among these water/salt-insoluble proteins for wheat-dependent exercise-induced anaphylaxis in Japanese patients, and IgE against fast omega-gliadin cross-reacts to gamma-gliadin and slow omega-gliadin.

Adolescent↗