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PubMed2005

Introgression of wheat DNA markers from A, B and D genomes in early generation progeny of Aegilops cylindrica Host x Triticum aestivum L. hybrids.

Introgression from allohexaploid wheat (Triticum aestivum L., AABBDD) to allotetraploid jointed goatgrass (Aegilops cylindrica Host, CCDD) can take place in areas where the two species grow in sympatry and hybridize. Wheat and Ae. cylindrica share the D genome, issued from the common diploid ancestor Aegilops tauschii Coss. It has been proposed that the A and B genome of bread wheat are secure places to insert transgenes to avoid their introgression into Ae. cylindrica because during meiosis in pentaploid hybrids, A and B genome chromosomes form univalents and tend to be eliminated whereas recombination takes place only in D genome chromosomes. Wheat random amplified polymorphic DNA (RAPD) fragments, detected in intergeneric hybrids and introgressed to the first backcross generation with Ae. cylindrica as the recurrent parent and having a euploid Ae. cylindrica chromosome number or one supernumerary chromosome, were assigned to wheat chromosomes using Chinese Spring nulli-tetrasomic wheat lines. Introgressed fragments were not limited to the D genome of wheat, but specific fragments of A and B genomes were also present in the BC1. Their presence indicates that DNA from any of the wheat genomes can introgress into Ae. cylindrica. Successfully located RAPD fragments were then converted into highly specific and easy-to-use sequence characterised amplified regions (SCARs) through sequencing and primer design. Subsequently these markers were used to characterise introgression of wheat DNA into a BC1S1 family. Implications for risk assessment of genetically modified wheat are discussed.

Base Sequence↗
PubMed2001

The high level of aluminum resistance in signalgrass is not associated with known mechanisms of external aluminum detoxification in root apices.

Al resistance of signalgrass (Brachiaria decumbens Stapf cv Basilisk), a widely sown tropical forage grass, is outstanding compared with the closely related ruzigrass (Brachiaria ruziziensis Germain and Evrard cv Common) and Al-resistant genotypes of graminaceous crops such as wheat, triticale, and maize. Secretion of organic acids and phosphate by root apices and alkalinization of the apical rhizosphere are commonly believed to be important mechanisms of Al resistance. However, root apices of signalgrass secreted only moderately larger quantities of organic acids than did those of ruzigrass, and efflux from signalgrass apices was three to 30 times smaller than from apices of Al-resistant genotypes of buckwheat, maize, and wheat (all much more sensitive to Al than signalgrass). In the presence, but not absence, of Al, root apices of signalgrass alkalinized the rhizosphere more than did those of ruzigrass. The latter was associated with a shortening of the alkalinizing zone in Al-intoxicated apices of ruzigrass, indicating that differences in alkalinizing power were a consequence, not a cause of, differential Al resistance. These data indicate that the main mechanism of Al resistance in signalgrass does not involve external detoxification of Al. Therefore, highly effective resistance mechanisms based on different physiological strategies appear to operate in this species.

Aluminum↗
PubMed1976

Flour allergy in bakers. I. Identification of allergenic fractions in flour and comparison of diagnostic methods.

Extract of wheat flour obtained by extraction, centrifugation and dialysis was immunochemically characterized by quantitative immunoelectrophoresis using rabbit antibodies. The analysis revealed wheat flour to be composed of 40 antigens, some of which were immunologically partially identical with antigens of rye flour and of common grass pollen. Furthermore, antigens of the gliadin fraction of wheat flour were identified. 25 bakers with allergic complaints working in and around Copenhagen were clinically tested with wheat flour and related extracts. Among 13 bakers with respiratory complaints (asthma and/or rhinitis), 11 showed positive reactions to wheat flour extract when tested in histamine release from basophil leukocytes radioallergosorbent test and skin test, whereas further 2 were positive in the basophil test only. The ability of the IgE of individual sera to adsorb to the individual antigens of wheat flour was examined by means of crossed radioimmunoelectrophoresis. On the basis of these results, individual allergenic components of wheat flour were identified, three of these with comparatively high affinity and frequency.

Antigens↗
PubMed1995

Impact of Field Release of Genetically Modified Pseudomonas fluorescens on Indigenous Microbial Populations of Wheat.

In a field release experiment, an isolate of Pseudomonas fluorescens, which was chromosomally modified with two reporter gene cassettes (lacZY and Kan(supr)-xylE), was applied to spring wheat as a seed coating and subsequently as a foliar spray. The wild-type strain was isolated from the phylloplane of sugar beet but was found to be a common colonizer of both the rizosphere and phylloplane of wheat as well. The impact on the indigenous microbial populations resulting from release of this genetically modified microorganism (GMM) was compared with the impact of the unmodified, wild-type strain and a nontreated control until 1 month after harvest of the crop. The release of the P. fluorescens GMM and the unmodified, wild-type strain resulted in significant but transient perturbations of some of the culturable components of the indigenous microbial communities that inhabited the rhizosphere and phylloplane of wheat, but no significant perturbations of the indigenous culturable microbial populations in nonrhizosphere soil were found. Fast-growing organisms that did not produce resting structures (for example, fluorescent pseudomonads and yeasts) seemed to be most sensitive to perturbation. In terms of hazard and risk to the environment, the observed microbial perturbations that resulted from this GMM release may be considered minor for several reasons. First, the recombinant P. fluorescens strain caused changes that were, in general, not significantly different from those caused by the unmodified wild-type strain; second, perturbations resulting from bacterial inoculations were mainly small; and third, the release of bacteria had no obvious effects on plant growth and plant health.

Journal Article↗
PubMed1988

Detection of fungi in tissue sections by lectin histochemistry.

A battery of 21 fluoresceinated lectins was tested on sections of paraffin-embedded human tissues to determine which lectin could be used in the microscopic diagnosis of fungal diseases. Three lectins, from Canavalia ensiformis (concanavalin), Triticum vulgare (wheat germ A), and Phytolacca americana (pokeweed [PWM]), were found to react with fungal pathogens commonly encountered in nosocomial infections. Best results were obtained with PWM and succinylated wheat germ A, which, due to their weak reactivity with normal tissues, reacted almost selectively with fungi.

Fluorescein-5-isothiocyanate↗
PubMed2004

Wheat cultivar-specific selection of 2,4-diacetylphloroglucinol-producing fluorescent Pseudomonas species from resident soil populations.

An emerging body of evidence indicates a role for plant genotype as a determinant of the species and genetic composition of the saprophytic microbial community resident to the rhizosphere. In this study, experiments were conducted to determine the capacity of five different wheat cultivars to enhance resident populations and support introduced strains of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent pseudomonads, a group of bacteria known to provide biological control of several soilborne diseases. When soils were cropped with three successive 28-day growth cycles of wheat, the 2,4-DAPG-producing strains were consistently recovered from the rhizosphere of the cultivar Lewjain, and commonly were present at populations higher than those recovered from other wheat cultivars. Based on restriction fragment length polymorphism and sequence analyses of phlD, a key gene involved in 2,4-DAPG production, two previously undefined phlD+ genotypes, referred to as genotypes PfZ and PfY, were discovered. Wheat cultivar Lewjain was the primary source of genotype PfY while cultivar Penawawa yielded the majority of genotype PfZ. Based on 16S rDNA sequence analysis, both new phlD genotypes were classified as P. fluorescens. Comparison of the rhizosphere competence of 2,4-DAPG-producing P. fluorescens Q2-87 (genotype B) and P. fluorescens LR3-A28 (genotype PfY) showed that both strains persisted at similar populations in the rhizosphere of all cultivars tested over a 30 day period when introduced as a seed inoculant. However, when strain LR3-A28 was applied as a soil inoculant, this strain was recovered at higher populations from the rhizosphere of wheat cultivar Lewjain than from the rhizospheres of two other cultivars. No cultivar effects were shown for strain Q2-87. Collectively, these results add further to evidence indicating a degree of specificity in interactions between plant cultivars and specific members of the saprophytic microbial community. Furthermore, as 2,4-DAPG-producing fluorescent Pseudomonas spp. have a central role in the spontaneous reduction in severity of take-all disease of wheat in response to continuous wheat monoculture, we postulate that the use of specific cultivars, such as Lewjain, which possess a superior capacity to enhance resident soil populations of these bacteria may have potential to reduce the length of the monoculture period required to induce natural suppressiveness of soils toward this disease.

Bacterial Proteins↗
PubMed1987

Selective inhibition of antibody-dependent allergic autocytotoxicity by OM-89, in patients with bronchial asthma and urticaria having food allergy toward wheat.

Twenty-five patients with bronchial asthma and urticaria expressing hypersensitivity to ingested wheat were examined and compared with fourteen controls using the allergic autocytotoxicity assay (ACT). To establish a possible interaction between wheat antigen and antibody with OM-89 (immunostimulating fraction extracted from E. coli), peripheral WBC of the above individuals were tested in vitro. Preincubation with OM-89 and subsequent exposure toward wheat antigen in the direct ACT assay and antibody-dependent ACT assay using specific wheat antibody were performed. A separate study of spontaneous ACT was conducted in which unprimed white blood cells were pre-incubated with OM-89 alone. Our results show that OM-89 exerted its most potent inhibitory activity in the antibody-dependent ACT assay when the wheat antigen-antibody test system was used. Only borderline inhibition of the direct ACT response was observed, while OM-89 did not exert any effect on spontaneous ACT. This observation may be of importance in future clinical trials since patients sensitive to wheat with high antibody levels toward this common ingestant antigen may benefit from oral therapy with OM-89.

Adjuvants, Immunologic↗
PubMed2005

Food allergen sensitization in inner-city children with asthma.

BACKGROUND: Asthma continues to be an increasing cause of morbidity in the pediatric population, and studies have shown an association between food sensitivity and asthma. OBJECTIVE: We investigated the degree of food allergen sensitization in inner-city patients with asthma. METHODS: Five hundred four random serum samples from the National Cooperative Inner City Asthma Study were evaluated for specific IgE (UniCap) to 6 common food allergens (egg, milk, soy, peanut, wheat, and fish). Statistical analyses were performed to determine food sensitization prevalence and its association with asthma morbidity. RESULTS: Forty-five percent of patients had evidence of sensitization (food-specific IgE > or = 0.35 kU/L) to at least 1 food. Nineteen percent had IgE levels at > or = 50% positive predictive value for clinical reactivity to at least 1 food, with 4% of patients having levels > 95% positive predictive value for food allergy. Children sensitized to foods had higher rates of asthma hospitalization (P < .01) and required more steroid medications (P = .025). Sensitization to foods also correlated with sensitization to more indoor and outdoor aeroallergens (P < .001). CONCLUSION: Food allergen sensitization is highly prevalent in the inner-city population with asthma, and it is associated with increased asthma healthcare and medication use. Therefore, food allergen sensitivity may be a marker for increased asthma severity.

Animals↗
PubMed2003

Molecular identification of wheat endoxylanase inhibitor TAXI-I1, member of a new class of plant proteins.

Triticum aestivum endoxylanase inhibitors (TAXIs) are wheat proteins that inhibit family 11 endoxylanases commonly used in different (bio)technological processes. Here, we report on the identification of the TAXI-I gene which encodes a mature protein of 381 amino acids with a calculated molecular mass of 38.8 kDa. When expressed in Escherichia coli, the recombinant protein had the specificity and inhibitory activity of natural TAXI-I, providing conclusive evidence that the isolated gene encodes an endoxylanase inhibitor. Bioinformatical analysis indicated that no conserved domains nor motifs common to other known proteins are present. Sequence analysis revealed similarity with a glycoprotein of carrot and with gene families in Arabidopsis thaliana and rice, all with unknown functions. Our data indicate that TAXI-I belongs to a newly identified class of plant proteins for which a molecular function as glycoside hydrolase inhibitor can now be suggested.

Amino Acid Sequence↗
PubMed1989

Fusarium moniliforme Sheld. A fungus producing a broad spectrum of bioactive metabolites.

Fusarium moniliforme Sheldon (perfect stage Gibberella fujikuroi (SAW. Wollenweber), an ubiquitous fungus belonging to the section Liseola (Booth), is one of the widespread, phytopathogenic fungi. This field fungus common on many crops, including rice, oats, wheat, maize, barley and soybeans was well known because of the ability to produce gibberellins in high quantities. Today beside of the very important gibberellic acid (GA3) other gibberellins such as GA4 and GA7 are of significance for agriculture and horticulture. Moreover, this fungus produces further biologically active metabolites, e.g. pigments, mycotoxins, phytotoxins, oestrogens and extracellular enzymes. Fusarium moniliforme strains are able to produce 2 pigment groups - the karotenoids and the bikaverins, the last of which has antibiotical activity against Leishmania brasiliensis. The biosynthesis of some mycotoxic substances, such as moniliformin, fusaric acid, fusarin C and fusariocin C, is described. From enzymes the milk-clotting rennin, cellulolytic and amylolytic enzymes, pectinases and phenol-degrading enzymes are of economical interest. Therefore, Fusarium moniliforme is a source of different bioactive metabolites. The production is dependent on the conditions and strain specifity.

Chemical Phenomena↗
PubMed1999

Food allergy: when and how to perform oral food challenges.

In many situations, the diagnosis of food allergy rests simply upon a history of an acute onset of typical symptoms, such as hives and wheezing, following the isolated ingestion of a suspected food, with confirmatory laboratory studies of positive prick skin tests or Radioallergosorbent tests. However, the diagnosis is more complicated when multiple foods are implicated or when chronic diseases, such as asthma or atopic dermatitis, are evaluated. The diagnosis of food allergy and identification of the particular foods responsible is also more difficult when reactions are not mediated by IgE antibody, as is the case with a number of gastrointestinal food allergies. In these latter circumstances, well-devised elimination diets followed by physician-supervised oral food challenges are critical in the identification and proper treatment of these disorders. Because childhood food allergies to common allergenic foods such as milk, egg, wheat and soy are usually outgrown, oral food challenges are also an integral part of the long-term management of these children.

Antibody Specificity↗
PubMed2002

A cell-free protein synthesis system for high-throughput proteomics.

We report a cell-free system for the high-throughput synthesis and screening of gene products. The system, based on the eukaryotic translation apparatus of wheat seeds, has significant advantages over other commonly used cell-free expression systems. To maximize the yield and throughput of the system, we optimized the mRNA UTRs, designed an expression vector for large-scale protein production, and developed a new strategy to construct PCR-generated DNAs for high-throughput production of many proteins in parallel. The resulting system achieves high-yield expression and can maintain productive translation for 14 days. Additionally, in the integration of a PCR-directed system for template creation, at least 50 genes can be translated in parallel, yielding between 0.1 and 2.3 mg of protein by one person within 2 days. Assessment of correct protein folding by the products of this high-throughput protein-expression system were performed by enzymatic assays of kinases and by NMR spectroscopic analysis. The cell-free system, reported here, bypasses many of the time-consuming cloning steps of conventional expression systems and lends itself to a robotic automation for the high-throughput expression of proteins.

Cell-Free System↗
PubMed2002

Cofortification of iron-fortified flour with zinc sulfate, but not zinc oxide, decreases iron absorption in Indonesian children.

BACKGROUND: Iron deficiency is a major nutritional concern in developing countries, and food fortification is a common strategy to treat it. In Indonesia wheat flour is fortified with 60 mg Fe/kg, but because of increasing concerns about marginal zinc status in at-risk populations, consideration is being given to cofortifying flour with zinc. However, little is known about the effect of zinc fortification of flour on iron bioavailability or about the optimum form of zinc supplementation. OBJECTIVE: We measured iron and zinc bioavailability from wheat-flour dumplings containing 25 g flour fortified with 60 mg Fe/kg, either alone or with 60 mg Zn/kg as zinc oxide or as zinc sulfate. DESIGN: Ninety children aged 4-8 y were recruited and assigned randomly to the 3 groups; 86 completed the study. Iron and zinc absorption were measured with established stable-isotope methods. RESULTS: Iron absorption from the flour fortified with iron only was good (15.9 +/- 6.8%), but when corrections were made for hemoglobin concentrations, it was significantly lower from the flour cofortified with zinc sulfate (11.5 +/- 4.9%; P < 0.05) but not from the flour cofortified with zinc oxide (14.0 +/- 8.9%). Zinc absorption was not significantly different between the zinc oxide and zinc sulfate cofortified flours (24.1 +/- 8.2% compared with 23.7 +/- 11.2%; P = 0.87). CONCLUSIONS: Iron and zinc appear to be highly bioavailable from foods made from fortified flour, but zinc sulfate cofortification may have a detrimental effect on iron absorption.

Biological Availability↗
PubMed1986

epsilon-N-trimethyllysine availability regulates the rate of carnitine biosynthesis in the growing rat.

Rates of carnitine biosynthesis in mammals depend on the availability of substrates and the activity of enzymes subserving the pathway. This study was undertaken to test the hypothesis that the availability of epsilon-N-trimethyllysine is rate-limiting for synthesis of carnitine in the growing rat and to evaluate diet as a source of this precursor for carnitine biosynthesis. Rats apparently absorbed greater than 90% of a tracer dose of [methyl-3H]epsilon-N-trimethyllysine, and approximately 30% of that was incorporated into tissues as [3H]carnitine. Rats given oral supplements of epsilon-N-trimethyllysine (0.5-20 mg/d), but no dietary carnitine, excreted more carnitine than control animals receiving no dietary epsilon-N-trimethyllysine or carnitine. Rates of carnitine excretion increased in a dose-dependent manner. Tissue and serum levels of carnitine also increased with dietary epsilon-N-trimethyllysine supplementation. There was no evidence that the capacity for carnitine biosynthesis was saturated even at the highest level of oral epsilon-N-trimethyllysine supplementation. Common dietary proteins (casein, soy protein and wheat gluten) were found to be poor sources of epsilon-N-trimethyllysine for carnitine biosynthesis. The results of this study indicate that the availability of epsilon-N-trimethyllysine limits the rate of carnitine biosynthesis in the growing rat.

Administration, Oral↗
PubMed2003

Dietary animal and plant protein and human bone health: a whole foods approach.

Urinary calcium excretion is strongly related to net renal acid excretion. The catabolism of dietary protein generates ammonium ion and sulfates from sulfur-containing amino acids. Bone citrate and carbonate are mobilized to neutralize these acids, so urinary calcium increases when dietary protein increases. Common plant proteins such as soy, corn, wheat and rice have similar total S per g of protein as eggs, milk and muscle from meat, poultry and fish. Therefore increasing intake of purified proteins from either animal or plant sources similarly increases urinary calcium. The effects of a protein on urinary calcium and bone metabolism are modified by other nutrients found in that protein food source. For example, the high amount of calcium in milk compensates for urinary calcium losses generated by milk protein. Similarly, the high potassium levels of plant protein foods, such as legumes and grains, will decrease urinary calcium. The hypocalciuric effect of the high phosphate associated with the amino acids of meat at least partially offsets the hypercalciuric effect of the protein. Other food and dietary constituents such as vitamin D, isoflavones in soy, caffeine and added salt also have effects on bone health. Many of these other components are considered in the potential renal acid load of a food or diet, which predicts its effect on urinary acid and thus calcium. "Excess" dietary protein from either animal or plant proteins may be detrimental to bone health, but its effect will be modified by other nutrients in the food and total diet.

Adult↗
PubMed2004

Update on threshold doses of food allergens: implications for patients and the food industry.

PURPOSE OF REVIEW: The purpose of this review is to bring the reader up to date on the importance of assessing a food's lowest observed adverse-effect level (LOAEL) with two aims. Firstly, to help industry choose tests with a level of sensitivity capable of detecting food allergens hidden in industrial products. Secondly, to specify protective measures for highly allergic individuals in order to prevent recurrent severe anaphylaxis. The review also seeks to highlight the present issues and unsolved questions. RECENT FINDINGS: Thanks to standardized oral-provocation tests (double-blind placebo-controlled food challenges), LOAELs have been identified for many IgE-dependent food allergies. Most studies concern the pediatric population. Data is available for milk, egg, peanut, wheat flour, and sesame. The LOAELs are commonly in the range of 1-2 mg of natural foods, representing a few hundred micrograms of protein. These minimal reactive doses characterize about 1% of people allergic to milk, egg, or peanut. The level at which no observed adverse effect is seen might be a few tens of micrograms of protein for peanut. At the present time, allergy to oil seems to be restricted to unrefined cold-pressed oils. SUMMARY: Concerning IgE-dependent food allergies, the threshold dose inducing symptoms is now known to vary a great deal according to the individual. A reactive dose of less than 65 mg characterizes 16 and 18% of patients allergic to egg or peanut. Less than 30 mg of milk proteins characterizes 5% of those allergic to milk. For milk, egg, and peanut, 1% of patients have a very low threshold, about 1 mg. Such data emphasize the necessity of using detection tests with a sensitivity better than 10 parts per million. The modifications of allergenicity undergone by protein ingredients that are now commonly introduced into industrially made products are not yet sufficiently known. A better knowledge of the reactive doses of these proteins is needed.

Allergens↗
PubMed1997

Nutritional evaluation of some commercial baby foods consumed in Saudi Arabia.

The nutritive value of six baby foods based on milk (Nido, Wadi Fatima and Gain) and milk-cereal blends (Cerelac with wheat, Cerelac with rice and Milupa 2) commonly used in the Kingdom of Saudia Arabia was evaluated chemically including fatty acids analysis and biologically in growing rats. The milk based products vs milk-cereal blends provided (per 100 kcal) protein (3.8-5.0 g vs 3.7-3.8 g), fat (5.2-5.7 g vs 2.0-4.8 g), available carbohydrates (7.3-9.5 g vs 10.5-16.6 g), Ca (159-189 mg vs 101-145 mg), Mg (15-18 mg vs 14-20 mg), Na (32-39 mg vs 42-51 mg), K (160-180 mg vs 122-144 mg), Fe (1.4-1.8 mg vs 1.5-1.9 mg), Cu (0.04-0.09 mg vs 0.09-0.1 mg), Zn (0.8-1.2 mg vs 0.8-1.1 mg), and linoleic acid (208-1343 mg vs 518-639 mg). Metabolizable energy (ME) values in milk based products (487-495 kcal/100 g) were higher than milk-cereal blends (404-473 kcal/100 g). The true protein digestibility (TD) varied from (93-95%) in milk based foods to (94-95%) in milk-cereal blends. The net protein utilization (NPU) ranged between (0.74-0.78) in milk based products and (0.68-0.74) in milk-cereal blends. The net dietary protein calorie percent (NDP cal%) was higher in milk based foods (11.7-15.0%) than milk-cereal blends (10.2-11.1%). An imbalance of calories and nutrients in some baby foods was noticed. However, the protein quality was satisfactory and could meet the protein requirements of infants and toddlers as indicated by NDP cal% values.

Animals↗
PubMed1995

Variations in lectin binding properties of intestinal M cells.

Glycoconjugate expression in follicle-associated epithelia has been examined by application of a panel of lectins to fixed, whole tissue preparations of mouse and rabbit Peyer's and caecal patches. Of those examined, only alpha-L-fucose-specific lectins such as Ulex europaeus agglutinin I bound selectively to M cells in murine Peyer's patches and this selectivity did not extend to mouse caecal patch M cells. In common with a variety of other lectins (wheat germ agglutinin, peanut agglutinin and Bandeiraea simplicifolia agglutinin II), Ulex europaeus agglutinin I exhibited selective binding to M cells in rabbit caecal patch and to enterocytes in rabbit Peyer's patch. The marked regional and species variation in M cell surface characteristics documented here has implications for the normal antigen-sampling function of M cells and to the rational design of particulates for transepithelial delivery via the M cell portal.

Animals↗