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A Nuclear Magnetic Resonance Study of Water in Cold-acclimating Cereals.

Continuous wave nuclear magnetic resonance (NMR) studies indicated that the line width of the water absorption peak (Deltav(1/2)) from crowns of winter and spring wheat (Triticum aestivum L.) increased during cold acclimation. There was a negative correlation between Deltav(1/2) and crown water content, and both of these parameters were correlated with the lowest survival temperature at which 50% or more of the crowns were not killed by freezing (LT(50)). Regression analyses indicated that Deltav(1/2) and water content account for similar variability in LT(50). Slow dehydration of unacclimated winter wheat crowns by artificial means resulted in similarly correlated changes in water content and Deltav(1/2). Rapid dehydration of unacclimated crowns reduced water content but did not influence Deltav(1/2). The incubation of unacclimated winter wheat crowns in a sucrose medium reduced water content and increased Deltav(1/2). The increase in Deltav(1/2) appears to be dependent in part on a reduction in water content and an increase in solutes.Longitudinal (T(1)) and transverse (T(2)) relaxation times of water protons in cereals at different stages of cold acclimation were measured using pulse NMR methods. The T(1) and T(2) signals each demonstrated the existence of two populations of water, one with a short and one with a long relaxation time. During the first 3 weeks of acclimation, the long T(2) decreased significantly in winter-hardy cereals, and did not change in a spring wheat until the 5th week of hardening. There was no change in the long T(1) until the 3rd week of hardening for the winter cereals and until the 7th week of hardening for the spring wheat. No simple relationship could be established between T(1) or T(2) and cold hardiness. Neither continuous wave or pulsed NMR spectroscopy can be used as a diagnostic tool in predicting the cold hardiness of winter wheats. An increase in Deltav(1/2) or a reduction in relaxation times does not provide evidence for ordering of the bulk of the cell water.

Journal Article↗

Pyruvate orthophosphate dikinase from the immature grains of cereal grasses.

Pyruvate orthophosphate dikinase has been identified in the green grains of eight cereal grasses, most of which are classified as C(3) plants. The wheat (Triticum aestivum L. cv. Lerma Rojo) grain enzyme was further investigated: activity was low in very young grains, increased to a maximum at about 25 days after anthesis, then returned to a low level as the grain matured. It appeared to be located in the aleurone layer. A procedure was developed for obtaining partially purified preparations of pyruvate orthophosphate dikinase from the ears of wheat, oat (Avena sativa L.), barley (Hordeum distichum L.), and rye (Secale cereale L.). These preparations were suitable for measuring activities in both the forward and reverse reaction directions. The affinities of these enzymes for the six substrates (pyruvate, orthophosphate, and ATP in the forward reaction; phosphoenolpyruvate, pyrophosphate, and AMP in the reverse reaction) were determined and found to be similar to the reported affinities of the enzyme from the leaves of the C(4) plant Zea mays. A possible role for pyruvate orthophosphate dikinase in cereal grains is considered briefly.

Journal Article↗

Comparisons of Peptide hydrolase activities in cereals.

Carboxypeptidase activity (hydrolysis of N-carbobenzoxy-l-phenylalanyl-l-alanine) is high in a number of temperate zone cereals, originating in Asia Minor (wheat, barley, oats, wild oats, rye, triticale) compared to other cereals originating in central America or Asia (maize, sorghum, rice). However, endopeptidase activity (hydrolysis of azocasein or hemoglobin) is relatively much higher in the latter group. Comparison of trichloroacetic acid (TCA)-soluble products derived from the hydrolysis of hemoglobin showed that carboxyterminal amino acids (histidine, arginine, and tyrosine), are released when extracts from wheat and barley endosperms are used. With extracts from corn endosperms, much more TCA-soluble ultraviolet- absorbing material is released, but very little is released as free amino acids within the first 2 hours and the expected C-terminal amino acids of hemoglobin are not detected in significant amounts. These results suggest that the method of hydrolysis of the storage proteins may be significantly different in these two classes of cereals.

Journal Article↗

Physical properties and sixth graders' acceptance of an extruded ready-to-eat sweetpotato breakfast cereal.

Extruded ready-to-eat breakfast cereals (RTEBCs) were made from varying levels of sweetpotato flour (SPF), whole-wheat bran (WWB), and extrusion cooking. Moisture, protein, and ash contents were lower in the 100% SPF than the 100% WWB. Carbohydrate, beta-carotene, and ascorbic acid contents were higher in the 100% SPF. Fat, thiamin, riboflavin contents, bulk densities, and the water absorption index were similar for the cereals. However, the expansion ratio was highest in the 100% SPF cereal. The 100% WWB had the lightest color and most fibrous morphology. Extruded RTEBC containing 100% SPF and 75%/25% SPF/WWB were well-liked and acceptable to sixth graders attending an elementary school in Auburn, Alabama, but the 100% WWB was unacceptable.

Child↗

Predicting noncompliant levels of ochratoxin A in cereal grain from Penicillium verrucosum counts.

AIMS: To model the probability of exceeding the European legislative limit of 5 microg ochratoxin A (OTA) per kilogram grain in relation to Penicillium verrucosum levels and storage conditions, and to evaluate the possibilities of using P. verrucosum colony counts for predicting noncompliant OTA levels. METHODS AND RESULTS: Cereal samples were inoculated with P. verrucosum spores and stored for up to 9 months at temperatures and water activities ranging from 10-25 degrees C and aw 0.77-0.95. A logistic regression analysis showed that the probability of exceeding 5 microg OTA kg(-1) grain was related to colony counts of P. verrucosum and water activity. The sensitivity and specificity of various P. verrucosum count thresholds for predicting noncompliant OTA levels were estimated, using data from the storage trial and natural cereal samples. CONCLUSION: The risk of exceeding 5 microg OTA kg(-1) grain increased with increasing levels of P. verrucosum, and with increasing water activities. A threshold of 1000 CFU P. verrucosum per gram grain is suggested to predict whether or not the legislative limit is exceeded. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has provided a tool to evaluate the levels of P. verrucosum in grain in relation to OTA levels. Hence, mycological analyses can be used to identify cereal samples with high risk of containing OTA levels above the legislative limit.

Avena↗

Neurolathyrism risk depends on type of grass pea preparation and on mixing with cereals and antioxidants.

OBJECTIVE: To study an array of household individual and dietary risk factors for neurolathyrism. METHOD: Case-control study using recent cases in a district highly affected by the recent neurolathyrism epidemic in Ethiopia: 108 households with cases and 104 households with no cases; 170 neurolathyrism cases, 370 intra-family controls and 170 community controls frequency matched for age and sex. RESULTS: A history of acute illness was associated with a two- and threefold increased risk of paralysis in community and intra-family controls, respectively. Soaking grass pea in water before cooking roughly halved the risk of neurolathyrism but cooking in clay utensils more than quadrupled it. Consumption of grass pea in the green unripe and boiled forms increased the risk 10 times or more. Mixing the food with gravy that contains condiments with antioxidant activity reduced it by a factor of 4. The consumption of grass pea mixed with cereals rich in sulphur amino acids was also highly protective, but the magnitude of the effect depended on the grass pea preparation consumed. CONCLUSION: Consumption of pure grass pea, especially in the green unripe and boiled forms, should be avoided. Communities at risk of neurolathyrism during famine crises should be encouraged to combine and use grass pea with cereals before the household cereal stock is fully depleted. Breeding programmes, alongside traditional attempts to reduce the toxin content, should enhance the content of sulphur amino acids and antioxidants in grass pea.

Adolescent↗

Monoclonal antibody-based enzyme-linked immunosorbent assay of Fusarium T-2 and zearalenone toxins in cereals.

Direct, competitive enzyme-linked immunosorbent assays (ELISAs) with monoclonal antibodies have been developed for quantitative determination of trichothecene T-2 toxin (T-2), and zearalenone (F-2) from different cereals: Among the several extraction solvents tried, 89% acetonitrile with additives was chosen. The extracts were then used without cleanup in the ELISA. With appropriate dilution of the samples (1:25 or 1:50), the matrix effects caused by lipid and/or protein content of the samples can be diminished to the extent that the assay is no longer impaired. The mean recoveries from cereals infected with 100 to 2,000 ng of T-2 and 50 to 500 ng of F-2 per g were 85 and 91%, respectively. The measuring range of the T-2 test is 100 to 2,000 ng/g, and that of the F-2 test is 25 to 400 ng/g. The mean within-assay and interassay coefficients of variation of standard curves are both less than 10%. According to recovery results with artificially infected cereals, our tests proved to be suitable for rapid screening of food and feed samples for the presence of T-2 and F-2 toxins.

Antibodies, Monoclonal↗

Analysis and sorting of rye (Secale cereale L.) chromosomes using flow cytometry.

Procedures for chromosome analysis and sorting using flow cytometry (flow cytogenetics) were developed for rye (Secale cereale L.). Suspensions of intact chromosomes were prepared by mechanical homogenization of synchronized root tips after mild fixation with formaldehyde. Histograms of relative fluorescence intensity obtained after the analysis of DAPI-stained chromosomes (flow karyotypes) were characterized and the chromosome content of the DNA peaks was determined. Chromosome 1R could be discriminated on a flow karyotype of S. cereale 'Imperial'. The remaining rye chromosomes (2R-7R) could be discriminated and sorted from individual wheat-rye addition lines. The analysis of lines with reconstructed karyotypes demonstrated a possibility of sorting translocation chromosomes. Supernumerary B chromosomes could be sorted from an experimental rye population and from S. cereale 'Adams'. Flow-sorted chromosomes were identified by fluorescence in situ hybridization (FISH) with probes for various DNA repeats. Large numbers of chromosomes of a single type sorted onto microscopic slides facilitated detection of rarely occurring chromosome variants by FISH with specific probes. PCR with chromosome-specific primers confirmed the identity of sorted fractions and indicated suitability of sorted chromosomes for physical mapping. The possibility to sort large numbers of chromosomes opens a way for the construction of large-insert chromosome-specific DNA libraries in rye.

Cell Separation↗

Gibberellins and flowering of grasses and cereals: prizing open the lid of the "florigen" black box.

Comprehensive studies in grasses show that gibberellins (GAs) play a role as a florigen. For Lolium temulentum, which flowers in response to a single long day (LD), GAs are a transmitted signal, their content increasing in the leaf early in the LD and then, hours later, at the shoot apex. There is a continuous trail of evidence of hormonal action of these GAs for L. temulentum and support for a similar role in the flowering of other LD-responsive temperate grasses and cereals. A characteristic of the initial flowering responses of grasses and cereals is their limited stem elongation. Interestingly, it is GAs with low effectiveness for stem elongation, GA5 and GA6, that reach the shoot apex and, structurally, are probably not degraded by 2-oxidase enzymes. By contrast, GA1 and GA4 cause stem elongation, may be inactive for floral evocation, and do not reach the vegetative shoot apex apparently because of susceptibility to degradation. However, GA4 can be florally active if protected against 2-oxidases either structurally or by using a 2-oxidase inhibitor. Later in inflorescence development, GA1 and GA4 can be detected at the shoot apex and are florally active if applied. The 2-oxidase restricting accessibility to the apex has probably declined at this time so there is a second florigenic, LD-regulated GA action. A growing body of molecular evidence supporting these actions of GA may provide a future basis for manipulating flowering of grasses and cereals.

Biological Evolution↗

Biology and management of economically important lepidopteran cereal stem borers in Africa.

Cereals (maize, sorghum, millet, rice) are extremely important crops grown in Africa for human consumption. Of the various insect pests attacking cereal crops in Africa, lepidopteran stem borers are by far the most injurious. All 21 economically important stem borers of cultivated grasses in Africa are indigenous except Chilo partellus, which invaded the continent from India, and C. sacchariphagus, which has recently been found in sugarcane in Mozambique. C. partellus is competitively displacing indigenous stem borers in East and southern Africa. A parasitoid, Cotesia flavipes, was introduced from Pakistan for biological control of C. partellus and caused a 32-55% decrease in stem borer densities. This article is an attempt to summarize the status of knowledge about economically important cereal stem borers in Africa with emphasis on their distribution, pest status and yield losses, diapause, natural enemies, cultural control, host plant resistance, and biological control. Special attention is given to Busseola fusca and C. partellus, the most important pests of maize and grain sorghum.

Africa↗

Supplementation of a cereal-milk formula with haem iron palliates the adverse effects of iron deficiency on calcium and magnesium metabolism in rats.

Aside from the well known alteration of Fe status in Fe deficiency, this condition has also a negative effect on the bioavailability of Ca and Mg. We studied the influence of the supplementation of a commercial cereal-milk formula with bovine blood on Fe, Ca, P, and Mg metabolism in control and Fe-deficient rats to investigate whether high Fe levels in diet produce some interactions and the possibility of decreasing these latter by a haem-Fe supplementation. The bioavailability in control and Fe-deficient animals was determined as the apparent digestibility coefficient and hemoglobin regeneration efficiency, both of which are accurate estimations of total Fe utilization. Non-fortified cereal-milk formula decreased the apparent digestibility coefficient of Ca and Mg in Fe-deficient rats; the concentrations of these minerals in liver, femur, and sternum were lower than in control animals. However, when the Fe content of the cereal-milk formula was doubled by supplementation with bovine blood, the adverse effects on the digestive utilization of Ca, and especially of Mg, were palliated, the concentration of these two minerals in the organs investigated increased, and the overall Fe status improved in Fe-deficient rats.

Anemia, Iron-Deficiency↗

Naturally developing antibodies to wheat gliadin fractions and to other cereal antigens in rabbits, rats and guinea pigs on normal laboratory diets.

Rabbits, rats and guinea pigs on normal laboratory diets were examined for gliadin and other cereal antibodies comparable to those found in the sera of human coeliac patients. Many of the rabbits had very high levels of gliadin antibodies. Rats showed much lower titres more comparable to the findings in normal human sera. In both these species the reactivity was directed principally against the alpha-gliadins. The spectrum of reactivity against the prolamines of other cereals was again comparable to that of human coeliac sera. A difference, however, was the high reactivity with maize prolamine; this was decidedly lower in sera of coeliac patients. Guinea pigs were distinctive in having relatively no serum antibodies to wheat gliadin and also minimal reactivity to the albumins, globulins and glutenins of wheat. Normal guinea pigs did, however, have antibodies to oat and maize prolamines. Following parenteral injection with total ethanol-soluble wheat gliadin, guinea pigs developed high-titred antibodies, not only to wheat gliadin fractions 1-8 (alpha-, beta-, gamma- and omega-gliadins), but also to all the cereal prolamines except those of rice. These investigations are steps in the evaluation of the role of circulating gliadin antibodies in gluten-sensitive enteropathy.

Animals↗

Ready-to-eat cereals and dental caries in children: a three-year study.

In a three-year study of 145 adolescent males in a boarding school in Wisconsin, there were no statistically significant differences in dental caries experience between a group consuming phosphates added to pre-sweetened and non-presweetened ready-to-eat cereals, and the control group, or between cereal eaters and non-cereal eaters.

Adolescent↗

Lack of bioequivalence of levofloxacin when coadministered with a mineral-fortified breakfast of juice and cereal.

Previous studies have demonstrated that the chelation interactions demonstrated between fluoroquinolones and antacids also occur when they are coadministered with mineral-fortified foods. This study was conducted to evaluate the bioequivalence of levofloxacin when administered in a fasting state as compared to when it was administered with a common breakfast of calcium-fortified orange juice and ready-to-eat cereal. Fourteen of 16 healthy volunteers completed this study and received 500 mg of levofloxacin with each of the following: (1) 12 ounces of water, (2) subject-measured portions of juice and cereal, and (3) subject-measured portions of juice and cereal with milk. Plasma samples were collected prior to dosing and for up to 48 hours after. The results demonstrated that neither fed phase was bioequivalent to the fasting arm in terms of Cmax (with milk, 79.2% [72.6%, 85.7%]; without milk, 79.1% [73.3%, 84.9%]). In addition, a weak correlation was identified between the amount of change in 24-hour exposure and mineral fortification. The results of this study further demonstrate a need to require additional fed-fasted bioequivalence studies for drugs that demonstrate no interaction with the FDA meal but have significant interactions with drugs or supplements that contain large amounts of multivalent ions.

Adult↗

Influence of CSP 310 and CSP 310-like proteins from cereals on mitochondrial energetic activity and lipid peroxidation in vitro and in vivo.

BACKGROUND: The development of chilling and freezing injury symptoms in plants is known to frequently coincide with peroxidation of free fatty acids. Mitochondria are one of the major sources of reactive oxygen species during cold stress. Recently it has been suggested that uncoupling of oxidation and phosphorylation in mitochondria during oxidative stress can decrease ROS formation by mitochondrial respiratory chain generation. At the same time, it is known that plant uncoupling mitochondrial protein (PUMP) and other UCP-like proteins are not the only uncoupling system in plant mitochondria. All plants have cyanide-resistant oxidase (AOX) whose activation causes an uncoupling of respiration and oxidative phosphorylation. Recently it has been found that in cereals, cold stress protein CSP 310 exists, and that this causes uncoupling of oxidation and phosphorylation in mitochondria. RESULTS: We studied the effects of CSP 310-like native cytoplasmic proteins from a number of cereal species (winter rye, winter wheat, Elymus and maize) on the energetic activity of winter wheat mitochondria. This showed that only CSP 310 (cold shock protein with molecular weight 310 kD) caused a significant increase of non-phosphorylative respiration. CSP 310-like proteins of other cereals studied did not have any significant influence on mitochondrial energetic activity. It was found that among CSP 310-like proteins only CSP 310 had prooxidant activity. At the same time, Elymus CSP 310-like proteins have antioxidant activity. The study of an influence of infiltration by different plant uncoupling system activators (pyruvate, which activates AOX, and linoleic acid which is a substrate and activator for PUMP and CSP 310) showed that all of these decreased lipid peroxidation during cold stress. CONCLUSIONS: Different influence of CSP 310-like proteins on mitochondrial energetic activity and lipid peroxidation presumably depend on the various subunit combinations in their composition. All the plant cell systems that caused an uncoupling of oxidation and phosphorylation in plant mitochondria can participate in plant defence from oxidative damage during cold stress.

Carrier Proteins↗

An in vitro animal model for the study of cereal components toxic in celiac disease.

Peptic-tryptic-Cotazym (PTC) digests were obtained, simulating in vivo protein digestion, from rice, maize, rye, oats, barley, and sorghum prolamines and tested on small intestine cultures from rat fetus. The PTC digests of the prolamine fractions from rice and maize, even when tested at a concentration as high as 0.5 mg/ml, did not affect in vitro differentiation and maturation of fetal rat jejunum that took place in vitro in a way comparable to what happens in vivo. On the contrary, the PTC digests of prolamines from rye, oats, barley, and sorghum were very active in slowing down in vitro development of fetal rat intestine. These results further strengthen earlier findings and all together show that there is a strong correlation between toxicity results of cereal and/or cereal components assessed with clinical trials or in vitro systems based on bioptic specimens of intestinal mucosa from celiac patients and with the culture of rat fetal intestine. Therefore, the rat fetal intestine culture is considered to be an adequate model for screening and investigating cereal peptides which are toxic for the celiac small intestinal mucosa.

Animals↗

Protein extraction from cereal seeds.

Seeds may contain different components such as starch and complex carbohydrates that can seriously reduce protein extraction. The proteins in cereal seeds are usually classified in four groups according to their solubility criteria: albumins, globulins, prolamins, and glutelins. They can be specifically extracted. A general procedure for extracting the proteins present in green seeds or immature cereal kernels is given. Then several procedures mostly adapted to cereal seeds are reported for: (1) the whole storage proteins (mostly prolamins and glutelins); (2) the albumins-globulins extracted using salt buffer; (3) the amphiphilic proteins extracted using a phase partitioning process; and (4) the proteins strongly attached to or within the starch granules of the seed endosperm. These procedures have been used for 2-D electrophoresis and proteomic analyses.

Chemical Precipitation↗

Ochratoxin a contamination of cereal grains and coffee in Hungary in the year 2001.

Ochratoxin A (OTA) is a nephrotoxic and carcinogenic mycotoxin, a secondary metabolite produced by mould fungi belonging to several Aspergillus and Penicillium species. It is formed during the storage of cereal grains and other plant-derived products. OTA ingested by humans and animals with the food or feed may exert deleterious effects on health. The purpose of this study was to investigate the ochratoxin contamination of the most important potential sources of OTA. The OTA content of cereal samples for human consumption (36 baking wheat, 16 wheat flour and 6 maize coarse meal samples) and feed grain samples (30 feeding wheat, 32 feeding maize and 20 feeding barley samples) collected in the mid-phase or at the end of the storage period and of 50 commercial coffee samples was determined. The analyses were performed by immunoaffinity column--high-performance liquid chromatography (IAC-HPLC). The limit of detection of the method was 0.1 ng/g. Of the wheat samples intended for human consumption, 8.3% contained OTA at 0.29 ng/g on the average (OTA ranges: 0.12-0.5 ng/g; Table 2). The OTA contamination of wheat flour and maize meal samples for human consumption was similar to that of the baking wheat samples. OTA contamination was found in 26.7% of the feeding wheat, 15.6% of the feeding maize and 35% of the feeding barley samples. The average values and the ranges of OTA levels found in the above samples were 12.2 and 0.3-62.8 ng/g, 4.9 and 1.9-8.3 ng/g, and 72 and 0.14-212 ng/g, respectively (Table 3). Sixty-six percent of the coffee samples were contaminated with OA (average level: 0.57 ng/g, ranges: 0.17-1.3 ng/g; Table 4). OTA contamination of baking wheat samples was found to be relatively low, presumably as a result of the favourable weather at harvest and the optimal storage conditions. Calculations made on the basis of the obtained results show that the daily OTA intake of an adult human from edible cereals is only 6.7 ng, while the amount taken up with coffee is 4.1 ng daily. The high prevalence and high levels of OTA contamination in feed grains can be explained by the unfavourable storage conditions, and this finding suggests that OA-related health problems may arise in animals, and that foods of animal origin may be contaminated with this mycotoxin.

Animal Feed↗