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Microbiology of wheat and flour milling in Australia.

A survey was undertaken to determine the microbiological status of Australian wheat and the distribution of microorganisms in the flour milling fractions and end products. A total of 650 milling process and end product samples was obtained from nine flour mills located in New South Wales (4), Queensland (2), Victoria (2) and Western Australia (1) during the 1997-1998 and 1998-1999 wheat seasons. Most frequent (modal) counts in wheat and flour were, respectively, as follows: aerobic mesophilic plate count, 10(5) and 10(2) colony forming units/gram (cfu/g); coliforms, 10 and 1 most probable number/gram (MPN/g); Bacillus spp., 10(4) and 10(2) cfu/g; B. cereus, 1 and 0.1 MPN/g; mesophilic aerobic spores, 10 and 1 cfu/g; aerobic thermophiles, both 10 cfu/g; yeasts, 10(3) and 10(2) cfu/g, and moulds, 10(3) and 10(2) cfu/g. Bacillus spp., coliforms, yeasts and moulds were the most frequently detected microorganisms throughout the survey. The most common moulds isolated were Aspergillus, Penicillium, Cladosporium and Eurotium spp. Environmental serovars of Salmonella were isolated from two samples. Escherichia coli and B. cereus were present at very low levels, a majority of positive samples being at the minimum level of detection (3 and 0.3 MPN/g, respectively). As wheat grain layers are separated, surface-adhering contaminants are concentrated in end product bran, wheat germ and pollard, which comprise the outer layers of the grain. Consequently, the inner endosperm fraction contains lower microbial counts, and flour is the cleanest end product of the milling process. Higher microbiological counts midstream in the milling process indicate that equipment contamination may contribute to microbiological contamination; however, the microbiological quality of incoming wheat has a strong influence on the ultimate quality of milling end products.

Australia↗

Export of mRNA from microinjected nuclei of Xenopus laevis oocytes.

Export of mRNA from the nucleus to the cytoplasm was studied in mature Xenopus laevis oocytes. In vitro transcribed, capped 32P-labeled mRNA was microinjected into nuclei, and its appearance in the cytoplasm measured by counting radioactivity or by RNA extraction and gel electrophoresis. Both for a 5.0-kb transferrin receptor mRNA and a 2.0-kb 4F2 antigen heavy chain mRNA we found saturable transport with an apparent Km of 3.6 x 10(8) molecules per oocyte nucleus. Under non-saturating conditions the half-time for mRNA export from the nucleus was approximately 2 min at 20 degrees C. At higher concentrations of injected mRNA this half-time was prolonged, and the maximal transport rate was reached at approximately 1.6 x 10(8) molecules/min. mRNA transport showed properties of an energy-dependent mechanism, since it was inhibited at 4 degrees C or by ATP depletion. Co-injection of the cap dinucleotide m7GpppG blocked the export effectively, suggesting a role for the cap in this process. The export was also inhibited by the pre-injection of wheat germ agglutinin. The effect of the lectin was specific and abolished by co-injection of N-acetylglucosamine. Finally, we found significant competitive inhibition in mRNA export by the presence of tRNA. Our results suggest that mRNA transport is a facilitated process which may share common steps with tRNA transport. Preliminary gel retardation experiments show that injected mRNA associates with endogenous nuclear proteins and suggest an exchange of some of the bound components during the transport to the cytoplasm.

Adenosine Triphosphate↗

Dietary fiber values for various breads are higher using enzymatic analysis rather than detergent analysis.

OBJECTIVE: To compare the dietary fiber content of breads commonly eaten in Georgia, Michigan, Nevada, and Vermont. DESIGN: Dietary fiber content was determined according to neutral detergent fiber (NDF) plus pectin values and enzymatic gravimetric fiber values. White, whole-wheat, 100% stone-ground whole-wheat, cracked-wheat, and Roman Meal breads were analyzed for hemicellulose, cellulose, lignin, total NDF, pectin, and enzymatic gravimetric total fiber. STATISTICAL ANALYSES PERFORMED: Analysis of variance was used to determine significant F tests, and individual means were compared using Duncan's multiple range test. RESULTS: The 100% stone-ground whole-wheat bread had the most total fiber and fiber components followed by whole-wheat, Roman Meal, cracked-wheat, and white bread, except for pectin, of which Roman Meal had the most. The enzymatic gravimetric method of determining total fiber gave significantly (P < .0001) greater dietary fiber values than the values obtained by analyzing NDF plus pectin. These values ranged from 153% to 370% higher; however, there was a high degree of correlation (r = .95) between the methods. CONCLUSIONS: When recommending types of breads, dietitians should be aware of added ingredients that influence fiber values. When using published fiber values, dietitians should consider the technique used to quantify the fiber content.

Bread↗

Food allergies and migraine.

60 migraine patients completed elimination diets after a 5-day period of withdrawal from their normal diet. 52 (87%) of these patients had been using oral contraceptive steroids, tobacco, and/or ergotamine for an average of 3 years, 22 years, and 7.4 years respectively. The commonest foods causing reactions were wheat (78%), orange (65%), eggs (45%), tea and coffee (40% each), chocolate and milk (37%) each), beef (35%), and corn, cane sugar, and yeast (33% each). When an average of ten common foods were avoided there was a dramatic fall in the number of headaches per month, 85% of patients becoming headache-free. The 25% of patients with hypertension became normotensive. Chemicals in the home environment can make this testing difficult for outpatients. Both immunological and non-immunological mechanisms may play a part in the pathogenesis of migraine caused by food intolerance.

Adolescent↗

Relationship between IgG1 and IgG4 antibodies to foods and the development of IgE antibodies to inhalant allergens. I. Establishment of a scoring system for the overall food responsiveness and its application to 213 unselected children.

To obtain reference levels for subsequent investigations, we analysed the IgG1 and IgG4 antibody levels to common foods in the sera of 213 unselected children (age 3 months to 14 years). The children were clustered into five age groups and tested on a broad screening panel of common foods. We used the IgG1 and IgG4 RAST with Sepharose-coupled antigens: cows' milk, hens' egg white, banana, legumes (a mixture of soybean and peanut), grains (a mixture of wheat and rice), potato, orange and pork. In all age groups and all antigens, a considerable variability in the antibody response was found. As for some assays more than half of the sera were negative or borderline, statistics based on interval or ordinal scaling were considered inappropriate and we resorted to nominal classification. We decided to use, for each of the assays, the 75-percentile of the age group as a cut-off level. Each antibody titre was thus converted into positive (more than the 75-percentile of that age group) or negative; the number of positive tests was used as the score. This resulted in a sigma G1-score and a sigma G4 score (summed scores for IgG1 and IgG4 antibodies, respectively). The results of the present study indicate that children with a high response to one food tend to have elevated responses to other non-related foods, possibly explained by a defective mucosal barrier and/or a hyperactive immune system. This suggests that a high-food responder phenotype may exist.

Adolescent↗

Wheat embryo ribonucleates. V. Generation of N2--dimethylguanylate when 'fully sequenced' homogeneous species of transfer RNA are used as substrates for wheat embryo methyltransferases.

When S-adenosly[methyl-14-C]methionine and various species of transfer RNA are used as substrates for wheat embryo methyltransferases, the principal site of guanylate-N-2 methylation can be shown to be a G-residue between the stems of the dihydrouridine and anticodon loops. This common site of guanylate-N-2 methylation is referred to as the interstem target site. 2. When the interstem target site is the non-terminal G-residue in a G-C-G-C sequence, as in the cases of Escherichia coli tRNA1-Leu, tRNA-Ile, and tRNA3-Ser, there is preponderant dimethylation to yield N-2-dimethylguanylate. 3. When the interstem target site is part of a U-C-G-U sequence, as in the case of E. coli tRNAf-Met, there is diminished dimethylation and correspondingly increased monomethylation to yield N-2-monomethylguanylate. 4. When the interstem target site is the non-terminal G-residue in an A-U-G-G sequence, as in the case of yeast tRNA-Asp, there is negligible dimethylation and almost exclusive monomethylation to yield N-2-monomethylguanylate. 5. The concerted way in which the primary, secondary, and tertiary structures of tRNA molecules might influence the efficacy of these methylations is the subject of a brief discussion. Attention is also focused on the evolutionary and molecular basis for the generally non-random distributions of methylated oligonucleotide sequences in ribosomal and transfer ribonucleates.

Base Sequence↗

Celiac disease.

Celiac disease is an autoimmune disease that occurs in genetically predisposed individuals as the result of an immune response to gluten. This immune response occurs in both the lamina propria and the epithelium of the small intestine. There is a close link to HLA DQ2 and DQ8, although these HLA genes account for only 40% of the genetic influence. Environmental factors, such as the amount and timing of gluten administration in infancy, as well as breastfeeding, influence the disease. Serologic screening studies that use sensitive and specific antibody tests have revealed the disease to be common, occurring in approximately 1% of the population. Clinical presentations are diverse and atypical; the majority of patients lack diarrhea. Therapy is a gluten-free diet that requires avoidance of wheat, rye, and barley, although there is potential for other therapies based on our understanding of the pathophysiology of the disease.

Celiac Disease↗

Ruminal ciliated protozoa in cattle fed finishing diets with or without supplemental fat.

Ruminal samples were collected at slaughter from 364 unfasted steers fed different finishing diets to obtain information on numbers and species distribution of ciliated protozoa in feedlot cattle. Total numbers of protozoa averaged 1.59 X 10(5)/g of ruminal contents. A total of 47 steers (12.9%) were defaunated, but 4.1% of the steers possessed numbers of protozoa greater than 10(6)/g. Entodinium species did not always dominate the protozoan populations; 41 faunated steers (11.2%) were devoid of entodinia, and 79 additional steers (21.7%) possessed populations dominated (greater than 50%) by other genera. Isotricha was the most commonly occurring genus supplanting Entodinium, but Polyplastron and Epidinium were frequently present in high concentrations. Tallow and soybean soapstock supplementation reduced (P less than .05) numbers of protozoa in steers consuming wheat diets. However, yellow grease supplementation did not affect numbers of protozoa in steers fed either sorghum or corn diets. Average ruminal pH was 6.20 on the wheat diet, 6.05 on the corn diet, and 5.69 and 6.23 for the two sorghum diets, respectively. We found no correlation between ruminal pH and numbers of protozoa on any diet. The presence of relatively high protozoan concentrations and few defaunated animals in feedlot cattle necessitates reevaluation of the role that ciliated protozoa play in ruminal metabolism of animals fed processed, high-concentrate diets.

Animal Feed↗

Celiac disease. CME update for family physicians.

OBJECTIVE: To review current understanding of the epidemiology, pathophysiology, diagnosis, and management of celiac disease. QUALITY OF EVIDENCE: Few recent randomized controlled trials (level I evidence) have studied treatments for celiac disease. There are recent comparative studies (level II evidence) and there is well established consensus (level III evidence) on diagnosis and treatment of celiac disease. MAIN MESSAGE: Celiac disease is an immune-mediated small bowel enteropathy caused by exposure to wheat gluten protein. The disease can be insidious and often presents with only subtle extraintestinal manifestations in a variety of organ systems. Recent epidemiologic surveys suggest celiac disease is much more common in North America than previously thought. Advances in immunology and screening have made diagnosis more reliable than in the past. Removing gluten from the diet is effective in most cases. CONCLUSION: Celiac disease manifests subtly and is an easy diagnosis to miss. Good laboratory screening tests and effective treatment are available. Family practitioners should consider celiac disease in patients who present with confounding symptoms.

Algorithms↗

[Pseudomonas genus bacteria on weeds].

It has been shown in the work that the weeds (couch-grass and ryegrass) may be affected by bacterial diseases in natural conditions, Pseudomonas genus bacteria being their agents. The isolated bacteria are highly-aggressive in respect of the host-plant and a wide range of cultivated plants: wheat, rye, oats, barley, apple-tree and pear-tree. In contrast to highly aggressive bacteria isolated from the affected weeds, bacteria-epi phytes isolated from formally healthy plants (common amaranth, orache, flat-leaved spurge, field sow thistle, matricary, common coltsfoot, narrow-leaved vetch) and identified as P. syringae pv. coronafaciens, were characterized by weak aggression. A wide range of ecological niches of bacteria evidently promote their revival and distribution everywhere in nature.

Agriculture↗

Fatal parathion poisoning in Sierra Leone.

In May and June 1986, 49 persons in Sierra Leone were acutely poisoned by the organothiophosphate insecticide, parathion. Fourteen people died. Illness occurred in three episodes at two different locations that were 44 km apart. A study of 21 cases and 22 household controls was undertaken to explore which factors were associated with the development of the symptoms. Cases were more likely than controls to have eaten bread in the 4 hours before becoming ill (odds ratio, 12.7; 95% confidence interval, 2.4-83.8). Scrapings of residue from the floor of the truck that had brought the wheat flour from the mill to the general store where the baker purchased it were positive for parathion, suggesting that the flour had been contaminated during transport. Pesticide poisoning is a common problem in the developing world, and public health measures such as restricting the use of parathion may help to prevent fatal poisonings.

Adolescent↗

Calcium intakes in individuals on diets for the management of cows' milk allergy: a case control study.

Three hundred and twenty-three individuals with self-reported food allergy were recruited by media advertisements. Questionnaire information was collected on all respondents. Chocolate (57%), milk (47%), wheat (36%) and food additives (35%) were the most frequently implicated foods. The most frequently reported symptoms were itching (43%), skin rash (43%) and tiredness (43%). Food avoidance was the most common form of food allergy diagnosis (33%) with only 8% of respondents reporting food challenge in food allergy diagnosis. Self-diagnosis was reported by 34% of respondents with 29% and 24% reporting diagnosis by a general practitioner or a homeopath, respectively. Twenty-four per cent of respondents 'always' avoided and a further 57% 'nearly always' avoided the implicated food(s). A group of 38 adults with self-reported 'milk allergy' was selected for further study. Dietary assessments, using the dietary history method, were carried out on this subgroup and on age-, sex- and occupation-matched controls. The results of the dietary assessments revealed that the 'milk allergy' group had significantly higher intakes of fibre, beta-carotene, vitamin C, vitamin E, iron and folic acid (P < 0.05) and significantly lower intakes of calcium (P < 0.002) in comparison to controls. Eighteen per cent of the group used milk alternatives. Thirty-four per cent of the 'milk allergy' group took calcium-containing supplements. Even after calcium supplementation, the mean calcium intake of those who completely avoided milk was unacceptably low (441 mg/d).

Adult↗

Life-threatening, recurrent anaphylaxis caused by allergy to gliadin and exercise.

BACKGROUND: Exercise-induced urticaria or anaphylaxis is regarded as a distinct form of physical allergy. In some patients the symptoms occur only after ingestion of various food products in connection with exercise. We have come across patients with cereal dependent exercise-induced anaphylaxis. OBJECTIVES: The purpose of the present study was to analyse the allergens in cereals responsible for the severe anaphylactic symptoms and to verify the test methods suitable for screening the patients with cereal dependent exercise-induced anaphylaxis. METHODS: The patients underwent skin-prick tests (SPT) with common inhalant and food allergens as well as with various cereal extracts. IgE-immunoblotting was used to identify the allergenic fractions. RESULTS: Five patients found positive in SPT with NaCl wheat suspension had IgE antibodies to wheat, rye, barley and oats, especially directed against the ethanolsoluble protein fractions in immunoblotting. No IgE antibodies were detected against other cereals. The patients had been unaware of any cereal allergy since anaphylaxis occurred only in association with exercise postprandially. The patients were directed to follow a gluten-free diet and have been free from symptoms, being able to continue their outdoor physical activities. CONCLUSION: Wheat gliadin and the corresponding ethanol-soluble proteins of taxonomically closely related cereals were found to be the allergens in cereal-dependent exercise-induced anaphylaxis. Skin-prick testing with NaCl wheat suspension was a simple and practical test to screen patients with this kind of occult, possibly life-threatening, allergy to cereals.

Adult↗

Role of gangliosides in the uptake and retrograde axonal transport of cholera and tetanus toxin as compared to nerve growth factor and wheat germ agglutinin.

Previous investigations have shown that tetanus toxin is transported retrogradely in all peripheral neurons whereas the transport of NGF is confined to adrenergic and sensory neurons. Other macromolecules with molecular weights and general physiochemical properties similar to NGF and tetanus toxin (e.g., cytochrome C, insulin, horseradish peroxidase and bovine serum albumin) are not transported to a detectable extent if injected in comparable molar concentrations. For tetanus toxin, which is transported in all peripheral neurons, it has be assumed that it's retrograde transport depends on properties common to all neurons. In view of the relatively high ganglioside content of the neurons and the high affinity of tetanus toxin for the trisialoganglioside GT1, we studied the influence of gangliosides on the retrograde transport of tetanus toxin as compared to NGF. We included into the study cholera toxin which is known to have a high affinity for the monosialoganglioside GM1 and wheat germ agglutinatinin, a lectin with specific affinity for glycoproteins with N-acetyl-glucosamine residues. Both cholera toxin and wheat germ agglutinin were transported efficiently in all peripheral neurons. Preincubation of 125I-cholera toxin with monosialoganglioside GM1 completely blocked its retrograde axonal transport. The transport of NGF and wheat germ agglutinin was affected neither by various purified gangliosides nor by a mixture of bovine brain gangliosides. The transport of tetanus toxin was only reduced by 50% both by the trisialoganglioside GT1 and the bovine ganglioside mixture.

Adrenergic Fibers↗

Identification of structural and secretory lectin-binding glycoproteins of normal and cancerous human prostate.

We have utilized the technique of lectin-loading of SDS gels with iodinated concanavalin A and wheat germ agglutinin to identify glycoproteins in prostatic and seminal fluids as well as in prostate tissue fractions. The following subunits which bound both lectins were detected: (a) 50, 43 and 38 kDa subunits common to prostatic and seminal fluids, and an additional 55 kDa subunit which predominates only in prostatic fluid; (b) 78, 55, 50 and 43 kDa subunits in prostatic tissue cytosol and (c) 195, 170, 135, 116 and 95 kDa subunits present in the particulate fractions of prostatic tissue. Immunoblotting using specific rabbit antibodies revealed the 50 kDa band to be prostatic acid phosphatase and the 38 kDa band to be prostate-specific antigen. Interestingly, antibodies directed toward prostatic acid phosphatase were found to cross-react with the 43 kDa band. Fractionation on sucrose gradients showed that several of these particulate glycoproteins were associated with a vesicle fraction enriched in adenylate cyclase activity, implying that they are plasma membrane glycoproteins. Comparison of soluble and particulate fractions of normal and cancerous tissue homogenates was made by densitometric scanning of autoradiograms of lectin-loaded gels. Similar relative intensities of lectin-binding were obtained for corresponding proteins in normal and cancerous tissue fractions. Also, immunoblotting showed no differences in prostatic acid phosphatase or prostate-specific antigen between normal and cancerous soluble homogenate fractions. Our results suggest that major lectin-binding proteins are conserved in the transition from normal to cancerous tissue. These results may be useful in developing a multiple-marker profile of metastatic prostate cancer and for the design of imaging agents, such as monoclonal antibodies, to prominent soluble and particulate prostate glycoproteins.

Acid Phosphatase↗

Chromosome mapping and identification of amphiphilic proteins of hexaploid wheat kernels.

Amphiphilic proteomic analysis was carried out on the ITMI (International Triticae Mapping Population) population resulting from a cross between "Synthetic", i.e.: "W7984" and "Opata". Out of a total of 446 spots, 170 were specific to either of the two parents, and 276 were common to both. Preliminary analysis, which was performed on 80 progenies (Amiour et al. 2002a), was completed here using a total of 101 selfed lines. Seventy two Loci of amphiphilic spots placed at LOD = 5 were conclusively assigned to 15 chromosomes. Some spots mapped during the first analysis were eliminated because of the significant distortion segregation observed in the second analysis. Group-1 chromosomes had by far the greatest number of mapped spots (51). Using the Quantitative Trait Loci (QTLs) approach, analysis of the quantitative variation of each spot revealed that 96 spots out of the 170 specific ones showed at least one Protein Quantity Locus (PQL). These PQLs were distributed throughout the genome. With Matrix Laser Desorption Ionisation Time Of Flight (MALDI-TOF) spectrometry and Database interrogation, a total of 93 specific and 41 common spots were identified. This enabled us to show that the majority of these proteins are associated with membranes and/or play a role in plant defence against external invasions. Using multiple-regression analysis, other amphiphilic proteins, in addition to puroindolines, were shown to be involved in variation in kernel hardness in the ITMI population.

Chromosome Mapping↗

Characterization of a Drosophila protein that binds both epidermal growth factor and insulin-related growth factors.

The identification of a novel protein from Drosophila melanogaster that binds both mammalian epidermal growth factor (EGF) and insulin has been reported (Thompson, K. L., S. J. Decker, and M. R. Rosner, 1985, Proc. Natl. Acad. Sci. USA., 82:8443-8447). This 100-kD protein (designated dp100) is also recognized by an antiserum against the human EGF receptor. To further characterize the properties of this protein, we have determined the binding spectrum, glycosylation state, and cellular distribution of dp100. Our results indicate that dp100 binds to other insulin-like and EGF-like growth factors with dissociation constants ranging from 10(-6) to 10(-9) M, and these ligands compete with each other for binding to dp100. All other ligands tested, including platelet-derived growth factor, transforming growth factor-beta, nerve growth factor, and glucagon, either did not bind or bound with a Kd greater than 10(-6) M. Unlike the Drosophila insulin receptor, dp100 does not bind to wheat germ agglutinin and is present in a cytoplasmic as well as a membrane-bound form that cannot be differentiated by two-dimensional PAGE. Further, dp100 is the sole transforming growth factor-alpha-binding protein detected by affinity labeling in Drosophila Kc cells. These results indicate that dp100 shares properties in common with, but distinct from, the Drosophila homologues of the insulin and EGF receptors.

Animals↗

Genetic structure of an aphid studied using microsatellites: cyclic parthenogenesis, differentiated lineages and host specialization.

In a previous study, samples of the grain aphid Sitobion avenae (F.) were collected from wheat and adjacent cocksfoot hosts in a population thought to be primarily parthenogenetic, and DNA from individual aphids was analysed with a multilocus technique. Here we have applied single-locus microsatellites and a mitochondrial DNA marker to a subset of the same DNA extracts, and have made several additional inferences about important genetic and population processes in S. avenae. Microsatellite analysis indicated very high levels of genic and genotypic variation. S. avenae fell into three genotypic groups inferred to be almost noninterbreeding, while analysis of linkage and Hardy-Weinberg equilibria suggested high levels of sexual recombination within each genotypic group. Host specialization was evident: one lineage was found only on wheat, and one (bearing many alleles inferred to be introgressed from the blackberry-grass aphid S. fragariae (Walker)) was found only on cocksfoot. The third group of interrelated genotypes was found commonly on both hosts. Although most genotypes were found only once, some were much more numerous in the sample than expected from the frequency of the alleles they contained. This, and rapid temporal changes in genotypic composition of samples, indicates strong selective differences between genotypes and lineages. In the major genotypic group, the commonest genotypes were significantly more homozygous than were rare ones: thus these data may help to explain the frequent observation of homozygous excess in aphid allozymes. The genotype group showing S. avenae-like as well as S. fragariae-like alleles also carried S. fragariae-like mitochondrial DNA in at least 25/31 cases, indicating gender-asymmetrical hybridization.

Animals↗