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Sequential extraction and quantitative recovery of gliadins, glutenins, and other proteins from small samples of wheat flour.

Methods to sequentially extract and fractionate wheat flour proteins were evaluated to reliably quantify gliadins, glutenins, and albumins/globulins in single flour samples. Compositions of the resulting protein fractions were analyzed by RP-HPLC combined with SDS-PAGE. Unknown proteins were identified by mass spectrometry or N-terminal sequencing. The best separation and recovery of discrete albumin/globulin, gliadin, and glutenin fractions from the same flour sample was achieved by extraction with 0.3 M NaI in 7.5% 1-propanol followed by 2% SDS, 25 mM DTT in 25 mM TRIS, pH 8.0, and precipitation of the solubilized proteins with ammonium acetate/methanol followed by acetone. Average flour composition for the variety Butte86 was 10% albumin/globulin, 40% gliadin, and 48% glutenin. This method should be useful for determining flour composition in diverse samples and evaluating relationships between proteins and end-use functionality.

Chemical Precipitation↗

Proximate composition and functional properties of fullfat and defatted beniseed (Sesamum indicum L.) flour.

The proximate composition and functional properties of fullfat and defatted beniseed (Sesamum indicum L.) flour were evaluated. Functional properties studied were foam capacity and stability, water and oil absorption, bulk density, emulsion capacity and nitrogen solubility. Defatting increased the crude protein, ash, crude fiber, carbohydrate and mineral contents. Defatted flour showed comparatively better foam capacity and stability, water absorption and emulsion capacities but diminished bulk density and oil absorption capacity. Nitrogen solubility was pH dependent with a minimum at pH 4 and maximum at pH 8. Maximum nitrogen solubility (95%) was recorded for defatted flour while that for the fullfat flour was 60%. The proximate composition and functional properties of the samples suggest that beniseed flour would have useful application in fabricated foods.

Absorption↗

Studies on the baking properties of wheat: pigeonpea flour blends.

Pigeonpea flour was substituted at levels of 0, 5, 10, 15, 20, 25% to wheat flour and whole wheat meal for bread and Chapatti making, respectively. Blends were prepared up to 50% for cookie making. Increasing levels of pigeonpeas in the blends significantly increased the protein and mineral content of the baked products. The bread from 10% pigeonpea flour blend with 2-3% vital gluten and 0.5% SSL had high loaf volume and loaf quality. Blends containing 15% pigeonpea flour were acceptable for Chapatti and 30% pigeonpea flour with 0.25% SSL were acceptable for cookie making.

Bread↗

Chemical, sensory and rheological properties of porridges from processed sorghum (Sorghum bicolor), bambara groundnut (Vigna subterranea L. Verdc) and sweet potato (Ipomoea batatas) flours.

The chemical, sensory and rheological properties of porridges made from blends of sprouted sorghum, bambara groundnuts and fermented sweet potatoes were examined. Sorghum and bambara groundnuts were sprouted for 48 h while sweet potatoes were fermented for the same period. Blends were formulated from the processed ingredients in the ratio of 60:40:0, 57:42:1, 55:44:1 and 52:46:2 (protein basis) of sorghum, bambara groundnuts and sweet potatoes. Porridges were prepared from the composite flours and the traditional sorghum complementary food. Standard assay methods were used to evaluate the flours for nutrient composition. The porridges were also tested for sensory properties and viscosity. Processing increased the levels of most of the nutrients evaluated. Relative to the sorghum traditional complementary food, the composite flours had higher levels of lipids, protein, ash, crude fiber and minerals (p < 0.05). The porridges from the composite flours were generally liked slightly by the panelists and were about seven times less viscous than the porridge from the traditional sorghum complementary food. Use of the composite flours, particularly the 52:46:2 blend, as a traditional complementary food should be encouraged in Nigeria especially with the increasing cost of commercial complementary foods.

Arachis↗

Suppression of cell-mediated immunity following oral feeding of mice with palmyrah (Borassus flabellifer L) flour.

Inbred mice fed with a 40% mixture of palmyrah flour pellets showed a significantly reduced capacity to mount a delayed-type hypersensitivity (DTH) response to sheep red blood cells (SRBC). Maximal suppression of DTH was noted after 6 days of feeding but was detectable even after 2 days. Maximal suppression occurred when flour feed was maintained into the sensitization period (with SRBC). The suppressive effect was transferable to normal mice fed with pellets ad libitum by viable spleen cells from mice fed with the 40% flour pellet mixture for a period of 2 days. Further characterization of the cells which were able to transfer suppression showed them to be T cells, some of which possess the Ly-1 surface antigen. The cells appear to be negative for Ly-2 surface antigen. Histologically, no evidence of lesions was noted in the liver and other organs of recipient mice which received cells from flour-fed donors. We thus favour the interpretation that oral feeding of mice with palmyrah flour induced the generation of T suppressor cells which were able to suppress the DTH response to SRBC. The significance and implications of these findings are discussed.

Animals↗

Deoxynivalenol and other Fusarium toxins in wheat and rye flours on the Danish market.

Information on the contamination of Danish cereals and cereal products with Fusarium toxins is limited and the last survey is from 1984/1985. In the present study, the occurrence of deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and zearalenone (ZON) was investigated in flour of common wheat, durum wheat and rye. The samples were collected from 1998 to 2001 from both mills and the retail market in Denmark. A total of 190 flour samples were analysed for DON and NIV and about 60 samples for HT-2, T-2 toxin and ZON. DON was most frequently detected with an incidence rate of 78% over all samples for all years. The contamination level varied considerably from year to year, and for wheat and rye the highest incidence and DON concentrations were found in samples from the 1998 harvest. There were regular and heavy rainfalls in Denmark during the flowering period of the crops that year, and DON was found in all samples, with mean concentrations in wheat and rye flour of 191 microg kg(-1) (n=14) and 99 microg kg(-1) (n=16), respectively. Comparison of data from each harvest year showed higher contents of DON in samples of wheat (range 20-527 microg kg(-1)) than in rye (20-257 microg kg(-1)). Contents of NIV, HT-2 toxin and ZON in samples of wheat and rye were generally low, and even in positive samples the contents were close to the detection limit of the methods. The T-2 toxin was detected in only a few of the wheat samples and in low amounts. However, the toxin was found in about 50% of the rye samples collected during 1998-2000, with a mean content of 49 microg kg(-1) (n=25). Durum wheat flour showed the highest DON contamination level, and all samples (n=33) collected during 2000 and 2001 contained DON with means and medians above 1100 microg kg(-1). Over 70% of the samples contained more than 500 microg kg(-1) DON, and the highest observed concentration was 2591 microg kg(-1). The concentration of T-2 toxin in durum wheat flour was also high with five of the 10 analysed samples containing more than 100 g kg(-1).

Denmark↗

Final report on the safety assessment of Peanut (Arachis hypogaea) Oil, Hydrogenated Peanut Oil, Peanut Acid, Peanut Glycerides, and Peanut (Arachis hypogaea) Flour.

Peanut (Arachis Hypogaea) Oil is the refined fixed oil obtained from the seed kernels of Arachis hypogaea. Hydrogenated Peanut Oil, Peanut Acid, and Peanut Glycerides are all derived from Peanut Oil. Peanut Flour is a powder obtained by the grinding of peanuts. The oils and glycerides function in cosmetic formulations as skin-conditioning agents. The acid functions as a surfactant-cleansing agent, and the flour functions as an abrasive, bulking agent and/or viscosity-increasing agent. In 1998, only Peanut Oil and Hydrogenated Peanut Oil were reported in use. When applied to the skin, Peanut Oil can enhance the absorption of other compounds. Hepatic changes were noted at microscopic examination of rats fed diets containing 15% edible Peanut Oil for 28 days, although no control group was maintained and the findings were also noted in rats fed fresh corn oil. United States Pharmacopeia (USP)-grade Peanut Oil was considered relatively nonirritating when injected into guinea pigs and monkeys. Technical-grade Peanut Oil was moderately irritating to rabbits and guinea pigs and mildly irritating to rats following dermal exposure. This same oil produced reactions in < or = 10% of 50 human males. Peanut Oil was not an ocular irritant in rabbits. Peanut Oil, either "laboratory expressed" or extracted using a food-grade solvent, was not carcinogenic to mice. Peanut Oil exerted anticarcinogenic activity when tested against known carcinogens. Peanuts are the food most likely to produce allergic and anaphylactic reactions. The major allergen is a protein that does not partition into Peanut Oil, Hydrogenated Peanut Oil, Peanut Acid, and Peanut Glycerides. Aflatoxins can be produced in stored agricultural crops such as peanuts, but do not partition into the oils, acids, or glycerides. Manufacturers were cautioned to make certain that the oils, acids, and glycerides are free of aflatoxins and protein. Formulators were cautioned that the oils, acids, or glycerides may enhance penetration and can affect the use of other ingredients whose safety assessment was based on their lack of absorption. The available studies on Peanut Oil supported the conclusion that Peanut Oil, Hydrogenated Peanut Oil, Peanut Acid, and Peanut Glycerides are safe for use in cosmetic formulations. Peanut (Arachis Hypogaea) Flour, however, is sufficiently different from the above ingredients such that its safety can not be supported by studies using the oil. The additional data needed for Peanut (Arachis Hypogaea) Flour are (1) concentration of use; (2) chemical specifications (i.e., aflatoxin and protein levels); (3) method of preparation; and (4) contact urticaria and dermal sensitization at concentration of use. Although data on aflatoxin levels are sought, it is expected that concentrations of aflatoxin should comply with U.S. government stipulations. Absent the additional data, it was concluded that the available data are insufficient to support the safety of Peanut (Arachis Hypogaea) Flour for use in cosmetic products.

Administration, Cutaneous↗

Granary trial of protein-enriched pea flour for the control of three stored-product insects in barley.

A granary trial was conducted to evaluate the efficacy of protein-enriched pea flour against three common stored-grain insects, Sitophilus oryzae (L.), Tribolium castaneum (Herbst), and Cryptolestes ferrugineus (Stephens). Six 30-t farm granaries were filled with approximately 11 t of barley. The barley was either not treated, treated with protein-enriched pea flour at 0.1% throughout the entire grain mass, or treated at 0.5% throughout the top half of the grain mass. Adult insects were released in screened boxes (two insects per kilogram barley for S. oryzae and T. castaneum 1.4 insects per kilogram barley for C. ferrugineus). Barley was sampled four times during the 70-d trial. The number and mortality of adults and emerged adults in the samples were noted. Four kinds of traps, flight, surface-pitfall, probe-pitfall, and sticky-bar, were placed at different locations in the granaries to estimate the movement of insects. The 0.1% protein-enriched pea flour treatment reduced adult numbers of S. oryzae by 93%, T. castaneum by 66%, and C. ferrugineus by 58%, and reduced the emerged adults by 87, 77, and 77%, respectively. Treating the top half of the barley with 0.5% protein-enriched pea flour had similar effects as treating the entire grain mass with 0.1% pea-protein flour. However, the top-half treatment failed to prevent insects from penetrating into the untreated lower layer. Differences between traps are discussed.

Animals↗

Protein-enriched pea flour extract protects stored milled rice against the rice weevil, Sitophilus oryzae.

Studies were conducted to evaluate the effect of a protein-enriched pea (Pisum sativum var. Bonneville) flour extract against the rice weevil, Sitophilus oryzae in its repellency, toxicity, effect on fecundity, stability and sensory properties. Milled rice admixed with pea flour extract at 1% concentration significantly repelled S. oryzae. Mortality of S. oryzae was found to increase and fecundity was markedly suppressed, in rice treated with 1% pea flour extract. The toxicity and reproductive effects of the pea protein-enriched rice were found to be stable for a period of 5 months. The sensory characteristics of stored rice when eaten were not affected by the treatment with pea flour extract. This study indicates that the protein-enriched flour extract obtained from the Bonneville pea may be feasible to protect stored milled rice from insect attack.

Animals↗

Occupational histamine poisoning by fish flour: a case report.

Histamine poisoning due to inhalation and skin contact with fish products is rarely described in the literature. This study presents a case of occupational histamine poisoning by spoiled fish flour via inhalation, skin and eye contact. Shipments of fish flour transported in black or blue bags, depending on the source, were handled by 20 harbour workers. Ten workers handling blue bags developed allergy-like skin, eye, gastrointestinal, respiratory and cardiac symptoms within 30 min. Workers handling black bags were symptom-free, except for minimal eye irritation. After consultation with the Poison Control Centre histamine poisoning was suspected. The histamine content, as determined by thin-layer chromatography, was 10-fold higher in samples from the blue than from the black bags (510 mg/100 g flour compared with 50 mg/100 g flour, respectively). Part of the shipment was labelled as hazardous for human health with permission for further usage only under specific personal protective measures. It is suggested that the highest permissible levels of histamine in fish flour and similar products should be set and legally adopted.

Adult↗

Physicochemical and microbiological properties of selected rice flour-based batters for fried chicken drumsticks.

Rice flour-based batter (RFBB) formulations for chicken drumstick coating were developed as an alternative for traditional wheat flour-based batter (WFBB). Physicochemical properties and storage stability of selected RFBB were evaluated and compared to WFBB. Batter pickup of RFBB formulated in combination with oxidized corn starch and methylcellulose (MC) was not significantly different from that of WFBB. In contrast, batters with only rice and corn flour (60:40% flour weight) exhibited significantly higher pickup. Rice flour batter with 15% oxidized corn starch had the lowest batter pickup. All RFBB exhibited (P < 0.05) lower oil absorption than WFBB. The TBA values of RFBB and WFBB increased (P < 0.05) with increased frozen storage time at -40 C for 90 d. The RFBB with MC exhibited the lowest TBA values, whereas WFBB had the highest values. Microstructural analysis revealed that freezing caused structural deterioration of all batters, but the RFBB with MC exhibited less freezing tolerance than other samples. The total plate counts of immediately fried or frozen fried chicken stored for 90 d were less than 1 log cfu/g sample. The RFBB with 5% oxidized corn starch and MC can replace WFBB on fried drumsticks. Additionally, RFBB results in a healthier product due to lower fat absorption.

Absorption↗

Immune response to flour and dust mites in a United Kingdom bakery.

In a study of 279 United Kingdom bakery workers a high prevalence of immunological response to storage mites was found. To determine whether this was the consequence of exposure to storage mites in bakery work, a population of salt packing workers was examined as a comparison group not at occupational risk of exposure to storage mites. Forty two per cent of both groups were atopic (had a positive skin prick response greater than negative controls to D pteronyssinus, grass pollen, or cat fur by 2 mm or more) and 33% had an immediate skin prick test response to at least one of four storage mites (L destructor, G domesticus, T putrescentiae, A Siro). A higher percentage of the salt packing workers than the bakery workers had a positive radioallergosorbent test (RAST) (greater than or equal to 0.35 PRU) to D pteronyssinus and to the four storage mites. Logistic regression analysis identified atopy as the most significant variable for a positive skin test and RAST response to storage mites in both groups of workers. RAST inhibition was used to analyse extracted area and personal air samples. Analysis of static area samples for aeroallergen showed immunological identity with flour but L destructor was found in only one of seven exposed filters. The concentration of airborne flour was related to exposure rank of perceived dustiness and gravimetric measurement of total dust. Nineteen out of 32 filters from workers in jobs with higher dust exposure (rank >/=6) had a level of > 10 microgram/m(3) flour whereas this concentrations was exceeded in only one of 23 filters from workers in low dust exposure (< rank 6). It is concluded that storage mites are not of special significance in allergic responses in bakery workers. The development of immunological (and airway) responsiveness to inhaled flour dust is increased in those exposed to higher concentrations of airborne allergen, which appears to be predominantly flour and not storage mites.

Air Pollutants, Occupational↗

Work related symptoms, sensitisation, and estimated exposure in workers not previously exposed to flour.

Findings are presented from the initial cross sectional phase of a cohort study of employees exposed to flour in bakeries or mills. Of 401 eligible workers in seven sites 344 (86%) were surveyed; symptoms assessed by self completed questionnaire, and sensitisation measured by the response to skin prick tests, were related to intensity of exposure both to total dust and to flour aeroallergen. Among 264 subjects without previous occupational exposure to flour, work related symptoms which started after first employment at the site were related to exposure intensity, especially when exposure was expressed in terms of flour aeroallergen. The relations with eye/nose and skin symptoms were independent of atopic status and cigarette smoking. Positive skin test responses to mixed flour and to alpha amylase were also more frequent with increasing exposure intensity, although this was confounded by atopic status. There was only a weak association between symptoms and specific sensitisation.

Adult↗

Measurement of airborne flour exposure with a monoclonal antibody-based immunoassay.

BACKGROUND: The development of simple and standardised methods to measure airborne levels of workplace biological allergens is an important step in reducing the incidence of occupational asthma. Such a method would be useful for measuring wheat flour allergens which cause asthma in bakers. Measurement of allergen per se rather than total dust enables exposure to be better defined. METHODS: Monoclonal antibodies were produced, their specificity analysed by immunoblotting and then used to affinity-purify a putative flour allergen. The importance of this protein as an allergen was tested by RAST using sera from allergic bakers and it was identified by N-terminal sequencing. Suitable monoclonal antibodies were chosen to develop an enzyme-linked immunosorbent assay. Commercial baking flours and personal airborne dust samples were analysed using the immunoassay. RESULTS: A sensitive and specific monoclonal antibody-based enzyme-linked immunosorbent assay was developed to measure a wheat alpha-amylase inhibitor. The wheat alpha-amylase inhibitor content of bulk wheat flours was 0.124% (95% confidence limits 0.083-0.164%) and airborne levels in bakeries had a geometric mean of 744 ng/m3 (95% confidence limits 371-1,496 ng/m3). CONCLUSION: This assay is suitable for widespread use as the monoclonal antibodies and standards are well defined and potentially infinitely available. The assay therefore offers distinct advantages over those exposure assessment methods currently in use. Comparable results would be obtained by different investigators over a prolonged time period. The assessment of flour allergen exposure and the relationship with clinical response could then be investigated using a multi-centered approach.

Air Pollutants, Occupational↗

Lysine fortification of wheat flour improves selected indices of the nutritional status of predominantly cereal-eating families in Pakistan.

Wheat provides more than 50% of the protein and calorie intake of the population of Pakistan. Legumes and animal protein that could complement the amino acid pattern of wheat, in which lysine is the first limiting amino acid for utilization of protein, are not affordable by members of lower socioeconomic groups in developing countries. The purpose of the study was to determine whether lysine fortification of wheat flour would have a positive impact on populations consuming a predominantly wheat-based diet. A double-blind study was carried out for three months on the outskirts of Peshawar, Pakistan. Forty families received wheat flour fortified with lysine, and 40 families received wheat flour without lysine. Wheat provided 59% of the protein for men, 65% for women, and 58% for children. The weight and height of the children in both groups increased during the study, but the increase was significantly greater in the lysine group. Hemoglobin increased significantly in the women receiving lysine-fortified flour. Transferrin levels increased significantly in men, women, and children in the lysine group as compared with those in the control group. Prealbumin increased significantly in adults receiving additional lysine but decreased in children. Men, women, and children in the lysine-supplemented families had significant increases in CD4, CD8, and complement C3 as compared with controls. These results indicate that lysine fortification of wheat flour can significantly improve sensitive indicators of nutritional status in a population consuming a diet in which 58% to 65% of the protein, depending on age and sex, is supplied by wheat.

Adult↗

Lysine-fortified wheat flour improves the nutritional and immunological status of wheat-eating families in northern China.

The purpose of this study was to determine the impact of the fortification of wheat flour with lysine on selected health indicators among farm families obtaining 58% to 67% of their dietary protein from wheat. A man, a woman, and a child aged 5 to 12 years were studied from each of 88 families in a village near Huixian City, Henan Province, China. Half of the families received wheat flour fortified with 3 g of lysine per kilogram for three months, and the other half received wheat flour without fortification. The results showed a significantly greater gain in the height and weight of children receiving lysine-fortified wheat flour. Hemoglobin values were not affected. The mean prealbumin values of adult men and women were higher in those receiving lysine. The numbers of CD3 T cells increased significantly in women and children, as did the complement fraction C3 and IgG in men, IgA in women, and IgG, IgA, IgM, and C3 in children. These results indicate that lysine fortification of wheat flour can significantly improve some indicators of the nutritional status and immune function of family members consuming a wheat-based diet.

Adult↗

Consumption of hypoallergenic flour prevents gluten-induced airway inflammation in Brown Norway rats.

Brown Norway rats were immunized with gluten, and then fed a diet containing hypoallergenic flour or an amino acid mixture. The rats were then made to inhale a solubilized gluten to induce gluten-specific bronchial asthma. The antibody levels in the serum of rats were measured by ELISA, and cell counts were done on cytospin preparations of bronchoalveolar lavage fluid. Body weight was decreased after allergen challenge in rats fed the amino acid mixture but not in rats fed the hypoallergenic flour. Antibody levels in the serum were significantly lower in rats fed hypoallergenic flour than in those fed the amino acid mixture. Differential cell counts in the bronchoalveolar lavage fluid showed that the numbers of eosinophils, lymphocytes, and neutrophils were significantly lower in rats fed the hypoallergenic flour than in those fed the amino acid mixture. These results suggest that hypoallergenic flour actively suppresses the allergic reactions, probably by inducing oral tolerance.

Animals↗

Effects of peroxidase and hydrogen peroxide on the dityrosine formation and the mixing characteristics of wheat-flour dough.

The effects of adding hydrogen peroxide and peroxidase to wheat-flour dough on dityrosine formation and mixing characteristics were investigated. Dityrosine in wheat-flour dough was identified by HPLC with a fluorescence detector and by LC/MS/MS. Formation of dityrosine increased with the addition of hydrogen peroxide, and hydrogen peroxide plus peroxidase, to wheat-flour dough, while the addition of peroxidase had no effect on the amount of dityrosine formed. The mixing curve obtained by a doughgraph changed with the addition of hydrogen peroxide, and hydrogen peroxide plus peroxidase; the peak time was significantly delayed and the dough development time was extended. We found that dityrosine cross-links in wheat-flour dough increased with the addition of peroxidase plus hydrogen peroxide. It is thought that these cross-links can lead to polymerization of the proteins in wheat-flour dough.

Bread↗