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Aflatoxin B1 in common Egyptian foods.

Samples of common Egyptian foods (17 nuts and seeds, 10 spices, 31 herbs and medicinal plants, 12 dried vegetables, and 28 cereal grains) were collected from markets in Cairo and Giza. A portion of each sample was extracted with chloroform, and the concentrated extract was cleaned by passing through a silica gel column. Aflatoxin B1 was determined by reversed-phase liquid chromatography with UV detection. The highest prevalence of aflatoxin B1 was in nuts and seeds (82%), followed by spices (40%), herbs and medicinal plants (29%), dried vegetables (25%), and cereal grains (21%). The highest mean concentration of aflatoxin B1 was in herb and medicinal plants (49 ppb), followed by cereals (36 ppb), spices (25 ppb), nuts and seeds (24 ppb), and dried vegetables (20 ppb). Among nuts and seeds, the prevalence of aflatoxin B1 was highest (100%) in watermelon seeds, inshell peanuts, and unshelled peanuts. The lowest prevalence and concentrations were in hommos (garbanzo beans). The highest concentrations of aflatoxin B1 were detected in foods that had no potential for field contamination but required drying during processing and storage, such as pomegranate peel, watermelon seeds, and molokhia.

Aflatoxin B1↗

Diet and lifestyle guidelines and desirable levels of risk factors for the prevention of diabetes and its vascular complications in Indians: a scientific statement of The International College of Nutrition. Indian Consensus Group for the Prevention of Diabetes.

BACKGROUND: There has been a rapid increase in the prevalence of diabetes and cardiovascular disease in India, in association with rapid changes in diet and lifestyle. In adults, the prevalence of diabetes, hypertension and coronary artery disease is two- to threefold greater in the urban population than in rural populations; it is associated with modest insulin resistance in urban groups. METHODS: In response to a proposal by the International College of Nutrition that specialist experts should develop consensus recommendations for the prevention of chronic diseases, Indian specialists in diabetes and vascular disease have collaborated to produce guidelines relevant to the population of India. RECOMMENDATIONS: Because Indian urban populations have a modest increase in overweight and low rates of obesity in association with the rapid emergence of diabetes and cardiovascular risk, a body mass index of 21 kg/m2 should be considered safe, with a range of 19-23 kg/m2 acceptable; > 23 kg/m2 should be considered overweight, and > 25 kg/m2 should be taken to indicate obesity. A waist:hip ratio > 0.88 in males and > 0.85 in females should be considered to indicate central obesity, because the prevalence of coronary disease, hypertension and associated disturbances of insulin resistance are more common above these limits. For the prevention of vascular disease, there is general international consensus that the desirable serum concentration of cholesterol should be < 170 mg/dl (> 4.42 mmol/l), which may also be optimal for Indians; values between 170 and 200 mg/dl (4.42-5.2 mmol/l) should be considered borderline. The critical values for low density lipoprotein cholesterol may be < 90 mg/dl (ideal), 90-110 mg/dl (borderline high) and > 110 mg/dl (high) (< 2.32, 2.32-2.84 and > 2.84 mmol/l, respectively). Fasting triglycerides should be < 150 mg/dl (< 1.69 mmol/l) and high-density lipoprotein cholesterol > 35 mg/dl (> 0.9 mmol/l). The limit for the total energy derived from fat intake should be < 21%/day (7% each for saturated, polyunsaturated and mono-unsaturated fatty acids). The carbohydrate intake should provide more than 65% of daily energy, mainly from complex carbohydrates. A daily dietary intake of 400 g fruits, vegetables and legumes, 400 g cereals, in conjunction with 25 g soya bean or mustard or canola oils (rich in n-3 fatty acids) in place of fats rich in saturated fat, may be protective against diabetes and vascular disease. Moderate physical activity with the aim of burning 300 Kcal/day (> 1255 KJ/day), and cessation of tobacco and alcohol consumption, may provide an effective programme for prevention of diabetes and its vascular complications in Indians.

Adult↗

Lupine-induced anaphylaxis.

BACKGROUND: Legumes are one of the most common foods causing allergic reactions in children and adults. Cross-reacting antibodies are frequently demonstrated in this family but the real clinical cross-reactivity is uncommon. OBJECTIVE: To report a case of lupine-induced anaphylaxis and to elucidate in vivo and in vitro cross-reactivity with some legumes. METHODS: Skin prick test (SPT) with some legumes were performed. Cap-IgE, ELISA-IgE, and immunoblotting were carried out. Open oral challenges with some legumes were performed. Cross-reactivity was studied by ELISA and immunoblotting inhibition. RESULTS: The results demonstrated type-I hypersensitivity reactions with lupine and some other legumes. Cap-IgE with peanut was positive but the SPT and ELISA-IgE were negative and the patient tolerated a peanut challenge. ELISA inhibition revealed a partial inhibition (62%) using lupine as the solid phase. Partial inhibition was demonstrated by immunoblotting inhibition. Open oral challenge with peanut and green bean were negative but positive with pea. CONCLUSION: We present a lupine sensitized patient with positive SPT and in vitro cross-reactivity with other legumes. Clinical cross-reactivity progressively developed over a 5-year period. Discrepancies were found between the clinical aspect and in vitro study of peanut allergy. Factors determining the wide variability in cross-reactivity among individuals are still obscure.

Adult↗

The Effect of Pseudomonas putida Colonization on Root Surface Peroxidase.

Increased activities of peroxidase and indole 3-acetic acid (IAA) oxidase were detected on root surfaces of bean (Phaseolus vulgaris) seedlings colonized with a soil saprophytic bacterium, Pseudomonas putida. IAA oxidase activity increased over 250-fold and peroxidase 8-fold. Enhancement was greater for 6-day-old than for 4- or 8-day-old inoculated plants No IAA oxidase or peroxidase activities were associated with the bacterial cells. Native polyacrylamide gel electrophoresis demonstrated that washes of P. putida-inoculated roots contained two zones of peroxidase activity. Only the more anodic bands were detected in washes from noninoculated roots. Ion exchange and molecular sizing gel chromatography of washes from P. putida-colonized roots separated two fractions of peroxidase activity. One fraction corresponded to the anodic bands detected in washes of P. putida inoculated and in noninoculated roots. A second fraction corresponded to the less anodic zone of peroxidase, which was characteristic of P. putida-inoculated plants. This peroxidase had a higher IAA oxidase to peroxidase ratio than the more anodic, common enzyme. The changes in root surface peroxidases caused by colonization by a saprophytic bacterium are discussed with reference to plant-pathogen interactions.

Journal Article↗

Kahweol and cafestol: inhibitors of hamster buccal pouch carcinogenesis.

Kahweol and cafestol, two compounds extracted from green coffee beans, were tested for cancer chemopreventive activity. For the experiment, 60 hamsters were divided into three equal groups and placed on one of three diets. The animals in Group I received a normal diet, whereas the animals in Groups II and III received the same diet supplemented with a 50:50 mixture of kahweol and cafestol. The content of the kahweol and cafestol mixture in these two diets was 0.2 g/kg of food (Group II) and 2.0 g/kg of food (Group III). After the hamsters adjusted to their respective diets, 16 hamsters from each group were selected. The left buccal pouches of these animals were painted three times weekly with a 0.5% solution of 7,12-dimethylbenz[a]anthracene (DMBA) in mineral oil. The 12 remaining hamsters were used as controls. The left buccal pouches of these animals were painted three times weekly with mineral oil. After 13 weeks (39 applications) the hamsters were killed. Multiple tumors were common in animals treated with DMBA; however, the animals receiving kahweol and cafestol in the diet (2 g/kg of food) exhibited a 35% reduction in tumor burden. Further comparisons between Groups I and III showed that this reduction in tumor burden was due to a decrease in tumor number. The results for Group II were inconclusive. Some reduction in tumor number was found, but this was offset by an increase in the size of the tumors.

Animals↗

The influence of methods of production, purification and storage of monoclonal antibodies upon their observed specificities.

A panel of 38 monoclonal antibodies was exposed to a variety of physical conditions commonly used in their purification and storage. Reactivity with the homologous (immunising) antigen was reduced in a significant proportion (42%) of antibodies following these physical treatments. Sensitivity to particular physical treatments correlated with antibody subclass. In 3 cases the relative specificity of the antibody (as measured by cross-reactivity with 16 closely related antigens) was also altered. Following passage of the cloned hybridomas as ascites, 3 antibodies exhibited marked differences in specificity, which suggested the selection of different genotypes. The importance of documenting the methods of production, purification and storage of particular monoclonal antibodies used in comparative assay is stressed.

Animals↗

Oral acyclovir for the prevention of herpes-associated erythema multiforme.

Herpes simplex virus is the single most common precipitator of erythema multiforme. Typically, erythema multiforme lesions appear 10 to 14 days after a recurrent herpes simplex virus infection and attacks can be disabling when they occur at frequent intervals. Prior to the introduction of acyclovir (Zovirax), there was no effective therapy to prevent herpes-associated erythema multiforme. Four patients were treated with a maintenance dose of acyclovir for periods ranging from 10 to 26 months; there were no significant side effects from the drug and only one recurrence of erythema multiforme. Oral acyclovir may become the treatment of choice for herpes-associated erythema multiforme.

Acyclovir↗

Structure of the trypsin-binding domain of Bowman-Birk type protease inhibitor and its interaction with trypsin.

The crystal structure of the complex formed by bovine trypsin and Bowman-Birk type protease inhibitor AB-I extracted from azuki beans (Vigna angularis) 'Takara' has been analyzed. The structure was solved by the application of the phase combination of single isomorphous phases and trypsin model phases, followed by phase improvement using the iterative Fourier technique. From the resulting electron density map, a three-dimensional atomic model of the trypsin binding domain of AB-I has been built. The peptide chain at the trypsin reactive site turns back sharply at Pro29 and forms a 9-residue ring (Cys24-Cys32). The 'front side' of this ring, consisting of the reactive site (Cys24-Met28), interacts with trypsin in a similar manner to other families of inhibitors and forms a stable complex, which seems to be maintained by the interactions with the 'back side' of this ring (Pro29-Cys34). The similar spatial arrangements of the 'back side' of this inhibitor and the 'secondary contact region' of the other inhibitors with respect to the reactive site suggest an important common role of these regions in exhibiting inhibitory activity.

Chemical Phenomena↗

Refined crystal structure (2.3 A) of a double-headed winged bean alpha-chymotrypsin inhibitor and location of its second reactive site.

The crystal structure of a double-headed alpha-chymotrypsin inhibitor, WCI, from winged bean seeds has now been refined at 2.3 A resolution to an R-factor of 18.7% for 9,897 reflections. The crystals belong to the hexagonal space group P6(1)22 with cell parameters a = b = 61.8 A and c = 212.8 A. The final model has a good stereochemistry and a root mean square deviation of 0.011 A and 1.14 degrees from ideality for bond length and bond angles, respectively. A total of 109 ordered solvent molecules were localized in the structure. This improved structure at 2.3 A led to an understanding of the mechanism of inhibition of the protein against alpha-chymotrypsin. An analysis of this higher resolution structure also helped us to predict the location of the second reactive site of the protein, about which no previous biochemical information was available. The inhibitor structure is spherical and has twelve anti-parallel beta-strands with connecting loops arranged in a characteristic beta-trefoil fold common to other homologous serine protease inhibitors in the Kunitz (STI) family as well as to some non homologous functionally unrelated proteins. A wide variation in the surface loop regions is seen in the latter ones.

Binding Sites↗

Marine mammals from northeast Atlantic: relationship between their trophic status as determined by delta13C and delta15N measurements and their trace metal concentrations.

The relationship between trophic position through delta13C and delta15N and trace metal concentrations (Zn, Cd, Cu and Hg) was investigated in the tissues of six marine mammal species from the Northeast Atlantic: striped dolphin Stenella coeruleoalba, common dolphin, Delphinus delphis, Atlantic white-sided dolphin Lagenorhynchus acutus, harbour porpoise Phocoena phocoena, white beaked-dolphin Lagenorhynchus albirostris, grey seal Halichoerus grypus stranded on French Channel and Irish coasts. White-beaked dolphins, harbour porpoises, white-sided dolphins, common and striped dolphins display the same relative and decreasing trophic position, as measured by delta15N values, along both the Irish and French channel coasts, reflecting conservative trophic habits between these two places. Hepatic and renal Cd concentrations were significantly correlated to muscle delta13C and delta15N values while Hg, Zn and Cu did not. These results suggest that Cd accumulation is partly linked to the diet while other factors such as age or body condition might explain Hg, Zn or Cu variability in marine mammals. Combined stable isotope and trace metal analyses appear to be useful tools for the study of marine mammal ecology.

Animals↗

Role of catechol structure in the adsorption and transformation reactions of L-DOPA in soils.

3-(3',4'-Dihydroxyphenyl)-L-alanine (L-DOPA), which is synthesized in velvet bean (Mucuna pruriens), inhibits plant growth. The concentration of L-DOPA in soil is reduced by adsorption and transformation reactions, which can result in the reduction of its plant-growth-inhibitory activity. To determine which part of the L-DOPA structure is involved in the adsorption and soil transformation reactions, we compared the kinetics of L-DOPA disappearance in a volcanic ash soil with that of L-phenylalanine (3-phenyl-L-alanine) and L-tyrosine (3-(4'-hydroxyphenyl)-L-alanine), compounds that are similar in structure to L-DOPA but do not have a catechol (o-dihydroxybenzene) moiety. L-Phenylalanine and L-tyrosine were not adsorbed and transformed in the soil at equilibrium pH values between 4 and 7. These results suggest that the adsorption and transformation reactions of L-DOPA in the soil involve the catechol moiety and not the amino and carboxylic acid groups, which are common to all three compounds. Like L-DOPA, (+)-catechin, another allelochemical that contains a catechol moiety, underwent adsorption and soil transformation reactions. Thus, we concluded that the concentrations of allelochemicals bearing a catechol moiety in soils will decrease rapidly owing to adsorption and transformation reactions, and this decrease will be faster in soils with a high pH value or high adsorption ability. Owing to this decrease in concentration, allelopathic phenomena may not occur.

Adsorption↗

[Hypersensitivity detected by skin tests to food in allergic patients in the Hospital Infantil de México Federico Gómez].

BACKGROUND: The prevalence of food allergy depends on the feeding practices of different parts of the world. Peanuts and nuts cause a great part of allergies in the United States; in Spain and Portugal are commonly due to fish, milk; in Italy seafood and mustard in France. In Mexico we don't know the frequency of food allergy and involved groups of age or specific foods. OBJECTIVE: To determine the frequency of immediate hypersensitivity for foods in allergic children from Hospital Infantil de México Federico Gómez. MATERIAL AND METHODS: A retrospective study was performed between January 1995 to December 1999. The skin prick test (SPT) to food was reviewed according to Aas classification. Male and female from 12 months to 18 years old were reviewed. RESULTS: We documented 1,419 patients with allergy. Of them 442 (31%) had positive skin prick test (SPT) to some of the 33 tested foods. The average age of the study group was of 12.8 years. The most affected was from 4 to 7 years (49%); the second group from 1 to 3 years with 23.7%, and the third was from 12 to 17 years with 13.5%. Fish, milky, seafood, soy, beans, orange, onion, tomato, chicken, nut, lettuce and strawberry, were responsible for 58% of the total of allergic reactions. Of those fish, milk, seafood, soy and orange (39%) had the highest frequency. Hypersensitivity to several foods is low, finding just one allergen in 50% of the cases, two allergens in 25%, three allergens in 9%, four in 7%, five in 5% and more than 6 only in 3% of cases. CONCLUSION: We concluded that almost any kind of food reaction seen in our study was due to alimentary habits of different populations, socioeconomic status, and availability of foods. This indicates the diversity of costumes in Mexican people due to the wide geographical area and cultural background of our country. Results could be increased because of only 33 allergens were tested.

Adolescent↗

[A case-control study on the risk of female breast cancer in Wuhan area].

OBJECTIVE: To investigate the risk factors of female breast cancer and its potential alteration in Wuhan area. METHODS: A case-control study was conducted on 213 cases with histopathological diagnosis and 430 matched controls, using conditional logistic regression analysis. RESULTS: 28 factors such as educational level, history of benign breast disease, age at menarche, age at menopausal, meat and well-done meat intake, soy bean food, fruit, lactation time, body mass index (BMI), juvenile chest X-ray, psychological factor, were associated with breast cancer risk in one-way variance model. Multivariate conditional logistic regression analysis in total significant factors and subgroups showed that the risk factors of breast cancer would include high level education, psychological trauma, history of benign breast disease later age at menopause, more years of menstrual and more years of menstrual before giving first birth, high BMI, well-done meat intake and smoked food. Factors as later menarche, lactate longer, soybean food, fruit, drink tea habit were protective factors for breast cancer. Further breakdown of data showed some difference between premenopausal and postmenopausal women. Risks in premenopausal women were associated with history of benign breast disease, age of menarche, soybean food intake, whereas risks in postmenopausal women were related to age of menopausal, BMI, waist-hip ratio and fruit intake. Both psychological traumatic and duration of lactation were common pre-and postmenopausal risk and protective factors. CONCLUSIONS: Dietary habit and endogenous estrogen exposure related factors played important roles on women breast cancer in Wuhan area.

Adult↗

The conformation of single-stranded nucleic acids tDNA versus tRNA.

Conformational analyses using the single-strand-specific nuclease from mung bean and restriction endonucleases have been performed on a series of DNA fragments related to the sequence of the yeast initiator tRNA(Met). Mung bean nuclease cleaves DNA fragments exclusively in some, but not all, single-stranded regions as predicted by RNA secondary structural rules. Comparison of cleavage patterns of yeast initiator tRNA(Met), tDNA(Met) (a DNA oligomer having the sequence of tRNA(Met] and the anti-tDNA(Met) (the complement of tDNA(Met] suggests that the conformation of the three molecules is very similar. Furthermore, both tDNA and anti-tDNA are cleaved by HhaI and CfoI restriction endonucleases at two GCG/C sites which would be in double-stranded regions (the acceptor and dihydrouridine stem), if the two molecules adopt the tRNA cloverleaf structure. On the other hand, minor cleavage products show that the core region, i.e. the extra loop area, is slightly more exposed in tDNA and in anti-tDNA than in tRNA. Therefore, we submit that the global conformation of nucleic acids is primarily dictated by the interaction of purine and pyrimidine bases with atoms and functional groups common to both RNA and DNA. In this view the 2'-hydroxyl group, in tRNA at least, is an auxiliary structural feature whose role is limited to fostering local interactions, which increase the stability of a given conformation.

Base Sequence↗

Molecular cloning of cDNA, recombinant protein expression and characterization of a buckwheat 16-kDa major allergen.

BACKGROUND: Buckwheat is a common food in Japan, Korea and other countries. A candidate major buckwheat allergen, a 16-kDa protein (BWp16), was previously characterized as a pepsin-resistant protein associated with immediate-type allergies to buckwheat. However, whether recombinant BWp16 can react with a patient's IgE remains uncertain. METHODS: The cDNA encoding BWp16 from Japanese buckwheat seeds was cloned based on the sequences obtained by the 5'-rapid amplification of cDNA ends (RACE) and 3'-RACE PCR. Recombinant BWp16 protein expressed in Escherichia coli was purified using affinity chromatography. Western blotting, ELISA and cross inhibition tests of the purified recombinant BWp16 were performed using sera from patients with positive IgE binding to buckwheat and controls. Pepsin digestion experiments were also performed. RESULTS: The full-length cDNA encodes 149 amino acid residues with a calculated molecular mass of 16.9 kDa. The deduced amino acid sequence included a putative signal peptide sequence. BWp16 showed significant homologies to the buckwheat 8-kDa allergen and Ricinus communis (castor bean) 2S albumin. Sera from patients with positive IgE binding to buckwheat reacted with the purified BWp16. Cross inhibition tests revealed immunological equivalence of the purified recombinant and natural BWp16. The recombinant and natural BWp16 were comparably resistant to pepsin digestion. CONCLUSIONS: BWp16 belongs to the 2S albumin family and is a buckwheat allergen. This purified recombinant BWp16 could be used in the diagnosis of buckwheat allergy.

Allergens↗

Isolation and characterization of the subunits of a heat-labile alpha-amylase inhibitor from Phaseolus vulgaris white kidney bean.

The heat-labile one of the two alpha-amylase inhibitors of the white kidney bean (Phaseolus vulgaris) was found to be composed of three kinds of subunits, and they were isolated and characterized. The alpha-subunit was free from tryptophan and cysteine and the beta-subunit contained no methionine or cysteine. There was no marked resemblance in tryptic peptide maps between the alpha- and beta-subunit polypeptides. The alpha-subunit contained 30% by weight of carbohydrate, mainly made up of high mannose-oligosaccharides, and the sugar moiety of the beta-subunit amounted 7% and appeared to be predominantly composed of xylomannose-type oligosaccharides. The largest subunit, gamma, was very similar in molecular features to a postulated alpha beta-dimer and its N-terminal sequence coincided with that of the alpha-subunit. The molecular weights of the polypeptides of alpha, beta-, and gamma-subunits were shown to be 7,800, 14,000, and 22,000, respectively, by SDS-PAGE. It seemed likely that the alpha- and beta-subunits are common to both of the inhibitors and that the heat-lability of this inhibitor arises from the gamma-subunit.

Amino Acid Sequence↗

Avian sarcoma virus gag and env gene structural protein precursors contain a common amino-terminal sequence.

The initiation site for translation of the avian sarcoma virus glycoprotein precursor, Pr63env, has been determined by analyzing the amino-terminal peptides of Pr63env and the polyprotein precursor Pr76gag encoded by the viral gag gene. The acceptor splice junction used to form the env gene mRNA has also been identified. Hybrid-selected virus-specific mRNAs were translated in vitro in the presence of either L-[35S]methionine to label at every methionine residue or L-[35S]methionine-tRNAMeti to label specifically at the amino-terminal methionine residues. Tryptic peptide maps of Pr63env labeled at every methionine residue contain all of the peptides, plus one additional peptide, present in the map of Pr57env, a nonglycosylated env-encoded polypeptide of molecular weight 57,000 immunoprecipitated from tunicamycin-treated cells. Specific amino-terminal labeling of the in vitro-synthesized polypeptides showed that the peptide missing from Pr57env corresponds to the amino-terminal tryptic peptide of Pr63env, which is removed in vivo as part of the amino-terminal signal peptide. Comparison of the amino-terminal tryptic peptides of Pr63env and Pr76gag showed that they are identical. In contrast, the chymotryptic amino-terminal peptides of Pr76gag and Pr63env are not identical. The location of the acceptor-splice junction in the env mRNA of the Prague A strain of avian sarcoma virus was determined by mung bean nuclease mapping to be at nucleotide 5,078. Fusion of the gag and env gene sequences during splicing results in use of the same AUG codon to initiate synthesis of Pr76gag and Pr63env. This sequence is contained within the 397-nucleotide 5' terminal leader that is spliced to the body of the env mRNA. The possible significance of these results for the regulation of avian sarcoma virus synthesis and translation is discussed.

Amino Acid Sequence↗

Random amplified polymorphic DNA (RAPD) analysis in Indian mung bean (Vigna radiata (L.) Wilczek) cultivars.

Greengram [Vigna radiata (L.) Wilczek], also known as mung bean, widely cultivated in a large number of countries, is an important pulse crop of Asia and is considered one of the ancestral species of the genus Vigna. Since yields of greengram have remained low across subtropical and tropical Asia, it is important to estimate genetic diversity in existing cultivars in order to see if the lack of genetic variability might be a constraining factor. In this study, 32 Indian cultivars of greengram were subjected to random amplified polymorphic DNA (RAPD) analysis using 21 decamer primers. A total of 267 amplification products were formed at an average of 12.71 per primer with an overall polymorphism of 64%. The extent of polymorphism was moderate to low. Jaccard similarity coefficient values ranged from 0.65 to 0.92. The cluster analysis resulted in mainly three clusters revealing greater homology between cultivars released from the same source. The results of principal components analysis also substantiated this conclusion. The close genetic similarity between the cultivars could be explained due to the high degree of commonness in their pedigrees. The narrow genetic base of the greengram cultivars revealed in the present analysis emphasises the need to exploit the large germplasm collections having diverse morphoagronomic traits in cultivar improvement programs.

Crops, Agricultural↗