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A new biochemical marker for aluminium tolerance in plants.

Al was shown to elicit the induction of several pathogenesis-related genes, suggesting that a common signalling pathway may be involved in the early response to Al and pathogens. However, we found no evidence of oxidative burst involving either H2O2 or O2- during the first hours of Al exposure distinguishing the early response to Al from a common response to pathogen infection. We identified a strong superoxide dismutase insensitive nitro blue tetrazolium (NBT) reduction activity in the root tips of control plants. This activity was rapidly inhibited by Al exposure in the meristematic/distal transition zones of roots in all species examined. In wheat (Triticum aestivum), the inhibition of NBT reduction occurred in less than 1 min in vivo suggesting that Al either directly blocks an enzyme responsible for NBT reduction, or affects a signal pathway involved in the regulation of this activity. The sensitivity of NBT reduction to KCN and NaN3 suggests that an enzymatic, rather than a chemical reaction is involved. In tolerant plants, the inhibition of NBT reduction caused by Al was reversed within 24 h of exposure. The level of recovery was a function of the degree of Al tolerance. We show that NBT reduction is a simple biochemical marker allowing the rapid identification of tolerant individuals within a segregating population.

Journal Article↗

Effect of potato on acid-base and mineral homeostasis in rats fed a high-sodium chloride diet.

Excessive dietary NaCl in association with a paucity of plant foods, major sources of K alkaline salts, is a common feature in Western eating habits which may lead to acid-base disorders and to Ca and Mg wasting. In this context, to evaluate the effects of potato, rich in potassium citrate, on acid-base homeostasis and mineral retention, Wistar rats were fed wheat starch (WS) or cooked potato (CP) diets with a low (0.5 %) or a high (2 %) NaCl content during 3 weeks. The replacement of WS by CP in the diets resulted in a significant urinary alkalinisation (pH from 5.5 to 7.3) parallel to a rise in citrate and K excretion. Urinary Ca and Mg elimination represented respectively 17 and 62 % of the daily absorbed mineral in rats fed the high-salt WS diet compared with 5 and 28 % in rats fed the high-salt CP diet. The total SCFA concentration in the caecum was 3-fold higher in rats fed the CP diets compared with rats fed the WS diets, and it led to a significant rise in Ca and Mg intestinal absorption (Ca from 39 to 56 %; Mg from 37 to 60 %). The present model of low-grade metabolic acidosis indicates that CP may be effective in alkalinising urine, enhancing citrate excretion and ameliorating Ca and Mg balance.

Acid-Base Equilibrium↗

Bread fortified with cholecalciferol increases the serum 25-hydroxyvitamin D concentration in women as effectively as a cholecalciferol supplement.

Fortification of foods is a feasible way of preventing low vitamin D status. Bread could be a suitable vehicle for fortification because it is a common part of diets worldwide. The bioavailability of cholecalciferol from bread is not known. We studied cholecalciferol stability, the concentration of the added cholecalciferol, the dispersion of cholecalciferol in bread, and the bioavailability of cholecalciferol from fortified bread. Three batches of fortified low-fiber wheat and high-fiber rye breads were baked; from each batch, 3 samples of dough and bread were analyzed for their cholecalciferol content. In a single-blind bioavailability study, 41 healthy women, 25-45 y old, with mean serum 25-hydroxyvitamin D concentration 29 nmol/L (range 12-45 nmol/L), were randomly assigned to 4 study groups. Each group consumed fortified wheat bread, fortified rye bread, regular wheat bread (control), or regular wheat bread and a cholecalciferol supplement (vitamin D control) daily for 3 wk. The daily dose of vitamin D was 10 mug in all groups except the control group. The vitamin dispersed evenly in the breads and was stable. Both fortified breads increased serum 25-hydroxyvitamin D concentration as effectively as the cholecalciferol supplement. Supplementation or fortification did not affect serum intact parathyroid hormone concentration or urinary calcium excretion. In conclusion, fortified bread is a safe and feasible way to improve vitamin D nutrition.

Adult↗

Infectivity and complete nucleotide sequence of the genome of a South African isolate of maize streak virus.

The complete infectious genome of a South African isolate of the geminivirus maize streak (MSV-S) has been cloned, characterized, and sequenced. Using an A. tumefaciens Ti plasmid delivery system, the cloned -2.7 kb single circular MSV component was shown to be necessary and sufficient for infection of maize. Based on sequence analysis of the infectious clone, MSV-S is highly homologous to the previously characterized Kenyan and Nigerian isolates. While the genomic organization of MSV-S has elements in common with each of these previously characterized isolates, it is identical to neither and its analysis addresses the discrepancies between them. The result is a somewhat simplified and unified picture of the viral genome, the structural organization of which is essentially identical to that of wheat dwarf virus.

Base Sequence↗

Localization of the binding sites of prokaryotic and eukaryotic RNA polymerases on simian virus 40 DNA.

The binding sites of calf thymus RNA polymerase (B) II, wheat germ RNA polymerase B and of the Escherichia coli RNA polymerase were mapped on the simian virus 40 genome by observation of enzyme-linear DNA complexes by electron microscopy. Three to four major sites and several minor sites are observed for each enzyme; common binding sites for the three enzymes are found in positions 0.17, 0.53 and 0.90 of the viral physical map. Initiation complexes with these enzymes can be stabilized with specific ribodinucleotides and a single ribonucleoside triphosphate. Whereas ApA and ATP greatly enhances the binding of the E. coli enzyme at position 0.17, they stabilize the binding of the eukaryotic enzyme at many sites, some of them located in close proximity of the origin of replication.

Adenosine Monophosphate↗

Coexistence of related pathogen species on arable crops in space and time.

This review considers factors affecting the coexistence of closely related pathogen species on arable crops, with particular reference to data available at Rothamsted for Septoria tritici/Stagonospora nodorum (Mycosphaerella graminicola/Phaeosphaeria nodorum) (septoria leaf blotch diseases on winter wheat), Oculimacula yallundae/O. acuformis (eyespot disease of winter cereals), and Leptosphaeria maculans/L. biglobosa (phoma stem canker on winter oilseed rape). Factors affecting the short-term, medium-term, and long-term coexistence of such related pathogen species are reviewed, and their evolution from common ancestors considered. Small niche differences between the related pathogen species enable them to coexist on the same host. The niche differences result from small differences in their biology/epidemiology, leading to separation in space, time, or resource use. Changes in both natural (e.g., fluctuating temperature) and man-made (e.g., agronomic practices, pollution) factors influence the coexistence. Such factors may result in coexistence between the related species in some parts of the world, whereas in other parts only one species occurs. These principles illustrated with pathogens of arable crops are generic to other host-pathogen systems.

Biological Evolution↗

A novel PCR method for quantification of buckwheat by using a unique internal standard material.

A novel quantitative and specific method for detection of buckwheat, a known food allergen, in diverse food materials was developed by using a unique internal standard to compensate for the variability in DNA extraction and amplification efficiencies. The method was based on a real-time PCR targeting the internal transcribed spacer region of Fagopyrum spp. and was designed to detect both cultivated and wild buckwheat, because wild buckwheat might be potentially allergenic. As the internal standard material, ground seeds of statice (Limonium sinuatum) were added to food samples prior to DNA extraction, and the amount of statice DNA measured by real-time PCR was used to standardize the buckwheat content. Statice, an ornamental plant, was chosen as the internal standard material because it was readily available and was inferred to be least likely to be commingled in foods. The specificity of the PCR system was tested against commonly used food materials of plant origin. Quantitative results expressed in buckwheat protein concentrations (mean +/- standard deviation) for various food samples prepared to contain 10 ppm (wt/wt) of buckwheat flour (corresponding to 1.2-microg/g [ppm] buckwheat protein) ranged from 0.7 +/- 0.2 (rice) to 0.9 +/- 0.4 (wheat) and for 100-ppm (wt/wt) samples (12-microg/g [ppm] buckwheat protein) from 7.7 +/- 1.0 (pepper) to 9.8 +/- 0.5 (wheat) microg/g (ppm). The method's accuracy, sensitivity, and specificity were considered sufficient for detection of buckwheat contamination at the level required for compliance with the Japanese Food Allergen Labeling Regulation.

Allergens↗

[Serum antibodies in dogs with autoimmune disorders recognize 40 kd glycoprotein in the nucleus of mammalian cells].

We report a new antibody specificity in 15 sera recovered from a group of dogs developing systemic lupus erythematosus (SLE) or clinically related disorders. This antibody stains in a speckled fashion the nucleus of human Hep-2 cells. Immunodiffusion tests with saline extracts of rabbit thymus showed that all 15 sera generate a common precipitation line which crosses the lines from reference sera to Sm, SS-A/ro, SS-B/La, and RNP antigens. The target nuclear antigen is a 40 kD polypeptide (p40). An important property of p40 resides in its ability to bind specifically Wheat Germ Agglutinin lectin but not Concanavalin A, supporting the notion that the antigen is a glycoprotein bearing a N-acetylglucosamine moiety.

Animals↗

Bioactive phytochemicals with emphasis on dietary practices.

Diet can modify the pathophysiological processes of various metabolic disorders and can be an effective preventive strategy for various disease processes most of which are known to involve oxidative damage. Both nutrient and non-nutrient components of the diet have been recognized for their anti-oxidant and other potential benefits. Plant foods contain phytochemicals such as flavonoids, phenolic acids, etc., which show biological activity. Some common foods used in Indian culinary practices were assessed for their anti-oxidant, anti-mutagenic and anti-carcinogenic effects and vitamin D activity and evaluated for their plausible biological effects. Green leafy vegetables had the highest anti-oxidant activity followed by wheat and rice. Cooking decreased this activity. Eugenol, the active principle of clove, was shown to offer protection against CCl4 induced hepatotoxicity in rats. It also showed anti-peroxidative activity in addition to decrease in O2 formation. Studies on the anti-carcinogenic effect of turmeric/curcumin revealed that both are potent anti-mutagens in vivo and reduce the adducted DNA levels in liver of rates challenged with B(a)P. In another study, Syrian hamsters receiving turmeric/curcumin through diet or local paint on cheek pouch had lower tumour burden as well as adducted DNA level against 7-12-DMBA challenge. Turmeric/curcumin were found to be better anti-tumour agents when given in the post initiation phase of carcinogenesis. The beneficial effect of turmeric was found to be due to its anti-oxidant potential. Studies on humans at risk of palatal cancer due to reverse smoking showed that turmeric (1 g/day) for 9 months had a significant impact on the regression of precancerous lesions. Onion and garlic also possess anti-mutagenic principle. Further studies on the bioactive phytochemicals in plants showed that certain plants belonging to Solanaceae (Cestrum diurnum, Lycopersicon esculentum and Solanum melongena) have calcinogenic potential and vitamin D like activity. In view of the vast data on bioactive principles from plants, it is suggested that dietary prevention coupled with other life-style changes in perhaps the right answer for prevention of cancer and other chronic diseases in India.

Animals↗

Stimulation of colony formation and growth factor production of human T lymphocytes by wheat germ lectin.

Wheat germ lectin (WGL) induced proliferation of T-lymphocyte colony-forming units (TL-CFU) from human peripheral blood mononuclear cells (PBMC) in a one stage agar culture needing higher cell numbers than phytohaemagglutinin (PHA). However, WGL preactivated TL-CFU in a first stage liquid culture for the growth of PHA-dependent T-lymphocyte colonies in a subsequent agar culture. These results suggested that WGL and PHA have biological activities in common. WGL and PHA both induced the expression of Interleukin 2 (IL-2) receptors by PBMC to become responsive to IL-2, although production of IL-2 by WGL-stimulated PBMC was low compared with PHA-stimulated PBMC. This might account for the low proliferative response of PBMC to WGL.

Agar↗

The dependence of the in vitro fermentation of dietary fibre to short-chain fatty acids on the contents of soluble non-starch polysaccharides.

The fermentability of cellulose and dietary fibre in common clinical use (Inolaxol, Fiberform, Vi-Siblin, Lunelax, pectin) was measured as the in vitro production of short-chain fatty acids, lactate, and ammonia in 16.6% faecal homogenates from 18 healthy volunteers. The results were compared with the contents of soluble and insoluble non-starch polysaccharides as determined by the method of Englyst. The amounts of soluble non-starch polysaccharides in the fibre were closely associated with the mean productions of short-chain fatty acids after 6 h (R = 0.94, p < 0.002) and 24 h (R = 0.98, p < 0.0002) of incubation. The mean production of ammonia was inversely related to the soluble fraction of the fibre (after 6 h, R = -0.93, p < 0.003; after 24 h, R = -0.90, p < 0.006). These variables were not dependent on the insoluble fractions of the fibre. The in vitro fermentability differed considerably among the fibres: cellulose and Inolaxol (sterculia gum) were almost non-fermentable, Fiberform (wheat bran-based) was low-grade fermentable, Vi-Siblin and Lunelax (both ispaghula husk) were intermediately fermentable, and pectin was highly fermentable. These findings support that the water solubility determines the degree of fermentability of dietary fibre and thereby the corresponding bacterial assimilation of ammonia. In vitro measurements of short-chain fatty acid production in faecal homogenates may hence supplement commonly used methods to classify dietary fibre.

Adult↗

Isolation, identification, and accumulation of 2-acetamidophenol in liquid cultures of the wheat take-all biocontrol agent Pseudomonas fluorescens 2-79.

Pseudomonas fluorescens strain 2-79 (NRRL B-15132) is a classic biological control agent known to produce phenazine-l-carboxylic acid (PCA) as its primary means of suppressing take-all disease of wheat. In addition to PCA, an unknown metabolite was discovered in a liquid culture used to produce the biocontrol agent. The objective of the current study was to isolate, identify, and evaluate the accumulation of this compound in production cultures. Upon centrifugal fractionation of a production culture, thin-layer chromatography (TLC) analyses of extracts of the cells and cell-free supernatant indicated the compound to be primarily in the supernatant. Purified compound was obtained by extraction of culture supernatant, followed by flash chromatography of the extract and preparative TLC. The 'H and 13C nuclear magnetic resonance and electron impact mass spectra indicated the compound to be 2-acetamidophenol (AAP). Measured by reversed-phase HPLC, the accumulations of AAP and PCA in cultures of strain 2-79 reached 0.05 g/l and 1 g/l, respectively. The accumulations of AAP and PCA in liquid cultures were linearly correlated (P < 0.001), as shown by studies of cultures stimulated to yield varying levels of PCA by controlling levels of oxygen transfer, pH, and growth medium composition. In this study, oxygen limitation, a defined amino-acid-free medium, and neutral pH stimulated maximal production of both AAP and PCA. Furthermore, a transposon mutant of 2-79 [2A40 2-79 (phz-)] unable to produce PCA did not accumulate AAP. These findings indicate that AAP and PCA are likely to share a common segment of biosynthetic pathway. This is the first report of AAP production by a strain of P. fluorescens. Possible routes of AAP production are discussed relative to current knowledge of the phenazine biosynthetic pathway of strain 2-79. The pertinence of AAP to the design of commercial seed inoculants of phenazine-producing bacteria for controlling wheat take-all is also considered.

Acetaminophen↗

In vitro transcription and translation of cloned cDNAs encoding the 30-kDa protein gene of TMV.

A cDNA clone encoding the nonstructural, 30-kDa protein of the common (U1) strain of tobacco mosaic virus (TMV) was isolated and characterized. cDNA clones representing the intact gene as well as deletions from the 5' end of the gene were subcloned into SP6 vectors. Capped RNAs produced by in vitro transcription reactions were translated in a wheat germ cell-free system. The resultant proteins were compared to proteins obtained from the in vitro translation of intermediate length (I2) rods of TMV. Transcripts of the cDNA clones encoded polypeptides of 30, 28, or 18 kDa that were immunoprecipitated by antibody prepared against a synthetic peptide representing the carboxy terminus of the 30-kDa protein. cDNA clones containing the intact 30-kDa sequence coded for 30-kDa polypeptides while clones lacking the 30-kDa initiation codon produced 28-kDa polypeptides. Surprisingly, translation of a transcript from a cDNA clone containing the 30-kDa gene plus 390 nucleotides 5' of the initiator AUG yielded a polypeptide with an approximate molecular mass of 18 kDa. The results indicate that an intact and functional 30-kDa protein gene has been cloned. The significance of these results, with respect to determining the function of the 30-kDa protein, is discussed.

Cell-Free System↗

Diet and colorectal cancer: current evidence for etiology and prevention.

The etiology of colorectal cancer (CRC) involves the interaction of cell molecular changes and environmental factors, with a great emphasis on diet components. But the paths connecting lifestyle characteristicas and the colorectal carcinogenesis remain unclear. Several risk factors are commonly found in western diets, such as high concentrations of fat and animal protein, as well as low amounts of fiber, fruits and vegetables. A large number of experimental studies have found a counteractive effect of fiber on neoplasia induction, especially in relation to fermentable fiber (wheat bran and cellulose). Epidemiological correlation studies have also indicated that a greater ingestion of vegetables, fruit, cereal and seeds is associated to a lower risk for colorectal neoplasia. Moreover, beneficial properties of fiber (especially from vegetable sources) were documented in more than half of case-control studies. Nevertheless, recent epidemiological data from longitudinal and randomized trials tended not to support this influence. Future research should evaluate what sources of fiber provide effective anti-neoplasic protection, carrying out interventional studies with specific fibers for longer periods. Red meat, processed meats, and perhaps refines carbohydrates are also implicated in CRC risk. Recommendantions to decrease red meat intake are well accepted, although the total amount and composition of specific fatty acids may have distinct roles in this setting. Current evidence favors the substitution of long and medium-chain fatty acids and arachidonic acid for short-chain fatty acids and eicosapentaenoic acid. Excess boy weight and excess energy intake inducing hyperinsulinemia have been also associated to CRC, as well as personal habits such as physical inactivy, high alcohol consumption, smoking and low consumption of folate and methionine. Thus, current recommendations for decreasing the risk of CRC include dietary measures such as increased plant food intake; the consumption of whole grains, vegetables and fruits; and reduced red meat intake.

Colorectal Neoplasms↗

Lectin binding sites in developing mouse limb buds.

The binding sites of the following biotinylated lectins were demonstrated in serial paraffin sections of fore- and hindlimb buds from day-9 to day-16 mouse embryos with the Avidin-Biotin-Peroxidase Complex (ABC) procedure: Concanavalin A (Con A), Soybean Agglutinin (SBA), Wheat Germ Agglutinin (WGA), Peanut Agglutinin (PNA), Ricinus Communis Agglutinin I (RCA), Ulex Europaeus I Agglutinin (UEA), and Dolichos Biflorus Agglutinin (DBA). Alternating neighbouring sections were used to compare the distribution of PNA staining, PNA staining after neuraminidase treatment (N-PNA) and the autoradiographic sites of [35S]-sulphate uptake. Unspecific binding sites common to all lectins tested were observed in periderm and chondrocytes. Several lectin affinities were seen in the undifferentiated mesoderm (Con A, WGA, RCA), blood vessels (WGA, PNA, N-PNA, RCA, UEA, DBA) and macrophages (Con A, WGA, N-PNA, RCA). A very selective and mainly extracellular affinity to N-PNA was demonstrated in the condensed preskeletal mesoderm, where it characterizes indistinct prospective chondrogenic, perichondral and pre-articular areas. Comparison with the distribution pattern of [35S]-sulphate uptake and other previously published histochemical data suggests that N-PNA staining occurs at the late blastema stage, i.e. after the stage of cell condensation and before the earliest deposit of stainable matrix in chondrogenic areas. This property later disappears from the chondrifying rudiments, and is maintained in perichondral and pre-articular tissues. Surprisingly, only the pre-articular areas bind PNA without pretreatment with neuraminidase. A transient RCA binding probably related to terminal morphogenesis was detected in the undifferentiated distal part of the predigital columns of day-12 and day-13 limb buds. From the day-13 stage onwards, diverse new lectin affinities appeared in differentiating tissues, such as pretendinous rudiments, perichondrium and prospective periosteum, muscular connective tissue, myotubes, superficial fasciae and prospective dermis. A strong SBA and PNA staining was also detected in the extracellular matrix associated with the epithelial septa separating the roots of the digits in day-15 and day-16 limb buds.

Animals↗

T3-T cell receptor (Ti) complex-independent activation of T cells by wheat germ agglutinin.

The T3-Ti complex appears to play a central role in the activation of T cells by antigens and mitogens. Wheat germ agglutinin (WGA) is a unique lectin which inhibits T cell proliferation induced by mitogens, but it also induces marked IL 2 production by peripheral blood T cells. The pattern of responses induced by WGA suggests that this lectin may use a different mechanism of T cell activation other than the mechanism employed by the common T cell stimulants. We first investigated the production of IL 2 by Jurkat cells (E6-1) stimulated with WGA, before and after modulation of the surface T3-Ti complex. IL 2 production was markedly reduced after modulation of the T3 antigen from the cell surface when these cells were stimulated with PHA. In contrast, little change was observed in WGA-induced IL 2 production after modulation. Furthermore, we examined the effect of WGA on a T3-mutant of E6-1 cells (T3.1) which does not produce IL 2 in response to PHA or PHA plus PMA. WGA-stimulated T3.1 cells produced a significant amount of IL 2 with or without added PMA. In addition, a small but consistent rise in intracytoplasmic free calcium was observed when these cells were stimulated with WGA. These results demonstrate the presence of an alternative mechanism of T cell activation independent of the T3-Ti complex.

Acetylglucosamine↗

Characterization and genomic organization of Ty1-copia group retrotransposons in rye (Secale cereale).

The genomic organisation of the Ty1-copia retrotransposons in rye (Secale cereale) has been studied. We have used the polymerase chain reaction (PCR) to amplify sequences from a conserved domain of the reverse transcriptase gene of the Ty1-copia retrotransposons in this species. Sequence analysis of 26 of these PCR products shows them to be a highly heterogeneous population, a feature that is common in plants. Slot blot analysis shows that there are about 100,000 individual Ty1-copia retrotransposons in rye. In situ hybridization of a heterogeneous probe, representing the whole population of rye Ty1-copia retrotransposon sequences, to chromosome spreads of triticale (xTriticosecale), a rye-wheat hybrid, shows that these sequences are present throughout all the rye chromosomes but absent from the centromeric regions and, in particular, from the terminal heterochromatin. Southern analysis of oat, barley, wheat, and rye, using as a probe R9, one of the rye sequences that is closely similar to the BARE-1 element of barley, shows that close relatives of this retrotransposon subgroup are present in all these species in high copy number. Northern analysis on RNAs from seedlings shows that the BARE-1 subgroup is transcribed in all these cereal plants but in variable amounts: high in barley, moderate in wheat and rye, and extremely low in oat.

Amino Acid Sequence↗

The impact of household preparations on the residues of pesticides in selected agricultural food commodities available in India.

Gas chromatographic multiresidue methods for simultaneous determination of organophosphorus, organochlorine, and organonitrogen pesticides were used to study the exposure of the Indian population to pesticide contamination at their actual dietary intakes. Selected agricultural commodities--5 kinds of vegetables (tomato, potato, okra, cabbage, and green beans), 6 kinds of cereals and pulses (rice, maize, wheat, red gram, black gram, and green gram), and 6 kinds of fruits (mango, orange, guava, banana, apple, and grapes)--readily available in Chennai City local markets--were studied for this purpose. Samples were fortified with known concentrations of various pesticides and subjected to household preparation methods commonly used in India. The impact of household preparation is very high, resulting in 65-95% decontamination of pesticides at different stages. Of 512 raw market samples analyzed, the organochlorine and organophosphorus pesticides present in 12 samples were removed during household preparations, resulting in residues well below the toxicologically acceptable limits.

Chromatography, Gas↗