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PubMed1997

Vacuolar uptake of the phytoalexin medicarpin by the glutathione conjugate pump.

We have studied the uptake of [3H]-medicarpin and its glutathione conjugate(s) into vacuolar membrane vesicles from etiolated hypocotyls of mung bean (Vigna radiata). Unconjugated medicarpin is taken up at a low rate in the presence or absence of MgATP. However, [3H]-medicarpin-glutathione conjugate(s), prepared by incubation of medicarpin with a total maize glutathione S-transferase preparation, is taken up more than four-fold faster than medicarpin in the presence of MgATP, and this uptake is MgATP-dependent. Uptake of medicarpin-glutathione was not significantly inhibited by the ionophore gramicidin-D, but was strongly inhibited by vanadate and the alternative transport substrate S-(2,4-dinitrophenyl) glutathione. Our results demonstrate, in a model system, the potential utilization of the high affinity, high capacity, uncoupler-insensitive glutathione conjugate pump for the vacuolar transport of an isoflavonoid phytoalexin.

Adenosine Triphosphate↗
PubMed2002

Biochemical responses of Cr-tolerant and Cr-sensitive mung bean cultivars grown on varying levels of chromium.

Biochemical investigations were carried out in Cr-tolerant and Cr-sensitive cultivars of mung bean (Vigna radiata L.) with different concentrations of hexavalent chromium (as K2Cr2O7) in hydroponics culture. Seeds were germinated and grown in the presence or absence of chromium under controlled environmental conditions. Protein, pigment and enzyme analysis were conducted in both Cr-tolerant and Cr-sensitive cultivars of mung bean after 72 h of treatments. Chlorophyll and protein contents were reduced in Cr-sensitive cultivars more than those of the tolerant ones. The enzyme activity varied among the Cr-tolerant and Cr-sensitive ones. Activities of catalase, peroxidase, glucose-6-phosphate dehydrogenase and superoxide dismutase were greater in Cr-sensitive than tolerant cultivars.

Adaptation, Physiological↗
PubMed2000

Gahat: a Napalese cure for urolithiasis?

OBJECTIVES: Gahat (Vigna unguiculata) is a legume used for centuries in Nepal and Pakistan to treat the symptoms associated with urinary calculi. We prospectively evaluated the effect of Gahat consumption on 24-hour urine parameters in an attempt to assess its in vivo effect in normal volunteers. METHODS: Eight non-stone-forming volunteers collected 24-hour urine specimens while on their routine diets for baseline data. Urine was analyzed for pH, volume, calcium, citrate, phosphate, sodium, magnesium, uric acid, and oxalate. The Gahat was prepared according to local custom. No additives were used to enhance flavor. The pureed mixture (8 ounces) was ingested three times daily for 2 days. Subjects were instructed to maintain their normal diet, including fluid intake and activity during the study period. Twenty-four hours after the start of Gahat intake, a second 24-hour urine collection was initiated while volunteers continued the Gahat. Results of the urine samples before and after Gahat intake were analyzed, using the paired Student t test. RESULTS: There were no significant differences in urinary electrolytes between the urine samples before and after Gahat intake. Magnesium, urine volume, and uric acid differences approached clinical significance. CONCLUSIONS: Gahat increased urinary magnesium through an unknown mechanism and had no effect on other routine 24-hour urine electrolytes. The increase in urinary volume is attributed to the increase in fluid consumption by the subjects. If this legume is effective in preventing or dissolving urinary calculi, it may act through mechanisms not identified in 24-hour urine electrolytes.

Citrates↗
PubMed1980

Reduction of DL-selenocystine and isolation of L-seleoncysteine.

Cystine, selenocytsine, and several analogs were reduced by dithiothreitol (DTT), beta-mercaptoethanol (ME) and sodium borohydride (NaBH4). DTT was the most effective; DTT to cystine ratios from 10 to 80 were equally effective. With selenocysteine, however, absorption was considerably reduced at all ratios. Selenocysteine was identified as the reduction product by reaction with Gaitonde's reagent, comparison of absorption spectra, paper chromatograhy, utilization by cysteinyl-tRNA synthetase fro Paracoccus denitrificans and Vigna radiata, changes in solubility after DTT treatment, and comparison of infrared spectra. During the ATP-PPi exchange assay, DTT and ME convert cysteine and selenocysteine derivatives to cysteine and selenocysteine which serve as substrates for cysteinyl-tRNA synthetase.

Amino Acyl-tRNA Synthetases↗
PubMed2002

Agroinfection as a rapid method for propagating Cowpea mosaic virus-based constructs.

To increase the efficiency of infections with Cowpea mosaic virus (CPMV)-based constructs, clones suitable for agroinfection were constructed. Full-length copies of RNA-1 and RNA-2 were inserted between the sequence of a Cauliflower mosaic virus (CaMV) 35S promoter and a nos terminator and were introduced into the Agrobacterium tumefaciens plasmid, pBINPLUS. Infiltration of leaves of either Nicotiana benthamiana or cowpea (Vigna unguiculata) with a bacterial suspension containing a mixture of the RNA-1- and RNA-2-based plasmids resulted in the plants developing typical CPMV symptoms. To confirm the utility of this approach for use with CPMV-based vectors, a GFP construct based on RNA-2 was adapted for agroinfection. Infiltration of N. benthamiana leaves with a mixture of Agrobacteria containing this construct and the RNA-1 plasmid resulted in high levels of GFP expression. The results demonstrate that agroinfection is a suitable method for the propagation of CPMV-based derivatives.

Agrobacterium tumefaciens↗
PubMed2003

Effect of air pollution on peri-urban agriculture: a case study.

Peri-urban agriculture is vital for the urban populations of many developing countries. Increases in both industrialization and urbanization, and associated air pollution threaten urban food production and its quality. Six hour mean concentrations were monitored for SO(2), NO(2) and O(3) and plant responses were measured in terms of physiological characteristics, pigment, biomass and yield. Parameter reductions in mung bean (Vigna radiata), palak (Beta vulgaris), wheat (Triticum aestivum) and mustard (Brassica compestris) grown within the urban fringes of Varanasi, India correlated directly with the gaseous pollutants levels. The magnitude of response involved all three gaseous pollutants at peri-urban sites; O(3) had more influence at a rural site. The study concluded that air pollution in Varanasi could negatively influence crop yield.

Air Pollutants↗
PubMed1997

Increased UV-B radiation reduces N2-fixation in tropical leguminous crops.

Net photosynthesis, leaf area, biomass, and number, size and activity of nodules were examined in three leguminous plants subjected under field conditions to supplemental UV-B radiation equivalent to a 15% ozone depletion at 25 degrees N latitude. Enhanced UV-B radiation adversely affected the net photosynthetic rate, growth characteristics and nodule activity in all three species. Maximum reduction in net photosynthesis occurred in Phaseolus mungo cv. Pant U-30, whereas the greatest reduction in nitrogenase activity occurred in Vigna radiata.

Journal Article↗
PubMed1999

Differential effects of cooking in open cups and in sealed cellophane bags on the residual cyanide content of moi-moi, a processed legume product.

Bean pastes from the black eyed bean (Vigna unguiculata), pigeon pea (Cajanus cajan) and Ife brown (V. unguiculata) were processed into a popular West African snack moi-moi, cooked in open aluminium cups and in sealed, thin, transparent cellophane bags. The residual total, free and bound cyanide levels of moi-moi from both processes were determined using a combination of enzymatic and colorimetric procedures. Results showed that in all cases, cooking of moi-moi in open aluminium cups caused retention of significantly higher (P<0.05) residual free cyanide than cooking in sealed cellophane bags. Loss of bound cyanide was unaffected by cooking procedure. These results suggest that cooking of moi-moi in cellophane bags reduces the risks of cyanide exposure from legumes.

Cellophane↗
PubMed1997

Theophylline metabolism in higher plants.

Metabolism of [8-(14)C]theophylline was investigated in leaf segments from Camellia sinensis (tea), Camellia irrawadiensis, Ilex paraguariensis (maté) and Avena sativa, root segments of Vigna mungo seedlings and cell suspension cultures of Catharanthus roseus. There was extensive uptake and metabolism of [8-(14)C]theophylline by leaves of tea and Camellia irrawadiensis and, to a lesser extent, maté. These purine alkaloid-containing species converted [8-(14)C]theophylline into 3-methylxanthine, xanthine, the ureides allantoin and allantoic acid, and CO2. With the other test systems, which were from species that do not produce purine alkaloids, there were low levels of [8-(14)C]theophylline uptake which were accompanied by incorporation of relatively small amounts of label into 3-methylxanthine, xanthine and CO2. None of the higher plants converted [8-(14)C]theophylline to either 1-methyluric acid or 1,3-dimethyluric acid, which are the main catabolites of theophylline in mammals. The data indicate that the main route of theophylline degradation in higher plants involves a theophylline --> 3-methylxanthine --> xanthine --> uric acid --> allantoin --> allantoic acid --> --> CO2 + NH3 pathway. In tea and mate, large amounts of [8-(14)C]theophylline were also converted to theobromine and caffeine via a theophylline --> 3-methylxanthine --> theobromine --> caffeine salvage pathway. The diversity of theophylline metabolism in higher plants and mammals is discussed.

Plants↗
PubMed1998

Protein stability in the amorphous carbohydrate matrix: relevance to anhydrobiosis.

The formation of intracellular glass is proposed to be relevant to protein stabilization and survival of anhydrobiotic organisms in the dry state. The stability of proteins in the amorphous carbohydrate matrix and its relevance to seed survival have been investigated in the present study. Glucose-6-phosphate dehydrogenase (G6PDH) was preserved in the amorphous glucose/sucrose (1:10, w/w) matrix by freeze-drying. The stability of freeze-dried G6PDH was examined at temperatures above and below the glass transition temperature (Tg). The rate of G6PDH inactivation in the amorphous carbohydrate matrix deviated significantly from the Arrhenius kinetics, and conformed to the Williams-Landel-Ferry (WLF) relationship. The temperature dependence of G6PDH inactivation in two sets of samples with different Tg values was compared. Identical temperature dependence of G6PDH inactivation was observed after temperature normalization by (T-Tg). Seed survival of Vigna radiata Wilczek (mung bean) showed a similar WLF kinetics at storage temperatures T > or = Tg. In situ protein stability in mung bean embryonic axes was studied using differential scanning calorimetry (DSC). Thermal stability of seed proteins exhibited a strong dependence on the Tg of intracellular glass. These results indicate an important role of the glassy state in protein stabilization. Our data suggest an association between protein stability in intracellular glass and seed survival during storage.

Calorimetry, Differential Scanning↗
PubMed2000

Divergent Bradyrhizobium symbionts on Tachigali versicolor from Barro Colorado Island, Panama.

Relationships of root-nodule bacteria from the tree Tachigali versicolor (legume subfamily Caesalpinioideae) were analyzed for 20 isolates sampled from juvenile plants growing on Barro Colorado Island (BCI), Panama. Bacterial genetic diversity appeared to be low. In the highly polymorphic 5' intervening sequence region of 23S rRNA, all isolates had the same length variant. A 472 bp segment spanning this region was sequenced in four isolates, and all proved to be identical at every nucleotide position. RFLP analysis of a 868 bp fragment of the nitrogenase alpha-subunit gene likewise indicated that all 20 isolates shared an identical set of restriction sites. Phylogenetic analysis of both partial 23S rRNA and nearly full-length 16S rRNA sequences showed that bacterial symbionts of T. versicolor fall into the genus Bradyrhizobium. However, they are divergent from the bradyrhizobia associated with other BCI legumes, as well as from other currently known bacteria in this genus. Inoculation tests with two promiscuously-nodulating legumes showed that bacteria from T. versicolor were unable to form nodules on Vigna unguiculata, but did nodulate Macroptilium atropurpureum, although the nodules lacked nitrogen fixation activity. The association of Tachigali with a divergent lineage of Bradyrhizobium is noteworthy in view of this plant's position within a clade of the mostly non-nodulating "primitive" legume subfamily Caesalpinioideae that gave rise to the predominantly nodulating subfamily Mimosoideae.

Base Sequence↗
PubMed1987

Physical exercise and the neuroendocrine control of reproduction.

Reproductive change during conditioning exercise (physical training provides a model of hypothalamic adaptation to alterations in the external and internal environment. Parallels exist between the reproductive changes with exercise and those occurring with physical illness, undernutrition and psychological trauma. Although menstrual cyclicity may be disrupted in younger women, luteal phase shortening, anovulation and decreased premenstrual symptoms within normal ovulatory cycles are the most frequent observations noted. Baseline LH, prolactin and oestradiol tend to be lower, and other hormones unchanged, in trained women. Testosterone may be decreased within the normal range in men. Recent evidence shows that LH pulse frequency, amplitude and area under the LH curve are decreased in both female and male runners. Interrelationships between increases in central dopamine, endorphin and probably some hypothalamic message(s) relating to nutritional state appear to modulate these reproductive changes. The clinical and therapeutic response to reproductive alterations in the context of exercise differs when these are seen as adaptive and not as disease processes (Prior and Vigna, 1985b).

Energy Metabolism↗
PubMed2001

Digestion of legume starch granules by larvae of Zabrotes subfasciatus (Coleoptera: bruchidae) and the induction of alpha-amylases in response to different diets.

Zabrotes subfasciatus larvae possess three alpha-amylase isoforms as determined by in gel assays following SDS-PAGE. The two minor isoforms present lower electrophoretic mobility than the major form, and seem to occur as a heterodimer. When developed inside Vigna unguiculata (cowpea) seeds, fourth instar larvae have minor quantities of the slow-migrating forms, but when reared on seeds of Phaseolus vulgaris (common bean) or Phaseolus lunatus, the two slow-migrating forms are expressed in higher amounts, while activity of the major form was independent of the host seed. Larvae developing inside cowpea seeds at the beginning of the fourth instar were fed on flour from cotyledons of cowpea or common bean. Larvae fed on the common bean flour started to express the dimer in higher amounts when compared with the control larvae fed on cowpea flour. In an attempt to correlate differences between starch granules and the induction of alpha-amylases, a detailed study on the digestive process of the granules was conducted. Incorporation of purified starch granules into artificial diets did not induce the two minor alpha-amylases. The in vitro hydrolysis rates of purified granules and the pattern of dextrins liberated by the different alpha-amylases were similar for the two legume species. The starch granules enter the midgut extensively damaged, which may facilitate the access to the more susceptible parts of the granules to enzymatic attack.

Animals↗
PubMed2002

Comparison of soxhlet and microwave-assisted extractions for the determination of fenitrothion residues in beans.

White and black "niebe" beans [Vigna unguiculata (L.) Walp] from Senegal were treated with fenitrothion (O,O-dimethyl O-4-nitro-m-tolyl phosphorothioate), and the residues were determined by high-performance liquid chromatography (HPLC) and electron capture gas chromatography (EC-GC). Fenitrothion residues from the beans were extracted by Soxhlet extraction (SE) and microwave-assisted extraction (MAE). A column cleanup procedure was used to remove the coextractives in the extract before HPLC and EC-GC analyses. The overall mean recoveries of fenitrothion residues in the 0.19-1.90 microg/kg fortification range determined from extracts obtained by SE and MAE were 88.4 and 89.8%, respectively, with respective relative standard deviations of <4%. The results show that MAE is a viable alternative to the commonly used SE for the determination of fenitrothion residues in beans.

Chromatography, Gas↗
PubMed2003

Evaluation of the estrogenic effects of legume extracts containing phytoestrogens.

Seven legume extracts containing phytoestrogens were analyzed for estrogenic activity. Methanol extracts were prepared from soybean (Glycine max L.), green bean (Phaseolus vulgaris L.), alfalfa sprout (Medicago sativa L.), mung bean sprout (Vigna radiata L.), kudzu root (Pueraria lobata L.), and red clover blossom and red clover sprout (Trifolium pratense L.). Extracts of kudzu root and red clover blossom showed significant competitive binding to estrogen receptor beta (ERbeta). Estrogenic activity was determined using an estrogen-dependent MCF-7 breast cancer cell proliferation assay. Kudzu root, red clover blossom and sprout, mung bean sprout, and alfalfa sprout extracts displayed increased cell proliferation above levels observed with estradiol. The pure estrogen antagonist, ICI 182,780, suppressed cell proliferation induced by the extracts, suggesting an ER-related signaling pathway was involved. The ER subtype-selective activities of legume extracts were examined using transiently transfected human embryonic kidney (HEK 293) cells. All seven of the extracts exhibited preferential agonist activity toward ERbeta. Using HPLC to collect fractions and MCF-7 cell proliferation, the active components in kudzu root extract were determined to be the isoflavones puerarin, daidzin, genistin, daidzein, and genistein. These results show that several legumes are a source of phytoestrogens with high levels of estrogenic activity.

Breast Neoplasms↗
PubMed2003

Nutritional and physiological responses of young growing rats to diets containing raw cowpea seed meal, protein isolate (globulins), or starch.

The nutritional and physiological effects of raw cowpea (Vigna unguiculata (L) Walp.) seed meal, protein isolate (globulins), or starch on the metabolism of young growing rats have been evaluated in 14-day trials. Wet and dry weight gain, feed conversion efficiency, and lipid and protein accretion were significantly reduced as a result of inclusion of seed meal, globulins, or starch in the diet, with growth retardation being most marked with the seed meal. The proportional weights of the small intestine and pancreas were increased by meal diets, and serum cholesterol levels were slightly reduced. The globulins and raw starch also increased relative small intestine weights but had no effect on the pancreas or serum constituents. The effects of cowpeas on rats appeared to be due primarily to the combined actions of globulins, resistant starches, protease inhibitors, and possibly fiber and non-starch polysaccharides on intestinal and systemic metabolism.

Animal Nutritional Physiological Phenomena↗
PubMed2004

A Kunitz-type inhibitor of coleopteran proteases, isolated from Adenanthera pavonina L. seeds and its effect on Callosobruchus maculatus.

The cowpea weevil Callosobruchus maculatus is one of the major pests of Vigna unguiculata cowpea. Digestion in the cowpea weevil is facilitated by high levels of cysteine and aspartic acid proteinases. Plants synthesize a variety of molecules, including proteinaceous proteinase inhibitors, to defend themselves against attack by insects. In this work, a trypsin inhibitor (ApTI) isolated from Adenanthera pavonina seeds showed activity against papain. The inhibition of papain by ApTI was of the noncompetitive type, with a K(i) of 1 microM. ApTI was highly effective against digestive proteinases from C. maculatus, Acanthoscelides obtectus (bean weevil), and Zabrotes subfasciatus (Mexican bean weevil) and was moderately active against midgut proteinases from the boll weevil Anthonomus grandis and the mealworm Tenebrio molitor. In C. maculates fed an artificial diet containing 0.25% and 0.5% ApTI (w/w), the latter concentration caused 50% mortality and reduced larval weight gain by approximately 40%. The action of ApTI on C. maculatus larvae may involve the inhibition of ApTI-sensitive cysteine proteinases and binding to chitin components of the peritrophic membrane (or equivalent structures) in the weevil midgut.

Animals↗
PubMed2005

Inositol phosphate profiling of fermented cowpeas by 1H NMR spectroscopy.

The inositol phosphate content of naturally fermented cowpeas (Vigna sinensis var. Carilla) was studied using ion-pair HPLC and 1H NMR spectroscopy. The fermented flour was extracted with 0.5 M HCl, and the extract was purified and fractionated by ion-exchange chromatography. 1H NMR allowed for the identification of two monophosphates [Ins(1 or 3)P1 and Ins(4 or 6)P1], one inositol diphosphate [Ins(1,4)P2], three inositol triphosphates [Ins(1,2,6)P3, Ins(1,5,6)P3, and Ins(1,4,5)P3], one inositol tetraphosphate [Ins(1,3,4,5)P4], and one inositol pentaphosphate [Ins(1,2,3,5,6)P5]. Some of these isomers [Ins(1,4,5)P3 and Ins(1,3,4,5)P4] are considered to play important biological roles in intracellular signaling.

Chromatography, High Pressure Liquid↗