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Effects of crude extracts of Mucuna pruriens (Fabaceae) and Carica papaya (Caricaceae) against the protozoan fish parasite Ichthyophthirius multifiliis.

The ciliate Ichthyophthirius multifiliis is among the most pathogenic parasites of fish maintained in captivity. In the present study, the effects of the crude methanolic extract of leaves of Mucuna pruriens and the petroleum-ether extract of seeds of Carica papaya against I. multifiliis were investigated under in vivo and in vitro conditions. Goldfish (Carassius auratus auratus) infected with the parasites were immersed for 72 h in baths with M. pruriens extract, and for 96 h in baths with C. papaya extract. There was a 90% reduction in numbers of I. multifiliis on fish after treatment in baths of each plant extract at 200 mg l(-1 )compared to untreated controls. Consequently, parasite-induced fish mortality was reduced significantly. A complete interruption of trophont recruitment was achieved by immersion in the M. pruriens extract. In vitro tests led to a 100% mortality of I. multifiliis in 150 mg/l M. pruriens extract, and in 200 mg/l of C. papaya extract after 6 h. Although the active constituents of the medicinal plant extracts are still unknown, we have demonstrated that they have potential for effective control of I. multifiliis.

Animals↗

Ant-colonized domatia on fruits of Mucuna interrupta (Leguminosae).

Domatia of myrmecophytes have been reported only in vegetative organs so far. In this paper, we report the first record of domatia formed on the legume surface of Mucuna interrupta. The domatium consists of 12-16 vertical lamellae which traverse the fruit surface obliquely to form long, narrow hollows, providing two ant species with a nest site.

Animals↗

Structural characterization of the N-glycans of gpMuc from Mucuna pruriens seeds.

Mucuna pruriens seeds are used in some countries as a human prophylactic oral anti-snake remedy. Aqueous extracts of M. pruriens seeds possess in vivo activity against cobra and viper venoms, and protect mice against Echis carinatus venom. It was recently demonstrated that the seed immunogen generating the antibody that cross-reacts with the venom proteins is a multiform glycoprotein (gpMuc), and the immunogenic properties of gpMuc seemed to mainly reside in its glycan chains. In the present study, gpMuc was found to contain only N-glycans. Part of the N-glycans could be released with peptide-(N (4)-(N-acetyl-beta -glucosaminyl)asparagine amidase F (PNGase F-sensitive N-glycans); the PNGase F-resistant N-glycans were PNGase A-sensitive. The oligosaccharides released were analyzed by a combination of MALDI-TOF mass spectrometry, HPLC profiling of 2-aminobenzamide-labelled derivatives and (1)H NMR spectroscopy. The PNGase F-sensitive N-glycans comprised a mixture of oligomannose-type structures ranging from Man(5)GlcNAc(2) to Man(9)GlcNAc(2), and two xylosylated structures, Xyl(1)Man(3)GlcNAc(2) and Xyl(1)Man(4)GlcNAc(2). The PNGase A-sensitive N-glycans, containing (alpha 1-3)-linked fucose, were identified as Fuc(1)Xyl(1)Man(2)GlcNAc(2) and Fuc(1)Xyl(1)Man(3)GlcNAc(2). In view of the determined N-glycan ensemble, the immunoreactivity of gpMuc was ascribed to the presence of core (beta 1-2)-linked xylose- and core alpha (1-3)-linked fucose-modified N-glycan chains.

Animals↗

Protection of Mucuna pruriens seeds against Echis carinatus venom is exerted through a multiform glycoprotein whose oligosaccharide chains are functional in this role.

In a previous paper we demonstrated that extracts of Mucuna pruriens seeds (MPE) protect mice against Echis carinatus venom (EV) by an immunological mechanism. In this paper we demonstrate that the MPE immunogen generating the antibody that cross-reacts with the venom proteins is a multiform glycoprotein (gpMuc) whose immunogenic properties mainly reside in its glycan-chains. The glycoprotein was purified from the protein extract of M. pruriens seeds using Concanavalin A affinity chromatography. Using 2-D gel electrophoresis it separated into seven isoforms having MWs in the range from 20.3 to 28.7 kDa and pIs from 4.8 to 6.5. N-terminal sequencing of these spots revealed close similarity since all of them contained the consensus sequence DDREPV-DT found in soybean Kunitz-type trypsin inhibitor. We suggest that gpMuc contains both N- and O-glycans. Mild alkaline treatment but not PNGase F led to loss of reactivity, indicating that O-glycans are probably involved in the antigenicity of gpMuc.

Amino Acid Sequence↗

The influence of kosmotropic and chaotropic salts on the functional properties of Mucuna pruriens protein isolate.

The influence of chaotropic and kosmotropic salts on Mucuna pruriens protein isolates was investigated. Protein solubility profile indicated that solubility was minimal at the isoelectric point of the protein isolate (4.0) while the solubility was maximal at pH 10.0 in all salt solutions. Chaotropes (I(-), ClO(4)(-) and SCN(-)) exhibit better protein solubility than the kosmotropes (SO(4)(2-), Cl(-) and Br(-)). Increase in protein solubility follows the Hofmeister series: NaSO(4)<NaCl<NaBr<NaI<NaClO(4)<NaSCN. Maximal water absorption capacity was recorded at low concentration of all the salts (0.1M). The water absorption capacity reduced as the concentration of the salts was increased to 2.0M. Kosmotropic salts exhibited better water absorption capacity than chaotropic salts. Both foaming capacity and stability were better in chaotropic salts compared with kosmotopic salts. Maximal foam capacities and stability were observed in protein solutions at 0.1M concentration. However the foam capacity and stability decreased as the concentration was increased further to 2.0M. The result indicates that emulsion activity index (EAI) and emulsion stability index (ESI) was reduced as the concentration of salts in each protein solution was increased. Protein solutions which contained NaSCN exhibited the highest emulsifying properties while those in Na(2)SO(4) recorded the lowest emulsifying properties. In addition, chaotropic salts had better emulsifying properties compared with kosmotropic salts. Increase in emulsifying activity follows the Hofmeister series. The least gelation concentration increased as the concentration of the salts increased. However chaotropic salts exhibited better gelation properties than kosmotropic salts.

Emulsions↗

Alkaloidal constituents of Mucuna pruriens seeds.

The seeds of Mucuna pruriens (L.) DC. after chemical analysis afforded four tetrahydroisoquinoline alkaloids which have been isolated for the first time from M. pruriens. Out of them, two are new whose structures have been elucidated by spectroscopic methods.

Alkaloids↗

Reduced allelopathic inhibition of lettuce (Lactuca sativa) growth caused by velvet bean (Mucuna pruriens) under 3D-clinorotation.

Allelopathy between Mucuna pruriens (velvet bean) and Lactuca sativa (lettuce) was studied under 3D-clinorotation. Growth of both roots and shoots of lettuce seedlings was suppressed by the presence of velvet bean. The degree of suppression was less on the clinostat compared to the normal static earth gravity. L-DOPA (L-3, 4-dihydroxyphenylalanine) is known to be a major substance in allelopathy of velvet bean. Amount of L-DOPA diffused out from a sintered filter paper into agar medium was compared between clinorotation and control group, and found no significant difference. It was concluded that some factors related to release, transport, and sensing phenomena of allelopathic substances may be responsible to the new findings in this study.

Biological Transport↗

Volatile allelopathy in velvet bean (Mucuna pruriens [correction of pruiens] L.) and gravity.

Allelopathy caused by volatile compounds was studied with velvet bean (Mucuna pruriens [correction of prueiens] L.). Preparatory experiment was conducted to identify the relationship between action of volatiles and gravity. The experimental protocol was defined and tested to study on volatile allelopathy of velvet bean. Root growth of lettuce (Lactuca sativa L.) seedlings was inhibited by volatile gas from velvet bean seedlings planted in Agripot. In this study, it was shown that ten-day velvet bean seedling could exhibit the volatile allelopathic ability. Growth chambers for volatile experiment were designed. Velvet bean seed can germinate in a 3-ml small vial.

Lactuca↗

Distribution of L-DOPA in the root of velvet bean plant (Mucuna pruriens L.) and gravity.

Velvet bean (Mucuna pruriens L.) has been found that the degree of suppression on the lettuce root growth by velvet bean was less on the 3D-clinorotation. The number and growth of adventitious root in velvet bean differed among the clinostated and control group. L-DOPA (L-3,4-dihydroxyphenylalanine) is known to be the major substance in the allelopathy of velvet bean plant, released from its root. Since L-DOPA is a precursor of melanin pigment, and is easily converted to melanin by oxidation, locality of L-DOPA production in the plant body can be seen through pigmentation. The amount of L-DOPA was analyzed by HPLC and LC-ESI/MS. The distribution of L-DOPA in the root was different among the ground control condition and pseudo-microgravity.

Gravitation↗

Effect of Mucuna urens (horse eye bean) on the gonads of male guinea-pigs.

The effect of Mucuna urens (seeds) on the gonads and sex accessory glands of male guinea-pigs was investigated. Sexually mature guinea-pigs of proven fertility were administered orally with 70 mg/kg and 140 mg/kg body weight of crude extract daily for 8 weeks respectively. Phytochemical screening of the seeds revealed the presence of alkaloids. No death or weight loss were observed during the duration of treatment. No pregnancy occurred in females mated with the treated males. Histological observations at high dose (140 mg/kg) showed complete degeneration of sperm in the testicular tubules. In some tubules, the acrosomal cap of the sperm cells was separated from the nuclei which underwent colour changes. In some tubules only the tails were left in the lumen. The spermatids, primary and secondary spermatocytes showed pycnosis while the morphology of spermatogonia and germinal epithelium appeared normal. Some epididymides were devoid of sperm while others contained degenerated spermatozoa and cell debris. In the prostate gland there was collapse of the villi and reduction of secretion in both the prostate and seminal vesicles. At low doses (70 mg/kg), there was spermatogenic arrest at spermatid stage. These observations have shown that M. urens is a potential male antifertility agent.

Animals↗

Blood chemistry of rats pretreated with Mucuna pruriens seed aqueous extract MP101UJ after Echis carinatus venom challenge.

The effect of a lethal Echis carinatus venom on serum enzyme levels and blood plasma coagulation parameters in rats pretreated with Mucuna pruriens seed aqueous extract MP101UJ (21 mg/kg body wt) 24 h and 3 wk before i.p venom injection (0.50 mg/kg rat) and rats injected with venom alone (0.50 mg/kg body wt) was investigated. The enzyme levels and coagulation parameter levels were measured 4 h after venom administration. The results showed that the increased enzymes lactate dehydrogenase (LDH), glutamic pyruvic transaminase (SGPT), creatinine kinase (CK) and changed coagulation parameters D-Dimer and Quick levels due to the venom effect were inhibited by M. pruriens seed aqueous extract MP101UJ in pretreated rats. Rats pretreated with a single dose (21 mg/kg and multiple doses 21 mg/kg rat) of extract MP101UJ maintained the normal enzyme levels and showed an anticoagulant effect as evidenced by the high PTT level which was also observed in venom treated animals. D-Dimer and Quick values were normal. However, the extract MP101UJ appeared to significantly inhibit the lethal venom induced myotoxic, cytotoxic and coagulation activities in experimental animals.

Animals↗

Effect of the alcohol extract of the seeds of Mucuna pruriens on free radicals and oxidative stress in albino rats.

In vitro and in vivo studies were made with an alcohol extract of the seeds of Mucuna pruriens (Fabaceae) to investigate its antioxidant property. In vitro studies were carried out in rat liver homogenate to investigate the chemical interaction of various phytochemicals with different species of free radicals. The effect was also checked on iron-induced lipid peroxidation, oxidation of GSH content, and its interaction with hydroxyl and superoxide radicals. There was no change on the rate of aerial oxidation of GSH content but it significantly inhibited FeSO(4) induced lipid peroxidation. It also inhibited the specific chemical reactions induced by superoxides and hydroxyl radicals. The removal of these species was through direct chemical interaction. An in vivo study on albino rats for 30 days showed no toxic effect up to a dose of 600 mg/kg body weight, on oral administration. There was no change in the level of TBA-reactive substances, reduced glutathione content and SOD activity in the liver. The activity of serum GOT, GPT and alkaline phosphatase was also unchanged. Thus it could be concluded that the alcohol extract of the seeds of M. pruriens has an antilipid peroxidation property, which is mediated through the removal of superoxides and hydroxyl radicals.

Alanine Transaminase↗

Certain functional properties of gums derived from some lesser known tropical legumes (Afzelia africana, Detarium microcarpum and Mucuna flagellipes).

Gum was extracted from some lesser known tropical legumes--Afzelia africana, Detarium microcarpum and Mucuna flagellipes. The extracts were evaluated for some functional properties. At a constant shear rate the apparent viscosity of the gums was directly proportional to the gum concentration. At 2% concentration, the gum dispersions showed an apparent viscosity of 41.24, 115.05 and 145.14 cps for A. africana, D. microcarpum and M. flagellipes gums respectively, measured at 174/sec and 25 +/- 1 degree C with the NV sensor of Haaké rotovisko viscometer. All the gums were found to contain D-galactose as a major monosaccharide. In addition, the presence of D-mannose, and D-glucose were indicated in both D. microcarpum and M. flagellipes while L-rhamnose, was indicated only in A. africana. A. africana showed significantly (p < 0.05) lower water absorption capacity and gelation properties than D. microcarpum and M. flagellipes. All the gums showed good emulsion properties. Emulsions of M. flagellipes gum showed little response to pH changes while A. africana and D. microcarpum gum showed better emulsion properties at acid pH than at alkaline pH.

Absorption↗

Mucuna pruriens: improvement of the biotechnological production of the anti-Parkinson drug L-dopa by plant cell selection.

Routinely grown cell suspension cultures of Mucuna pruriens L. (Fabaceae) were able to endogenously accumulate the anti-Parkinson drug L-dihydroxyphenylalanine (L-dopa) in the range between 0.2 and 2% on a dry weight (DW) basis. The green colour that developed in light-exposed cultures, appeared to be a suitable marker to select cells with an increased L-dopa biosynthesis and/or phenoloxidase activity. For this purpose, saccharose concentrations from 0 to 4% (w/v), and light intensities of 1,000 and 2,000 lux, were involved in the selection procedure. After 6 months, photomixotrophic callus cultures with a rapid growth and a high L-dopa content of 0.9% (DW) were obtained on 2% saccharose and under 1,000 lux. The cell suspensions, derived from these calli, accumulated up to 6% (DW) L-dopa, which was the highest stable content ever measured in cultures of M. pruriens. An L-dopa yield of approximately 1.2 g/l was calculated after 6 days of growth. In contrast, compared wtih the standard-grown parent cell line, the phenoloxidase activity, and consequently the bioconversion capacity as measured after entrapment in calcium alginate, of these high-producing cultures was approximately threefold lower.

Alginates↗

The biochemical composition and nutritional potential of the tribal pulse, Mucuna monosperma DC. ex Wight.

Two germplasms of the tribal pulse, Mucuna monosperma Dc. ex Wight, viz., Thirunelly Reserve Forest, Wynaad District (Kerala) and Siruvani Reserve Forest, Coimbatore District (Tamilnadu), were analysed for proximate composition, total (true) seed proteins, seed protein fractionation, amino acid composition, fatty acids, minerals and anti-nutritional factors. Crude proteins, crude lipids, ash and nitrogen free extractives constituted 23.10%, 4.36%, 3.80%, 61.74%, respectively, in Tamilnadu germplasm; whereas, in Kerala germplasm they constituted 20.13%, 8.99%, 3.60% and 61.69%, respectively. The calorific values of 100 g dry matter of seed material are 408.19 KCal (Kerala) and 378.60 KCal (Tamilnadu) germplasms. Essential amino acids like methionine, cystine, threonine and valine were found to be limiting in the seed proteins of both the germplasms. The fatty acids like lauric acid and palmitic acid are found to be common in both the germplasm seed samples, while linoleic and oleic acids in Kerala germplasm and stearic acid in Tamilnadu germplasm are present. The Tamilnadu germplasm seems to be a rich source of sodium, magnesium, zinc, manganese, copper and iron; whereas the Kerala germplasm seems to be a rich source of phosphorus. Antinutritional substances like total free phenols, tannins, L-DOPA, trypsin inhibitor activity and phytohaemagglutinating activity also were investigated.

Amino Acids↗

The biochemical composition and nutritional potential of the tribal pulse, Mucuna gigantea (Willd) DC.

Seeds of the tribal pulse, Mucuna gigantea, were analysed for proximate composition, total (true) seed proteins, seed protein fractionation, amino acid composition, minerals and antinutritional factors. Crude proteins, crude lipids, ash and nitrogen free extractives constituted 30.62%, 9.03%, 5.99% and 42.79%, respectively. The calorific values of 100 g dry matter of seed material is 374.91 kCal. The essential amino acids, leucine + isoleucine, are present in relatively large quantities. The seeds are rich in minerals like K, Ca, Mg and Fe. Antinutritional substances like total free phenols and tannins, trypsin inhibitor activity and haemagglutinating activity also were investigated.

Albumins↗

Potentials of gum from Detarium microcarpum (DM) and Mucuna flagellipes (MF) seeds as raw beef burger stabilizers.

Raw beef burgers containing graded levels (0.25, 0.5, 0.75. and 1.0%) of polysaccharide gums extracted from Detarium microcarpum (DM) and Mucuna flagellipes (MF) were produced. Unstabilized beef burgers and beef burgers containing gum tragacanth (TR) were also produced simultaneously to serve as control. The raw beef burgers were evaluated for selected physicochemical and sensory properties so as to assess the stabilization potentials of DM and MF polysaccharide gums relative to the conventional TR. Beef burgers containing the polysaccharide gums had significantly (P < 0.05) lower shrinkage, higher water holding capacity (WHC), and better stability under ambient conditions (27 +/- 1 degrees C and RH 90.6%) then the unstabilized burgers. Among the stabilized burgers, DM gum burgers had lower (P < or = 0.05) shrinkage and higher WHC than MF and TR gums burgers. All the gum-stabilized beef burgers required 9-26% higher compression force indicating a less friable product than the unstabilized burgers. At 0.25 and 0.5% levels of gum incorporation DM gum burgers had comparable compression force with TR gum burgers and this was lower (P < or = 0.05) than the compression force of MF gum burgers. Sensory panel results indicated a lower score for finger feel in gum-stablized burgers than the unstabilized burgers. DM and TR gum burgers had comparable scores with TR gum burgers in all the sensory attributes tested. Overall acceptability score showed that all the gum-stabilized beef burgers were acceptable. It was evident from the study that polysaccharide gums from DM and MF seeds can serve as effective stabilizers in beef burgers without adverse effect on the quality of the product.

Animals↗

Cyclodextrin-facilitated bioconversion of 17 beta-estradiol by a phenoloxidase from Mucuna pruriens cell cultures.

After complexation with beta-cyclodextrin, the phenolic steroid 17 beta-estradiol could be ortho-hydroxylated into a catechol, mainly 4-hydroxyestradiol, by a phenoloxidase from in vitro grown cells of Mucuna pruriens. By complexation with beta-cyclodextrin the solubility of the steroid increased from almost insoluble to 660 microM. The bioconversion efficiency after 72 hr increased in the following order: freely suspended cells (0%), immobilized cells (1%), cell homogenate (6%), phenoloxidase preparation (40%). Mushroom tyrosinase converted 17 beta-estradiol, as a complex with beta-cyclodextrin, solely into 2-hydroxyestradiol, with a maximal yield of 30% after 6-8 hr. Uncomplexed 17 beta-estradiol was not converted at all in any of these systems.

Biotransformation↗