Research Update:Alfalfa Medicago sativa.

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   Phytochemical info of Alfalfa Extract:

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 Definition:Alfalfa Extract are majorly composed of
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  Research Update:Alfalfa Medicago sativa

  Crystal Structure of Vestitone Reductase from Alfalfa (Medicago sativa L.).:J Mol Biol. 2007 May 25;369(1):265-76. Epub 2007 Mar 21.Shao H, Dixon RA, Wang X.Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA.

 Isoflavonoids are commonly found in leguminous plants, where they play important roles in plant defense and have significant health benefits for animals and humans. Vestitone reductase catalyzes a stereospecific NADPH-dependent reduction of (3R)-vestitone in the biosynthesis of the antimicrobial isoflavonoid phytoalexin medicarpin. The crystal structure of alfalfa (Medicago sativa L.) vestitone reductase has been determined at 1.4 A resolution. The structure contains a classic Rossmann fold domain in the N terminus and a small C-terminal domain. Sequence and structural analysis showed that vestitone reductase is a member of the short-chain dehydrogenase/reductase (SDR) superfamily despite the low levels of sequence identity, and the prominent structural differences from other SDR enzymes with known structures. The putative binding sites for the co-factor NADPH and the substrate (3R)-vestitone were defined and located in a large cleft formed between the N and C-terminal domains of enzyme. Potential key residues for enzyme activity were also identified, including the catalytic triad Ser129-Tyr164-Lys168. A molecular docking study showed that (3R)-vestitone, but not the (3S) isomer, forms favored interactions with the co-factor and catalytic triad, thus providing an explanation for the enzyme's strict substrate stereo-specificity.

  Qualitative and quantitative polymerase chain reaction assays for an alfalfa (Medicago sativa)-specific reference gene to use in monitoring transgenic cultivars.:J Agric Food Chem. 2007 Apr 18;55(8):2918-22. Epub 2007 Mar 20.Alexander TW, Reuter T, McAllister TA.Agriculture and Agri-Food Canada Research Centre, P.O. Box 3000, Lethbridge, Alberta, Canada.

 Genetically modified (GM) alfalfa (Medicago sativa) was marketed for the first time in 2005. For countries with established thresholds for GM plants, methods to detect and quantify their adventitious presence are required. We selected acetyl CoA carboxylase as a reference gene for the detection and quantification of GM alfalfa. Two qualitative polymerase chain reaction (PCR) assays (Acc1 and Acc2) were designed to detect alfalfa. Both were specific to alfalfa, amplifying DNA from 12 separate cultivars and showing negative results for PCR of 15 nonalfalfa plants. The limits of detection for Acc1 and Acc2 were 0.2 and 0.01%, respectively. A quantitative real-time PCR assay was also designed, having high linearity (r > 0.99) over alfalfa standard concentrations ranging from 100 to 2.0 x 10(5) pg of alfalfa DNA per PCR. The real-time PCR assay was effective in quantifying alfalfa DNA from forage- and concentrate-based mixed diets containing different amounts of alfalfa meal.

  Flavonoids from barrel medic (Medicago truncatula) aerial parts.:J Agric Food Chem. 2007 Apr 4;55(7):2645-52. Epub 2007 Mar 10.Kowalska I, Stochmal A, Kapusta I, Janda B, Pizza C, Piacente S, Oleszek W.Department of Biochemistry, Institute of Soil Science and Plant Cultivation, ul. Czartoryskich 8, 24-100 Pulawy, Poland.

 Twenty-three flavonoids have been identified in the aerial parts of barrel medic, and their structures were established by spectrometric and spectroscopic (ESI-MS/MS and NMR) techniques. Eight of the identified compounds, including apigenin 7-O-beta-D-glucuronopyranosyl-(1-->3)-O-beta-D-glucuronopyranosyl-(1-->2)-O-beta-D-glucuronopyranoside, apigenin 7-O-[2'-O-sinapoyl-beta-D-glucuronopyranosyl-(1-->2)-O-beta-D-glucuronopyranoside], apigenin 7-O-{2-O-feruloyl-[beta-D-glucuronopyranosyl-(1-->3)]-O-beta-D-glucuronopyranosyl-(1-->2)-O-beta-D-glucopyranoside}, chrysoeriol 7-O-[beta-D-glucuronopyranosyl-(1-->2)-O-beta-D-glucuronopyranoside, chrysoeriol 7-O-{2'-O-p-coumaroyl-[beta-D-glucuronopyranosyl-(1-->3)]-O-beta-D-glucuronopyranosyl(1-->2)-O-beta-D-glucuronopyranoside}, tricin 7-O-beta-D-glucuronopyranosyl-4'-O-glucopyranoside, tricin 7-O-[2'-O-feruloyl-beta-D-glucuronopyranosyl-(1-->2)-O-beta-D-glucopyranoside], and tricin 7-O-{2'-O-p-coumaroyl-[beta-D-glucuronopyranosyl-(1-->3)]-O-beta-D-glucuronopyranosyl(1-->2)-O-beta-D-glucuronopyranoside}, have not been reported before in the plant kingdom. Additionally, the presence of two luteolin, three apigenin, one chrysoeriol, and six tricin glycosides, previously identified in alfalfa (Medicago sativa), was confirmed in M. truncatula. Moreover, besides the above flavones, the aerial parts of this species contained three flavonols including rutin, laricitrin 3,7,5'-triglucoside, and laricitrin 3,5'-diglucoside.


  Evaluation of the estrogenic effects of legume extracts containing phytoestrogens.:J Agric Food Chem. 2003 Apr 9;51(8):2193-9.

 Seven legume extracts containing phytoestrogens were analyzed for estrogen 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.

  Treatment of neurovegetative menopausal symptoms with a phytotherapeutic agent.:Minerva Ginecol. 1998 May;50(5):207-11. Istituto di Ginecologia e Ostetricia, Universita degli Studi-Siena.

 Menopausal symptoms include hot flushes, insomnia, nocturnal sweating, dizziness, headaches and palpitations. These symptoms reflect adaptation of the body to estrogen deprivation which affects various central neurotransmitters. METHODS: In this study, the efficacy has been tested of a plant product based on extracts of the leaves of Salvia officinalis (sage) and Medicago sativa (alfalfa) in the treatment of hot flushes in 30 menopausal women with these symptoms. RESULTS: Hot flushes and night sweating completely disappeared in 20 women: four women showed good improvement and the other six showed a reduction in symptoms. GnRH and TRH tests were performed in 8 women to evaluate TSH and Prl responses before and after 3 months of therapy. The plants product induced a significant increase in Prl and TSH response to TRH. Basal levels of estradiol, LH, FSH, Prl and TSH were unchanged. CONCLUSIONS: Alfalfa and sage seem to have a central slight antidopaminergic action without side effects and appear to be an effective combination in the treatment of menopausal symptoms.

  Soy and alfalfa phytoestrogen extracts become potent low-density lipoprotein antioxidants in the presence of acerola cherry extract.:J Agric Food Chem. 2001 Jan;49(1):308-14.

 Postmenopausal women have an increased risk of coronary heart disease. Oxidation of low-density lipoprotein (LDL) has been implicated in atherogenesis, and the presence of modified LDL (LDL(-)) in plasma appears to represent LDL oxidation in vivo. Because previous studies have demonstrated a strong antiatherogenic effect of estrogen due to its antioxidant activity and similar antioxidant activity was found for specific isoflavones derived from soy extract, the antioxidant activity of a phytoestrogen extract derived from soy and alfalfa was studied. Copper-mediated LDL oxidation was inhibited in the presence of soy and alfalfa extracts, and this effect was further enhanced in the presence of acerola cherry extract, which is rich in ascorbic acid. Male rabbit aortic endothelial cells pretreated with soy extract were resistant to the toxic effects of high levels of LDL and LDL(-), and a lesser, but significant protection, was also afforded by alfalfa extract. Cell-mediated oxidation of LDL, measured by LDL(-) formation, was inhibited in the presence of soy extract but not alfalfa extract. However, in the presence of acerola cherry extract, both soy and alfalfa extracts potently inhibited the formation of LDL(-). These findings show that acerola cherry extract can enhance the antioxidant activity of soy and alfalfa extracts in a variety of LDL oxidation systems. The protective effect of these extracts is attributed to the presence of flavonoids in soy and alfalfa extracts and ascorbic acid in acerola cherry extract, which may act synergistically as antioxidants. It is postulated that this synergistic interaction among phytoestrogens, flavonoids, and ascorbic acid is due to the "peroxidolitic" action of ascorbic acid, which facilitates the copper-dependent decomposition of LDL peroxides to nonradical products; this synergy is complemented by a mechanism in which phytoestrogens stabilize the LDL structure and suppress the propagation of radical chain reactions. The combination of these extracts markedly lowers the concentrations of phytoestrogens required to achieve significant antioxidant activity toward LDL.


  Alfalfa and Kelp Combination:natural dietic,benefit metabolism and stimulates protein,increased muscle for bodybuilders

 Alfalfa and Kelp are an extremely valuable source of natural nutrients such as minerals and natural iodine which is known to be a natural dietic.When taken in combination there is a beneficial effect upon the body's metabolism which stimulates a more effective use of the protein consumed.This metabolic effect is especially useful to bodybuilders and athletes training for increased muscle size and added strength.
 

  Antimicrobial activity of saponins from Medicago sp.: structure-activity relationship.:

 The antimicrobial activity of saponins from Medicago sativa, M. arborea and M. arabica against a selection of medically important yeasts, Gram-positive and -negative bacteria was investigated. Structure-activity growth inhibitory effects of related prosapogenins and sapogenins are also described. Increasing antibiotic activity was observed going from the saponin extracts to the sapogenin samples, suggesting that the sugar moiety is not important for the antimicrobial efficacy. Activity was especially high against Gram-positive bacteria (Bacillus cereus, B. subtilis, Staphylococcus aureus and Enterococcus faecalis) with M. arabica being the species showing a broader spectrum of action. Discrete antifungal activity was also observed, mainly against Saccharomyces cerevisiae. The observed antimicrobial properties of M. sativa and M. arborea were related to the content of medicagenic acid, while hederagenin seems to contribute to the bioactivity of M. arabica total sapogenins.

  Cholesterol levels and sensory characteristics of meat from broilers consuming moderate to high levels of alfalfa.:

 There is an urgent need in industrialized societies to develop novel products that can lower human dietary cholesterol intake. Dehydrated alfalfa is a good source of hypocholesterolemic compounds such as saponins. Whether consumption of alfalfa by chickens would decrease the cholesterol content of broiler meat remains, however, to be established. We determined meat quality characteristics, lipid and cholesterol contents, and consumer preference of broiler meat derived from production systems based on restricted feeding of a commercial diet combined with provision of free-choice dehydrated alfalfa. Results demonstrated that it was possible to produce chicken breast meat with reduced cholesterol content. In addition, total lipids in chicken meat were significantly decreased when a higher level of restriction was applied. Members of a 50-person consumer panel preferred meat from animals not consuming or consuming moderate levels of alfalfa. Those members that preferred meat from animals consuming the higher percentage of alfalfa identified taste as the primary attribute influencing that decision. Together the results suggest that it is possible to develop novel broiler production systems that will produce leaner meat that is acceptable to consumers and has a reduced cholesterol content.

  The effect of alfalfa-corn diets on cholesterol metabolism and gallstones in prairie dogs.:

 Cholesterol gallstones were present in prairie dogs fed alfalfa plus corn with and without exogenous cholesterol (0.4%). The diets fed to the animals for eight weeks contained alfalfa plus corn in fixed proportions of 50:50, 85:15 and 15:85 (w/w). At sacrifice, all animals were healthy but had not gained weight; no deaths occurred during the experiment. Cholesterol gallstones were present in all groups. In the absence of exogenous cholesterol, the highest stone incidence was found in the animals which received the lowest fiber (highest corn) diets (alfalfa plus corn, 50:50, 67%; alfalfa plus corn, 15:85, 83%). Cholesterol gallstone incidence was 100% when exogenous cholesterol was added to the alfalfa plus corn diets (50:50 and 15:85). No pigment gallstones were detected in any animal. Liver and plasma cholesterol concentrations were highest in the animals receiving alfalfa plus corn (15:85) plus 0.4% cholesterol (4.29 mg/g, and 356 mg/dl, respectively). These values were lowest in animals receiving 85% alfalfa plus 15% corn without cholesterol (2.19 mg/g and 88 mg/dl, respectively). Lithogenic indices were below 1.00 in all groups. Biliary bile acids were mainly amidates of cholic acid and chenodeoxycholic acid, with the former predominating. Thus, gallstones can be formed in prairie dogs in the absence of exogenous cholesterol; gallstone incidence is reduced by dietary fiber.

  Antiatherosclerotic effects of alfalfa meal ingestion in chicks: a biochemical evaluation.:

 Feeding Alfalfa seed extract to chicks resulted in significant reduction of total cholesterol, phospholipid, triglyceride, LDL-Cholesterol and VLDL-cholesterol. Simultaneously, an increased in the HDL-cholesterol/total cholesterol ratio occurred with reduction in total cholesterol and phospholipid contents of liver and (ventricular) muscle of the heart. Results obtained with alfalfa seed extract ingestion were evaluated with a standard drug, compound, clofibrate.


  Cholesterol and bile acid balance in Macaca fascicularis. Effects of alfalfa saponins.:

 We determine the effects of alfalfa top saponins on cholesterol and bile acid balance in eight cynomolgus macaques (Macaca fascicularis). The monkeys ate semipurified food containing cholesterol with or without added saponins. The saponins decreased cholesterolemia without changing the levels of high density lipoprotein-cholesterol; hence, they reduced the total cholesterol/high density lipoprotein-cholesterol ratio. Furthermore, they decreased intestinal absorption of cholesterol, increased fecal excretion of endogenous and exogenous neutral steroids and bile acids, and decreased the percent distribution of fecal deoxycholic and lithocholic acids. The fecal excretion of fat was also slightly increased, but steatorrhea did not occur. We saw no signs of toxicity in the monkeys after 6 or 8 wk of saponin ingestion. The data suggest that alfalfa top saponins may be of use in the treatment of patients with hypercholesterolemia, but long-term studies on possible toxicity are needed before this therapy can be recommended for humans.

  Comparative effects of alfalfa saponins and alfalfa fiber on cholesterol absorption in rats.:

 Intestinal absorption of cholesterol was measured in control rats fed semipurified diets and in rats fed alfalfa meal, in which saponins had been previously extracted, or this extracted material plus alfalfa saponins. A dose of 2 mg radioactive cholesterol was administered intragastrically, and fecal excretion of labeled neutral steroids measured. Absorption of cholesterol was about 76% in control animals, and about 47% in alfalfa-red rats. Extraction of saponins from alfalfa eliminated the cholesterol absorption-lowering effect, while addition of 0.26% alfalfa saponins to the extracted alfalfa restored its activity. The results demonstrate that alfalfa saponins are responsible for the effect of alfalfa meal in reducing cholesterol absorption, and that alfalfa fiber is not involved in this activity.

  Immunopotentiating effects of polysaccharides isolated from Medicago sativa L:

 In vitro experiments showed that 250-500 micrograms.ml-1 of the polysaccharides isolated from Medicago sativa L (MSP) increased mouse lymphocyte uptake of [3H]thymidine (TdR) induced by PHA-p (0.15%), Con A (5 micrograms.ml-1), LPS (20 micrograms.ml-1), and PWM (2 micrograms.ml-1) at maximum by 5.13 +/- 0.81, 0.75 +/- 0.08, 4.52 +/- 0.36, and 0.86 +/- 0.24 folds, respectively. After MSP ip for 5 d, the average count of mouse splenic lymphocytes in 125 mg.kg-1 x d-1 group increased from 1.4 +/- 0.4 (x 10(7))/mouse to 3.6 +/- 0.5 (x 10(7))/mouse; in 250 mg.kg-1 x d-1 dose group increased to 13.5 +/- 0.3 (x 10(7)). MSP 125 mg.kg-1 x d-1 or 250 mg.kg-1 x d-1 ip for 5 d partially or completely antagonized the effect of cyclophosphamide (200 mg.kg-1 x d-1 x 1 d) on lymphocyte decrease, the average number of lymphocytes/mouse recovered from 1.04 +/- 0.23 (x 10(7)) to 2.1 +/- 0.4 (x 10(7)) and 2.9 +/- 0.6 (x 10(7)), respectively. In addition, the lymphocyte reactivity to Con A in MSP ip 125 mg.kg-1 x d-1 x 5 d group enhanced by 60%, in 250 mg.kg-1 x d-1 x 5 d group by 156%. The abilities of PWM-induced antibody production were elevated 51% and 78%, respectively.

  Effects of L-canavanine on T cells may explain the induction of systemic lupus erythematosus by alfalfa.:

 Alfalfa sprouts can induce systemic lupus erythematosus (SLE) in monkeys. This property of alfalfa sprouts has been attributed to their non-protein amino acid constituent, L-canavanine. Occurrence of autoimmune hemolytic anemia and exacerbation of SLE have been linked to ingestion of alfalfa tablets containing L-canavanine. In this report we show that L-canavanine has dose-related effects in vitro on human immunoregulatory cells, which could explain its lupus-inducing potential. These effects include: 1) diminution of the mitogenic response to both phytohemagglutinin and concanavalin A but not to pokeweed mitogen, as determined in both thymidine incorporation and cell cycle studies, and 2) abrogation of concanavalin A-induced suppressor cell function, which results in increased release of both IgG and DNA binding activity into supernatants by cells from normal subjects and SLE patients. These immunoregulatory effects of L-canavanine may explain the induction or exacerbation of SLE by alfalfa.

  The 'pterocarpan synthase' of alfalfa: association and co-induction of vestitone reductase and 7,2'-dihydroxy-4'-methoxy-isoflavanol (DMI) dehydratase, the two final enzymes in medicarpin biosynthesis.:

 Vestitone reductase and 7,2'-dihydroxy-4'-methoxy-isoflavanol (DMI) dehydratase are the two final enzymes in medicarpin biosynthesis in alfalfa (Medicago sativa). Although two independent enzymes, vestitone reductase and DMI dehydratase can be loosely associated in low ionic strength buffers, presumably by a weak protein-protein interaction. The activities of vestitone reductase and DMI dehydratase increased approximately 3-fold 6 hours after elicitor treatment in alfalfa suspension cell culture. The activities remained at maximal levels for 40 hours, correlating with a steady increase in the medicarpin content of the cells. Medicarpin produced in vitro from vestitone by the action of vestitone reductase and DMI dehydratase was found to be (-)-medicarpin (6aR,11aR-medicarpin), possessing the same stereochemistry as medicarpin produced in vivo.


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  Scientific References:

  1.Research Update:Alfalfa Medicago sativa.
  2.Alfalfa Medicago sativa.Alfalfa Extract.