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Role of nitric oxide in the vasorelaxant and hypotensive effects of extracts and purified tannins from Geum japonicum.

Crude extracts and three purified tannins from Geum japonicum Thunberg (Rosaceae) were examined for relaxant effects in isolated rat thoracic aorta and for hypotensive effects in anesthetized normotensive and hypertensive rats. The acetone extract and the butyl alcohol extract of Geum japonicum at a cumulative concentration of 30mug/ml potently relaxed phenylephrine-precontracted aortic rings by 73+/-5% and 80+/-7%, respectively, without affecting the resting tension of these vessels. Removal of the vascular endothelium, inhibition of nitric oxide (NO) synthase with N(omega)-nitro-l-arginine (l-NA) or inhibition of cGMP biosynthesis with methylene blue all abolished the vasorelaxant effects of the Geum japonicum extracts. Addition of l-arginine, the substrate for NO biosynthesis, reversed the inhibitory effects of l-NA. Similar vasorelaxant effects of 82+/-10%, 61+/-8% and 82+/-14%, were observed with the purified tannins, penta-O-galloyl-beta-glucoside, casuariin and 5-desgalloylstachyurin, respectively, at a cumulative concentration of 10muM. Intravenous injection of the butyl alcohol extract of Geum japonicum at a cumulative dose of 2.5mg/kg into both hypertensive and normotensive rats resulted in a marked reduction in the mean arterial blood pressure by 46+/-6% and 34+/-7%, respectively, which was abolished by prior injection of l-NA. Therefore, these results suggest that tannins may be responsible for the vasorelaxant and hypotensive effects of Geum japonicum, mediated via endogenous NO and subsequent cGMP formation. The data suggest that extracts of Geum japonicum may have potential use as new anti-hypertensive agents for lowering arterial blood pressure in hypertensive patients.

Animals↗

Antimutagenicity of hydrolyzable tannins from Terminalia chebula in Salmonella typhimurium.

A tannin fraction (TC-E) from the dried fruit pulp of Terminalia chebula was obtained by successfully extracting with 95% ethyl alcohol and ethyl acetate. TC-E was subjected to silica gel chromatography which yielded four fractions, viz., TC-EI, TC-EII, TC-EIII and TC-EIV. Thin layer chromatography (TLC) and 13C-NMR revealed that TC-EI was gallic acid (GA) derivative while the other fractions were tannin in nature. TC-E and its fractions were evaluated for their antimutagenic potential against two direct-acting mutagens, 4-nitro-o-phenylenediamine (NPD) and 4-nitroquinoline-N-oxide (4NQNO), and S9-dependent mutagen, 2-aminofluorene (2AF) in TA98 and TA100 strains of Salmonella typhimurium. The study revealed that the extract (TC-E) and its fractions were highly significant against S9-dependent mutagen, 2AF. The effect was found to be more or less corresponding with the nature of the fractions, as the monomeric TC-EI (a GA derivative) was least effective as compared to other fractions which were oligomeric, and the order of their effectiveness as per their IbD50 value being TC-EIV (8.9 micrograms)>TC-EIII (17.8 micrograms)>TC-EII (45 micrograms)>TC-EI (320 micrograms) in TA98; TC-EIV being 40 times more effective than TC-EI in inhibiting his+ revertants. A similar effect was noticed in TA100 too, where TC-EI was the least effective and TC-EII had the maximum effect. A similar result was noticed when the antimutagenicity of GA (a monomeric) was compared with tannic acid (TA, an oligomeric). However, chebula tannins were found to be partly effective against NPD but not at all effective against 4NQNO.

Antimutagenic Agents↗

Effect of bioactive compounds from Sainfoin ( Onobrychis viciifolia Scop.) on the in vitro larval migration of Haemonchus contortus: role of tannins and flavonol glycosides.

Anthelmintic bioactivity against gastrointestinal nematodes has been associated with leguminous forages supporting the hypothesis of a role of condensed tannins. However, the possibility that other compounds might also been involved has received less consideration. Using bio-guided fractionation, the current study aimed at characterizing the biochemical nature of the active compounds present in sainfoin (Onobrychis viciifolia ), previously identified as an anthelmintic leguminous forage. The effects of sainfoin extracts were evaluated on 3rd-stage larvae (L3) of Haemonchus contortus by using a larval migration inhibition (LMI) assay. Comparison of extracts obtained with several solvent systems showed that the bioactivity was associated with the 70ratio30 acetone/water extract. Further fractionation of the later allowed the separation of phenolic compounds. By use of a dialysis method, compounds were separated with a molecular weight cut-off of 2000 Da. The in vitro anthelmintic effect of the fraction with condensed tannins was confirmed. In the fraction containing molecules of MW <2000 Da, 3 flavonol glycosides were identified as rutin, nicotiflorin and narcissin. At 1200 mug/ml, each inhibited significantly the migration of larvae. Addition of polyvinyl pyrrolidone (PVPP) to both fractions before incubation restored larval migration. These results confirmed the role of both tannins and flavonol glycosides in the anthelmintic properties of sainfoin.

Animals↗

Collagen fiber immobilized Myrica rubra tannin and its adsorption to UO2(2+).

Tannins, which are rich in ortho-hydroxyl groups, have a high affinity for UO2(2+). In this paper, Myrica rubra tannin was immobilized on collagen fiber by an aldehydic cross-linking reaction to prepare a novel adsorbent for uranium (UO2(2+)) recovery from wastewater. The adsorption equilibrium, the adsorption kinetics, and the effects of temperature and pH on the adsorption equilibrium were investigated in detail. It was found that the Myrica rubra tannin immobilized on collagen fiber exhibits an excellent adsorption capacity for UO2(2+). The adsorption capacity at 293 K and pH 5.0 was as high as 1.19 mmol UO2(2+)/g (283.3 mgU/g) when the initial concentration of UO2(2+) in solution was 7.5 mmol/L. The adsorption isotherms could be described by the Freundlich equation, and the increase of temperature promoted the adsorption to UO2(2+) . The adsorption kinetics data were fitted very well by the pseudosecond-order rate model, and the equilibrium adsorption capacity calculated by the pseudo-second-order rate model was almost the same as that determined by the actual measurement with the error < or = 4%. The pH has a significant effect on the adsorption process. According to our experiments, the suitable pH scope should be 5-8.

Adsorption↗

Effects of wine phenolics and sorghum tannins on tyrosinase activity and growth of melanoma cells.

In this study, three different phenolic (anthocyanin, other flavonoid, and phenolic acid) fractions from wine and a condensed tannin preparation from sorghum were tested for their effects on melanogenesis of normal cells and growth of human melanoma cells. The wine phenolic fractions decreased melanogenic activity (tyrosinase activity) at concentrations that resulted in a slight variation in melanocyte viability. Sorghum tannins, however, increased melanogenic activity, although no increase was found in total melanin at the concentrations that least affect melanocyte viability. Incubation of human melanoma cells with the wine fractions and sorghum tannins resulted in a decrease in colony formation, although the effect was not dose dependent in all cases. These results suggest that all of these phenolic fractions have potential as therapeutic agents in the treatments of human melanoma, although the mechanisms by which cellular toxicity is effected seem to be different among the fractions.

Anthocyanins↗

Determination of poly(ethylene glycol)-binding to browse foliage, as an assay of tannin, by near-infrared reflectance spectroscopy.

Nutritionists are interested in functional assays of tannins that do not require time-consuming and expensive extraction, such as the (14)C-labeled poly(ethylene glycol) (PEG)-binding (PEG-b) assay. This paper reports the application of near-infrared (NIR) spectroscopy to determine the percentage of PEG binding, in place of the (14)C-labeled PEG-b assay of tannin, in Mediterranean woodland vegetation. Calibration was done with 53 samples from 14 species and was validated on 25 samples from 10 species. PEG-b ranged between 1.4 and 20.7% in the samples. The calibration obtained by using the modified partial least-squares (MPLS) method, with all wavelengths in the 1100-2500 nm range combined, and the validation were reasonably linear (R (2) = 0.96 and 0.91, respectively). The accuracies, estimated from the standard errors of cross-validation and prediction, were +/-1.6 and +/-1.7% PEG-b, respectively. The NIRS-aided procedure proposed here can serve as an accurate, inexpensive, time-saving, and environment-friendly functional assay of tannin in Mediterranean browse.

Animal Nutritional Physiological Phenomena↗

Iron and zinc in vitro availability in pearl millet flours (Pennisetum glaucum) with varying phytate, tannin, and fiber contents.

Simulations of gastro-intestinal digestion, used to estimate in vitro iron and zinc availability, were performed on two kinds of samples: (i) samples with decreased phytate contents from whole pearl millet flour and (ii) nondephytinized or dephytinized samples from two pearl millet grain fractions, a decorticated fraction with low fiber and tannin contents and a bran fraction with high fiber and tannin contents. Iron and zinc in vitro availabilities of whole pearl millet flour were significantly improved by phytate degradation, even if the IP6 were not all degraded. Total dephytinization of decorticated fraction led to a marked increase in iron and zinc in vitro availabilities, but that of bran fraction had no effect on either iron or zinc in vitro availability. Even if phytates are involved in reducing in vitro iron and zinc availability in pearl millet flour, fibers and tannins play an important role by chelating a high proportion of iron and zinc in grain hulls.

6-Phytase↗

Determination of tannin in green tea infusion by flow-injection analysis based on quenching the fluorescence of 3-aminophthalate.

A flow-injection analytical system was developed to determine tannin content in green tea infusions. The flow-injection system is based on measuring the quenching effect of tannin on the fluorescence of 3-aminophthalate. Fluorophore was obtained by auto-oxidation of luminol during solution preparation. System performance was satisfactory for routine analysis (sample throughput >20 h(-1); linear dynamic range for tannic acid, 0.005-0.3 mg/mL; linear dynamic range for green tea tannin, 0.02-1.0 mg/mL; CV < 3%). The flow-injection method is immune from interference by coexisting ascorbate in green tea infusion. Analytical results were verified by the ferrous tartrate method, the Japanese official analytical method.

Ascorbic Acid↗

Accumulation and extractability of grape skin tannins and anthocyanins at different advanced physiological stages.

Quantitative and qualitative modifications of tannins and anthocyanins in grape skin were investigated at different dates of harvest, from berries sorted on the basis of their density. Free anthocyanins accumulated until 170 g/L of sugars in pulp before undergoing a slight decrease. Changes in anthocyanin composition were observed with increasing sugar levels in the pulp that reflected structural differences between classes of anthocyanins. The proportion of methoxylated anthocyanins continued to increase in the skin as sugar accumulated while the proportion of coumaroylated anthocyanins initially increased (up to 200 g/L of sugars in the pulp) and then rapidly decreased. In comparison, no major quantitative nor qualitative change was observed for tannins, except for a slight increase of the mean degree of polymerization. Whatever the physiological stage of the pulp, the extraction yield of skin phenolics into hydroalcoholic solution for 5 h was lower than 77% for anthocyanins and 38% for proanthocyanidins. For both classes of compounds, no clear evolution in these extraction yields could be observed as sugars accumulated in pulp (from 162.6 to 275.0 g/L). Nevertheless, some structural features within each family of compounds significantly influenced extractability, for example, a lower extraction yield for coumaroylated anthocyanins and for tannins with a high degree of polymerization. Finally, no direct relationship could be found in extraction media between the amounts of all red pigments (measured in acidic conditions) and the color intensity at 520 nm (measured in wine-like model solutions).

Anthocyanins↗

Optimization of tannin extraction from infant foods.

The design of experiments (DOE) was used in the development of a laboratory procedure for the extraction of tannins from three infant food types comprising different ingredients of vegetable origin and meat. The diversity of vegetables included in the product formulas required the use of DOE to establish parameters that maximize the recovery of tannins using a central composite rotatable design. Once the experimental results from the DOE were obtained, response surface methodology was used to find the best analytical conditions for samples comprising different ingredients. Sample weight was found to be a critical factor in tannin extraction from foods. Different optimal conditions were obtained for samples including soya in the formula.

Infant Food↗

Anti-AIDS agents, 2: Inhibitory effects of tannins on HIV reverse transcriptase and HIV replication in H9 lymphocyte cells.

Nine tannins, including gallo- and ellagitannins, were evaluated as potential inhibitors of HIV replication. 1,3,4-Tri-O-galloylquinic acid [1], 3,5-di-O-galloyl-shikimic acid [2], 3,4,5-tri-O-galloylshikimic acid [3], punicalin [6], and punicalagin [7] inhibited HIV replication in infected H9 lymphocytes with little cytotoxicity. Two compounds, punicalin and punicacortein C [8], inhibited purified HIV reverse transcriptase with ID50 of 8 and 5 microM, respectively. Further studies with H9 lymphocytes indicated that chebulagic acid [5] and punicalin did not inactivate virus directly. However, 1,3,4-tri-O-galloylquinic acid and 3,5-di-O-galloylshikimic acid were more effective inhibitors under those conditions. All tannins appear to inhibit virus-cell interactions. Thus, inspite of their anti-RT activity, the mechanism by which tannins inhibit HIV may not be associated with this enzyme.

Antiviral Agents↗

Antitumor agents, 129. Tannins and related compounds as selective cytotoxic agents.

Fifty-seven tannins and related compounds, including gallotannins, ellagitannins, and condensed and complex tannins, were evaluated for their cytotoxicities against human tumor cell lines, including malignant melanoma, lung carcinoma, ileocecal adenocarcinoma, epidermoid carcinoma, malignant melanoma, and medulloblastoma cell lines. Among them, chebulagic acid [1], geraniin [2], sanguiin H-11 [3], 4,5-di-O-galloylquinic acid [12], 1,3,4,5-tetra-O-galloylquinic acid [15], 1(beta)-O-galloylpedunculagin [24], furosin [29], castalagin [38], sanguiin H-2 [34], vescalagin [39], grandinin [40], phyllyraeoidin A [42], (-)-epicatechin 3-O-gallate [50], cinnamtannin B2 [55], and acutissimin A [56] exhibited moderate selective cytotoxicity against PRMI-7951 melanoma cells with ED50 values in the range of 0.1-0.8 microgram/ml. Selective cytotoxicities against the melanoma cells were also observed for strictinin [22], pedunculagin [23], eugeniin [25], elaeocarpusin [28], punicacortein C [37], casuarinin [41], sanguiin H-6 [43], procyanidin B-2 3,3'-di-O-gallate [51], procyanidin C-1 3,3',3"-tri-O-gallate [52], and cinnamtannin B1 [54] with ED50 values of 1-4 micrograms/ml. All of the tannins were found to be inactive (greater than 10 micrograms/ml) against lung carcinoma (A-549), ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of nasopharnyx (KB), and medulloblastoma (TE-671) tumor cells.

Antineoplastic Agents, Phytogenic↗

The effect of oral administration of polyethylene glycol on faecal helminth egg counts in pregnant goats grazed on browse containing condensed tannins.

Thirty yearling F1 Anglo-Nubian x Mubende goats, averaging 21 +/- 0.45 kg, kept on free-range feeding in the Ankole range land, Uganda, were screened for health and nutritional status, effectively treated against helminth parasites, mated, and randomly divided into two equal groups during a 3-month preparatory phase. During the 6 months that followed, the goats in one group received a daily oral dose (50 g/goat) of poly(ethylene glycol) (PEG), while the other group acted as the control (no PEG). The goats were monitored for faecal nematode egg counts and body weight gains, along with the quality of their diet, nematode contamination of the pasture, and the prevailing climatic factors in the area. Goats treated with PEG had significantly (p < 0.05) higher faecal helminth egg loads. The mean nematode eggs per gram of faeces (epg) of the PEG group (290 epg) was more than double that of the control group (129 epg). All the PEG-treated goats exhibited moderate to severe infections at the end of the experiment. The gain in body weight during gestation was lower (p < 0.05) in the PEG group (70.4 g per goat per day) than in the control group (91.8 g per goat per day). The PEG group lost 2.3 g per goat per day in the fifth month. PEG deactivates condensed tannins, and it was concluded that condensed tannins play a significant role in reducing the negative effects of gastrointestinal helminth burdens in the natural free-range feeding system of the Ankole range land in Uganda. Selective feeding on such range lands might expose goats to optimal concentrations of dietary condensed tannins with resultant beneficial effects.

Administration, Oral↗

Cultivar and processing effects on the pasting characteristics, tannin content and protein quality and digestibility of cowpea (Vigna unguiculata).

Four popular West African local cultivars of cowpea (Vigna unguiculata), with distinctly different seed coat colors, were evaluated for their relative amylograph pasting characteristics, condensed tannin content, in vitro protein digestibility and Tetrahymena protein efficiency ratio (t-PER). The effects of roasting and dehulling on these properties were also determined. There were wide variations in the hot paste viscosity characteristics of the different cultivars studied. The raw cowpea flour samples exhibited maximum paste viscosities ranging between 260 Brabender Units (BU) for the Mottled cultivar and 460 BU for the cream-colored Blackeye cultivar. Cowpea cultivars with the greatest peak viscosities showed low stabilities to extended cooking. Roasting depressed paste viscosity properties of all the cowpea cultivars studied. Tannin concentrations were 0.3-6.9 and 7.2-116 mg CE/g flour from whole cowpea seeds and seed coats respectively, increasing with intensity of seed color. Although dehulling removed 98% of the tannin content of raw cowpeas, improvement in protein quality as a result of dehulling was observed for only the highly-pigmented Maroon-red variety. Roasting significantly improved digestibility and more than doubled the t-PER of all cowpea cultivars studied. Roasted cowpeas possess adequate nutritional and functional qualities as protein supplements in cereal-based weaning foods. However, it appears that dehulling is necessary to enhance the nutritional quality of the highly pigmented cultivars of cowpea.

Amino Acids↗

Influence of Myriophyllum spicatum-derived tannins on gut microbiota of its herbivore Acentria ephemerella.

The submerged living larvae of Acentria ephemerella were fed in the laboratory with either M. spicatum or Potamogeton perfoliatus, two of their host plants. Larvae exhibited a reduced growth when fed M. spicatum, a freshwater angiosperm that contains high concentrations of tannins, secondary metabolites known for their herbivore-deterrent and antimicrobial properties. In this study, we investigated the influence of food-derived tannins on gut microbiota. Bacterial densities in the guts did not differ between the food regimes, ranging from 2.8 to 13.3 x 10(6) cells per gut. Gut bacteria were characterized with cultivation techniques and subsequent identification of the strains by molecular methods. We isolated 17 bacterial strains belonging to all subdivisions, i.e., we identified alpha-, beta-, and gamma-proteobacteria, Cytophyaga/Flavobacteria (CF) and several Gram-positive bacteria. All except one Gram-positive strain were found in the guts of larvae fed with P. perfoliatus. Gram-positive bacteria and bacteria of the CF cluster were more sensitive to polyphenol-containing extracts of M. spicatum in an agar diffusion assay than strains of the alpha- or gamma-proteobacteria subdivision. Our results suggest an influence of food-derived tannins on gut microbiota in A. ephemerella.

Adaptation, Physiological↗

Molecular analysis of herbivore-induced condensed tannin synthesis: cloning and expression of dihydroflavonol reductase from trembling aspen (Populus tremuloides).

In order to study condensed tannin synthesis and its induction by herbivory, a dihydroflavonol reductase (DFR) cDNA was isolated from trembling aspen (Populus tremuloides). Bacterial overexpression demonstrated that this cDNA encodes a functional DFR enzyme, and Southern analysis revealed that DFR likely is a single-copy gene in the aspen genome. Aspen plants that were mechanically wounded showed a dramatic increase in DFR expression after 24 h in both wounded leaves and unwounded leaves on wounded trees. Feeding by forest tent caterpillar (Malacosoma disstria) and satin moth (Leucoma salicis) larvae, and treatment with methyl jasmonate, all strongly induced DFR expression. DFR enzyme activity was also induced in wounded aspen leaves, and phytochemical assays revealed that condensed tannin concentrations significantly increased in wounded and systemic leaves. The expression of other genes involved in the phenylpropanoid pathway were also induced by wounding. Our findings suggest that the induction of condensed tannins, compounds known to be important for defense against herbivores, is mediated by increased expression of DFR and other phenylpropanoid genes.

Alcohol Oxidoreductases↗

Acceptance and digestibility of some selected browse feeds with varying tannin content as supplements in sheep nutrition in west Africa.

In 1994 and 1995 leaves from 10 selected West African trees and shrubs with varying tannin content were tested to determine their suitability as an alternative and supplementary browse feed for West African dwarf sheep to improve productivity in small-scale holdings in Benin. Dry matter intake per kg metabolic body weight (DM g/kg W0.75) varied between the different browse feeds and between the different trials and ranged from zero (Leucaena leucocephala) up to 26.7 DM g/kg W0.75 (Margaritaria discoidea). The digestibility of the organic matter varied between 58.9% (L. leucocephala) and 68.2% (Mallotus oppositifolius). Agelaea obliqua showed the highest levels of total phenols (10.2%), tannin phenols (8.8%) and extractable condensed tannins (8.0%). Leaves from various browse feeds are a good and protein-rich supplementary fodder in addition to the grass Panicum maximum. However, feeding of A. obliqua and Cnestis ferruginea should be avoided due to toxic components.

Animal Feed↗

The potential of wattle tannin extracts for fine use.

As a contribution to the fine use of wattle tannin, four fractions, purified tannin (Tp), ether extracts (Te), ethyl acetate extracts (Ta) and water fraction (Tw), were obtained by the methods of precipitation and liquid-liquid extraction of commercial wattle tannin. The number average molecular weights (M(n)) of Tp, Te, Ta and Tw were determined as 2660, 440, 980 and 2050 Da, respectively. The biological activities including protein precipitation, enzyme inhibition and antibacterial potency of each fraction were studied and compared. The results indicated that the molecular weight of each fraction is directly associated with its biological activities. The fraction of Tw showed the strongest capacity to precipitate protein and Ta showed the strongest capacity to inhibit enzyme activities, while Ta and Te showed better antibacterial potency.

Anti-Bacterial Agents↗