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Inhibitory effects of polyphenols in leaves of Artemisia princeps PAMP on protein fragmentation by Cu(II)-H2O2 in vitro.

The leaves of Artemisia princeps PAMP have traditionally been used as teas and foods in Japan. Polyphenols in Artemisia plants have been shown to have inhibitory effects against biological damages. The inhibitory effects of polyphenols in the leaves of A. princeps PAMP were investigated on protein fragmentation induced by Cu(II)-H(2)O(2) in vitro. The total polyphenol content in the leaves of A. princeps PAMP was 4.58%. The condensed tannin content was 0.62% by vanillin assay and 0.14% by proanthrocyanidin assay. The polyphenols in the leaves of A. princeps PAMP inhibited bovine albumin fragmentation by Cu(II)-H(2)O(2). The effects of polyphenols in the leaves of A. princeps PAMP were similar to those of tannic acid, studied as a related polyphenol. These results demonstrated that the leaves of A. princeps PAMP have inhibitory effects on protein fragmentation damage.

Antioxidants↗

Antioxidative effects of polyphenols in leaves of Houttuynia cordata on protein fragmentation by copper-hydrogen peroxide in vitro.

The leaves of Houttuynia cordata have been traditionally used as medicinal foods in East Asia. However, there have been few data about the biological effects. The antioxidative effects of polyphenols in the leaves of H. cordata were investigated on protein fragmentation by copper-hydrogen peroxide in vitro. The total polyphenol content in the leaves of H. cordata was 1.14%. The condensed tannin content was 2.46% by vanillin assay and 0.54% by proanthrocyanidin assay. The polyphenols in the leaves of H. cordata inhibited bovine serum albumin fragmentation by copper-hydrogen peroxide. These results demonstrated that the leaves of H. cordata have antioxidative effects on biological damage such as protein fragmentation.

Antioxidants↗

Glandular hairs of Sigesbeckia jorullensis Kunth (Asteraceae): morphology, histochemistry and composition of essential oil.

Long-stalked glandular hairs of outer and inner involucral bracts of Sigesbeckia jorullensis, which are important for epizoic fruit propagation, were investigated using light and scanning electron microscopy. The essential oil secreted by the hairs was analysed by chromatographic methods including gas chromatography/mass spectrometry and with a laser microprobe mass analyser. The glandular hairs consisted of a large multicellular stalk and a multicellular secreting head. The apical layer of glandular head cells was characterized by leucoplasts and calcium oxalate crystals. Below the apical cells there were up to six layers of cells containing many chloroplasts around the nucleus and surrounded by vacuoles filled with flavonoids and tannins. The essential oil originating in the head cells was secreted into the subcuticular space and may be liberated by rupture of the cuticle. It was mainly composed of sesqui- and diterpenes, with the sesquiterpene hydrocarbon germacrene-D as the main component. Monoterpenes, n-alkanes and their derivatives as well as flavonoid aglycones were also detected. The stickiness of the essential oil is probably associated with the high content of oxygenated sesqui- and diterpenes. In addition to long-stalked trichomes, small biseriate trichomes occurred, secreting small quantities of essential oil into a subcuticular space.

Asteraceae↗

Specificity of glossopharyngeal nerve responses to astringent compounds in Xenopus.

Astringent compounds were applied to oral epithelium of the clawed toad, Xenopus laevis, and rapidly rising and highly sensitive responses could be recorded from the whole glossopharyngeal nerve, but not at all from the trigeminal nerve. The response to 10 mM tannic acid decreased progressively with repetitive application. These responses to tannic acid, however, recovered completely by treating with chemicals capable of forming strong hydrogen and hydrophobic bonds. These chemical bondings are generally recognized as a model for polyphenol (tannin)-protein interactions based on physico-chemical measurements in vitro. The high affinities of these chemicals for tannic acid may be effective in releasing both bonds in the interaction of tannic acid with the receptor molecules. Our results provide in vivo evidence for this model.

Animals↗

Increased efficacy of Bacillus thuringiensis subsp. kurstaki in combination with tannic acid.

We identified tannic acid as an inexpensive additive that increased the efficacy of sublethal concentrations of Bacillus thuringiensis subsp. kurstaki (Berliner). Tannic acid mimicked the active constituents contained in an aqueous, tannin-rich extract of Taxus baccata (L.) bark that retarded development of Heliothis virescens (F.) larvae at 10,000 ppm; most larvae remained in first and second stage when treated with 250-10,000 ppm of tannic acid. Instar development of Trichoplusia ni (Hübner) larvae was affected in a concentration-dependent manner by 2.5-500 ppm of tannic acid. In subsequent bioassays, tannic acid at 25-500 ppm in combination with B. thuringiensis (1.63 micrograms [AI]/ml diet) yielded mean mortalities of 57-75%, whereas treatments with B. thuringiensis alone produced 10% mortality. Mean mortalities in the 3.0, 4.5, and 6.75 micrograms (AI) B. thuringiensis per milliliter of diet treatments (5.5; 8.0, and 30%, respectively) were significantly higher in the presence of 250 and 2,500 ppm tannic acid; in these treatments we observed 78-94% mortality. Addition of tannic acid increased the activity of concentrations of 3-4.5 micrograms (AI) B. thuringiensis per milliliter of diet to approximately that of a concentration of 13 micrograms (AI) B. thuringiensis per milliliter of diet alone (85-95% mortality). Although deaths caused by a formulation of B. thuringiensis + tannic acid occurred more slowly than with high rates of B. thuringiensis alone, such formulations would have the advantages of arresting development, minimizing foliar damage, and decreasing the concentration of B. thuringiensis used.

Animals↗

Plant allocation to defensive compounds: interactions between elevated CO(2) and nitrogen in transgenic cotton plants.

Plant allocation to defensive compounds in response to growth in elevated atmospheric CO(2) in combination with two levels of nitrogen was examined. The aim was to discover if allocation patterns of transgenic plants containing genes for defensive chemicals which had not evolved in the species would respond as predicted by the Carbon Nutrient Balance (CNB) hypothesis. Cotton plants (Gossypium hirsutum L.) were sown inside 12 environmental chambers. Six of them were maintained at an elevated CO(2) level of 900 micromol mol(-1) and the other six at the current level of approximately 370 micromol mol(-1). Half the plants in each chamber were from a transgenic line producing Bacillus thuringiensis (Bt) toxin and the others were from a near isogenic line without the Bt gene. The allocation to total phenolics, condensed tannins, and gossypol and related terpenoid aldehydes was measured. All the treatments were bioassayed against a non-target insect herbivore found on cotton, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Plants had lower N concentrations and higher C:N ratios when grown in elevated CO(2). Carbon defensive compounds increased in elevated CO(2), low N availability or both. The increase in these compounds in elevated CO(2) and low N, adversely affected growth and survival of S. exigua. The production of the nitrogen-based toxin was affected by an interaction between CO(2) and N; elevated CO(2) decreased N allocation to Bt, but the reduction was largely alleviated by the addition of nitrogen. The CNB hypothesis accurately predicted only some of the results, and may require revision. These data indicate that for the future expected elevated CO(2) concentrations, plant allocation to defensive compounds will be affected enough to impact plant-herbivore interactions.

Adaptation, Physiological↗

Low nutritional quality of unconventional tropical crop seeds in rats.

As the search for alternative sources of food to alleviate hunger continues, this study was undertaken to determine the biological value in growing rats (BV) of proteins of some lesser known tropical seeds gathered in Nigeria. Antinutritional factors (trypsin inhibitors, phytic acid, oxalate, tannin, alkaloids) and amino acid compositions were also determined, and protein digestibility-corrected amino acid score (PDCAAS) was calculated using the amino acid requirement pattern of the preschool child and individual seed-specific correction factors for crude protein. A rat growth and balance study was conducted to determine digestibility, nitrogen-, and energy balance by feeding as the only unsupplemented protein source milled and heat-treated seeds of Adansonia digitata (Bombacaceae) and Prosopis africana, Lonchocarpus sericeus, Enterolobium cyclocarpium, Sesbania pachycarpa and Pterocarpus osun (Leguminosae) in comparison to casein fortified with methionine (control). Diets containing P. africana and L. sericeus seeds caused poor feed intake and weight loss in rats and were excluded from the nitrogen-balance test. Among the seed samples, S. pachycarpa followed by A. digitata showed the most advantageous nutritional quality [amino acid composition, digestibility, BV and net protein utilization (NPU)]. True digestibility was 82.9 and 74.5 vs. 98.5, BV was 64.6 and 70.0 vs. 90.4, and NPU was 53.5 and 52.1 vs. 89.0 for S. pachycarpa and A. digitata vs. casein (control), respectively. In terms of PDCAAS, lysine was the first limiting amino acid for S. pachycarpa (88%) and for A. digitata (58%). The PDCAAS of all essential amino acids was below 100% for E. cyclocarpium (e.g., cysteine + methionine: 37%) and for P. africana (e.g., threonine: 46%, except valine and a very high content of cysteine and methionine). In conclusion, all seeds tested in the rat balance trial were of inferior quality compared to casein. Before these tropical seeds could be used as food components or feed supplements, safety studies and proper processing to remove antinutritional factors and possible toxic constituents were required.

Alkaloids↗

Ileal endogenous nitrogen recovery is increased and its amino acid pattern is altered in pigs fed quebracho extract.

Ileal endogenous nitrogen recovery (ENR) in pigs (9 +/- 0.6 kg body weight) was estimated simultaneously using the (15)N-isotope dilution technique ((15)N-IDT) and the peptide alimentation ultrafiltration (UF) method. Diets were cornstarch, enzyme-hydrolyzed casein with no (control) or high (4%) content of quebracho extract (Schinopsis spp.) rich in condensed tannins. The amino acid (AA) pattern of the ENR was also determined. The ENR of pigs fed the quebracho diet was higher (P = 0.0001) than that of pigs fed the control diet [6.00 vs. 1.95 g/kg dry matter intake (DMI) for the (15)N-IDT and 5.18 vs. 1.49 g/kg DMI for the UF method, respectively]. With the (15)N-IDT, ENR values were 0.44-0.79 g/kg DMI (24%) higher (control P = 0.0032, quebracho P = 0.0002) than for the UF method. Apparent nitrogen digestibility depended on diet (69.0% quebracho vs. 86.0% control, P = 0.0001). Real nitrogen digestibility (RD-N) determined by the UF method was higher (P = 0.0001) for the control than for the quebracho diet (91.4 vs. 88.2%). Corresponding values for the (15)N-IDT did not differ (P = 0.0569) between diets (92.8 vs. 91.4%). The (15)N-IDT gave higher values for RD-N of both diets (control P = 0.0030, quebracho P = 0.0002) compared with the UF method. Endogenous AA recoveries (g/kg DMI) were increased 300% (P = 0.0001) and the AA-pattern of ENR was changed (P from 0.0001 to 0.7530 for different AA) by the quebracho diet. A constant AA-pattern of ENR cannot be assumed. Despite limitations of both techniques, the (15)N-IDT and the UF method gave similar results with respect to ENR.

Amino Acids↗

Carcinogenicity of some folk medicinal herbs in rats.

Twelve medicinal herbs were bioassayed to correlate a high incidence of esophageal carcinoma in natives of different places with their habitual consumption of these products. Outbred NIH Black rats were given 72 weekly sc injections of the total aqueous extracts of the plant materials. The tanninrich plant extracts from Areca catechu and Rhus copallina produced local tumors in 100 and 33%, respectively, of the experimental animals. Other materials included Diospyros virginiana and extracts from plants not rich in tannins. Diospyros and extracts of Sassafras albidum and Chenopodium ambrosiodes were tumorigenic in over 50% of the treated animals.

Animals↗

Moderate inhibition of mutagenicity and carcinogenicity of benzo[a]pyrene, 1,6-dinitropyrene and 3,9-dinitrofluoranthene by Chinese medicinal herbs.

The activity of six Chinese medicinal herbs against the environmental mutagens and carcinogens benzo[a]pyrene (B[a]P), 1,6-dinitropyrene (1,6-diNP) and 3,9-dinitrofluoranthene (3,9-diNF) was determined. Samples of Prunella spica, Rheum palmatum, Polygonum multiflorum, Agrimonia pilosa, Ephedra sinica and Teitoutou were tested in an in vitro system. Antimutagenic activity against B[a]P was marked in the presence of extracts (boiled for 2 h in a water bath) whereas that against 1,6-diNP and 3,9-diNF varied from 20 to 86%. The differences in inhibition might be due to inactivation of metabolic enzymes. An extract of P. multiflorum was divided into ether, ethyl acetate and water soluble fractions, which were tested for antimutagenic activity against B[a]P. The antimutagenic action of the ethyl acetate soluble fraction was substantial and dose-dependent. Tannins and related compounds were the major components of the extract, of which epigallocatechin, epigallocatechin gallate, epicatechin gallate and tannic acid strongly inhibited the mutagenicity of B[a]P (2.5 micrograms/plate) in Salmonella typhimurium TA98 with S9 mix. To confirm the results of the in vitro test system, F344/DuCrj male rats were given a subcutaneous injection of B[a]P. Thereafter, they received water extracts of the six Chinese medicinal herbs for 50 weeks and were examined for tumors. The P. multiflorum extract significantly reduced the tumor incidence.

Animals↗

Induction of cell cycle arrest and apoptosis in human non-small cell lung cancer A549 cells by casuarinin from the bark of Terminalia arjuna Linn.

Casuarinin, a hydrolyzable tannin isolated from the bark of Terminalia arjuna Linn. (Combretaceae), inhibits human non-small cell lung cancer A549 cells by blocking cell cycle progression in the G0/G1 phase and inducing apoptosis. Enzyme-linked immunosorbent assay showed that the G0/G1 phase arrest is due to p53-dependent induction of p21/WAF1. An enhancement in Fas/APO-1 and the two forms of Fas ligand (FasL), membrane-bound FasL and soluble FasL, might be responsible for the apoptotic effect induced by casuarinin. Our study reports here for the first time that the induction of p53 and the activity of the Fas/FasL apoptotic system may participate in the antiproliferative activity of casuarinin in A549 cells.

Antineoplastic Agents, Phytogenic↗

Inhibition of HIV-1 replication by an aqueous extract of Spirulina platensis (Arthrospira platensis).

An aqueous extract of the blue-green filamentous algae Arthrospira platensis (previously called Spirulina platensis) inhibited HIV-1 replication in human T-cell lines, peripheral blood mononuclear cells (PBMC), and Langerhans cells (LC). Extract concentrations ranging between 0.3 and 1.2 microg/ml reduced viral production by approximately 50% (50% effective concentration [EC50]) in PBMCs. The 50% inhibitory concentration (IC50) of extract for PBMC growth ranged between 0.8 and 3.1 mg/ml. Depending on the cell type used, therapeutic indices ranged between 200 and 6000. The extract inactivated HIV-1 infectivity directly when preincubated with virus before addition to human T-cell lines. Fractionation of the extract revealed antiviral activity in the polysaccharide fraction and also in a fraction depleted of polysaccharides and tannins. We conclude that aqueous A platensis extracts contain antiretroviral activity that may be of potential clinical interest.

Cell Line↗

Papillibacter cinnamivorans gen. nov., sp. nov., a cinnamate-transforming bacterium from a shea cake digester.

A new, strictly anaerobic, Gram-positive, non-sporulating, mesophilic bacterium, designated strain CIN1T (T=type strain) was isolated from an anaerobic digester fed with shea cake rich in tannins and aromatic compounds. Cells of strain CIN1T were rod-shaped, had characteristically pointed ends (1.3-3.0 x 0.5-0.6 microm) and occurred singly, in pairs and sometimes in chains of up to six. The pH range for growth was 6.9-8.5 and the temperature growth range was 15-40 degrees C. Optimum growth occurred with yeast extract and cinnamate at 37 degrees C and a pH of 7.5. The isolate transformed cinnamate by degrading the aliphatic side chain to produce acetate and benzoate rather than by aromatic ring cleavage or demethoxylation. The position of the methoxyl group appears to be important in the degradation of the aliphatic side chain of cinnamate; consequently, 3-methoxycinnamate and 4-methoxycinnamate, but not 2-methoxycinnamate, are transformed to produce acetate and methoxybenzoates, namely 3-methoxybenzoate and 4-methoxybenzoate, respectively. Crotonate is degraded to acetate and butyrate. The G+C content of the DNA is 56 mol%. Phylogenetic analysis of the 16S rRNA gene of strain CIN1T indicated that it was a member of the low-G+C-containing Gram-positive branch with a specific relationship to Sporobacter termitidis (sequence identity of 88%). The phylogenetic results concur with the phenotypic data which reveals that the isolate is a novel bacterium and, based on these findings, strain CIN1T (= DSM 12816T = ATCC 700879T) has been designated Papillibacter cinnamivorans gen. nov., sp. nov.

Anaerobiosis↗

Induction of biomolecules in mature leaves of Terminalia arjuna due to feeding of Antheraea mylitta Drury.

Terminalia arjuna is an important food plant of the tasar silkworm, Antheraea mylitta Drury. In this study, we investigated the induction of biomolecules in mature leaves of these plants subjected to insect feeding. Increase in total tannin content, lipid peroxidation, and trypsin inhibitor activity have been observed in mature leaves damaged by the insects. The growth rate of Vth instar larvae of A. mylitta fed on previously damaged foliage reduced by 87.1%. Induction of biomolecules for defense mechanisms in relation to herbivore damage has been discussed.

Animals↗

The effect of environmental factors on the distribution of Neoparamoeba pemaquidensis in Tasmania.

Aquaculture in Tasmania is mostly carried out in estuaries. These estuarine habitats show a great variety and form unique environments in which Neoparamoeba pemaquidensis, the amoebic gill disease (AGD)-causing protozoan, may or may not survive. Tasmania is divided into two zones, one where AGD is present and one where AGD is absent, but any ecological data to rationalize this distribution is lacking. In in vitro trials N. pemaquidensis strains were exposed to different concentrations of ammonium sulphate, copper sulphate, copper sulphate and tannin, and different Neoparamoeba densities, salinities and temperatures. A trial using field water samples investigated the survival of N. pemaquidensis in waters sourced from AGD-free and AGD-positive zones, and water analysis was performed to determine any differences. Significantly decreased protozoan survival was found with exposure to increasing copper sulphate concentrations from 10 to 100,000 microM (P < 0.001), salinity of 15 per thousand (P < 0.001), low Neoparamoeba densities of 625 and 1,250 cells mL(-1) (P = 0.0005), and water sourced from Macquarie Harbour (P < 0.001). The water chemistry of this AGD-free zone showed significantly lower dissolved calcium and magnesium concentrations which may contribute to this area being AGD-free. Understanding of the ecology of N. pemaquidensis will enable better control and prevention strategies for Tasmanian salmon growers.

Ammonium Sulfate↗

The morphology of echinoid phagocytes and mouse peritoneal macrophages during phagocytosis in vitro.

The morphology of mouse peritoneal macrophages and echinoid phagocytes during phagocytosis in vitro was studied. A striking similarity in the function of the foreign surface receptor is found in the two systems. Glutaraldehyde-treated erythrocytes attached randomly over the entire surface of the cells and were internalized without circumferential attachment between the particles and the phagocyte membrane. The particles seemed to sink directly into the cytoplasm of the cells. Tannin-treated erythrocytes were phagocytosed by the echinoid cells in a similar mode. The complement-coated erythrocytes were attached only in the perinuclear area of the echinoid phagocyte's membrane, but the morphology of their internalization was similar to that mediated by the foreign surface receptor. A circumferential attachment between the particles and the phagocyte membrane did not seem necessary. This is also the case for mouse peritoneal macrophages.

Animals↗

Protection of spermatogenesis in mice against gamma ray induced damage by Hippophae rhamnoides.

The radioprotective action of a preparation from Hippophae rhamnoides berries RH-3, already reported to render >80% survival against whole body 10 Gy gamma irradiation, was further investigated with respect to the testicular system. RH-3 was administered to mice 30 min before gamma irradiation (5 and 10 Gy) and histological parameters such as testis weight, sperm count, frequency of abnormal sperm, repopulation index, stem cell survival index and seminiferous tubular diameter were assessed on the 35th day. RH-3 administration partially countered radiation induced reduction in testis weight, sperm count, repopulation index and stem cell survival index (p < 0.01). The increase in the frequency of abnormal sperm (15.17 +/- 1.046%) caused by irradiation (5 Gy) was counteracted by pre-irradiation treatment with RH-3, which significantly decreased the level of abnormal spermatozoa to 7.99 +/- 0.918% (p < 0.001), i.e. 52% abnormalities in comparison with 5 Gy irradiated group. RH-3 treatment alone did not elicit any toxic or adverse effect on the process of spermatogenesis. The present study suggests that RH-3 treatment protected spermatogenesis by enhancing the spermatogonial proliferation, enhancing the stem cell survival and reducing sperm abnormalities. The presence of polyphenolic flavonoids and tannins in the extract and the radical scavenging activity might be responsible for the radioprotective action of RH-3.

Animals↗

Evidence of species interactions within an ectomycorrhizal fungal community.

Ectomycorrhizal fungal communities can be structured by abiotic and biotic factors. Here, we present evidence for community structuring by species interactions. We sampled ectomycorrhizas and forest floor seven times during a 13-month period. The presence of various ectomycorrhizal fungal species was determined for each sample, and species co-occurrence analyses were performed. For both ectomycorrhizas and forest floor samples there was significantly less co-occurrence among species within the community than expected by chance, mostly because of negative associations involving Cenococcum geophilum or Clavulina cinerea. For some species pairs, there was significantly more co-occurrence than expected by chance. Both nitrogen and tannin additions to the forest floor altered some interactions among species. The causes of these nonrandom distributions are currently unknown. Future investigations on competition, antibiosis, parasitism and facilitation among ectomycorrhizal fungal species appear to be warranted.

Hyphae↗