Aminomethylphosphonic acid, a metabolite of glyphosate, causes injury in glyphosate-treated, glyphosate-resistant soybean

Glyphosate-resistant (GR) soybean [Glycine max (L.) Merr.] was developed by stable integration of a foreign gene that codes insensitive enzyme 5-enolpyruvylshikimate-3-phosphate synthase, an enzyme in the shikimate pathway, the target pathway of glyphosate. Application of glyphosate to GR soybean results in injury under certain conditions. It was hypothesized that if GR soybean is completely resistant to the glyphosate, injury could be caused by a metabolite of glyphosate, aminomethylphosphonic acid (AMPA), a known phytotoxin. Glyphosate and AMPA effects on one- to two-trifoliolate leaf stage (16-18-days old) GR and non-GR soybean were examined in the greenhouse. In GR soybean, a single application of glyphosate-isopropylammonium (1.12-13.44 kg/ha) with 0.5% Tween 20 did not significantly reduce the chlorophyll content of the second trifoliolate leaf at 7 days after treatment (DAT) or the shoot dry weight at 14 DAT compared with Tween 20 alone. A single application of AMPA (0.12-8.0 kg/ha) with 0.5% Tween 20 reduced the chlorophyll content of the second trifoliolate leaf by 0-52% at 4 DAT and reduced shoot fresh weight by 0-42% at 14 DAT in both GR and non-GR soybeans compared with Tween 20 alone. AMPA at 0.12 and 0.50 kg/ha produced injury in GR and non-GR soybean, respectively, similar to that caused by glyphosate-isopropylammonium at 13.44 kg/ha in GR soybean. AMPA levels found in AMPA-treated soybean of both types and in glyphosate-treated GR soybean correlated similarly with phytotoxicity. These results suggest that soybean injury to GR soybean from glyphosate is due to AMPA formed from glyphosate degradation.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.     

Protocatechuic acid, a metabolite of anthocyanins, inhibits monocyte adhesion and reduces atherosclerosis in apolipoprotein E-deficient mice

Polyphenols, including anthocyanins, from various plant foods are effective in the prevention of atherosclerosis in animal and human studies. Protocatechuic acid (PCA), a major metabolite of anthocyanins, has been found to possess the anti-carcinogenic effect, whereas the in vivo effect of PCA as an anti-atherosclerotic agent remains unknown. We demonstrated herein that PCA inhibited monocyte adhesion to tumor necrosis factor-α (TNF-α)-activated mouse aortic endothelial cells, associated with the inhibition of vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) expression. Furthermore, PCA inhibited the nuclear content of p65, a subunit of nuclear factor-κB (NF-κB), along with reduced NF-κB binding activity. Finally, PCA administration in the apolipoprotein E (ApoE)-deficient mouse model reduced aortic VCAM-1 and ICAM-1 expression, NF-κB activity, and plasma-soluble VCAM-1 and ICAM-1 levels, with inhibiting atherosclerosis development. We suggest that PCA possesses the anti-atherogenic effect at least partially via its anti-inflammatory activity.     

Nutritive and antioxidative potential of fresh and stored pomegranate industrial byproduct as a novel beef cattle feed

Pomegranate peel is a nutritive-rich byproduct whose amounts are extensively growing due to the exponential increase in the production of pomegranate juice and "ready to eat" arils. Pomegranate peel is a rich source for antioxidants and thus may serve in the prevention of cattle diseases and in the improvement of beef products, making it an attractive component in beef cattle diets. The present study aims to evaluate the effect of commonly used storage practices on the nutritive and antioxidative properties of pomegranate peel. In general, storage conditions preserved most antioxidant capacity. Ensiling ambivalently affected the nutritive values of the peel and promoted increased levels of antioxidative components. In addition to polyphenols, nonphenolic components, such as alpha- and gamma-tocopherols, contributed to the total antioxidative capacity, and several minerals found in the peel added to its nutritional value. Dietary supplementation with fresh peels promoted significant increases in feed intake and alpha-tocopherol concentration in the plasma, with positive tendency toward increased weight gain of bull calves. All in all, the nutritive value and the antioxidant capacity of pomegranate peel turn it into a favorable health-promoting constituent of feedlot beef cattle diet.     

1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 1,2, 3,4-tetrahydro-beta-carboline-3-carboxylic acid in fruits

1,2,3,4-Tetrahydro-beta-carboline-3-carboxylic acid (THCA) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA), as two diastereoisomers (1S,3S and 1R,3S), occurred in commercial fruits. Citrus fruits exhibited the highest content; other fruits contained very low levels or none at all. The content of MTCA was as follows: orange, 0.35-2.47 microg/g; lemon, 0.15-2.05 microg/g; grapefruit, 1.12-8.37 microg/g; mandarin, 0.57-2.5 microg/g; banana, nd-0.74 microg/g; pear, nd-0.017 microg/g; grape, 0.01-0.22 microg/g, tomato, 0.05-0.25 microg/g; and apple, nd-0.012 microg/g). THCA, if present, usually occurred at <0.05 microg/g. Fruit ripening and softening during storage were accompanied with a significant increase of MTCA, in both pears and bananas. Those and previous results confirm that foods are an important source of tetrahydro-beta-carbolines in humans.     

Cynodontin: a fungal metabolite with antifungal properties

A red pigment that accumulates in cultures of a Drechslera avenae pathotype with specificity for Avena sterilis was isolated and identified as the anthraquinone cynodontin (3-methyl-1,4,5,8-tetrahydroxyanthraquinone). Satisfactory yield of the compound was obtained with 20-60 day incubations at temperatures between 20 and 27 degrees C. Cynodontin was tested in vitro for fungitoxicity and was found to be a potent inhibitor of the growth of Sclerotinia minor, Sclerotinia sclerotiorum, and Botrytis cinerea and, to a lesser extent, of Verticillium dahliae. The ED(50) values obtained with these fungi were of the same order of magnitude as those of the commercial fungicides dicloran and carbendazim, which were used as reference chemicals. In contrast, the growth of a number of other fungi was not significantly inhibited by cynodontin. Anthraquinone and two other anthraquinone derivatives, emodin and chrysophanol, which were also included in the tests, did not affect the growth of the cynodontin-sensitive fungi. It thus appears that the type and position of the substitutions at the C-ring play a role in the expression of antifungal activity.     

3-Mercapto-2-methylpentan-1-ol, a new powerful aroma compound

3-Mercapto-2-methylpentan-1-ol was first detected in a complex thermally processed flavor and finally isolated from raw onions. The chemical structure of this new compound was identified by MS and (1)H NMR measurement and synthesis of the proposed structure. Sensory evaluation at different concentrations indicated that the flavor quality is strongly dependent on concentration. At low concentration (0.5 ppb) a pleasant meat broth, sweaty, onion, and leek-like odor can be perceived. On the basis of some isolation experiments and volatiles occurring in raw onions, a formation pathway is proposed. As one intermediate 3-mercapto-2-methylpentanal, another new strong flavor compound, was suggested. The presence of this compound in raw onions was confirmed by synthesis and comparison of MS and chromatographic data.     

Identification, quantitative determination, and antioxidative activities of chlorogenic acid isomers in prune (Prunus domestica L. )

Neochlorogenic acid (3-CQA) and cryptochlorogenic acid (4-CQA), isolated from prune (Prunus domestica L.), were identified by NMR and MS analyses. In addition, the quantity of chlorogenic acid isomers in prune were measured by HPLC. These isomers, 3-CQA, 4-CQA, and chlorogenic acid (5-CQA), were contained in the ratio 78.7:18. 4:3.9, respectively. 4-CQA was identified and quantified in prune for the first time, and relatively high amounts of this isomer were characteristic. Antioxidative activities of the chlorogenic acid isomers, such as scavenging activity on superoxide anion radicals and inhibitory effect against oxidation of methyl linoleate, were also evaluated. Each isomer showed antioxidative activities which were almost the same.     

Evaluation of antioxidative activity of extracts from a brown seaweed,Sargassum siliquastrum

Antioxidative activities of the extracts from Sargassum siliquastrum were determined using the inhibition of red blood cell (RBC) hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radicals, suppression of lipid peroxidation using rat brain homogenate, and scavenging activity of superoxide radicals. The dichloromethane fraction isolated from the methanol crude extract by differential solvent extractions exhibited the strongest antioxidant activity in both RBC hemolysis and lipid peroxidation assays. This fraction was further fractionated into four subfractions F1-F4 by silica gel column chromatography. F1 was found to be most effective in protecting RBC against AAPH radicals and in inhibiting lipid peroxidation. On the basis of thin-layer chromatography and UV and IR spectra analyses, all subfractions contained phenolic compounds. However, there was no correlation between the above antioxidant potency and total phenolic compounds estimated by using the Folin-Ciocalteau method.     

Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds

Dietary ellagic acid and related polyphenols are metabolized in humans to dibenzopyran-6-one derivatives, and the microbial origin of these metabolites has been suggested. However, this has not been demonstrated so far. Fecal samples donated by six volunteers were incubated under anaerobic conditions, and aliquots were used to evaluate the fecal metabolism of ellagic acid, the ellagitannin punicalagin, and an ellagitannin rich extract from walnuts. The isoflavone daidzein was also incubated with the same fecal samples to follow the production of the microbial metabolites previously reported (dihydrogenistein, O-demethylangolensin, and equol) as a positive control of the system and to evaluate similarities between isoflavone and ellagic acid fecal flora metabolism. After fermentation the metabolite "urolithin A" (3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one) was produced from ellagic acid, punicalagin, and the ellagitannin extract in all the fecal cultures from different volunteers, but with very different production rates and concentrations. This large variability in the concentration of metabolite and kinetics of metabolite production is consistent with the large variability found in the excretion of these metabolites in urine in vivo after human consumption of ellagitannins, and with differences in the composition of the fecal microflora. No correlation between isoflavone and ellagic acid metabolism by fecal microflora was observed. The present study confirms the microbial origin of the recently reported in vivo generated hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives in humans and is a further step in the study of the bioavailability and metabolism of ellagic acid and ellagitannins.     

Isolation and structure determination of new antioxidative ferulic acid glucoside esters from the rhizome of Alpinia speciosa, a Zingiberaceae plant used in Okinawan food culture

An assay-guided isolation gave three antioxidants including two newly identified compounds from the rhizomes of Alpinia speciosa, which is used as an important plant in the food culture of the Okinawa area of Japan. Spectroscopic analysis of the two new compounds revealed them to be new glucoside esters of ferulic acid. The antioxidant activity of the esters was measured using two different methods. Both compounds showed greater activity than that of Trolox in the TLC method; however, one of the compounds showed weaker inhibitory activity than that of Trolox and epicatechin against AMVN-induced methyl linoleate oxidation.     

Synergistic antioxidative effects of alkamides, caffeic acid derivatives, and polysaccharide fractions from Echinacea purpurea on in vitro oxidation of human low-density lipoproteins

Preparations of Echinacea are widely used as alternative remedies to prevent the common cold and infections in the upper respiratory tract. After extraction, fractionation, and isolation, the antioxidant activity of three extracts, one alkamide fraction, four polysaccharide-containing fractions, and three caffeic acid derivatives from Echinacea purpurea root was evaluated by measuring their inhibition of in vitro Cu(II)-catalyzed oxidation of human low-density lipoprotein (LDL). The antioxidant activities of the isolated caffeic acid derivatives were compared to those of echinacoside, caffeic acid, and rosmarinic acid for reference. The order of antioxidant activity of the tested substances was cichoric acid > echinacoside > or = derivative II > or = caffeic acid > or = rosmarinic acid > derivative I. Among the extracts the 80% aqueous ethanolic extract exhibited a 10 times longer lag phase prolongation (LPP) than the 50% ethanolic extract, which in turn exhibited a longer LPP than the water extract. Following ion-exchange chromatography of the water extract, the majority of its antioxidant activity was found in the latest eluted fraction (H2O-acidic 3). The antioxidant activity of the tested Echinacea extracts, fractions, and isolated compounds was dose dependent. Synergistic antioxidant effects of Echinacea constituents were found when cichoric acid (major caffeic acid derivative in E. purpurea) or echinacoside (major caffeic acid derivative in Echinacea pallida and Echinacea angustifolia) were combined with a natural mixture of alkamides and/or a water extract containing the high molecular weight compounds. This contributes to the hypothesis that the physiologically beneficial effects of Echinacea are exerted by the multitude of constituents present in the preparations.     

Dicyandiamide enhances liming potential of two legume materials when incubated with an acid Ultisol

The incorporation of two legume materials increased soil pH, while the nitrification of the ammonium-nitrogen (N) from the ammonification of organic N weakened the liming potential of these materials at the later stages of incubation experiments. The addition of dicyandiamide (DCD) enhanced the liming potential of the two legume materials through its inhibition of nitrification of ammonium-N. At the end of the 60-d incubation, soil pH of the treatment using Chinese milk vetch shoots with DCD was 1.48 units higher than that of control and 1.16 units higher than of the treatment using only Chinese milk vetch. The corresponding data for the pea straw were 1.24 and 1.01 units higher, respectively.     

Rhizobium strain,a banana (Musa spp.)-associated bacterium with a high potential as biofertilizer

A diazotrophic bacterial strain denominated 11B isolated from the rhizosphere of a banana plant (Musa spp.) was characterized morphologically, biochemically, and phylogenetically, whereas the symbiotic potential of the strain was assessed through tests of host range, interstrain nodulation competitiveness, and capacity to synthesize indole-3-acetic acid (IAA) and solubilize phosphate. Based on morphological and physiological–biochemical properties, as well as 16S ribosomal deoxyribonucleic acid (rDNA) sequence and phylogenetic analysis, the strain 11B belonged to the genus Rhizobium with 100.0% sequence similarity with Rhizobium tropici CAF440. The optimum growth temperature and pH for strain 11B are 28°C and 7.2, respectively. This strain was able to produce IAA, solubilize phosphate, and fix large amounts of nitrogen (N₂) and form effective nodules on the legumes Acaciella angustissima, Gliricidia sepium, Leucaena leucocephala, Lysiloma acapulcensis, and Phaseolus vulgaris. The rhizobial strain 11B was used successfully as a biofertilizer in agriculturally important legumes, forest trees, and agroindustrial plants.     

Identification of 4-methylspinaceamine, a pictet-spengler condensation reaction product of histamine with acetaldehyde, in fermented foods and its metabolite in human urine

Previous study demonstrated that 4-methylspinaceamine (4-methyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine), a Pictet-Spengler condensation reaction product of histamine with acetaldehyde, is present in human urine. The current study sought to determine whether 4-methylspinaceamine is present in fermented foods; its presence might be expected since both histamine and acetaldehyde are often present in these foods. Soy sauce, fish sauce, cheese, and shao hsing wine (Chinese wine) were found to contain 4-methylspinaceamine. The concentration of 4-methylspinaceamine excreted in human urine was greatly elevated after ingestion of a meal containing soy sauce as a dietary source of 4-methylspinaceamine, demonstrating that the level of 4-methylspinaceamine in human urine was affected by dietary foods. In addition, a metabolite of 4-methylspinaceamine in human urine was investigated. An enhanced peak in the HPLC chromatogram of human urine samples after ingestion of 4-methylspinaceamine-containing foods was observed. A peak at the same retention time was also observed from a human urine sample after administration of 4-methylspinaceamine, suggesting that the peak was due to a metabolite. By comparison with the newly synthesized authentic compound, the metabolite was identified as 1,4-dimethylspinaceamine.     

Enzymatic synthesis of 4-amino-3,5-diethylphenyl sulfate, a rodent metabolite of alachlor.

Rat liver tissue homogenates were utilized for in vitro enzymatic conversion of 2,6-diethylaniline (DEA) to the important alachlor metabolite 4-amino-3,5-diethylphenyl sulfate (ADEPS), which was also generated as a radiolabeled standard for use in metabolism studies. ADEPS formation in rodents is associated with the production of other reactive metabolites implicated in alachlor rodent carcinogenesis, making dependable access to an ADEPS standard highly desirable. (14)C-DEA was oxidized by rat liver microsomes to (14)C-4-amino-3,5-diethylphenol, which was further converted to ADEPS via addition of the phosphosulfate transferase cofactor adenosine-3'-phosphate-5'-phosphosulfate. Microprobe NMR was used in conjunction with high-resolution mass spectrometry to fully characterize the resulting (14)C-ADEPS and confirm its structure. Because microgram quantities sufficed for full characterization, the enzymatic transformation provides a viable alternative to radiosynthesis of (14)C-ADEPS.     

The potential acidity associated with dews,frosts, and fogs

Acidity associated with special meteorological events such as dews, frosts, and fogs may cause significant environmental effects ranging from increased leaching of plant metabolites to material corrosion. In addition, dews, frosts and fogs occur with much greater frequencies than rain or snow in most areas. Due to these facts, a theoretical study was conducted to determine the potential acidity associated with these events. Results from this study showed the potential acidity associated with these events to be significant. Values of fog pH are extremely sparse, but indicate that they are slightly acidic. However, these experiments have probably under-estimated the actual acidic potential of dew and frost. Theoretical calculations indicate that synergistic acidity from dew or frost in combination with dry deposition may result in very strong localized acidities.     

Inhibitory effect of a quercetin metabolite, quercetin 3-O-beta-D-glucuronide, on lipid peroxidation in liposomal membranes.

To study the antioxidant activity of quercetin 3-O-beta-D-glucuronide (Q3GA), which is one of the quercetin metabolites in the blood after intake of quercetin-rich food, the inhibitory effect of Q3GA on lipid peroxidation was estimated using phosphatidylcholine large unilamellar vesicles (PC LUV) as a biomembrane model. Iron ion, an aqueous peroxyl radical generator, a peroxynitrite generator, or lipoxygenase was used as the inducer of lipid peroxidation. In all cases, Q3GA inhibited lipid peroxidation significantly, although its inhibitory effect was lower than that of quercetin aglycon. The ultrafiltration of PC LUV containing Q3GA revealed that Q3GA has low but significant affinity with the membranes of phospholipid bilayers. It is therefore likely that Q3GA acts as an efficient antioxidant in membranous lipid peroxidation through its localization in the phospholipid bilayer. This conjugated quercetin metabolite seems to retain the ability to protect cellular and subcellular membranes from peroxidative attack by reactive oxygen species and peroxidative enzymes.     

Moscatilin from Dendrobium nobile, a naturally occurring bibenzyl compound with potential antimutagenic activity.

A bibenzyl compound that possesses antimutagenic activity was isolated from the storage stem of Dendrobium nobile. The isolated compound suppressed the expression of the umu gene following the induction of SOS response in Salmonella typhimurium TA1535/pSK1002 that have been treated with various mutagens. The suppressive compound was mainly localized in the n-hexane extract fraction of the processed D. nobile. This n-hexane fraction was further fractionated by silica gel column chromatography, which resulted in the purification and subsequent identification of the suppressive compound. EI-MS and (1)H and (13)C NMR spectroscopy were then used to delineate the structure of the compound that confers the observed antimutagenic activity. Comparison of the obtained spectrum with that found in the literature indicated that moscatilin is the secondary suppressive compound. When using 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide) as the mutagen, moscatilin suppressed 85% of the umu gene expression compared to the controls at <0.73 micromol/mL, with an ID(50) value of 0.41 micromol/mL. Additionally, moscatilin was tested for its ability to suppress the mutagenic activity of other well-known mutagens such as 4-nitroquinoline-1-oxide (4NQO), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), UV irradiation, 3-amino-1,4-dimethyl-5H-pyrido[4,3b]indole (Trp-P-1), benzo[a]pyrene (B[a]P), and aflatoxin B(1) (AFB(1)). With all of the aforementioned chemicals or treatments, moscatilin showed a dramatic reduction in their mutagenic potential. Interestingly, moscatilin almost completely suppressed (97%) the AFB(1)-induced SOS response at concentrations <0.73 micromol/mL, with an ID(50) of 0.08 micromol/mL. Finally, the antimutagenic activities of moscatilin against furylfuramide and Trp-P-1 were assayed by the Ames test using the S. typhimurium TA100 strain. The results those experiments indicated that moscatilin demonstrated a dramatic suppression of the mutagenicity of only Trp-P-1 but not furylfuramide.