In vitro inhibition of human cGMP-specific phosphodiesterase-5 by polyphenols from red grapes

A moderate consumption of red wine may reduce the risk of cardiovascular diseases via wine-derived phenolic compounds. A variety of biological mechanisms have been proposed for wine-derived phenolic compounds including nitric oxide-mediated vasorelaxation. This study examined whether the vasodilating effect of wine-derived phenolic compounds was associated with the inhibition of phosphodiesterases (PDEs) and, in particular, PDE5. For this purpose, human recombinant PDE5A1 isoform was prepared by expression of the full-length cDNA of PDE5A1 into COS-7 cells. Red wine and the extracts from grape skin inhibited PDE5A1 activity, whereas the seed extracts had a negligible effect. The mixture of anthocyanins inhibited the enzyme activity (IC50 = 11.6 microM), with malvidin-3-O-beta-glucoside (IC50 = 35.4 microM) and malvidin (IC50 = 24.9 microM) the most potent among the monoglucosides and aglycons, respectively. trans-Resveratrol and trans-piceid exhibited negligible effects, whereas hydroxycinnamates were completely inactive. These results indicate that polyphenols-induced vasorelaxation may also be sustained by smooth muscle PDE inhibition by anthocyanins present in red wines and grapes.     

2-fluoroabscisic acid analogues: their synthesis and biological activities

Fluorine was introduced into the 2-position of the side chain of abscisic acid (ABA) analogues by Wittig reaction of alpha-ionone derivatives with ethyl triethylphosphono-2-fluoroacetate. The effects of the fluorinated analogues were evaluated on inhibition of cress seed germination and inhibition of gibberellin-inducible alpha-amylase induction in embryoless barley half-seeds. (2E, 4E)-2-Fluoro-5-(1'-hydroxy-2',6', 6'-trimethyl-2'-cyclohexen-1'-yl)-3-methyl-2,4-pentadienoic acid (5b) showed potent inhibitory activity at the same level as ABA in the cress seed germination test, and 5b also inhibited gibberellin-inducible alpha-amylase induction at 4 x 10(-)(6), 3 times the concentration of ABA (1 x 10(-)(6)) for 50% inhibition of alpha-amylase production. 5b also showed dehydrin induction activity. These results indicate that fluorinated ABA analogues mimic ABA action and can be a lead for a plant growth regulator which regulates plant growth or protects plants from environmental stresses.     

Preparation of molecularly imprinted polymers using anacardic acid monomers derived from cashew nut shell liquid

The objective of this work was to use monomers from cashew ( Anacardium occidentale L.) nut shells to develop molecularly imprinted polymers. Cashew nut shell liquid (CNSL) is a cheap and renewable agro byproduct consisting of versatile monomers. Solvent-extracted CNSL contains over 80% anacardic acid (AnAc) with more than 90% degree of unsaturation in its C 15 side chain. From AnAc monomer, anacardanyl acrylate (AnAcr) and anacardanyl methacrylate (AnMcr) monomers were synthesized and their chemical structures were characterized by Fourier transform IR and NMR. Different imprinted bulk polymers based on AnAc, AnAcr, and AnMcr functional monomers have been prepared. In the present study, each functional monomer was separately copolymerized in toluene with ethylene glycol dimethacrylate and divinylbenzene as cross-linkers, using racemic propranolol as a model template. While the AnAc based polymer revealed a meager rebinding ability, the imprinted polymers made from AnAcr and AnMcr displayed highly specific propranolol binding. At a polymer concentration of 2 mg/mL, AnAcr and AnMcr based imprinted polymers were able to bind over 50% of trace propranolol (initial concentration 1.2 nM). Under the same condition propranolol uptake by the two nonimprinted control polymers was less than 20%. Chiral recognition properties of these polymers were further confirmed using tritium-labeled (S)-propranolol as a tracer in displacement experiments, suggesting that the apparent affinity of the imprinted chiral sites for the correct enantiomer is at least 10 times that of the mismatched (R)-propranolol. Moreover, cross reactivity studies of these polymers showed that the (S)-imprinted sites have higher cross-reactivity toward (R, S)-metoprolol than (R)-propranolol and (R)-timolol.     

Isolation of anacardic acid from natural cashew nut shell liquid (CNSL) using supercritical carbon dioxide

Solvent extracted cashew nut shell liquid (CNSL), conventionally known as natural CNSL, is a mixture of several alkenyl phenols. One of these alkenyl phenols is anacardic acid, which is present at the highest concentration. In view of anticipated industrial applications of anacardic acid, the objective of this work was to isolate anacardic acid from natural CNSL by supercritical carbon dioxide (scCO 2). In this study, the solubility data for natural CNSL in scCO 2 under a range of operating conditions of pressure (100, 200, and 300 bar), temperature (40 and 50 degrees C), and CO 2 flow rate (5, 10, and 15 g min (-1)) were established. The best scCO 2 working conditions were found to be 50 degrees C and 300 bar at a flow rate of 5 g min (-1) CO 2. Using 3 g of sample (CNSL/solid adsorbent = 1/2) under these scCO 2 conditions, it was possible to quantitatively isolate high purity anacardic acid from crude natural CNSL (82% of total anacardic acid) within 150 min. The anacardic acid isolated by scCO 2 was analyzed by different spectroscopic techniques (UV-vis, FT-IR, and (1)H NMR) and HPLC analysis, indicating that the anacardic acid isolated by scCO 2 has better quality than that obtained through a conventional method involving several chemical conversion steps.     

Antifeedant activity of some pentacyclic triterpene acids and their fatty acid ester analogues

The 3-O-fatty acid ester derivatives (C(12)-C(18)) of two pentacyclic triterpenic acids, ursolic acid and oleanolic acid, were synthesized under mild esterification conditions in excellent yields (80-85%) and screened for their antifeedant activity, together with the parent acids, against the agricultural pest tobacco caterpillar larvae (Spodoptera litura F) in a no-choice laboratory study. The Urs-12-ene-28-carboxy-3beta-octadecanoate and olean-12-ene-28-carboxy-3beta-hexadecanoate were found to exhibit exceptionally potent antifeedant activities at 50 microg/cm(2) concentration, even after 48 h.     

Bioactivity of anacardic acid against colorado potato beetle (Leptinotarsa decemlineata) larvae

Anacardic acid (2-hydroxy-6-alkylbenzoic acid) produced and secreted from glandular trichomes of zonal geranium (Pelargonium x hortorum; Geraniaceae family) provides resistance to small pests (aphids and spider mites). To assess the potential bioactivity of anacardic acid against larger insect pests and to determine if an alternate mode of application (ingestion rather than the topical application) could impart resistance to pests, the effects of anacardic acid consumption on the development of Colorado potato beetle larvae were tested. Analysis of dose-response curves indicated a significant effect on weight gain and mortality. Assessment of food preference (treated versus untreated) indicated larvae avoid food containing anacardic acid and have a lower feeding rate on food containing anacardic acid. On the basis of these results, it is suggested that anacardic acid, applied as a chemical spray or through bioengineering production in crop plants, may provide a new tool in the arsenal to minimize damage to plants caused by pests.     

Synthesis of photosynthesis-inhibiting nostoclide analogues

A series of 34 3-benzyl-5-(arylmethylene)furan-2(5H)-ones, designed using the naturally occurring toxins nostoclides as a lead structure, was synthesized as potential inhibitors of the photosynthetic electron transport. All compounds were fully characterized by IR, NMR (1H and 13C), and MS spectrometry. HMBC and HSQC bidimensional experiments allowed 13C and 1H assignments. Their biological activities were evaluated in vitro as the ability to interfere with light-driven reduction of ferricyanide by isolated spinach chloroplasts. About two-thirds of the compounds exhibited inhibitory properties in the micromolar range against the basal electron flow from water to K3[Fe(CN)6]. The inhibitory potential of these 3-benzyl-5-(arylmethylene)furan-2(5H)-one lactones is higher than that of other nostoclide analogues previously synthesized in the same laboratories.     

Synthesis and antifeeding activities of Tonghaosu analogues

Tonghaosu (1), a lead for a botanical antifeedant, and its 22 analogues were synthesized according to a previously reported concise and straightforward procedure. The structures of all new compounds were confirmed by NMR, IR, MS, and HREIMS or elemental analysis. Their insect antifeedant activities against the large white butterfly (Pieris brassicae L.) were examined, and six analogues (Z- and E-6h and Z-isomers of 6i-l), which contain 1,3-diyn or 3,4-methylenedioxyphenyl acetylene group, showed considerable antifeedant activity. Interestingly, Z-isomers of 6i-k are much more active than their corresponding E-isomers.     

Synthesis and antifungal activity of novel sclerotiorin analogues

Sclerotiorin 1, first isolated from Penicillium sclerotiorum, has weak antifungal activity and belongs to the azaphilone-type family of natural products. Several series of sclerotiorin analogues were designed and synthesized with the aim of discovering novel fungicides with improved activity. The syntheses involved two key steps, cycloisomerization and then oxidation, and used a simple and efficient Sonogashira cross-coupling reaction to construct the required functionalized precursor. With sclerotiorin as a control, the activities of the newly synthesized analogues were evaluated against seven fungal pathogens, and several promising candidates (compounds 3a?, 3d?, 3e?, 3f? and 3k?) with greater activity and simpler structures than sclerotiorin were discovered. In addition, preliminary structure-activity relationships were studied, which revealed that not only the chlorine or bromine substituent at the 5-position of the nucleus but also the phenyl group at the 3-position and the substituent pattern on it contributed crucially to the observed antifungal activity. Analogues with a methyl substituent at the 1-position have reduced levels of activity, while those with a free hydroxyl group in place of acetoxy at the quaternary center of the bicyclic ring system retain activity.     

Synthesis and antifungal action of new tricyclazole analogues

Melanins are very important pigments for the survival and longevity of fungi, so their biosynthesis inhibition is a new biochemical target aiming at the discovery of selective fungicides. In this work is described the synthesis of new pyrazolo-thiazolo-triazole compounds, analogues of tricyclazole (a commercial antifungal product that acts by inhibiting melanin synthesis), and their biological activity was studied on some dermatophytes and phytopathogens. The compounds poorly inhibited the growth and pigmentation of fungi tested and were less efficient than tricyclazole. Electron microscopy on Botrytis cinerea showed that treatment with the most active compound caused abnormally thickened and stratified walls in fungi, whose ultrastructure was, in contrast, generally normal. The fungus treated with tricyclazole, on the other hand, appeared to be drastically altered, so as to become completely disorganized. These results suggest that the new azole compounds employ an action mechanism similar to that of other azoles, but dissimilar to that of tricyclazole.     

Synthesis and fungicidal evaluation of novel chalcone-based strobilurin analogues

Strobilurin derivatives have become one of the most important classes of agricultural fungicide due to a novel action mode, wide fungicidal spectrum, lower toxicity toward mammalian cells, and environmentally benign characteristics. To discover new strobilurin analogues with high activity against resistant pathogens, a series of new chalcone-based strobilurin derivatives are designed and synthesized by integrating a chalcone scaffold with a strobilurin pharmacophore. The preliminary bioassay showed that some of the chalcone analogues exhibited good in vivo fungicidal activities against Pseudoperoniospora cubensis and Sphaerotheca fuliginea at the dosage of 200 microg mL(-1). Two compounds, (E)-methyl 2-[2-({3-[(E)-3-(2-chlorophenyl)acryloyl]phenoxy}methyl)phenyl]-3-methoxyacrylate (1e) and (E)-methyl 2-[2-({3-[(E)-3-(3-bromophenyl)acryloyl]phenoxy}methyl)phenyl]-3-methoxyacrylate (1l), were found to display higher fungicidal activities against P. cubensis (EC90=118.52 microg mL(-1) for 1e and EC90=113.64 microg mL(-1) for 1l) than Kresoxim-methyl (EC90=154.92 microg mL(-1)) and were identified as the most promising candidates for further study. The present work demonstrated that strobilurin analogues containing chalcone as a side chain could be used as a lead structure for further developing novel fungicides. To our knowledge, this is the first report about the syntheses and fungicidal activities of chalcone-based strobilurin derivatives.     

Hydrogen and aluminum inhibition of soybean root extension from limed soil into acid subsurface solutions

Soybean [Glycine max (L.) Merr. cv. ‘Ransom'] root elongation under varying concentrations of solution hydrogen (H) and aluminum (Al) was investigated in a vertical split‐root system. Roots extending from a limed and fertilized soil compartment grew for 12 days into a subsurface compartment with solutions adjusted to either different pH values from 3.7 to 5.5 or a factorial combination of pH (4.0,4.6, and 5.2) and Al (0,7.5, 15, and 30 μM) levels. Ionic forms of Al were estimated with GEOCHEM and solution Al was determined with ferron. Boron (B) (18.5 μM) and zinc (Zn) (0.5 μM) were supplied to all solution treatments, in addition to 2000 μM Ca, after preliminary studies at pH 5.2 without Al indicated that their omission inhibited length of tap roots and their laterals in the subsurface compartment. Both H⁺ and Al inhibited the length of lateral roots more than tap roots. Lateral roots failed to develop on tap roots at pH<4.3 or in treatments with 30 μM Al. Relative tap root length (RRL) among treatments receiving Al correlated with Al as measured by reaction with ferron for 30s. Ferron‐reactive Al was correlated to GEOCHEM‐predicted Al³⁺ activity (r=0.99). A 50% reduction in RRL occurred with either 2.1 μM Al³⁺ activity or 4.9 uM ferron‐reactive Al. The absence of shoot and soil‐root biomass differences among solution treatments in the split‐root system indicated that differences in root growth in the subsurface compartment were not directly confounded with differences in top growth.     

Avocadofurans and their tetrahydrofuran analogues: comparison of growth inhibitory and insecticidal activity

The importance of the double bonds in the furan ring of avocadofurans with relation to their insecticidal activity was examined. The insecticidal activity of two naturally occurring avocadofurans, 2-(pentadecyl)furan and 2-(heptadecyl)furan, was compared to the toxicity of five tetrahydrofurans with alkyl chains at position 2 and varying side chains from 14 to 18 carbons. We found that eliminating the sites of unsaturation in the furan ring of avocadofurans significantly reduced the detrimental effects on the mortality and growth of the generalist insect herbivore Spodoptera exigua. In 7-day bioassays, S. exigua larvae were significantly more affected when fed a diet containing avocadofurans as compared to a larvae fed diet treated with the analogous tetrahydrofurans. Although larvae fed with the tetrahydrofurans showed reduced growth as compared to controls, larval mortality was not significantly increased. We conclude that the double bonds in the furan ring of avocadofurans play an important role in their insecticidal effects.     

条斑紫菜多糖超声提取及其对U937细胞生长抑制影响的研究

采用超声辅助提取技术进行条斑紫菜多糖的提取,并进行柱层析纯化。经正交试验确定的条斑紫菜粗多糖提取的最佳工艺条件为：15 g干紫菜粉加入2 L水,提取液循环转速为12 r/s,提取液温度为40℃,超声功率为800 W,超声处理时间/超声间隙为3 s/2 s和超声全程时间为2 h,对应的得率为26.29%。粗多糖分别经50%、60%、70%和80%乙醇分步醇沉,沉淀出四种不同级分的半纯品,用DEAE-52纤维素和Sephadex G-200将其中两种多糖半纯品纯化,分别得到5个和3个不同分子量的级分,分别考察了这两种紫菜多糖半纯品及各自分级产物对人血癌细胞U937细胞的体外生长抑制的抗肿瘤试验,结果前者及其分级产物对U937细胞生长有一定的抑制作用,后者及其分级产物则有一定的促进作用。     

5-caffeoylquinic acid and caffeic acid down-regulate the oxidative stress- and TNF-alpha-induced secretion of interleukin-8 from Caco-2 cells

Although chlorogenic acid (CHA) easily reaches a millimolar level in the gastrointestinal tract because of its high concentration in coffee and fruits, its effects on intestinal epithelial cells have been little reported. We investigated in this study the down-regulative effects of 5-caffeoylquinic acid (CQA), the predominant isomer of CHA, on the H(2)O(2-) or TNF-alpha-induced secretion of interleukin (IL)-8, a central pro-inflammatory chemokine involved in the pathogenesis of inflammatory bowel diseases, in human intestinal epithelial Caco-2 cells. After the cells had been pre- and simultaneously treated with CQA, the oversecretion of IL-8 and overexpression of its mRNA induced by H(2)O(2) were significantly suppressed in a dose-dependent manner in the range of 0.25-2.00 mmol/L. We further found that a metabolite of CQA, caffeic acid (CA), but not quinic acid, significantly inhibited the H(2)O(2)-induced IL-8 secretion and its mRNA expression in the same dose-dependent manner. Both CQA and CA suppressed the TNF-alpha-induced IL-8 secretion as well. Caffeic acid at 2.00 mmol/l was able to absolutely block the H(2)O(2)- or TNF-alpha-induced oversecretion of IL-8 in Caco-2 cells. However, CQA and CA did not suppress the TNF-alpha-induced increase in the IL-8 mRNA expression, indicating that the suppressive mechanisms are different between TNF-alpha-induced and H(2)O(2)-induced IL-8 production models. These results suggest that the habit of drinking coffee and/or eating fruits with a high CHA content may be beneficial to humans in preventing the genesis of inflammatory bowel diseases.     

Calcium alleviation of hydrogen and aluminum inhibition of soybean root extension from limed soil into acid subsurface solutions

Alleviation by calcium (Ca) of inhibition of soybean [Glycine max (L.) Merr. cv. ‘Ransom'] root elongation by hydrogen (H) and aluminum (Al) was evaluated in a vertical split‐root system. Roots extending from a limed and fertilized soil compartment grew for 12 days into a subsurface compartment containing nutrient solution with treatments consisting of factorial combinations of either pH (4.0, 4.6, and 5.5) and Ca (0.2, 2.0, 10, and 20 mM), Al (7.5, 15, and 30 μM) and Ca (2.0,10, and 20 mM) at pH 4.6, or Ca (2, 7, and 12 mM) levels and counter ions (SO₄ and Cl) at pH 4.6 and 15 μM Al. Length of tap roots and their laterals increased with solution Ca concentration and pH value, but decreased with increasing Al level. Length of both tap and lateral roots were greater when Ca was supplied as CaSO₄ than as CaCl₂, but increasing Ca concentration from 2 to 12 mM had a greater effect on alleviating Al toxicity than Ca source. In the absence of Al, relative root length (RRL) of tap and lateral roots among pH and Ca treatments was related to the Ca:H molar activity ratio of solutions (R²≥0.82). Tap and lateral RRL among solutions with variable concentrations of Al and Ca at pH 4.6 were related to both the sum of the predicted activities of monomeric Al (R²≥0.92) and a log‐transformed and valence‐weighted balance between activities of Ca and selected monomeric Al species (R²≥0.95). In solutions with 15 μM Al at pH 4.6, response of tap and lateral RRL to variable concentrations of CaSO₄ and CaCl₂ were related to predicted molar activity ratios of both Ca:Al³⁺ (R²≥0.89) and Ca:3 monomeric Al (R²≥0.90), provided that AISO₄ and AI(SO₄)₂ species were excluded from the latter index. In all experiments H and Al inhibited length of lateral roots more than tap roots, and a greater Ca:H or Ca:Al concentration ratio was required in solutions to achieve similar RRL values as tap roots.     

Identification of 4-O-5'-coupled diferulic acid from insoluble cereal fiber

The extracts of saponified cereal fibers of whole grains of corn (Zea mays cv. microsperma KOERN.), wheat (Triticum aestivum L.), spelt (Triticum spelta L.), and rice (Oryza sativa L.) were investigated for dehydrodimers of ferulic acid using gas-liquid chromatography (GLC) with mass spectrometric detection (GLC-MS) and flame ionization detection (GLC-FID). In addition to the 8,5'-, 8, 8'-, 5,5'-, and 8-O-4'-coupled diferulic acids previously identified from other plant materials the 4-O-5'-coupled diferulic acid (E)-3-[4-[(E)-2-carboxyvinyl]-2-methoxyphenoxy]-4-hydroxy-5-methoxyci nnamic acid (4-O-5'-DFA) was identified in all fibers investigated. This new diferulate was authenticated by comparison of its mass spectrum and its relative GLC retention time with those of the synthesized compound. Semiquantitative determination of 4-O-5'-DFA showed that it is present at 8-30 microg/g, approximately 70-100 times lower concentrations than the sum of 8,5'-coupled diferulic acids, the major diferulic acids in the investigated fibers.     

Studies on umami taste. Synthesis of new guanosine 5'-phosphate derivatives and their synergistic effect with monosodium glutamate

A number of N2-alkyl and N2-acyl derivatives of guanosine 5'-phosphate (GMP) have been synthesized and tested for their synergistic effect with monosodium L-glutamate (MSG), the prototypical substance imparting umami taste to savory-based foods. Capacities to enhance the taste intensity of MSG (gamma values) were estimated through subjective comparisons of MSG/nucleotide mixtures in water with appropriate solutions of MSG alone. Assuming beta = gamma[nucleotide]/gamma[IMP], beta values of the N2-substituted GMPs were found in the range 1.2-5.7. Such values appear to be related to the chain length of the substituent in the 2-position of the purine nucleus and dependent on the replacement of a CH 2 group with an S atom and/or with an alpha-CO group. These findings indicate that the exocyclic NHR group of the guanine moiety is actively implicated in the synergism between GMP derivatives and MSG. Theoretical calculations suggest that an anti conformation is probably assumed by ribonucleotide molecules interacting with umami receptors.     

Kinetic study of oxalic acid inhibition on enzymatic browning

Oxalic acid has a strong antibrowning activity. The inhibitory pattern on catechol-PPO model system appeared to be competitive, with a K(i) value of 2.0 mM. When the PPO was incubated with oxalic acid, the activity was not recovered via dialysis, but the inactivated enzyme partially recovered its activity when cupric ion was added. Comparing the relative antibrowning effectiveness of oxalic acid with other common antibrowning agents, oxalic acid with I(50) value of 1.1 mM is as effective as kojic acid and more potent than cysteine and glutathione.