Synthesis and herbicidal activity of diphenyl ether derivatives containing unsaturated carboxylates

A series of novel diphenyl ether derivatives containing unsaturated carboxylates were designed and synthesized. Their structures were identified by (1)H nuclear magnetic resonance and elemental analyses. The bioassays indicated that the compounds 5b and 5c exhibited good herbicidal activities against velvetleaf at a concentration of 30-40 g/hm(2). The relationship between structure and herbicidal activity was also discussed. Among unsaturated carboxylates group, butenoate is the most promising one. Amonst them, 4-ethoxy-4-oxobutenyl 5-(2-chloro-4-(trifluoromethyl)phenoxy)-2-nitrobenzoate 5b was identified as the most promising candidate due to its high protoporphyrinogen IX oxidase inhibition effect (pI(50) = 6.64) and good herbicidal activity against broadleaf weeds with selectivity to soybean and low toxicity to mammals.     

Inhibition of key digestive enzymes by cocoa extracts and procyanidins

This study determined the in vitro inhibitory effects of cocoa extracts and procyanidins against pancreatic α-amylase (PA), pancreatic lipase (PL), and secreted phospholipase A(2) (PLA(2)) and characterized the kinetics of such inhibition. Lavado, regular, and Dutch-processed cocoa extracts as well as cocoa procyanidins (degree of polymerization (DP) = 2-10) were examined. Cocoa extracts and procyanidins dose-dependently inhibited PA, PL, and PLA(2). Lavado cocoa extract was the most potent inhibitor (IC(50) = 8.5-47 μg/mL). An inverse correlation between log IC(50) and DP (R(2) > 0.93) was observed. Kinetic analysis suggested that regular cocoa extract, the pentamer, and decamer inhibited PL activity in a mixed mode. The pentamer and decamer noncompetitively inhibited PLA(2) activity, whereas regular cocoa extract inhibited PLA(2) competitively. This study demonstrates that cocoa polyphenols can inhibit digestive enzymes in vitro and may, in conjunction with a low-calorie diet, play a role in body weight management.     

Antioxidant activity of extracts of black sesame seed (Sesamum indicum L.) by supercritical carbon dioxide extraction

Antioxidant activities of extracts derived from sesame seed by supercritical carbon dioxide (SC-CO(2)) extraction and by n-hexane were determined using alpha,alpha-diphenyl-beta-picylhydrazyl (DPPH) radical scavenging and linoleic acid system methods. The highest extracted yield was given at 35 degrees C, 40 MPa, and a CO(2) flow rate of 2.5 mL min(-1) by an orthogonal experiment. The yields of extracts increased with increasing pressure, and yields at 40 and 30 MPa were higher than that by solvent extraction at 46.50%. Results from the linoleic acid system showed that the antioxidant activity follows the order: extract at 35 degrees C, 20 MPa > BHT > extract at 55 degrees C, 40 MPa > extract at 55 degrees C, 30 MPa > Trolox > solvent extraction > alpha-tocopherol. The SC-CO(2) extracts exhibited significantly higher antioxidant activities comparable to that by n-hexane extraction. The extracts at 30 MPa presented the highest antioxidant activities assessed in the DPPH method. At 20 MPa, the EC(50) increased with temperature, which indicated that the antioxidant activity was decreased in a temperature-dependent manner. The significant differences of antioxidant activities were found between the extracts by SC-CO(2) extraction and n-hexane. However, no significant differences were exhibited among the extracts by SC-CO(2) extraction. The vitamin E concentrations were also significantly higher in SC-CO(2) extracts than in n-hexane extracts, and its concentrations in extracts corresponded with the antioxidant activity of extracts.     

Inhibition by soil humic acids of native and acetylated proteolytic enzymes

The activities of acetyltrypsin and acetylcarboxypeptidase were unaffected by neutralized solutions of soil humic acids in concentrations which markedly inhibited the non-acetylated enzymes. Further, acetyltrypsin, unlike trypsin, did not coprecipitate with humic acids. The results support the conclusion that humic acids bind the proteases by a cationexchange mechanism whereby protease amino groups are linked to humic acid carboxyl groups.     

复合氨基酸肥料增效剂对NaCl胁迫下小白菜种子萌发和苗期生长的影响

【目的】作物种子萌发期和苗期是对盐胁迫最为敏感的时期,盐分过高会严重影响作物种子萌发和幼苗生长。本研究以谷氨酸尾液为主要材料开发了复合氨基酸肥料增效剂(简称增效剂),并研究其在盐(NaCl)胁迫条件下对种子萌发、苗期生长和生理指标的影响,旨在为谷氨酸尾液在盐碱土地区的推广应用提供科学依据和理论指导。【方法】以小白菜种子和幼苗为供试材料,分别进行种子萌发试验和水培试验。1)种子萌发试验:采用标准发芽试验法,种子分别经0、0.05、0.1、0.2、0.4、0.8 g/L增效剂浸种后,分别移至含0、25、50、75mmol/L NaCl溶液中萌发,测定发芽势、发芽率、胚根长和胚芽长。2)苗期水培试验:选取整齐一致的幼苗,缓苗后同时加入与萌发试验浓度一致的增效剂和NaCl溶液,在盐害明显后取样测定鲜重、SPAD值、根长、株高以及叶片过氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性、过氧化物酶(POD)活性、丙二醛(MDA)含量、超氧阴离子自由基(O_2~-)产生速率、脯氨酸(Pro)含量等盐胁迫评价指标。【结果】在0~75 mmol/L NaCl范围内,NaCl浓度越高对小白菜种子萌发和幼苗生长的抑制作用越强,一定浓度的增效剂可不同程度地缓解NaCl对种子萌发和幼苗的胁迫。1)在无盐胁迫下,低浓度增效剂对种子萌发具有轻微的抑制作用,而高浓度增效剂则会显著抑制种子萌发;在同一浓度NaCl胁迫下,随增效剂浓度的增加,小白菜种子发芽势、发芽率、胚根长和胚芽长均表现出先上升后下降的变化规律,增效剂浓度为0.2 g/L时效果最佳,而在0.4 g/L和0.8 g/L时则会抑制小白菜种子萌发。2)在无盐胁迫下,随增效剂浓度增加对小白菜生长表现出先促进后抑制的效果,以0.1 g/L用量效果最好;在同一浓度NaCl胁迫下,增效剂浓度为0.05 g/L时,提高了苗期小白菜鲜重、SPAD值,并促进了根伸长和茎伸展,同时提高叶片SOD、POD、CAT活性和Pro含量,并降低MDA含量和产生速率;之后随增效剂浓度的增加小白菜幼苗鲜重、SPAD值、根长和株高均表现出持续下降的趋势,而SOD、POD、CAT活性,Pro含量表现出先持平后下降的变化规律,产生速率和MDA含量则表现出先上升后平稳的趋势,增效剂浓度达到0.4 g/L和0.8 g/L时小白菜幼苗生长受到明显抑制。【结论】在无盐胁迫条件下,低浓度(≤0.2 g/L)复合氨基酸肥料增效剂可轻微抑制小白菜种子萌发,但在25~75 mmol/L NaCl胁迫条件下,则可明显促进种子萌发、提高种子发芽质量;而在相同盐胁迫条件下,低浓度复合氨基酸肥料增效剂可明显促进小白菜幼苗生长,提高叶片抗氧化酶活性、维持渗透调节物质Pro含量、增强光合作用等,以0.05 g/L效果最佳。     

Determination of organic acids and phosphate in soil aqueous extracts by capillary zone electrophoresis

Abstract

Organic acids and phosphate were determined in soil aqueous extracts by capillary zone electrophoresis using indirect UV detection. The electrolyte system was 10 mM sodium benzoate with 0.5 mM tetradecyltrimetyl‐ammonium bromide as flow modifier at pH 4.0, pH 4.5, or pH 5.0. This methodology was adequate to determine organic acids and phosphate in soil samples, but the major inorganic anions interfered in the determination. In all the samples of soil extracts, the presence of phosphate was detected. Acetate was found in most of the samples and lactate and formate in some of them.     

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.     

Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models.

Antioxidant-rich fractions were extracted from pomegranate (Punica granatum) peels and seeds using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants using various in vitro models, such as beta-carotene-linoleate and 1,1-diphenyl-2-picryl hydrazyl (DPPH) model systems. The methanol extract of peels showed 83 and 81% antioxidant activity at 50 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. Similarly, the methanol extract of seeds showed 22.6 and 23.2% antioxidant activity at 100 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. As the methanol extract of pomegranate peel showed the highest antioxidant activity among all of the extracts, it was selected for testing of its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 56, 58, and 93.7% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 100 ppm. This is the first report on the antioxidant properties of the extracts from pomegranate peel and seeds. Owing to this property, the studies can be further extended to exploit them for their possible application for the preservation of food products as well as their use as health supplements and neutraceuticals.     

Antioxidant activity of extracts, condensed tannin fractions, and pure flavonoids from Phaseolus vulgaris L. seed coat color genotypes

Twelve different seed coat color genotypes of Phaseolus vulgaris L. were extracted and pure flavonoids isolated from 10 of these. The seed coat methanol extracts, tannin fractions, and pure flavonoids all displayed antioxidant activity in a fluorescence-based liposome assay. The relatively high activity of the condensed tannin (proanthocyanidin) fractions indicates that these may play an important role in the overall activity of the extracts. This activity also indicates that although these polyphenols cause problems in digestibility, they may be important dietary supplements with beneficial health effects. The pure anthocyanins delphinidin 3-O-glucoside (1), petunidin 3-O-glucoside (2), and malvidin 3-O-glucoside (3) and the flavonol quercetin 3-O-glucoside (4) isolated from seed coats also had significantly higher antioxidant activity than the Fe(2+) control. The activity of kaempferol 3-O-glucoside (5) was not different from that of the Fe(2+) control. These findings suggest that variously colored dry beans may be an important source of dietary antioxidants.     

Inhibition of reactive nitrogen species effects in vitro and in vivo by isoflavones and soy-based food extracts

Recent studies have shown that soy isoflavone inhibits inducible nitric oxide (NO) synthase activities and is reported to have peroxynitrite scavenging ability. Consequently, we investigated whether isoflavones (daidzein and genistein) and extracts from soy-based products (miso, soymilk, tofu, soy sprout, black soybean, soybean, and yuba) would inhibit the reactive nitrogen species (RNS) effect in vitro and in vivo. In the in vitro experiments [including the protection of cellular DNA from peroxynitrite or sodium nitroprusside damage, an inhibitory effect on nitric oxide production from lipopolysaccharide (LPS)-induced RAW 264.7 cells, and nitric oxide scavenging ability], extracts from soy-based foods showed a potent antioxidant activity and an inhibiting effect on RNS activity. These effects were correlated with total isoflavone content. In the in vivo experiments, rats were given isoflavones (4.0 mg/kg bw) or soy-based product extracts (1.0 g/kg bw) orally for 1 week and were injected with vehicle H(2)O (1 mL/kg bw) or LPS (10 mg/kg bw) on the day 7. Twelve hours after treatment, the rats were killed, and blood serum was collected for analysis. The intraperitoneal administration of LPS resulted in an increase in serum nitrite, nitrate, and nitrotyrosine concentrations. These are stable metabolite end products of nitric oxide, to 4-, 16-, and 5-fold levels, (4, 10 microM and 58 +/- 14 pmol/mL), of the placebo control, respectively. Results showed that oral administration of isoflavones and extracts from soy-based products significantly decreased serum nitrite, nitrate, and nitrotyrosine levels in LPS-induced rats. This study demonstrates that soy isoflavone supplementation may inhibit RNS-induced oxidation both in vitro and in vivo.     

Inhibition of cancer cell proliferation in vitro by fruit and berry extracts and correlations with antioxidant levels

The effects of 10 different extracts of fruits and berries on cell proliferation of colon cancer cells HT29 and breast cancer cells MCF-7 were investigated. The fruits and berries used were rosehips, blueberries, black currant, black chokeberries, apple, sea buckthorn, plum, lingonberries, cherries, and raspberries. The extracts decreased the proliferation of both colon cancer cells HT29 and breast cancer cells MCF-7, and the effect was concentration dependent. The inhibition effect for the highest concentration of the extracts varied 2-3-fold among the species, and it was in the ranges of 46-74% (average = 62%) for the HT29 cells and 24-68% (average = 52%) for the MCF-7 cells. There were great differences in the content of the analyzed antioxidants in the extracts. The level of the vitamin C content varied almost 100-fold, and the content of total carotenoids varied almost 150-fold among the species. Also in the composition and content of flavonols, hydroxycinnamic acids, anthocyanins, and phenolics were found great differences among the 10 species. The inhibition of cancer cell proliferation seen in these experiments correlated with levels of some carotenoids and with vitamin C levels, present at levels that can be found in human tissues. The same inhibition of cell proliferation could not be found by ascorbate standard alone. This correlation might indicate a synergistic effect of vitamin C and other substances. In MCF-7 cells, the anthocyanins may contribute to the inhibition of proliferation.     

Analysis of Piperaceae germplasm by HPLC and LCMS: a method for isolating and identifying unsaturated amides from Piper spp extracts

A method for extraction and high performance liquid chromatography-mass spectrometer (HPLC-MS) analysis of the medicinally important genus Piper (Piperaceae) was developed. This allows for a rapid and accurate measure of unsaturated amides, or piperamides, in black pepper, Piper nigrum L., and in wild species from Central America. Reflux extraction provided the highest recovery of piperine (>80%) from leaf and peppercorn material. HPLC analysis using a binary gradient of acetonitrile and water separated the major amide peaks between 5 and 12 min. Atmospheric pressure chemical ionization (APCI)-MS improved the detection limit to 0.2 ng, 10-fold below the 2 ng limit of the HPLC-diode array detector (DAD) based on linear standard curves between 0.1 and 250 microg/mL (R2 = 0.999). The HPLC-MS method identified pellitorine, piperylin, 4,5-dihydropiperlonguminine, piperlonguminine, 4,5-dihydropiperine, piperine, and pipercide. The biological activity of six Costa Rican Piper species assessed by mosquito larval bioassays correlated well with piperamide content.     

Rapid and sensitive quantification of amino acids in soil extracts by capillary electrophoresis with laser-induced fluorescence

A growing body of research is arguing that amino acids are key components of the soil nitrogen cycle. For example, we now know that many plants can take up intact amino acids, even in competition with soil microbes. Our growing recognition of the importance of amino acids is not matched by knowledge of the amounts and type of amino acids in the soil, certainly not in comparison with our encyclopaedic knowledge of inorganic N. The primary reason that less is known about amino acids than inorganic N is that measuring the amounts of individual amino acids with conventional chromatographic techniques is slow and typically requires extensive sample clean-up if KCl extracts are analysed. The aim of this study was to develop capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) as a more rapid alternative for measuring individual amino acids in KCl extracts of soil. The CE-LIF method separated 17 common amino acids within 12 min, with detection limits between 7 and 250 nM. One molar KCl extracts could be analysed without any sample clean-up or de-salting, and spike and recovery tests indicated that the complex matrix of soil extracts did not affect quantification. Further evidence of the suitability and robustness of the method came from the repeated analysis (n=5) of the same soil KCl extract. The relative standard deviation of migration times for replicate analyses were <0.2% while relative standard deviations for peak areas were <5%. To demonstrate application of the CE-LIF method to real world problems it was used to analyse amino acids in 1 M KCl extracts from a sub-alpine grassland and a Eucalyptus regnans forest. The most abundant amino acids were Ala, Gly and Arg. Other amino acids present at smaller concentrations or in a minority of samples were Asn, Cit, GABA, Glu, His, Phe, Leu, and Lys. The proposed CE-LIF method offers significant advantages over chromatographic methods via its rapidity, reproducibility and, most importantly, its ability to analyse crude KCl extracts.     

Inhibition of apple polyphenol oxidase activity by procyanidins and polyphenol oxidation products

The rate of consumption of dissolved oxygen by apple polyphenol oxidase in cider apple juices did not correlate with polyphenol oxidase activity in the fruits and decreased faster than could be explained by the decrease of its polyphenolic substrates. The kinetics parameters of a crude polyphenol oxidase extract, prepared from apple (Braeburn cultivar), were determined using caffeoylquinic acid as a substrate. Three apple procyanidin fractions of n 80, 10.5, and 4 were purified from the parenchyma of cider apples of various cultivars. Procyanidins, caffeoylquinic acid, (-)-epicatechin, and a mixture of caffeoylquinic acid and (-)-epicatechin were oxidized by reaction with caffeoylquinic acid o-quinone in order to form oxidation products. All the fractions were evaluated for their inhibitory effect on PPO activity. Native procyanidins inhibited polyphenol oxidase activity, the inhibition intensity increasing with n. The polyphenol oxidase activity decreased by 50% for 0.026 g/L of the fraction of n 80, 0.17 g/L of the fraction of n 10.5, and 1 g/L of the fraction of n 4. The inhibitory effect of oxidized procyanidins was twice that of native procyanidins. Oxidation products of caffeoylquinic acid and (-)-epicatechin also inhibited polyphenol oxidase.     

Evaluation of antioxidant activity and preventing DNA damage effect of pomegranate extracts by chemiluminescence method

The antioxidant activities of three parts (peel, juice, and seed) and extracts of three pomegranate varieties in China were investigated by using a chemiluminescence (CL) method in vitro. The scavenging ability of pomegranate extracts (PEs) on superoxide anion, hydroxide radical, and hydrogen peroxide was determined by the pyrogallol-luminol system, the CuSO4-Phen-Vc-H2O2 system, and the luminol-H2O2 system, respectively. DNA damage preventing the effect of PE was determined by the CuSO4-Phen-Vc-H2O2-DNA CL system. The results showed that the peel extract of red pomegranate had the best effect on the scavenging ability of superoxide anion because its IC50 value (4.01 +/- 0.09 microg/mL) was the lowest in all PEs. The seed extract of white pomegranate could scavenge hydroxide radical most effectively of the nine extracts (the IC50 value was 1.69 +/- 0.03 microg/mL). The peel extract of white pomegranate had the best scavenging ability on hydrogen peroxide, which had the lowest IC50 value (0.032 +/- 0.003 microg/mL) in the nine extracts. The seed extract of white pomegranate (the IC50 value was 3.67 +/- 0.03 microg/mL) was the most powerful on the DNA damage-preventing effect in all of the PEs. Also, the statistical analysis indicated that there were significant differences (at P < 0.05) among the extracts of the different varieties and parts in each system.     

Purification and identification of linoleic acid hydroperoxides generated by soybean seed lipoxygenases 2 and 3

It has been known that lipoxygenase (LOX) isozymes exhibit differences in product formation, but most product information to date is for LOX 1 among soybean (Glycine max) LOX isozymes. In this study, LOXs 2 and 3 were purified and used to generate hydroperoxide (HPOD) products in an in vitro system using linoleic acid as a substrate in the presence of either air or O2. The products were analyzed to determine their stereoisomeric characteristics. The control (no enzyme) showed only low levels of hydroperoxide production and no stereoisomeric specificity. LOX 2 shows high specificity in product formation, producing roughly 4 times more 13-HPOD than 9-HPOD, nearly all of which was 13-S(Z,E)-HPOD. LOX 3 produced more 9-HPOD than 13-HPOD at approximately a 2:1 ratio. No single stereoisomer was predominant among LOX 3 products. These results demonstrate that different isozymes of LOX have characteristic product profiles in in vitro reactions.     

Components of organic phosphorus in soil extracts that are hydrolysed by phytase and acid phosphatase

 Extracts were prepared from soil using water, 50 mM citric acid (pH ∼2.3) or 0.5 M NaHCO₃ (pH 8.5), and were incubated with excess phytase from Aspergillus niger to determine the amounts of labile P. Two A. niger phytase preparations were used: (1) a purified form which exhibited a narrow substrate specificity and high specific activity against phytate; and (2) a commercial preparation (Sigma) with activity against a broad range of P compounds. A comparatively large proportion (up to 79%, or 5.7 μg g^–1 soil) of the organic P (P_o) extracted with citric acid was hydrolysed by the commercial phytase, while between 28% and 40% (up to 3.1 μg g^–1 soil) was hydrolysed using purified phytase. By comparison, only small quantities of the P_o in water and NaHCO₃ soil extracts were enzyme labile. While extractable P_o was increased both with increasing concentrations of citric acid (up to 50 mM) and increasing pH (pH 2.3–6.0), enzyme-labile P increased only with citric acid concentration. The labile component of P_o in citric acid extracts from soils with contrasting fertiliser histories indicated that enzyme-labile P_o is a relatively large soil P pool and is potentially an important source of P for plants. Received: 29 October 1999     

Grain yield,seed weight,seed N concentration,and nodule activity of soybean as influenced by defoliation and N fertilizer

It is unknown if nitrogen (N) fertilizer application will ameliorate the yield loss associated with severe defoliation of soybean [Glycine max (L.) Merr.] at the R5 stage of growth. The objective of this field study was to investigate the interaction of N fertilization rate and extent of defoliation on soybean yield, seed weight, seed N concentration, and nodule activity. Field experiments were conducted in 1988 and 1989 on a Drummer silty clay loam (Typic Haplaquolls). Treatment variables were three cultivars: BSR 101, Chamberlain, and Elgin 87; three N fertilizer rates applied one day after defoliation: 0, 84, and 168 kg N ha^‐1 as urea; and three levels of defoliation: 0, 50, and 75%. Grain yield was not significantly affected by N rate but did decrease with defoliation. Fertilizer N did not ameliorate the yield reduction associated with defoliation. Seed weight decreased linearly with increasing defoliation. Plants exposed to the most severe defoliation produced seed which weighed 1 g 100^‐1 seed less than seed from nondefoliated plants. In 1989 seed weight of only the nondefoliated plants increased slightly with N rate, seed weight was not affected by N rate for any other year by defoliation treatment combination. Seed N concentration was not affected by N rate. Seed N concentration increased with defoliation in 1988 but not in 1989. Seed N concentration was not affected by defoliation in 1989. N fertilizer application and defoliation decreased nodule activity. Defoliated plants utilized nitrates in preference to dinitrogen fixation. Fertilizer N increased the concentration of nitrates in the plant, but the increase did not ameliorate the yield loss. Developing pods and seed are the predominate sink. The additional energy presumably required for dinitrogen fixation did not exacerbate the yield loss.     

Inhibition of low-density lipoprotein oxidation and up-regulation of low-density lipoprotein receptor in HepG2 cells by tropical plant extracts

Twelve edible plant extracts rich in polyphenols were screened for their potential to inhibit oxidation of low-density lipoprotein (LDL) in vitro and to modulate LDL receptor (LDLr) activity in cultured HepG2 cells. The antioxidant activity (inhibition of LDL oxidation) was determined by measuring the formation of conjugated dienes (lag time) and thiobarbituric acid reagent substances (TBARS). Betel leaf (94%), cashew shoot (63%), Japanese mint (52%), semambu leaf (50%), palm frond (41%), sweet potato shoot, chilli fruit, papaya shoot, roselle calyx, and maman showed significantly increased lag time (>55 min, P < 0.05) and inhibition of TBARS formation (P < 0.05) compared to control. LDLr was significantly up-regulated (P < 0.05) by Japanese mint (67%), semambu (51%), cashew (50%), and noni (49%). Except for noni and betel leaf, most plant extracts studied demonstrated a positive association between antioxidant activity and the ability to up-regulate LDL receptor. Findings suggest that reported protective actions of plant polyphenols on lipoprotein metabolism might be exerted at different biochemical mechanisms.