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Biochemical characterization of sap (latex) of a few Indian mango varieties.

Mango sap (latex) from four Indian varieties was studied for its composition. Sap was separated into non-aqueous and aqueous phases. Earlier, we reported that the non-aqueous phase contained mainly mono-terpenes having raw mango aroma (Phytochemistry 52 (1999) 891). In the present study biochemical composition of the aqueous phase was studied. Aqueous phase contained little amount of protein (2.0-3.5 mg/ml) but showed high polyphenol oxidase (147-214 U/mg protein) and peroxidase (401-561 U/mg protein) activities. It contained low amounts of polyphenols and protease activities. On native PAGE, all the major protein bands exhibited both polyphenol oxidase and peroxidase activities. Both polyphenol oxidase and peroxidase activities were found to be stable in the aqueous phase of sap at 4 degrees C. Sap contained large amount of non-dialyzable and non-starchy carbohydrate (260-343 mg/ml sap) which may be responsible for maintaining a considerable pressure of fluid in the ducts. Thus, the mango sap could be a valuable by-product in the mango industry as it contains some of the valuable enzymes and aroma components.

Carbohydrate Metabolism↗

Evolution of angiosperms via modulation of antagonisms.

The controversies concerning the evolutionary mechanisms of flowering plants continue unabated in spite of the current trends toward the analysis of macromolecular data and of the growing body of distributional micromolecular data. The usual narrative approach to this latter effort is here replaced by a novel technique, quantitative phytochemistry. An evolutionary outline emerges and reveals modulation of antagonisms as the fundamental mechanism of angiosperm evolutionary ecology. Origin or operation of many systems can be rationalized analogously. It is concluded that the impact of opposing features possesses universal relevance.

Biological Evolution↗

Purification and characterization of a protein phosphatase from winged bean.

A protein phosphatase (WbPP) has been purified from the soluble fraction of the winged bean (Psophocarpus tetragonolobus) shoot extract. The preparation is essentially homogenous as shown by the constant specific activity of the enzyme across the peak fractions, eluted from the thiophosphorylated histone-Sepharose affinity column, the last step of purification and by single protein bands on polyacrylamide gel electrophoresis (PAGE) in the presence as well as absence of denaturating agents. The monomeric nature of WbPP is revealed by an M(r) of 92,000 and 85,000, respectively, as estimated by SDS-PAGE and gel permeation chromatography under non-denaturating conditions. Autophosphorylated calmodulin-like domain protein kinase (P-WbCDPKI) [Saha, P., & Singh, M. (1995). Biochem. J., 305, 205] and phosphohistone H1 (P-hisH1), prepared by using the other homologous CDPK, i.e. WbCDPKII [Ganguly, S., & Singh, M. (1998). Phytochemistry, 48(1), 61], are good substrates of the purified enzyme, while P-hisH1 and phosphocasein prepared by using heterologous cAMP-dependent protein kinase, are respectively very poor and totally inactive as substrate. WbPP is adjudged to be a protein phosphoserine phosphatase since phosphoserine is the only phosphorylated amino acid residue detected in our earlier analysis of P-WbCDPKI and P-hisH1. The enzyme is strongly stimulated by a combination of Mg2+ and Ca2+, without being dependent on either of them and is also unaffected by calmodulin and fluphenazine. Orthovanadate strongly inhibits the enzyme while okadaic acid is a poor inhibitor.

Chromatography, Affinity↗

Chemotaxonomy and geographical distribution of tropane alkaloids.

This review illustrates the distribution of tropane alkaloids within the families Solanaceae, Erythroxylaceae, Proteaceae, Euphorbiaceae, Rhizophoraceae, Convolvulaceae and Cruciferae. Whereas tropane alkaloids are characteristic of the genera Datura, Brugmansia (tree datura) and Duboisia of the Solanaceae, the distribution is more widespread with novel tropane derivatives in families not traditionally associated with these bases. The chemical nature of more recently discovered water-soluble calystegines and the di- and trimeric forms from the Convolvulaceae (e.g. schizanthines from Schizanthus spp.), truxillines from Bolivian coca leaves and moonines of Erythroxylum moonii are highlighted. Where possible and appropriate, links between the phytochemistry and taxonomy are discussed.

Brassicaceae↗

10-Oxo-trans-8-decenoic acid (ODA): production, biological activities, and comparison with other hormone-like substances in Agaricus bisporus*

The few well-characterized fungal growth-regulating substances include 10-oxo-trans-8-decenoic acid (ODA) and hercynine. This report deals with production and tissue location of ODA. It also describes some biological activities of addition of ODA, hercynine, and cytokinins on growth and postharvest morphogenesis of Agaricus bisporus. Production of ODA in sporophore extracts was limited mainly by oxygen availability and the possible occurrence of a competitive metabolic pathway. Presumably synthesized within the stipe and skin tissues, ODA accumulated in the gills. Mycelium growth rate on a potato-based medium was significantly increased in the presence of ODA. Moreover, stipe lengthening was slightly stimulated by 10 or 100 µM ODA. Although these findings were similar to previous ones (Mau JL, Beelman RB, Ziegler GR. Phytochemistry 1992;31:4059-64), ODA appeared poorly active in our assays and mycelium growth on asparagine-glucose medium was strongly inhibited by 200 µM ODA. In contrast with cytokinins or hercynine, ODA did not speed up cap opening. Finally, tests carried out on animal cells suggested a dose-dependent cytotoxic effect of ODA.

Journal Article↗

The lectin from the mushroom Pleurotus ostreatus: a phosphatase-activating protein that is closely associated with an alpha-galactosidase activity. A part of this paper has been presented as a preliminary report at the 17th Interlec. Meeting 1997 in Würzburg, Germany.

The lectin from the mushroom Pleurotus ostreatus described earlier [F. Conrad, H. Rüdiger, The lectin from Pleurotus ostreatus: purification, characterization and interaction with a phosphatase, Phytochemistry 36 (1994) 277-283] was further characterized. Determination of the isoelectric point by capillary electrophoresis gave a value of 7.6. The dissociation constant of the lectin-alpha-lactose-1-phosphate complex determined by capillary electrophoresis is 3 mM. The activation of an endogenous phosphatase by the lectin as found earlier for the pseudosubstrate p-nitrophenylphosphate was confirmed also for naturally occurring substrates as ADP and ATP. We observed that at all purification steps the lectin is accompanied by an alpha-galactosidase activity. Both activities could neither be resolved by electrophoresis under non-denaturing conditions nor by affinity chromatography. However, carbohydrate binding by the lectin and carbohydrate processing by the enzyme are not due to the same site since: (i) the lectin accepts both alpha- and beta-glycosides whereas the enzyme activity is restricted to the alpha-anomer; (ii) the interaction with erythrocytes leads to a stable agglutinate, i.e. no 'clot-dissolving activity' [C.N. Hankins, J.I. Kindinger, L.M. Shannon, Legume alpha-galactosidases which have hemagglutinin properties, Plant Physiol. 65 (1980) 618-622] is observed; (iii) the alpha-galactosidase activity is inhibited by galactose but not by a beta-galactoside. Therefore, lectin and enzymatic activities are either properties of two tightly associated proteins, or of just one molecule. The kinetic parameters of the lectin-associated alpha-galactosidase activity for p-nitrophenyl-alpha-galactopyranoside are: K(M)=2.5 mM, k(cat)=66 s(-1), and K(I)=20mM for the inhibitor D-galactose.

Journal Article↗

Lantana camara L. (Verbenaceae).

Lantana camara L. is regarded both as a notorious weed and a popular ornamental garden plant and has found various uses in folk medicine in many parts of the world. Some taxa of the widely variable L. camara complex are toxic to small ruminants and this effect has been associated with the types and relative amounts of some triterpene ester metabolites. However, L. camara also produces a number of metabolites in good yields and some have been shown to possess useful biological activities. All these aspects are considered in this review to allow an evaluation of the potential for utilisation of the large biomass of Lantana available. The phytochemistry of other members of the Lantana genus is included.

Flavonoids↗

Medical ethnobotany of the Zapotecs of the Isthmus-Sierra (Oaxaca, Mexico): documentation and assessment of indigenous uses.

The Zapotec inhabitants of the Sierra de Oaxaca foothills (Mexico) live in an area of great botanic diversity. In daily subsistence and in response to illness, plants play a major role. An inventory of the Zapotec medicinal ethnobotany was carried out during 17 months of fieldwork. A total of 3611 individual responses concerning medicinal and non-medicinal uses for 445 different species of plants were documented. For the subsequent semi-quantitative analysis of data, the uses were grouped into ten categories and the responses for each species were summed up in each of these ten groups to yield rank-ordered lists. For the high rank-ordered and, hence, culturally important species, an assessment of the therapeutic potential was conducted using ethnobotanical, phytochemical and pharmacological data in the literature. Studies confirming the attributed properties or a scientific explanation of therapeutic use, as well as toxicological data, are still lacking for many of these species. The quantitative approach described will be the basis for future studies on the pharmacology and phytochemistry of Zapotec medicinal species. Finally, these data should also serve as a basis for biodiversity conservation and community development.

Ethnobotany↗

Oldenlandia affinis (R&S) DC. A plant containing uteroactive peptides used in African traditional medicine.

A review of the geographical distribution, clinical use, biological activity and phytochemistry of Oldenlandia affinis (R&S) DC. is presented. During an inventory of medicinal plants in northern Congo/Brazzaville and south-western Central African Republic in 1962, 196 different species were registered, one of which was O. affinis used for the facilitation of childbirth. A medical team working in Luluabourg (Kananga) in Congo during the troubled period in 1960, discovered also the traditional use of the same plant as an oxitocic agent during labour. The plant was collected and the uterotonic substances isolated. Cyclic peptides (called Kalata-peptides) were described, and the main peptide, B1, was subjected to pharmacological and chemical investigations. Later the three-dimensional structure of the peptide was determined. Similar cyclic peptides have been isolated also from other plants in the Rubiaceae family like Chassalia pasvifoloia and Psychotria longipes, and from Viola species: Viola tricolor L. and Viola arvensis Murray. Some of these peptides, included Kalata-peptide B1, have been shown to hold antimicrobial activity. They have recently been synthesized, and they may represent a starting point for the design of new peptide antibiotics.

Animals↗

Analysis and purification of phorbol esters using normal phase HPLC and photodiode-array detection.

For the first time a normal-phase HPLC method using photodiode-array detection is described for the analysis and purification of phorbol esters. The use of the method is demonstrated with examples of 10 different tigliane and daphnane esters (TPA, DOPP, DOPPA, Sap A, Sap B, Sap C, Sap D, Thy A, Ro and Rx). Both analytical and semipreparative techniques were developed. The method has been used in the final purification of DOPP and Rx from plant extracts. The method can be employed in the areas of phytochemistry, biochemistry and pharmacology/toxicology, where small samples of the toxic materials are required for research.

Chromatography, High Pressure Liquid↗

Anti-amnestic activity of E-p-methoxycinnamic acid from Scrophularia buergeriana.

We previously reported that phenylpropanoids isolated from the roots of Scrophularia buergeriana Miquel (Scrophulariaceae) protected cultured cortical neurons against glutamate-induced neurotoxicity [Kim and Kim, Phytochemistry, 54 (2000) 503-509; Kim et al., Br. J. Pharmacol. 135 (2002) 1281-1291]. In the present study, we examined the anti-amnestic activities of phenylpropanoids in mice with amnesia induced in vivo by scopolamine. Among the phenylpropanoids tested through passive avoidance tasks, buergeriside A1, buergeriside C1, E-p-methoxycinnamic acid (E-p-MCA) and E-isoferulic acid significantly improved the deficit of memory induced by scopolamine. This suggested that the alpha,beta-unsaturated carboxyl moiety and the para-methoxy group in phenylpropanoids (E-p-MCA) might be a crucial component in their cognition-enhancing activity. Indeed, E-p-MCA (0.01-2 mg/kg body weight, i.p.), given in pre- or post-treatment paradigms, significantly ameliorated scopolamine-induced amnesia as determined by passive avoidance tasks and prevented or aided in the recovery of memory to a level that was about 60% of control. In addition, E-p-MCA (0.1-1.0 mg/kg body weight, i.p.) significantly improved impairments of spatial learning and memory induced by scopolamine; the compound reduced deficits in both long- and short-term memories as measured by the Morris water maze test. We suggest, therefore, that E-p-MCA may ultimately hold significant therapeutic value in alleviating certain memory impairments observed in dementia.

Amnesia↗

Salivary thiol oxidase activity of Rhodnius prolixus.

Cysteine and other thiol compounds can accelerate the unloading of nitric oxide (NO) from salivary nitrosyl-nitrophorins of the blood sucking bug Rhodnius prolixus. The dependence of NO unloading on cysteine concentration is biphasic, showing a maximum between 0.5 and 1 mM cysteine. The proposed mechanism of action for the unloading is a series of reactions where cysteine (at low concentrations) reacts with the heme group of nitrophorins to form cystine and superoxide. The superoxide then reacts with NO to form peroxynitrite, which decays to a mixture of nitrite and nitrate anions. At high cysteine concentrations, cysteine is converted to cystine and H2O and thus no removal of NO from nitrophorins is observed. The thiol oxidase activity of Rhodnius nitrophorins is similar to that observed before in plant peroxidases [Pichorner et al., Phytochemistry 31, 3371 (1992)]. The possible physiological significance of this reaction to probing and feeding by R. prolixus is discussed.

Animals↗

Isolation and characterization of a novel lectin from Dioclea lehmanni (Fabaceae) seeds.

Dioclea lehmanni seeds are known to contain two lectins [G. Pérez, M. Hernández, E. Mora, Isolation and characterization of a lectin from the seeds of Dioclea lehmanni, Phytochemistry 29 (1990) 1745-1749]. Taking advantage of the strong bond shown by one of them (Dioclea lehmanni lectin I) to Sephacryl S-200, it has been purified and characterized as a Glc/Man lectin very similar to Concanavalin A. In order to compare the properties of the two lectins, we purified the second one (Dioclea lehmanni lectin II) to homogeneity by gel filtration and hydrophobic chromatography. Dioclea lehmanni lectin II is a dimeric protein (58.4 kDa) with identical subunits with M(r) = 29,000. Its molecular properties, carbohydrate specificity, human and animal erythroagglutination pattern, amino acid composition and N-terminal sequence (27 residues) were determined. These features clearly distinguish lectin II from lectin I; therefore a second novel lectin is present in the seeds of Dioclea lehmanni.

Amino Acid Sequence↗

Fast screening of low molecular weight compounds by thin-layer chromatography and "on-spot" MALDI-TOF mass spectrometry.

Fast screening of low-MW compounds is performed by thin-layer chromatography (TLC) followed by direct on-spot matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification with nearly "matrix-free" mass spectra using an UV-absorbing ionic liquid matrix. Owing to minimal background ions from the proton donor triethylamine/alpha-cyano-4-hydroxycinnamic acid ionic liquid matrix, three arborescidine alkaloids, the anesthesics levobupivacaine and mepivacaine, and the antibiotic tetracycline were readily characterized most frequently by the MS detection of their protonated molecules. The technique is fast and sensitive, requires little sample preparation and manipulation, and is therefore suitable for fast screening with TLC separation and MS identification of low-MW compounds, with potential applications in areas such as phytochemistry, synthetic chemistry, and product manufacturing quality monitoring.

Alkaloids↗

Characterization of isoflavones and their conjugates in female rat urine using LC/MS/MS.

Isoflavone phytoestrogens found in soybeans are the most widely studied phytochemicals in human diets and soy infant formulas. The health benefits of the isoflavones daidzein and genistein have been reported, and concerns about potential adverse effects have also been raised. However, the results of direct analysis of isoflavones and their metabolites in biological fluids after consumption of soy-containing diets are scarce. This study describes an LC/MS/MS method for the analysis of isoflavones and their metabolites in the urine of female rats fed diets made with soy protein isolate. Five isoflavones (daidzein, genistein, glycitein, dihydrodaidzein, and O-desmethylangolensin) were identified by comparison with authentic standards. Seventeen conjugates of isoflavones were characterized in the urine, the most unusual being genistein 5-glucuronide and four glucuronide conjugates of reductive metabolites of daidzein. The application of LC/MS/MS to analyze isoflavone metabolites is simple and sensitive, and appears to be an excellent method for determining the bioavailability and metabolism of food phytochemistry.

Animals↗

Isolation and synthesis of allelochemicals from gramineae: benzoxazinones and related compounds.

Compounds with a (2H)-1,4-benzoxazin-3(4H)-one skeleton have attracted the attention of phytochemistry researchers since 2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one (DIBOA) and 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (DIMBOA) were isolated from plants belonging to the Poaceae family. These compounds exhibit interesting biological properties, such as phytotoxic, antimicrobial, antifeedant, antifungal, and insecticidal properties. These chemicals, in addition to a wide variety of related compounds involved in their metabolism, detoxification mechanisms, and degradation on crop soils and other systems, have high interest and in some cases potential agronomic utility. This paper presents a complete review of the methods employed for their synthetic obtention in addition to some of the authors' own contributions to their chemistry. The degradation and phytotoxicity experiments carried out in ongoing research into the potential agronomic utility of these compounds required large amounts of them, which were obtained from natural sources. This paper presents a modified methodology to access DIMBOA from Zea mays cv. Apache and to obtain 2-O-beta-D-glucopyranosyl-2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one (DIBOA-Glc) and DIBOA from Secale cereale L. New synthetic methodologies were employed for the obtention of the lactams 2-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one and 2-hydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one and the malonamic acids N-(2-hydroxyphenyl)malonamic acid and N-(2-hydroxy-7-methoxyphenyl)malonamic acid. The aminophenoxazines 2-amino-7-methoxyphenoxazin-3-one and 2-acetamido-7-methoxyphenoxazin-3-one have been synthesized in the authors' laboratory by novel procedures. All of the methodologies employed allowed the desired compounds to be obtained in high yield and in an easy-to-scale manner.

Benzoxazines↗

Two new norditerpenoid alkaloids from Delphinium elatum var. "black night".

Two new norditerpenoid alkaloids, blacknine [1] and blacknidine [2], have been isolated from the whole plants of Delphinium elatum var. "black night" along with six known alkaloids. The structures of the new alkaloids 1 and 2 were derived from their spectroscopic data (1H, 13C, DEPT, COSY, HETCOR, and selective INEPT nmr experiments). The phytochemistry of this plant variety has not been examined previously.

Alkaloids↗

High performance liquid chromatography-mass spectrometry for metabonomics: potential biomarkers for acute deterioration of liver function in chronic hepatitis B.

Metabonomics methods have been successfully applied to the drug discovery, toxicology, phytochemistry, and clinical fields. Here, we report a self-developed metabonomics platform which is based on high performance liquid chromatography-mass spectrometry (HPLC-MS) technique and applied to the investigation of acute deterioration of liver function in chronic hepatitis B to find the potential biomarkers. Sera from 50 healthy persons and 37 patients with acute deterioration of liver function in chronic hepatitis B were analyzed by HPLC-MS after removal of proteins. After de-noise, peak detection and peak alignment, the data of metabolites were fed to partial least squares discriminant analysis (PLS-DA) to find the potential biomarkers. According to the corresponding tandem mass results, several potential biomarkers were identified: Lysophosphatidyl Choline (LPC) C18:0, LPC C16:0, LPC C18:1, LPC C18:2, and glycochenodeoxycholic acid (GCDCA) (or its isomer glycodeoxycholic acid (GDCA)). On the basis of the relevant literature and pathway databases, the biological significance of the present study is discussed.

Adult↗