Procyanidin C1 from Viola odorata L. inhibits Na+,K+-ATPase.

Members of the Viola genus play important roles in traditional Asian herbal medicine. This study investigates the ability of Viola odorata L. extracts to inhibit Na+,K+-ATPase, an essential animal enzyme responsible for membrane potential maintenance. The root extract of V. odorata strongly inhibited Na+,K+-ATPase, while leaf and seeds extracts were basically inactive. A UHPLC-QTOF-MS/MS metabolomic approach was used to identify the chemical principle of the root extract's activity, resulting in the detection of 35,292 features. Candidate active compounds were selected by correlating feature area with inhibitory activity in 14 isolated fractions. This yielded a set of 15 candidate compounds, of which 14 were preliminarily identified as procyanidins. Commercially available procyanidins (B1, B2, B3 and C1) were therefore purchased and their ability to inhibit Na+,K+-ATPase was investigated. Dimeric procyanidins B1, B2 and B3 were found to be inactive, but the trimeric procyanidin C1 strongly inhibited Na+,K+-ATPase with an IC50 of 4.5 µM. This newly discovered inhibitor was docked into crystal structures mimicking the Na3E1∼P·ADP and K2E2·Pi states to identify potential interaction sites within Na+,K+-ATPase. Possible binding mechanisms and the principle responsible for the observed root extract activity are discussed.

The Phenolics and Antioxidant Properties of Black and Purple versus White Eggplant Cultivars.

The total phenolic content, anthocyanins, phenolic acids, antioxidant capacity and α-amylase inhibitory activity of black (Aydin Siyahi), purple (Kadife Kemer) and white (Trabzon Kadife) eggplants grown in Turkey were subjected to a comparative investigation. The black cultivar exhibited the highest total phenolic (17,193 and 6552 mg gallic acid equivalent/kg fw), flavonoid (3019 and 1160 quercetin equivalent/kg fw) and anthocyanin (1686 and 6167 g delphinidin-3-O-glucoside equivalent/kg fw) contents in crude extracts of the peel and pulp. The majority of the caffeic acid was identified in the ester (2830 mg/kg fw) and ester-bound (2594 mg/kg fw) forms in the peel of 'Kadife Kemer' and in the glycoside form (611.9 mg/kg fw) in 'Aydin Siyahi', as well as in the pulp of these two eggplants. 'Kadife Kemer' (purple eggplant) contained the majority of the chlorogenic acid in free form (27.55 mg/kg fw), compared to 'Aydin Siyahi' in the ester (7.82 mg/kg fw), glycoside (294.1 mg/kg dw) and ester-bound (2.41 mg/kg fw) forms. The eggplant cultivars (peel and pulp, mg/kg fw) exhibited a relatively high delphinidin-3-O-rutinoside concentration in the peel of 'Aydin Siyahi' (avg. 1162), followed by 'Kadife Kemer' (avg. 336.6), and 'Trabzon Kadife' (avg. 215.1). The crude phenolic extracts of the eggplants exhibited the highest antioxidant capacity values (peel and pulp, µmoL Trolox equivalent/kg fw) of 2,2-diphenyl-1-picrylhydrazyl (DPPH, 8156 and 2335) and oxygen radical absorbance capacity (ORAC, 37,887 and 17,648). The overall results indicate that black and purple eggplants are the cultivars with greater potential benefits in terms of their phenolics and antioxidant values than the white eggplant.

Antibacterial, Mutagenic Properties and Chemical Characterisation of Sugar Bush (Protea caffra Meisn.): A South African Native Shrub Species.

Protea caffra is used as a diarrhoeal remedy in South African herbal medicine, however, its pharmacological properties remain largely unknown. In the present study, extracts from different Protea caffra organs were screened against drug-sensitive and -resistant diarrhoeagenic pathogens using the microdilution assay (minimum inhibitory concentration, MIC). Twig extracts (70% methanol, MeOH) of the plant were purified and the resultant fractions screened for antibacterial properties (MIC). The chemical profiles of the fractions were determined by Gas Chromatography-Mass Spectrometry (GC-MS), while ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to quantify the phenolic acids in the plant. The mutagenic properties of bioactive extracts were assessed using the Ames test. The extracts demonstrated weak-moderate antibacterial properties (MIC: 0.3-0.6 mg/mL). A cold ethyl acetate fraction of MeOH twig extract exhibited significant antibacterial properties (MIC = 0.078 mg/mL) against Enterococcus faecalis. The presence of antibacterial compounds (1-adamantane carboxylic acid, heptacosanol, levoglucosan, nonadecanol) in the plant was putatively confirmed based on GC-MS analysis. Furthermore, UHPLC-MS/MS analysis revealed varying concentrations of phenolic acids (0.08-374.55 µg/g DW). Based on the Ames test, the extracts were non-mutagenic thereby suggesting their safety. To a certain degree, the current study supports the traditional use of Protea caffra to manage diarrhoea among local communities in South Africa.

Phytochemical Profiles and Antioxidant Activity of Grasses Used in South African Traditional Medicine.

Grasses are a valuable group of monocotyledonous plants, used as nourishing foods and as remedies against diseases for both humans and livestock. Phytochemical profiles of 13 medicinal grasses were quantified, using spectrophotometric methods and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS), while the antioxidant activity was done using DPPH and ferric-reducing-power assays. The phytochemical analysis included the total soluble phenolic content, flavonoids, proanthocyanidins, iridoids and phenolic acids. Among the 13 grasses, the root methanolic extracts of Cymbopogon spp., Cymbopogon nardus and Cenchrus ciliaris contained the highest concentrations of total soluble phenolics (27-31 mg GAE/g DW) and flavonoids (4-13 mg CE/g DW). Condensed tannins and total iridoid content were highest (2.3 mg CCE/g DW and 3.2 mg HE/g DW, respectively) in Cymbopogon nardus. The most common phenolic compounds in the grass species included ρ-coumaric, ferulic, salicylic and vanillic acids. In the DPPH radical scavenging assay, the EC50 values ranged from 0.02 to 0.11 mg/mL for the different grasses. The best EC50 activity (lowest) was exhibited by Cymbopogon nardus roots (0.02 mg/mL) and inflorescences (0.04 mg/mL), Cymbopogon spp. roots (0.04 mg/mL) and Vetiveria zizanioides leaves (0.06 mg/mL). The highest ferric-reducing power was detected in the whole plant extract of Cynodon dactylon (0.085 ± 0.45; r2 = 0.898). The observed antioxidant activity in the various parts of the grasses may be due to their rich pool of phytochemicals. Thus, some of these grasses provide a source of natural antioxidants and phytochemicals that can be explored for their therapeutic purposes.

Potential of Smoke-Water and One of Its Active Compounds (karrikinolide, KAR1) on the Phytochemical and Antioxidant Activity of Eucomis autumnalis.

Eucomis autumnalis (Mill.) Chitt. subspecies autumnalis is a popular African plant that is susceptible to population decline because the bulbs are widely utilized for diverse medicinal purposes. As a result, approaches to ensure the sustainability of the plants are essential. In the current study, the influence of smoke-water (SW) and karrikinolide (KAR1 isolated from SW extract) on the phytochemicals and antioxidant activity of in vitro and greenhouse-acclimatized Eucomis autumnalis subspecies autumnalis were evaluated. Leaf explants were cultured on Murashige and Skoog (MS) media supplemented with SW (1:500, 1:1000 and 1:1500 v/v dilutions) or KAR1 (10-7, 10-8 and 10-9 M) and grown for ten weeks. In vitro regenerants were subsequently acclimatized in the greenhouse for four months. Bioactive phytochemicals in different treatments were analyzed using ultra-high performance liquid chromatography (UHPLC-MS/MS), while antioxidant potential was evaluated using two chemical tests namely: DPPH and the ß-carotene model. Smoke-water and KAR1 generally influenced the quantity and types of phytochemicals in in vitro regenerants and acclimatized plants. In addition to eucomic acid, 15 phenolic acids and flavonoids were quantified; however, some were specific to either the in vitro regenerants or greenhouse-acclimatized plants. The majority of the phenolic acids and flavonoids were generally higher in in vitro regenerants than in acclimatized plants. Evidence from the chemical tests indicated an increase in antioxidant activity of SW and KAR1-treated regenerants and acclimatized plants. Overall, these findings unravel the value of SW and KAR1 as potential elicitors for bioactive phytochemicals with therapeutic activity in plants facilitated via in vitro culture systems. In addition, it affords an efficient means to ensure the sustainability of the investigated plant. Nevertheless, further studies focusing on the use of other types of antioxidant test systems (including in vivo model) and the carry-over effect of the application of SW and KAR1 for a longer duration will be pertinent. In addition, the safety of the resultant plant extracts and their pharmacological efficacy in clinical relevance systems is required.

Identification of Narciclasine as an in Vitro Anti-Inflammatory Component of Cyrtanthus contractus by Correlation-Based Metabolomics.

In this study, an extract from the bulbs of Cyrtanthus contractus showed strong anti-inflammatory activity in vitro. The extract was partially separated into 14 fractions and analyzed by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry metabolomics, and the correlation coefficients were calculated between biological activities and metabolite levels. As a result, the top-scoring metabolite narciclasine (1) is proposed as the active principle of C. contractus. This was confirmed by comparing the biological effect of crude extract with that of an authentic standard.

Identification and characterization of potential bioactive compounds from the leaves of Leucosidea sericea.

ETHNOPHARMACOLOGICAL RELEVANCE: Leucosidea sericea is a southern African tree used for treating different diseases including microbial infections and inflammatory-related conditions. AIM OF THE STUDY: To isolate and identify the chemicals in Leucosidea sericea which possibly account for the diverse therapeutic effects of the species. MATERIALS AND METHODS: Leaf material was extracted using 20% methanol and subsequently partitioned with different solvents (hexane, dichloromethane, ethyl acetate and butanol). Resultant fractions were subjected to bioactive (antimicrobial)-guided isolation and the structural elucidation was conducted using NMR (1D and 2D) spectroscopic methods. Given the broad uses of Leucosidea sericea in traditional medicine, the extract, fractions and isolated compounds were evaluated in five (5) biological assays in vitro (antimicrobial, antioxidant, acetylcholinesterase (AChE) and anti-inflammatory inhibition as well as cytotoxicity effect). RESULTS: As the most active fractions, from ethyl acetate yielded 5,7-dihydroxychromone (1); 1-hydroxy-2-oxopomolic acid (2); 3,5,7,3´,4´-pentahydroxyflavone (3) and Tiliroside (4). For the first time, these four (4) compounds were isolated from leaves of Leucosidea sericea. These aforementioned compounds demonstrated broad-spectrum antibacterial activity (1.95-125 µg/mL) and noteworthy antifungal (3.9-250 µg/mL) potential. In addition to its noteworthy antimicrobial activity, compound 3 also demonstrated significant antioxidant (EC50 = 14 µg/mL in DPPH assay) and anti-inflammatory (inhibited the level of ELAM by approximately 36% and decreased also the viability of endothelial cells) activities in vitro. Overall, AChE inhibition activity and cytotoxic response was generally weak for the extracts, fractions and isolated compounds. CONCLUSION: The pool of chemicals in Leucosidea sericea were enriched with the isolation and identification of four (4) compounds obtained from the leaf extract. Among these compounds, the significant antimicrobial activity of compound 3 provides strong evidence that support the use of Leucosidea sericea for microbial-related infections in folk medicine.

Metabolite profiling and isolation of biologically active compounds from Scadoxus puniceus, a highly traded South African medicinal plant.

Scadoxus puniceus (Amaryllidaceae), a medicinal plant of high value in South Africa, is used as a component of a traditional herbal tonic prescribed to treat several ailments. Ultra-high performance liquid chromatography-tandem mass spectrometry quantified the phenolic compounds in different organs of S. puniceus. Gravity column chromatography was used to separate fractions and active compounds. The structure of these compounds was determined using 1D and 2D nuclear magnetic resonance and mass spectroscopic techniques. A microplate technique was used to determine the acetylcholinesterase inhibitory activity of the pure compounds. Metabolite profiling revealed a greater profusion of hydroxycinnamic acids (69.5%), as opposed to hydroxybenzoic acids (30.5%). Chlorogenic acid was the most abundant (49.6% of hydroxycinnamic acids) compound. In addition to chlorogenic acid, the study is the first to report the presence of sinapic, gallic, and m-hydroxybenzoic acids in the Amaryllidaceae. Chromatographic separation of S. puniceus led to the isolation of haemanthamine (1), haemanthidine (2), and a rare chlorinated amide, metolachlor (3), the natural occurrence of which is described for the first time. Haemanthamine, haemanthidine, and metolachlor displayed strong acetylcholinesterase inhibitory activity (IC50 ; 23.1, 23.7, and 11.5 µM, respectively). These results substantiate the frequent use of S. puniceus as a medicinal plant and hold much promise for further pharmaceutical development.

Antifungal and antioxidant activities of Coleonema album and C. pulchellum against skin diseases.

CONTEXT: Coleonema album (Thunb) Bart. & H. L. Wendl (Rutaceae) has been used in the formulation of skincare products, and the Khoisan people rub it on their skin to add luster. Coleonema pulchellum I. Williams has received less attention in the South African traditional medicine. OBJECTIVE: This study investigates the antifungal and antioxidant activities of C. album and C. pulchellum essential oil (EO) and leaf extracts; and analyzes the chemical components of their EOs. MATERIALS AND METHODS: Antifungal activity of leaf extracts was determined using the microdilution method with griseofulvin and ketoconazole as controls. Antifungal capacity of EO was investigated using the 'Volatile release plate method'. Trichophyton rubrum (ATCC 28188) and T. mentagrophytes (ATCC 9533) mycelia (0.3 cm diameter) were placed on fresh yeast malt agar in Petri dishes with filter paper (impregnated with 20 µL of EO) on the lid for direct exposure to EO volatiles while plates without EO were used as controls. The incubation time was seven days. Antioxidant activities of the leaf extracts were determined. RESULTS: Methanol leaf extract of C. pulchellum inhibited the growth of three fungi tested with MIC values of 195, 391 and 49 µg/mL for Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum gypseum, respectively. Terpenes formed the major components of the EO. The EO from both plants inhibited the growth of T. rubrum in vitro. DISCUSSION AND CONCLUSION: This study revealed the therapeutic value of C. pulchellum. Coleonema album and C. pulchellum should be considered as potential plants for skin ointment from natural origin.

Phytochemical Characterization, Antibacterial, Acetylcholinesterase Inhibitory and Cytotoxic Properties of Cryptostephanus vansonii, an Endemic Amaryllid.

Cryptostephanus vansonii I. Verd., an endemic Amaryllidaceae species from Zimbabwe, was evaluated for its acetylcholinesterase (AChE) inhibitory and cytotoxicity properties using Ellman's colorimetric method and the tetrazolium-based colorimetric assay against Vero monkey kidney cells, respectively. The plant extracts were also evaluated for their antibacterial activity against five bacteria. Furthermore, phytochemical profiles of the extracts were determined using ultra-high performance liquid chromatography coupled with tandem mass spectrometry analysis. A plant part-dependent AChE inhibitory activity was observed, in the order, root > rhizome > basal leaf > leaf. Overall, C. vansonii extracts exhibited better antibacterial activity against Gram-negative compared with Gram-positive bacteria. Cytotoxic effects were not detected in Vero monkey kidney cell lines suggesting the possible absence of toxophores in C. vansonii extracts. Similar to the trend in biological activity, a distinct plant part-dependent variation in hydroxybenzoates, hydroxycinnamates and flavonoids was observed in the plant extracts. In addition, 5-hydroxymetylfurfural and eucomic acid were detected in the different plant parts of C. vansonii. The results of the present study provide valuable AChE inhibition activity, toxicological and phytochemical profiles of C. vansonii. Further studies on isolation of bioactive compounds and their subsequent evaluation in other pharmacological and toxicological model systems are required. Copyright © 2017 John Wiley & Sons, Ltd.

Bog bilberry phenolics, antioxidant capacity and nutrient profile.

Phenolics and nutrient profiles of bog bilberry (Vaccinium uliginosum L.) collected from high mountain pastures in northeast Anatolia (Turkey) were examined for the first time in this study. The major soluble sugar identified in the berry was fructose, following by glucose, and the main organic acid identified was citric acid, followed by malic acid. Eleven phenolic acids and 17 anthocyanin 3-glycosides were identified and quantified. Caffeic acid in the free and glycoside forms and syringic acid in the ester form were the major phenolic acids, and the major individual anthocyanin present in the berry was malvidin 3-glucoside (24%). The highest total phenolics and anthocyanin contents were obtained from the anthocyanin fraction in conjunction with the highest antioxidant capacity, followed by the polyphenolic and aqueous fractions, FRAP, ORAC and DPPH, in that order. Our findings can be used to compare bog bilberry with other Vaccinium berries and to help clarify the relative potential health benefits of different berries.

Content of metals and metabolites in honey originated from the vicinity of industrial town Kosice (eastern Slovakia).

Composition of three types of honey (mixed forest honey and monofloral-black locust and rapeseed honeys) originated from the vicinity of an industrial town (Kosice, Slovak Republic) was compared. Higher content of minerals including toxic metals in forest honey (1358.6 ng Ni/g, 85.6 ng Pb/g, and 52.4 ng Cd/g) than in rapeseed and black locust honeys confirmed that botanical origin rather than the distance for eventual source of pollution (steel factory) affects metal deposition. Benzoic acid derivatives were typically more accumulated in forest but cinnamic acid derivatives and some flavonoids in rapeseed honey (in free and/or glycoside-bound fraction). In terms of quantity, p-hydroxybenzoic and p-coumaric acids were mainly abundant. Total phenols, thiols, and proteins were abundant in forest honey. Some metals and phenols contributed to separation of honeys based on principal component analysis (PCA). Native amount of 5-(hydroxymethyl)furfural was not related to honey type (~11 µg/g) and was elevated after strong acid hydrolysis (200-350 µg/g) but it did not interfere with the assay of phenols by Folin-Ciocalteu reagent. This is the first report of metals and metabolites in the same study, and data are discussed with available literature. We conclude that black locust (acacia) honey is the most suitable for daily use and that central European monofloral honeys contain lower amounts of toxic metals in comparison with other geographical regions.

Antimicrobial, Anthelmintic Activities and Characterisation of Functional Phenolic Acids of Achyranthes aspera Linn.: A Medicinal Plant Used for the Treatment of Wounds and Ringworm in East Africa.

Achyranthes aspera Linn. (Amaranthaceae) commonly known as Prickly Chaff flower (English) is traditionally used for treating a number of ailments. Different parts of the plant are used in treating wounds and ringworm in East Africa and elsewhere for a number of ailments. In this study, leaf extracts of A. aspera collected from two different geographical locations (Ciaat, Eritrea and Ukulinga, South Africa) were evaluated for antibacterial, antifungal, anthelmintic activities and the plant characterized for functional phenolic acids as well as protein binding capacity. The pathogens used in the tests were, two Gram-negative (Escherichia coli and Klebsiella pneumoniae), two Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), a filamentus yeast-like fungus (Candida albicans) and a free-living nematode (Caenorhabditis elegans). The water and acetone extracts of the samples collected from Ciaat exhibited good antibacterial, antifungal and anthelmintic activity (MIC < 1 mg/ml) except the water extract against E. coli which showed moderate activity. In contrast, the extracts collected from Ukulinga exhibited moderate to weak activities except for the acetone (aq.) extracts which had good activity against some of the tested organisms. UHPLC-MS/MS revealed variation in the levels of some functional phenolic compounds, with rutin, chlorogenic acid and genistein not being detected in the extracts from Ukulinga. The variation was also observed in the protein binding capacity, which could offer a predictive wound healing model. All extracts from plant samples collected at Ciaat expressed significant dominant potency compared to similar extracts from Ukulinga.

Genetic and phytochemical variability of six Teucrium arduini L. populations and their antioxidant/prooxidant behaviour examined by biochemical, macromolecule- and cell-based approaches.

Teucrium arduini L., an Ilyric-Balcanic endemic species, has been reported for decades as a valuable plant used in traditional medicine for treating digestive disorders. The present study evaluated genetic and phytochemical variability of six T. arduini populations in order to determine factors that influence an accumulation of polyphenolic compounds. Results strongly suggest that a phytochemical variation was caused by environmental rather than genetic factors. T. arduini leaf extract from the locality Ucka, which accumulated significantly more polyphenolic phytochemicals in comparison to others, showed antioxidant activity in DNA and lipid bioassays. Furthermore, the same extract exhibited prooxidant behaviour at protein level and induce formation of reactive oxygen species in human laryngeal carcinoma cells causing cytotoxic activity, in a dose dependent manner. All the results of the present study suggested that T. arduini extract could be responsible for antioxidative/prooxidative mechanisms and would help in determination of optimal conditions for their ethnopharmacological use.

Molecular and cellular approach in the study of antioxidant/pro-oxidant properties of Micromeria croatica (Pers.) Schott.

Phytochemical composition and antioxidant activity of three wild populations of endemic Illyric-Balcanic species Micromeria croatica (Pers.) Schott have been evaluated with respect to plant organ and growing location. Multivariate analysis (principal component analysis) was performed to visualise (dis-)similarity among samples and identify the correlations between phytochemical variables that explain the most variability. The tested leaf extract from BaCic kuk locality exhibited protective effects against reactive oxygen species-induced damage of DNA and inhibition of lipid peroxidation, while it caused oxidative degradation of protein in the bovine serum albumin assay at higher concentrations. This extract also exhibited cytotoxic activity and facilitated the formation of reactive oxygen species in the HEp2 cell line, in a dose-dependent manner.

Conservation strategy for Pelargonium sidoides DC: phenolic profile and pharmacological activity of acclimatized plants derived from tissue culture.

ETHNOPHARMACOLOGICAL RELEVANCE: Pelargonium sidoides DC (Geraniaceae), a popular medicinal plant used in folk medicine in the treatment of respiratory-related infections has gained international prominence due to its usage in several herbal formulations. This has led to high demand and the subsequent decimation of wild populations. AIM OF THE STUDY: Using plant tissue culture techniques, Pelargonium sidoides plants were cloned in vitro, acclimatized under greenhouse conditions and evaluated for their phytochemical content and pharmacological activity. METHODS: Phenolic content in extracts of in vitro-derived, greenhouse-acclimatized and wild Pelargonium sidoides plants were analyzed using UPLC-MS/MS. The oxygen radical absorbance capacity (ORAC), 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging activity and minimum inhibitory concentration (MIC) of the extracts against bacterial and fungal strains were evaluated. RESULTS: Similarities in phenolic profiles were identified confirming the chemical signatures that characterize Pelargonium sidoides plants. Extracts of greenhouse-acclimatized and wild plants exhibited comparable antimicrobial and antioxidant properties. CONCLUSIONS: Overall, the study highlights the potential of integrating plant tissue culture technologies in conservation strategies of medicinal plants. In particular, the results strongly suggest the feasibility of both large-scale cultivation and plant part substitution as alternative solutions to the current destructive overharvesting practices of wild Pelargonium sidoides populations.

Antioxidant and phenolic acid profiles of tissue cultured and acclimatized Merwilla plumbea plantlets in relation to the applied cytokinins.

Merwilla plumbea (Lindl.) Speta is an important medicinal plant widely used in traditional medicine. We evaluated the effect of five cytokinins [benzyladenine (BA), 2-isopentenyladenine (2iP), meta-topolin (mT), meta-topolin riboside (mTR), and meta-methoxy-9-tetrahydropyran-2-yl-topolin (MemTTHP)] on the level of phenolic acids and antioxidant activity of M. plumbea during the tissue culture and acclimatization stages. Two cytokinins (mT and mTR) significantly improved the antioxidant activity of tissue culture plantlets while the control plantlets were better after acclimatization. Using UPLC-MS/MS, the levels of hydroxybenzoic and hydroxycinnamic acid derivatives (phenolic acids) varied significantly during tissue culture and acclimatization, depending on the cytokinin and plant part analyzed. Vanillic acid (24.9 µg g⁻¹) detected in underground parts of tissue culture plants supplemented with BA was the most abundant phenolic acid detected. The current findings indicate that the phytochemicals together with the bioactivity during in vitro propagation of M. plumbea is influenced by the cytokinin type used and the stage of plant material collection.

Antiproliferative and antiangiogenic effects of flavone eupatorin, an active constituent of chloroform extract of Orthosiphon stamineus leaves.

Flavone eupatorin is one of the constituents of Orthosiphon stamineus, a medicinal herb used in folk medicine in South East Asia for treatment of various disorders. In our study, we investigated the antiproliferative properties of a chloroform extract of the leaves of O. stamineus and of pure eupatorin. The compound was able to reduce the number of viable cancer cells to the same extent as the extract, with IC(50) values in micromolar range. Moreover, both the eupatorin standard and the extract caused cells to arrest in the G2/M phase of the cell cycle. This clearly demonstrates that eupatorin contributes significantly to the overall extract activity. Induction of mitotic catastrophe, accompanied by key molecular events defining apoptosis, is the mechanism of eupatorin-induced cell death. Importantly, eupatorin (at the doses cytotoxic to cancer cells) did not kill normal cells; it only limited migration of HUVEC endothelial cells and their ability to create tubes. The ability of eupatorin to nonspecifically inhibit many protein kinases was proven and is the probable cause of its cellular effects. In summary, eupatorin emerges as a promising agent in anticancer research.

Phytochemical and antimicrobial characterization of Macleaya cordata herb.

Macleaya cordata (plume poppy) is a source of bioactive compounds, mainly isoquinoline alkaloids which are used in phytopreparations with anti-inflammatory and antimicrobial activities. In this study, the alkaloids sanguinarine, chelerythrine, their dihydro derivatives, protopine and allocryptopine and phenolics, gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, p-coumaric, caffeic, ferulic and sinapic acids were determined in extracts prepared from M. cordata aerial part, seeds, and seed capsules using HPLC with UV detection and/or LC/MS with electrospray ionization. The highest content of sanguinarine and chelerythrine was found in capsules. Protopine and allocryptopine were major alkaloids in leaves including footstalks. The seed oil contained dihydrosanguinarine, dihydrochelerythrine and twelve fatty acids of which linoleic, oleic, palmitic and stearic acids predominated. In addition, sanguinarine reductase, a key enzyme in sanguinarine/dihydrosanguinarine equilibrium in plants, was found for the first time, in the soluble proteins of leaves. Finally, extracts were tested for antimicrobial activity using the microdilution method on standard reference bacterial strains.

Cadmium and nickel uptake are differentially modulated by salicylic acid in Matricaria chamomilla plants.

Chamomile (Matricaria chamomilla) is a widely used medicinal plant which also accumulates heavy metals in its above-ground organs. We investigated the effect of the important plant signaling molecule, salicylic acid (SA), on the accumulation of Ni or Cd, by exposing plants over 7 days to 60 microM solutions of individual heavy metals with or without 50 microM SA. Special emphasis was focused on phenolic metabolism-related parameters, not only because of their importance for growth and stress tolerance but also because phenolics are potent antioxidants in human diet. In combined treatments, SA stimulated an increase in soluble proteins of roots and reduced their water content. SA reduced total Cd in the shoot and increased Ni. Total and "intraroot" Ni decreased in Ni + SA treatment, while in the case of Cd, only "intraroot" content decreased in Cd + SA treatment, being correlated with cell wall-bound phenolic acids and lignin. SA was strongly accumulated in roots from the Ni + SA treatment, being correlated with an increase in hydrogen peroxide. In both Cd + SA and Ni + SA treatments, SA enhanced phenylalanine ammonia-lyase activity and accumulation of total soluble phenols, particularly in the roots. Here, we report for the first time that soluble phenols may be involved in Cd shoot-to-root translocation. In the case of Ni, it seems that phenols serve as a root barrier in order to prevent Ni from reaching the above-ground organs. The effects of SA on phenolic metabolism, and the signaling role of ROS in the accumulation of phenols, are discussed.