Lythrum salicaria Ellagitannins Stimulate IPEC-J2 Cells Monolayer Formation and Inhibit Enteropathogenic Escherichia coli Growth and Adhesion.

Lythrum salicaria herb (LSH) was applied in diarrhea therapy since ancient times. Despite empirically referenced therapeutic effects, the bioactivity mechanisms and chemical constituents responsible for pharmacological activity remain not fully resolved. Taking into consideration the historical use of LSH in treatment of diarrhea in humans and farm animals, the aim of the study was to examine in vitro the influence of LSH and its C-glycosylic ellagitannins on processes associated with maintaining intestinal epithelium integrity and enteropathogenic Escherichia coli (EPEC) growth and adhesion. LSH was not only inhibiting EPEC growth in a concentration dependent manner but also its adhesion to IPEC-J2 intestinal epithelial cell monolayers. Inhibitory activity toward EPEC growth was additionally confirmed ex vivo in distal colon samples of postweaning piglets. LSH and its dominating C-glycosylic ellagitannins, castalagin (1), vescalagin (2), and salicarinins A (3) and B (4) were stimulating IPEC-J2 monolayer formation by enhancing claudin 4 production. Parallelly tested gut microbiota metabolites of LSH ellagitannins, urolithin C (5), urolithin A (6), and its glucuronides (7) were inactive. The activities of LSH and the isolated ellagitannins support its purported antidiarrheal properties and indicate potential mechanisms responsible for its beneficial influence on the intestinal epithelium.

Endotoxin Contamination and Reaction Interfering Substances in the Plant Extract Library.

Endotoxin is an unintentional contaminant that has numerous activities and can affect various biological experiments using cells. In this study, we measured the endotoxin activity of samples from a plant extract library (PEL) and determined their degrees of contamination. Endotoxin was detected in approx. 48% (n = 139) and approx. 4% (n = 5) of field-collected and crude drug samples, respectively, and in concentrations >5.0 EU/mL in some samples. The concentrations of endotoxin that affect cells in vitro vary depending on the target cell type. Although the degree of contamination varied in the present study, it was considered to have little effect on the cell experiments. More than 150 PEL samples had problems with reaction courses or recovery rates of Limulus amoebocyte lysate (LAL) tests. In the LAL tests, using three plant extracts [Sanguisorba officinalis L. (Rosaceae), Oenothera biennis L. (Onagraceae), and Lythrum salicaria L. (Lythraceae)], the polyphenolic compounds in the plant extracts affected LAL test and their effects differed depending on the plant species. When the 16 single polyphenol compounds were added to the LAL tests, the compounds with caffeoyl and pyrogallol moieties were found to affect the LAL reaction course and recovery rate. Furthermore, none of the compounds had any effects at concentrations of 1 µM. Because the plant extracts contained analogs of various polyphenolic compounds, they were presumed to actually act synergistically. Our findings demonstrated that attention must be paid to the recovery rate and reaction process of LAL tests with samples containing polyphenolic compounds.

Lythrum salicaria L. herb and gut microbiota of healthy post-weaning piglets. Focus on prebiotic properties and formation of postbiotic metabolites in ex vivo cultures.

ETHNOPHARMACOLOGICAL RELEVANCE: Herb of purple loosestrife (Lythrum salicaria L. from Lythraceae family) (LSH) was used in Europe since ancient times till early-20th century in the therapy of diarrhea and dysentery in human and veterinary medicine. Post-weaning diarrhea is a main problem affecting global piglet production, which leads to significant economic losses because of increased morbidity and mortality, reduced average daily gain, and high antibiotic consumption. Post-weaning diarrhea has various causes, all of which have been linked to imbalances of intestinal microbiota. The aim of the present study was to determine the interaction of LSH with the gut microbiota of healthy post-weaning piglets in order to evaluate its influence on microbiota composition and metabolism as well as production of potentially bioactive postbiotic metabolites from the extract constituents. MATERIALS AND METHODS: Ex vivo anaerobic cultures of piglets intestinal microbiota obtained from jejunum, ileum, caecum and distal colon were conducted in various culture media supplemented with LSH. The production of postbiotic metabolites was determined using UPLC-DAD-MSn method. Bacterial genomic DNA was extracted and examined by sequencing by amplification of the 16S rDNA V3-V4 hypervariable regions followed by bioinformatic analysis. The production of SCFA in cultures was determined by GC analysis. RESULTS: Only the caecal and distal colon microbiota was able to hydrolyze and metabolize ellagitannins present in LSH to urolithins. Urolithin M6, M7, urolithin C, A and iso-urolithin A were detected together with a previously not described metabolite originating from the flavogalloyl moiety of C-glucosylic ellagitannins. LSH had no significant influence on microbiota diversity and metabolic activity, but was able to modulate its composition by significant decrease in Collinsella, Senegalimassilia, uncultured bacteria belonging to Porphyromonadaceae, Rikenellaceae RC9 gut group, Mogibacterium, Dorea, Lachnoclostridium, Lachnospiraceae UCG-004 group, Moryella, [Eubacterium] coprostanoligenes, Intestinimonas, Ruminococcaceae UCG-005, uncultured bacteria belonging to Ruminococcaceae, Acidaminococcus and Allisonella, while the relative abundance of Prevotella, Agathobacter, [Eubacterium] hallii group, Lachnospiraceae NK3A20 group, [Ruminococcus] torques group, Catenibacterium, Catenisphaera and Megasphaera increased. Significant correlations between taxa abundance and production of urolithins were determined. CONCLUSIONS: In the present study we have shown, that Lythrum salicaria herb fulfills the criteria of a potential candidate for antidiarrheal agent, which could be applied as therapy or prevention of post-weaning diarrhea in piglets. It not only modulates the gut microbiota composition without causing the dysbiosis and impairing metabolic activity, but is also a source of postbiotic metabolites, namely urolithins, which anti-inflammatory properties can be beneficial for gut health of piglets during the weaning period.

A Network Pharmacology Study on the Mechanisms of the Herbal Extract, Christina Loosestrife, for the Treatment of Nephrolithiasis.

BACKGROUND This study aimed to undertake a network pharmacology analysis to identify the active compounds of the herbal extract Christina Loosestrife, or Lysimachia Christinae (Jin Qian Cao), in the treatment of nephrolithiasis. MATERIAL AND METHODS The active components of Christina Loosestrife were identified from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and analysis platform and the online Taiwan TCM database. The potentially active compounds were screened based on their parenteral bioavailability identified from the TCMSP database. The PharmMapper integrated pharmacophore matching platform was used for target identification of active compounds in nephrolithiasis. The identified active compounds were validated by molecular docking using the systemsDock network pharmacology website. Biological functions and pathway outcomes of effective targets were analyzed using the Metascape gene annotation resource. The results were used to construct the pharmacological networks, which were visualized and integrated using Cytoscape software. RESULTS There were 16 active compounds of Christina Loosestrife and 11 nephrolithiasis-associated targets that were obtained. Functional enrichment analysis showed that Christina Loosestrife might exert its therapeutic effects by regulating pathways that included purine salvage, interleukin-4 (IL-4) and IL-13 signaling, and neutrophil degranulation. CONCLUSIONS Network pharmacology analysis of the herbal extract, Christina Loosestrife, identified multiple active compounds, targets, and pathways involved in the effects on nephrolithiasis.

Simultaneous green synthesis and in-situ impregnation of silver nanoparticles into organic nanofibers by Lythrum salicaria extract: Morphological, thermal, antimicrobial and release properties.

This research has revealed the promising, green and one-pot approach for fabrication of antimicrobial nanohybrids based on organic nanofibers including cellulose (CNF), chitosan (CHNF), and lignocellulose (LCNF) nanofibers impregnated with silver nanoparticles (AgNPs). Lythrum salicaria extract was used as a reducing agent as well as a capping agent. Formation of the spherical AgNPs ranging between 45 and 65 nm was proved by UV-Vis spectroscopy, transmission electron microscopy (TEM), and dynamic light scattering (DLS). Biomaterials supported AgNPs were characterized and compared for their morphological, thermal, release, and antimicrobial properties. The considerable influence of the phenolic compounds of L.salicaria extract on the synthesis and uniform distribution of AgNPs on nanofibers was confirmed by field emission electron microscopy (FE-SEM). Energy dispersive X-ray spectroscopy (EDX) and ICP-OES analysis of nanohybrids, reflected a high loading capacity for LCNF and also CHNF in contrast to CNF. The release of AgNPs from LCNF substrate was lower than other nanofibers but the order of antimicrobial activity of nanohybrids against E.coli and S.aureus was as this: CHNF ˃ LCNF ˃ CNF. Generally, this research suggested that the efficiency of CHNF and LCNF as immobilizing support of AgNPs is higher than CNF and L.salicaria extract was proposed as a high potential reducing and capping agent.

Differences in Metabolism of Ellagitannins by Human Gut Microbiota ex Vivo Cultures.

Ellagitannin-rich plant materials are used as popular remedies in the treatment of various inflammatory diseases. Urolithins are gut microbiota metabolites of ellagitannins and are considered responsible for in vivo health effects. Various natural products have been studied that are known sources of urolithins. However, few studies have focused on the metabolism of ellagitannin molecules. The aim of the study was to examine the metabolic fate of select ellagitannins using ex vivo cultures of human gut microbiota. Fifteen monomeric and dimeric ellagitannins, 1-O-galloyl-4,6-(S)-HHDP-ß-d-glucose (2), pedunculagin (3), potentillin (4), casuarictin (5), coriariin B (6), vescalagin (7), castalagin (8), stachyurin (9), casuarinin (10), stenophyllinin A (11), stenophyllanin A (12), salicarinin A (13), gemin A (14), agrimoniin (15), and oenothein B (16), and ellagic acid (1) were studied. The formation of the metabolites in ex vivo human microbiota cultures was monitored using UHPLC-DAD-MS/MS. Ellagitannins possessing hexahydroxydiphenoyl moieties were metabolized to 6H-dibenzo[b,d]pyran-6-one derivatives, i.e., urolithins. The observed differences in amounts of produced urolithins indicated that the individual microbiota composition and type of ingested ellagitannins could determine the rate of urolithin production. When the oral ingestion of natural products containing ellagitannins with hexahydroxydiphenoyl groups is considered, the formation of urolithins and their bioactivity should be addressed.

Genotoxic effect of Lythrum salicaria extract determined by the mussel micronucleus test.

A wide range of aquatic plants have been proven to release allelochemicals, of them phenolics and tannin are considered rather widely distributed. Tannins, however, have been demonstrated to have genotoxic capacity. In our study genotoxic potential of Lythrum salicaria L. (Purple Loosestrife, family Lythraceae) was assessed by the mussel micronucleus test, using Unio pictorum. In parallel, total and hydrolysable tannin contents were determined. Results clearly show that the extract had a high hydrolysable tannin content and significant mutagenic effect. As L. salicaria has been long used in traditional medicine for chronic diarrhoea, dysentery, leucorrhoea and blood-spitting, genotoxic potential of the plant should be evaluated not only with regard to potential effects in the aquatic ecosystem, but also assessing its safe use as a medicinal herb.

Lythrum salicaria L.-Underestimated medicinal plant from European traditional medicine. A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Purple loosestrife-Lythrum salicaria L. is a herbaceous perennial plant belonging to the Lythraceae family. It has been used for centuries in European traditional medicine. Despite Lythri herba being a pharmacopoeial plant material (Ph. Eur.), L. salicaria popularity as a medicinal plant has recently declined. The aim of the paper is to recall a traditional and historical use of L. salicaria and juxtapose it with comprehensive view on the current knowledge about its chemical composition and documented biological activities in order to bring back the interest into this valuable plant and indicate reasonable directions of future research and possible applications. MATERIALS AND METHODS: Systematic survey of historical and ethnopharmacological literature was carried out using sources of European and American libraries. Pharmacological and phytochemical literature research was performed using Scopus, PubMed, Web of Science and Reaxys databases. RESULTS: The review of historical sources from ancient times till 20th century revealed an outstanding position of L. salicaria in traditional medicine. The main applications indicated were gastrointestinal tract ailments (mainly dysentery and diarrhea) as well as different skin and mucosa affections. The current phytochemical studies have shown that polyphenols (C-glucosidic ellagitannins and C-glucosidic flavonoids) as well as heteropolysaccharides are dominating constituents, which probably determine the observed pharmacological effects. The extracts and some isolated compounds were shown to possess antidiarrheal, antimicrobial, anti-oxidant, anti-inflammatory and anti-diabetic activities. CONCLUSIONS: The intrinsic literature overview conclusively demonstrates that L. salicaria L. used to be considered as an exceptionally effective remedy in European traditional medicine. Despite its unquestionable important position from unknown reasons its popularity has been weakened during the past few decades. Unfortunately the contemporary pharmacological research is still insufficient to support its thoroughly described traditional uses. The necessity of complex studies regarding modes of action, which would directly refer to L. salicaria main traditional applications-gastrointestinal tract ailments, is strongly underlined.

Contribution of C-glucosidic ellagitannins to Lythrum salicaria L. influence on pro-inflammatory functions of human neutrophils.

The herb Lythrum salicaria L. (Lythraceae) is used in traditional medicine to treat diseases with an inflammatory background, such as haemorrhoidal disease, dysentery, chronic intestinal catarrh, eczema, varicose veins, periodontosis and gingivitis. Because these diseases are closely associated with an excessive inflammatory response of stimulated neutrophils, the influence of aqueous extract and isolated C-glucosidic ellagitannins (dimeric salicarinins A, B and C, vescalagin, castalagin) on their pro-inflammatory functions was examined. Lythrum salicaria aqueous extract was shown to modulate lipopolysaccharide (LPS)-triggered production of IL-8 (at 20 µg/mL, 16.6 ± 4.2 % inhibition) but had no influence on MMP-9 production. It was active towards cytochalasin A/f-MLP- stimulated elastase release (at 20 µg/mL, 21.5 ± 3.9 % inhibition), myeloperoxidase release (at 1 µg/mL, 26.5 ± 5.4 % inhibition) and f-MLP- and PMA-induced reactive oxygen species production (at 20 µg/mL, 67.0 ± 3.9 and 66.5 ± 1.9 % inhibition, respectively). The extract was also shown to inhibit expression of integrin CD11b on the neutrophil surface without influencing selectin CD62L shedding. Dose-dependent inhibition of hyaluronidase activity was observed with IC50 = 10.1 ± 1.2 µg/mL. The main C-glucosidic ellagitannins were shown to be responsible for all these activities with more significant participation attributable to dimeric salicarinins A, B, C. This study has demonstrated potent activity of aqueous extract on stimulated neutrophils; this enhanced response is known to cause pathological changes in skin and mucosa tissues. These observations support and explain the traditional use of the herb Lythrum salicaria to treat certain diseases with an inflammatory background. C-glucosidic ellagitannins, especially dimeric salicarinins, are the factors responsible for these effects.

Determination of C-glucosidic ellagitannins in Lythri salicariaeherba by ultra-high performance liquid chromatography coupled with charged aerosol detector: method development and validation.

INTRODUCTION: Lythri salicariaeherba is a pharmacopoeial plant material used by patients in the form of infusions in the treatment of acute diarrhoea. According to its pharmacopoeial monograph it is standardised for total tannin content, which should be not less than 5.0% using pyrogallol as a standard. Previous studies have shown that aqueous extracts from Lythri herba contain mainly ellagitannins among which vescalagin, castalagin and salicarinins A and B are dominating constituents. OBJECTIVE: To develop and validate an efficient UHPLC coupled with a charged aerosol detector (CAD) method for quantification of four major ellagitannins in Lythri salicariaeherba and in one commercial preparation. METHODS: Extraction conditions of ellagitannins from plant material were optimised. The relative response factors for vescalagin, castalagin and salicarinins A and B using gallic acid as an external standard were determined for the CAD detector. Then, a UHPLC method for quantification of ellagitannins was developed and validated. RESULTS: Four major ellagitannins were quantified in four samples of Lythri herba and in one commercial preparation. The sum of ellagitannins for each sample was determined, which varied from 30.66 to 48.80 mg/g of raw material and 16.57 mg per capsule for the preparation investigated. CONCLUSION: The first validated UHPLC/CAD UHPLC-CAD method for quantification of four major ellagitannins was developed. The universality of the CAD response was evaluated and it is shown that although all compounds analysed have similar structures their CAD response differs significantly.

Phytochemical evaluation of Lythrum salicaria extracts and their effects on guinea-pig ileum.

n-Hexane, chloroform, ethyl acetate and 50% ethanol in water extracts prepared from the air-dried flowering parts of Lythrum salicaria L. were tested for in vitro pharmacological properties on Guinea-pig ileum, which is suitable for detecting a whole range of neuronal and smooth muscle effects. UHPLC-MS was used to evaluate polyphenol components of the extracts. In the ileum, the most prominent response (46.4% related to 0.5 microM histamine) of the extracts causing smooth muscle contractions were triggered by the 50% ethanol in water extract in a concentration-dependent manner. Atropine, indomethacin and PPADS plus suramin significantly reduced the contractile response caused by this extract. The strongest inhibition was due to atropine. The results suggest that L. salicaria extracts have a moderate muscarinic receptor agonist effect in Guinea-pig ileum and that prostanoids and purinoceptor mechanisms are involved to some extent. Therefore diluted extracts of L. salicaria p.o. could be used as a mild stimulant of gastrointestinal motility. The 50% ethanol in water extract was rich in polyphenols. n-Hexane, chloroform and ethyl acetate extracts failed to contain catechin, caffeic acid, quercetin-3-D-galactoside and rutin, but they all showed spasmogenic effects, and, therefore we do not think that these compounds could be involved in the spasmogenic activity.

Biological activity and microscopic characterization of Lythrum salicaria L.

BACKGROUND: There are several plants have been used worldwide in the folk medicine with high incidence for treatment of human disorders, of which Lythrum salicaria belongs to the Lythraceae family has traditionally reputation for some medicinal usage and recently many biological and pharmacological activity of the plant have been studied. METHODS: In this study, microscopic characterizations of the aerial parts of the plant were determined. Moreover, the plant extract (aqueous methanol 80%) was subjected to an anti-diabetic activity test (in a rat model of streptozocin induced diabetes), anti-Helicobacter pylori (using disc diffusion method) along with antioxidant activity against DPPH (stable free radical) tests. Besides, total flavonoids, phenols, tannins, as well as polysaccharides contents have been assessed using spectroscopic methods. RESULTS: The microscopic properties of the plant fragments revealed anomocytic stomata, conical shape trichomes, and abundant spherical pollen grains as a characteristic pattern for the aerial parts of the plant. The extract of the plant at concentration of 15 g/kg showed mild lowering activity on blood glucose level to 12.6% and 7.3% after 2 and 3 h of administration. Additionally, clinically isolated H. pylori strain was inhibited with the plant extract at concentration of 500 mg/mL (zone of inhibition: 17 ± 0.08 mm). Moreover, IC50 values for DPPH inhibition of the plant extract, vitamin E, BHA were examined as 13.5, 14.2, and 7.8 µg/mL, respectively. Total flavonoids, phenols, tannin, and polysaccharides contents of the extract were successfully evaluated as 5.8 ± 0.4 µg QE/mg EXT, 331 ± 3.7 µg GAE/mg EXT, 340 ± 2.3 µg TAE/mg EXT, 21 ± 0.2 µg GE/mg EXT, respectively. CONCLUSIONS: The results suggested that L. salicaria has low anti-diabetic and anti-Helicobacter pylori effects, but high antioxidant activity, just the same as positive standard (vitamin E), which might be attributed to the high content of phenolic compounds in the extract.

C-glucosidic ellagitannins from Lythri herba (European Pharmacopoeia): chromatographic profile and structure determination.

INTRODUCTION: Lythri herba, a pharmacopoeial plant material (European Pharmacopoea), is obtained from flowering parts of purple loosestrife (Lythrum salicaria L.). Although extracts from this plant material have been proven to possess some interesting biological activities and its pharmacopoeial standardisation is based on total tannin content determination, the phytochemical characterisation of this main group of compounds has not yet been fully conducted. OBJECTIVE: To isolate ellagitannins from Lythri herba, determine their structures and develop chromatographic methods for their qualitative analysis. RESULTS: Five C-glucosidic ellagitannins - monomeric- vescalagin and castalagin together with new dimeric structures - salicarinins A-C, composed of vescalagin and stachyurin, vescalagin and casuarinin, castalagin and casuarinin units connected via formation of valoneoyl group, were isolated using column chromatography and preparative HPLC. Structures were determined according to (1) H and (13) C-NMR (one- and two-dimensional), electrospray ionisation-time of flight (ESI-TOF), electrospray ionisation-ion trap (ESI-MS(n) ) and circular dichroism (CD) spectra, together with acidic hydrolysis products analysis. HPTLC on RP-18 modified plates and HPLC-DAD-MS(n) on RP-18 column methods were developed for separation of the five main ellagitannins.

Chemical constituents and cytotoxic effect of the main compounds of Lythrum salicaria L.

Lythrum salicaria L. (Lythraceae), a herbaceous plant growing widely in Iran, has been well known for many centuries for its astringent and styptic properties. A phytochemical investigation of this plant, based on spectroscopic analysis, identified fourteen compounds: 5-hydroxypyrrolidin-2-one (1), umbelliferone-6-carboxylic acid (2), 3,3',4'-tri-O-methylellagic acid-4-O-beta-D-(2"-acetyl)-glucopyranoside (3), 3,3',4'-tri-O-methylellagic acid-4-O-beta-D-glucopyranoside (4), daucosterol (5), phytol (6), dodecanoic acid (7), oleanolic acid (8), 3,3',4'-tri-O-methylellagic acid (9), corosolic acid (10), beta-sitosterol (11), peucedanin (12), buntansin (13), and erythrodiol (14). All compounds, except for 8 and 11, have been isolated from L. salicaria for the first time. Cytotoxic activities of the compounds were examined against three cancerous cell lines, colon carcinoma (HT-29), leukemia (K-562), and breast ductal carcinoma (T47D), and Swiss mouse embryo fibroblast (NIH-3T3) cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and methotrexate as positive control. Compounds 5, 10, 11, and 14 were the most active against the HT-29 cell line with IC50 values of 192.7, 36.8, 38.2, and 12.8 microg/mL, respectively. Compounds 14, 11, 5, and 10 were 6.4, 2.8, 2.6, and 1.4 times, respectively, more selective than methotrexate. Compound 5 was the most active against the K-562 cell line (IC50 = 50.2 microg/mL), with a selectivity exceeding that of methotrexate 13.3 times. The results of the cytotoxic assay confirmed that growth and proliferation of the cancer cell lines are predominantly influenced by triterpene derivatives and sterols of this plant.

Antitussive and bronchodilatory effects of Lythrum salicaria polysaccharide-polyphenolic conjugate.

A high molecular mass polysaccharide-polyphenolic conjugate has been isolated from flowering parts of Lythrum salicaria by hot alkaline extraction. Its chemical analysis revealed 74% of carbohydrates and 17% of phenolics. Compositional analysis of carbohydrate part showed a high GalA content (49%), Rha (25%), Gal (13%) and Ara (9%) residues, and indicated thus rhamnogalacturonan associated with arabinogalactan in Lythrum conjugate. Antitussive activity tests, performed in three doses of Lythrum conjugate - 25, 50 and 75 mg/kg of animal body weight, showed the reduction of the number of cough efforts even 5h after administration. However, their antitussive effects were lower in comparison with that of codeine, the strongest narcotic antitussive agent. The tests evaluating the influence of different doses on airways smooth muscle reactivity revealed more significant effect of Lythrum conjugate in comparison with that of salbutamol, a commercial bronchodilator used in a clinical practice. Measurements of specific airway resistance pointed at both, the dose-dependent bronchodilatory activity and possible participation of bronchodilation on antitussive effect of Lythrum conjugate. This study represents the first sight into pharmacodynamic properties of Lythrum polysaccharide-polyphenolic glycoconjugate.

Variability of total flavonoid, polyphenol and tannin contents in some Lythrum salicaria populations.

Lythrum salicaria L. can vary in some morphological and phytochemical features according to its habitat. The present study investigates and compares the total flavonoid, polyphenol and tannin composition of twelve populations of L. salicaria collected from different ecological habitats in south-west Hungary. We studied the plant heights and soil moistures of the habitats analyzing the potential correlation between the morphological and environmental factors and chemical compositions. Total flavonoid, polyphenol and tannin contents were determined according to the valid spectroscopic methods of the European Pharmacopoeia. Total flavonoid content was higher in the populations collected during the main blooming period in August than at the beginning of flowering in July. Higher values for total polyphenol and tannin were detected in flowering branch tips in August than in July, and higher ones in leaves and shoots in July than in August. The highest flavonoid content was measured in the leaves, followed by the flowering branches and shoots as opposed to the total polyphenol and tannin contents, which were higher in the flowering branch tips than in the other organs.

Antioxidant, anti-inflammatory, anti-nociceptive activities and composition of Lythrum salicaria L. extracts.

Lythrum salicaria (purple loosestrife) known as "Tibbi hevhulma" in Turkish is used for its several beneficial health effects against as diarrhea, chronic intestinal catarrh, hemorrhoid and eczema in the form of a decoction or a fluid extract and to treat varicose veins, bleeding of the gums, hemorrhoid and eczema, externally. Dried herbal parts of Lythrum salicaria L. (Lythraceae) were sequentially extracted with different solvents such as petroleum ether, ethyl acetate, methanol and 50% aqueous methanol. Water extract of Lythrum salicaria was also prepared under reflux. Antioxidant, anti-inflammatory and anti-nociceptive activities of all the extracts were investigated using in vitro and in vivo methods, respectively. Free radical scavenging activity (1,1-diphenyl-2-picrylhydrazyl, DPPH* assay), iron(III) reductive activity, capacity of the inhibition of linoleic acid peroxidation and MDA formation, anti-nociceptive activity (p-benzoquinone-induced abdominal constriction test) and anti-inflammatory activity (carrageenan-induced hind paw edema model) were used for all the extracts. In addition, the content of total phenolics, flavonoids and flavonols in all the extracts were determined with spectrophotometric methods. Results were compared with reference antioxidants via ascorbic acid, butylated hydroxytoluene, and gallic acid. Qualitative and quantitative compositions of all the extracts were analysed using a HPLC-PDA system. Polar fractions were found to be rich in flavonoids such as isovitexin and isoorientin.

Susceptibility of the leaf-eating beetle, Galerucella calmariensis, a biological control agent for purple loosestrife (Lythrum salcaria), to three mosquito control larvicides.

We evaluated the susceptibility of Galerucella calmariensis, a species used to control purple loosestrife (Lythrum salicaria), to three mosquito control larvicides. Larvae and adults were fed loosestrife cuttings dipped in Abate (< or = 375 g x L(-1)), Altosid (< or = 250 g x L(-1)), and Bacillus thuringiensis var israeliensis (Bti) (< or = 110 g x L(-1)). Eggs on cuttings were dipped in the same concentrations. Pupae were immersed in Abate and Altosid solutions (< or = 474.4 microg x L(-1) and < or = 1,169.2 microg x L(-1), respectively). Hatching success of eggs dipped in Abate (> or = 3.75 g x L(-1)) was reduced significantly and survival was significantly lower among larvae and adults eating cuttings dipped in Abate (> or = 0.17 g x L(-1) and > or = 2.27 g x L(-1), respectively). Hatching success of eggs dipped in Altosid (> or = 2.52 g x L(-1)) was reduced significantly. With exposure to Altosid, larval survival to pupation and adult emergence was reduced significantly at concentrations of > or = 2.92 g x L(-1) and > or = 0.63 g x L(-1), respectively. Altosid (> or = 0.23 g x L(-1)) also delayed the onset of pupation and adult emergence among larvae that survived to pupate. Larvae that survived with exposure to Altosid (> or = 1.72 g x L(-1)) grew to 70% larger than those exposed to lower concentrations. Pupal survival was unaffected with exposure to Abate and Altosid and adult survival was unaffected with exposure to Altosid. Bacillus thuringiensis var israeliensis did not adversely affect any life stage of G. calmariensis. The mean Abate concentration on cuttings exposed to operational spraying was in the range that reduced egg hatchability and adult survival but was higher than concentrations that caused complete mortality of larvae. The mean Altosid concentration on cuttings exposed to operational spraying was in the range that reduced hatching success in eggs and delayed pupation and adult emergence of larvae.