Crude Organic Extracts Research Articles

Antimicrobial Fatty Acids from Green Alga Ulva rigida (Chlorophyta).

This study deals with the antimicrobial potential assessment of Ulva rigida, in regard to collection period and sampling site. Besides, we assess the chemical composition of bioactive compounds. For this purpose, Ulva rigida was seasonally collected from two northern sites in Tunisia, Cap Zebib rocky shore (CZ) and Ghar El Melh lagoon (GEM). Crude organic extracts were prepared using dichloromethane and dichloromethane/methanol and tested against 19 indicator microorganisms using the disk diffusion method and microdilution technique to determine the minimum inhibitory concentration (MIC). Silica gel column and thin layer chromatography were used for purification of active compounds. Nuclear magnetic resonance (NMR) and gas chromatography were used for compounds identification. Samples of Ulva rigida collected from the two sites have uniform antimicrobial activity throughout the year. Algae collected from the lagoon showed the largest spectrum of activity and were used for subsequent analysis. Bioguided purification of extracts from Ulva rigida, collected at GEM, leads to 16 active fractions with antibacterial effect mainly against Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212. These fractions were identified as fatty acids, mainly oleic (C18: 1 w9), linoleic (C18: 2 w6), palmitic (C16: 0), and stearic (C14: 0). MICs values ranged from 10 to 250 μg/ml.

Assessment of the antifungal potential of selected desert plant extracts against pathogenic human fungi

THE ANTIFUNGAL activities of six selected desert plants belonging to families Chenopodiaceae, Solanaceae, Brassicaceae, Fabiaceae, Zygophyllaceae and Aizoaceae from the North Western Mediterranean coastal region, Egypt, were investigated against six human pathogenic fungal species (Candida albicans, Candida tropicalis, Trichosporon sp., Aspergillus fumigatus, Aspergillus flavus and Aspergillus versicolor). Aqueous as well as organic crude extracts of the selected desert plants were screened against the different human pathogenic fungal species. Results demonstrate that the non-polar fraction of Atriplex halimus L. and Mesembryanthemum crystallinum L. exhibited the most antagonistic activity. The MIC values of fractions against yeasts and moulds ranged from 0.195 to 6.25mg/ml, whereas the fungicidal activity ranged from 0.781–12.5mg/ml. The most efficient antifungal activity was displayed by the petroleum ether fraction of M. crystallinum L. which inhibited the growth of yeast at MIC value of 0.195mg/ml and moulds at MIC values that ranged from 1.56–3.12mg/ml. Notably, the majority of combinations between plant extracts and antifungal drugs and/or plant extracts showed synergistic antifungal activities against the tested fungal species. As for the probable mechanism for the observed antifungal activity of the petroleum ether fraction of M. crystallinum, a considerable reduction in the ergosterol content and leakage of plasma and cellular membranes of the tested fungal species.

FLORAL EXTRACTS OF ALLAMANDA BLANCHETII AND ALLAMANDA CATHARTICA ARE COMPARATIVELY HIGHER RESOURCE OF ANTI-OXIDANTS AND POLYSACCHARIDES THAN LEAF AND STEM EXTRACTS

Objective: The present study undertakes a comparative analysis of the level of secondary metabolites present in the leaf, flower and stem of the two ornamental plants, Allamanda blanchetii and Allamanda cathartica.Methods: The two plant species, Allamanda blanchetii and Allamanda cathartica were collected, washed, shade dried in room temperature and powered in mechanical grinder. Phytochemicals were extracted from the power with methanol and double distilled water. The estimation of flavonoids, polyphenols, polysaccharide were done by standard methods and the anti-oxidant activity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) discoloration assay.Results: Our study reveals that the flower of both species contain highest amount of secondary metabolites in crude methanolic and aqueous extracts. In case of leaf, the methanolic extracts contain higher amount of polyphenol, flavonoid and anti-oxidant property in comparison to aqueous extracts, where as the aqueous extract contain higher amount of polysaccharide content than its counterpart. In stem, crude organic extract has higher amount of polyphenol and flavonoid and the aqueous extract has higher amount of polysaccharide and anti-oxidant property.Conclusion: The flower of Allamanda cathartica and Allamanda blanchetii has higher amount of flavonoids, polyphenols, polysaccharide and the floral extracts display comparatively higher anti-oxidant property.

Use of herbal extract from Artemisia herba-alba (Shih) in pharmaceutical preparations for dental hygiene

Antibiotic resistance in bacterial species is opening new avenues to search for alternative modes of antimicrobial treatment, medicinal plant extracts being one among them. The aim of this study was to access the possibility of medicinal plant extract from Shih in the manufacture of pharmaceutical preparations for oral hygiene specifically for the prevention and treatment of dental caries due to Streptococcus mutans. Antimicrobial effects of crude organic extract of Shih on S. mutans isolated from the saliva were examined by taking S. mutans with culture media only (−ve control); S. mutans treated with the antibiotic gentamicin (+ve control) and S. mutans treated with Shih. Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were Determination by Iodonitrotetrazolium chloride (INT) colorimetric assay Time-kill dynamic assay was performed using broth microdilution method. The metabolic reason behind the bacteriostatic and bactericidal effect were studied by measuring the glucose utilization by the microbes, pH as a measure of acid production, nucleic acids quantitation to check the DNA status and inhibition of water-insoluble glucan synthesis were undertaken. Shih MIC for S. mutans was at 2.5 mg/ml and MBC was 4 mg/ml. S. mutans bacterial population started reclining within 60 min of incubation with Shih at MBC. Utilization of added glucose was very high at MIC due to bacteria overcoming the stress, whereas at MBC its utilization was less. Accordingly pH also became acidic to 2.9 with MIC and 4.03 with MBC. There was a great degree of inhibition in the formation of nucleic acids indicating this crude extract interferes with DNA replication. Inhibition of glucan synthesis was to the tune of 45% as compared to control. Thus we conclude that Shih has potentially effective antibacterial activity hence it can be proposed as a potentially effective antiplaque and anticariogenic agent in the form of mouth wash or gum paint. However, the cytotoxicity of the extract needs to be evaluated in in-vitro and in-vivo conditions before it is considered as a safe antiplaque and anticariogenic agent.

Antitumor activity of Chlorella sorokiniana and Scenedesmus sp. microalgae native of Nuevo León State, México.

Cancer cases result in 13% of all deaths worldwide. Unwanted side effects in patients under conventional treatments have led to the search for beneficial alternative therapies. Microalgae synthesize compounds with known in vitro and in vivo biological activity against different tumor cell lines. Therefore, native microalgae from the State of Nuevo Leon, Mexico may become a potential source of antitumor agents. The aim of the present study was to evaluate the in vitro cytotoxic effect of Nuevo Leon regional Chlorella sorokiniana (Chlorellales: Chlorellaceae) and Scenedesmus sp. (Chlorococcales: Scenedesmaceae). Native microalgae crude organic extracts cytotoxicity against murine L5178Y-R lymphoma cell line and normal lymphocyte proliferation were evaluated using the MTT reduction colorimetric assay. Cell death pathway was analyzed by acridine orange and ethidium bromide staining, DNA degradation in 2% agarose gel electrophoresis and caspases activity. Results indicated significant (p < 0.05) 61.89% ± 3.26% and 74.77% ± 1.84% tumor cytotoxicity by C. sorokiniana and Scenedesmus sp. methanol extracts, respectively, at 500 µg/mL, by the mechanism of apoptosis. This study contributes to Mexican microalgae biodiversity knowledge and their potential as antitumor agent sources.

Phytochemical screening of different organic crude extracts from the stem bark of Ziziphus spina-christi (L.)

Zizyphus spina-christi (L.) (Rhamnaceae family) is a subtropical plant known as ‘Nabq’ or ‘Sidr’ which is used for various medicinal purposes. It has used in folk medicine in pain related ailments throughout these regions. Aim of our work is to phytochemically investigate the plant’s bark in our laboratory with Chromatographic approach. The crude ethanol extract of dried bark of Z. spina-christi (L.) was fractionated by diethyl ether, chloroform, ethyl acetate and finally with n-butanol. Finally the fractions were investigated for the chemical constituents by using gas chromatography-mass spectrometry. Thirty six compounds from the four fractions of the stem bark of Z. spina-christi (L.) have been identified and recognized, especially betulin, quercetin, stigmasterol, sitosterol, ethyl oleate and phytol by means of GC-MS analyses.

Endophytic Fusarium solani: A rich source of cytotoxic and antimicrobial napthaquinone and aza-anthraquinone derivatives.

This study reports the chemical investigation and bioactivity of the secondary metabolites produced by the endophytic fungus Fusarium solani isolated from Cassia alata Linn. growing in Bangladesh. This plant was collected from conservation forest in Bangladesh and belongs to the Caesalpiniaceae family. The endophytic fungus Fusarium solani was isolated from the tissue of root of this plant. The fungal strain was identified by morphological characters and DNA sequencing. The crude organic extract of the fungal strain was proven to be active when tested for cytotoxicity against Brine Shrimp Lethality Bioassay, antimicrobial and antioxidant activity. The bioactivity guided fractionation of the ethyl acetate extract leads to the isolation of seven secondary metabolites in pure form. The structures of the isolated compounds were determined by the analysis of NMR and mass spectroscopic data. Bioassay investigation of the isolated secondary metabolites suggested aza-anthraquinones are more potent bioactive compounds as anticancer and antimicrobial agent.

Composition Chimique Et Activité Antimicrobienne Des Huiles Essentielles De Deux Espèces De Lavande : Lavandula Dentata Spp. Dentata Et Lavandula Peduncultata Spp. Pedunculata

The aim of the present work is to compare the chemical composition, antibacterial and antifungal activity of two spontaneous species of lavender "Lavandula stoechas L." and "Lavandula dentata L." in Morocco. A phytochemical screening was performed on the aerial part of invested plants. The qualitative variation was revealed in the secondary metabolites of the species studied. Subsequently, the dosage of the crude organic extracts showed that the polyphenol content was higher in the methanol fraction, with an important yield in L. dentata. Also, the essential oils of the aerial part, obtained by hydrodistillation gave higher yields in L. dentata than L. pedunculata. Finally, tests of the antibacterial and antifungal activity of the essential oils of the two species were performed on four strains of bacterial pathogens (Staphylococcus aureus "Gram+," and Escherichia coli; Klebsiella pneumoniae and Proteus mirabilis which are Gram-), and three phytopathogenic fungi : Rhizopus stolonifer, Aspergillus brasiliensis and Penicillium expansum, The results obtained showed that the essential oil of L. dentata showed a strong antibacterial activity against the studied strains, particularly against S. aureus. While the antifungal activity revealed that L. pedunculata has a remarkable antifungal power against the fungi tested in comparison with L. dentata.

GC-MS analysis of the bioactive phytoconstituents of various organic crude extracts from the seed kernels of Manilkara bidentata (balata) collected in Trinidad, W.I.

The present study reports the first phytochemical investigation of the seed kernels of Manilkara bidentata (Balata) harvested in Trinidad, W.I. Gas chromatography–mass spectrometry (GC-MS) analysis of the n-hexane, chloroform, ethyl acetate, ethanol and methanol extracts showed a total of 39 components. 2,6,10,14,18-pentamethyl-2,6,10,14,18-eicosapentaene (74.93%), 9-octadecenoic acid, (Z)- 2,3-dihydroxypropyl ester (79.98%), (Z)-ethyl oleate (92.75%), Z,E-2-methyl-3,13-octadecadien-1-ol (80.51%) and 5-(hydroxymethyl)-2-furancarboxyaldehyde (50.32%) were the major constituents identified in the n-hexane, chloroform, ethyl acetate, ethanol and methanol extracts, respectively. The extracts showed the presence of several bioactive components and provides reference data for further research of its active constituents.

Sponge symbioses between Xestospongia deweerdtae and Plakortis spp. are not motivated by shared chemical defense against predators.

The recently described epizoic sponge-sponge symbioses between Xestospongia deweerdtae and two species of Plakortis present an unusual series of sponge interactions. Sponges from the genus Plakortis are fierce allelopathic competitors, rich in cytotoxic secondary metabolites, and yet X. deweerdtae flourishes as an epizoic encrustation on Plakortis deweerdtaephila and Plakortis symbiotica. Our objective in this study was to evaluate the hypothesis that X. deweerdtae grows epizoic to these two species of Plakortis due to a shared chemical defense against predators. We collected free-living individuals of X. deweerdtae and symbiotic pairs from a wide geographical range to generate crude organic extracts and a series of polarity fractions from sponge extract. We tested the deterrency of these extracts against three common coral reef predators: the bluehead wrasse, Thalassoma bifasciatum, the Caribbean sharpnose puffer, Canthigaster rostrata, and the white spotwrist hermit crab, Pagurus criniticornis. While the chemical defenses of P. deweerdtaephila and P. symbiotica are more potent than those of X. deweerdtae, all of the sponge species we tested significantly deterred feeding in all three generalist predators. The free-living form of X. deweerdtae is mostly defended across the region, with a few exceptions. The associated form of X. deweerdtae is always defended, and both species of Plakortis are very strongly defended, with puffers refusing to consume extract-treated pellets until the extract was diluted to 1/256× concentration. Using diode-array high performance liquid chromatography (HPLC) coupled with high-resolution mass spectrometry (LC-MS/IT-TOF), we found two secondary metabolites from P. deweerdtaephila, probably the cyclic endoperoxides plakinic acid I and plakinic acid K, in low concentrations in the associated—but not the free-living—form of X. deweerdtae, suggesting a possible translocation of defensive chemicals from the basibiont to the epibiont. Comparing the immense deterrency of Plakortis spp. extracts to the extracts of X. deweerdtae gives the impression that there may be some sharing of chemical defenses: one partner in the symbiosis is clearly more defended than the other and a small amount of its defensive chemistry may translocate to the partner. However, X. deweerdtae effectively deters predators with its own defensive chemistry. Multiple lines of evidence provide no support for the shared chemical defense hypothesis. Given the diversity of other potential food resources available to predators on coral reefs, it is improbable that the evolution of these specialized sponge-sponge symbioses has been driven by predation pressure.

Phytochemical analysis of some medicinal plants of district Saharanpur

District Saharanpur is a rich source of medicinal plants because of its physical features, the north and the north-east of the district is surrounded by Shivalik hills and having rich biodiversity. The objective of present study was to investigate the presence of phytochemicals in five medicinal plants such as, Tinospora cordifolia (Thunb.)Miers, Bryophyllum pinnatum (Lam.)Oken , Terminalia belerica Roxb., Xanthium strumarium Linn. and Oldenlandia corymbosa Linn. For the study the solvents used were water, methanol, ethanol and acetone. Soxhlet apparatus was used for the organic solvent extraction. Carbohydrates, proteins, flavonoids, tannins, saponin, and phenols were detected in these selected medicinal plants. Our analysis proved that crude aqueous and organic solvent extracts of these plants having the bioactive compounds which are medicinally important for the use in the treatment of various diseases

A Sphingosine-type cerebroside in Clavicorona pyxidata induce fruit body formation

Clavicorona pyxidata is a wild edible and medicinal mushroom that is rich in bioactive natural products and has thus been extensively used as traditional medicine in China. The present study has determined that the organic crude extract prepared from a fermented culture of C. pyxidata imparted auto-inhibitory effects on mycelial growth and then induced the formation of fruiting bodies. By monitoring bioactivity, one compound was isolated via successive chromatography over silica gel, Sephadex LH-20, and Cl8-reversed phase silica gel and was identified as a known sphingosine-type cerebroside by nuclear magnetic resonance (NMR) and physicochemical data, namely, (4E, 8E)-N-D-2′-hydroxypalmitoyl-1-O-β-D-glucopyranosyl-9-methyl-4,8-sphingadienine. The application of this cerebroside at a concentration of 200 μg/disc paper resulted in the inhibition of aerial hyphal growth of C. pyxidata. The findings of the present study indicated that this C. pyxidata cerebroside is a fruiting body-inducing substance (FIS).

Latitudinal variation in the microbiome of the sponge Ircinia campana correlates with host haplotype but not anti-predatory chemical defense

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 565:53-66 (2017) - DOI: https://doi.org/10.3354/meps12015 Latitudinal variation in the microbiome of the sponge Ircinia campana correlates with host haplotype but not anti-predatory chemical defense Clark M. Marino1,2, Joseph R. Pawlik1, Susanna López-Legentil1, Patrick M. Erwin1,* 1Center for Marine Science and Department of Biology and Marine Biology, University of North Carolina Wilmington, Wilmington, NC 28409, USA 2Present address: Department of Ecology, Evolution, and Marine Biology, University of California Santa Barbara, Santa Barbara, CA 93106, USA *Corresponding author: erwinp@uncw.edu ABSTRACT: Many marine sponges host diverse symbiotic microbial communities that have been implicated in the production of secondary metabolites. These metabolites may defend the host sponge from potential predators. Variability in symbiont communities across the range of the host sponge could alter levels of chemical defense. To investigate the relationship between symbiont composition and chemical defense, the microbiomes and palatability of tissue samples from Ircinia campana were characterized from 5 sites along a latitudinal gradient spanning temperate (South Atlantic Bight, SAB) and tropical (Caribbean) regions. Terminal restriction fragment length polymorphism analysis and Illumina sequencing of 16S ribosomal RNA genes revealed that I. campana from different locations contained significantly distinct microbiomes and exhibited a consistent relationship of lower symbiont similarity over greater geographic distance (i.e. distance-decay). However, crude organic extracts of all samples of I. campana were unpalatable to assay fish Thalassoma bifasciatum in laboratory assays, indicating no difference in chemical defense across locations. Distinct haplotypes of I. campana were detected in populations from the SAB and Caribbean, correlating with the observed patterns of latitudinal variation in microbial symbiont communities. Our findings indicate that I. campana is chemically defended from fish predators across the range of the species and that latitudinal variation occurs in the microbiome of I. campana, driven by a combination of host-specific factors and region-specific environmental filtering of symbiont communities. KEY WORDS: Symbiosis · Microbial community · Chemical defense · Biogeography · Porifera · Intraspecific variation Full text in pdf format Supplement 1 Supplement 2 PreviousNextCite this article as: Marino CM, Pawlik JR, López-Legentil S, Erwin PM (2017) Latitudinal variation in the microbiome of the sponge Ircinia campana correlates with host haplotype but not anti-predatory chemical defense. Mar Ecol Prog Ser 565:53-66. https://doi.org/10.3354/meps12015 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 565. Online publication date: February 17, 2017 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2017 Inter-Research.

Antimicrobial activity, toxicity and phytochemical screening of selected medicinal plants of Losho, Narok County, Kenya

Chalo DM, Lukhoba C, Fidahussein DS, Nguta JM. 2017. Antimicrobial activity, toxicity and phytochemical screening of selected medicinal plants of Losho, Narok County, Kenya. Biofarmasi (Rumphius J Nat Prod Biochem) 15: 29-43. In Kenya, microbial infections are a major cause of morbidity. The effectiveness of antibiotic is threatened by the increase of resistance of pathogenic microbes against most available drugs because new pathogens continue to emerge. Nowadays, herbal remedies offer hope since they are readily available and cheap. The aim of this research was to investigate the activity of antimicrobial, the lethality of brine shrimp and phytochemical composition of crude extracts of four selected plants namely Schrebera alata (Oleaceae), Ormocarpum kirkii (Papilionoideae), Helichrysum forskahlii (Asteraceae) and Cussonia holstii (Araliaceae) that are medicinally used by herbalists from Losho, Narok County Kenya for treatment of ear, nose and throat infections, gastrointestinal disorders and skin diseases. Qualitative antimicrobial susceptibility test against five microorganisms, methicillin-resistant Staphylococcus aureus, Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans was investigated using agar diffusion methods to produce inhibition zones and the data accrued were analyzed using Analysis of variance. Minimum inhibitory concentrations were determined by broth microdilution method. Toxicity of the extracts was analyzed using brine shrimp lethality assay. The median fatal concentration of fifty was determined by data analysis using Finney‟s computer program. Phytochemical screening for flavonoids, sterols, alkaloids, tannins, quinones and terpenoids and saponins was determined using standard procedures. The observation showed that the organic crude extracts of H. forskahlii had the highest inhibition zone against methicillin-resistant S. aureus of 19.5 and 18.5 mm in agar well and agar disk diffusion respectively. In addition, organic extracts of H. forskahlii showed the highest antifungal inhibition zone of 8.5 mm in agar well diffusion. Minimum values of inhibitory concentrations varied from 15.625 to 250 mg/mL. Organic crude extracts of H. forskahlii and C. holstii were found to be highly toxic with lethal concentration of 0.009 mg/mL. All plant crude extracts contained flavonoids, sterols, alkaloids, tannins, quinones, and terpenoids. Saponins were present in all the plant extracts except in organic extracts of H. forskahlii. This study promoted the first record of antimicrobial activity, toxicity and phytochemical composition of S. alata and C. holstii. The study has shown that H. forskahlii and O. kirkii possess promising antimicrobial activity against microbes of health importance and could lead to the isolation of new, safe and efficacious antimicrobial compounds. Further research should be carried on O. kirkii and S. alata to isolate and characterize the compounds responsible for the observed activity.

&amp;lt;i&amp;gt;In Vitro&amp;lt;/i&amp;gt; Antibacterial, Antifungal and Other Medical Properties of Endangered Medicinal Plant Seeds

The risk created by infectious microorganisms to humans attracted the development of common medicine. To find an alternative source, medicinal plants with diverse metabolites play an important role in curing the diseases and human disorders caused by microbial pathogens. Medicinal plants namely, Citrullus colocynthis, Hyoscyamus muticus, Ocimum basilicum, Amaranthus lividus, Salvia aegyptiaca and Ruta chalepensis are commonly used as a traditional medicine in Gulf countries. The present study aimed to investigate the antibacterial, antifungal and antioxidant potential of the organic crude extracts obtained from the seeds. Besides, the possible antimicrobial mechanisms of the extracts were evaluated by determining the enzyme activities. The antibacterial and antifungal activities of the crude extracts were evaluated by the broth micro dilution method and the effect of the extracts on the pathogens were determined by quantifying the alkaline phosphatase (ALP), lactate dehydrogenase (LDH) enzymes and intracellular protein leakage. Besides, the antioxidant properties were determined using hydroxyl radical scavenging assay, DPPH radical scavenging assay, reducing power assay and superoxide radical scavenging assay. Results indicated that the extracts of C. colocynthis showed promising activity against all the tested pathogens, especially the MIC values were ranged from 100 to 150 μg/ml for Gram positive bacteria and 100 to 250 μg/ml for Gram negative bacteria respectively. The MIC values of H. muticus, O. basilicum and R. chalepensis against the fungal pathogens were ranged from 100 to 500 μg/mL respectively. The ALP activity was higher in extract treated Klebsiella pneumoniae compared with control, whereas the LDH and protein concentrations for Escherichia coli and Staphylococcus aureus were comparatively higher. Furthermore, all the studied seed extract showed good antioxidant activities. In conclusion, the studied plant seed extracts documented good antimicrobial and antioxidant activities. Therefore, the medicinal plants would be the excellent source for natural antioxidant and antibacterial agents for medical and applications.

Effects similar to anxiolysis in an organic extract of Stypopodium zonale on an anxiety-related behavior in Drosophila melanogaster

Nature has a great diversity of organisms whose bioactive compounds may potentially be studied. When it comes to aquatic life we find that algae are organisms that are well suited for screening and identification of bioactive compounds due to their widespread distribution in both salt and freshwater. Our hypothesis is that a crude organic extract of the brown algae Stypopodium zonale can decrease anxiety-related behaviors in Drosophila melanogaster. Stypopodium zonale was collected in the south coast of Puerto Rico and the potential anxiolytic-like effects of the extract were studied in an anxiogenic-like behavioral paradigm in Drosophila melanogaster. This behavior is called centrophobia and is measured using an Open Field Arena (OFA). Validation of the paradigm gave the expected results as reported in the literature, in which Drosophila exhibits a phobia (avoidance) of remaining in the center of the OFA, which corresponds to a behavior with anxiety components. The organic extract was dissolved with dimethyl sulfoxide (DMSO). Toxicity tests were performed both for DMSO and the crude organic extract, and neither showed positive results. To perform the behavioral trials, 1 mL of the crude extract and 4 mL of water were mixed with 1.8 g of Drosophila food. The final concentration of the crude extract in the food was 5.4 mg/mL. The adult flies were grown in a tube with the extract until a considerable quantity of larvae was observed, and then the adults were removed. These new larvae, once turned into adult flies, were used for the behavioral trials. The behavior of control flies (food without extract) and experimental flies (extract containing food) was recorded with a video camera and the results of the centrophobic behavior were analyzed and compared using quantitative criteria. Both the control and experimental trials were performed in triplicate. The results show that flies grown in food containing the crude extract present a significant reduction in centrophobia compared with control flies. In conclusion, our results suggest that the organic crude extract from Stypopodium zonale has anxiolytic-like effects in a Drosophila melanogaster model with anxiety components. We are currently performing Nuclear Magnetic Resonance (NMR) studies on the crude extracts to identify the most abundant secondary metabolites. Future experiments should include the administration of the crude extracts (or fractions of the most abundant secondary metabolites) to a vertebrate model in to test the effect in a behavior with anxiety components. We are also in the process of developing a preliminary model of possible mechanisms of action of the crude organic extract in the reduction of centrophobia. KEYWORDS: Anxiety; Algae; Drosophila melanogaster; Open Field Arena; Organic extracts; Centrophobia; Stypopodium zonale

P-Sulfooxyphenylpyruvic acid derivatives from the green macroalga Caulerpa taxifolia

The green macroalga Caulerpa taxifolia has been of a great research interest since its accidental introduction in the Mediterranean sea in 1984. This specie has yielded several structurally diverse and biologically active compounds such as caulerpenyne (Fig. 1) [1]. This metabolite, constituted by a diacetoxybutadiene moiety, exhibited very potent antibacterial properties [2]. Within our ongoing research projects devoted to the isolation of new bioactive metabolites from Mediterranean marine organisms, a specimen of the freshly collected green algae Caulerpa taxifolia, was investigated. The crude polar organic extract was subjected to a series of chromatographic separations that has led to the isolation of new metabolites (1a and 1b) structurally close to the already reported ones (2a and 2b) (Fig. 2) [3].

English

The present study is aimed to investigate phytochemical and pharmacological activities of crude organic fractional extracts of Maesa acuminata (MA). Preliminary test for phytoconstituents shows the presence of alkaloid, glycoside, steroids, tannins, saponins and reducing sugars. Fractionation was done by ethanol (leaf: ELMA), chloroform (bark: CBMA) and n-hexane (leaf: HLMA). The crude fractions were then subjected for antioxidant, anti-inflammatory and membrane stabilizing, antimicrobial and analgesic activities. In the antioxidant test performed by 1,1-dipheny-picrylhydrazyl (DPPH) assay, the CBMA and HLMA significantly scavenged DPPH radicals, where HLMA at 100 &mu;g/ml inhibited DPPH by 57.01% and the standard, ascorbic acid (AA) by 95.47%, respectively. In the disc diffusion antimicrobial sensitivity test, all the crude fractions of MA produce zones of inhibition between 8 to 18 mm. ELMA showed prominent antimicrobial activity than the CBMA and HLMA, where it strongly inhibited Staphylococcus aureus with the inhibition zones by 18 mm at 500 &mu;g/disc. Minimum inhibitory concentration (MIC) detected with the CBMA, ELMA and HLMA were between 61.25 and 250 &mu;g/ml for the test microorganisms. In the anti-diarrheal activity the HLMA and CBMA in oral dose (p.o.) showed 62.79 and 61.62% of percent inhibition of defection as compared to standard, loperamide (93.00%). Additionally, the fractional extracts of MA also possess anti-inflammatory, membrane stabilization and analgesic activities. In conclusion, M. acuminata may be a good source of therapeutic components. Key words: Antioxidant, antimicrobial, Maesa acuminata. &nbsp

Detection and Quantification of Rotenoids from Clitoria fairchildiana and its Lipids Profile

This work describes the isolation and quantification of rotenoids from crude organic extracts of different parts of Clitoria fairchildiana R. A. Howard (Leguminosae) by HPLC-DAD. The lipid composition and the Artemia salina cytotoxic activities of the isolates were also conducted. Clitoriacetal (1), 6-deoxyclitoriacetal (2), stemonal and stemonone were isolated by chromatographic procedures and identified by usual spectroscopic and spectrometric techniques. Clitoriacetal and 6-deoxyclitoriacetal were not found in all parts of the plant, such as leaves and petals, but in the roots they occur in higher concentration. The activity against brine shrimp revealed that the root extract (LD50 = 158 ppm) was the more active.

In vitro therapeutic index application in high throughput screening of the anti‐inflammatory effects of natural products evaluation in different cell lines

One thousand and four hundred crude organic extracts from natural plants, botanical chemicals were evaluated for anti‐inflammatory activity LPS treated cell lines: BV‐2 microligal cells, RAW 264.7 macrophages and C6 glioma cells. LPS is commonly used to invoke in vitro inflammation. However, studies using immortalized macrophages, microglia or astrocytoma (malignant phenotype) could result in an interfering variable when evaluating natural products which are both anti‐inflammatory and anti‐cancer (pro‐apoptotic) at similar/close concentrations. In this study, a high throuput HTP study was conducted on inflammatory parameters using an in vitro therapeutic index (iTI) = ratio of toxicity (LC50) / anti‐inflammatory potency (IC50) compared to reference controls (e.g. hydrocortisone, dexamethasone, iNOS inhibitor N6‐(1‐iminoethyl)‐L‐lysine (L‐NIL) and other OTC NSAIDs). Preliminary profiling studies showed (1) LPS activation of BV2 cells resulted &gt; 10‐fold rise in IL‐6, MIP‐2, MIP‐1g, RANTES and nitric oxide (NO); (2) LPS activated RAW 264.7 cells generated &gt; 10‐fold rise in sTNFR2, MCP‐1, IL‐6, GCSF, RANTES and NO (3) LPS/IFN‐g activated C6 glioma initiated &gt;10‐fold rise in MCP‐1, CINC1, CINC2a, CINC3 and NO. Using NO2‐ production as a guideline molecule for HTP screening, the data show that of all compounds tested : the highest anti‐inflammatory differential LC50/IC50s (Ti ) with efficacy &lt; 250μg/mL occurred in the presence of Ashwaganda, Elecampane, Feverfew, Tansy, Turmeric, Bayleaf, Rosemary, Green Tea, Yerba Santa and Centipeda Herb. Likewise, naturally derived substances with the highest anti‐inflammatory differential LC50/IC50s (Ti ) were established for cardamonin and to a lesser extent: butein, apigenin, EGCG, curcumin and quercetin. These substances had capacity to reduce NO, downregulate iNOS protein expression, significantly attenuate IL‐6 in activated RAW/BV2 cells and reduce levels of cytokine‐induced neutrophil chemoattractant CINC‐1/3, in C6 cells. These particular natural substances seem to hold therapeutic value above many others tested, with possible applications for future prevention of neuro inflammatory conditions / treatment of general inflammatory chronic disorders and possibly drug development for strategies to prevent multi‐organ damages associated with endotoxic shock.Support or Funding InformationNIMHD Grants G12MD007582 and Grant Number 1P20 MD006738