HPLC-DAD analysis and antimicrobial activities of Spondias mombin L. (Anacardiaceae).

Spondias mombin is used in the folk medicine for the treatment of diarrhea and dysentery, indicating that extracts obtained from this species may present pharmacological activities against pathogenic microorganisms. The purpose of this work was to investigate the chemical composition and evaluate the antimicrobial activity of extracts obtained from the leaves (aqueous) and bark (hydroethanolic) of S. mombin both as single treatments and in combination with conventional drugs. Following a qualitative chemical prospection, the extracts were analyzed by HPLC-DAD. The antimicrobial activities were evaluated by microdilution. The combined activity of drugs and extracts was verified by adding a subinhibitory concentration of the extract in the presence of variable drug concentrations. The Minimum Fungicidal Concentration (MFC) was determined by a subculture of the microdilution test, while the effect of the in vitro treatments on morphological transition was analyzed by subculture in moist chambers. While the qualitative analysis detected the presence of phenols and flavonoids, the HPLC analysis identified quercetin, caffeic acid, and catechin as major components in the leaf extract, whereas kaempferol and quercetin were found as major compounds in the bark extract. The extracts showed effective antibacterial activities only against the Gram-negative strains. With regard to the combined activity, the leaf extract potentiated the action of gentamicin and imipenem (against Staphylococcus aureus), while the bark extract potentiated the effect of norfloxacin (against S. aureus), imipenem (against Escherichia coli), and norfloxacin (against Pseudomonas aeruginosa). A more significant antifungal (fungistatic) effect was achieved with the bark extract (even though at high concentrations), which further enhanced the activity of fluconazole. The extracts also inhibited the emission of filaments by Candida albicans and Candida tropicalis. Together, these findings suggest that that the extract constituents may act by favoring the permeability of microbial cells to conventional drugs, as well as by affecting virulence mechanisms in Candida strains.

HPLC/DAD, Antibacterial and Antioxidant Activities of Plectranthus Species (Lamiaceae) Combined with the Chemometric Calculations.

The increase in antibiotic resistance and the emergence of new bacterial infections have intensified the research for natural products from plants with associated therapy. This study aimed to verify the antibacterial and antioxidant activity of crude extracts of the genus Plectranthus species, being the first report on the modulation of aminoglycosides antibiotic activity by Plectranthus amboinicus extracts. The chemical composition was obtained by chemical prospecting and High-Performance Liquid Chromatography with diode arrangement detector (HPLC/DAD). The antibacterial activities of the extracts alone or in association with aminoglycosides were analyzed using the microdilution test. The antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging. The phytochemical prospection allowed the flavonoids, saponins, tannins and triterpenoids to be identified. Quercetin, rutin, gallic acid, chlorogenic acid, caffeic acid, catechin, kaempferol, glycosylated kaempferol, quercitrin, and isoquercitrin were identified and quantified. The principal component analysis (PCA) observed the influence of flavonoids and phenolic acids from Plectranthus species on studied activities. Phytochemical tests with the extracts indicated, especially, the presence of flavonoids, confirmed by quantitative analysis by HPLC. The results revealed antibacterial activities, and synergistic effects combined with aminoglycosides, as well as antioxidant potential, especially for P. ornatus species, with IC50 of 32.21 µg/mL. Multivariate analyzes show that the inclusion of data from the antioxidant and antibacterial activity suggests that the antioxidant effect of these species presents a significant contribution to the synergistic effect of phytoconstituents, especially based on the flavonoid contents. The results of this study suggest the antibacterial activity of Plectranthus extracts, as well as their potential in modifying the resistance of the analyzed aminoglycosides.

Impact of drying processes on Bryophyllum pinnatum phenolic constituents and its anti-inflammatory and antioxidative activities in human erythrocytes.

The effect of drying on the phytoconstituents, antioxidative, and anti-inflammatory properties of Bryophyllum pinnatum leaves was investigated. The phenolic constituents were characterized using HPLC-DAD. The aqueous extraction was done and various assays (Inhibition of membrane stabilization, albumin Denaturation and heat-induced hemolysis, malondialdehyde (MDA), and reduced glutathione (GSH) contents, as well as superoxide dismutase (SOD) activity), were carried out on human erythrocytes. The fresh portion (89.12 µg/ml) exhibited the highest potential to inhibit heat-induced hemolysis compared to the standard drug-Diclofenac (91.51 µg/ml). Freeze-dried sample showed the highest inhibitory potential on albumin denaturation ([Freeze-dried-330.72 µg/ml], [Diclofenac-318.63 µg/ml]) and membrane destabilization ([Freeze-dried-331.93 µg/ml], [Diclofenac-289.57 µg/ml]) when compared with Diclofenac. Similarly, the freeze-dried sample showed the highest GSH and SOD level and lowest MDA level when human erythrocytes challenged with tertiary butyl hydroperoxide (tBHP) were treated with the extract. This study confirms the retention of a considerable quantity of bioactive constituents of plants when freeze-dried. PRACTICAL APPLICATIONS: The ideal method of drying Bryophyllum pinnatum and possible anti-inflammatory potential was investigated. This work may apply to the development of anti-inflammatory agents from a natural source with little or no side effect in managing inflammation.

Kava decreases the stereotyped behavior induced by amphetamine in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Kava extract (Piper methysticum) is a phytotherapic mainly used for the treatment of anxiety. Although the reported effects of Kava drinking improving psychotic symptoms of patients when it was introduced to relieve anxiety in aboriginal communities, its effects on models of psychosis-like symptoms are not investigated. AIM OF THE STUDY: To investigate the effects of Kava extract on behavioral changes induced by amphetamine (AMPH) and its possible relation with alterations in monoamine oxidase (MAO) activity. MATERIALS AND METHODS: Mice received vehicle or Kava extract by gavage and, 2 h after vehicle or AMPH intraperitoneally. Twenty-five minutes after AMPH administration, behavioral (elevated plus maze, open field, stereotyped behavior, social interaction and Y maze) and biochemical tests (MAO-A and MAO-B activity in cortex, hippocampus and striatum) were sequentially evaluated. RESULTS: Kava extract exhibited anxiolytic effects in plus maze test, increased the locomotor activity of mice in open field test and decreased MAO-A (in cortex) and MAO-B (in hippocampus) activity of mice. Kava extract prevented the effects of AMPH on stereotyped behavior and, the association between Kava/AMPH increased the number of entries into arms in Y maze test as well as MAO-B activity in striatum. However, Kava extract did not prevent hyperlocomotion induced by AMPH in open field test. The social interaction was not modified by Kava extract and/or AMPH. CONCLUSION: The results showed that Kava extract decreased the stereotyped behavior induced by AMPH at the same dose that promotes anxiolytic effects, which could be useful to minimize the psychotic symptoms in patients.

Effects of Gender and Geographical Origin on the Chemical Composition and Antiradical Activity of Baccharis myriocephala and Baccharis trimera.

This study aimed to characterize and compare essential oils and ethyl acetate fractions obtained in basic and acidic conditions from both male and female Baccharis species (Baccharis myriocephala and Baccharis trimera) from two different Brazilian regions. Samples were characterized according to their chemical compositions and antiradical activity by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. Principal component analysis (PCA) provided a clear separation regarding the chemical composition of essential oils from the samples obtained from different regions by using gas chromatography-mass spectrometry with flame-ionization detection (GC-MS-FID). PCA also revealed that gender and region of plant collections did not influence the chemical composition and antiradical activity of ethyl acetate fractions, which was corroborated with hierarchical cluster analysis (HCA) data. High performance liquid chromatography with diode-array detector (HPLC-DAD) identified significant quantities of flavonoids and phenolic acids in the fractions obtained in basic and acidic fractions, respectively. The obtained results clearly demonstrated that the geographical region of plant collection influenced the chemical composition of essential oils from the studied Baccharis species. Moreover, the obtained fractions were constituted by several antiradical compounds, which reinforced the usage of these species in folk medicine.

Investigation of the In Vitro Antioxidant Potential Of Polyphenolic-Rich Extract of Artocarpus heterophyllus Lam Stem Bark and Its Antidiabetic Activity In Streptozotocin-Induced Diabetic Rats.

Artocarpus heterophyllus Lam (Moraceae) stem bark has been used locally in managing diabetes mellitus with sparse scientific information. This study investigates the in vitro antioxidant potential of polyphenolic-rich extract of A heterophyllus stem bark as well as its antidiabetic activity in streptozotocin-induced diabetic rats. Fifty male Wistar rats were used with the induction of diabetes by a single intraperitoneal injection of streptozotocin (45 mg/kg body weight) and were orally administered 400 mg/kg free and bound phenols of A heterophyllus stem bark. The animals were sacrificed on the 28th day of the experiment using the cervical dislocation method; antihyperglycemia and anti-inflammatory parameters were subsequently assessed. The polyphenolic extracts demonstrated antioxidant potentials (such as hydrogen peroxide and diphenyl-1-picrylhydrazyl), as well as strong inhibitory activity against amylase and glucosidase. There was a significant (P < .05) increase in glycogen, insulin concentration, pancreatic ß-cell scores (HOMA-ß), antioxidant enzymes and hexokinase activities, as well as glucose transporter concentration in diabetic animals administered the extracts and metformin. Also, a significant (P < .05) reduction in fasting blood glucose, lipid peroxidation, glucose-6-phosphatase, and all anti-inflammatory parameters were observed in diabetic rats administered the extracts and metformin. The extracts demonstrated antidiabetic potential, which may be useful in the management of diabetes mellitus.

Croton campestris A. St.-Hill Methanolic Fraction in a Chlorpyrifos-Induced Toxicity Model in Drosophila melanogaster: Protective Role of Gallic Acid.

Croton campestris A. St-Hill popularly known as "velame do campo" is a native species of the savannah from northeastern Brazil, being used in folk medicine due to its beneficial effects in the treatment of many diseases, inflammation, detoxification, gastritis, and syphilis; however, its potential use as an antidote against organophosphorus compound poisoning has not yet been shown. Here, the protective effect of the methanolic fraction of C. campestris A. St.-Hill (MFCC) in Drosophila melanogaster exposed to chlorpyrifos (CP) was investigated. Flies were exposed to CP and MFCC during 48 h through the diet. Following the treatments, parameters such as mortality, locomotor behavior, and oxidative stress markers were evaluated. Exposure of flies to CP induced significant impairments in survival and locomotor performance. In parallel, increased reactive oxygen species and lipoperoxidation occurred. In addition, the activity of acetylcholinesterase was inhibited by CP, and superoxide dismutase and glutathione S-transferase activity was induced. Treatment with MFCC resulted in a blockage of all CP-induced effects, with the exception of glutathione S-transferase. Among the major compounds found in MFCC, only gallic acid (GA) showed a protective role against CP while quercetin and caffeic acid alone were ineffective. When in combination, these compounds avoided the toxicity of CP at the same level as GA. As far as we know, this is the first study reporting the protective effect of MFCC against organophosphate toxicity in vivo and highlights the biotechnological potential of this fraction attributing a major role in mediating the observed effects to GA. Therefore, MFCC may be considered a promising source for the development of new therapeutic agents for the treatment of organophosphate intoxications.

Anxiolytic and antidepressant-like effects of Annona coriacea (Mart.) and caffeic acid in mice.

This research evaluated the anxiolytic and antidepressant-like effects of a hydroethanolic extract from the leaves of Annona coriacea (EHFAC) and caffeic acid (CA). Mice were intraperitoneally treated with saline, EHFAC (1, 10, 20 mg/kg) or CA (0.15 mg/kg) and subject to the elevated plus-maze, open field, rota-rod, forced swimming and reserpine-induced akinesia tests. Pro-convulsant and anticholinergic effects were also evaluated. EHFAC presented anxiolytic-like effect on the elevated plus-maze, which was partially reversed by flumazenil. A similar effect was observed with CA. In the forced swimming test, EHFAC and CA reduced the immobility time of mice; such effect was potentiated when EHFAC or CA were associated with imipramine, bupropion and fluoxetine. The antidepressant-like effect was reinforced as EHFAC partially reversed the reserpine-induced akinesia. In addition, a pre-treatment with EHFAC and CA did not decrease the latency to 1st seizure of animals that received a sub-convulsive dose of PTZ, nor reduced the intensity of oxotremorine-induced tremors. Taken together, the results indicate that EHFAC and CA have anxiolytic and antidepressant-like effects, which involve important neurotransmitter systems, such as GABAergic and monoaminergic ones, being devoid of side effects, commonly associated with classical psychotropic drugs.

Fluted pumpkin (Telfairia occidentalis) seed modulates some markers of erectile function in isolated rat's corpus cavernosum: Influence of polyphenol and amino acid constituents.

Pumpkin seeds are often used in traditional medicine in the management of erectile dysfunction. However, there is insufficient information about the possible biochemical rationale behind this practice. Hence, this study investigated the influence of fluted pumpkin seed on critical enzymes involved in erectile function in isolated rats' corpus cavernosum in vitro. The phenolics and amino acid contents of fluted pumpkin seed were determined using HPLC-DAD and GC-PFPD analyses respectively. The aqueous extract of the fluted pumpkin seed significantly (p < .05) scavenged free radicals and inhibited PDE-5, arginase, AChE, and ACE in rats' corpus cavernosum in a concentration-dependent pattern. Quercitrin and luteolin were the most dominant phenolics, while arginine, aspartate, and cysteine were the most aboundant amino acid constituents. The positive modulatory effect of the fluted pumpkin seed on these critical markers of erectile function could be attributed to its polyphenolics and amino acid constituents. PRACTICAL APPLICATIONS: This study brought to limelight the medicinal importance of fluted pumpkin seed in erectile functions. Therefore, this seed could be used as a functional food ingredient in the management of erectile dysfunctions and also in improving erectile functions in men. In addition, the dominant phenolics and amino acid constituents of this seed might be an effective nutraceutical in enhancing erections in men.

Dietary supplementation of jute leaf (Corchorus olitorius) modulates hepatic delta-aminolevulinic acid dehydratase (δ-ALAD) activity and oxidative status in high-fat fed/low streptozotocin-induced diabetic rats.

Impaired liver function is associated with decreased hepatic delta-aminolevulinic acid dehydratase (δ-ALAD) activity in diabetes mellitus. Hence, this study described the effect of dietary jute leaf (Corchorus olitorius) on hepatic δ-ALAD activity in high-fat fed combined with low-dose streptozotocin administered diabetic rats. Animals were fed diets containing 35% fat for 14 days prior to a single administration of low-dose (35 mg/kg body weight) streptozotocin to induce diabetes. Thereafter, the animals were randomly placed in groups and fed 100 mg/g jute leaf-supplemented diets for 30 days. The result showed that jute leaf supplementation significantly (p < 0.05) reversed the decreased hepatic δ-ALAD activity, increased hepatic catalase and SOD activity accompanying the decrease in serum AST and AST activities. This finding suggests that restoration of hepatic δ-ALAD activity, modulation of hepatic function biomarkers, and increase in antioxidant status could be possible underlying events mediating the hepatoprotective effect of jute leaf in diabetic conditions. PRACTICAL APPLICATIONS: Decrease in hepatic δ-ALAD activity has been associated with diabetes-induced hepatotoxicity arising from prolonged and uncontrolled hyperglycemia. Therefore, increased δ-ALAD activity represents improved hepatic function in diabetic situations. Antidiabetic properties of jute leaf have been demonstrated but information on its effect on hepatic δ-ALAD is lacking. Thus, this study revealed that dietary supplementation of jute leaf restored hepatic δ-ALAD activities and improved liver antioxidant status in diabetic rats which is an indication of its hepatoprotective properties.

Ilex paraguariensis Attenuates Changes in Mortality, Behavioral and Biochemical Parameters Associated to Methyl Malonate or Malonate Exposure in Drosophila melanogaster.

Methylmalonic acidemia is a genetic disease characterized by accumulation of organic acids, such as methylmalonic (MMA) and malonic (MA) acids. Considering that the accumulation of MMA and MA causes several damages due to oxidative stress, antioxidants are thought to play a pivotal role in preventing deleterious effects associated with exposure to such compounds. Ilex paraguariensis (IP) was used here to test the hypothesis that supplementation with the aqueous extract of this plant could exert protective effect against MMA or MA induced mortality, behavioral and/or biochemical changes in Drosophila melanogaster (DM). Initially, a curve time- and dose-response to MMA (1-10 mM), MA (1-10 mM) and IP (63-500 µM) was performed. Thereafter, flies were concomitantly exposed to MA (5 mM), MMA (5 mM) and/or IP (250 µg/mL) during 15 days for survival assay, and for 48 hs to MA (1 or 5 mM), MMA (1 or 5 mM) and/or IP (250 µg/mL) for subsequent investigations. Both MMA and MA exposure resulted in higher incidence of mortality, a worse performance in the negative geotaxis assay and increased locomotion in open-field test as compared with control group. Furthermore, a marked increase in non-protein thiol (NPSH) and in thiobarbituric acid reactive substances (TBARS) levels, decrease in superoxide dismutase (SOD), catalase and acetylcholinesterase (AChE) activities, and decrease in MTT and resazurin reduction were noted in MMA or MA treated groups. IP treatment offered significant protection against all alterations associated to MMA or MA exposure. This study confirm the hypothesis that supplementation with IP offers protection against changes associated to MMA or MA exposure in DM, due, at least in part, to its antioxidant effect.

Anti-inflammatory activity of herb products from Licania rigida Benth.

PURPOSE: The objective of the present study was to evaluate the systemic anti-inflammatory activity of the hydroalcoholic extract of the leaves of Licania rigida Benth (EHFLR) on models of systemic inflammation in mice. METHODS: The quantitative chemical profiles of phenolic acids and flavonoids were performed by High-Performance Liquid Chromatography (HPLC). Systemic anti-inflammatory activity was determined from carrageenan and dextran-induced paw edema models and the animals were orally treated (p.o.) with EHFLR at doses of 25, 50, 100 mg/kg, indomethacin (10 mg/kg) for carrageenan-induced paw edema and promethazine (6 mg/kg) for dextran-induced paw edema. The possible mechanisms involved in the anti-inflammatory action of the extract were evaluated by the paw edema models induced by histamine and arachidonic acid, and by the model of carrageenan-induced peritonitis, where vascular permeability and leukocyte migration to the peritoneal cavity were evaluated. RESULTS: The results of the HPLC identified the presence of phenolic acids and flavonoids, with chlorogenic acid (1.16%) and Caempferol (0.81%) as the main constituents. From the results, it was concluded that the extract has an LD50 ≥5000 mg/kg when administered orally in mice as this dose did not trigger deaths in any of the observed groups. EHFLR (25 mg/kg) showed a significant antiderematogenic effect on histamine and arachidonic acid-induced paw edema at the third hour of the tests, with a percentage of inhibition of 46.64% and 18.33%, respectively. The extract (25 mg/kg, p.o.) also significantly reduced vascular permeability and leukocyte migration in the peritoneal cavity. CONCLUSIONS: It is concluded that EHFLR exerts a systemic anti-inflammatory action, which seems to depend, at least in part, on the inhibition of arachidonic acid metabolism and the action of vasoactive amines. In addition, the extract reduced the leukocyte migration in the peritoneal cavity, indicating that its action may be linked to the inhibition of pro-inflammatory cytokines.

Comparative Analysis of the Antibacterial Activity and HPLC Phytochemical Screening of the Brazilian Red Propolis and the Resin of Dalbergia ecastaphyllum.

The aim of this study was to investigate the antibacterial activity of red propolis and resin and their association with standard antibiotics to evaluate possible differences of activity. We also submitted red propolis and the resin to a HPLC analysis to confirm the botanical origin. The extracts were tested against P. aeruginosa and S. aureus alone and in association with gentamicin and imipenem. The HPLC analysis identified seven compounds with six of them present in both substances. The lowest MIC values obtained in this study were observed against S. aureus. In general, MIC values showed to be lower for red propolis against all species tested in comparison to resin. Despite the synergistic behavior to be similar for both substances, we observed that inhibitory concentrations of drugs were lower when associated with red propolis in comparison to resin.

Chemical composition and anti-Candida potencial of the extracts of Tarenaya spinosa (Jacq.) Raf. (Cleomaceae).

Phytochemical prospecting was performed by HPLC-DAD. The Inhibitory Concentration of 50% of mortality the microorganisms (IC50) was determined and a cell viability curve was obtained. Minimum Fungicidal Concentration (MFC) was determined by subculture in Sabourad Dextrose Agar. The effect of the combination extract/fluconazole was verified by microdilution, with the extracts in subinhibitory concentrations (MFC/16). Caffeic acid was the major compound of both extracts, representing 6.08% in the aqueous extract and 7.62% in the ethanolic extract. The extracts showed a fungistatic effect (MFC ≥ 16,384 µg/mL). The IC50 results demonstrated that the combination of the extracts with fluconazole were more significant than the products tested alone, with values from 4.9 to 34.8 µg/mL for the ethanolic extract/fluconazole and 5 to 84.7 µg/mL for the aqueous extract/fluconazole. The potentiating effect of fluconazole action was observed against C. albicans and C. tropicalis. In C. krusei the aqueous extract had an antagonistic effect.

Ximenia americana L. enhances the antibiotic activity and inhibit the development of kinetoplastid parasites.

OBJECTIVE: The objective of this work was evaluate the cytotoxic, leishmanicidal and tripanocidal activity, as well as to evaluate its antimicrobial and modulatory activity in association with different antibiotics of the hydroethanolic extract of the Ximenia Americana stem bark (EHXA). METHOD: In vitro tests against Trypanosoma cruzi, Leishmania sp. and citotoxicity were performed. The evaluation of the antibacterial and bacterial resistance modulatory effect was given by the microdilution method. RESULTS: The chemical profile show different classes of compounds with significant presence of quercetrin and caffeic acid. The EHXA demonstrated activity only in the concentration of 1000 µg/mL against the L. infantum and L. brasiliensis promastigotes, causing mortality percentage of 40.66 and 27.62%, respectively. The extract presented a significant toxicity only in the concentration of 1000 µg/mL, causing a mortality of 55.42% of fibroblasts. The antibacterial activity of the EHXA demonstrated a MIC value ≥1024 µg/mL against all the tested bacteria. However, in the modulation assay with EHXA in association with different antibiotics the extract had a synergistic effect against S. aureus strains when associated with norfloxacin. CONCLUSION: The results of this investigation demonstrate for the first time the chemical composition of the hydroethanolic extract of the Ximenia Americana stem bark, your potential antiparasitic and modulatory effect. The low cytotoxic and biological potential against S. aureus open therapeutic perspectives against leishmaniosis and bacterial infections.

Antinociceptive activity of Copaifera officinalis Jacq. L oil and kaurenoic acid in mice.

Copaifera officinalis L. possesses traditional uses as an analgesic, anti-inflammatory, and antiseptic. However, until now the antinociceptive effect and the mechanism of action were not described for Copaifera officinalis L. oil and no compound present in this oil was identified to be responsible for its biological effects. The goal of this study was to identify the presence of kaurenoic acid in Copaifera officinalis oil and investigate its antinociceptive effect, mechanism of action, and possible adverse effects in mice. The quantification of kaurenoic acid in Copaifera officinalis oil was done by HPLC-DAD technique. Male and female albino Swiss mice (25-35 g) were used to test the antinociceptive effect of Copaifera officinalis (10 mg/kg, intragastric) or kaurenoic acid (1 mg/kg) in the tail-flick test, intraplantar injection of capsaicin, allyl isothiocyanate (AITC) or complete Freund's adjuvant (CFA). Copaifera officinalis oil and kaurenoic acid caused the antinociceptive effect in the tail-flick test in a dose-dependent manner, and their effect was reversed by naloxone (an opioid antagonist). Copaifera officinalis oil or kaurenoic acid reduced the nociception caused by capsaicin or AITC and produced an anti-allodynic effect in the CFA model (after acute or repeated administration for 7 days). Possible adverse effects were also observed, and non-detectable adverse effect was observed for the intragastric administration of Copaiba officinalis oil or kaurenoic acid and in the same way, the treatments were neither genotoxic nor mutagenic at the doses tested. Thus, Copaiba officinalis oil, and kaurenoic acid possess antinociceptive action without adverse effects.

GC-MS Chemical Characterization and In Vitro Evaluation of Antioxidant and Toxic Effects Using Drosophila melanogaster Model of the Essential Oil of Lantana montevidensis (Spreng) Briq.

Background and objectives: Natural products such as essential oils with antioxidant potential can reduce the level of oxidative stress and prevent the oxidation of biomolecules. In the present study, we investigated the antioxidant potential of Lantana montevidensis leaf essential oil (EOLM) in chemical and biological models using Drosophila melanogaster. Materials and methods: in addition, the chemical components of the oil were identified and quantified by gas chromatography coupled to mass spectrometry (GC-MS), and the percentage compositions were obtained from electronic integration measurements using flame ionization detection (FID). Results: our results demonstrated that EOLM is rich in terpenes with Germacrene-D (31.27%) and ß-caryophyllene (28.15%) as the major components. EOLM (0.12-0.48 g/mL) was ineffective in scavenging DPPH radical, and chelating Fe(II), but showed reducing activity at 0.24 g/mL and 0.48 g/mL. In in vivo studies, exposure of D. melanogaster to EOLM (0.12-0.48 g/mL) for 5 h resulted in 10% mortality; no change in oxidative stress parameters such as total thiol, non-protein thiol, and malondialdehyde contents, in comparison to control (p > 0.05). Conclusions: taken together, our results indicate EOLM may not be toxic at the concentrations tested, and thus may not be suitable for the development of new botanical insecticides, such as fumigants or spray-type control agents against Drosophila melanogaster.

Antinociceptive and anti-inflammatory activities of Philodendron bipinnatifidum Schott ex Endl (Araceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Popular medicine use stems of Philodendron bipinnatifidum (Araceae) in inflammation cases, such as in erysipelas, as well as orchitis and rheumatism treatment. The present study, conducted for the first time in literature, investigate the antinociceptive and anti-inflammatory activities of P. bipinnatifidum stems ethyl acetate extract (EPB). MATERIALS AND METHODS: GC/MS and HPLC analysis were performed for EPB extract. We used EPB at 250, 375 and 500 mg/kg (oral route, p.o.) in male Swiss mice. The antinociceptive activity of the plant extract assessed by acetic acid induced writhing and formalin tests. To investigate the possible participation of opioid system in EPB-mediated effects, we previously administered naloxone to the mice. Anti-inflammatory activity was evaluated using carrageenan-induced paw oedema. The open-field test aimed to investigate the possible EPB effects on the locomotor and exploratory activities. To assess the protective role of EPB on carrageenan-induced oxidative stress, the levels of NPSH, TBARS, as well as SOD and CAT activities were evaluated in blood and paw tissue. The acute toxicity of the EPB was investigated using OECD 423 guideline. RESULTS: The EPB chemical analysis by GC/MS and HPLC revealed the presence of flavonoids (luteolin and quercetin) and phytosterols (ß-sitosterol and stigmasterol). The oral treatment with the EPB inhibited mice abdominal writhings (P < 0.01) at 375 and 500 mg/kg, and reduced the formalin effect at the first-phase (500 mg/kg, P < 0.05) and also at the second-phase (500 mg/kg, P < 0.001) of the test. EPB (375 and 500 mg/kg) did not alter spontaneous locomotion in open field test, however the number of fecal bolus was significantly lower for the EPB group at 500 mg/kg when compared to the vehicle group (P < 0.05). The pretreatment with naloxone caused significant inhibition of antinociceptive activity induced by EPB in the formalin test, revealing the possible involvement of opioid receptors. EPB extract administered at 500 mg/kg (p.o.) prevented carrageenan-induced paw oedema (P < 0.05 and 0.01) until 6 h after carragenan injection. Evaluation of TBARS and NPSH levels, SOD and CAT activities in the blood and paw tissue of animals submitted to the carrageenan assay suggested that the anti-inflammatory effect of EPB may be linked to oxidative stress inhibition. The acute administration of the EPB (2000 mg/kg, p.o.) caused no mortality, demonstrating low toxicity. CONCLUSIONS: The extract of P. bipinnatifidum displays antinociceptive and anti-inflammatory activities, causing no toxicological effects. The pharmacological activity of this vegetal species may be related to the presence of flavonoids and phytosterols. Our results support the ethnomedical use of this vegetal species as analgesic and anti-inflammatory agent.

In vitro activities of Ceiba speciosa (A.St.-Hil) Ravenna aqueous stem bark extract.

Several species of the genus Ceiba (Malvaceae) are ethnopharmacologically used. Thus, this study aimed to investigate the in vitro beneficial properties of the aqueous stem bark extract of Ceiba speciosa. The extract presented a great amount of phenolic compounds (117.4 ± 6.2 mg GAE/g). The antioxidant activity was assessed by DPPH (IC50 = 42.87 µg/mL), ORAC (2351.17 µmol TE/g) and FRAP (235.94 µM FeSO4/g) methods. In addition, the extract reduced MCF-7 cell viability as assessed by MTT. However, it prevented mitochondrial membrane depolarization and reduced caspase-9 activity induced by hydrogen peroxide. In conclusion, these findings indicate the extract is an excellent source of natural antioxidants and is able to protect ROS-induced cell death. Therefore, C. speciosa extract may possess beneficial properties for application in pharmaceutical industry as an antioxidant. However, further studies to better elucidate its mechanisms and to isolate its active compounds are required.

Tetracarpidium conophorum Müll. Arg modulates sexual behaviour and biochemical parameters relevant to sexual function in male Wistar rats.

Walnut (Tetracarpidium conophorum Müll. Arg) has been reported to be an essential ingredient in folklore medicine for sexual enhancement with little scientific validation. Hence, this study investigated the effects of walnut supplemented diet on sexual behaviour and biochemical parameters relevant to erection in male Wistar rats. Forty animals used in this study were divided into five groups (n = 8); Group 1 - normal control rats fed with basal diet, Group II - rats fed diet supplemented with 10% processed walnut, Group III - rats fed diet supplemented with 10% raw walnut, Group IV - rats fed diet supplemented with 20% processed walnut and Group V - rats fed diet supplemented with 20% raw walnut. Behavioural studies (copulation tendency and anxiety) associated with sexual function, measurement of nitric oxide (NO) levels, adenosine deaminase (ADA), arginase and acetylcholinesterase (AChE) activities in the Corpus cavernosum as well as characterization of bioactive components of the nut were evaluated. Marked reductions in ADA and arginase activities and a concomitant increase (% inclusion dependent) in the level of NO as well as enhanced sexual behaviours were observed in rat fed supplemented walnut when compared to the control. Furthermore, analysis of the walnut using high performance liquid chromatography indicated the presence of some polyphenols. From our findings, it showed that walnut improves sexual behaviour and modulates activities of key enzymes relevant to erection in male rats which may justify its used in traditional medicine.