Chromatographic analysis of selected phytosterols from Cyathea and their characterization by in silico docking to potential therapeutic targets.

Separation and quantification of lupeol, stigmasterol and swertiamarin in ethanolic extracts of selected Cyathea species have been developed using HPTLC and an attempt is made to explore the biopotential of phytochemicals against various proteins by computational analysis. Compounds were separated using the specific mobile phase and the developed plates were sprayed with respective spraying reagents. The 3D structure of the receptor proteins viz., 1VSN, 5BNQ, 6HN8, 7DN4 and 3TJU, and the 3D SDF structures of ligands like lupeol, stigmasterol and swertiamarin were retrieved from the Protein Data Bank (PDB) and NCBI-Pub Chem Compound database respectively. The Argus 4.0.1 is computer generated drug design screening software is employed to analyze the binding affinity of test compounds against the selected proteins in the form of E-values versus potential drug targets. The docking result was saved and visualized using Discovery Studio Visualizer. The terpenoid band with Rf value 0.79 depicted the presence of lupeol in C. gigantea (0.04%) and C. crinita (0.02%). The steroid band with Rf value 0.41 confirmed the presence of stigmasterol with varied frequency viz., C. nilgirensis (0.33%), C. gigantea (0.29%) and C. crinita (0.52%). Lupeol, stigmasterol and swertiamarin showed the interaction against the studied proteins viz., 1VSN, 5BNQ, 6HN8, 7DN4, 3TJU with varied energy values and interacting residues. The results of the virtual screening and molecular docking analysis suggest that the phytochemical compounds of Cyathea species viz., lupeol and stigmasterol were identified as possible lead molecules to fight against cancer and cytotoxicity.

Antinociceptive and anti-inflammatory activities of Hymenaea martiana Hayne (Fabaceae) in mice

Abstract Hymenaea martiana is a species popularly known in Northeastern Brazil as jatobá and used in folk medicine to treat pain and inflammation. The aim of this work was to evaluate the antinociceptive and anti-inflammatory activity of H. martiana. In the present study, we carried out an investigation about the effects of the crude ethanolic extract (Hm-EtOH) and the ethyl acetate fraction (Hm-AcOEt) in models of nociception and inflammation in mice. Chemical (acetic acid-induced writhing and formalin) and thermal stimuli (hot plate) were used for the evaluation of antinociceptive activity, while for the anti-inflammatory profile paw edema induced by carrageenan was used, along with leukocyte migration to the peritoneal cavity. The presence of the flavonoid astilbin in the samples was characterized through HPLC-DAD-MS analysis. Hm-EtOH and Hm-AcOEt (100, 200 and 400 mg.kg-1, i.p.) significantly reduced the number of abdominal contortions and decreased the paw licking time in the formalin test. In the hot plate, the extract increased the latency time of animals. Hm-EtOH and Hm-AcOEt inhibited significantly the increase in the edema after the administration of carrageenan. Hm-EtOH and Hm-AcOEt inhibited leukocyte migration in the peritonitis test. HPLC-DAD-MS analysis of Hm-EtOH and Hm-AcOEt revealed the presence of the flavonoid astilbin in the samples. According to the results of this study, both Hm-EtOH and Hm-AcOEt have antinociceptive and anti-inflammatory activities, which could be related with the presence of flavonoid in the extracts. The results reinforce the popular use of this plant.

Resumo Hymenaea martiana é uma espécie popularmente conhecida no Nordeste do Brasil como jatobá e usada na medicina popular para tratar a dor e a inflamação. O objetivo deste trabalho foi avaliar a atividade antinociceptiva e anti-inflamatória de H. martiana. No presente estudo, foram avaliados os efeitos do extrato etanólico bruto (Hm-EtOH) e da fração acetato de etila (Hm-AcOEt) em modelos de nocicepção e inflamação em camundongos. Foram utilizados estímulos químicos (contorções abdominais induzidas por ácido acético e teste da formalina) e estímulo térmico (teste da placa quente) para avaliação da atividade antinociceptiva, enquanto no perfil anti-inflamatório foi utilizado o teste do edema de pata induzido por carragenina e migração de leucócitos para a cavidade peritoneal. A presença do flavonoide astilbina nas amostras foi caracterizada através de análise por CLAE-DAD-EM. Hm-EtOH e o Hm-AcOEt (100, 200 e 400 mg.kg-1, i.p.) reduziram significativamente o número de contorções abdominais e diminuíram o tempo de lambida da pata no teste da formalina. No teste da placa quente, houve aumento do tempo de latência dos animais. Hm-EtOH e Hm-AcOEt inibiram significativamente o aumento do edema após a administração de carragenina, bem como inibiram a migração de leucócitos no teste de peritonite. A análise por CLAE-DAD-EM de Hm-EtOH e Hm-AcOEt revelou a presença do flavonoide astilbina nas amostras. De acordo com os resultados deste estudo, tanto Hm-EtOH quanto o Hm-AcOEt possuem atividades antinociceptiva e anti-inflamatória, o que pode estar relacionado à presença do flavonoide. Os resultados reforçam o uso popular desta planta.

Antinociceptive and anti-inflammatory activities of Hymenaea martiana Hayne (Fabaceae) in mice / Atividade antinociceptiva e anti-inflamatória de Hymenaea martiana Hayne (Fabaceae) em camundongos

Hymenaea martiana is a species popularly known in Northeastern Brazil as "jatobá" and used in folk medicine to treat pain and inflammation. The aim of this work was to evaluate the antinociceptive and anti-inflammatory activity of H. martiana. In the present study, we carried out an investigation about the effects of the crude ethanolic extract (Hm-EtOH) and the ethyl acetate fraction (Hm-AcOEt) in models of nociception and inflammation in mice. Chemical (acetic acid-induced writhing and formalin) and thermal stimuli (hot plate) were used for the evaluation of antinociceptive activity, while for the anti-inflammatory profile paw edema induced by carrageenan was used, along with leukocyte migration to the peritoneal cavity. The presence of the flavonoid astilbin in the samples was characterized through HPLC-DAD-MS analysis. Hm-EtOH and Hm-AcOEt (100, 200 and 400 mg.kg-1, i.p.) significantly reduced the number of abdominal contortions and decreased the paw licking time in the formalin test. In the hot plate, the extract increased the latency time of animals. Hm-EtOH and Hm-AcOEt inhibited significantly the increase in the edema after the administration of carrageenan. Hm-EtOH and Hm-AcOEt inhibited leukocyte migration in the peritonitis test. HPLC-DAD-MS analysis of Hm-EtOH and Hm-AcOEt revealed the presence of the flavonoid astilbin in the samples. According to the results of this study, both Hm-EtOH and Hm-AcOEt have antinociceptive and anti-inflammatory activities, which could be related with the presence of flavonoid in the extracts. The results reinforce the popular use of this plant.

Hymenaea martiana é uma espécie popularmente conhecida no Nordeste do Brasil como "jatobá" e usada na medicina popular para tratar a dor e a inflamação. O objetivo deste trabalho foi avaliar a atividade antinociceptiva e antiinflamatória de H. martiana. No presente estudo, foram avaliados os efeitos do extrato etanólico bruto (Hm-EtOH) e da fração acetato de etila (Hm-AcOEt) em modelos de nocicepção e inflamação em camundongos. Foram utilizados estímulos químicos (contorções abdominais induzidas por ácido acético e teste da formalina) e estímulo térmico (teste da placa quente) para avaliação da atividade antinociceptiva, enquanto no perfil anti-inflamatório foi utilizado o teste do edema de pata induzido por carragenina e migração de leucócitos para a cavidade peritoneal. A presença do flavonoide astilbina nas amostras foi caracterizada através de análise por CLAE-DAD-EM. Hm-EtOH e o Hm-AcOEt (100, 200 e 400 mg.kg-1, i.p.) reduziram significativamente o número de contorções abdominais e diminuíram o tempo de lambida da pata no teste da formalina. No teste da placa quente, houve aumento do tempo de latência dos animais. Hm-EtOH e Hm-AcOEt inibiram significativamente o aumento do edema após a administração de carragenina, bem como inibiram a migração de leucócitos no teste de peritonite. A análise por CLAE-DAD-EM de Hm-EtOH e Hm-AcOEt revelou a presença do flavonoide astilbina nas amostras. De acordo com os resultados deste estudo, tanto Hm-EtOH quanto o Hm-AcOEt possuem atividades antinociceptiva e anti-inflamatória, o que pode estar relacionado à presença do flavonoide. Os resultados reforçam o uso popular desta planta.

Antinociceptive and anti-inflammatory activities of Hymenaea martiana Hayne (Fabaceae) in mice / Atividade antinociceptiva e anti inflamatória de Hymenaea martiana Hayne (Fabaceae) em camundongos

Hymenaea martiana is a species popularly known in Northeastern Brazil as "jatobá" and used in folk medicine to treat pain and inflammation. The aim of this work was to evaluate the antinociceptive and anti-inflammatory activity of H. martiana. In the present study, we carried out an investigation about the effects of the crude ethanolic extract (Hm-EtOH) and the ethyl acetate fraction (Hm-AcOEt) in models of nociception and inflammation in mice. Chemical (acetic acid-induced writhing and formalin) and thermal stimuli (hot plate) were used for the evaluation of antinociceptive activity, while for the anti-inflammatory profile paw edema induced by carrageenan was used, along with leukocyte migration to the peritoneal cavity. The presence of the flavonoid astilbin in the samples was characterized through HPLC-DAD-MS analysis. Hm-EtOH and Hm-AcOEt (100, 200 and 400 mg.kg-¹, i.p.) significantly reduced the number of abdominal contortions and decreased the paw licking time in the formalin test. In the hot plate, the extract increased the latency time of animals. Hm-EtOH and Hm-AcOEt inhibited significantly the increase in the edema after the administration of carrageenan. Hm-EtOH and Hm-AcOEt inhibited leukocyte migration in the peritonitis test. HPLC-DAD-MS analysis of Hm-EtOH and Hm-AcOEt revealed the presence of the flavonoid astilbin in the samples. According to the results of this study, both Hm-EtOH and Hm-AcOEt have antinociceptive and anti-inflammatory activities, which could be related with the presence of flavonoid in the extracts. The results reinforce the popular use of this plant.

Hymenaea martiana é uma espécie popularmente conhecida no Nordeste do Brasil como “jatobᔠe usada na medicina popular para tratar a dor e a inflamação. O objetivo deste trabalho foi avaliar a atividade antinociceptiva e anti inflamatória de H. martiana. No presente estudo, foram avaliados os efeitos do extrato etanólico bruto (Hm-EtOH) e da fração acetato de etila (Hm-AcOEt) em modelos de nocicepção e inflamação em camundongos. Foram utilizados estímulos químicos (contorções abdominais induzidas por ácido acético e teste da formalina) e estímulo térmico (teste da placa quente) para avaliação da atividade antinociceptiva, enquanto no perfil anti-inflamatório foi utilizado o teste do edema de pata induzido por carragenina e migração de leucócitos para a cavidade peritoneal. A presença do flavonoide astilbina nas amostras foi caracterizada através de análise por CLAE-DAD-EM. Hm-EtOH e o Hm-AcOEt (100, 200 e 400 mg.kg-¹, i.p.) reduziram significativamente o número de contorções abdominais e diminuíram o tempo de lambida da pata no teste da formalina. No teste da placa quente, houve aumento do tempo de latência dos animais. Hm-EtOH e Hm-AcOEt inibiram significativamente o aumento do edema após a administração de carragenina, bem como inibiram a migração de leucócitos no teste de peritonite. A análise por CLAE-DAD-EM de Hm-EtOH e Hm-AcOEt revelou a presença do flavonoide astilbina nas amostras. De acordo com os resultados deste estudo, tanto Hm-EtOH quanto o Hm-AcOEt possuem atividades antinociceptiva e anti-inflamatória, o que pode estar relacionado à presença do flavonoide. Os resultados reforçam o uso popular desta planta.

Antinociceptive and anti-inflammatory activities of Hymenaea martiana Hayne (Fabaceae) in mice.

Hymenaea martiana is a species popularly known in Northeastern Brazil as "jatobá" and used in folk medicine to treat pain and inflammation. The aim of this work was to evaluate the antinociceptive and anti-inflammatory activity of H. martiana. In the present study, we carried out an investigation about the effects of the crude ethanolic extract (Hm-EtOH) and the ethyl acetate fraction (Hm-AcOEt) in models of nociception and inflammation in mice. Chemical (acetic acid-induced writhing and formalin) and thermal stimuli (hot plate) were used for the evaluation of antinociceptive activity, while for the anti-inflammatory profile paw edema induced by carrageenan was used, along with leukocyte migration to the peritoneal cavity. The presence of the flavonoid astilbin in the samples was characterized through HPLC-DAD-MS analysis. Hm-EtOH and Hm-AcOEt (100, 200 and 400 mg.kg-1, i.p.) significantly reduced the number of abdominal contortions and decreased the paw licking time in the formalin test. In the hot plate, the extract increased the latency time of animals. Hm-EtOH and Hm-AcOEt inhibited significantly the increase in the edema after the administration of carrageenan. Hm-EtOH and Hm-AcOEt inhibited leukocyte migration in the peritonitis test. HPLC-DAD-MS analysis of Hm-EtOH and Hm-AcOEt revealed the presence of the flavonoid astilbin in the samples. According to the results of this study, both Hm-EtOH and Hm-AcOEt have antinociceptive and anti-inflammatory activities, which could be related with the presence of flavonoid in the extracts. The results reinforce the popular use of this plant.

Evaluation of phytochemical composition, toxicity in Drosophila melanogaster and effects on antibiotics modulation of Plathymenia reticulata Benth extract.

Bacterial resistance is interfering with the action of antibiotics for clinical use in treating pathologies. The search for new substances capable of combating this resistance is necessary. An alternative to the search for these substances is in the extract of medicinal plants. Plathymenia reticulata, plant of the Fabaceae family, is a common tree species from the Brazilian cerrado, and is commonly used in areas of environmental degradation. This species is rich in phenolic compounds, such as flavonoids and tannins, compounds that are associated with various biological effects. A hydroethanolic extract from the bark of Plathymenia reticulata (HEPrB) was produced and then tests were carried out to verify the direct antibacterial activity, the modulatory effect of antibiotics for clinical use and their toxicity in Drosophila melanogaster flies. Through the analysis with UPLC, a wide variety of flavonoids contained in the HEPrB was observed. Direct antibacterial activity was observed for the standard strain of Staphylococcus aureus, however, the extract showed antagonistic activity or no significance in relation to the antibiotics tested in this study. As for toxicity, the HEPrB did not show significant damage in the proposed model. The results emphasize care when associating the consumption of teas with treatments with antibiotics for clinical use.

Gastroprotective and cicatrizing activity of the Ziziphus joazeiro Mart. leaf hydroalcoholic extract.

Ziziphus joazeiro Mart., popularly known as 'juazeiro', is a species used in popular medicine for the treatment of bronchitis, gastric ulcers, skin wounds, and in the manufacture of cosmetic and food products. The objective of this study is to evaluate the gastroprotective and cicatrizing activity of the Z. joazeiro Mart. leaf hydroalcoholic extract (EHFZJ). The acute pre-clinical toxicity was determined by the single administration of the EHFZJ (2000 mg/kg/p.o.) and by assessing clinical signs of toxicity, according to established criteria by Malone, or mortality. Gastroprotective activity was identified through classical models of acute gastric lesions induced by indomethacin, absolute and acidified ethanol (100, 200 and 400 mg/kg/per os) and the physical barrier mechanism (400 mg/kg/per os or intraperitoneally). The cicatrizing activity of the EHFZJ was investigated by measuring the speed of wound closure and the percentage of contraction. The acute pre-clinical toxicity of EHFZJ showed no signs of toxicity and mortality. The EHFZJ demonstrated a gastroprotective effect at the 400 mg/kg dose in the classical models of acute gastric injury induced by indomethacin, absolute and acidified ethanol. The EHFZJ administration (orally) demonstrated significant inhibition, suggesting a possible physical barrier mechanism exists. The EHFZJ showed no significant differences in terms of percentage of contraction or the speed of wound closure during the observation times (0, 3, 7, 11 and 14 days). The results obtained in this study provide evidence of a potential gastroprotective activity for the Ziziphus joazeiro Mart. Leaf hydroalcoholic extract.

The antioxidative effects of bioactive products from Sargassum polycystum C. Agardh and Sargassum duplicatum J. Agardh against inflammation and other pathological issues.

The present study was aimed to determine the phenol, total flavonoids and antioxidant potentials of Sargassum polycystum C. Agardh and Sargassum duplicatum J. Agardh from south east coast of Tamil Nadu and India using DPPH, phophomolybdenum and hydrogen peroxide scavenging activity. The total phenols, total flavonoids and antioxidant activities of S. polycystum and S. duplicatum were determined. Highest phenols (33.49 and 149.52 mg GAE/g) were observed in chloroform extracts of S. polycystum and methanolic extracts of S. duplicatum. The acetone extracts of S. polycystumand S. duplicatum demonstrated maximum amount of flavonoids compared to other studied extracts. Maximum amount of phosphomolybdenum reduction was observed in acetone extracts of S. polycystum and methanolic extracts of S. duplicatum. The DPPH radical scavenging activity of different extracts of S. polycystum and S. duplicatum were as follows acetone > chloroform > Petroleum ether > methanol and acetone > chloroform > methanol >Petroleum ether respectively. The H2O2 scavenging activity of S. polycystum extracts were as follows Petroleum ether extracts of S. polycystum (67.9%) > acetone (67.3%) > chloroform (58.6%) > methanol (51.78%). Acetone extracts of S. duplicatum showed maximum inhibition (90.39%) followed by petroleum ether, chloroform and methanolic extracts (75.11, 72.37 and 54.59%) respectively. The present study results confirmed the antioxidant properties of the two selected brown seaweeds viz., S. polycytum and S. duplicatum. The total phenols, flavonoids and alkaloids may be responsible for the antioxidant activities.

Antibacterial properties and modulation analysis of antibiotic activity of NaCe(MoO4)2 microcrystals.

This study reports the antibacterial properties and modulation analysis of antibiotic activity by NaCe(MoO4)2 microcrystals as well as their structural and morphological characterization. Evaluation of the antibacterial and antibiotic-modulating activity was carried out using the broth microdilution method. The Minimum Inhibitory Concentrations (MICs) of the compounds were expressed as the geometric mean of the triplicate values obtained through the use of Resazurin. Compound concentrations in the plates ranged from 512 to 0.5 µg/mL. Regarding its direct antibacterial activity, NaCe(MoO4)2 had a MIC ≥ 1024 µg/mL against all studied strains. As for its modulatory effect, it presented synergism with the antibiotic Gentamicin against the S. aureus strain and with Norfloxacin against E. coli, causing a reduction of 75% and 60%, respectively, in the antibiotic quantity required to have the same effect on the strain in study.

Larvicidal activity of some medicinal plant extracts against filariasis fever mosquito, Culex quinquefasciatus (Say.) (Diptera: Culicidae).

OBJECTIVES: The present study was aimed to find the larvicidal activity of petroleum ether, chloroform, acetone and methanolic extracts of Dichanthium foveolatum (Del.) Roberty, Leptochloa uniflora Hochst, Pancratium triflorum Roxb and Molineria trichocarpa (Wight) N.P.Balakr against Culex quinquefasciatus. METHODS: The larvicidal potential of selected plant extracts were determined against 4th instar larvae of C. quinquefasciatus with various concentrations viz., 50, 100, 150, 200 and 250 mg/ml. The mortality counts were made after 24 h of incubation and LC50 values were calculated. RESULTS: Chloroform extracts of studied plants were showed highest larvicidal activity with remarkable irritant against the larva of C. quinquefasciatus. Highest larvicidal activity was observed in the chloroform extract of D. foveolatum against the larva of C. quinquefasciatus with LC50 = 277.03 mg/ml. The larvicidal activity of the studied plants as follows chloroform extract of D. foveolatum (LC50 = 277.03 mg/ml) >L. uniflora (LC50 = 300.56 mg/ml) >M. trichocarpa (LC50 = 306.60 mg/ml) >P. triflorum (LC50 318.42 mg/ml). The larvicidal potential of P. triflorum was as follows Chloroform > acetone > methanol > petroleum ether. The larvicidal activities of L. uniflora and M. trichocarpa were as follows Chloroform > petroleum ether > acetone > methanol respectively. The larvicidal potentials of D. foveolatum was as follows Chloroform > methanol > acetone > petroleum ether. CONCLUSION: The chloroform extract of D. foveolatum find use as broad-spectrum larvicidal agent in the near future. It is hoped that more work would be undertaken to evaluate the utility of these plant extracts for field applications considering the promising leads given by the present study.

Modulation of antibiotic effect by Fe2(MoO4)3 microstrutures.

In this study, we report the antibacterial activity and modulation of antibiotic activity by Fe2(MoO4)3 microstructures obtained by the hydrothermal route without use of surfactants or organic additives. This material was characterized by X-ray diffraction (XRD), Raman spectroscopy and scanning electron microscopy (SEM) images. The XRD pattern showed that the Fe2(MoO4)3 crystallize in a monoclinic structure without secondary phases. Raman spectroscopy confirms the formation of Fe2(MoO4)3. SEM images show that the Fe2(MoO4)3 obtained have ball-of-yarn shaped morphology. In the antibacterial assays, strains of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were assayed by microdilution method to evaluate the antibacterial and modulatory-antibiotic activity with antibiotics as gentamicin, norfloxacin and imipenem. Against all bacteria, the Minimum Inhibitory Concentration (MIC) was Fe2(MoO4)3 ≥ 1024 µg/mL. This high MIC result must be associated with the fact of the iron be an essential microelement to the bacterial growth. However, when the Fe2(MoO4)3 was assayed in association with the antibiotics was observed an antagonistic effect demonstrated by an enhance of the MIC. This fact is associated directly with the pro-oxidative properties of metallic oxides. These compounds enhance the production of free radicals, as H2O2 and superoxide ions that can affect the cell structures as cell membrane and cell wall. Other effect is associated with the possible coordination of the metal, performing bonds with the chemical structure of the antibiotics, reducing their activity. Our results indicated that nanocompounds as Fe2(MoO4)3 can not be used as antimicrobial products for clinical usage, neither directly and neither in association with antibiotics.

Vertebrates as a Bactericidal Agent.

In Brazil, although a large number of animals are used in traditional medicine (at least 354 species), information about their biological activities is scarce. In this context, the objective of this study was to evaluate the bactericidal potential of zootherapeutic by-products from animals used in Brazilian traditional medicine and discuss the ecological and cultural consequences of such practices. The species analyzed were: Tupinambis merianae (skin), Iguana iguana (skin and body fat), Crotalus durissus (skin and body fat), Boa constrictor (skin), Euphractus sexcinctus (body fat) and Coendou prehensilis (quills). Experiments were performed with standard clinical strains of Escherichia coli (EC-ATCC10536) and Staphylococcus aureus (SA-ATCC 25923). For the microbiological assay, the zootherapeutics were evaluated using serial microdilutions. The results indicate that none of the samples possess inhibitory activity against standard bacterial strains. The in vitro ineffectiveness of the analyzed products demonstrate a necessity for new pharmacological research that encompass a large number of species of medicinal animals as well as highlight the importance of zootherapy in the context of plans for animal conservation.

Pancratium triflorum Roxb. (Amaryllidaceae) and Molineria trichocarpa (Wight) N.P. Balakr (Hypoxidaceae): Cytotoxic and antioxidant activities.

The present investigation was carried out to evaluate the antioxidant and cytotoxic activities of Pancratium triflorum Roxb and Molineria trichocarpa were collected from South Vagaikulam, Tirunelveli district, Tamil Nadu, India. The antioxidant activities of P. triflorum extracts were as follows with the IC50 values methanol (228.13 µg/mL) > chloroform (311.33 µg/mL) > acetone (398.08 µg/mL) > petroleum ether (410.16 µg/mL). The antioxidant activities of P. triflorum and M. trichocarpa extracts were as follows with the IC50 values methanol (80.93 µg/mL) > acetone (98.02 µg/mL) > chloroform (186.84 µg/mL) > petroleum ether (209.64 µg/mL). Among the various extracts of P. triflorum, methanolic extracts showed the strongest phosphomolybdenum reduction (140.56 g AA/100 g). Among the tested extracts, acetone extracts of M. trichocarpa showed maximum inhibition with 71.36 ±â€¯5.86%. In P. triflorum, chloroform extracts showed maximum inhibition (69.51%). The petroleum ether extract of M. trichocarpa was found to be most effective at which 50% mortality (LC50) and 90% mortality (LC90) of brine shrimp nauplii were found to be 29.22 and 184.82 mg/mL. This study results revealed the antioxidant and cytotoxic properties of P. triflorum and M. trichocarpa. Further investigations are needed to isolate and validate the active principles of the extract responsible various pharmacological properties.

Modulation of antibiotic activity by the hydroalcoholic extract from leaves of Caryocar coriaceum WITTM.

The aim of this study was to identify the presence of tannins, phenols and flavonoids on the hydroalcoholic extract of Caryocar coriaceum leaves (HECCL) and to determine the antioxidant and antibacterial activity of this extract. The extract was tested alone (1024-1 µg/mL) or associated (MIC/8) with several antibiotics in order to identify any antibacterial activity against multiresistant bacterial strains (Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa). The existence of tannins, total phenols (901.31 mg/g) and flavonoids (89.68 mg/g) was confirmed in the HECCL. The presence of rutin and quercetin were confirmed by Thin-layer chromatography (TLC). Using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, the antioxidant activity of the extract (9 µg/mL) was determined. Moreover, the Minimum Inhibitory Concentration (MIC) value found for HECCL was 1024 µg/mL and the association between HECCL (MIC/8) with benzylpenicillin significantly changed its minimum inhibitory concentration from 2500 to 625 µg/mL against E. coli.

Cholesterol and ergosterol affect the activity of Staphylococcus aureus antibiotic efflux pumps.

The aim of this study is to evaluate the effect of ergosterol on steroids and cholesterol efflux pumps in multidrug resistant strains of S. aureus. Were used RN4220 harboring plasmid pUL5054, which carries the gene encoding the MsrA macrolide efflux protein; and IS-58, which possesses the TetK tetracycline efflux protein; 1199B resists hydrophilic fluoroquinolones via a NorA-mediated mechanism and wild strain 1199B. The Minimal Inhibitory Concentration (MIC) was determined and the evaluation of possible inhibition of efflux pumps by reduction of MIC. Some of the detrimental effects on bacterial cells can be attributed to the detergent properties of cholesterol and ergosterol on account of their amphipathic structure. Besides the cholesterol did not affect directly the pump structure, a synergism was observed, maybe due the interaction with the cell membrane and interference in the lipid bilayer.

ß-Ag2MoO4 microcrystals: Characterization, antibacterial properties and modulation analysis of antibiotic activity.

This study reports the antibacterial properties and modulation analysis of antibiotic activity by ß-Ag2MoO4 microcrystals as well as their structural and vibrational characterization. The silver molybdate was obtained by the conventional hydrothermal method, and the structural, vibrational and morphological properties of the sample were determined using X-ray diffraction, Raman spectroscopy and scanning electron microscopy images. ß-Ag2MoO4 microcrystals obtained show spinel-type cubic structure (Fd-3m) with irregular shapes. The evaluation of antibacterial and modulatory-antibiotic activity was performed using the microdilution method to determine the Minimum Inhibitory Concentration (MIC) of the ß-Ag2MoO4 and antibiotics alone and associated with the silver molybdate. The ß-Ag2MoO4 modulates the antibiotic activity against all bacteria assayed in a synergistic (as the norfloxacin and gentamicin against S. aureus and gentamicin against E. coli) or an antagonistic form (as the norfloxacin against E.coli and P. aeruginosa). The reversion of antibiotic resistance by combinations with Ag2MoO4 could be a novel strategy to combat infections caused by multiple drug resistance (MDR) pathogens. Our results indicate that these silver molybdates present a clinically relevant antibacterial activity and enhanced the antibiotic activity of some antibiotics against MDR strain of S. aureus and E. coli, being an interesting alternative to combat antibiotic-resistant bacterial infectious agents.

Inhibition of the NorA efflux pump of Staphylococcus aureus by synthetic riparins.

AIM: The goal of this study was to increase knowledge about the antimicrobial activity of some synthetic Riparin-derived compounds, alone or in combination with fluoroquinolone antibiotics, against a strain of Staphylococcus aureus resistant to fluoroquinolone by way of overexpression of the NorA efflux pump. METHODS AND RESULTS: Microdilution tests showed that Riparins A and B did not show any significant antibacterial activity against Staph. aureus strains. On the other hand, the intrinsic antibacterial activity increased with increasing lipophilicity of the compounds, in the following order: Riparin-D (MIC 256 µg ml-1 ; Log P 2·95) < Riparin-C (MIC 102 µg ml-1 ; Log P 3·22) < Riparin-E (MIC 16 µg ml-1 ; Log P 3·57). The addition of all riparins to growth media at subinhibitory concentrations caused an increase in the antibacterial activity of antibiotics against the NorA-overexpressing test strain. Riparin-B, which has two methoxyl groups at the phenethyl moiety, showed the best modulatory effect. CONCLUSIONS: Riparin-E is a good anti-staphylococci agent, while Riparin-B functions as a NorA efflux pump inhibitor. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data suggest the possibility of using Riparin-B in combination with norfloxacin or ciprofloxacin for therapy of infections caused by multi-drug resistant Staph. aureus.

Análise fitoquímica e atividade antifúngica do óleo essencial de folhas de Lippia sidoides Cham. e do Timol contra cepas de Candida spp / Phytochemical analysis and antifungal activity of the essential oil of Lippia sidoides Cham. and of the Thymol against Candida strains

RESUMO A resistência de fungos do gênero Candida aos fármacos químicos tem lançado o desafio de se identificar novas substâncias que possuam atividade antibiótica ou venham a modular o efeito de produtos atualmente usados contra candidíase. O presente estudo avaliou a atividade antifúngica do óleo essencial de Lippia sidoides Cham. e do timol, sobre cepas de Candida. Inicialmente os produtos foram testados frente a 16 cepas fúngicas pela técnica de difusão em meio sólido, o que permitiu selecionar linhagens para continuidade da pesquisa. Com as linhagens de Candida krusei (CK LMBM 01, CK LMBM 02), Candida albicans (CA LM 62) e Candida tropicalis (CT LM 20), procedeu-se, por microdiluição em caldo, a determinação da Concentração Inibitória Mínima (CIM) e em meio sólido, a Concentração Fungicida Mínima (CFM) dos produtos foi identificada. O microcultivo das leveduras em meio empobrecido foi realizado para verificação de alterações morfológicas e, além disso, uma análise da composição química do óleo foi realizada por Cromatografia Gasosa acoplada à espectrometria de massas (CG-EM). Nesta análise, o constituinte majoritário foi o timol (84,95%), seguido de compostos como p-cimeno e Éter metil carvacrol, entre outros. A CIM do óleo essencial de Lippia sidoides Cham. frente às cepas variou entre 64 e 256 μg/mL, enquanto a CIM do timol foi estabelecida entre 32 e 64 μg/mL. A CFM do óleo essencial foi determinada entre 128 e 512 μg/mL e para o timol foram encontrados valores entre 64 e 128 μg/mL. Em relação à análise micromorfológica, verificada nas concentrações de CIM e CIM x 2, o óleo essencial inibiu o dimorfismo das cepas CK 01 e CT 20 na CIM e quando foi ensaiado o timol, este, na CIM, impediu a transição morfológica das cepas CK 01 e CA 62. Uma redução da morfogênese também foi obsevada na cepa CT 20, porém apenas em CIM x 2 e de forma mais discreta. Os resultados enaltecem o potencial antifúngico de L. sidoides e de seu composto majoritário timol tanto no combate à Candida quanto na neutralização de um dos fatores de virulência, a capacidade invasiva por formação de hifas e pseudohifas verificado na condição patogênica da candidíase. Estes dados são promissores e poderão incentivar futuras pesquisas sobre os aspectos fitoquímicos, toxicológicos e farmacológicos tanto do óleo essencial de Lippia sidoides como também de seus componentes químicos.

ABSTRACT The resistance of the Candida against drugs has been a challenge to the discovery of new substances with antimicrobial or modulatory effects that could be used against the cadidiasis. This work evaluated the antifungal activity of the essential oil of Lippia sidoides Cham. and of the Thymol against Candida strains. The products were tested towards 16 strains of Candida using the diffusion method, which allowed to select the strains in order to proceed with the research. The strains of Candida krusei (CK LMBM 01, CK LMBM 02), Candida albicans (CA LM 62) and Candida tropicalis (CT LM 20) were assayed by the microdilution method so that the Minimal Inhibitory Concentration (MIC) and the Minimal Fungicide Concentration (MFC) could be determined. The morphogenesis of the Candida was evaluated using poor environment in order to observe morphological changes. The composition of the essential oil was determined by GC-MS. The main compound observed was the thymol (84.95%). The MIC of the essential oil of L. sidoides and Thymol ranged between 64 to 256 μg/mL, and between 32 to 64 μg/mL respectively. The MFC of the essential oil and the thymol varied between 128 to 512 μg/mL and 64 to 128 μg/mL respectively. The morphogenesis of different Candida strains was inhibited in the MIC and MICx2 to the essential oil and thymol. The results indicated the antifungal potential of the L. sidoides and of the Thymol due to the inhibition of the invasive capacity, one of the most important virulence factors for the candidiasis` development. These results are promising to new researches about the phytochemical, toxicological and pharmacological aspects of the essential oil of L. sidoides and its phytochemical compounds.

Antimicrobial effect of Lippia sidoides and thymol on Enterococcus faecalis biofilm of the bacterium isolated from root canals.

The species Lippia sidoides Cham. (Verbenaceae) is utilized in popular medicine as a local antiseptic on the skin and mucosal tissues. Enterococcus faecalis is the bacterium isolated from root canals of teeth with persistent periapical lesions and has the ability to form biofilm, where it is responsible for the failure of endodontic treatments. Essential oil of L. sidoides (EOLS) and its major component, thymol, were evaluated for reducing the CFU in biofilms of E. faecalis in vitro. The essential oil was obtained by hydrodistillation and examined with respect to the chemical composition, by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis has led to the identification of thymol (84.9%) and p-cymene (5.33%). EOLS and thymol reduced CFU in biofilms of E. faecalis in vitro (time of maturation, 72 h), with an exposure time of 30 and 60 min at concentrations of 2.5 and 10%. There was no statistical difference in effect between EOLS and thymol, demontrating that this phenolic monoterpene was the possible compound responsible for the antimicrobial activity of EOLS. This study provides a basis for the possible utilization of EOLS as an adjuvant in the treatment of root canals that show colonization by E. faecalis.

Avaliação das potenciais atividades tripanocida e antileishmania do extrato de folhas de Piper arboreum (Piperaceae) e de suas frações / Evaluation of the potential trypanocidal and antileishmanial activities in Piper arboreum (Piperaceae) leaf extract and its fractions

A leishmaniose e a tripanossomíase americana fazem parte de um grupo de doenças tropicais endêmicas, especialmente entre as populações pobres. São consideradas doenças negligenciadas por não despertarem interesse da indústria farmacêutica. Atualmente, a quimioterapia é o único tratamento específico disponível para estas doenças, onde os medicamentos utilizados são nifurtimox e benzonidazol. Este trabalho teve como objetivo avaliar a atividade antiparasitária e citotóxica do extrato bruto etanólico e frações de folhas de Piper arboreum, frente a linhagens de Leishmania brasiliensis e de Trypanosoma cruzi. Após a preparação do extrato etanólico bruto e as suas respectivas frações, testes in vitro foram realizados para avaliar atividade antiparasitária frente a T. cruzi, utilizado o clone epimastigota CL-B5 e contra L.brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos NCTC929. Os resultados indicaram que as amostras apresentaram toxidade elevada, com exceção da fração de acetato de etila. Já os resultados da atividade antiparasitária indicaram uma maior atividade da fração hexânica observando a morte de 62% e 51% de células epimastigotas e de 100% e 92% de células promastigotas, respectivamente. Através dos resultados conclui-se que a P. arboreum pode ser considerada uma fonte alternativa de produtos naturais com atividade contra T. cruzi e L. brasiliensis...

Leishmaniasis and South American trypanosomiasis (Chagas disease) are part of a group of related human diseases endemic in tropical regions, especially among the poor. They are referred to as neglected diseases, because of a lack of interest on the part of the pharmaceutical industry. Today, chemotherapy is the only specific treatment against these diseases, the most frequently used drugs being nifurtimox and benznidazole. The aim of this study was to assess the antiparasitic and cytotoxic activity of the crude ethanolic leaf extract of Piper arboreum and its fractions against strains of Leishmania brasiliensis and Trypanosoma cruzi. After preparing the crude ethanolic extract and fractions, in vitro tests were performed to assay antiparasitic activity against T. cruzi, using the epimastigote clone CL-B5, while against L. brasiliensis, promastigote forms were used. The cytotoxicity assay was performed with a fibroblast cell line, NCTC 929. The results indicate that the samples possessed high toxicity, except for the ethyl acetate fraction. The results for antiparasitic activity showed highest activity in the hexane fraction, which was lethal to 62% and 51% of the epimastigotes and 100% and 92% of the promastigotes, at 500 and 100µg/mL, respectively. From these results, it is concluded that P. arboreum can be considered an alternative source of natural products with activity against T. cruzi and L.brasiliensis...