The phytochemical study of Macrolobium acaciifolium extracts allowed the isolation and identifica... more The phytochemical study of Macrolobium acaciifolium extracts allowed the isolation and identification of 18 substances, 15 of which are reported for the first time in this species. From the methanolic extract of the leaves, the flavonoids luteolin-3'-O-α-L-rhamnoside (1), neoastilbin (2), astilbin (3), neoisoastilbin (4), isoastilbin (5), quercetin-3-O-α-L-rhamnoside (6), quercetin-3-O-rutinoside (7), kaempferol-3-O-rutinoside (8), quercetin-3-O-glicoside (9) and kaempferol-7-O-rutinoside (10) were isolated. From the methanolic extract of branches, the flavonoids chrysoeriol (13), apigenin (14), 7,4'-dihydroxyflavone (16), eriodictyol (17) and luteolin (18), in addition to the phenolic acids 3,4-dihydroxybenzoic acid (11), 3-methoxy,4-hydroxybenzoate (12) and methyl 3,4-dihydroxybenzoate (15) were isolated. The structural identification of the compounds was established by proton nuclear magnetic resonance (1 H NMR), carbon-13 nuclear magnetic resonance (13 C NMR), heteronuclear single quantum coherence (HSQC), heteronuclear multiple bond correlation (HMBC) and correlated spectroscopy (COSY) analyses. In this paper, we also discuss about the anisotropic effects on H-1'', H-2'', H-5'' and H-6'' rhamnose of isomers dihydroflavonoids.
Vismia japurensis Reichardt is a plant of ecological and chemical importance from which a variety... more Vismia japurensis Reichardt is a plant of ecological and chemical importance from which a variety of bioactive substances have been isolated. The current study aimed to establish in vitro cultures of this species as a source of secondary metabolites. Appropriate decontamination treatments and germination tests were performed and, after in vitro culture establishment, the propagated plants were multiplied in a sterile environment to increase the biomass of available experimental material. Seeds showed low contamination and a high germination percentage on Woody Plant Medium (WPM) supplemented with gibberellic acid (both at concentrations of 5 and 10 mg/L). V. japurensis nodal segments rapidly regenerated when first grown in WPM and then transplanted to Murashige and Skoog medium (MS). After 60 days in MS medium, the propagated plants were removed, lyophilized, and extracted with hexane and methanol. The hexane extract was fractionated via open column chromatography, and the substance isolated was purified by high performance liquid chromatography. Structural determination of the isolated substance was carried out using one and two-dimensional nuclear magnetic resonance and mass spectrometry. The isolated substance was identified as 1,8,10-trihydroxy-3,10-dimethyl-9(10H)-anthracenone, which, based on the conducted literature search, is reported for the first time.
The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to... more The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites. To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract. AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography. The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50) µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked. AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.
The phytochemical study of Macrolobium acaciifolium extracts allowed the isolation and identifica... more The phytochemical study of Macrolobium acaciifolium extracts allowed the isolation and identification of 18 substances, 15 of which are reported for the first time in this species. From the methanolic extract of the leaves, the flavonoids luteolin-3'-O-α-L-rhamnoside (1), neoastilbin (2), astilbin (3), neoisoastilbin (4), isoastilbin (5), quercetin-3-O-α-L-rhamnoside (6), quercetin-3-O-rutinoside (7), kaempferol-3-O-rutinoside (8), quercetin-3-O-glicoside (9) and kaempferol-7-O-rutinoside (10) were isolated. From the methanolic extract of branches, the flavonoids chrysoeriol (13), apigenin (14), 7,4'-dihydroxyflavone (16), eriodictyol (17) and luteolin (18), in addition to the phenolic acids 3,4-dihydroxybenzoic acid (11), 3-methoxy,4-hydroxybenzoate (12) and methyl 3,4-dihydroxybenzoate (15) were isolated. The structural identification of the compounds was established by proton nuclear magnetic resonance (1 H NMR), carbon-13 nuclear magnetic resonance (13 C NMR), heteronuclear single quantum coherence (HSQC), heteronuclear multiple bond correlation (HMBC) and correlated spectroscopy (COSY) analyses. In this paper, we also discuss about the anisotropic effects on H-1'', H-2'', H-5'' and H-6'' rhamnose of isomers dihydroflavonoids.
Vismia japurensis Reichardt is a plant of ecological and chemical importance from which a variety... more Vismia japurensis Reichardt is a plant of ecological and chemical importance from which a variety of bioactive substances have been isolated. The current study aimed to establish in vitro cultures of this species as a source of secondary metabolites. Appropriate decontamination treatments and germination tests were performed and, after in vitro culture establishment, the propagated plants were multiplied in a sterile environment to increase the biomass of available experimental material. Seeds showed low contamination and a high germination percentage on Woody Plant Medium (WPM) supplemented with gibberellic acid (both at concentrations of 5 and 10 mg/L). V. japurensis nodal segments rapidly regenerated when first grown in WPM and then transplanted to Murashige and Skoog medium (MS). After 60 days in MS medium, the propagated plants were removed, lyophilized, and extracted with hexane and methanol. The hexane extract was fractionated via open column chromatography, and the substance isolated was purified by high performance liquid chromatography. Structural determination of the isolated substance was carried out using one and two-dimensional nuclear magnetic resonance and mass spectrometry. The isolated substance was identified as 1,8,10-trihydroxy-3,10-dimethyl-9(10H)-anthracenone, which, based on the conducted literature search, is reported for the first time.
The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to... more The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites. To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract. AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography. The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50) µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked. AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.
Os micro-organismos endofíticos apresentam um enorme potencial para a produção de substâncias bio... more Os micro-organismos endofíticos apresentam um enorme potencial para a produção de substâncias bioativas. A planta Duroia macrophylla produz alcaloides com atividade antitumoral e antimicobacteriana, sendo selecionada para a pesquisa do potencial biotecnológico dos seus micro-organismos endofíticos. As folhas de D. macrophylla foram lavadas, submetidas à desinfecção, fragmentadas e inoculadas em BDA e Ágar Sabouraud com oxitetraciclina e incubadas a 30 °C por 20 dias. Isolaram-se 47 fungos e destes, 21 fungos morfologicamente diferentes foram selecionados para a prospecção química e biológica. O cultivo submerso foi realizado em Caldo Sabouraud ou Batata Dextrose, a 26 °C e 120 rpm durante 14-30 dias. Os líquidos metabólicos foram filtrados e submetidos à partição líquido-líquido com diclorometano e acetato de etila. Os metabólitos do micélio foram extraídos em ultrassom com diclorometano, acetato de etila e metanol. Os extratos e fases foram analisados por cromatografia em camada delgada e por ressonância magnética nuclear de 1H. Além disso, eles foram testados quanto às atividades antimicrobiana, antioxidante e contra Artemia salina. Como resultado deste trabalho a taxa de colonização fúngica de D. macrophylla foi de 94%. Nos 21 extratos foram encontrados indícios de substâncias fenólicas, terpenos, açúcares e em quatro deles, alcaloides. As fases AcOEt dos fungos Dm SB 43 e Dm BDA 12c foram ativas contra A. salina, com CL50 de 109,5 e 605,5 μg/mL, respectivamente. Nenhum extrato apresentou atividade antioxidante e 18 mostraram atividade antimicrobiana pelo menos para uma bactéria. O fungo Dm SB 33 apresentou o maior espectro de atividade, sendo ativo contra quatro bactérias: Enterobacter cloacae, Staphylococcus aureus, Acinetobacter baumanii e Pseudomonas aeruginosa. Apenas o fungo Dm SB 33 demonstrou atividade contra S. aureus e não foram observadas atividades contra E. coli. Portanto, este estudo demonstrou a diversidade de classes químicas e o potencial de atividades biológicas dos fungos endofíticos de D. macrophylla.
Micologia: fungos e/ou seus metabólitos como objeto de estudo, 2020
Os micro-organismos endofíticos apresentam um enorme potencial para a produção de substâncias bio... more Os micro-organismos endofíticos apresentam um enorme potencial para a produção de substâncias bioativas. A planta Duroia macrophylla produz alcaloides com atividade antitumoral e antimicobacteriana, sendo selecionada para a pesquisa do potencial biotecnológico dos seus micro-organismos endofíticos. As folhas de D. macrophylla foram lavadas, submetidas à desinfecção, fragmentadas e inoculadas em BDA e Ágar Sabouraud com oxitetraciclina e incubadas a 30 °C por 20 dias. Isolaram-se 47 fungos e destes, 21 fungos morfologicamente diferentes foram selecionados para a prospecção química e biológica. O cultivo submerso foi realizado em Caldo Sabouraud ou Batata Dextrose, a 26 °C e 120 rpm durante 14-30 dias. Os líquidos metabólicos foram filtrados e submetidos à partição líquido-líquido com diclorometano e acetato de etila. Os metabólitos do micélio foram extraídos em ultrassom com diclorometano, acetato de etila e metanol. Os extratos e fases foram analisados por cromatografia em camada delgada e por ressonância magnética nuclear de 1H. Além disso, eles foram testados quanto às atividades antimicrobiana, antioxidante e contra Artemia salina. Como resultado deste trabalho a taxa de colonização fúngica de D. macrophylla foi de 94%. Nos 21 extratos foram encontrados indícios de substâncias fenólicas, terpenos, açúcares e em quatro deles, alcaloides. As fases AcOEt dos fungos Dm SB 43 e Dm BDA 12c foram ativas contra A. salina, com CL50 de 109,5 e 605,5 μg/mL, respectivamente. Nenhum extrato apresentou atividade antioxidante e 18 mostraram atividade antimicrobiana pelo menos para uma bactéria. O fungo Dm SB 33 apresentou o maior espectro de atividade, sendo ativo contra quatro bactérias: Enterobacter cloacae, Staphylococcus aureus, Acinetobacter baumanii e Pseudomonas aeruginosa. Apenas o fungo Dm SB 33 demonstrou atividade contra S. aureus e não foram observadas atividades contra E. coli. Portanto, este estudo demonstrou a diversidade de classes químicas e o potencial de atividades biológicas dos fungos endofíticos de D. macrophylla.
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sendo selecionada para a pesquisa do potencial biotecnológico dos seus micro-organismos endofíticos. As folhas de D. macrophylla
foram lavadas, submetidas à desinfecção, fragmentadas e inoculadas em BDA e Ágar Sabouraud com oxitetraciclina e incubadas a 30 °C por 20 dias. Isolaram-se 47 fungos e destes, 21 fungos morfologicamente diferentes foram selecionados para a prospecção química e biológica. O cultivo submerso foi realizado em Caldo Sabouraud ou Batata Dextrose, a 26 °C e 120 rpm durante 14-30 dias. Os líquidos metabólicos foram filtrados e submetidos à partição líquido-líquido com diclorometano e
acetato de etila. Os metabólitos do micélio foram extraídos em ultrassom com diclorometano, acetato de etila e metanol. Os extratos e fases foram analisados por cromatografia em camada delgada e por ressonância magnética nuclear de 1H. Além disso, eles foram testados quanto às atividades antimicrobiana, antioxidante e contra Artemia salina. Como resultado deste trabalho a taxa de colonização fúngica de D. macrophylla foi de 94%. Nos 21 extratos foram encontrados
indícios de substâncias fenólicas, terpenos, açúcares e em quatro deles, alcaloides. As fases AcOEt dos fungos Dm SB 43 e Dm BDA 12c foram ativas contra A. salina, com CL50 de 109,5 e 605,5 μg/mL, respectivamente. Nenhum extrato apresentou atividade antioxidante e
18 mostraram atividade antimicrobiana pelo menos para uma bactéria. O fungo Dm SB 33 apresentou o maior espectro de atividade, sendo
ativo contra quatro bactérias: Enterobacter cloacae, Staphylococcus aureus, Acinetobacter baumanii e Pseudomonas aeruginosa. Apenas
o fungo Dm SB 33 demonstrou atividade contra S. aureus e não foram observadas atividades contra E. coli. Portanto, este estudo demonstrou a diversidade de classes químicas e o potencial de atividades biológicas dos fungos endofíticos de D. macrophylla.