Chemical study of Adenocalymma axillarum crude leaf extract and isolated compounds

Abstract Adenocalymma axillarum (K.Schum.) L.G. Lohmann is a liana belonging to the family Bignoniaceae. In traditional medicine, the genus Adenocalymma is used to treat fever, skin ailments, and body, joint, and facial muscle pains, and it is also applied as cosmetic. Biological assays conducted with the A. axillarum crude leaf ethanol extract have indicated leishmanicidal activity and absence of cytotoxicity. This study aimed to analyze the A. axillarum leaf ethanol crude extract by high-performance liquid chromatography-high-resolution mass spectrometry- diode array detector (HPLC-HRMS-DAD) and to evaluate the leishmanicidal and cytotoxic activities of this crude extract, its fractions, and isolated compounds. HPLC-HRMS-DAD analysis of this extract revealed that it consisted mainly of flavonoids, with nine major compounds. Extract purification yielded 4-hydroxy-N-methylproline, 6-β-hydroxyipolamiide, quercetin-3-O-robinobioside, hyperin, isorhamnetin-3-O-robinobioside, and 3'-O-methylhyperin, which were identified by Nuclear Magnetic Resonance. The isolated compounds were inactive against Leishmania amazonensis promastigotes and human lung fibroblast cells.

Effect of salicylic acid and silver nitrate on rutin production by Hyptis marrubioides cultured in vitro / Efeito do ácido salicílico e nitrato de prata na produção de rutina por Hyptis marrubioides cultivada in vitro

ABSTRACT: Hyptis marrubioides (Lamiaceae) is a medicinal plant that is native from Brazilian Cerrado. In vitro propagation techniques make use of elicitors to alter metabolic pathways, affecting how molecules are produced both qualitatively and quantitatively. This research aimed to evaluate how abiotic elicitors salicylic acid (SA) and silver nitrate (SN) at concentrations of 30µM or 60µM influence Hyptis marrubioides seedling growth by two different in vitro culture methods. The rutin content was quantified by HPLC-DAD. Compared to an untreated culture, the H. marrubioides methanolic extracts cultured in MS medium for 10 days followed by culture in MS medium containing SN (30µM) for 20 days had 1.28 times higher rutin content. In a second experiment, seedlings were cultured in MS medium for 20 days, and then the desired elicitor was added to the culture and allowed to remain in contact with the medium for three and six days. SA (30µM) gave the best results: rutin production was 16.56-foldhigher than the control after six days. SN (30µM) increased the rutin content by 1.17-fold. At the two concentrations evaluated during the elicitation experiments, neither SA nor SN altered the growth parameters shoot length, leaf number, and fresh and dry weight of H. marrubioides seedlings grown in vitro as compared to the control. Based on these results, the abiotic elicitors SA and SN successfully provide Hyptis marrubioides with increased rutin content in vitro.

RESUMO: Hyptis marrubioides (Lamiaceae) é uma planta medicinal nativa do Cerrado brasileiro. Técnicas de propagação in vitro fazem uso de elicitores para alterar as vias metabólicas, afetando a produção de moléculas qualitativa e quantitativamente. Este trabalho teve como objetivo avaliar como os elicitores abióticos ácido salicílico (SA) e nitrato de prata (SN) nas concentrações de 30µM ou 60µM influenciam no crescimento de plântulas de Hyptis marrubioides por dois diferentes métodos de cultivo in vitro. O teor de rutina foi quantificado por CLAE-DAD. Em comparação com uma cultura não tratada, os extratos metanólicos de H. marrubioides cultivados em meio MS por 10 dias, seguidos de cultura em meio MS contendo SN (30µM) por 20 dias, apresentaram 1,28 vezes maior teor de rutina. Em um segundo experimento, as plântulas foram cultivadas em meio MS por 20 dias, e então o elicitor desejado foi adicionado à cultura e deixado em contato com o meio por três e seis dias. SA (30µM) forneceu os melhores resultados na produção de rutina, sendo 16,56 vezes maior do que o controle após seis dias. SN (30µM) aumentou o teor de rutina em 1,17 vezes. Nas duas concentrações avaliadas durante os experimentos de elicitação, nem SA nem SN alteraram os parâmetros de crescimento, como comprimento da parte aérea, número de folhas ou peso fresco e seco das plântulas de H. marrubioides cultivadas in vitro em relação ao controle. Com base nestes resultados, os elicitores abióticos SA e SN forneceram com sucesso Hyptis marrubioides in vitro com maior conteúdo de rutina.

Impact of light quality on flavonoid production and growth of Hyptis marrubioides seedlings cultivated in vitro

ABSTRACT Hyptis marrubioides Epling, Lamiaceae, a species from Brazilian Cerrado, has been used against gastrointestinal infections, skin infections, pain, and cramps. Herein, H. marrubioides seedlings were cultured in vitro under different wavelengths (white, blue, green, red, and yellow) with 50 µmol m-2 s-1 irradiance and a 16-h photoperiod. After 20 and 30 days of cultivation, shoot length, leaf number, fresh mass, and dry mass were evaluated. The flavonoid rutin content was determined by the HPLC-DAD method. The shoots were longer in plants cultivated under yellow (16.603 ± 0.790 cm, 1.8-fold), red (15.465 ± 0.461 cm, 1.7-fold), and green (14.677 ± 0.737 cm, 1.6-fold) lights than in control plants exposed to white light (9.203 ± 0.388 cm). The number of leaves increased in plants exposed to red (23.425 ± 1.138, 1.1-fold) and green (22.725 ± 1.814, 1.1-fold) lights, compared to control plants (20.133 ± 0.827). Fresh (0.665 ± 0.048 g, 1.2-fold) and dry (0.066 ± 0.005 g, 1.3-fold) mass of seedlings were the highest in seedlings grown under red light, compared to seedlings grown under white light (0.553 ± 0.048 and 0.028 ± 0.004, respectively). However, rutin production was higher under white (0.308 mg g-1 of dry weight) and blue lights (0.298 mg g-1 of dry weight). Thus, red light induces plant growth and increases leaf number and dry weight in in vitro-cultivated H. marrubioides, whereas blue and white lights promote the greatest rutin accumulation.

In vitro trypanocidal activity of the Egyptian plant Schinopsis lorentizii against trypomastigote and amastigote forms of Trypanosoma cruzi.

Chagas’ disease is a chronic illness caused by the protozoan Trypanosoma cruzi. According to estimates, approximately 16-18 million people are infected in Latin American. Plant extracts exhibit a wide variety of secondary metabolites and can play an important role in the discovery of new compounds with biological potential. The in vitro trypanocidal activity of the extracts obtained from six plant species collected in Egypt (Parkia africana, Parkia roxburgi, Lagerstromeia speciosa, Schinopsis lorentzii, Lagerstromeia indica, and Sapindus saponaria) was assayed against trypomastigote and amastigote forms of T. cruzi. The cytotoxic activity of the most active extract was also evaluated by conducting MTT assays. S. lorentzii and S. saponaria were the most active extracts against the trypomastigote form;IC50 values were 9.9 and 27.34 g/mL, respectively. The S. lorentzii extract was also evaluated against the amastigote form (IC50 was 111.5 g/mL).The S. lorentzii extract did not exhibit significant cytotoxic activity. The selectivity index value indicated that this extract was highly selective for the parasite. The S. lorentzii and S. saponariaextracts exhibit trypanocidal activity, probably as a result of the presence ofdifferent constituentsand their concentrations in the extracts.

Antimicrobial activity of the essential oil of Tetradenia riparia (Hochst.) Codd. (Lamiaceae) against cariogenic bacteria

In Brazilian folk medicine, Tetradenia riparia (Hochst.) Codd. (Lamiaceae) is used to treat toothaches and dental abscesses and diseases induced by worms, bacteria, or fungi. This paper aims to investigate the chemical composition and the antibacterial effects of the essential oil obtained from Tetradenia riparia leaves (TR-EO) grown in Southeastern Brazil against a representative panel of oral pathogens. We evaluated the antibacterial activity of TR-EO in terms of the minimal inhibitory concentration (MIC). We identified aromadendrene oxide (14.0%), (E,E)-farnesol (13.6%), dronabinol (12.5%), and fenchone (6.2%) as the major constituents of TR-EO. TR-EO displayed MIC values between 31.2 and 500 μg/mL, with the lowest MIC value being obtained against Streptococcus mitis (31.2 μg/mL), S. mutans (62.5 μg/mL), S. sobrinus (31.2 μg/mL), and Lactobacillus casei (62.5 μg/mL). In time-kill experiments, TR-EO demonstrated bactericidal activity against S. mutans within the first 12 h, resulting in a curve profile similar to that of chlorhexidine. These results revealed that the essential oil of Tetradenia riparia displays promising activity against most of the selected cariogenic bacteria, including Streptococcus mutans.

Evaluation of the antibacterial potential of Petroselinum crispum and Rosmarinus officinalis against bacteria that cause urinary tract infections

In this study we evaluated the antibacterial activity of the crude hydroalcoholic extracts, fractions, and compounds of two plant species, namely Rosmarinus officinalis and Petroselinum crispum, against the bacteria that cause urinary tract infection. The microdilution method was used for determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The crude hydroalcoholic extract of R. officinalis displayed in vitro activity against Gram-positive bacteria, with satisfactory MBC for the clinical isolate S. saprophyticus. The fractions and the pure compound rosmarinic acid did not furnish promising results for Gram-negative bacteria, whereas fractions 2, 3, and 4 gave encouraging results for Gram-positive bacteria and acted as bactericide against S. epidermidis as well as E. faecalis (ATCC 29212) and its clinical isolate. R. officinalis led to promising results in the case of Gram-positive bacteria, resulting in a considerable interest in the development of reliable alternatives for the treatment of urinary infections.

Antibacterial evaluation of Styrax pohlii and isolated compounds

The antibacterial activity of the compounds egonol (1) and homoegonol (2), of the crude ethanolic extract of Styrax pohlii (Styracaceae) aerial parts (EE), and of its n-hexane (HF), EtOAc (EF), n-BuOH (BF), and hydromethanolic (HMF) fractions was evaluated against the following microorganisms: Streptococcus pneumoniae (ATCC 6305), S. pyogenes (ATCC 19615), Haemophilus influenzae (ATCC 10211), Pseudomonas aeruginosa (ATCC 27853), and Klebsiella pneumoniae (ATCC 10031). The broth microdilution method was used for determination of the minimum inhibitory concentration (MIC) during preliminary evaluation of antibacterial activity. The EE yielded MIC values of 400 µg/mL for S. pneumoniae and P. aeruginosa and 300 µg/mL for H. influenzae. The HF and EF fractions exhibited enhanced antibacterial activity, with MIC values of 200 µg/mL against S. pneumoniae, but only EF displayed activity against H. influenzae (MIC 200 µg/mL). The best MIC value with compounds 1 and 2 (400 µg/mL) was obtained for (1) against S. pneumoniae and P. aeruginosa. Therefore, 1 exhibited weak antibacterial activity against these standard strains.

As atividades antimicrobianas das substâncias egonol (1) e homoegonol (2), do extrato etanólico das partes aéreas de Styrax pohlii (Styracaceae) (EE), bem como das frações n-hexano (HF), AcOEt (EF), n-BuOH (BF) e hidrometanólica (HMF) foram avaliadas frente aos seguintes microorganismos: Streptococcus pneumoniae (ATCC 6305), S. pyogenes (ATCC 19615), Haemophilus influenzae (ATCC 10211), Pseudomonas aeruginosa (ATCC 27853) e Klebsiella pneumoniae (ATCC 10031). O método de microdiluição em caldo foi utilizado para a determinação da concentração inibitória mínima (CIM) na avaliação preliminar da atividade antimicrobiana. EE mostrou valores de CIM de 400 µg/mL para S. pneumoniae e P. aeruginosa, e 300 µg/mL para H. influenzae. As frações HF e EF apresentaram melhora na atividade antimicrobiana, com valores de CIM de 200 µg/mL frente S. pneumoniae, mas apenas EF apresentou ação contra H. influenzae (200 µg/mL). Em relação às substâncias 1 e 2, o melhor valor de CIM (400 µg/mL) foi obtido por 1 frente a S. pneumoniae e P. aeruginosa, que exibiu fraca atividade antimicrobiana contra estas cepas padrões.

Determination of the antibacterial activity of crude extracts and compounds isolated from Hortia oreadica (Rutaceae) against oral pathogens / Determinação da atividade antibacteriana de extratos brutos e substâncias isoladas de Hortia oreadica (Rutaceae) frente à patógenos bucais

Extracts from Hortia oreadica afforded four dihydrocinnamic acid derivatives, isolated from the n-hexane extract, as well as limonoid guyanin and the furoquinoline alkaloid dictamnine, both isolated from the dichloromethane extract. The extracts and the isolated compounds were tested against some oral pathogens, so as to investigate their antibacterial activity. The results showed that the n-hexane extract and the compound dictamnine are the most active against the selected microorganisms.

Extratos brutos de Hortia oreadica, forneceram quatro derivados do ácido diidrocinâmico, que foram isolados do extrato n-hexânico, bem como as substâncias guianina e dictamina, isoladas do extrato em diclorometano. Os extratos brutos e as substâncias isoladas foram avaliados frente a alguns patógenos bucais com o objetivo de investigar a atividade antibacteriana. Os resultados demonstraram que o extrato bruto n-hexânico e a substância dictamina foram os mais ativos frente ao conjunto de microrganismos avaliados.

Atividade antibacteriana do extrato hidroalcoólico bruto de Stryphnodendron adstringens sobre microorganismos da cárie dental / Antibacterial activity of the crude hydroalcoholic extract of Stryphnodendron adstringens on dental caries microorganisms

Objetivo: O Stryphnodendron adstringens (barbatimão) é uma leguminosa do cerrado brasileiro, considerada como planta medicinal devido à sua ação cicatrizante, antiinflamatória, hemostática, anti-séptica e anti-diarréica. Este trabalho avaliou a atividade antibacteriana do extrato hidroalcoólico bruto do barbatimão contra microrganismos da cárie dental. Metodologia: Para determinação da Concentração Inibitória Mínima (CIM) do extrato hidroalcoólico bruto de barbatimão sobre as cepas padrão Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus e Lactobacillus casei foi utilizado o método da diluição em caldo. Resultados: O extrato hidroalcoólico bruto de barbatimão foi ativo frente aos microrganismos S. mitis e L. casei. A CIM do extrato do barbatimão variou de 350 a >400µg/mL. Conclusão: A ação antibacteriana observada sugere o uso do extrato de barbatimão como adjuvante no controle da cárie dental.

Purpose: Stryphnodendron adstringens ('barbatimão') is a medicinal plant of the Leguminoseae family from the Brazilian 'cerrado'. This plant is used in folk medicine because of its healing, anti-inflammatory, antiseptic, and hemostatic effects. The aim of this work was to evaluate the antibacterial activity of the 'barbatimão' crude hydroalcoholic extract against cariogenic microorganisms. Methods: The broth dilution method was used to determine the Minimum Inhibitory Concentration (MIC) of the 'barbatimão' hydroalcoholic extract against Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus casei. Results: The 'barbatimão' hydroalcoholic extract showed antibacterial activity against S. mitis e L. casei, with MIC ranging from 350 to >400µg/mL. Conclusion: These results suggest that the 'barbatimão' hydroalcoholic extract might be used as an adjuvant measure to control dental caries.

Antifeedant and allelopathic activities of the hydroalcoholic extract obtained from Neem (Azadirachta indica) leaves

Azadirachta indica, the so-called Neem or Nim, is well-known for its biological activities in many countries. The present study aimed at investigating the antifeedant properties of the hydroalcoholic extract obtained from the leaves of Neem on Zabrotes fasciatus (Coleoptera: Bruchidae), an insect pest that commonly feeds on common bean (Phaseolus vulgaris) during seed storage. We observed that the hydroalcoholic extract obtained from Neem leaves displayed significant antifeedant activity when it was added to the insect diet via Phaseolus vulgaris seeds. Additionally, the inhibitory effects of this extract on Phaseolus vulgaris germination and growth were also evaluated, indicating that both seed germination and radicle growth were affected in a concentration-dependent manner.

A espécie vegetal Azadirachta indica, popularmente chamada de Nim, é conhecida em muitos países devido a suas inúmeras propriedades biológicas. O presente estudo teve como objetivo investigar a propriedade antialimentar do extrato hidroalcoólico das folhas de Nim sobre Zabrotes fasciatus (Coleoptera: Bruchidae), um inseto conhecido como carucho-do-feijão e que atua como praga durante o processo de estocagem do feijão comum (Phaseolus vulgaris). Este inseto provoca danos tais como perda de peso, redução do valor nutritivo e do grau de higiene do feijão. Quando adicionado a sua dieta, usando sementes de Phaseolus vulgaris, o extrato hidroalcoólico das folhas de Nim demonstrou efeito antialimentar, ocasionando assim, uma significativa redução nos danos. Os efeitos inibitórios deste extrato sobre a germinação e o crescimento de Phaseolus vulgaris também foram avaliados. O extrato de Nim afetou a germinação das sementes e o crescimento das radículas de Phaseolus vulgaris de uma maneira dose-dependente.

Antimicrobial activity of Syzygium cumini (Myrtaceae) leaves extract

The antimicrobial activity of Syzygium cumini leaves extract, known as "jambolão", was evaluated. The crude hydroalcoholic extract was active against Candida krusei (inhibition zone of 14.7 ± 0.3 mm and MIC = 70 µg/mL), and against multi-resistant strains of Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus.

A atividade antimicrobiana do extrato das folhas de Syzygium cumini (Myrtaceae), conhecido como "jambolão", foi avaliada. O extrato hidroalcoólico mostrou atividade contra Candida krusei (halo de inibição de 14.7 ± 0.3 mm e CIM = 70 µg/mL) e cepas multiresistentes de Pseudomonas aeruginosa, Klebsiella pneumoniae e Staphylococcus aureus.

Evaluation of the in vitro antimicrobial activity of crude extracts of three Miconia species

A atividade antimicrobiana de nove extratos de três espécies de Miconia (M. albicans, M. rubiginosa e M. stenostachya) foi avaliada frente a onze microrganismos: Staphylococcus aureus, Staphylococcus saprophyticus, Proteus mirabilis, Escherichia coli, Enterococcus faecalis, Shigella flexneri, Klebsiella pneumoniae, Salmonella sp, Pseudomonas aeruginosa, Streptococcus agalactiae e Candida albicans. Três destes extratos apresentaram atividade antimicrobiana pelo método de difusão de poco. Os extratos etanólicos de Miconia albicans e Miconia rubiginosa foram os mais ativos.