RESUMO

Neglected tropical diseases are significant causes of death and temporary or permanent disability for millions living in developing countries. Unfortunately, there is no effective treatment for these diseases. Thus, this work aimed to conduct a chemical analysis using HPLC/UV and GC/MS to identify the major constituents of the hydroalcoholic extracts of Capsicum frutescens and Capsicum baccatum fruits, evaluating these extracts and their constituents' schistosomicidal, leishmanicidal and trypanocidal activities. The results obtained for the extracts of C. frutescens are better when compared to those obtained for C. baccatum, which can be related to the different concentrations of capsaicin (1) present in the extracts. The lysis of trypomastigote forms results for capsaicin (1) led to a significant value of IC50 = 6.23 µM. Thus, the results point to capsaicin (1) as a possible active constituent in these extracts.

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We report the chemical composition of the crude leaf extracts obtained from Stizophyllum perforatum (Cham.) Miers (Bignoniaceae), a simple high-performance liquid chromatography-diode array detection (HPLC-DAD) method based on mangiferin as an internal standard to quantify verbascoside, and the verbascoside acute oral toxicity and antileishmanial activity. HPLC-high-resolution mass spectrometry-DAD (HPLC-HRMS-DAD) analyses of the crude ethanol S. perforatum leaf extracts (CE-1 and CE-2) revealed that verbascoside was the major constituent in both extracts. CE-1 was purified, and verbascoside and casticin, among other compounds, were isolated. The developed HPLC-DAD method was validated and met the required standards. Investigation of the CE-2 acute toxicity indicated a lethal dose (LD50) greater than 2,000 mg/kg of body weight. Both CE-1 and CE-2 exhibited antileishmanial activity. The isolated compounds, verbascoside and casticin, also displayed antileishmanial activity with effective concentrations (IC50) of 6.23 and 24.20 µM against promastigote forms and 3.71 and 18.97 µM against amastigote forms of Leishmania amazonensis, respectively, but they were not cytotoxic to J774A.1 macrophages. Scanning electron microscopy of the L. amazonensis promastigotes showed that the parasites became more rounded and that their plasma membrane was altered in the presence of verbascoside. Additionally, transmission electron microscopy demonstrated that vacuoles emerged, lipids accumulated, kinetoplast size increased, and interstitial extravasation occurred in L. amazonensis promastigotes exposed to verbascoside. These findings suggest that S. perforatum is a promising candidate for further in vivo investigations against L. amazonensis.

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Urban vertical agriculture with lighting system can be an alternative green infrastructure to increase local food production irrespective of environmental and soil conditions. In this system, light quality control can improve the plant physiological performance, well as induce metabolic pathways that contribute to producing phenolic compounds important to human health. Therefore, this study aimed to evaluate the influence of RBW (red, blue and white) and monochromatic (red and blue; R and B, respectively) light associated or not with UV-B on photosynthetic performance and phenolic compound production in microtomato fruits cultivated via vertical agriculture. The experimental design adopted was completely randomized, with six replicates illuminated with 300 µmol·m-2·s-1 light intensities (RBW, RBW + UV, B, B + UV, R, and R + UV), 12 h photoperiod, and 3.7 W·m-2 UV-B irradiation for 1 h daily for the physiological evaluations. Twenty-six days after the installation, gas exchange, chlorophyll a fluorescence and nocturnal breathing were evaluated. Fruits in different ripening stages (green, orange, and red) were collected from microtomato plants grown under with different light qualities, to evaluate the physiological performance. The identification and quantification of the phenolic compound rutin was also performed to investigate their metabolic response. This study identified that plants grown under B + UV had high photosynthetic rates (A=11.57 µmol·m-2·s-1) and the fruits at all maturation stages from plants grown under B and B + UV had high rutin content. Meanwhile, the activation of suppressive mechanisms was necessary in plants grown under R because of the high nocturnal respiration and unregulated quantum yield of the non-photochemical dissipation of the photosystem II. These results highlight the importance of selecting light wavelength for vegetable cultivation to produce fruits with a high content of specialized metabolites that influence color, flavor, and health promotion, which is of special interest to farmers using sustainable cropping systems.

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Applying elicitors in plants growing under natural conditions change the metabolic content of volatile compounds extracted from their tissues. Here, we have investigated how abiotic elicitors influence the volatile composition of the ethyl acetate extract of Anacardium othonianum seedlings propagated in vitro and ex-vitro cultures. Abiotic factors such as salicylic acid, silver nitrate, light quality, and cultivation mode affected the composition of volatile compounds in ethyl acetate extract of A. othonianum leaves seedlings. GC/MS analysis demonstrated that α-tocopherol was the main metabolite in cultures added with chemical elicitors, reaching 22.48 ± 4.06% in the case of 30 µM salicylic acid. Higher concentrations of salicylic acid decreased the amount of α-tocopherol. The addition of AgNO3 had a linear effect on α-tocopherol. Thus, the production of this metabolite indicates that salicylic acid and AgNO3 elicitors positively affect α-tocopherol production at specific concentrations helping to understand how plant culture can be better manipulated.

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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.

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Brosimum gaudichaudii is a plant species with medicinal relevance due to its furanocoumarin accumulation. The accumulation of these compounds in the root promotes predatory extractivism, which threatens the conservation of the species. In addition, little is known about the conditions for culturing of this species in vitro. The present study aimed to investigate how the application of different spectra of LEDs (white, blue, red, and combinations of blue and red at 1:1 and 3:1 ratios) can impact the morphophysiological and biochemical characteristics of B. gaudichaudii under different in vitro conditions. To evaluate the production of furanocoumarins in its leaves, which are easy-to-collect perennial organs, we cultured nodal segments in 50-mL tubes with MS medium under 100 µmol m-2 s-1 light and a photoperiod of 16 h for 50 days. We then submitted the seedlings biometric, anatomical, biochemical, and physiological evaluations. The different spectral qualities influenced several characteristics of the seedlings. Plants grown under red light showed greater stem elongation and larger and thinner leaves, strategies aimed at capturing a higher ratio of radiant energy. Exposure to the blue/red ratio of 1:1 induced increases in the concentration of the furanocoumarin psoralen, probably due to the diversion of carbon from primary metabolism, which resulted in lower growth. Cultivation under blue light or blue:red light at 3:1 triggered anatomical and physiological changes that led to higher production of secondary metabolites in the leaves, and at the 3:1 ratio, the seedlings also had a high growth rate. These results highlight the fundamental role of light in stimulating the production of secondary metabolites, which has important implications for the production of compounds of interest and indirect consequences for the conservation of B. gaudichaudii.

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The genus Absidia is widely used in the biotransformation of different classes of natural products. This study evaluates the ability of the Absidia coerulea 3A9 marine derived strain isolated from the ascidian Distaplia stilyfera to perform biotransformations by conducting assays with (-)-cubebin, as substrate. The experiment was optimized using the experimental design proposed by Plackett-Burman for seven factors and eight experiments, to establish the biotransformation conditions that would allow maximum production of biotransformed dibenzylbutyrolactone (-)-hinokinin. An analytical method based on Reverse-Phase-High Performance Liquid Chromatography (RP-HPLC) was developed to quantify the fungal biotransformation product. The factor that influenced the (-)-hinokinin peak area the most positively was the percentage of seawater (%seawater) given that its %relative standard deviation (%RSD) showed a 32.92% deviation from the real value.

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Candidiasis therapy, especially for candidiasis caused by Candida non-albicans species, is limited by the relatively reduced number of antifungal drugs and the emergence of antifungal tolerance. This study evaluates the anticandidal activity of 41 plant-derived products against Candida species, in both planktonic and biofilm cells. This study also evaluates the toxicity and the therapeutic action of the most active compounds by using the Caenorhabditis elegans-Candida model. The planktonic cells were cultured with various concentrations of the tested agents. The Cupressus sempervirens, Citrus limon, and Litsea cubeba essential oils as well as gallic acid were the most active anticandidal compounds. Candida cell re-growth after treatment with these agents for 48 h demonstrated that the L. cubeba essential oil and gallic acid displayed fungistatic activity, whereas the C. limon and C. sempervirens essential oils exhibited fungicidal activity. The C. sempervirens essential oil was not toxic and increased the survival of C. elegans worms infected with C. glabrata or C. orthopsilosis. All the plant-derived products assayed at 250 µg/mL affected C. krusei biofilms. The tested plant-derived products proved to be potential therapeutic agents against Candida, especially Candida non-albicans species, and should be considered when developing new anticandidal agents.

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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.(AU)

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.(AU)

RESUMO

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.