Evaluation of the in vitro schistosomicidal, leishmanicidal, and trypanocidal activities of the capsaicin metabolite, Capsicum frutescens, and Capsicum baccatum extracts and of their analysis of the main constituents by HPLC/UV and CG/MS.

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.

HPLC method for quantifying verbascoside in Stizophyllum perforatum and assessment of verbascoside acute toxicity and antileishmanial activity.

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.

Light means power: harnessing light spectrum and UV-B to enhance photosynthesis and rutin levels in microtomato plants.

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.

Metabolic response induced by abiotic elicitors in Anacardium othonianum seedlings.

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.

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.

Streptomyces griseocarneus R132 expresses antimicrobial genes and produces metabolites that modulate Galleria mellonella immune system.

Actinobacteria is a phylum composed of aerobic, Gram-positive, and filamentous bacteria with a broad spectrum of biological activity, including antioxidant, antitumor, and antibiotic. The crude extract of Streptomyces griseocarneus R132 was fractionated on a C18 silica column and the isolated compound was identified by 1H and 13C nuclear magnetic resonance as 3-(phenylprop-2-enoic acid), also known as trans-cinnamic acid. Antimicrobial activity against human pathogens was assayed in vitro (disk-diffusion qualitative test) and in vivo using Galleria mellonella larvae (RT-qPCR). The methanol fractions 132-F30%, 132-F50%, 132-F70%, and 132-F100% inhibited the Escherichia coli (ATCC 25922) and Staphylococcus aureus (MRSA) growth in vitro the most effectively. Compared with the untreated control (60-80% of larvae death), the fractions and isolated trans-cinnamic acid increased the survival rate and modulated the immune system of G. mellonella larvae infected with pathogenic microorganisms. The anti-infection effect of the S. griseocarneus R132 fermentation product led us to sequence its genome, which was assembled and annotated using the Rast and antiSMASH platforms. The assembled genome consisted of 227 scaffolds represented on a linear chromosome of 8.85 Mb and 71.3% of GC. We detected conserved domains typical of enzymes that produce molecules with biological activity, such as polyketides and non-ribosomal and ribosomal peptides, indicating a great potential for obtaining new antibiotics and molecules with biotechnological application. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02942-1.

Combinations of Blue and Red LEDs Increase the Morphophysiological Performance and Furanocoumarin Production of Brosimum gaudichaudii Trécul in vitro.

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.

Optimization of (-)-cubebin biotransformation to (-)-hinokinin by the marine fungus Absidia coerulea 3A9.

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.

In Vitro and In Vivo Anti-Candida spp. Activity of Plant-Derived Products.

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.

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.

Genotoxic and Chemopreventive Effects of Vochysia divergens Leaves (Pantanal, Brazil).

The medicinal plant Vochysia divergens is a colonizing tree species of the Pantanal, a unique and little explored wetland region in Brazil. This species is used in folk medicine as syrups and teas to treat respiratory infections, digestive disorders, asthma, scarring, and skin diseases. The objectives of this study were to evaluate the antioxidant, cytotoxic, and genotoxic potential of the ethanolic extract of Vochysia divergens leaves (VdE), as well as the influence of VdE and its major component (the flavone 3',5-dimethoxy luteolin-7-O-ß-glucopyranoside; 3'5 DL) on MMS-induced genotoxicity. The extract significantly reduced the viability of V79 cells in the colorimetric XTT assay at concentrations ≥ 39 µg/mL. A significant increase in micronucleus frequencies was observed in V79 cell cultures treated with VdE concentrations of 160 and 320 µg/mL. However, animals treated with the tested doses of VdE (500, 1000, and 2000 mg/kg b.w.) exhibited frequencies that did not differ significantly from those of the negative control group, indicating the absence of genotoxicity. The results also showed that VdE was effective in reducing MMS-induced genotoxicity at concentrations of 20, 40, and 80 µg/mL in the in vitro test system and at a dose of 15 mg/kg b.w. in the in vivo test system. Its major component 3'5 DL exerted no protective effect, suggesting that it is not responsible for the effect of the extract. The results of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay showed that VdE was able to scavenge 92.6% of free radicals. In conclusion, the results suggest that the protective effect of VdE may be related, at least in part, to the antioxidant activity of its chemical constituents.

Diversity of cultivable fungal endophytes in Paullinia cupana (Mart.) Ducke and bioactivity of their secondary metabolites.

Paullinia cupana is associated with a diverse community of pathogenic and endophytic microorganisms. We isolated and identified endophytic fungal communities from the roots and seeds of P. cupana genotypes susceptible and tolerant to anthracnose that grow in two sites of the Brazilian Amazonia forest. We assessed the antibacterial, antitumor and genotoxic activity in vitro of compounds isolated from the strains Trichoderma asperellum (1BDA) and Diaporthe phaseolorum (8S). In concert, we identified eight fungal species not previously reported as endophytes; some fungal species capable of inhibiting pathogen growth; and the production of antibiotics and compounds with bacteriostatic activity against Pseudomonas aeruginosa in both susceptible and multiresistant host strains. The plant genotype, geographic location and specially the organ influenced the composition of P. cupana endophytic fungal community. Together, our findings identify important functional roles of endophytic species found within the microbiome of P. cupana. This hypothesis requires experimental validation to propose management of this microbiome with the objective of promoting plant growth and protection.

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.

18-Des-hydroxy Cytochalasin: an antiparasitic compound of Diaporthe phaseolorum-92C, an endophytic fungus isolated from Combretum lanceolatum Pohl ex Eichler.

Chemical investigation of the ethyl acetate extract from the endophytic fungus Diaporthe phaseolorum-92C (92C) isolated from the roots of Combretum lanceolatum led to the isolation of 18-des-hydroxy Cytochalasin H (compound 1). The trypanocidal and schistosomicidal activity and cytotoxicity of the extract from 92C were evaluated. The schistosomicidal, leishmanicidal, antimicrobial, and antioxidant actions, as well as the antitumor activity against the breast cancer cells MDA-MB-231 and MCF-7, and the cytotoxicity towards normal human lung fibroblasts GM07492A of compound 1 was tested. The extract from 92C (20 µg/mL) exerted potent trypanocidal activity, reducing 82% of the number of amastigotes and trypomastigotes of Trypanosoma cruzi. Compound 1 at 50 µg/mL killed 50% of Schistosoma mansoni adult worms. Compound 1 reduced the viability of Leishmania amazonenses promastigotes (IC50 = 9.2 µg/mL) and of the cancer cells MDA-MB-231 and MCF-7 (IC50 = 17.5 and 8.88 µg/mL, respectively), presented moderate antioxidant activity, and gave IC50 of 2049.7 ± 39.9 µg/mL for the cytotoxicity towards normal cells GM07492A. This knowledge is highly relevant to the search for new promising compounds for therapeutic purposes.

Bioactive compounds of Aspergillus terreus-F7, an endophytic fungus from Hyptis suaveolens (L.) Poit.

The compounds terrein (1), butyrolactone I (2), and butyrolactone V (3) were isolated from the ethyl acetate extract (EtOAc) of the endophytic fungus Aspergillus terreus-F7 obtained from Hyptis suaveolens (L.) Poit. The extract and the compounds presented schistosomicidal activity against Schistosoma mansoni; at 100 µg/mL for EtOAc extract, 1297.3 µM for compound 1, 235.6 µM for compound 2, and 454.1 µM for compound 3, they killed 100% of the parasites after 72 h of treatment. Compounds 1, 2, and 3 exerted moderate leishmanicidal activity against Leishmania amazonensis (IC50 ranged from 23.7 to 78.6 µM). At 235.6 and 227.0 µM, compounds 2 and 3, respectively, scavenged 95.92 and 95.12% of the DPPH radical (2,2-diphenyl-1-picryl-hydrazyl), respectively. Regarding the cytotoxicity against the breast tumor cell lines MDA-MB-231 and MCF-7, compound 2 gave IC50 of 34.4 and 17.4 µM, respectively, while compound 3 afforded IC50 of 22.2 and 31.9 µM, respectively. At 117.6 µM, compound 2 inhibited the growth of and killed the pathogen Escherichia coli (ATCC 25922). Compounds 1, 2, and 3 displayed low toxicity against the normal line of human lung fibroblasts (GM07492A cells), with IC50 of 15.3 × 103, 3.4 × 103, and 5.8 × 103 µM, respectively. This is the first report on (i) the in vitro schistosomicidal and leishmanicidal activities of the EtOAc extract of A. terreus-F7 and compounds 1, 2, and 3; and (ii) the antitumor activity of compounds 2 and 3 against MDA-MB-231 and MCF-7 cells.

Testing of Schefflera vinosa extract in mammalian cells in vitro and in vivo for potential toxicity, genetic damage, and role of oxidation.

The aim of this study was to examine the cytotoxic and genotoxic potential of a hydroethanolic extract of Schefflera vinosa (SV), a plant with schistosomicidal activity, as well as its influence on DNA damage induced by different mutagens, methyl methane sulfonate (MMS) and hydrogen peroxide (H2O2), in V79 cells and Swiss mice. Schefflera vinosa extract produced cytotoxicity at concentrations of 312.5 µg/ml or higher using the XTT cell proliferation assay kit. Treatment of V79 cell cultures with the highest SV concentration tested (150 µg/ml) significantly increased the frequency of micronuclei (MN) compared to controls. All SV concentrations significantly reduced the frequency of MN induced by hydrogen peroxide in V79 cell cultures. Further, SV was able to scavenge free radicals in the DPPH assay. In the in vivo test system, treatment with the highest dose tested (1,000 mg/kg body weight) induced a significant rise in frequency of DNA damage using the comet assay. However, animals treated with different doses of SV demonstrated absence of genotoxicity in the bone marrow MN test. For assessment of modulatory effects, the lower concentration of SV (250 mg/kg body weight) administered to MMS-treated mice significantly reduced frequency of DNA damage compared to the positive control (MMS alone). In contrast, the highest concentration tested (1,000 mg/kg body weight) significantly increased the rate of MN induced by MMS. The lack of genotoxic damage at biologically relevant SV concentrations, as well as the SV-mediated antigenotoxic and antioxidant activities, indicate the potential therapeutic usefulness of this plant extract. These activities may be attributed, at least in part, to the flavonoid quercitrin, its major component.

Quinone and hydroquinone metabolites from the ascidians of the genus Aplidium.

Ascidians of the genus Aplidium are recognized as an important source of chemical diversity and bioactive natural products. Among the compounds produced by this genus are non-nitrogenous metabolites, mainly prenylated quinones and hydroquinones. This review discusses the isolation, structural elucidation, and biological activities of quinones, hydroquinones, rossinones, longithorones, longithorols, floresolides, scabellones, conicaquinones, aplidinones, thiaplidiaquinones, and conithiaquinones. A compilation of the 13C-NMR spectral data of these compounds is also presented.

Antigenotoxicity of Roupala montana extract in the mouse micronucleus and comet assays.

Roupala montana Aubl. (Proteaceae) is a typical savannah species and native to tropical South America that has a moderate mortality for adult forms of Schistossoma mansoni. Because this species has been little studied, the aim of this investigation was to evaluate the influence of R. montana extract on DNA damage induced by methyl methanesulfonate (MMS) in peripheral blood cells and liver of Swiss mice using the micronucleus and comet assay, respectively. R. montana dichloromethane extract was prepared from a stock solution (0.5 mg/mL) in 5% dimethyl sulfoxide in water. Animals received a single dose of different concentrations of R. montana (5, 10 and 20 mg/kg body weight) by gavage (0.5 mL/animal). For antigenotoxicity assessment, different concentrations of R. montana were administered simultaneously with MMS diluted in water (40 mg/kg, intraperitoneally; 0.3 mL/animal). Peripheral blood and hepatocyte samples were obtained 48 and 24 h after treatment, respectively. Results showed that R. montana administered alone indicated the absence of genotoxicity in the mouse micronucleus or comet assay. On the other hand, administration of different doses of R. montana concomitantly with MMS led to a significant reduction in frequency of micronucleated polychromatic erythrocytes and DNA damage, when compared to the group treated only with MMS. Further, for the micronucleus assay, the gradual increase of R. montana concentration led to a proportional increase in the reduction of genotoxicity induced by MMS, indicating a dose-response relationship.

In vitro antiparasitic activity and chemical composition of the essential oil obtained from the fruits of Piper cubeba.

Protozoans of the trypanosomatid family cause the neglected tropical diseases leishmaniasis and trypanosomiasis, for which few drugs are available. In this context our group has recently reported that the essential oil obtained by steam distillation of the fruits of Piper cubeba is active against Schistosoma mansoni. Therefore, we have investigated the in vitro effects of the essential oil against the trypomastigote and amastigote forms of Trypanosoma cruzi isolated from an LLCMK2 cell line culture and the promastigote forms of Leishmania amazonensis. The in vitro activity of the essential oil against trypomastigotes of T. cruzi increased upon rising concentrations, giving IC50 values of 45.5 and 87.9 µg ·â€ŠmL⁻¹ against trypomastigotes and amastigotes, respectively. The essential oil was not active against L. amazonensis, since it displayed lyses of only 24 % at 400 µg ·â€ŠmL⁻¹, and an IC50 of 326.5 µg ·â€ŠmL⁻¹. Therefore, the essential oil should be further investigated to determine the compounds responsible for the observed activities, as well as its mechanism of action.

Chemical characterization and acaricide potential of essential oil from aerial parts of Tagetes patula L. (Asteraceae) against engorged adult females of Rhipicephalus sanguineus (Latreille, 1806).

Rhipicephalus sanguineus, commonly known as the brown dog tick, is one of the most widely distributed species of tick. In dogs, it can cause anemia and provide the transmission of pathogenic microorganisms such as Babesia canis, Ehrlichia canis, Hepatozoon canis, Anaplasma platys, and Mycoplasma haemocanis. To man, it can transmit the intracellular parasites Rickettsia rickettsii and Rickettsia conorii, the causative agents of the Rocky Mountain spotted fever in the Americas and Mediterranean and spotted fever in Europe and North Africa. Its control is performed by applying synthetic formulations composed of pyrethroids; however, continued use of these products results in environmental damage and acquisition of resistance. Alternatively, studies with botanical insecticides have been increasingly recurrent. Therefore, this study aimed to test the efficacy of essential oil of Tagetes patula, a ruderal species widely described in the literature for its insecticidal properties, in engorged females of R. sanguineus by the adults immersion test (AIT) and impregnated paper disk test (IPDT). The essential oil used, through gas chromatography coupled to mass spectrometry, revealed the presence of 55 compounds, being the 4-vinyl guaiacol and gamma terpinene the majority ones. The AIT compared to the IPDT was more efficient in inhibiting oviposition of tick; however, the eggs laid by the females submitted to saturated atmosphere with essential oil, from IPDT, not hatched, interrupted their development cycle. Besides being a pioneer work, the results presented here contributes to new researches, aiming the incorporation of essential oil in an acaricide for use in the environment.