Assessment of the in Vitro Antimelanoma Potential of Lippia microphylla Cham (Verbenaceae) Essential Oil.

Essential oils stand out among natural products for their complex composition, frequently described in the literature with a range of biological effects. This study evaluated the cytotoxic activity against several human cancer cell lines of essential oils extracted from the leaves of Lippia microphylla (EO-LM) Cham. (Verbenaceae). The melanoma cell line SK-MEL-28 was the most sensitive to the EO-LM, presenting an IC50 of 33.38±1.16 µg/mL. Afterward, the effects of EO-LM on the cell cycle, induction of apoptosis, and production of reactive oxygen species (ROS) were evaluated. We stated a significant increase in the sub-G1 population, indicating apoptosis, later confirmed by an increase of SK-MEL-28 cells labeled with Annexin V-FITC and by the formation of apoptotic bodies and membrane blebs, observed by confocal microscopy. Additionally, EO-LM reduced the production of ROS, indicating antioxidant activity. Therefore, EO-LM exhibits anti-melanoma activity in vitro, suggesting its potential as an anticancer agent.

Phenolic-rich extract of Nopalea cochenillifera attenuates gastric lesions induced in experimental models through inhibiting oxidative stress, modulating inflammatory markers and a cytoprotective effect.

Nopalea cochenillifera (Cactaceae), popularly known as "palma" or "palma doce", is from Mexico, but it was widely introduced in Brazil through crops. It has been used as food and in traditional medicine and is a good source of phenolic compounds. In this study the phytochemical profile and gastroprotective activity of phenolic-rich extract of N. cochenillifera in acute gastric lesion models induced by ethanol and indomethacin were evaluated. High-performance liquid chromatography coupled with mass spectrometry (HPLC/ESI/MSn) allowed the characterization of 12 compounds such as sugars, phenolics and flavonoids. Among polyphenols, the main peak was assigned to isorhamnetin-3-O-(2'',3''-O-di-rhamnose)-glucoside. The TPC and TFC in the dry extract were 67.85 mg of gallic acid equivalent per g/extract and 46.16 mg quercetin equivalent per g/extract, respectively. In the in vitro MTT assay, the extract showed no cytotoxicity and suppressed ROS levels in LPS-treated RAW 264.7 cells. Preclinical models in rats showed that a dose of 100 mg kg-1 (p < 0.0001) in the ethanol model and doses of 100 mg kg-1 (p < 0.5) and 200 mg kg-1 (p < 0.01) in the indomethacin model reduced the gastric lesions. Also, the extract reduced the MPO, MDA, TNF-α and IL-1ß levels and increased the GSH and IL-10 levels. The pre-treatment with the extract led to the upregulation of SOD and the downregulation of COX-2 by immunohistochemical analysis. It also showed a cytoprotective effect in the histopathological analysis and stimulated the restoration of the mucus content as observed in the periodic acid-Schiff analysis without modifying the pH, volume or total acidity of the gastric juice. Taken together, N. cochenillifera extract can be applied as a novel gastroprotective ingredient for food or pharmaceutical products.

Nutraceutical formulations combining Limosilactobacillus fermentum, quercetin, and or resveratrol with beneficial impacts on the abundance of intestinal bacterial populations, metabolite production, and antioxidant capacity during colonic fermentation.

This study evaluated the impacts of different nutraceutical formulations combining Limosilactobacillus fermentum 296 (∼10 log CFU/mL), quercetin (QUE, 160 mg), and or resveratrol (RES, 150 mg) on the relative abundance of various intestinal bacterial populations, production of microbial metabolites, and antioxidant capacity during 48 h of in vitro colonic fermentation. The nutraceutical formulations increased the relative abundance of Lactobacillus spp./Enterococcus spp. and Bifidobacterium spp. and decreased the relative abundance of Bacteroides spp./Prevotella spp., Clostridium histolyticum, and E. rectale/C. coccoides during the colonic fermentation. Medium with the formulation containing L. fermentum, QUE, and RES had the highest prebiotic indexes, indicating synergistic or additive interaction between QUE and RES to modulate the intestinal microbiota. The nutraceutical formulations increased the production of bioactive metabolites and antioxidant capacity in the colonic fermentation media. The results indicate the capability of the tested nutraceutical formulations to beneficially modulate the composition and metabolite production of human intestinal microbiota and increase the antioxidant capacity in the intestinal environment.

Natural Products from Annonaceae as Potential Antichagasic Agents.

Chagas disease, a neglected tropical disease, is endemic in 21 Latin American countries and particularly prevalent in Brazil. Chagas disease has drawn more attention in recent years due to its expansion into non-endemic areas. The aim of this work was to computationally identify and experimentally validate the natural products from an Annonaceae family as antichagasic agents. Through the ligand-based virtual screening, we identified 57 molecules with potential activity against the epimastigote form of T. cruzi. Then, 16 molecules were analyzed in the in vitro study, of which, six molecules displayed previously unknown antiepimastigote activity. We also evaluated these six molecules for trypanocidal activity. We observed that all six molecules have potential activity against the amastigote form, but no molecules were active against the trypomastigote form. 13-Epicupressic acid seems to be the most promising, as it was predicted as an active compound in the in silico study against the amastigote form of T. cruzi, in addition to having in vitro activity against the epimastigote form.

In vitro inhibition of a multispecies oral cavity biofilm by Syzygium aromaticum essential oil.

OBJECTIVE: To evaluate the anti-fungal activity of Syzygium aromaticum essential oil and its inhibition of a multispecies biofilm in patients with oral candidiasis. BACKGROUND: Inhibiting biofilm formation on the denture surface is an important practice for preventing denture stomatitis. MATERIALS AND METHODS: The anti-fungal activity against Candida albicans and non-albicans Candida species was evaluated through the microdilution method to define Minimal Inhibitory (MIC) and Fungicidal (MFC) Concentrations. Time-kill assay assessed growth kinetics of C. albicans based on pre-determined time points (0, 1, 2, 4, 6 and 24 hours). A multi-species biofilm was formed using human saliva from patients with oral candidiasis and anti-biofilm activity determined by Colony Forming Units per milliliter (CFU/mL) count, fluorescence microscopy with calcofluor white to observe yeast presence and structure, and metabolic activity by XTT (2,3-Bis-(2Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-carboxanilide) reduction assay. RESULTS: The essential oil showed an anti-fungal activity against all Candida species (MIC 500-1000 µg/mL, MFC 1000-2000 µg/mL), and the time-kill assay showed that 2000 µg/mL (from 2 hours onward) and 1000 µg/mL (from 4 hours onward) concentrations had substantially lower yeast growth than the negative control. In the biofilm analysis, the essential oil had a lower CFU/mL count and a biofilm metabolic activity (91.4%) than seen with its negative control, and in both analyses, the essential oil was not significantly different from the positive control (chlorhexidine). Morphological analysis showed amorphous and fragmented cellular structures after treatment with the essential oil. CONCLUSION: Syzygium aromaticum essential oil had anti-fungal activities, reduced the Candida growth kinetics substantially and inhibited the multi-species biofilm formation.

Seasonal Variations of Polyphenols Content, Sun Protection Factor and Antioxidant Activity of Two Lamiaceae Species.

Secondary metabolite production by plants is influenced by external environmental factors that can change depending on the seasons, which makes it important to know how the plant, through its metabolism, is able to adapt to these variations. Mentha x villosa and Plectranthus amboinicus present in their chemical composition polyphenols, and through previous studies, it has been seen that these two species present promising in vitro photoprotective activity. The aim of this study was to evaluate seasonal alterations in photoprotective and antioxidant activities and the influence of factors such as precipitation levels and sun radiation incidence. Thus, polyphenol quantification, cromatographics (HPLC-DAD) and multivariate (PCA) analyses of extracts of the two species through twelve months were done. It was observed that the best months for photoprotective and antioxidant activities were September for M. villosa and July for P. amboinicus (SPF = 14.79). It was possible to conclude that solar radiation more clearly influences the production of phenolics and the increase of SPF in M. villosa, in addition to favoring the antioxidant activity of the two species, while precipitation seems to have no influence.

IMXQB-80: A Quillaja brasiliensis saponin-based nanoadjuvant enhances Zika virus specific immune responses in mice.

Vaccine adjuvants are compounds that enhance/prolong the immune response to a co-administered antigen. Saponins have been widely used as adjuvants for many years in several vaccines - especially for intracellular pathogens - including the recent and somewhat revolutionary malaria and shingles vaccines. In view of the immunoadjuvant potential of Q. brasiliensis saponins, the present study aimed to characterize the QB-80 saponin-rich fraction and a nanoadjuvant prepared with QB-80 and lipids (IMXQB-80). In addition, the performance of such adjuvants was examined in experimental inactivated vaccines against Zika virus (ZIKV). Analysis of QB-80 by DI-ESI-ToF by negative ion electrospray revealed over 29 saponins that could be assigned to known structures existing in their congener Q. saponaria, including the well-studied QS-21 and QS-7. The QB-80 saponins were a micrOTOF able to self-assembly with lipids in ISCOM-like nanoparticles with diameters of approximately 43 nm, here named IMXQB-80. Toxicity assays revealed that QB-80 saponins did present some haemolytical and cytotoxic potentials; however, these were abrogated in IMXQB-80 nanoparticles. Regarding the adjuvant activity, QB-80 and IMXQB-80 significantly enhanced serum levels of anti-Zika virus IgG and subtypes (IgG1, IgG2b, IgG2c) as well as neutralized antibodies when compared to an unadjuvanted vaccine. Furthermore, the nanoadjuvant IMXQB-80 was as effective as QB-80 in stimulating immune responses, yet requiring fourfold less saponins to induce the equivalent stimuli, and with less toxicity. These findings reveal that the saponin fraction QB-80, and particularly the IMXQB-80 nanoadjuvant, are safe and capable of potentializing immune responses when used as adjuvants in experimental ZIKV vaccines.

Larvicide Activity on Aedes aegypti of Essential Oil Nanoemulsion from the Protium heptaphyllum Resin.

The aim of this work was to prepare a nanoemulsion containing the essential oil of Protium heptaphyllum resin and to evaluate the larvicidal activity and the residual larvicidal effect against Aedes aegypti. The essential oil was identified by gas chromatography coupled to a mass spectrometer, and the nanoemulsions were prepared using a low-energy method and characterized by photon correlation spectroscopy. The results indicated the major constituents as p-cimene (27.70%) and α-Pinene (22.31%). Nanoemulsions had kinetic stability and a monomodal distribution in a hydrophilic-lipophilic balance of 14 with particle diameters of 115.56 ± 1.68 nn and zeta potential of -29.63 ± 3.46 mV. The nanoemulsion showed larvicidal action with LC50 = 2.91 µg∙mL-1 and residual larvicidal effect for 72 h after application to A. aegypti larvae. Consequently, the nanobiotechnological product derived from the essential oil of P. heptaphyllum resin could be used against infectious disease vectors.

Psychopharmacological effects of riparin III from Aniba riparia (Nees) Mez. (Lauraceae) supported by metabolic approach and multivariate data analysis.

BACKGROUND: Currently there is a high prevalence of humor disorders such as anxiety and depression throughout the world, especially concerning advanced age patients. Aniba riparia (Nees) Mez. (Lauraceae), popular known as "louro", can be found from the Amazon through Guianas until the Andes. Previous studies have already reported the isolation of alkamide-type alkaloids such as riparin III (O-methyl-N-2,6-dyhydroxy-benzoyl tyramine) which has demonstrated anxiolytic and antidepressant-like effects in high doses by intraperitoneal administration. METHODS: Experimental protocol was conducted in order to analyze the anxiolytic-like effect of riparin III at lower doses by intravenous administration to Wistar rats (Rattus norvegicus) (n = 5). The experimental approach was designed to last 15 days, divided in 3 distinct periods of five days: control, anxiogenic and treatment periods. The anxiolytic-like effect was evaluated by experimental behavior tests such as open field and elevated plus-maze test, combined with urine metabolic footprint analysis. The urine was collected daily and analyzed by 1H NMR. Generated data were statistically treated by Principal Component Analysis in order to detect patterns among the distinct periods evaluated as well as biomarkers responsible for its distinction. RESULTS: It was observed on treatment group that cortisol, biomarker related to physiological stress was reduced, indicating anxiolytic-like effect of riparin III, probably through activation of 5-HT2A receptors, which was corroborated by behavioral tests. CONCLUSION: 1H NMR urine metabolic footprint combined with multivariate data analysis have demonstrated to be an important diagnostic tool to prove the anxiolytic-like effect of riparin III in a more efficient and pragmatic way.

Computer-Aided Chemotaxonomy and Bioprospecting Study of Diterpenes of the Lamiaceae Family.

Lamiaceae is one of the largest families of angiosperms and is classified into 12 subfamilies that are composed of 295 genera and 7775 species. It presents a variety of secondary metabolites such as diterpenes that are commonly found in their species, and some of them are known to be chemotaxonomic markers. The aim of this work was to construct a database of diterpenes and to use it to perform a chemotaxonomic analysis among the subfamilies of Lamiaceae, using molecular descriptors and self-organizing maps (SOMs). The 4115 different diterpenes corresponding to 6386 botanical occurrences, which are distributed in eight subfamilies, 66 genera, 639 different species and 4880 geographical locations, were added to SistematX. Molecular descriptors of diterpenes and their respective botanical occurrences were used to generate the SOMs. In all obtained maps, a match rate higher than 80% was observed, demonstrating a separation of the Lamiaceae subfamilies, corroborating with the morphological and molecular data proposed by Li et al. Therefore, through this chemotaxonomic study, we can predict the localization of a diterpene in a subfamily and assist in the search for secondary metabolites with specific structural characteristics, such as compounds with potential biological activity.

Alkaloids of the Lauraceae.

This chapter presents an overview of the chemistry and pharmacology of the alkaloids found in species of the Lauraceae family. The occurrence of alkaloids from Lauraceae species as well as their chemical structures is summarized in informative and easy-to-understand tables. Within the Lauraceae family, the genera Ocotea (195), Litsea (180), Cryptocarya (133), and Neolitsea (110) have led to the greater number of publications regarding alkaloids content. Valuable and comprehensive information about the structure of these alkaloids is provided. The alkaloids of the aporphine type, found in 22 of the 23 genera, represent the predominant group in this family. Many of the isolated alkaloids exhibit unique structures. From plants of this family, 22 different types of skeletons have been isolated, among them only the purine alkaloids are classified as pseudoalkaloids, and the types phenethylamines, phenethylcinnamides, and phthalidoisoquinoline are classified as protoalkaloids. The chapter is presented as a contribution for the scientific community, mainly to enable the search for alkaloids in species belonging to the Lauraceae family.

Fluorescent Canthin-6-one Alkaloids from Simaba bahiensis: Isolation, Identification, and Cell-Labeling Properties.

Canthin-6-one alkaloids, which are present in plants of the genus Simaba, are natural compounds that are capable of acting as fluorescent probes. However, the chemical composition and fluorescent properties of most species of this genus have not been analyzed. The objective of this study was to characterize the fluorescent properties of an extract of S. bahiensis and identify the chemical entities responsible for these properties. In addition, the cell-labeling properties of the fluorescent dye from A and of the isolated compounds were characterized by confocal fluorescence microscopy and flow cytometry. One quassinoid and three fluorescent alkaloids were isolated from S. bahiensis, all compounds were identified by using NMR spectroscopy and high-resolution mass spectrometry. Staining experiments and HPLC-FL analysis shown that canthin-6-one alkaloids are the main green fluorescent compounds in the analyzed dyes. All compounds evaluated showed a cytoplasmic marker with a residence time of 24 h. The present study is the first to describe the presence of canthin-6-one alkaloids in S. bahiensis, in addition to demonstrating promising cell-labeling properties of fluorescent compounds from S. bahiensis with broad emission wavelengths.

Evaluación fitoquímica de hojas de Excoecaria lucida Sw. (Aité) (Euphorbiaceae) y Aislamiento e Identificación de un hemiterpenoide / Fitochemistry evaluation of leaves of Excoecaria lucida Sw. (Aité) (Euphorbiaceae) and isolation e identification of a hemiterpenoid

Introducción: excoecaria lucida Sw. (Aité), es una especie que crece en Cuba y en la región del Caribe. La misma ha sido poco estudiada por científicos a pesar de que la población la utiliza con fines medicinales, en lo principal como antiasmático, antimicrobiano y para el dolor dental. Objetivos: determinar la composición química de las hojas, así como aislar e identificar estructuralmente metabolitos secundarios. Métodos: las hojas secas y molidas, se maceraron 72 horas con etanol al 95 por ciento y se repitió el procedimiento en 4 ocasiones. Al extracto total se le determinó su composición química cualitativa y posterior se fraccionó con solventes. La fase de mayor rendimiento fue estudiada por Cromatografía Líquida de Alta Resolución y aislada la sustancia mayoritaria, la cual fue caracterizada por espectroscopía infrarroja y de Resonancia Magnética Nuclear 1H y 13C a 500 MHz (HMQC, HMBC, COSY y NOESY). Resultados: se determinó la presencia cualitativa de alcaloides, triterpenos y esteroides, quinonas, flavonoides, cumarinas, lactonas sesquiterpénicas, fenoles y taninos. De la fase de mayor rendimiento (butanólica= 23,48 g) se logró aislar e identificar el compuesto mayoritario perteneciente a un hemiterpenoide. Conclusiones: los metabolitos secundarios detectados coinciden con los reportes para otras especies del género. Se informa por primera vez para el género y la especie el aislamiento e identificación del hemiterpenoide (2E)-2-metil-2-buten-1,4-diol-1-O-β-D-glucopiranósido(AU)

Introduction: excoecaria lucida Sw. (Aité), it is a species that grow in Cuba and at the Caribbean region. It has been scientifically quite a little studied, in spite of the fact that the population utilizes it due to their medicinal properties as antiasthmatic, antimicrobial, and for the toothache treatment. Objectives: to determine the chemical composition of leaves, as well as the isolation and identification of their secondary metabolites. Methods: dried and milled leaves were macerated 72 hours with ethanol 95 percent, repeating the procedure in 4 occasions. Total extract was determined in his chemical qualitative composition and afterwards it was fragmented with solvents. The phase with greater yield was studied by High Performance Liquid Chromatography and isolated the main substance, which was characterized by infrared and Nuclear Magnetic Resonance spectroscopy: uni-dimensional 1H and 13C at 500 MHz and bi-dimensional HMQC, HMBC, COSY and NOESY. Results: the identification of Alkaloids, triterpenes and steroids, quinone, flavonoids, coumarins, sesqiterpenic lactones, tannins and phenols were accomplished. From the greater yield phase (butanol= 23.48 g) was isolated and identified the main compound, a hemiterpenoid. Conclusions: the identified secondary metabolites are in coincidence with other reports related to the genera. For first time the hemiterpene (2E)-2-methyl-2- buten-1,4-di-hydroxyl-1-O-β-D-glucopyranoside is informed in this genera and specie(AU)

Evaluación fitoquímica de hojas de Excoecaria lucida Sw. (Aité) (Euphorbiaceae) y Aislamiento e Identificación de un hemiterpenoide / Fitochemistry evaluation of leaves of Excoecaria lucida Sw. (Aité) (Euphorbiaceae) and isolation e identification of a hemiterpenoid

INTRODUCCIÓN: excoecaria lucida Sw. (Aité), es una especie que crece en Cuba y en la región del Caribe. La misma ha sido poco estudiada por científicos a pesar de que la población la utiliza con fines medicinales, en lo principal como antiasmático, antimicrobiano y para el dolor dental. OBJETIVOS: determinar la composición química de las hojas, así como aislar e identificar estructuralmente metabolitos secundarios. MÉTODOS: las hojas secas y molidas, se maceraron 72 horas con etanol al 95 % y se repitió el procedimiento en 4 ocasiones. Al extracto total se le determinó su composición química cualitativa y posterior se fraccionó con solventes. La fase de mayor rendimiento fue estudiada por Cromatografía Líquida de Alta Resolución y aislada la sustancia mayoritaria, la cual fue caracterizada por espectroscopía infrarroja y de Resonancia Magnética Nuclear 1H y 13C a 500 MHz (HMQC, HMBC, COSY y NOESY). RESULTADOS: se determinó la presencia cualitativa de alcaloides, triterpenos y esteroides, quinonas, flavonoides, cumarinas, lactonas sesquiterpénicas, fenoles y taninos. De la fase de mayor rendimiento (butanólica= 23,48 g) se logró aislar e identificar el compuesto mayoritario perteneciente a un hemiterpenoide. CONCLUSIONES: los metabolitos secundarios detectados coinciden con los reportes para otras especies del género. Se informa por primera vez para el género y la especie el aislamiento e identificación del hemiterpenoide (2E)-2-metil-2-buten-1,4-diol-1-O-ß-D-glucopiranósido.

INTRODUCTION: excoecaria lucida Sw. (Aité), it is a species that grow in Cuba and at the Caribbean region. It has been scientifically quite a little studied, in spite of the fact that the population utilizes it due to their medicinal properties as antiasthmatic, antimicrobial, and for the toothache treatment. OBJECTIVES: to determine the chemical composition of leaves, as well as the isolation and identification of their secondary metabolites. METHODS: dried and milled leaves were macerated 72 hours with ethanol 95 %, repeating the procedure in 4 occasions. Total extract was determined in his chemical qualitative composition and afterwards it was fragmented with solvents. The phase with greater yield was studied by High Performance Liquid Chromatography and isolated the main substance, which was characterized by infrared and Nuclear Magnetic Resonance spectroscopy: uni-dimensional 1H and 13C at 500 MHz and bi-dimensional HMQC, HMBC, COSY and NOESY. RESULTS: the identification of Alkaloids, triterpenes and steroids, quinone, flavonoids, coumarins, sesqiterpenic lactones, tannins and phenols were accomplished. From the greater yield phase (butanol= 23.48 g) was isolated and identified the main compound, a hemiterpenoid. CONCLUSIONS: the identified secondary metabolites are in coincidence with other reports related to the genera. For first time the hemiterpene (2E)-2-methyl-2- buten-1,4-di-hydroxyl-1-O-ß-D-glucopyranoside is informed in this genera and specie.

Antinociceptive activity of the extract of Erythroxylum caatingae / Actividad antinociceptiva del extracto de Erythroxylum caatingae

The genus Erythoxylum plants are popularly used as anti-inflammatory, anti-bacterial, and diuretic agentes, and for treating of respiratory problems. This study investigated the antinociceptive activity of methanolic extract of Erythroxylum caatingae (EcME) in chemical (formalin test and acetic acid test) and thermal models (hot plate test) of nociception. Intraperitoneal pre-treatment with EcME reduced the number of abdominal contortions and the licking time in the second phase of the formalin test. EcME did not show a significant effect in the hot plate test, rota-rod test, and the elevated plus maze test. These findings indicate that the antinociceptive activity of EcME is not because of a depressor effect on the central nervous system, and EcME is not a muscle relaxant. Nevertheless, Erythroxylum caatingae demonstrated peripheral antinociceptive activity, which confirms its popular use and contributes to the scientific knowledge of the species.

El género Erythoxylum es popularmente utilizado como agente anti-inflamatorio, antibacteriano y para el tratamiento de problemas respiratorios. Este estudio tiene como objetivo investigar la actividad antinociceptiva del extracto metanólico Erythroxylum caatingae (EcME), utilizando modelos químicos (prueba de la formalina y prueba de ácido acético) y térmico (prueba de la placa caliente) de nocicepción. El pretratamiento EcME por la vía intraperitoneal (i.p.), fue capaz de reducir el número de contorsiones abdominales y el tiempo de lamida en la segunda fase del test de formalina. EcME no tuvo efecto significativo en el test de la placa caliente, Rota-Rod y Laberinto en Cruz Elevado, mostrando que el efecto antinociceptivo no está relacionado con un efecto depresor del sistema nervioso central o miorelajante. Los datos experimentales muestran que Erythroxylum caatingae posee una actividad antinociceptiva periférica que confirma su uso popular, contribuyendo para el conocimiento científico de la especie.

Structural elucidation and NMR assignments of a new pyrrolizidine alkaloid from Crotalaria vitellina Ker Gawl.

A new pyrrolizidine alkaloid, named crotavitelin, was isolated from fruits of Crotalaria vitellina, Fabaceae (Papilionoideae). The structure was established by spectroscopic techniques such as one-dimensional and two-dimensional NMR, IR, and MS.