Mitracarpus frigidus reduces lipid metabolism and PGE2 levels in inflammatory cells.

OBJECTIVES: To evaluate the ability of the aqueous extract of Mitracarpus frigidus (MFAq) to inhibit lipid body formation and inflammatory mediator production in macrophages stimulated with lipopolysaccharide (LPS) and interferon gamma (IFN-γ). METHODS: MFAq was chemically characterized by ultrafast liquid chromatography/quadruple time-of-flight tandem mass spectrometry. The macrophages obtained from mice were incubated with MFAq. Cell viability and membrane integrity were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and propidium iodide assays, respectively. Moreover, NO, reactive oxygen species (ROS), transforming growth factor beta (TGF-ß), prostaglandin E2 (PGE2) levels and lipid bodies (LBs) were examined in macrophages that were stimulated with LPS and IFN-γ and treated with MFAq. Finally, molecular docking analysis was conducted to investigate the interaction of MFAq with the cyclooxygenase 2 (COX-2) enzyme. KEY FINDINGS: Chlorogenic acid, clarinoside, harounoside, rutin, kaempferol-3O-rutinoside and 2-azaanthraquinone were identified in MFAq. MFAq significantly inhibited NO, ROS and LBs, and did not affect the membrane integrity of macrophages. MFAq-treated cells showed significantly lower levels of TGF-ß and PGE2. Molecular docking demonstrated that the compounds found in MFAq are able to inhibit COX-2 by binding to important residues in the catalytic site. CONCLUSIONS: MFAq interferes with lipid metabolism in stimulated macrophages, leading to the reduction of important inflammatory mediators. Furthermore, MFAq can directly inhibit the COX-2 enzyme or inhibit its expression owing to its ability to reduce NO production.

Antifungal Annona muricata L. (soursop) extract targets the cell envelope of multi-drug resistant Candida albicans.

ETNOPHARMACOLOGICAL RELEVANCE: Annona muricata L. (soursop) is traditionally used in the treatment of inflammatory diseases, cancer, and infections caused by fungi. The therapeutic activity explored by its medicinal use is generally associated with its phytoconstituents, such as acetogenins and alkaloids. However, its potential antifungal bioactivity as well as its mechanism of action remains to be established. AIM OF THE STUDY: To evaluate the antifungal activity of the ethanolic extract of A. muricata leaves against multidrug-resistant Candida albicans (ATCC® 10231). MATERIAL AND METHODS: Phytoconstituents were detected by UFLC-QTOF-MS. The minimum inhibitory concentration was determined, followed by the determination of the minimum fungicidal concentration. For planktonic cells, the growth curve and cell density were evaluated. Studies to understand the mechanism of action on the cell envelope involved crystal violet permeability, membrane extravasation, sorbitol protection, exogenous ergosterol binding assay, metabolic activity, and cell viability. Furthermore, mitochondrial membrane potential was assessed. RESULTS: Our analyses demonstrated a significant inhibitory effect of A. muricata, with the ability to reduce fungal growth by 58% and cell density by 65%. The extract affected both the fungal plasma membrane and cell wall integrity, with significant reduction of the cell viability. Depolarization of the fungal mitochondrial membrane was observed after treatment with A. muricata. Rutin, xi-anomuricine, kaempferol-3O-rutinoside, nornuciferine, xylopine, atherosperminine, caffeic acid, asimilobine, s-norcorydine, loliolide, annohexocin, annomuricin, annopentocin, and sucrose were identified as extract bioactive components. CONCLUSIONS: Our findings show that the A. muricata extract is a source of chemical diversity, which acts as a potential antifungal agent with promising application to the therapy of infections caused by C. albicans.

Isolation and Chemical Characterization of Antifungal, Antioxidant, and Anti-Inflammatory Compounds from Centrosema coriaceum using GC/MS, UFLC-QTOF-MS, and FACE.

In recent years, natural products with biological activities have been increasingly researched. The elucidation of phytoconstituents is necessary for the development of drugs as a natural alternative for the treatment of various diseases. The work aimed to evaluate in vitro and in silico bioactivities of hexane (CCHE) and methanol (CCME) fractions of ethanolic extract from Centrosema coriaceum Benth (Fabaceae) leaves and elucidate their phytoconstituents. CCHE and CCME showed antifungal activity for Candida glabrata (MIC of 1000 µg/mL) with fungistatic effect and action in cell envelope by sorbitol and ergosterol assays. CCHE and CCME presented promising antioxidant activity against the DPPH radical with IC50 of 13.61±0.50 and 6.31±0.40 µg/mL, respectively, and relative antioxidant activity (RAA%) of 45.77±3.61/ 28.53±2.25 % for CCHE and 82.18±2.25/51.99±3.23 % for CCME when compared to rutin and quercetin, respectively. Moreover, these fractions demonstrated promising results for the inhibition of lipid peroxidation by ß-carotene/linoleic acid assay. For anti-inflammatory and cytotoxicity activities, CCHE and CCME significantly inhibited the production of nitric oxide and TNF-α, without toxicity on murine intraperitoneal macrophages, respectively. Esters, alkanes, steroids, tocopherols, and terpenes were identified in CCHE by GC/MS. Flavonoids, phenolic acids, and disaccharides were detected in CCME by UFLC-QTOF-MS and FACE. Furthermore, rutin was purified from CCME. In silico predictions evidenced that compounds present in both fractions have high affinity to the fungal membrane besides antioxidant and anti-inflammatory activities. Based on these observations, CCHE and CCME have a noteworthy potential for the design of novel antifungal and anti-inflammatory agents that should be explored in future studies.

In vivo anti-inflammatory and antinociceptive effects, and in vitro antioxidant, antiglycant and anti-neuroinflammatory actions of Syzygium malaccense.

Syzygium malaccense is popularly used to treat inflammation and pain-related ailments. The species was assessed regarding its antioxidant, antiglycant, anti-inflammatory, including anti-neuroinflammatory, and antinociceptive activities. Different models were employed to measure S. malaccense extract (ESM) antioxidant activity. The antiglycant activity was determined using the glucose-induced protein glycation model. LPS-induced neuroinflammation on murine BV-2 microglial cell line was used for anti-neuroinflammatory activity evaluation. The croton oil-induced ear edema test was accomplished to evaluate the in vivo anti-inflammatory activity. Acetic acid-induced writhing together with formalin-induced paw licking assays were performed to evaluate the antinociceptive potential. Finally, the chemical characterization was accomplished by a UHPLC-MS analysis. ESM presented relevant antioxidant and antiglycant activity. NO production by BV-2 cells was reduced, indicating the relevant neuroprotective activity. ESM significantly decreased the mice ear edema induced by croton oil and the nociceptive stimulus induced by acetic acid and formalin by central and peripheral mechanisms. The flavonoids myricitrin, myricetin and quercetin were identified and, as far as we know, the alkaloid reserpine was reported in the species for the first time. The antioxidant and antiglycant potential of ESM, may be related to the in vivo anti-inflammatory and antinociceptive effects, and to the in vitro neuroinflammation inhibition.

Spilanthol as a promising antifungal alkylamide for the treatment of vulvovaginal candidiasis.

Spilanthol is a bioactive alkylamide from the native Amazon plant species, Acmella oleracea. However, antifungal activities of spilanthol and its application to the therapeutic treatment of candidiasis remain to be explored. This study sought to evaluate the in vitro and in vivo antifungal activity of spilanthol previously isolated from A. oleracea (spilanthol(AcO)) against Candida albicans ATCC® 10231™, a multidrug-resistant fungal strain. Microdilution methods were used to determine inhibitory and fungicidal concentrations of spilanthol(AcO). In planktonic cultures, the fungal growth kinetics, yeast cell metabolic activity, cell membrane permeability and cell wall integrity were investigated. The effect of spilanthol(AcO) on the proliferation and adhesion of fungal biofilms was evaluated by whole slide imaging and scanning electron microscopy. The biochemical composition of the biofilm matrix was also analyzed. In parallel, spilanthol(AcO) was tested in vivo in an experimental vulvovaginal candidiasis model. Our in vitro analyses in C. albicans planktonic cultures detected a significant inhibitory effect of spilanthol(AcO), which affects both yeast cell membrane and cell wall integrity, interfering with the fungus growth. C. albicans biofilm proliferation and adhesion, as well as, carbohydrates and DNA in biofilm matrix were reduced after spilanthol(AcO) treatment. Moreover, infected rats treated with spilanthol(AcO) showed consistent reduction of both fungal burden and inflammatory processes compared to the untreated animals. Altogether, our findings demonstrated that spilanthol(AcO) is an bioactive compound against planktonic and biofilm forms of a multidrug resistant C. albicans strain. Furthermore, spilanthol(AcO) can be potentially considered for therapeutical treatment of vulvovaginal candidiasis caused by C. albicans. LAY SUMMARY: This study sought to evaluate the antifungal activity of spilanthol against Candida albicans ATCC® 10 231™, a multidrug-resistant fungal strain. Our findings demonstrated that spilanthol(AcO) can be potentially considered for therapeutical treatment of vulvovaginal candidiasis caused by C. albicans.

The essential oil from the fruits of the Brazilian spice Xylopia sericea A. St.-Hil. presents expressive in-vitro antibacterial and antioxidant activity.

OBJECTIVES: The aims of this study were to investigate the chemical composition and the antioxidant activity and antibacterial activity of the essential oil of Xylopia sericea fruits (OXS). The fruits of this species are popularly used for medicinal purposes, and as a condiment in food preparation. METHODS: The chemical composition of OXS was analysed by GC/MS. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) scavenging, ß-carotene/linoleic acid bleaching and phosphomolybdenum and thiobarbituric acid-reactive substance (TBARS) assays were used to evaluate the antioxidant activity. Antibacterial activity was assessed by minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against bacterial strains of interest to human health and food spoilage. KEY FINDINGS: Eighty-four compounds were identified. The sesquiterpenes spathulenol (16.42%), guaiol (13.93%) and germacrene D (8.11%) were the most abundant constituents. OXS presented a significant antioxidant activity and also a high bacteriostatic effect against Staphylococcus aureus, Enterobacter cloacae, Bacillus cereus and Klebsiella pneumoniae. CONCLUSIONS: Those results evidenced the potential of OXS to treat human bacterial infections and as an antimicrobial ingredient for food preservation.

Anthropogenic impact on a protected area, Rio Doce Park / Impacto antropogénico en un área protegida, el Parque Rio Doce

This study aimed to assess the anthropogenic activities at Rio Doce Park, Minas Gerais, Brazil, from ethnopharmacological surveys in Timóteo and Marliéria, which are located around the park. Interviews were conducted with previously identified, key informants, 15 in Timóteo and 10 in Marliéria. Two respondents collected medicinal plants in the forest of the park (from a few trees) but kept the same vulnerability of the use of their barks. Among the 141 surveyed botanical terms, we identified 95 species of 44 different botanical families. On the basis of statistical analyses, the 12 most used species were selected by respondents considering their therapeutic purposes and also obtaining the purpose of the use and dosage, among others. The knowledge about the use of medicinal plants has been maintained through generations but away from the formal health system and a sustainable management plan to encourage the preservation of the park.

Este estudio tuvo como objetivo evaluar las actividades humanas en Rio Doce Park, MG, desde encuesta ethnopharmacological en las ciudades de Timoteo y Marliéria. Realizamos entrevistas semi-estructuradas con informantes clave identificados anteriormente, un total de 15 en Timoteo y 10 en Marliéria. Sólo dos encuestados informaron de recolección de plantas medicinales en el parque forestal. De los 141 términos botánicos citados, se identificaron 95 especies y 44 familias de plantas. Basado en el análisis estadístico, 12 especies fueron seleccionadas más utilizados por los encuestados y sus efectos terapéuticos, también la obtención de la finalidad del uso y la dosis, entre otros. Se encontró que se perpetúan los conocimientos combate el uso de plantas medicinales, pero desconectado del sistema formal de salud y un plan de manejo sostenible para promover la conservación del parque.

Antimicrobial, antioxidant and cytotoxicity potential of Manihot multifida (L.) Crantz (Euphorbiaceae).

Manihot multifida (L.) Crantz (Euphorbiaceae) is widely used in popular medicine for the treatment of infected wounds. This study evaluated the in vitro antioxidant and antimicrobial potential of this species against strains of Gram-positive and Gram-negative bacteria and fungi, known to cause infections in humans. The extracts showed minimal inhibitory concentration (MIC) varying from 39 to 2500 µg/mL for antimicrobial activity. The methanolic extract of fruits, aqueous and hexane extracts of leaves showed a very strong activity against Candida albicans (ATCC 18804) with MIC of 39 µg/mL. Furthermore, the methanolic extract of M. multifida leaves exhibited DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging potential with inhibitory concentration (IC50) values of 46.9 µg/mL, followed by hexane extract of leaves with IC50 values of 59.2 µg/mL. The cytotoxic activity against brine shrimp was stronger for the methanolic extract of leaves (lethal concentration - LC50 of 15.6 µg/mL). These results suggest that M. multifida has interesting antimicrobial and antioxidant activities. Moreover, these results corroborate the popular use of this specie in treating fungal infections since it demonstrates significant activity against C. albicans.

Synergy in ethnopharmacological data collection methods employed for communities adjacent to urban forest

Several ethnopharmacological techniques used for sampling and data collection cannot be used as they are for different areas and cultural groups. This study combined and adapted reported ethnopharmacological research techniques for sampling and data collection of medicinal plants in forests adjacent to urban areas, and evaluated their potential applicability. The areas considered in this study included the neighborhoods adjacent to the Botanical Garden of the Federal University of Juiz de Fora, MG, Brazil. Application of combined ethnopharmacological research techniques ensured a practical strategy and reliability in the collected data. Preliminary interviews with 303 questionnaires constituted the general sampling of the population, and interviews with eleven people knowledgeable of medicinal plants constituted specific sampling. Using the two techniques, it was possible to identify sixty species, which are stored in the CESJ Herbarium. Given the lack of specific ethnopharmacological research tools in the context of multi-cultural urban communities located adjacent to forest areas that require preservation, this study shows that the synergistic use of techniques provides more reliable and reproducible data. The combined use of these techniques provides safety and simplifies the tools for future use in similar studies.

Effects of light intensity on the distribution of anthocyanins in Kalanchoe brasiliensis Camb. and Kalanchoe pinnata (Lamk.) Pers.

This paper compares two medicinal species of Kalanchoe, which are often used interchangeably by the population, regarding the distribution of anthocyanins under the influence of four luminosity levels for 6 months. For the morphoanatomical analysis, the 6th stem node of each plant was sectioned. Usual histochemical tests revealed the presence of anthocyanins by cross sections of the stems, petioles and leaf blades. The petioles and leaf blades were submitted to the extraction with acidified methanol, and the anthocyanins were quantified by spectrophotometric readings. At the macroscopic level, it was noticed for both species a higher presence of anthocyanins in stems and petioles of plants under full sunlight. The microscopy of K. brasiliensis stems evidenced the deposition of anthocyanins in the subjacent tissue to the epidermis and cortex, which increased with light intensity. In K. pinnata a subepidermal collenchyma was observed, which interfered in the visualization of anthocyanins. In petioles and leaf blades of K. brasiliensis the deposition of anthocyanins was peripheral, and in K. pinnata it was also throughout the cortex. The quantification of anthocyanins in petioles showed in 70% of light higher averages than in 25%, but in leaf blades there were no significant results. This study contributes to the pharmacognosy of Kalanchoe and it is sustained by the description of flavonoids as biological markers of the genus.

Chemical and agronomic development of Kalanchoe brasiliensis Camb. and Kalanchoe pinnata Pers under light and temperature levels.

This study compares the development of Kalanchoe brasiliensis and Kalanchoe pinnata, which are medicinal species known as "saião" and "folha da fortuna" that are used interchangeably by the population for medicinal purposes. The experiment consisted of 20 plots/species planted in plastic bags with homogeneous substrate in a randomized design, which grown under light levels (25%, 50%, 70%, full sunlight) at environment temperature, and a treatment under a plastic with greater temperature range than the external environment. It was obtained for K. pinnata a greater plant height, total length of sprouts, stems, production and dry matter content of leaves than that obtained for K. brasiliensis, which achieved higher averages only for the length of lateral branches. The species showed increases in height, which varied in inverse proportion to the light, and it was observed the influence of temperature in K. pinnata. The production and dry matter content of leaves were proportional to the luminosity; the same occurred in the thickness of leaves for K. brasiliensis. In the swelling index and Brix degree, K. brasiliensis showed higher averages than K. pinnata. In relation to the total content of flavonoids it was not observed significant differences for both species. The analyzed parameters showed the main differences in the agronomic development of the two species.

Chemical and agronomic development of Kalanchoe brasiliensis Camb. and Kalanchoe pinnata (Lamk.) Pers under light and temperature levels

This study compares the development of Kalanchoe brasiliensis and Kalanchoe pinnata, which are medicinal species known as "saião" and "folha da fortuna" that are used interchangeably by the population for medicinal purposes. The experiment consisted of 20 plots/species planted in plastic bags with homogeneous substrate in a randomized design, which grown under light levels (25 percent, 50 percent, 70 percent, full sunlight) at environment temperature, and a treatment under a plastic with greater temperature range than the external environment. It was obtained for K. pinnata a greater plant height, total length of sprouts, stems, production and dry matter content of leaves than that obtained for K. brasiliensis, which achieved higher averages only for the length of lateral branches. The species showed increases in height, which varied in inverse proportion to the light, and it was observed the influence of temperature in K. pinnata. The production and dry matter content of leaves were proportional to the luminosity; the same occurred in the thickness of leaves for K. brasiliensis. In the swelling index and Brix degree, K. brasiliensis showed higher averages than K. pinnata. In relation to the total content of flavonoids it was not observed significant differences for both species. The analyzed parameters showed the main differences in the agronomic development of the two species.

Este estudo compara o desenvolvimento de Kalanchoe brasiliensis e Kalanchoe pinnata, espécies medicinais conhecidas como "saião" e "folha da fortuna" que são utilizadas indiferenciadamente pela população para fins medicinais. O experimento consistiu em 20 parcelas/espécie plantadas em sacos plásticos com substrato homogêneo, em delineamento ao acaso, cultivadas sob níveis de luminosidade (25 por cento, 50 por cento, 70 por cento, luz plena) em temperatura ambiente, e um tratamento sob plástico com maior amplitude térmica que o meio externo. Obteve-se para K. pinnata maiores alturas de plantas, comprimentos totais de brotos, caules, produção e teor de matéria seca de folhas que para K. brasiliensis, que alcançou maiores médias apenas para comprimentos de ramos laterais. As espécies apresentaram aumento em altura que variou em proporção inversa à incidência luminosa, e observou-se a influência da temperatura em K. pinnata. A produção e teor de matéria seca de folhas foram proporcionais à luminosidade; o que também ocorreu na espessura foliar para K. brasiliensis. No índice de intumescência e grau Brix, K. brasiliensis apresentou maiores médias que K. pinnata. Em relação ao teor de flavonoides totais, não foram observadas diferenças significativas para ambas as espécies. Os parâmetros analisados apontaram as principais diferenças no desenvolvimento agronômico das duas espécies.

Anti-allergic properties of the bromeliaceae Nidularium procerum: inhibition of eosinophil activation and influx.

New therapeutic approaches for the treatment of allergic diseases can be aided by the development of agents capable of regulating eosinophilic leukocytes. Here, we evaluated the anti-allergic properties of a crude extract of the Brazilian bromeliaceae Nidularium procerum, focusing on its effects on allergic eosinophilia. By studying allergic pleurisy in actively sensitized C57Bl/6 mice, we observed that pretreatment with N. procerum (2 mg/kg; i.p.) reduced pleural eosinophil influx triggered by allergen challenge. N. procerum was also able to reduce lipid body numbers found within infiltrating eosinophils, indicating that N. procerum in vivo is able to affect both migration and activation of eosinophils. Consistently, pretreatment with N. procerum blocked pleural eosinophil influx triggered by PAF or eotaxin, key mediators of the development of allergic pleural eosinophilia. The effect of N. procerum was not restricted to eosinophils, since N. procerum also inhibited pleural neutrophil and mononuclear cell influx. Of note, N. procerum failed to alter the acute allergic reaction, characterized by mast cell degranulation, oedema, and cysteinyl leukotriene release. N. procerum also had direct effects on murine eosinophils, since it inhibited both PAF- and eotaxin-induced eosinophil chemotaxis on an in vitro chemotactic assay. Therefore, N. procerum may be a promising anti-allergic therapy, inasmuch as it presents potent anti-eosinophil activity.