Herbal-drug interaction through the pregnane X receptor / Interacción de medicamentos y hierbas a través del receptor de pregnanos X

The interaction potential of Cynara scolymus L., Mikania glomerata Spreng.,Rhamnus purshiana DC and Uncaria tomentosa (Willd. Ex Roem. & Schult.) with conventional drugs metabolized by the CYP3A4 metabolic route was tested in HeLa cell lines, using the in vitro model of the hPXR. The herbal medicines C. scolymus (1.5 mg/mL dry extract) did not affect the receptor. On the other hand, M. glomerata (5.5 mg/mL dry extract), R. purshiana (1.5 mg/mL dry extract), and U. tomentosa (2.0 mg/mL dry extract) showed to be hPXR agonist, suggesting a potential interaction with the conventional drugs metabolized by the same isoforms of the CYP superfamily. The results from this study contribute to the use safer and more effective of these herbal medicines.

Se evaluó el potencial de interacción de Cynara scolymus L., Mikania glomerata Spreng., Rhamnus purshiana DC y Uncaria tomentosa (Willd. Ex Roem. & Schult.) con fármacos convencionales metabolizados por la ruta metabólica CYP3A4 en líneas celulares HeLa, utilizando el modelo in vitro del hPXR. Las hierbas medicinales C. scolymus (1,5 mg/mL de extracto seco) no afectaron al receptor. Por otro lado, M. glomerata (5.5 mg/mL de extracto seco), R. purshiana (1.5 mg/mL de extracto seco) y U. tomentosa (2.0 mg/mL de extracto seco) mostraron ser agonistas de hPXR, lo que sugiere una potencial interacción con los fármacos convencionales metabolizados por las mismas isoformas de la superfamilia CYP. Los resultados de este estudio contribuyen a un uso más seguro y eficaz de estos medicamentos a base de hierbas medicinales.

Antiprotozoal compounds from Mikania periplocifolia (Asteraceae).

Chagas disease, African trypanosomiasis and Leishmaniasis are neglected parasitic diseases which affect millions of people worldwide. In a previous work, we report the antiprotozoal activity of the dichloromethane extract of Mikania periplocifolia Hook. & Arn. (Asteraceae). The aim of this work was to isolate and identify the bioactive compounds present in the extract. The fractionation of the dichloromethane extract has led to the isolation of the sesquiterpene lactone miscandenin and the flavonoid onopordin, together with the sesquiterpene lactones mikanolide, dihydromikanolide and deoxymikanolide, which have previously shown antiprotozoal activity. Miscandenin and onopordin were assayed in vitro against Trypanosoma cruzi, T. brucei and Leishmania braziliensis. Miscandenin was active against T. cruzi trypomastigotes and amastigotes with IC50 values of 9.1 and 7.7 µg/ml, respectively. This sesquiterpene lactone and the flavonoid onopordin showed activity against T. brucei trypomastigotes (IC50 = 0.16 and 0.37 µg/ml) and L. braziliensis promastigotes (IC50 = 0.6 and 1.2 µg/ml), respectively. The CC50 values on mammalian cells were 37.9 and 53.4 µg/ml for miscandenin and onopordin, respectively. Besides, the pharmacokinetic and physicochemical properties of miscandenin were assessed in silico, showing a good drug-likeness profile. Our results highlight this compound as a promising candidate for further preclinical studies in the search of new drugs for the treatment of trypanosomiasis and leishmaniasis.

Ent-kaurenoic acid-enriched Mikania glomerata leaves-complexed ß-cyclodextrin: Pharmaceutical development and in vivo antitumor activity in a sarcoma 180 mouse model.

The extract obtained from Mikania glomerata leaves rich in ent-kaurenoic acid (ERKA) shows cytotoxic activity in vitro, but its hydrophobic nature and thermosensitivity are issues to be solved prior to in vivo antitumor studies. The purpose of this study was to investigate the antitumor activity of inclusion complexes formed between ERKA and ß-cyclodextrin (ERKA:ß-CD) in rodents. ERKA:ß-CD complexes obtained by malaxation (MX) and co-evaporation (CE) methods were firstly characterized regarding their physical properties, encapsulation efficiency, and cytotoxicity againts L929 cells. The antitumor activity study was then performed in mice with sarcoma 180 treated with saline, 5-fluouracil (5FU) and ERKA:ß-CD at 30, 100 and 300 µg/kg. The weight, volume, percentage of inhibition growth, gross and pathological features and positivity for TUNEL, ki67, NFκB and NRF2 in the tumors were assessed. Serum lactate-dehydrogenase activity (LDH), white blood cells count (WBC) and both gross and pathological features of the liver, kidneys and spleen were also evaluated. The formation of the inclusion complexes was confirmed by thermal analysis and FTIR, and they were non-toxic for L929 cells. The MX provided a better complexation efficiency. ERKA:ß-CD300 promoted significant tumor growth inhibition, and attenuated the tumor mitotic activity and necrosis content, comparable to 5-fluorouracil. ERKA:ß-CD300 also increased TUNEL-detected cell death, reduced Ki67 and NF-kB immunoexpression, and partially inhibited the serum LDH activity. No side effect was observed in ERKA:ß-CD300-treated animals. The ERKA:ß-CD inclusion complexes at 300 µg/kg displays antitumour activity in mice with low systemic toxicity, likely due to inhibition on the NF-kB signaling pathway and LDH activity.

Growth, leaf gas exchange and mycorrhizal colonization of three medicinal species submitted to different irradiance levels / Crescimento, trocas gasosas foliares e colonização micorrizica de três espécies de plantas medicinais submetidas a diferentes níveis de irradiância

This study o evaluated growth, leaf gas exchange and arbuscular mycorrhizal fungi root colonization in three medicinal plant species under different irradiance intensities. Fridericia chica (Bonpl.) L.G.Lohmann, Mikania laevigata Sch.Bip. ex Baker and Varronia curassavica Jacq. were propagated by cutting and cultivated for 120 days in artificially shaded environments using black shade-type screens, obtaining four irradiance levels: 100%, 70%, 50% and 30%. The experimental design was completely randomized in a 3 x 4 factorial scheme (three plant species and four irradiation levels) with seven replicates. The three medicinal species showed higher liquid assimilation, mass growth and arbuscular mycorrhizal fungi root colonization rates when exposed to environments with 70% light availability. In relation to physiological responses, V. curassavica presented higher photosynthetic rate, stomatal conductance and transpiration when submitted to 70% irradiance, being able to be cultivated in more open environments with higher irradiation levels. Conversely F. chica and M. laevigata presented shade tolerance characteristics. At the initial growth phase, the results obtained can be used as indicators to recommend the ideal cultivation environment for these species in agroforestry systems.

O objetivo deste trabalho foi avaliar o crescimento, as trocas gasosas foliares e a colonização por fungos micorrízicos arbusculares em três espécies de plantas medicinais, sob diferentes intensidades de irradiância. Fridericia chica (Bonpl.) L.G. Ohmmann, Mikania laevigata Sch.Bip. ex Baker e Varronia curassavica Jacq. foram propagadas por estaquia e cultivadas por 120 dias em ambientes artificialmente sombreados, utilizando telas do tipo sombrite, em quatro níveis de irradiância, 100%, 70%, 50% e 30%. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 3 x 4 (três espécies de plantas e quatro níveis de irradiação) com sete repetições. As três espécies medicinais avaliadas apresentaram maiores taxas de assimilação líquida, crescimento em massa e colonização radicular por fungos micorrízicos arbusculares quando expostas a ambientes com 70% de disponibilidade de luz. Em relação às respostas fisiológicas, V. curassavica apresentou maior taxa fotossintética, condutância estomática e transpiração quando submetidas a 70% de irradiância, podendo ser cultivada em ambientes mais abertos e com maiores níveis de radiação. Por outro lado, F. chica e M. laevigata apresentaram características de plantas tolerantes à sombra. Os resultados obtidos na fase inicial de crescimento podem ser utilizados como um indicador para recomendar o ambiente de plantio dessas três espécies medicinais em sistemas agroflorestais.

Comparative analyses of Mikania (Asteraceae: Eupatorieae) plastomes and impact of data partitioning and inference methods on phylogenetic relationships.

We assembled new plastomes of 19 species of Mikania and of Ageratina fastigiata, Litothamnus nitidus, and Stevia collina, all belonging to tribe Eupatorieae (Asteraceae). We analyzed the structure and content of the assembled plastomes and used the newly generated sequences to infer phylogenetic relationships and study the effects of different data partitions and inference methods on the topologies. Most phylogenetic studies with plastomes ignore that processes like recombination and biparental inheritance can occur in this organelle, using the whole genome as a single locus. Our study sought to compare this approach with multispecies coalescent methods that assume that different parts of the genome evolve at different rates. We found that the overall gene content, structure, and orientation are very conserved in all plastomes of the studied species. As observed in other Asteraceae, the 22 plastomes assembled here contain two nested inversions in the LSC region. The plastomes show similar length and the same gene content. The two most variable regions within Mikania are rpl32-ndhF and rpl16-rps3, while the three genes with the highest percentage of variable sites are ycf1, rpoA, and psbT. We generated six phylogenetic trees using concatenated maximum likelihood and multispecies coalescent methods and three data partitions: coding and non-coding sequences and both combined. All trees strongly support that the sampled Mikania species form a monophyletic group, which is further subdivided into three clades. The internal relationships within each clade are sensitive to the data partitioning and inference methods employed. The trees resulting from concatenated analysis are more similar among each other than to the correspondent tree generated with the same data partition but a different method. The multispecies coalescent analysis indicate a high level of incongruence between species and gene trees. The lack of resolution and congruence among trees can be explained by the sparse sampling (~ 0.45% of the currently accepted species) and by the low number of informative characters present in the sequences. Our study sheds light into the impact of data partitioning and methods over phylogenetic resolution and brings relevant information for the study of Mikania diversity and evolution, as well as for the Asteraceae family as a whole.

El guaco: un agente vegetal utilizado en el Nuevo Reino de Granada contra los síntomas generados por afecciones del sistema músculo-esquelético / Guaco: A plant agent used in the New Kingdom of Granada against symptoms generated by disorders of the musculoskeletal system

RESUMEN Objetivo: Contextualizar una receta médica de comienzos del siglo XIX en el Nuevo Reino de Granada, en la cual se prescribe el guaco para disminuir síntomas generados por afecciones del sistema músculo-esquelético. De igual forma, analizar, en función de los conocimientos actuales, cómo actúa la fórmula sobre mecanismos fisiopatológicos de la enfermedad, explicando la reducción del dolor y las secuelas asociadas. Material y método: Búsqueda documental en el Archivo Histórico Cipriano Rodríguez Santamaría, de la Biblioteca Octavio Arizmendi Posada, de la Universidad de La Sabana. Se analizó el documento denominado Reumatismo. Posteriormente se realizó una revisión de la literatura entre 1999-2018, en las bases de datos ScienceDirect/ClinicalKey/Scielo. Conclusiones: Existe evidencia científica que podría explicar la efectividad del guaco, usado en el Nuevo Reino de Granada por sus propiedades antiinflamatorias y analgésicas aportadas por componentes como la cumarina y los flavonoides. Sin embargo, una descripción vaga en la posología del guaco, signos, síntomas y comorbilidades que no se mencionan en la receta, dificulta analizar la eficacia del tratamiento y cómo lograba disminuir o controlar específicamente los síntomas dados por afecciones del sistema musculoesquelético con su aplicación. Esta tradición, en consecuencia, carece de sustento propiamente científico para el tratamiento médico de enfermedades osteomusculares.

ABSTRACT Objective: To contextualize a medical prescription of the early 19th century in the New Kingdom of Granada, in which guaco was prescribed to reduce symptoms caused by musculoskeletal system disorders, which were ill-defined at the time. Similarly, based on current knowledge, to analyse the manner in which the formula acts on pathophysiological mechanisms of rheumatic diseases, in order to explain the reduction of pain, and associated sequelae. Material and method: Documentary research into the Cipriano Rodríguez Santamaría Historical Archive of the Octavio Arizmendi Library of the University of La Sabana, in Chía, Colombia. The document analysed was called Rheumatism. Subsequently, a review of the literature was carried out in Science Direct / Clinical Key / Scielo databases in the period from 1999 to 2018. Conclusions: There is scientific evidence that supports the efficiency of guaco used in the Kingdom of New Granada due its anti-inflammatory and analgesic properties. However, a vague description of the dosage of the guaco, signs, symptoms, and comorbidities, which are not mentioned in the prescription, hinders the understanding of its application and the thorough effectiveness of the treatment in order to control the symptoms of musculoskeletal system conditions. This tradition, consequently, lacks proper scientific support for the medical treatment of musculoskeletal disorders.

GC-MS-FID characterization and antibacterial activity of the Mikania cordifolia essential oil and limonene against MDR strains.

The present study evaluated the effect of the essential oil of Mikania cordifolia (EOMc) and its major constituent limonene alone or associated with antibacterial drugs against Multidrug Resistant Bacteria (MDR). To evaluate the antibacterial activity, the minimum inhibitory concentrations (MIC) of the oil and limonene against Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were determined. The antibiotic-modulating activity was assessed using subinhibitory concentrations (MIC/8) of these substances in combination with conventional antibacterial drugs. Although no relevant antibacterial activity of the natural products was detected, both substances modulated the action of antibiotics against resistant bacteria. The EOMc demonstrated the best modulating effect against P. aeruginosa, presenting synergistic effects when associated with gentamicin and norfloxacin. In addition, the oil reduced the MIC of norfloxacin against E. coli as well as reduced the MIC of gentamicin against S. aureus. On the other hand, the best effect of limonene was obtained against S. aureus. Thus, it is concluded that the essential oil Mikania cordifolia and the isolated compound limonene do not have clinically significant antibacterial effect, but modulate the action of antibiotics against MDR bacteria.

Stability of hydroalcoholic extracts of two species of guaco; Mikania glomerata SPRENG. and Mikania laevigata SCHULTZ. (Asteraceae), by UHPLC-MS

It is important to study the stability of plant extracts used as active ingredients in phytotherapic medicine, as degradation of the active principles directly affects the efficacy and safety of these products. Therefore, a stability study of the hydroalcoholic extract of the species: Mikania glomerata and Mikania laevigata was conducted in order to determine the speed of degradation and shelf life of these extracts, which are incorporated in cough syrup in Brazil. Leaves of both species were dried in an oven or by lyophilization (freeze-dried). Hydroalcoholic extracts underwent both accelerated stability study of six months and long-term stability study for 12 months. Samples were stored at different temperatures and every three months were analysed by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) to monitor their chemical profile, quantifying coumarin and chlorogenic acid. For all conditions of the study, a reduction of the content of the chemical marker of this species, coumarin, greater than 5% was observed, so a shelf life of two years cannot be assigned to the hydroalcoholic extracts of these species as observed in commercial extracts.

Evaluation of genotoxicity and coumarin production in conventional and organic cultivation systems of Mikania glomerata Spreng.

Mikania glomerata Sprengel, popularly known as "guaco," is used in Brazilian folk medicine for several inflammatory and allergic conditions. Besides, the popular use "guaco" is indicated by the Brazilian Ministry of Health as a safe and effective herbal medicine. The biological activity of M. glomerata extracts is due to the presence of the coumarins, a large family of phenolic substances found in plants and is made of fused benzene and α-pyrone rings. Considering that there are few data on the biological effects of the extracts of M. glomerata, mainly in genetic level, this work aims to evaluate, in vitro, the genotoxicity and coumarin production in M. glomerata in conventional and organic growing. The data showed that the organic culture system showed double the concentration of coumarin being significantly more productive than the conventional system. Besides, the results of comet assay suggest that extracts of M. glomerata cultivated in a conventional system was genotoxic, increased DNA damage levels while the organic extracts seem to have antigenotoxic effect possibly due to the concentration of coumarins. Additional biochemical investigations are necessary to elucidate the mechanisms of action of M. glomerata extracts, which were found to have a role in protection against DNA damage.

Influence of environmental factors on the volatile composition of two Brazilian medicinal plants: Mikania laevigata and Mikania glomerata.

INTRODUCTION: Mikania laevigata Sch.Bip. ex Baker and Mikania glomerata Spreng. are medicinal plants popularly known as guaco, used for inflammatory diseases of the respiratory system, included in pharmaceutical formularies and often used without distinction. However, several studies show that the chemical composition varies between these species, as well as in plants are subjected to different environmental stresses. Few studies have been carried out with the volatile compounds of guaco, even less about the changes in volatile composition due to abiotic variation. OBJECTIVE: The aim of this work was to evaluate how volatile compounds vary according to the seasons and at different times of the day and if these compounds are influenced by the variations in the growth conditions such as: temperature, luminosity and water. METHODS: The headspace volatiles of the leaves were sampled by solid phase micro extraction and analyzed by gas chromatography-mass spectrometry. Untargeted metabolomic analysis of the resulting chromatograms and chemometrics was applied. The chemical profile of the volatiles of M. laevigata and M. glomerata were different; being clearly separated in the exploratory grouping analyzes (PCA), followed by analysis of variance of the marker compounds of both species. RESULTS: Only M. laevigata contained coumarin, considered to be the chemical marker of both species and to be responsible for the therapeutic activities. There was no significant difference between the morning and afternoon collections of either species. Coumarin, α-pinene and bicyclogermacrene were more intense in the volatiles of M. laevigata throughout the year and responsible for grouping the samples of this species. For M. glomerata, hexanal and 2-hexenal were responsible for grouping the samples and were more intense in all months. The growth conditions tested affected the intensity of specific compounds in the chromatograms. Some compounds were less intense with the increase of the temperature and in the plants subjected to full sunlight. However, certain volatile compounds-such as pinenes-were more intense in plants suffering drought. CONCLUSION: The variation in composition between species of guaco was greater than those observed in the seasonal and cultivation studies, indicating that these species cannot be used indistinctly.

Primary targets of the sesquiterpene lactone deoxymikanolide on Trypanosoma cruzi.

BACKGROUND: Deoxymikanolide is a sesquiterpene lactone isolated from Mikania micrantha and M. variifolia which, has previously demonstrated in vitro activity on Trypanosoma cruzi and in vivo activity on an infected mouse model. PURPOSE: Based on these promising findings, the aim of this study was to investigate the mechanism of action of this compound on different parasite targets. METHODS: The interaction of deoxymikanolide with hemin was examined under reducing and non- reducing conditions by measuring modifications in the Soret absorption band of hemin; the thiol interaction was determined spectrophotometrically through its reaction with 5,5'-dithiobis-2-nitrobenzoate in the presence of glutathione; activity on the parasite antioxidant system was evaluated by measuring the activity of the superoxide dismutase and trypanothione reductase enzymes, together with the intracellular oxidative state by flow cytometry. Superoxide dismutase and trypanothione reductase activities were spectrophotometrically tested. Cell viability, phosphatidylserine exposure and mitochondrial membrane potential were assessed by means of propidium iodide, annexin-V and rhodamine 123 staining, respectively; sterols were qualitatively and quantitatively tested by TLC; ultrastructural changes were analyzed by transmission electron microscopy. Autophagic cells were detected by staining with monodansylcadaverine. RESULTS: Deoxymikanolide decreased the number of reduced thiol groups within the parasites, which led to their subsequent vulnerability to oxidative stress. Treatment of the parasites with the compound produced a depolarization of the mitochondrial membrane even though the plasma membrane permeabilization was not affected. Deoxymikanolide did not affect the intracellular redox state and so the mitochondrial dysfunction produced by this compound could not be attributed to ROS generation. The antioxidant defense system was affected by deoxymikanolide at twenty four hours of treatment, when both an increased oxidative stress and decreased activity of superoxide dismutase and trypanothione reductase (40 and 60% respectively) were observed. Both the oxidative stress and mitochondrial dysfunction induce parasite death by apoptosis and autophagy. CONCLUSION: Based on our results, deoxymikanolide would exert its anti-T cruzi activity as a strong thiol blocking agent and by producing mitochondrial dysfunction.

Seasonal changes and solvent effects on fractionated functional food component yields from Mikania laevigata leaves.

Statistical mixture design extraction and fractionation of Mikania plant samples were carried out to quantitatively study seasonal and solvent composition effects as well as their interactions on secondary metabolites. The mixture design consisted of ethanol, acetone, dichloromethane and chloroform solvents and their binary, ternary and quaternary mixtures. Yields were measured for the crude extract and its neutral, organic, basic, polar and fiber fractions obtained with each solvent composition from samples harvested during 2010. Two-way ANOVA found statistically significant seasonal and solvent effects for the crude extract and all the fractions except for the polar fraction solvent. The best solvent extractor depends the harvest season. High crude, organic and polar fraction yields in the summer are correlated with the coumarin UV-vis absorbance at 274 nm. Crude yields of mixtures containing ethanol are correlated with the coumarin absorbances in summer, winter and spring whereas mixtures without ethanol show no significant correlation.

Radiation levels of UV-A and UV-B on growth parameters and coumarin content in guaco / Níveis de radiação UVA e UVB sobre parâmetros de crescimento e conteúdo de cumarina em guaco

Guaco is a medicinal plant species containing coumarin, a secondary metabolite whose concentration is influenced by various factors, such as the level of exposure to UVA (315-400 nm) and UVB (280-315 nm) radiation, age of the plant, and position of the storage organ in the plant. This study aimed to assess the effect of different blocking levels of UVA and UVB radiation on growth parameters and coumarin content of two guaco accessions. Treatments did not affect any of the growth parameters assessed, except leaf area. Coumarin content was higher in leaves collected from the upper canopy and with unblocked exposure to UVA and UVB.(AU)

Guaco é uma espécie medicinal que apresenta em sua composição a cumarina, um metabólito secundário cuja concentração no vegetal é influenciada por fatores como os níveis de radiação UV-A (315-400 nm) e UV-B (280-315 nm) incidentes, bem como pela idade e posição do órgão de armazenamento na planta. O objetivo deste estudo foi o de avaliar o efeito de diferentes níveis de bloqueio das radiações UV-A e UV-B sobre parâmetros de crescimento e teor de cumarina em dois acessos de guaco. Os tratamentos não apresentaram efeito sobre os parâmetros de crescimento avaliados, com exceção dos valores de área foliar. Em relação aos teores de cumarina, estes foram superiores nas folhas coletadas do dossel superior, bem como no tratamento sem bloqueio das radiações UV-A e UV-B incidentes.(AU)

Kaurenoic acid and its sodium salt derivative: antibacterial activity against Porphyromonas gingivalis and their mechanism of action.

AIM: To evaluate the antibacterial activity of 12 kaurane-type diterpenes against a panel of bacteria that cause endodontic infection. METHODS & MATERIALS: We conducted tests against bacteria in the planktonic or in the sessile mode, cytotoxic assays for the most promising compounds against human normal lung fibroblast cells, and Porphyromonas gingivalis (ATCC 33277) proteomic analysis. RESULTS & CONCLUSION: Kaurenoic acid and its salt exhibited satisfactory antibacterial action against the evaluated bacteria. Proteomic analysis suggested that these compounds might interfere in bacterial metabolism and virulence factor expression. Kaurane-type diterpenes are an important class of natural products and should be considered in the search for new irrigating solutions to treat endodontic infections.

Comparison of the Morphology, Anatomy, and Chemical Profile of Mikania glomerata and Mikania laevigata.

Mikania glomerata and Mikania laevigata are medicinal plants popularly used in Brazil for colds and asthma. Although several studies report differences in their composition, they are frequently used indistinctly, which may be due to misidentification of the species or the plant part used. Herein, the chemical, anatomical, and morphological profiles of both species were evaluated, seeking to differentiate them. Due to the pattern of secondary metabolites found in M. glomerata and M. laevigata, the chemical profile of the leaves was the characteristic that best differentiated the two species. Coumarin was present in all the leaf samples of M. laevigata but absent or in low concentration in leaf samples of M. glomerata. The stems presented coumarin in both species, albeit in different concentrations. The quantification of chlorogenic and dicaffeoylquinic acids was crucial for the separation of the two species, as higher concentrations are found in M. glomerata. The anatomic structure of the leaves, stem, and petioles of both species was conserved, even when exposed to different environments, but presented only subtle differences between species. The morphology of the leaves presented different characteristics for each species but altered when exposed to shade. Therefore, the chemical analysis together with the morphological characteristics of plants grown in full sunlight may be used for the correct identification of these species.

Decontamination of Mikania glomerata Leaves by Gamma Irradiation: Coumarin Determination by HPLC-DAD, Microbiological Control and Genotoxicological Studies.

Gamma irradiation as a decontaminating physical agent could be an important tool in the production chain of herbal medicines by improving the microbiological quality of raw materials and the safety of final products. This study was undertaken to investigate the genotoxic potential and eventual chemical modifications of a batch of Mikania glomerata raw material decontaminated by different doses of gamma irradiation (2.0, 3.5, and 5.0 kGy), using a cesium-137 source. DNA damage was assessed in vitro by agarose gel electrophoresis in regard to double-chain breaks of plasmid pUC 9.1 DNA and in vivo by micronucleus test in bone marrow cells of Wistar rats. Cytotoxicity in bone marrows was assessed by scoring polychromatic and normochromatic erythrocytes ratio. An HPLC-DAD method was adapted and validated for the enhancement of coumarin selectivity from the other matrix constituents. The microbial load was satisfactorily reduced, leading to sterilization at the highest dose. Genotoxic and cytotoxic effects were not increased in the in vitro and in vivo models. The concentration of coumarin and the chromatographic profiles of the hydroalcoholic plant extracts (ethanol 70% v/v) were not modified after such process. Therefore, this work suggests that gamma irradiation of M. glomerata raw material is suitable and safe for microbiological control purposes at the employed doses.

Assessment of sesquiterpene lactones isolated from Mikania plants species for their potential efficacy against Trypanosoma cruzi and Leishmania sp.

Four sesquiterpene lactones, mikanolide, deoxymikanolide, dihydromikanolide and scandenolide, were isolated by a bioassay-guided fractionation of Mikania variifolia and Mikania micrantha dichloromethane extracts. Mikanolide and deoxymikanolide were the major compounds in both extracts (2.2% and 0.4% for Mikania variifolia and 21.0% and 6.4% for Mikania micrantha respectively, calculated on extract dry weight). Mikanolide, deoxymikanolide and dihydromikanolide were active against Trypanosoma cruzi epimastigotes (50% inhibitory concentrations of 0.7, 0.08 and 2.5 µg/mL, for each compound respectively). These sesquiterpene lactones were also active against the bloodstream trypomastigotes (50% inhibitory concentrations for each compound were 2.1, 1.5 and 0.3 µg/mL, respectively) and against amastigotes (50% inhibitory concentrations for each compound were 4.5, 6.3 and 8.5 µg/mL, respectively). By contrast, scandenolide was not active on Trypanosoma cruzi. Besides, mikanolide and deoxymikanolide were also active on Leishmania braziliensis promastigotes (50% inhibitory concentrations of 5.1 and 11.5 µg/mL, respectively). The four sesquiterpene lactones were tested for their cytotoxicity on THP 1 cells. Deoxymikanolide presented the highest selectivity index for trypomastigotes (SI = 54) and amastigotes (SI = 12.5). In an in vivo model of Trypanosoma cruzi infection, deoxymikanolide was able to decrease the parasitemia and the weight loss associated to the acute phase of the parasite infection. More importantly, while 100% of control mice died by day 22 after receiving a lethal T. cruzi infection, 70% of deoxymikanolide-treated mice survived. We also observed that this compound increased TNF-α and IL-12 production by macrophages, which could contribute to control T. cruzi infection.

Mikania glomerata Sprengel extract and its major compound ent-kaurenoic acid display activity against bacteria present in endodontic infections.

The search for new, effective and safe antimicrobial compounds from plant sources has continued to play an important role in the maintenance of human health since ancient times. Such compounds can be used to help to eradicate microorganisms from the root canal system, preventing/healing periapical diseases. Mikania glomerata (Spreng.), commonly known as "guaco," is a native climbing plant from Brazil that displays a wide range of pharmacological properties. Many of its activities have been attributed to its phytochemical composition, which is mainly composed of diterpenes, such as ent-kaurenoic acid (KA). The present study evaluated the potential activity of an ent-kaurenoic-rich (KA) extract from Mikania glomerata (i.e. Mikania glomerata extract/MGE) and its major compound KA against bacteria that can cause endodontic infections. Time-kill assays were conducted and the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), anti-biofilm activity, and synergistic antimicrobial activity of MGE and KA were determined. The MGE exhibited MIC and MBC values, which ranged from 6.25 to 100 µg/mL and 12.5 to 200 µg/mL respectively. The MIC and MBC results obtained for the KA, ranged from 3.12 to 100 µg/mL and 3.12 to 200 µg/mL respectively. Time-kill and anti-biofilm activity assays conducted for KA at concentrations between 3.12 and 12.5 µg/mL exhibited bactericidal activity between 6 and 72 h of incubation and 50% inhibition of biofilm formation for Porphyromonas gingivalis (clinical isolate), Propionibacterium acnes (ATCC 6919), Prevotella nigrescens (ATCC 33563), P. melaninogenica (ATCC 25845), Aggregatibacter actinomycetemcomitans (ATCC 43717). For synergistic antimicrobial activity, KA combined with chlorhexidine dichlorohydrate (CHD) had an additive effect with increased efficacy against P. gingivalis (clinical isolate) compared to CHD alone. It was concluded that M. glomerata extract and its major compound ent-kaurenoic acid (KA) showed in vitro antibacterial activity, the latter being a potential biofilm inhibitory agent. They may play important roles in the search for novel sources of agents that can act against bacteria present in endodontic infections.

The emergence of the hyperinvasive vine, Mikania micrantha (Asteraceae), via admixture and founder events inferred from population transcriptomics.

Biological invasions that involve well-documented rapid adaptations to new environments provide unequalled opportunities for testing evolutionary hypotheses. Mikania micrantha Kunth (Asteraceae), a perennial herbaceous vine native to tropical Central and South America, successfully invaded tropical Asia in the early 20th century. It is regarded as one of the most aggressive weeds in the world. To elucidate the molecular and evolutionary processes underlying this invasion, we extensively sampled this weed throughout its invaded range in South-East and South Asia and surveyed its genetic structure using variants detected from population transcriptomics. Clustering results suggest that more than one source population contributed to this invasion. Computer simulations using genomewide genetic variation support a scenario of admixture and founder events during invasion. The genes differentially expressed between native and invasive populations were found to be involved in oxidative and high light intensity stress responses, pointing to a possible ecological mechanism of adaptation. Our results provide a foundation for further detailed mechanistic and population studies of this ecologically and economically important invasion. This line of research promises to provide new mitigation strategies for invasive species as well as insights into mechanisms of adaptation.

Effects of Mikania glomerata leaf extract on experimental Bothropoides jararaca envenomation in Wistar rats

Background: Bothropic envenomation represents the most common ophidic accident worldwide, compared to other snakebites of medical interest. Bothropic venom has proteolytic, vasculotoxic, clotting and/or hemorrhagic actions in animals and humans. Mikania glomerata is a plant found in the Brazilian Atlantic Forest with interesting medical properties that may be useful in ameliorating the effects of ophidic venom, and thus, improving response and outcome. Although Mikania is known to act through inhibition of cytolysins in the venom, there is a lack of consistent research data. The aim of this study was to evaluate the effect of M. glomerata in bothropic envenomation treatment. Materials, Methods & Results: Clinical, hematological, biochemical, and histopathological evaluations were performed following Bothropoides jararaca experimental envenomation in three groups of 18 Wistar rats each. Group VS was inoculated in the pelvic limb via intramuscular injection of bothropic venom and received specific anti-venom serum via intraperitoneal injection. Group VSM was similarly inoculated; it received anti-venom serum and a 10% aqueous extract of the Mikania glomerata plant orally. Group C was the control group and received saline solution alone. Evaluations were performed at 0.5 h (M1), 6 h (M2), and 24 h (M3) after venom inoculation. Animals from both inoculated groups (VS and VSM) showed significant clinical alterations (P < 0.05) manifested as discomfort, uneasiness, pain, and severe edema compared to control animals. Animals from inoculated groups also exhibited statistically significant leukocytosis with neutrophilia, and elevation of blood urea nitrogen and creatine kinase until 6 h after inoculation (P < 0.05 compared to control animals). An acute drop in body temperature was observed 6 h after inoculation (P < 0.05). [...]