Antitumoral Activity of Cecropia Pachystachya Leaves Extract in Vitro and in Vivo Model of Rat Glioma: Brain and Blood Effects.

The aim of this study was to investigate the antiglioma effect of Cecropia pachystachya Trécul (CEC) leaves extract against C6 and U87 glioblastoma (GB) cells and in a rat preclinical GB model. The CEC extract reduced in vitro cell viability and biomass. In vivo, the extract decreased the tumor volume approximately 62%, without inducing systemic toxicity. The deficit in locomotion and memory and an anxiolytic-like behaviors induced in the GB model were minimized by CEC. The extract decreased the levels of reactive oxygen species, nitrites and thiobarbituric acid reactive substances and increased the activity of antioxidant enzymes in platelets, sera and brains of GB animals. The activity of NTPDases, 5'-nucleotidase and adenosine deaminase (ADA) was evaluated in lymphocytes, platelets and serum. In platelets, ATP and AMP hydrolysis was reduced and hydrolysis of ADP and the activity of ADA were increased in the control, while in CEC-treated animals no alteration in the hydrolysis of ADP was detected. In serum, the reduction in ATP hydrolysis was reversed by CEC. In lymphocytes, the increase in the hydrolysis of ATP, ADP and in the activity of ADA observed in GB model was altered by CEC administration. The observed increase in IL-6 and decrease in IL-10 levels in the serum of GB animals was reversed by CEC. These results demonstrate that CEC extract is a potential complementary treatment to GB, decreasing the tumor size, while modulating aspects of redox and purinergic systems.

In Silico Studies of Four Compounds of Cecropia obtusifolia against Malaria Parasite.

Malaria is a disease that affects many people in the world. In Mexico, malaria remains an active disease in certain regions, particularly in the states of Chiapas and Chihuahua. While antimalarial effects have been attributed to some species of Cecropia in various countries, no such studies have been conducted in Mexico. Therefore, the objective of this study was to evaluate the in silico antimalarial activity of some active compounds identified according to the literature in the species of Cecropia obtusifolia, belonging to the Cecropiaceae family, such as ursolic acid, α-amyrin, chrysin, and isoorientin. These compounds were evaluated with specific molecular docking and molecular dynamics (MD) studies using three different malarial targets with the PDB codes 1CET, 2BL9, and 4ZL4 as well as the prediction of their pharmacokinetic (Pk) properties. Docking analysis revealed the following best binding energies (kcal/mol): isoorientin-1CET (-9.1), isoorientin-2BL9 (-8.8), and chrysin-4ZL4 (-9.6). MD simulation validated the stability of the complexes. Pharmacokinetics analysis suggested that the compounds would generally perform well if administered. Therefore, these results suggest that these compounds may be used as potential drugs for the treatment of malaria.

Intra- and Interspecies Differences of Two Cecropia Species from Tabasco, Mexico, Determined through the Metabolic Analysis and 1H-NMR-Based Fingerprinting of Hydroalcoholic Extracts.

The genus Cecropia is used in the traditional medicine of Tabasco, Mexico, in diabetes and hypertension treatments, mainly without distinction of the species. This contribution aimed to carry out the metabolic analysis and Proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy-based fingerprinting of the hydroalcoholic leaf extracts of Cecropia peltata (Cp) and Cecropia obtusifolia (Co) collected in five sub-regions of the State of Tabasco (Cp1, "Centro"; Cp2, "Chontalpa"; Cp3, "Pantanos"; Cp4, "Ríos" and Co5, "Sierra"). Firstly, the extracts were evaluated for their Total Phenol Content (TPC) and Total Flavonoid Content (TFC) by spectrophotometric methods. In addition, metabolic analysis was performed using High-Performance Liquid Chromatography with Diode-Array Detection HPLC-DAD, which allowed the quantification of the chemical markers: chlorogenic acid, isoorientin, and orientin, as well as a vitexin analog. Finally, metabolomic analysis was carried out based on the 1H-NMR spectra. The Cp4 extract (C. peltata from the "Ríos" sub-region) presented the highest values of TPC (155 ± 9.1 mg GAE/g E) and TFC (724 ± 22.2 mg RE/g E). The metabolic analysis was similar among the five samples; the highest concentrations of the four chemical markers were found in Cp3 (C. peltata from the "Pantanos" sub-region) for chlorogenic acid (39.8 ± 2.3 mg/g) and isoorientin (51.5 ± 2.9 mg/g), in Cp4 for orientin (49.9 ± 0.6 mg/g), and in Cp2 (C. peltata from the "Chontalpa" sub-region) for the vitexin analog (6.2 ± 0.2 mg/g). The metabolic analysis and the 1H-NMR fingerprint analysis showed intraspecies differences among the C. peltata samples and interspecies between C. peltata and C. obtusifolia, which were attributed to variations in the metabolite groups as well as in the proportion of sugars such as glucose and xylose.

Cecropia pachystachya Trécul: identification, isolation of secondary metabolites, in silico study of toxicological evaluation and interaction with the enzymes 5-LOX and α-1-antitrypsin.

Cecropia pachystachya Tréc., popularly known as embaúba, belongs to the Cecropiaceae family and is used by the native population in the treatment of bronchitis, asthma, high blood pressure, fever, and as a diuretic. The pharmacological actions including anti-inflammatory, antioxidant, cardiotonic and sedative were previously reported. The objective of this study was to (1) isolate and identify bioactive compounds extracted from the ethanolic extract of C. pachystachya roots (ERCP), as well as (2) verify the affinity of these metabolites with the enzymes 5-lipoxygenase (5-LOX) and α-1-antitrypsin through in silico tests. Isolation and/or identification were performed using GC-MS, HPLC, Infrared (IR), and nuclear magnetic resonance (NMR) techniques. After isolation and identification of the active compounds, these substances were subjected to the in silico investigation that proceeded by performing PreADMET simulations and molecular docking calculations. The bioactive compounds identified were 1-(+)-ascorbic acid 2,6-dihexadecanoate, ethyl hexadecanoate, ethyl (9E,12E)-octadec-9,12-dienoate, ethyl (Z)-octadec-9-enoate and ethyl octadecanoate by GC-MS; chlorogenic acid, catechin, epicatechin, syringaldehyde by HPLC; ß-sitosterol, sitostenone, beccaridiol, tormentic acid, lupeol, α- and ß-amyrin by classical chromatography, IR, 1H and 13C NMR techniques. The ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties were determined for each bioactive compound. Tormentic acid demonstrated a greater affinity for 5-LOX enzyme while sitostenone demonstrated a higher affinity for the α-1-antitrypsin enzyme. Our findings demonstrated a diverse range of secondary metabolites isolated from C. pachystachya that showed relevant interactions with the enzymes 5-LOX and α-1-antitrypsin. Thus, "embaúba" may be employed in in vivo experimental studies as a form of alternative treatment for chronic lung diseases.Abbreviations: ADT: Autodock Tools; BBB: Blood-brain barrier; CaCo2: Human colonic adenocarcinoma cells; CC: Classic/open Column; TLC: Thin Layer Chromatography; CD40: Differentiation Cluster 40; CENAUREMN: Centro Nordestino de Aplicação e Uso da Ressonância Magnética Nuclear; GC-MS: Gas Chromatography coupled to mass spectrometry; HPLC: High-Perfomance Liquid Chromatography; CYP2C9, CYP2C19, CYP2D6 and CYP3A4: Cytochrome P450 isoenzymes; COPD: Chronic Obstructive Pulmonary Disease; DRX-500: X-Ray Diffraction - 500; ERCP: Ethanolic extract of the roots of C. pachystachya; FAPEPI: Fundação de Amparo à Pesquisa do Piauí; HIA: Human Intestinal Absorption; IR: Infrared; Ki: Inhibition constant; 5-LOX: 5-Lipoxygenase; mM: miliMolar; nM: nanoMolar; OECD423: acute toxic class method; PDB: Protein Data Bank; P-gP: P-glycoprotein; PM2,5: Small inhalable particles 2,5; PPB: Plasm Protein Binding; PreADMET: Prediction Absorption, Distribution, Metabolization, Excretion and Toxicity; NMR: Nuclear Magnetic Resonance; +S9: with metabolic activation; -S9: no metabolic activation; SisGen: Sistema Nacional de Gestão de Patrimônio Genético e do Conhecimento Tradicional Associado; RT: Retention time; TA100: Ames test with TA100 cells line; TA1535: Ames test with cells of the TA1535 cell line; UESPI: State University of Piauí; V79: lung fibroblast cells; ΔG: Gibbs free energy (Kcal/mol); µM: microMolar.

Cecropia pachystachya Trécul: a promising ingredient for skin-whitening cosmetics

Abstract Because it promotes the lightening of pigment spots, tyrosinase inhibition is one of the mechanisms of depigmenting cosmetic products. Considering the adverse effects produced by synthetic depigmenting actives, the search for new therapeutic options is desirable, and plant extracts are possible candidates for hyperpigmentation treatment. Glycolic extracts of Cecropia pachystachya Trécul are, therefore, the focus of this study. Its chemical characterization, antioxidant activity, tyrosinase inhibition, and cell viability were evaluated. Glycolic extracts were obtained by macerating the leaves of C. pachystachya in grain alcohol and glycerin or propylene glycol. Both had a similar chemical constitution, the glycerin being more efficient in concentrating phenolic compounds and flavonoids. Analyses by UHPLC-MS detected quinic acid, chlorogenic acid isomers, proanthocyanidin dimers type B and C, catechin/epicatechin, orientin/isoorientin, isoorientin 2"-O-xyloside, vitexin/isovitexin, and rutin. 5-O-caffeoylquinic acid was then quantified was then quantified, with predominance in the extract produced with propylene glycol. These extracts showed a high antioxidant capacity by the method of DPPH, ß-carotene, and nitric oxide. As for depigmenting activity, both extracts were able to inhibit tyrosinase. Cell viability assay also revealed that the extracts could safely be used in concentrations of ≤ 125 µg/mL. Thus, this study demonstrated for the first time that the glycolic extracts of C. pachystachya have promising chemical and biological characteristics for the development of a multifunctional cosmetic with antioxidant and tyrosinase-inhibition activities

Pentacyclic Triterpene Profile and Its Biosynthetic Pathway in Cecropia telenitida as a Prospective Dietary Supplement.

Promising research over the past decades has shown that some types of pentacyclic triterpenes (PTs) are associated with the prevention of type 2 diabetes (T2D), especially those found in foods. The most abundant edible sources of PTs are those belonging to the ursane and oleanane scaffold. The principal finding is that Cecropia telenitida contains abundant oleanane and ursane PT types with similar oxygenation patterns to those found in food matrices. We studied the compositional profile of a rich PT fraction (DE16-R) and carried out a viability test over different cell lines. The biosynthetic pathway connected to the isolated PTs in C. telenitida offers a specific medicinal benefit related to the modulation of T2D. This current study suggests that this plant can assemble isobaric, positional isomers or epimeric PT. Ursane or oleanane scaffolds with the same oxygenation pattern are always shared by the PTs in C. telenitida, as demonstrated by its biosynthetic pathway. Local communities have long used this plant in traditional medicine, and humans have consumed ursane and oleanane PTs in fruits since ancient times, two key points we believe useful in considering the medicinal benefits of C. telenitida and explaining how a group of molecules sharing a closely related scaffold can express effectiveness.

Effects of two dietary fiber levels on nutrient digestibility and intestinal fermentation products in captive brown howler monkeys (Alouatta guariba).

Herbivorous primates present a selective consumption profile and morphological adaptations to use the fibrous fraction of their diets. Brown howler monkeys (Alouatta guariba) are generalist herbivores; however, when kept under human care, they usually receive diets rich in fruits and with insufficient amounts of fiber. Thus, the objective of this study was to evaluate the effects of two levels of neutral detergent fiber (NDF) in howlers on apparent total tract digestibility coefficients (ATTDC), fecal consistency, and intestinal fermentation products. A group of 26 adult howler monkeys, 13 males and 13 females, were fed two diets formulated to have 33% or 40% NDF for 11 days, according to a randomized block design (N = 26). The block factor was the enclosures with one, two, or three individuals (each enclosure corresponded to an experimental unit), totaling in eight replicates per treatment. There were no differences in dry matter and nutrients intake between treatments (p > 0.05). The diet with 33% NDF resulted in higher (p < 0.05) ATTDC of crude protein and crude energy. However, lower fecal concentrations of short-chain fatty acids (SCFA) and dry matter were observed in the treatment with 33% NDF in contrast to the 40% NDF group (p < 0.05). We recommend the inclusion of higher fiber levels (40% NDF) in the diet of howler monkeys since there is evidence of greater production of SCFA and improvement in fecal consistency.

Insights Into the Ecological Role of Pseudomonas spp. in an Ant-plant Symbiosis.

In the myrmecophytic mutualistic relationship between Azteca ants and Cecropia plants both species receive protection and exchange nutrients. The presence of microorganisms in this symbiotic system has been reported, and the symbiotic role of some fungi involved in the myrmecophytic interactions has been described. In this work we focus on bacteria within this mutualism, conducting isolations and screening for antimicrobial activities, genome sequencing, and biochemical characterization. We show that Pantoea, Rhizobium, Methylobacterium, Streptomyces and Pseudomonas are the most common cultivable genera of bacteria. Interestingly, Pseudomonas spp. isolates showed potent activity against 83% of the pathogens tested in our antimicrobial activity assays, including a phytopathogenic fungus isolated from Cecropia samples. Given the predicted nitrogen limitations associated with the fungal patches within this myrmecophyte, we performed nitrogen fixation analyses on the bacterial isolates within the Proteobacteria and show the potential for nitrogen fixation in Pseudomonas strains. The genome of one Pseudomonas strain was sequenced and analyzed. The gene cluster involved in the biosynthesis of cyclic lipodepsipeptides (CLPs) was identified, and we found mutations that may be related to the loss of function in the dual epimerization/condensation domains. The compound was isolated, and its structure was determined, corresponding to the antifungal viscosinamide. Our findings of diazotrophy and production of viscosinamide in multiple Pseudomonas isolates suggests that this bacterial genus may play an important role in the Cecropia-Azteca symbiosis.

Cecropia hololeuca: A new source of compounds with potential anti-inflammatory action.

Our objective is to investigate the phytochemical components, antioxidant capacity and in vitro and in vivo anti-inflammatory action from Cecropia hololeuca bark aqueous extract (AECh). The chemical characterization of AECh was performed through CE-UV, FTIR and NMR Spectroscopy. In vitro assays were performed with the AECh on murine macrophages J774A.1 cells in order to analyse cell viability, NO, TNF-α and IL-1ß productions and the in vivo anti-inflammatory potential in acute carrageenan paw oedema in mice. The AECh showed a decrease in the production of NO, TNF-α and IL-1ß, without altering the cell viability and reduction of the paw thickness in the 2nd, 3rd and 4th hour. The extract presented 72% free radical scavenging, 0.60% flavonoid content and showed the presence of gallic acid, caffeic acid and catechin as major constituents. The C. hololeuca bark extract showed important antioxidant and anti-inflammatory activity, emphasizing the industrial and pharmacological potential of this plant.

Integrated Analysis of the Transcriptome and Metabolome of Cecropia obtusifolia: A Plant with High Chlorogenic Acid Content Traditionally Used to Treat Diabetes Mellitus.

This investigation cultured Cecropia obtusifolia cells in suspension to evaluate the effect of nitrate deficiency on the growth and production of chlorogenic acid (CGA), a secondary metabolite with hypoglycemic and hypolipidemic activity that acts directly on type 2 diabetes mellitus. Using cell cultures in suspension, a kinetics time course was established with six time points and four total nitrate concentrations. The metabolites of interest were quantified by high-performance liquid chromatography (HPLC), and the metabolome was analyzed using directed and nondirected approaches. Finally, using RNA-seq methodology, the first transcript collection for C. obtusifolia was generated. HPLC analysis detected CGA at all sampling points, while metabolomic analysis confirmed the identity of CGA and of precursors involved in its biosynthesis. Transcriptome analysis identified differentially expressed genes and enzymes involved in the biosynthetic pathway of CGA. C. obtusifolia probably expresses a key enzyme with bifunctional activity, the hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase and hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HQT/HCT), which recognizes shikimic acid or quinic acid as a substrate and incorporates either into one of the two routes responsible for CGA biosynthesis.

Cytotoxic effects of a triterpene-enriched fraction of Cecropia pachystachya on the human hormone-refractory prostate cancer PC3 cell line.

BACKGROUND: Prostate cancer (PCa) is the most diagnosed invasive cancer and a leading cause of death in men in western countries. Most patients initially respond to androgen deprivation but finally develop hormone-refractory disease, which results in advanced clinical failure and death. Since hormone-refractory disease is highly radiotherapy and chemotherapy resistant, increasing interest has been placed on finding novel therapies for this advanced type of Pca. PURPOSE: The potential cytotoxic effects of the crude extract and fractions obtained from the leaves of Cecropia pachystachya Trécul on different human cancer cell lines were investigated. Additionally, the mechanism of cell death induction of the most active sample (triterpene-enriched fraction, TEF) on the human hormone-refractory prostate PC3 cell line was examined. METHODS: Sulforhodamine B assay was used to measure the viability of human tumor and non-tumor cell lines. To elucidate the mechanism of PC3 cells death induced by TEF, different methodological approaches were used: cell cycle analysis and annexin V/PI staining, nuclear morphological analysis, and senescence-associated-ß-galactosidase assay. Moreover, the mitochondrial membrane potential was measured, and the long-term effects of TEF on PC3 cells were evaluated. RESULTS: TEF exerted cytotoxic effects on PC3 cells but not on human non-tumor cells. The analysis of nuclear morphology of PC3 cells treated with TEF increased the number of cells with large and regular nuclei suggesting senescence induction, which was supported by ß-galactosidase overexpression. Regarding PC3 cells cycle, TEF reduced the number of cells in G1 phase and increased that in sub G0/G1. Apoptosis was not involved in PC3 cell death. However, there was a decrease in mitochondrial membrane potential without the participation of reactive oxygen species (ROS) in the cytotoxic effects detected. Furthermore, there was a decrease in the number of viable cells able to duplicate after long-term TEF treatment. CONCLUSIONS: The results showed the in vitro cytotoxic potential of the triterpene-enriched fraction obtained from the leaves of C. pachystachya on human prostate cancer PC3 cell line.

Serjanic Acid Improves Immunometabolic Markers in a Diet-Induced Obesity Mouse Model.

Plant extracts from Cecropia genus have been used by Latin-American traditional medicine to treat metabolic disorders and diabetes. Previous reports have shown that roots of Cecropia telenitida that contains serjanic acid as one of the most prominent and representative pentacyclic triterpenes. The study aimed to isolate serjanic acid and evaluate its effect in a prediabetic murine model by oral administration. A semi-pilot scale extraction was established and serjanic acid purification was followed using direct MALDI-TOF analysis. A diet induced obesity mouse model was used to determine the impact of serjanic acid over selected immunometabolic markers. Mice treated with serjanic acid showed decreased levels of cholesterol and triacylglycerols, increased blood insulin levels, decreased fasting blood glucose and improved glucose tolerance, and insulin sensitivity. At transcriptional level, the reduction of inflammation markers related to adipocyte differentiation is reported.

Mechanism of the Hypoglycemic Activity and Hepatoprotective Effect of the Aqueous Extract of Cecropia obtusifolia Bertol.

In Central and South American traditional medicine, people use Cecropia obtusifolia Bertol (Cecropiaceae) for the treatment of diabetes mellitus. However, its hypoglycemic action mechanism at pancreatic and liver level has been poorly explored. The present research aimed to establish the influence of the aqueous extract of C. obtusifolia, standardized in its content of chlorogenic acid, on insulin secretion in RINm5F cells and over the liver carbohydrates and lipids metabolism, and to determine concomitantly its hepatoprotective effect on mice with streptozotocin-induced diabetes. In RINm5F cells, concentrations 5, 50, 100, and 200 µg/mL of aqueous extract of C. obtusifolia were used to determine [Ca2+]i and insulin secretion. In an acute study, the extract was administered at doses of 500 mg/kg. In another test (subacute), the extract was daily administrated to diabetic mice (200 mg/kg/day) for 30 days. Blood glucose levels and other biochemical parameters were determined, and a liver histological analysis was performed. In RINm5F cells, C. obtusifolia increased [Ca2+]i and insulin secretion, whereas in diabetic mice exhibited acute and subacute hypoglycemic effects. Daily administration of C. obtusifolia to diabetic mice also increased liver glycogen storage and glycogen synthase levels, without apparent changes in gluconeogenesis. Besides, it increased peroxisome proliferator-activated receptor-α (PPAR-α) and long-chain-fatty-acid-CoA ligase 1 (ACSL-1) expression and reduced triglycerides, transaminases (alanine aminotransferase and aspartate aminotransferase), and collagen fibers, modifying anti-inflammatory (adiponectin and interleukin-10) and inflammatory (tumor necrosis factor-α) cytokines in serum. Therefore, the hypoglycemic effect of C. obtusifolia implicates a dual action, promoting insulin secretion, liver glycogen accumulation, and hepatoprotection by decreasing collagen fibers and inflammatory markers, whereas it improves lipid metabolism, due in part to PPAR-α.

Hidden gems in museum cabinets: new species and new distributional records of Scolytodes (Coleoptera: Scolytinae).

Twenty-one new species of Scolytodes Ferrari are described from Central and South America: Scolytodes cavus Jordal, sp. nov., Scolytodes excavatus Jordal, sp. nov., Scolytodes speculofrons Jordal, sp. nov., Scolytodes potens Jordal, sp. nov., Scolytodes minimus Jordal, sp. nov., Scolytodes setosicauda Jordal, sp. nov., Scolytodes monticola Jordal, sp. nov., Scolytodes aureifrons Jordal, sp. nov., Scolytodes piliscapus Jordal, sp. nov., Scolytodes curvicostatus Jordal, sp. nov., Scolytodes vellus Jordal, sp. nov., Scolytodes granulatus Jordal, sp. nov., Scolytodes bipilosus Jordal, sp. nov., Scolytodes asperatus Jordal, sp. nov., Scolytodes flavifrons Jordal, sp. nov., Scolytodes capillus Jordal, sp. nov., Scolytodes rufus Jordal, sp. nov., Scolytodes johnsoni Jordal, sp. nov., Scolytodes longipilus Jordal, sp. nov., Scolytodes prolatus Jordal, sp. nov., and Scolytodes bicarinatus Jordal, sp. nov. Females of two species were associated with males and described for the first time: Scolytodes pelicipennis (Schedl, 1952) from Mexico, and Scolytodes venustulus Wood, 1967 from Costa Rica. New synonymies were inferred in four species: Scolytodes bolivianus Eggers, 1928 (=Scolytodes aequipunctatus Eggers, 1943 syn. nov.); Scolytodes major Eggers, 1928 (=Scolytodes argentinensis Eggers, 1943 syn. nov.; =Scolytodes boliviensis Eggers, 1928; =Scolytodes similis Eggers, 1928 syn. nov.); Scolytodes ingae (Blackman, 1943), stat. nov. (=Scolytodes trigonus Jordal, 2013 syn. nov.); Scolytodes canalis Wood, 1974 (=Scolytodes amabilis Wood, 1975 syn. nov.). New country records are reported for 13 species: S. alni Wood, 1969 (Panama), S. bolivianus (Brazil), S. canalis Wood, 1974 (Guatemala), S. chapuisi Wood, 1977 (Peru, Brazil), S. costabilis Wood (Costa Rica), S. elongatus Schedl, 1935 (Panama), S. immanis Wood, 1969 (Panama), S. ingae (Blackman, 1943) (Panama), S. irazuensis Wood, 1969 (Panama), S. major (Honduras, Brazil), S. pelicipennis (Schedl, 1952) (Mexico), S. triangulus Jordal, 1998 (Panama), S. unipunctatus (Blandford, 1897) (Panama, Venezuela), S. venustulus Wood, 1967 (Costa Rica).

Vasorelaxant effect of standardized extract of Cecropia glaziovii Snethl encapsulated in PLGA microparticles: In vitro activity, formulation development and release studies.

In this study, the standardized dry extract of C. glaziovii (SDE) provided a significant vasorelaxant effect after contractions induced by phenylephrine in rat aortic rings in an endothelium-dependent manner, confirming that endothelial factors are needed to stimulate this response. A vasorelaxation close to that of acetylcholine was achieved, justifying the development of new formulations for this plant material. In this context, microparticles were selected to encapsulate SDE and the double emulsion technique was considered because of the hydrophilic nature of plant material. Two experimental designs were applied. Firstly, the effect of formulation parameters on particle size, size distribution and encapsulation efficiency (EE) was evaluated. As low EE was achieved, the effect of the osmotic pressure of the external phase was evaluated in a second experimental design. The presence of the osmotic agent (NaCl) impacted positively on the EE and slower in vitro release profile was obtained, which is desired in controlled release systems. The formation of denser and less porous particle surface, which was identified by SEM analysis, contributes to explain these findings. Microstructures showed to be a promising delivery system for the C. glaziovii SDE considering that a sustained release was achieved.

Detection, ultrastructure and phylogeny of Sclerorhabditis neotropicalis (Nematoda: Rhabditidae) nematodes associated with the Azteca ant-Cecropia tree symbiosis / Detección, ultraestructura y filogenia de los nemátodos Sclerorhabditis neotropicalis (Nematoda: Rhabditidae) asociados con la simbiosis de la hormiga Azteca y el árbol

Abstract The nematodes involved in ant-plant symbioses have been poorly studied, creating an area of research that is now receiving increasing attention. One example is Sclerorhabditis neotropicalis that inhabit the nests of Azteca ants in Cecropia trees. The goal of the present study is to increase our knowledge about these organisms and their possible symbiotic relationship with the ants and their host plant. The samples were collected during 2011-2013 in different locations in Costa Rica: Parque Nacional Carara (Puntarenas province), San Pedro de Montes de Oca and Highway 27 (San Jose) and La Selva Biological Station (Sarapiquí, Heredia). We examined 576 internodes from 23 plants, comprising four species of Cecropia (including one non-myrmecophyte) and five Azteca species. S. neotropicalis was found in all but one of the Azteca/Cecropia combinations examined. Queen and worker ants were placed in water in order to dislodge nematodes and the results were largely positive, suggesting that alate queens carry S. neotropicalis between trees and that workers carry them between internodes. In addition to transport, preliminary observations suggest that the ants provide masses of scraped parenchyma (possibly fertilized with feces) as a substrate for bacteria upon which the nematodes presumably feed. We also found S. neotropicalis associated with Azteca in another myrmecophyte, Cordia alliodora, suggesting that this ant/nematode association is not restricted to Cecropia. SEM photographs of S. neotropicalis were provided to supplement the original description of this species, and molecular phylogenetic analyses employing three different markers suggest that Sclerorhabditis forms a well-supported clade of bacteria-feeding nematodes associated with Azteca. Indirect evidence suggests that this is a mutualistic association, but the benefits to the ants remain unknown. Rev. Biol. Trop. 66(1): 368-380. Epub 2018 March 01.

Resumen Cecropia. Los nemátodos asociados a la simbiosis entre plantas y hormigas han sido poco estudiados, creando un área de investigación que está recibiendo cada vez más atención. Un ejemplo de esto es Sclerorhabditis neotropicalis, que habita en los nidos de las hormigas Azteca asociadas a árboles de Cecropia. El objetivo de este trabajo es incrementar el conocimiento sobre esta especie de nemátodo y su relación con Azteca-Cecropia. Las muestras fueron recolectadas durante 2011-2013 en diferentes localidades de Costa Rica: Parque Nacional Carara (Puntarenas), San Pedro de Montes de Oca y Ruta 27 (San José) y la Estación Biológica La Selva (Sarapiquí, Heredia). Se examinaron 576 entrenudos de 23 plantas, comprendiendo cuatro especies de Cecropia (incluyendo una especie no mirmecófita) y cinco especies de Azteca. Se encontraron individuos de S. neotropicalis en todas las combinaciones de Azteca/Cecropia examinadas. Las hormigas reinas y trabajadoras fueron colocadas en agua para observar los nematodos y los resultados fueron en gran medida positivos, lo cual sugiere que las reinas aladas transportan S. neotropicalis hacia nuevos árboles y que las trabajadoras los transportan entre los entrenudos. Además del transporte, las observaciones preliminares insinúan que las hormigas proporcionan una masa de parénquima raspado (posiblemente fertilizado con heces) como sustrato para bacterias que luego servirán de alimento a los nematodos. Adicionalmente, encontramos S. neotropicalis asociado con Azteca en otro tipo de árbol, Cordia alliodora, sugiriendo que esta asociación de hormigas/nematodos no se limita a Cecropia. Las fotografías de microscopía electrónica de S. neotropicalis obtenidas en este trabajo complementan la descripción original de esta especie. Por otra parte, los análisis filogenéticos moleculares sugieren que Sclerorhabditis forma un clado bien definido de nematodos bacteriófagos asociados con Azteca. La evidencia indirecta apunta a que se trata de una asociación mutualista, pero los beneficios para las hormigas siguen siendo desconocidos.

LC/QTOF profile and preliminary stability studies of an enriched flavonoid fraction of Cecropia pachystachya Trécul leaves with potential antidepressant-like activity.

There is increasing interest in natural antioxidants that are candidates for the prevention of brain damage occurring in major depressive disorders. Cecropia pachystachya is a tropical tree species of Central and South America and a rich source of polyphenols, particularly flavonoids. The aim of this study was to characterize the flavonoid profile of an enriched flavonoid fraction of C. pachystachya (EFF-Cp) and evaluate the antidepressant-like effects of its acute administration in behavior, cytokine levels, oxidative stress and energy metabolism parameters. The EFF-Cp chemical characterization was performed by HPLC/DAD and LC/QTOF. The antidepressant-like effects were performed by the forced swimming test, splash test and open field test. EFF-Cp revealed 15 flavonoids, including seven new glycosyl flavonoids for C. pachystachya. Quantitatively, EFF-Cp showed isoorientin (43.46 mg/g), orientin (23.42 mg/g) and isovitexin (17.45 mg/g) as major C-glycosyl flavonoids. In addition, EFF-Cp at doses 50 and 100 mg/kg reduced the immobility time in the forced swimming test, without changing the locomotor activity and grooming time. In addition, EFF-Cp was able to prevent the oxidative damage in some brain areas. In conclusion, the results of this study suggest that EFF-Cp exerts antidepressant-like effects with its antioxidant properties.

Recurrent breakdowns of mutualisms with ants in the neotropical ant-plant genus Cecropia (Urticaceae).

Mutualisms could be evolutionarily unstable, with changes in partner abundances or in the spatial context of interactions potentially promoting their dissolution. We test this prediction using the defense mutualisms between species of the Neotropical genus Cecropia and Azteca ants. A new, multigene phylogeny with representatives of all five genera of Cecropieae (most of them from the Neotropics) and half of the 61 species of Cecropia shows the West African endemic Musanga (2spp.) as sister to Cecropia, implying dispersal from the Neotropics to Africa, with a molecular clock suggesting that this occurred about 23Mya. Cecropia, a genus of neotropical pioneer trees, started diversifying ca. 8Mya. We infer a single origin of specialized symbiosis with Azteca within Cecropia, eight complete losses of this symbiosis, and a potential partner shift involving the replacement of Azteca by Neoponera luteola ants. Niche space modeling based on geo-referenced occurrences of over 9000 collections representing 58 of the 61 species of Cecropia, together with several comparative analyses, implies that mutualism loss is concentrated at high altitudes and on Caribbean islands, with the surprisingly frequent breakdowns potentially facilitated by low species-specificity of interacting Cecropia and Azteca mutualists.

Genotoxic, mutagenic and antigenotoxic effects of Cecropia pachystachya Trécul aqueous extract using in vivo and in vitro assays.

ETHNOPHARMACOLOGICAL RELEVANCE: Cecropia pachystachya is a medicinal plant native to South and Central Americas used to treat asthma and diabetes. AIM OF THE STUDY: In this study, we evaluated the genotoxic, mutagenic and antigenotoxic effects of crude aqueous extract of C. pachystachya (CAE-Cp) leaves. MATERIAL AND METHODS: CAE-Cp was analyzed by the Folin-Ciocalteu method to determine total phenolic and tannin contents. High performance liquid chromatography (HPLC) was used to identify major compounds. Distinct tissues from female and male adult mice were treated with 500-2000mg/kg of CAE-Cp by gavage for the comet assay and micronucleus test analyses. In addition, peripheral blood slides of the group treated with 2000mg/kg CAE-Cp were analyzed 3, 6, and 24h after treatment and were exposed to hydrogen peroxide (ex vivo) to evaluate the genotoxic effect using the comet assay. The Salmonella/microsome assay was carried out against to TA100, TA98, TA97a, TA102, and TA1535 strains in presence and absence of the S9 mix. RESULTS: HPLC showed the presence of chlorogenic acid, isoorientin, orientin, and isovitexin as major compounds. Total phenolic and tannin contents were, respectively, 305.6±0.80 and 144.6±19.04mg of gallic acid equivalent/g of extract. Brain DNA damage was observed in all groups treated with CAE-Cp. The H2O2 challenge indicated genotoxic effect only 6h after the administration of the extract. No increase was detected in micronucleus frequency for any group treated with the extract. Mutagenic effects were detected by Salmonella/microsome assay only in TA102 strain without S9 mix at higher doses. CONCLUSION: The results obtained indicate that CAE-Cp was genotoxic to brain tissue. This result is supported by other papers, showing that compounds present in this extract can cross the blood-brain barrier and act on central nervous system.

Anti-inflammatory and antioxidant activities of aqueous extract of Cecropia glaziovii leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Cecropia glaziovii Sneth leaves extract is widely used as a traditional folk medicine in Brazil, especially for the treatment of diabetes, and as an antihypertensive and antiinflammatory agent. AIM OF THE STUDY: To investigate the anti-inflammatory and antioxidant properties of crude aqueous extract (CAE) of C. glaziovii leaves. MATERIALS AND METHODS: The in vivo anti-inflammatory and antioxidant effect of the CAE (10-300mg/kg, intragastrically) was investigated in the animal model of pleurisy. The cell migration, proinflammatory cytokines (TNF-α, IL-1ß and IL-6), nitrite/nitrate concentration, myeloperoxidase (MPO) activity, oxidative damage in lipids and proteins, lactate dehydrogenase (LDH) activity and total protein content were also analyzed. Furthermore, the in vitro antioxidant activity of CAE was evaluated by the inhibition of formation of thiobarbituric acid reactive substances (TBARS), induced by free radical generators (H2O2, FeSO4 and AAPH) on a lipid-rich substrate. Hence, the chemical characterizarion of CAE by HPLC was therefore performed. The results showed that the inflammatory process caused by the administration of carragenin (Cg) into the pleural cavity resulted in a substantial increase in inflammatory parameters and oxidative damage. These levels seems to be reversed after CAE treatment in animals with similar results to Dexamethasone (Dex) treatment. Further, the CAE was effective in reducing proinflammatory cytokines, cell infiltrate, MPO activity, nitrite/nitrate concentration, LDH activity, and total protein levels with concomitant attenuation of all parameters associated with oxidative damage induced by Cg. Finally, the CAE presented in vitro antioxidant activity induced by free radical generators at all the concentrations investigated. HPLC analysis confirmed the presence of chlorogenic acid and C-glycosylflavonoids (isoorientin and isovitexin) as the major compounds of the CAE. CONCLUSION: CAE of C. glaziovii exerts significant antiinflammatory and antioxidant activities and this effect can be attributed, at least in part, to the presence of chlorogenic acid and the C-glycosylflavonoids.