Hydrophilic Scaffolds Containing Extracts of Stryphnodendron adstringens and Abarema cochliacarpa for Wound Healing: In Vivo Proofs of Concept.

The present work aimed to evaluate the healing effect of hydrophilic polymeric resorbable biomembrane scaffolds containing plant extracts obtained from two different species, both popularly known as Stryphnodendron adstringens or Barbatimão. The hydrogel-based scaffolds were characterized and submitted to biological tests using Wistar rats to evaluate their healing capacity. The wound retraction index and the evaluation of the inflammatory process and tissue collagenization were recorded. The extracts showed antioxidant activity with IC50 between 10 and 20 µg/mL (DPPH assay) and 4-6 mmol Trolox/g (FRAP assay). The extract of Stryphnodendron adstringens (SA) presented gallocatechin, epigallocatechin, and O-methylpigalocatechin, while the extract of Abarema cochliacarpa (AC) presented catechin, dimers of procyanidins, di-O-hydroxide, O-deoxyhexosi-hexoside, and epicatechin. The membranes containing SA extract (GELSA) were more rigid, with a more intense color, but less thick, with a more compact structure and few pores. The membranes containing AC extract (GELAC) presented a mechanical profile like the gelatin membrane (GEL), with greater permeability to water vapor. The GELAC and GELSA membranes showed similar thermal degradation profiles. The wounds treated with the membranes containing the extracts obtained high levels of retraction of the wounds with values around 60% and 80% in three and seven days, respectively. These data indicate that the compounds of both species have promising biological activities in the repair process, showing that the extracts accelerated the healing process due to the lower intensity of the inflammatory reaction and the presence of compounds such as catechin and epigallocatechin.

Using Cell Cultures for the Investigation of Treatments for Attention Deficit Hyperactivity Disorder: A Systematic Review.

BACKGROUND: Advances in basic and molecular biology have promoted the use of cell cultures in a wide range of areas, including the evaluation of drug efficacy, safety and toxicity. OBJECTIVE: This article aims to provide a general overview of the methodological parameters of cell cultures used to investigate therapeutic options for Attention Deficit Hyperactivity Disorder. METHOD: A systematic search was performed in the electronic databases PubMed, Scopus, and DOAJ. In vitro experimental studies using cell cultures were included. RESULTS: A total of 328 studies were initially identified, with 16 included for qualitative synthesis. Seven studies used neuronal cells (SH-SY5Y neuroblastoma and PC12 cell line) and nine used nonneuronal cells. All the studies described the culture conditions, but most studies were inconsistent with regard to reporting results and raw data. Only one-third of the studies performed cell viability assays, while a further 30% conducted gene expression analysis. Other additional tests included electrophysiological evaluation and transporter activity. More than 50% of the studies evaluated the effects of drugs such as methylphenidate and atomoxetine, while plant extracts were assessed in four studies and polyunsaturated fatty acids in one. CONCLUSION: We suggested a flowchart to guide the planning and execution of studies, and a checklist to be completed by authors to allow the standardized reporting of results. This may guide the elaboration of laboratory protocols and further in vitro studies.

Morpho-anatomical characters of Limonium brasiliense leaves

Abstract Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, is a coastal herb of the Southern Brazilian states and Rio de Janeiro State. In folk medicine, it is used in the treatment of female genitourinary tract infections and menstrual cycle irregularities. The aim of this study was to describe morpho-anatomical characters with details on venation pattern of L. brasiliense leaves, collected on the coast of Rio Grande do Sul State, in Southern Brazil. Leaf samples fixed in FAA50 (5% formaldehyde, 5% acetic acid and 90% ethanol: water mixture 50:50 v/v) and 1% glutaraldehyde and preserved in 70% ethanol, were histologically sectioned and analyzed by light and scanning electron microscopy. The L. brasiliense leaves were simple, entire, incomplete, oboval-lanceolate, venation penninervous and showed rosulated phyllotaxy, entire and revolute margin, with apex rounded to slightly retuse, base attenuate and venation pattern brochidodromous. On microscopic analysis, these leaves showed a thick and striated cuticle, salt glands, isobilateral mesophyll thicker in the apical region of the leaf blade, abundant aerenchyma in the petiole and main vein, collateral vascular bundles surrounded by fiber-sclereids, lipid substances in the cuticle and polyphenols in phloem cells.

Proanthocyanidin Polymer-Rich Fraction of Stryphnodendron adstringens Promotes in Vitro and in Vivo Cancer Cell Death via Oxidative Stress.

Cervical cancer is the fourth most common cancer that affects women, mainly through human papilloma virus (HPV) infection with high-risk HPV16 and HPV18. The present study investigated the in vitro anticancer activity and mechanism of action of a proanthocyanidin polymer-rich fraction of Stryphnodendron adstringens (F2) in cervical cancer cell lines, including HeLa (HPV18-positive), SiHa (HPV16-positive), and C33A (HPV-negative) cells, and also evaluated in vivo anticancer activity. In vitro, cell viability was determined by the MTT assay. Cell migration was determined by the wound healing assay. The mechanism of action was investigated by performing ultrastructural analysis and evaluating reactive oxygen species (ROS) production, mitochondrial metabolism, lipoperoxidation, BCL-2 family expression, caspase expression, and DNA and cell membrane integrity. In vivo activity was evaluated using the murine Ehrlich solid tumor model. F2 time- and dose-dependently reduced cell viability and significantly inhibited the migration of cervical cancer cells. HeLa and SiHa cells treated with F2 (IC50) exhibited intense oxidative stress (i.e., increase in ROS and decrease in antioxidant species) and mitochondrial damage (i.e., mitochondrial membrane potential depolarization and a reduction of intracellular levels of adenosine triphosphate). Increases in the Bax/BCL-2 ratio and caspase 9 and caspase 3 expression, were observed, with DNA damage that was sufficient to trigger mitochondria-dependent apoptosis. Cell membrane disruption was observed in C33A cells (IC50 and IC90) and HeLa and SiHa cells (IC90), indicating progress to late apoptosis/necrosis. The inhibition of ROS production by N-acetylcysteine significantly suppressed oxidative stress in all three cell lines. In vivo, F2 significantly reduced tumor volume and weight of the Ehrlich solid tumor, and significantly increased lipoperoxidation, indicating that F2 also induces oxidative stress in the in vivo model. These findings indicate that the proanthocyanidin polymer-rich fraction of S. adstringens may be a potential chemotherapeutic candidate for cancer treatment.

The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain.

One of the most important postharvest plant pathogens that affect strawberries, grapes and tomatoes is Botrytis cinerea, known as gray mold. The fungus remains in latent form until spore germination conditions are good, making infection control difficult, causing great losses in the whole production chain. This study aimed to purify and identify phenazine-1-carboxylic acid (PCA) produced by the Pseudomonas aeruginosa LV strain and to determine its antifungal activity against B. cinerea. The compounds produced were extracted with dichloromethane and passed through a chromatographic process. The purity level of PCA was determined by reversed-phase high-performance liquid chromatography semi-preparative. The structure of PCA was confirmed by nuclear magnetic resonance and electrospray ionization mass spectrometry. Antifungal activity was determined by the dry paper disk and minimum inhibitory concentration (MIC) methods and identified by scanning electron microscopy and confocal microscopy. The results showed that PCA inhibited mycelial growth, where MIC was 25 µg mL-1. Microscopic analysis revealed a reduction in exopolysaccharide (EPS) formation, showing distorted and damaged hyphae of B. cinerea. The results suggested that PCA has a high potential in the control of B. cinerea and inhibition of EPS (important virulence factor). This natural compound is a potential alternative to postharvest control of gray mold disease.

Pharmacognostic evaluation, and development and validation of a HPLC-DAD technique for gallocatechin and epigallocatechin in rhizomes from Limonium brasiliense

ABSTRACT Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, is a plant from the southern coast of Brazilian that is used for the treatment of premenstrual syndrome, menstrual disorders and genito-urinary infections. The aim of the present study was to determine the quality control parameters for rhizomes collected during different periods by pharmacopoeial and non-pharmacopoeial methods, and to develop and validate a HPLC-DAD method for quantitative control of marker substances. The measured parameters were: granulometric analysis (d50 = 0.21–0.48 mm), loss on drying (11.1–12.4%), total ash (4.9–5.7%), dry residue by extraction with acetone:water (7:3, v/v) (30.6–39.5%), total polyphenol content (8.5–15.8%), and chromatographic fingerprint by HPLC and TLC. Besides, the acetone:water (7:3, v/v) extraction solvent in combination with a turbo-extractor, yielded the crude extract with a significant increase in tannins (F4,20 = 37.0, p < 0.001). The antioxidant potential of the crude acetone:water (7:3, v/v) extract, as well as the ethyl acetate and water fractions obtained after the partition process was evaluated by DPPH and the results were, respectively: IC50 6.87, 5.91, and 6.92 µg/ml. The validation parameters for the HPLC-DAD method showed adequate specificity, precision and accuracy. The gallo- and epigallocatechin contents were, respectively, 0.8–2.7% and 1.2–2.2%. These data contribute to analysis of the pharmacognostic quality control of the commonly used part from this species.

Antibacterial activity of Limonium brasiliense (Baicuru) against multidrug-resistant bacteria using a statistical mixture design.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae) is commonly known as "baicuru" or "guaicuru" and preparations of its dried rhizomes have been popularly used in the treatment of premenstrual syndrome and menstrual disorder, and as an antiseptic in genito-urinary infections. This study evaluated the potential antibacterial activity of rhizome extracts against multidrug-resistant bacterial strains using statistical mixture design. MATERIALS AND METHODS: The statistical design of four components (water, methanol, acetone, and ethanol) produced 15 different extracts and also a confirmatory experiment, which was performed using water:acetone (3:7, v/v). The crude extracts and their ethyl-acetate fractions were tested against vancomycin-resistant Enterococcus faecium (VREfm), methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae, all of which have been implicated in hospital and community-acquired infections. The dry residue, total polyphenol, gallocatechin and epigallocatechin contents of the extracts were also tested and statistical analysis was applied in order to define the fit models to predict the result of each parameter for any mixture of components. The principal component and hierarchical clustering analyses (PCA and HCA) of chromatographic data, as well as mass spectrometry (MS) analysis were performanced to determine the main compounds present in the extracts. RESULTS: The Gram-positive bacteria were susceptible to inhibition of bacterial growth, in special the ethyl-acetate fraction of ternary extracts from water:acetone:ethanol and methanol:acetone:ethanol against, respectively, VREfm (MIC=19µg/mL) and MRSA (MIC=39µg/mL). On the other hand, moderate activity of the ethyl-acetate fractions from primary (except water), secondary and ternary extracts (MIC=625µg/mL) was noted against KPC. The quadratic and special cubic models were significant for polyphenols and gallocatechin contents, respectively. Fit models to dry residue and epigallocatechin contents were not possible. PCA and HCA of the chromatographic fingerprints were disturbed by displacement retention time of some peaks, but the ultraviolet spectra indicated the homogeneous presence of flavan-3-ols characteristic of tannins. The MS confirmed the presence of gallic acid, gallocatechin, and epigallocatechin in extracts, and suggested the presence of monomers and dimers of B- and A-type prodelphinidins gallate, as well as a methyl gallate. CONCLUSION: Our results showed the antibacterial potential of L. brasiliense extracts against multidrug-resistant Gram-positive bacteria, such as VREfm and MRSA. The statistical design was a important tool to evaluate the biological activity by optimized form. The presence of some phenolic compounds was also demonstrated in extracts.

Bioactive Organocopper Compound from Pseudomonas aeruginosa Inhibits the Growth of Xanthomonas citri subsp. citri.

Citrus canker is a very destructive disease of citrus species. The challenge is to find new compounds that show strong antibiotic activity and low toxicity to plants and the environment. The objectives of the present study were (1) to extract, purify and evaluate the secondary metabolites with antibiotic activity produced by Pseudomonas aeruginosa LV strain in vitro against Xanthomonas citri subsp. citri (strain 306), (2) to determine the potential of semi-purified secondary metabolites in foliar application to control citrus canker under greenhouse conditions, and (3) to identify antibiotic activity in orange leaf mesophyll infected with strain 306, by electron microscopy. Two pure bioactive compounds were isolated, an organocopper antibiotic compound (OAC) and phenazine-1-carboxamide. Phenazine-1-carboxamide did not show any antibiotic activity under the experimental conditions used in this study. The OAC showed a high level of antibiotic activity with a minimum inhibitory concentration of 0.12 µg mL(-1). In greenhouse tests for control of citrus canker in orange trees, the semi-purified fraction F3d reduced lesion formation by about 97%. The concentration used was 500 times lower than that for the recommended commercial copper-based product. Electron microscopy showed that F3d altered the exopolysaccharide matrix and caused cell lysis of the pathogen inside the citrus canker lesions. These results suggest that secondary metabolites produced by inducing P. aeruginosa LV strain have a high potential to be used as a bioproduct to control citrus canker.

Effect of a Metalloantibiotic Produced by Pseudomonas aeruginosa on Klebsiella pneumoniae Carbapenemase (KPC)-producing K. pneumoniae.

Multidrug-resistant organisms (MDRO) are a great problem in hospitals, where thousands of people are infected daily, with the occurrence of high mortality rates, especially in infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-producing Kpn). The challenge is to find new compounds that can control KPC producing-Kpn infections. The aim of this study was to evaluate the antibiotic activity of the F3d fraction produced by the Pseudomonas aeruginosa LV strain against clinical isolates of KPC-producing Kpn. The results showed that the minimum inhibitory concentration of F3d (62.5 µg mL(-1)), containing an organic metallic compound, killed planktonic cells of KPC-producing Kpn strains after 30 min of incubation. At the same concentration, this fraction also showed an inhibitory effect against biofilm of these bacteria after 24 h of incubation. Treatment with the F3d fraction caused pronounced morphological alterations in both planktonic and biofilm cells of the bacteria. The inhibitory effect of the F3d fraction seems to be more selective for the bacteria than the host cells, indicating its potential in the development of new drugs for the treatment of infections caused by KPC-producing Kpn and other MDRO.

Antifungal Activity of Condensed Tannins from Stryphnodendron adstringens: Effect on Candida tropicalis Growth and Adhesion Properties.

Candida species are some of the most common causes of fungal infection worldwide. The limited efficacy of clinically available antifungals warrants the search for new compounds for treating candidiasis. This study evaluated the effect of condensed tannin-rich fraction (F2 fraction) of Stryphnodendron adstringens on in vitro and in vivo growth of Candida tropicalis, and on yeast adhesion properties. F2 exhibited a fungistatic effect with the minimum inhibitory concentration ranging from 0.5 to 8.0 µg/mL. A significant reduction in biofilm mass was observed after either pretreatment of planktonic cells for 2 h (mean reduction of 46.31±8.17%) or incubation during biofilm formation (mean reduction of 28.44±13.38%) with 4x MIC of F2. Prior exposure of planktonic cells to this F2 concentration also significantly decreased yeast adherence on HEp-2 cells (mean reduction of 43.13±14.29%), cell surface hydrophobicity (mean reduction of 25.89±10.49%) and mRNA levels of the genes ALST1-3 (2.9-, 1.8- and 1.8-fold decrease, respectively). Tenebrio molitor larvae, which are susceptible to C. tropicalis infection, were used for in vivo testing. Treatment with 128 and 256 µg/mL F2 significantly increased the survival of infected larvae. These results indicate a combined effect of F2 on inhibition of yeast growth and interference in yeast adhesion, which may contribute to the suppression of infection caused by C. tropicalis, thus reinforcing the potential of the condensed tannins from S. adstringens for the development of novel antifungal agents.

Acute toxicity and long-term safety evaluation of the crude extract from rhizomes of Limonium brasiliense in mice and rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, popularly known as baicuru, has been used in folk medicine to treat menstrual cramps and to regulate menstrual periods. However, little is known about its safety. This study evaluated the safety through in vivo tests of the acute, long-term, and liver toxicity, and the mutagenicity of the crude extract (CE) from rhizomes of L. brasiliense. MATERIALS AND METHODS: The acute toxicity was assessed in Swiss mice, and the chronic toxicity in Wistar rats. Male and female mice received the CE orally in single doses of 1.0, 2.0, 3.0, 4.0, or 5.0 g/kg. Clinical changes and mortality rate were used as parameters to assess the acute toxicity. In the long-term evaluation, male and female Wistar rats were treated orally with daily doses of the CE (50, 100, or 200 mg/kg) for 90 days. Assessments of weight, behavior and food intake, urinalysis, biochemical and hematological analyses, as well as macro- and microscopic observations of several organs were performed. The redox state of the liver was evaluated as a means of investigating the liver toxicity, and the micronucleus test to assess mutagenicity was also performed. RESULTS: Evaluation of acute toxicity indicated no apparent clinical change in the animals; the LD50 was 4.8 g/kg. Evaluation after 90 days administration showed that the CE, even in higher doses than are considered therapeutic, appeared to be safe. The micronucleus test demonstrated a low mutagenic potential for the CE. CONCLUSION: Our results showed that treatment with the CE from L. brasiliense caused low or no toxicity, as assessed using these doses and evaluation methods.

Microparticles containing guaraná extract obtained by spray-drying technique: development and characterization

AbstractGuaraná (Paullinia cupana Kunth, Sapindaceae) is well known for its dietary and pharmaceutical potential, and the semipurified extract of guaraná shows antidepressant and panicolytic effects. However, the low solubility, bioavailability and stability of the semipurified extract limit its use as a component of pharmaceutical agents. Delivery of the semipurified extract in a microparticle form could help to optimize its stability. In this study, microparticles containing semipurified extract of guaraná were obtained by the spray-drying technique, using a combination of maltodextrin and gum arabic. The raw materials and microparticles produced were characterized by particle size analysis, differential scanning calorimetry, thermogravimetric analysis, and scanning electron microscopy. The drug content and antioxidant capacity were also evaluated. In vitrodissolution tests using flow cell dissolution apparatus, were carried out to investigate the influence of formulation parameters on the release of semipurified extract of guaraná from the microparticles. The spray-drying technique and the processing conditions selected gave satisfactory encapsulation efficiency (80–110%) and product yield (55–60%). The mean diameter of microparticles was around 4.5 µm. The DPPH radical scavenging capacity demonstrated that microparticles can protect the semipurified extract of guaraná from the effect of high temperatures during the process maintained the antioxidant capacity. Differential scanning calorimetry results indicated an interaction between semipurified extract of guaraná and gum arabic: maltodextrin in the microparticles, and thermogravimetric analysis indicate that the profile curves of the microparticles are similar to the adjuvants used in drying, probably due to the higher proportion of adjuvants compared to semipurified extract of guaraná. In vitro dissolution tests demonstrate that all formulations complete dissolution within 60 min. Microencapsulation improved the technological characteristics of the powders and preserved the antioxidant properties. The study demonstrated the feasibility of producing these microparticles for a one-step process using spray drying. The composition of each formulation influenced the physical and chemical characteristics. This spray-drying technique can be used as an efficient and economical approach to produce semipurified extract of guaraná microparticles.

Antiviral Activity of Trichilia catigua Bark Extracts for Herpesvirus and Poliovirus.

Herpesvirus and poliovirus are responsible for important diseases in human and animal. Trichilia catigua a Brazilian native plant known as catiguá has several medicinal properties among them antimicrobial for bacteria and protozoa, however, no antiviral activity has been reported yet. This study evaluated the antiviral activity of the crude extract (CE) and aqueous and ethyl acetate fractions (AF, EAF) obtained from T. catigua in the replication of the Herpes simplex virus (HSV-1), bovine herpesvirus (BoHV-1) and poliovirus (PV-1). The cytotoxicity was analyzed by MTT assay and the antiviral effect was determined by the addition of extracts (0.25 to 100.0 µg/ml), before (-2h and -1h), during (Oh) and after (1h and 2h) the viral infection, by plaque reduction assay, in HEp-2 cell culture. The virucidal activity and inhibition of viral adsorption were also evaluated. In addition, the combination index (CI) with Acyclovir (ACV - reference drug) was determined for HSV-1. CE, AF and EAF showed a low toxicity (CC(50) >400 µg/ml) and low inhibitory concentration (IC50), ranging from 2.44-34.25 µg/ml for herpesvirus and 0.67 to 1.8 µg/ml for PV-1, associated with high selectivity index. The tested compounds showed high virucidal effect and high ability to inhibit viral adsorption, for all virus. The CI demonstrated a synergic effect (CI<1) for AF and EAF comparatively to acyclovir (ACV). Our study demonstrated that the extract and fractions of T. catigua is promising for future antiviral drug design with economically feasible production.

Extraction of flavonoids from Tagetes patula: process optimization and screening for biological activity

The flowers of Tagetes patula L., Asteraceae, commonly known as French marigold, are used in folk medicine as an antiseptic, diuretic, blood purifier and insect repellent. This study was conducted to optimize the extraction process through the biomonitoring of flavonoids, using a statistical mixture simplex-centroid design, to evaluate the effect of the solvents water, ethanol and acetone, as well as mixtures of these solvents, assessed by the total flavonoid content. The extracts were tested for dry residue, radical scavenging activity, chromatographic profile, and larvicidal activity. The acetone extract had the highest total flavonoid content, 25.13 ± 1.02% (4.07%); and the best radical scavenging activity, with IC50 of 15.74 μg/ml ± 1.09 (6.92%), but with lower dry residue, 6.62 ± 1.33% (20.10%). The water extracts showed higher levels of dry residue, but lower total flavonoid content and radical scavenging activity than the acetone extract. The positive correlation between the total flavonoid content and radical scavenging activity of the extracts showed that flavonoids contribute significantly to the antioxidant capacity. The statistical mixture design allowed us to optimize the extraction of flavonoids from flowers of T. patula, with acetone as the best extraction solvent. Preliminary studies on the biological activity of the optimized extracts demonstrated a larvicidal effect of the acetone extract on Aedes aegypti mosquitoes.

Morphoanatomy and pharmacognostic study of the wood of Croton echioides, the Northeastern Marapuama

Croton echioides Baill., Euphorbiaceae, is a small tree found in Bahia, Northeastern Brazil. Its stem bark has been widely sold as an aphrodisiac and tonic, as a substitute for the roots of Ptychopetalum olacoides Benth. Olacaceae, the Amazon Muira Puama or Marapuama, and C. echioides is characterized as the "Northeastern Marapuama". This contribution describes a morphoanatomical analysis and pharmacognostic study of stem bark of this species. The stem has a thick cortex with compound starch grains and laticifers; a sclerenchymatic sheath which consists of brachysclereids with large crystals externally to the phloem, and abundant fiber in the secondary xylem, as the main features of the species. The data obtained for water content (9.26±0.07%), water-soluble extractives (3.92±0.19%), total ash (1.24±0.06%) and acid-insoluble ash (0.16±0.01%), together with the chromatographic profile obtained by TLC, contribute to the quality control and standardization for the plant drug. The pharmacological screening indicated LD50 values above 500 mg/kg orally and equal to 500 mg/kg by the i.p. route, as well as some stimulant potential, depending on the dose.

Quality control and TLC profile data on selected plant species commonly found in the Brazilian market

The use of thin-layer chromatography (TLC) is a commonplace practice and can be of significant help to different laboratories with quality control, especially those that work with plant extracts and phytotherapeutics. This study evaluated ten species of plants (Schinus terebinthifolius; Arctium lappa; Trichilia catigua; Camellia sinensis; Mikania glomerata; Croton echioides; Achyrocline satureioides; Heteropterys aphrodisiaca; Plantago major; Arctostaphylos uva-ursi) that are commonly sold by compounding pharmacies, using TLC with reference substances and pharmacopoeic physical and chemical tests (loss on drying, level of extractives, and total ash content). The results showed that the ten species showed losses on drying consonant with the literature. The level of extractives for two species and total ash for five species were also consonant with the literature, and those of the other species were established in this study. The semipurified extracts of the ten species were assayed by TLC, and the analysis with the use of reference substances proved to be effective, in addition to being practical, simple, versatile, and economically viable.

Antimicrobial, cytotoxic and antioxidant activities and determination of the total tannin content of bark extracts Endopleura uchi.

Endopleura uchi is a typical Amazonian tree and its bark is popularly employed in the preparation of teas against myomas, arthritis, influenza, diarrhea and cancer. In this study, the antioxidant activity, cytotoxicity and antimicrobial activity of five different extracts of the bark, selected by their total tannin content, were assessed. The potential antioxidant activity of the extracts was determined by 2.2-diphenyl-1-picrylhydrazyl radical scavenging assay and the values found were very similar among the extracts and to the standards antioxidants used in the tests. Cytotoxicity analysis in mammalian cells indicated that all the tested extracts exhibited IC(50) values higher than the highest concentration used, showing that they do not present a risk when consumed under these conditions. Extract tested against five bacterial strains and one yeast strain did not show satisfactory growth inhibitory activity, and even the extracts that showed some antimicrobial activity were not effective at any dilution to determine the minimum inhibitory concentration. The results may serve as a reference for subsequent works, since such reference values described in the literature for the bark of E. uchi.

Antimicrobial and cytotoxic activities of medicinal plants of the Brazilian cerrado, using Brazilian cachaça as extractor liquid.

ETHNOPHARMACOLOGICAL IMPORTANCE: Many species of plants in the Brazilian cerrado (savanna) are widely used in ethnomedicine. However, the safety and effectiveness of medicinal plants used in communities with little or no access to manufactured drugs should be evaluated. AIM OF THE STUDY: Evaluate the antimicrobial and cytotoxic activities of extracts from eight plant species, obtained using Brazilian cachaça as the extractor liquid. MATERIALS AND METHODS: The extracts were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Candida parapsilosis, promastigote forms of Leishmania amazonensis, and poliovirus. In addition, cytotoxic activity was assayed in Vero cells and in human erythrocytes. RESULTS: The plant species Curatella americana, Sclerolobium aureum, and Plathymenia reticulata showed the best activity against yeasts, especially the crude extract of C. americana and its ethyl-acetate fraction. Kielmeyera lathrophyton showed a minimum inhibitory concentration of 250 µg/ml against S. aureus, and was inactive against gram-negative bacteria. The extract obtained from Annona coriacea showed the best activity against the promastigote forms of Leishmania amazonensis (IC(50)=175 µg/ml). Only C. americana showed potential for antipoliovirus activity. The concentrations of the crude extracts that showed toxicity to VERO cells had CC(50) between 31 and 470 µg/ml, and the lyophilized Brazilian cachaça showed a CC(50) of 307 µg/ml. None of the extracts showed toxicity against human erythrocytes. CONCLUSIONS: Among the plant species studied, C. americana proved to be effective against microorganisms, especially as an antifungal. The results will help in the search for alternative drugs to be used in pharmacotherapy, and will contribute to establish safe and effective use of phytomedicines in the treatment of infectious diseases.

Controle farmacognóstico das raízes de Heteropteris aphrodisiaca O. Mach. (Malpighiaceae) / Pharmacognostic analysis of the roots of Heteropteris aphrodisiaca O. Mach. (Malpighiaceae)

As raízes de Heteropteris aphrodisiaca, denominadas de " nó-de-cachorro" , pela semelhança morfológica com o pênis canino, são usadas popularmente com ação afrodisíaca, estimulante e no tratamento de disenterias, tendo este estudo o objetivo de caracterizá-las morfoanatômica e fisico-quimicamente. Para as análises foram elaboradas lâminas histológicas e analisados extratos aquosos, hidro-etanólicos e cetônicos obtidos a partir de amostras coletadas no estado do Mato Grosso, nas diferentes estações do ano. Tais raízes são tuberosas, cilíndricas e recobertas por súber estriado de coloração marrom-escura. O córtex, composto por tecido parenquimático, não apresenta elementos esclerificados, mas mostra-se com abundância de drusas e cristais prismáticos de oxalato de cálcio, bem como de idioblastos contendo polifenóis. No xilema secundário são abundantes as fibras gelatinosas, típicas de raízes de espécies do cerrado brasileiro. As análises físico-químicas revelaram maiores perdas por dessecação nas amostras coletadas no verão e na primavera. A água foi considerada o melhor líquido extrator, o teor de cinzas totais das amostras variou de 3,4 por cento no verão a 5,3 por cento no outono, enquanto que os teores de polifenóis totais foram maiores na primavera (10,2 por cento) que nas demais estações, sugere-se esta estação como a mais apropriada para a coleta da droga vegetal. Análises cromatográficas em camada delgada e líquida de alta eficiência foram realizadas empregando-se marcadores flavonoídicos neoastilbina, astilbina e isoastilbina, com valores de Rf de 0,60, 0,68 e 0,74, e tempos de retenção de: 16,44, 16,91 e 18,08 min, respectivamente.

The roots of Heteropteris aphrodisiaca, called " nó-de-cachorro" (dog-knot) for their morphological similarity to the canine penis, are used in popular medicine as an aphrodisiac or stimulant, and to treat dysentery. The objective of the present study was to characterize these roots morphoanatomically and physico-chemically. For the analyses, histological slides were prepared, and aqueous, hydro-ethanol, and ketone extracts were analyzed, from root samples collected in the state of Mato Grosso, in different seasons of the year. The roots are tuberous, cylindrical, and covered with a striated suber. The cortex, composed of parenchymatous tissue, has no sclerified elements, but shows abundant druses and prismatic crystals of calcium oxalate, as well as idioblasts containing polyphenols. In the secondary xylem, gelatinous fibers are abundant, typical for the roots of species of the Brazilian cerrado (savanna). The physical and chemical analyses revealed greater losses through desiccation in the samples collected in spring and summer. Water was the best extraction liquid. Total ash content of the samples ranged from 3.4 percent in summer to 5.3 percent in autumn. Total phenol content was higher in spring (10.2 percent), suggesting this season as the most appropriate to harvest the plant drug. Chromatographic analyses through TLC and HPLC were employed using the flavonoids neoastilbin, astilbin, and isoastilbin as markers. The Rf values were 0.60, 0.68 and 0.74, and the retention time were 16.44, 16.91 and 18.08 min, respectively.

Controle da qualidade do fruto de Syzygium cumini (L. ) Skeels / Quality control of Syzygium cumini (L. ) Skeels fruits

Considerando-se a qualidade dos fitoterápicos, é importante salientar que a preocupação com esta questão inclui rigoroso acompanhamento das diferentes etapas do desenvolvimento e produção destes produtos, desde a coleta do vegetal até a disponibilidade do produto final. Neste trabalho foi realizado o controle da qualidade do fruto de Syzygium cumini (L.) Skeels, popularmente conhecido como jambolão. Para o controle físico-químico e microbiológico utilizaram-se metodologias farmacopéicas e não farmacopeicas. Os resultados obtidos mostraram que o fruto apresenta um teor de taninos de 4,2 por cento e a análise fitoquímica preliminar dos frutos de S. cumini evidenciou taninos, flavonóides, antocianidinas, iridóides, alcalóides e heterosídeos fenólicos simples. A análise microbiológica não apresentou crescimento de patógenos entre os outros testes realizados. Destaca-se a importância do estabelecimento de normas para o controle da qualidade para as plantas, a fim de que sejam utilizadas com finalidade fitoterápica.

Considering the quality of the phytotherapic agents, it is important to point out that the concern with this subject includes rigorous attendance of the different steps of the development and production of these products, from the collection of the vegetable to the availability of the final product. In this work the quality control of the Syzygium cumini (L.) Skeels fruit, popularly known as jambolão, was carried out. Pharmacopoeic and no pharmacopoeic methodologies were employed to physico-chemical and microbiological quality control. The obtained results showed that the fruits presents a content of tannins of 4.2 percent, the preliminary phytochemical analysis of the S. cumini fruits evidenced tannins, flavonoids, antocianidin, iridoids, alkaloids and heterosides simple phenols. The microbiological analysis didnÆt present phatogenic growth among the other accomplished tests. The work stands out the importance of the establishment of norms for the quality control for the plants, so that they are used with phytotherapic purpose.