Limonium brasiliense rhizomes extract against virulence factors of Porphyromonas gingivalis: Inhibition of gingipains, bacterial adhesion, and biofilm formation.

Periodontitis is clinically characterized by destruction of the tooth support system and tooth loss. Porphyromonas gingivalis (Pg) plays a dominant role in periodontitis. Fractions and isolated compounds from an acetone-water extract of the roots of Limonium brasiliense (Lb) were tested in vitro for their anti-adhesive capacity against Pg on human KB buccal cells, influence on gingipains, the main virulence factors of Pg, and biofilm formation. Fractions EAF and FLB7 (50 µg/mL) reduced the bacterial adhesion of Pg to KB cells significantly (63 resp. 70%). The proanthocyanidin samarangenin A inhibited the adhesion (72%, 30 µM), samarangenin B (71%, 20 µM), and the flavan-3-ol epigallocatechin-3-O-gallate (79%, 30 µM). Fraction AQF, representing hydrophilic compounds, reduced the proteolytic activity of Arginin-specific gingipain (IC50 12.78 µg/mL). Fractions EAF and FLB7, characterized by lipohilic constituents, inhibited Arg-gingipain (IC50 3 µg/mL). On Lysine-specific gingipain, AQF has an IC50 15.89, EAF 14.15, and FLB7 6 µg/mL. The reduced bacterial adhesion is due to a strong interaction of proanthocyanidins with gingipains. AQF, EAF, and FLB7 significantly inhibited biofilm formation: IC50 11.34 (AQF), 11.66 (EAF), and 12.09 µg/mL (FLB7). In silico analysis indicated, that the polyphenols act against specific targets of Pg, not affecting mammalian cells. Therefore, Lb might be effective for prevention of periodontal disease by influencing virulence factors of Pg.

Trichilia catigua against Helicobacter pylori: An in vitro, molecular and in silico approach.

Helicobacter pylori is a bacterium that is present in the stomach of about 50% of the global population and is associated with several gastric disorders, including cancer. Natural products with antimicrobial activity have been tested against H. pylori, among them Trichilia catigua (catuaba), which is widely distributed in Brazil. This study aimed to evaluate extracts of T. catigua bark against H. pylori via determination of the minimum inhibitory and bactericidal concentrations (MIC and MBC); evaluation of virulence factors by real-time PCR, synergism with standard antimicrobials and morphology by scanning electron microscopy and simulations of the mechanism of action by molecular docking. The ethyl acetate fraction provided the best results, with an MIC50 of 250 µg/mL and a 42.34% reduction in urease activity, along with reduced expression of the CagA and VacA genes, which encode for the main virulence factors. This fraction presented synergistic activity with clarithromycin, reducing the MIC of the drug by four-fold. Docking simulations suggested that the extracts inhibit fatty acid synthesis by the FAS-II system, causing damage to the cell membrane. Therefore, T. catigua extracts have potential as an adjuvant to treatment and are promising for the development of new anti-H. pylori drugs.

Delayed administration of Trichilia catigua A. Juss. Ethyl-acetate fraction after cerebral ischemia prevents spatial memory deficits, decreases oxidative stress, and impacts neural plasticity in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Trichilia catigua A. Juss (Meliaceae) is used in Brazilian folk medicine to alleviate fatigue and emotional stress and improve memory. Previous studies from our laboratory reported that an ethyl-acetate fraction (EAF) of T. catigua that was given before cerebral ischemia in vivo prevented memory loss and reduced oxidative stress and neuroinflammation. Despite the value of these findings of a neuroprotective effect of T. catigua, treatment that was given immediately before or immediately after ischemia limits its clinical relevance. Thus, unknown is whether T. catigua possesses a specific time window of efficacy (TWE) when administered postischemia. AIM OF THE STUDY: Given continuity to previous studies, we investigated whether an EAF of T. catigua maintains its neuroprotective properties if treatment begins at different time windows of efficacy after ischemia. We also evaluated, for the first time, whether T. catigua possesses neuroplasticity/neurotrophic properties. MATERIAL AND METHODS: Rats were subjected to transient global brain ischemia (TGCI) and then given a single dose of the EAF (400 mg/kg) or vehicle (1 ml/kg) orally 1, 4, or 6 h postischemia. The levels of protein PCG, GSH, and GSSG, and activity of SOD and CAT were assayed as markers of oxidative stress on the day after ischemia. In another experiment, naive rats underwent spatial learning training in a radial maze task and then subjected to TGCI. Delayed treatment with the EAF began 4 or 6 h later and continued for 7 days. Retrograde memory performance was assessed 10, 17, and 24 days postischemia. Afterward, brains were examined for neurodegeneration and neuronal dendritic morphology in the hippocampus and cerebral cortex. Another group received the EAF at 4 h of reperfusion, and 4 days later their brains were examined for GFAP and Iba-1 immunoreactivity. Lastly, ischemic rats received the EAF 4 h after ischemia and neural plasticity-related proteins, BDNF, SYN, PSD 95, and NeuN were measured in the hippocampus 7 and 14 days after ischemia. RESULTS: A single EAF administration 1, 4, or 6 h postischemia alleviated oxidative stress that was caused by ischemia, expressed as a reduction of the amount of the PCG and GSSG, normalization of the GSH/GSSG ratio, and the restoration of SOD activity. Ischemia caused the persistent loss of memory (i.e., amnesia), an outcome that was consistently ameliorated by treatment with the EAF that was initiated 4 or 6 h postischemia. The 4 h delay in EAF treatment positively impacted dendritic morphology in neurons that survived ischemia. TGCI reduced BDNF, SYN, PSD-95, and NeuN protein levels in the hippocampus and cerebral cortex. The EAF normalized SYN and PSD-95 protein levels. Ischemia-induced neurodegeneration and glial cell activation were not prevented by EAF treatment. CONCLUSION: The present study corroborates prior data that demonstrated the neuroprotective potential of T. catigua and extends these data by showing that the delayed administration of EAF postischemia effectively prevented memory impairment and decreased oxidative stress, dendritic deterioration, and synaptic protein loss within a TWE that ranged from 1 to 6 h. This specific TWE in preclinical research may have clinical relevance by suggesting the possible utility of this plant for the development of neuroprotective strategies in the setting of ischemic brain diseases. Another innovative finding of the present study was the possible neurotrophic/neuroplastic properties of T. catigua.

Teores de polifenóis totais e de umidade em amostras de rizomas de Limonium brasiliense / Total polyphenols and moisture contents in samples of Limonium brasiliense rhizomes / Polifenoles totales y contenido de humedad en muestras de rizomas de Limonium brasiliense

O Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae), chamado popularmente de baicuru ou guaicuru é uma erva perene presente em todo litoral sul do Brasil, sendo indicada tradicionalmente para síndrome pré-menstrual e distúrbios menstruais. Essas atividades são atribuídas à presença dos polifenóis concentrados em seus rizomas, que também apresentam atividades bacteriostáticas, antioxidantes, antivirais e anti-inflamatórias. No desenvolvimento de formulações feitas a partir da droga vegetal são necessários testes de controle de qualidade durante todo o processo a fim de garantir a segurança, eficácia e qualidade da matéria-prima até sua formulação final. As metodologias utilizadas no desenvolvimento deste trabalho foram de análises físico-químicas para o controle de qualidade, análises quantitativas para determinação do teor de polifenóis totais e, por fim, uma análise comparativa entre os resultados obtidos no ano da coleta do material vegetal (2010) e das mesmas características após 13 anos da coleta, utilizando amostras do L. brasiliense com a presença de fungos ou sem nos rizomas. Sendo assim, este trabalho teve como objetivo realizar o controle de qualidade comparativo dos rizomas de Limonium brasiliense coletados em 2010 em relação ao teor de polifenóis totais e de umidade em 2023. Os resultados obtidos evidenciaram a produção de polifenóis pelos fungos na amostra contaminada, além de mostrarem a qualidade de armazenagem da droga vegetal. Com isso, foi possível concluir que o teor de polifenóis totais da amostra com fungo aumentou após 13 anos da coleta, e que neste interim não houve mudanças no teor de umidade da amostra sem fungo, garantindo sua qualidade no desenvolvimento de formulações ou pesquisas futuras.

Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae), popularly called baicuru or guaicuru, is a perennial herb present throughout the southern coast of Brazil, traditionally indicated for premenstrual syndrome and menstrual disorders. These activities are attributed to the presence of polyphenols concentrated in its rhizomes, which also have bacteriostatic, antioxidant, antiviral and anti-inflammatory activities. When developing formulations made from plant-based drugs, quality control tests are necessary throughout the process to ensure the safety, efficacy and quality of the raw material until its final formulation. The methodologies used in the development of this work were physical-chemical analyzes for quality control, quantitative analyzes to determine the total polyphenol content and, finally, a comparative analysis between the results obtained in the year the plant material was collected (2010) and the same characteristics after 13 years of collection, using samples of L. brasiliense with or without the presence of fungi in the rhizomes. Therefore, this work aimed to carry out comparative quality control of Limonium brasiliense rhizomes collected in 2010 in relation to the content of total polyphenols and moisture in 2023. The results obtained showed the production of polyphenols by fungi in the contaminated sample, in addition to show the storage quality of the plant drug. With this, it was possible to conclude that the total polyphenol content of the sample with fungus increased after 13 years of collection, and that in the meantime there were no changes in the moisture content of the sample without fungus, guaranteeing its quality in the development of formulations or future research.

Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae), popularmente llamada baicuru o guaicuru, es una hierba perenne presente en toda la costa sur de Brasil, tradicionalmente indicada para el síndrome premenstrual y trastornos menstruales. Estas actividades se atribuyen a la presencia de polifenoles concentrados en sus rizomas, que también tienen actividades bacteriostáticas, antioxidantes, antivirales y antiinflamatorias. Cuando se desarrollan formulaciones elaboradas a partir de medicamentos de origen vegetal, son necesarios ensayos de control de calidad durante todo el proceso para garantizar la seguridad, eficacia y calidad de la materia prima hasta su formulación final. Las metodologías utilizadas en el desarrollo de este trabajo fueron análisis físico- químicos para control de calidad, análisis cuantitativos para determinar el contenido de polifenoles totales y, finalmente, un análisis comparativo entre los resultados obtenidos en el año de recolección del material vegetal (2010) y el mismas características después de 13 años de colecta, utilizando muestras de L. brasiliense con o sin presencia de hongos en los rizomas. Por lo tanto, este trabajo tuvo como objetivo realizar un control de calidad comparativo de los rizomas de Limonium brasiliense recolectados en 2010 en relación al contenido de polifenoles totales y humedad en 2023. Los resultados obtenidos mostraron la producción de polifenoles por hongos en la muestra contaminada, además de mostrar la calidad de almacenamiento de la droga vegetal. Con esto se pudo concluir que el contenido total de polifenoles de la muestra con hongos aumentó después de 13 años de colecta, y que mientras tanto no hubo cambios en el contenido de humedad de la muestra sin hongos, garantizando su calidad en el desarrollo. de formulaciones o investigaciones futuras.

Relationship between the toxicity of raw Annona muricata leaves extract and 7, 12-dimethylbenzathracene (DMBA) with histopathological changes in wistar rats / Relación entre la toxicidad del extracto de hojas crudas de Annona muricata y el 7, 12-dimetilbenz antraceno (DMBA) con cambios histopatológicos en ratas Wistar

Abstract Aims: To identify the histopathological alterations in organs of Wistar rats to evaluate toxic effects of use of Annonamuricata Raw Leaf Extract (AMRLE) alone or in association with DMBA. Settings and Design: Sixty female Wistar rats were used, separated into groups and treated with a single dose of 65 mg/kg of DMBA and/or with 50; 100 and 200 mg/kg of AMRLE. Hematoxylin-Eosin (HE) and 1% methylene blue stains were used in the histopathological analysis and quantification of Aberrant Crypts (ACs) and Aberrant Crypt Focus (ACF). Fischer and Kruskal Wallis tests were used in the statistical analysis. Results: The administration of 65 mg/kg of DMBA and/or 50, 100 and 200 mg/kg of AMRLE did not influence weight development. Some histopathological alterations (hepatic steatosis; inflammatory foci in the liver, kidney and lung; pulmonary lymphoid hyperplasia, ectasia and hyperplasia in mammary gland epithelium) and the development of ACs and ACF in the intestinal colon were observed in all groups, except in the group negative control, with no statistical difference between analysed groups. Conclusions: Histopathological alterations and the formation of ACs and ACF did not show a statistically significant difference between the groups analysed. However, although AMRLE has antioxidant effects due to the presence of phenolic components, there was still the formation of some pathological processes that may be related to the isolated toxic action of DMBA and/or associated with other components of AMRLE, since these changes were not seen in the negative control group.

Resumen Objetivos: Identificar las alteraciones histopatológicas en órganos de ratas Wistar para evaluar los efectos tóxicos del uso del Extracto de Hoja Cruda de Annona muricata (AMRLE) solo o en asociación con DMBA. Configuración y diseño: Se utilizaron sesenta ratas hembras Wistar, se separaron en grupos y se trataron con una dosis única de 65 mg/kg de DMBA y/o con 50, 100 y 200 mg/kg de AMRLE. Se utilizaron tinciones de hematoxilina-eosina (HE) y azul de metileno al 1% en el análisis histopatológico y la cuantificación de criptas aberrantes (CA) y focus de criptas aberrantes (FCA). En el análisis estadístico se utilizaron las pruebas de Fischer y Kruskal Wallis. Resultados: La administración de 65 mg/kg de DMBA y/o 50, 100 y 200 mg/kg de AMRLE no influyó en el desarrollo del peso. Se observaron algunas alteraciones histopatológicas (esteatosis hepática; focus inflamatórios en el hígado, riñón y pulmón; hiperplasia, ectasia en epitelio de la glándula mamaria e hiperplasia linfoide pulmonar) y el desarrollo de CA y FCA en el colon intestinal en todos los grupos, excepto en el grupo control negativo, sin diferencias estadísticas entre los grupos analizados. Conclusiones: Las alteraciones histopatológicas y la formación de CA y FCA no mostraron diferencias estadísticamente significativas entre los grupos analizados. Sin embargo, aunque AMRLE tiene efectos antioxidantes debido a la presencia de componentes fenólicos, aún existe la formación de algunos procesos patológicos que pueden estar relacionados con la acción tóxica aislada del DMBA y/o asociados con otros componentes de AMRLE, ya que estos cambios no fueron observados en el grupo control negativo.

Development and evaluation of topical formulations that contain hydroethanolic extract from Schinus terebinthifolia against HSV-1 infection

Abstract The therapeutic drugs to treat Herpes simplex virus (HSV) infections have toxic side effects and there has been an emergence of drug-resistant strains. Therefore, the search for new treatments for HSV infections is mounting. In the present study, semi-solid formulations containing a crude hydroethanolic extract (CHE) from Schinus terebinthifolia were developed. Skin irritation, cutaneous permeation, and in vivo therapeutic efficacy of the formulations were investigated. Treatment with the ointment formulations did not result in any signs of skin irritation while the emulsions increased the thickness of the epidermis in Swiss mice. The cutaneous permeation test indicated that the CHE incorporated in the formulations permeated through the skin layers and was present in the epidermis and dermis even 3 h after topical application. In vivo antiviral activity in BALB/c mice treated with the CHE ointments was better than those treated with the CHE emulsions and did not significantly differ from an acyclovir-treated group. Taken together, this suggests that the incorporation of CHE in the ointment may be a potential candidate for the alternative topical treatment of herpetic lesions.

In vitro antifungal activity and phytochemical characterization of Eugenia uniflora, Libidibia ferrea and Psidium guajava

The aim of this study was evaluate the in vitro antifungal activity of crude extracts from Eugenia uniflora, Libidibia ferrea and Psidium guajava. The extracts were obtained by turbo-extraction using water (AQ) or acetone-water (AC-W) (7:3, v/v) as solvents and lyophilized to obtain the crude extracts (CE). The CE were characterized by UV-Vis, TLC and HPLC. The activity of CEs was investigated against clinical isolates of Candida spp. and the Minimum Inhibitory Concentration (MIC), MIC50 and MIC90 were determinated. The analysis by TLC showed that all CEs presented polyphenols (flavonoids and tannins). The CEs from E. uniflora showed higher amount of polyphenols (30.35 ± 2.15%, AC-W) and the HPLC analysis revealed the tannins in all extracts. The CEs of E. uniflora showed MIC range from 1.9 to 500.0 µg/mL, and lower values of MIC50 and MIC90 against non-albicans Candida isolates. Regarding L. ferrea and P. guajava, the results showing MIC from 3.9 to 1000.0 µg/mL (CE-AQ) against C. albicans. The results demonstrate antifungal performance from CE against various species of Candida spp., suggesting that the herbal species may be use as new potential antifungal agents. Additionally, the polyphenol content can play a pivotal role in the antifungal properties of CE.

Simultaneous liquid chromatography- tandem mass spectrometry method to quantify epicatechin and procyanidin B2 in rat plasma after oral administration of Trichilia catigua (catuaba) extract and its application to a pharmacokinetic study

Abstract Trichilia catigua A. Juss., Meliaceae, known as catuaba in Brazil, is traditionally used for the treatment of stress, sexual impotence and memory deficits. To our knowledge, there is no analytical method described in literature for simultaneous quantification of catuaba extract marker substances in biological matrices. The aim of this study was to develop and validate a bioanalytical method by LC-MS/MS to quantify epicatechin and procyanidin B2 in rat plasma after administration of standardized extract of T. catigua. Chromatographic separation was achieved with a C18 column, methanol and 0.1% aqueous formic acid at a flow rate of 0.25 ml/min. Detection was performed using electrospray ionization in negative mode. The lower limits of quantification were 5 ng/ml and 12.5 ng/ml for procyanidin B2 and epicatechin, respectively. Intra- and inter-day assays variability were less than 15%. The extraction recovery was 104% for epicatechin and 74% for procyanidin B2 using one-step liquid-liquid extraction with ethyl acetate. Epicatechin and PB2 were detected in plasma up to 300 min after oral administration of 400 mg/kg of standardized extract of T. catigua in rats. This rapid and sensitive method for the analysis of the epicatechin and procyanidin B2 in rat plasma can be applied to pharmacokinetic studies.

In vitro evaluation of the protective effects of plant extracts against amyloid-beta peptide-induced toxicity in human neuroblastoma SH-SY5Y cells.

Alzheimer's disease (AD) is the most common form of dementia and has no cure. Therapeutic strategies focusing on the reduction of oxidative stress, modulation of amyloid-beta (Aß) toxicity and inhibition of tau protein hyperphosphorylation are warranted to avoid the development and progression of AD. The aim of this study was to screen the crude extracts (CEs) and ethyl-acetate fractions (EAFs) of Guazuma ulmifolia, Limonium brasiliense, Paullinia cupana, Poincianella pluviosa, Stryphnodendron adstringens and Trichilia catigua using preliminary in vitro bioassays (acetylcholinesterase inhibition, antioxidant activity and total polyphenol content) to select extracts/fractions and assess their protective effects against Aß25-35 toxicity in SH-SY5Y cells. The effect of the EAF of S. adstringens on mitochondrial membrane potential, lipid peroxidation, superoxide production and mRNA expression of 10 genes related to AD was also evaluated and the electropherogram fingerprints of EAFs were established by capillary electrophoresis. Chemometric tools were used to correlate the in vitro activities of the samples with their potential to be evaluated against AD and to divide extracts/fractions into four clusters. Pretreatment with the EAFs grouped in cluster 1 (S. adstringens, P. pluviosa and L. brasiliense) protected SH-SY5Y cells from Aß25-35-induced toxicity. The EAF of S. adstringens at 15.62 µg/mL was able completely to inhibit the mitochondrial depolarization (69%), superoxide production (49%) and Aß25-35-induced lipid peroxidation (35%). With respect to mRNA expression, the EAF of S. adstringens also prevented the MAPT mRNA overexpression (expression ratio of 2.387x) induced by Aß25-35, which may be related to tau protein hyperphosphorylation. This is the first time that the neuroprotective effects of these fractions have been demonstrated and that the electropherogram fingerprints for the EAFs of G. ulmifolia, L. brasiliense, P. cupana, P. pluviosa and S. adstringens have been established. The study expands knowledge of the in vitro protective effects and quality control of the evaluated fractions.

Paullinia cupana: a multipurpose plant - a review

Abstract Seeds of guarana (Paullinia cupana Kunth, Sapindaceae) feature diverse pharmacological functions, for example, antimicrobial, antioxidant, anticarcinogenic, stimulating, and cognitive functions, as well as liver protection and weight loss. Many of these actions are probably due to the high content of methylxanthines and tannins in its seeds. In Brazil, the world's largest producer of guarana, the plant material is predominantly used in the soft drinks industry, although it is also used in the cosmetic and pharmaceutical industries. Although the Amazon region has the largest cropping area, the state of Bahia is the main guarana producer in Brazil (71%). This review focuses mainly on the possible pharmacological actions of guarana that have been investigated. Moreover, it discusses less-considered topics, such as the toxicology and quality control of seeds and extractives of guarana that will ultimately influence the safety of its use. In addition, it presents a detailed discussion of the methods used to prepare herbal drugs and their extracts, focusing on the importance of standardization and on the direct impact of preparatory factors, on the pharmacological properties of guarana extracts.

Guaraná (Paullinia cupana) seeds: Selective supercritical extraction of phenolic compounds.

Approximately 70% of the Brazilian production of guaraná (Paullinia cupana) seeds is absorbed by the beverage industries. Guaraná has several pharmacological properties: energy stimulant, antimicrobial, chemoprophylactic, antigenotoxic, antidepressive, anxiolytic, and anti-amnesic effects. Supercritical carbon dioxide extraction of bioactive compounds from guaraná seeds was carried out and optimized by an orthogonal array design (OA9(3(4))). The factors/levels studied were: modifier(s) (ethanol and/or methanol), extraction time (20, 40, and 60min), temperature (40, 50, and 60°C), and pressure (100, 200, and 300bar). The statistical design was repeated with increasing proportions of modifiers. The percentage of modifier used was proportional to the amount of polar compounds extracted. The best conditions for the supercritical extraction, based on the content of polyphenols, epicatechin/catechin quantification, yield and operating cost, proved to be: 40% ethanol:methanol during 40min, under 40°C, and 100bar. The temperature had a significant effect on the total phenolic content.

Enhanced Cutaneous Wound Healing In Vivo by Standardized Crude Extract of Poincianella pluviosa.

Wound healing is a complex process that involves several biological events, and a delay in this process may cause economic and social problems for the patient. The search continues for new alternative treatments to aid healing, including the use of herbal medicines. Members of the genus Caesalpinia are used in traditional medicine to treat wounds. The related species Poincianella pluviosa (DC.) L.P. Queiroz increases the cell viability of keratinocytes and fibroblasts and stimulates the proliferation of keratinocytes in vitro. The crude extract (CE) from bark of P. pluviosa was evaluated in the wound-healing process in vivo, to validate the traditional use and the in vitro activity. Standardized CE was incorporated into a gel and applied on cutaneous wounds (TCEG) and compared with the formulation without CE (Control) for 4, 7, 10, or 14 days of treatment. The effects of the CE on wound re-epithelialization; cell proliferation; permeation, using photoacoustic spectroscopy (PAS); and proteins, including vascular endothelial growth factor (VEGF), superoxide dismutase 2 (SOD-2) and cyclooxygenase 2 (COX-2) were evaluated. The TCEG stimulated the migration of keratinocytes at day 4 and proliferation on the following days, with a high concentration of cells in metaphase at 7 days. Type I collagen formed more rapidly in the TCEG. PAS showed that the CE had permeated through the skin. TCEG stimulated VEGF at day 4 and SOD-2 and COX-2 at day 7. The results suggest that the CE promoted the regulation of proteins and helped to accelerate the processes involved in healing, promoting early angiogenesis. This led to an increase in the re-epithelialized surface, with significant mitotic activity. Maturation of collagen fibers was also enhanced, which may affect the resistance of the extracellular matrix. PAS indicated a correlation between the rate of diffusion and biological events during the healing process. The CE from P. pluviosa appears promising as an aid in healing.

Development of tablets containing semipurified extract of guaraná (Paullinia cupana)

This study evaluated the technological feasibility of producing a semipurified extract of guaraná (Paullinia cupana Kunth, Sapindaceae) in tablet form, using a direct-compression process. Maltodextrin and gum arabic were used to produce the extract microparticles, in order to protect the microparticles against such factors as temperature, oxidation, and humidity. Using pharmacopoeial methodologies, technological and physicochemistry tests (determination of residual moisture, of bulk and tapped density, Hausner ratio, compressibility and compactibility index, appearance, mean weight, hardness, friability, disintegration time, determination of EPA amount in tablets and in vitro release profile) were conducted. The formulation containing 200 mg of microparticles, 170 mg microcrystalline cellulose, and 10 mg lactose gave the best results in terms of hardness (116 N), friabilility (0.28%), mean weight (0.3821 g), and disintegration time (25 min) for a tablet designed for oral administration. The results met pharmacopoeial specifications, and the tablets are suitable for oral administration.

Avaliação do fator de proteção solar em fotoprotetores acrescidos com extratos da flora brasileira ricos em substâncias fenólicas / Evaluation of the Sun Protection Factor in sunscreens added to with vegetable extracts rich in phenolic substances

Sabe-se que o número de casos de câncer de pele tem crescido muito nos últimos anos. Com isso, novos produtos são constantemente desenvolvidos no intuito de aumentar a gama de substâncias para proteção solar. O principal objetivo desse trabalho foi a análise in vitro da propriedade fotoprotetora de uma formulação contendo filtros solares químicos, adicionada de extratos brutos de Guazuma ulmifolia, Maytenus ilicifolia, Stryphnodendron adstringens e Trichilia catigua, respectivamente, e a avaliação quanto ao acréscimo do Fator de Proteção Solar (FPS). O ensaio foi conduzido pelo método espectrofotométrico in vitro do Fator de Proteção Solar (FPS). O teor de Polifenóis Totais (PT) e sua correlação com a Capacidade Antioxidante (CA) dos extratos avaliados foram realizados. Os resultados demonstram que as formulações foram estáveis após a adição dos extratos, quanto a características físico-químicas (características macroscópicas, ensaio de centrifugação e pH) quando comparadas ao controle. O ensaio de FPS in vitro demonstrou decaimento do FPS das formulações acrescidas de extratos vegetais ricas em compostos fenólicos, exceto para o extrato de S. adstringens. Os FPS obtidos para os extratos foram ≤2. O teor em PT foram (%): G. ulmifolia, 24,26±0,34; M. ilicifolia, 14,66±0,18; S. adstringens, 34,38±0,62 e T. catigua, 41,60±0,13. Os valores da CA (IC50) frente ao radical DPPH foram (miug/mL): vitamina C (7,52±0,12), G. ulmifolia (8,94±0,13), M. ilicifolia (38,41±0,93), S. adstringens (7,31±0,15) e T. catigua (5,48±0,04). A CA foi diretamente proporcional ao teor de PT.

It is known that the number of cases of skin cancer has increased greatly in recent years. Therefore, new products are constantly developed in order to increase the range of substances for sun protection. The aim of this study was evaluate in vitro the property the sunscreen containing chemical sunscreens, with added to extracts of Guazuma ulmifolia, Maytenus ilicifolia, Stryphnodendron adstringens and Trichilia catigua, Sun Protection Factor (SPF). The test was conducted by the spectrophotometric method of in vitro Sun Protection Factor (SPF). The percentage of the Total Polyphenol (TP) and its correlation with Antioxidant Capacity (CA) of the extracts evaluated were performed. The results show that the formulations were stable after the addition of extracts, in relation as the physico-chemical characteristics (macroscopic characteristics, test centrifugation and pH) when compared to control (time 0). The FPS test in vitro showed decrease SPF of the formulations added of plant extracts rich in phenolic compounds, except for the extract of S. adstringens. The FPS for the extracts obtained were ≤2. The percentage of the PT were (%): G. ulmifolia, 24,26±0,34; M. ilicifolia, 14,66±0,18; S. adstringens, 34,38±0,62 e T. catigua, 41,60±0,13. The values of CA (IC50) against the DPPH radical were (miug/mL): vitamin C (7,52±0,12), G. ulmifolia (8,94±0,13), M. ilicifolia (38,41±0,93), S. adstringens (7,31±0,15) e T. catigua (5,48±0,04). CA was directly proportional to the content of PT.

Memory retrieval improvement by Heteropterys aphrodisiaca in aging rats

Few data exists about the pharmacological properties of Heteropterys aphrodisiaca O. Mach. (Malpighiaceae), which is native to the scrubland regions of Brazil. The present study investigated the effects of oral treatment with H. aphrodisiaca extract (BST0298) on the learning and memory of young (3-6 months) and aged (21-23 months) rats, and compared the in vitro antioxidant activity of three lots collected in different years. An improvement in the number of sessions to learn the task was observed in the left/right discrimination test in aged rats treated for 45 days with 25 mg/kg (7.0 ± 0.5; p=0.005) or 50 mg/kg (7.6 ± 0.6; p=0.012) compared with control old rats (11.0 ± 1.6). On the other hand, pre-treatment did not improve the performance of scopolamine-treated mice in the passive avoidance test. The in vitro malondialdehyde test showed that all three different extracts presented similar antioxidant activity. The flavonoids astilbin, isoastilbin and neoastilbin were isolated from the extract and may contribute to the biological activity. These results suggest that repeated treatment with H. aphrodisiaca improves learning and memory, probably by a non-muscarinic mechanism.

Existem poucos dados disponíveis sobre as propriedades farmacológicas da Heteropterys aphrodisiaca O. Mach. (Malpighiaceae), nativa da região do pantanal brasileiro. O presente estudo investigou o efeito do tratamento oral com um extrato de H. aphrodisiaca (BST0298) sobre a memória e aprendizagem de ratos jovens (3-6 meses) e idosos (21-23 meses) e comparou a atividade antioxidante in vitro de três lotes, coletados em diferentes anos. Melhora quanto ao número de sessões necessárias para aprender a tarefa foi observada no teste de discriminação direita/esquerda em ratos idosos tratados por 45 dias com doses de 25 mg/kg (7,0 ± 0,5; p=0,005) e 50 mg/kg (7,6 ± 0,6; p=0,012) comparados com ratos idosos controle (11,0 ± 1,6). Por outro lado, o pré-tratamento com o extrato não melhorou o desempenho de camundongos tratados com escopolamina no teste da esquiva passiva. Em relação à avaliação da atividade antioxidante in vitro pelo teste do malonodialdeído, os três lotes analisados apresentaram atividade antioxidante semelhante. Os flavonóides astilbina, isoastilbina e neoastilbina foram isolados do extrato e podem contribuir para a atividade biológica. Estes resultados sugerem que a administração repetida de H. aphrodisiaca melhora a memória e aprendizagem provavelmente por um mecanismo não muscarínico.

Development and validation of an HPLC method for the determination of epicatechin in Maytenus ilicifolia (Schrad.) Planch., Celastraceae / Desenvolvimento e validação de um método de CLAE para a determinação da epicatequina em Maytenus ilicifolia (Schrad.) Planch., Celastraceae

A simple, reproducible and efficient high-performance liquid chromatography (HPLC) method was developed. Water (0.05 percent TFA):acetonitrile (0.05 percent TFA) was used as the mobile phase in a gradient system for the determination of epicatechin (EP) in leaves of Maytenus ilicifolia (Schrad.) Planch. The analysis was performed using an RP C-18 column (5 µm) as the stationary phase, with a flow rate of 0.8 mL/min, at a wavelength of 210 nm for detection and determination. The main validation parameters of the method were also determined. The calibration curve was found to be linear, with a range of 10-120 µg/mL (EP). The correlation coefficient of the linear regression analysis was within 0.9988, and the detection and quantification limits were 28.61 and 86.77 µg/mL, respectively. The content of EP was successfully determined, with satisfactory reproducibility and recovery. Recovery of the EP was 99.32 percent. The method was successfully applied to the determination of epicatechin in leaves of M. ilicifolia. The interlaboratorial evaluation showed the reproducibility of the method with a relative standard deviation of 14.62 percent.

Um método simples, reprodutível e eficiente de cromatografia líquida de alta eficiência (CLAE) foi desenvolvido. Água (0,05 por cento TFA):acetonitrila (0,05 por cento TFA) foi utilizado como fase móvel em um sistema de gradiente para a determinação da epicatequina (EP) em folhas de Maytenus ilicifolia (Schrad.) Planch., Celastraceae. A análise foi realizada utilizando coluna RP C-18 (5 µm) como fase estacionária, com vazão de 0,8 mL/min, e comprimento de onda de 210 nm para a detecção e determinação. Os principais parâmetros de validação do método foram determinados. A curva analítica apresentou-se linear no intervalo de 10-120 µg/mL (EP). O coeficiente de correlação da análise de regressão linear foi de 0,9988, o limite de detecção e o limite de quantificação foram de 28,61 e 86,77 µg/mL, respectivamente. O conteúdo do EP foi determinado com sucesso, com boa reprodutibilidade e recuperação. Recuperação da EP foi 99,32 por cento. O método foi aplicado com sucesso na determinação da epicatequina em folhas de M. ilicifolia. A avaliação interlaboratorial demonstrou a reprodutibilidade do método com desvio padrão relativo de 14,62 por cento.

Wound-healing evaluation of ointment from Stryphnodendron adstringens (barbatimão) in rat skin / Avaliação da cicatrização da pomada de Stryphnodendron adstringens (barbatimão) na pele de rato

This study evaluated the cicatrizant effect of an ointment containing 1 percent of the ethyl-acetate fraction extracted from the stem bark of "barbatimão" (Stryphnodendron adstringens), in wounds made in the skin of rats, after 4, 7 and 10 days of treatment. Control wounds were treated with a base ointment without extract. The proliferation of keratinocytes in the area of reepithelialization was evaluated by counting the number of epithelial cells that were blocked in metaphase by vincristine sulfate. The length of the reepithelialized margin and the contraction of the wound were measured. Topical application of the "barbatimão" ointment stimulated proliferation of the keratinocytes, but had no effect on the length of the epithelium or on the contraction of the wounds.

Neste estudo, avaliou-se a atividade cicatrizante de uma pomada contendo uma fração acetato de etila 1 por cento obtida de cascas de "barbatimão" (Stryphnodendron adstringens) em feridas excisionais na pele de ratos após 4, 7 e 10 dias de tratamento. Feridas controle foram tratadas com pomada base, sem extrato. A proliferação dos queratinócitos na área reepitelizada foi avaliada através da contagem do número de queratinócitos bloqueados em metáfase, pelo sulfato de vincristina. O comprimento da margem reepitelizada e a contração das feridas foram mensurados. As feridas tratadas com barbatimão apresentaram um maior número de mitoses do que aquelas tratadas com a pomada base, em todos os tempos avaliados. A aplicação tópica da pomada de "barbatimão" estimulou a proliferação epitelial contudo não teve efeito sobre a migração dos queratinócitos ou sobre a contração das feridas.

Antioxidant capacity of extracts and isolated compounds from Stryphnodendron obovatum Benth / Capacidade antioxidante de extratos e compostos isolados de Stryphnodendron obovatum Benth

The extract from stem bark of Stryphnodendron obovatum Benth. was chromatographed on a Sephadex® LH-20 column, and yielded nine compounds: gallic acid (GA), p-hydroxybenzoic acid (PHB), gallocatechin (GC), epigallocatechin (EPG), 4'-O-methylgallocatechin (MGC), epigallocatechin-(4β→8)-epigallocatechin (EPEP), epigallocatechin-(4β→8)-gallocatechin (EPGC), robinetinidol-(4α→8)-gallocatechin (ROGC) and robinetinidol-(4β→8)-epigallocatechin (ROEP). Evaluation of the antioxidant capacity in vitro by the methods of DPPH free radical (IC50; μg/mL) and reduction of the phosphomolybdenum complex (RAC) gave the following results, respectively: crude extract 4.52 and 0.8242; ethyl-acetate fraction 4.04 and 0.9537; aqueous fraction 5.58 and 0.9275. The crude extract and ethyl-acetate fraction were shown to possess an antioxidant capacity comparable to that of vitamin C (4.93 and 1.0). The values obtained by the DPPH free-radical method for the isolated compounds were IC50 (μM): GA=8.89; PHB=10.12; GC=16.46; EPG=13.20; MGC=21.00; EPEP=6.89; EPGC=4.91; ROGC=7.78 and ROEP=6.20. Vitamin C and trolox showed 30.11 and 30.10, respectively. Dimers showed greater activity in scavenging free radicals, possibly related to the number of hydroxyls. However, compounds without a hydroxyl at position 5 of the A-ring (5-deoxy-proanthocyanidins) did not change the antioxidant activity of the DPPH free radical, as evaluated here for the first time. Among the monomers, there appeared to be a direct relationship in scavenging of free radicals because of the stereochemistry of the compounds. The presence of a methyl radical on the B-ring significantly reduced the scavenging of free radicals of gallocatechin. All compounds showed greater scavenging of radicals than vitamin C and trolox, and these two compounds showed no significant difference from each other.

O extrato das cascas de Stryphnodendron obovatum submetido à cromatografia em coluna, Sephadex® LH-20, forneceu nove substâncias: ácido gálico (GA), ácido p-hidróxibenzóico (PHB), galocatequina (GC), epigalocatequina (EPG), 4'-O-metilgalocatequina (MGC), epigalocatequina-(4β→8)-epigalocatequina (EPEP), epigalocatequina-(4β→8)-galocatequina (EPGC), robinetinidol-(4α→8)-galocatequina (ROGC) e robinetinidol-(4β→8)-epigalocatequina (ROEP). A capacidade antioxidante in vitro pelos métodos do radical livre DPPH (IC50; μg/mL) e do complexo fosfomolibdênio (CAR) apresentou os seguintes resultados, respectivamente: extrato bruto 4,52 e 0,8242; fração acetato de etila 4,04 e 0,9537 e fração aquosa 5,58 e 0,9275 demonstrando possuírem capacidade antioxidante quando comparados com vitamina C 4,93 e 1,0. Os valores obtidos pelo método do radical livre DPPH com as substâncias isoladas foram: IC50 (μM): GA=8,89; PHB=10,12; GC=16,46; EPG=13,20; MGC=21,00; EPEP=6,89; EPGC=4,91; ROGC=7,78 e ROEP=6,20. Vitamina C e trolox mostraram valores de 30,11 e 30,10, respectivamente. Os dímeros mostraram maior atividade no sequestro de radicais livres, possivelmente relacionada com o número de hidroxilas. No entanto, substâncias com ausência de hidroxila na posição 5 do anel A (5-desoxi-proantocianidinas) não alteraram o poder antioxidante frente ao radical livre DPPH, tendo sido avaliadas aqui pela primeira vez. Entre os monômeros, parece haver uma relação direta com a estereoquímica. A presença de um radical metila no anel B reduziu significativamente a atividade da galocatequina. Todas as substâncias isoladas mostraram maior atividade do que a vitamina C e trolox, e estes não mostraram diferença significativa entre si.

Determinação quantitativa de taninos em três espécies de stryphnodendron por cromatografia líquida de alta eficiência / Quantitative determination of tannins in three species of stryphnodendron by high performance liquid chromatography

Um método de separação e quantificação por cromatografia líquida de alta eficiência (CLAE) em fase reversa foi desenvolvido usando água (0,05 por cento de TFA):acetonitrila (0,05 por cento de TFA) como fase móvel, em sistema gradiente para a análise dos flavan-3-óis presentes em extrato semipurificado das cascas de Stryphnodendron adstringens, Stryphnodendron polyphyllum e Stryphnodendron obovatum. A CLAE foi realizada com a fração acetato de etila (FAE) sendo submetida à extração em fase sólida (cartucho C18-E) com metanol:água (2:8), filtrada por membrana de porosidade 0,5 μm; a pré-coluna e coluna empregadas foram Phenomenex® Gemini C-18 (5 μm), com esta última mantida a 30 ºC, com vazão de 0,8 mL/min e detecção a 210 nm. Utilizaram-se soluções dos padrões de ácido gálico e galocatequina para a obtenção da curva analítica. O método proposto foi validado de acordo com a resolução RE nº 899/2003 da ANVISA. A análise quantitativa da FAE das três espécies mostrou que existe similaridade no teor de galocatequina. S. adstringens possui ácido gálico em uma proporção superior a 60 por cento em relação às outras duas espécies. A metodologia desenvolvida mostrou-se viável à aplicação em plantas ricas em taninos, como nos casos de S. adstringens, S. polyphyllum e S. obovatum.

A method of separation and quantification by reverse-phase high-performance liquid chromatography (HPLC) was developed, using water (0.05 percent TFA):acetonitrile (0.05 percent TFA) as the mobile phase in a gradient system. Flavan-3-ols present in a semipurified extract from the stem bark of Stryphnodendron adstringens, Stryphnodendron polyphyllum and Stryphnodendron obovatum were analyzed. The HPLC was performed with the ethyl-acetate fraction (EAF) using a solid-phase extraction on cartridges C18-E with methanol:water (2:8), filtered through a membrane of 0.5 μm pore size; the column was Phenomenex® Gemini C-18 (5 μm) at 30 ºC, with a flow rate of 0.8 mL/min. The analysis was done at 210 nm. Gallic acid and gallocatechin solutions were used as calibration standards. The proposed method was validated by resolution RE No. 899/2003 of the National Health Surveillance Agency. Quantitative analysis of the EAF showed high contents of flavan-3-ols in the stem bark of all three species. This study demonstrated that it is possible to determine the concentration of individual substances in tannin-rich plants. The system developed can be used as a chromatographic profile for the semipurified fraction of S. adstringens, S. polyphyllum, and S. obovatum.