Enhanced Skin Permeation of Punicalagin after Topical Application of Pluronic Micelles or Vesicles Loaded with Lafoensia pacari Extract.

Punicalagin, the principal ellagitannin of Lafoensia pacari leaves, has proven antioxidant activity, and standardized extracts of L. pacari can be topically used for skin aging management. We hypothesized that Pluronic nanomicelles or vesicles could solubilize sufficiently large amounts of the standardized extracts of L. pacari and provide chemical stability to punicalagin. The standardized extracts of L. pacari were obtained with an optimized extraction procedure, and the antioxidant activity was characterized. Formulations containing Pluronic at 25% and 35% were obtained with or without Span 80. They were characterized by average diameter, polydispersity index, punicalagin content, physicochemical stability, and rheology. A release and skin permeation study was carried out in vertical diffusion cells. The extraction procedure allowed quantifying high punicalagin content (i.e., 141.61 ± 3.87 mg/g). The standardized extracts of L. pacari showed antioxidant activity for all evaluated methods. Pluronic at 25 and Pluronic at 35 with standardized extracts of L. pacari showed an average diameter of about 25 nm. The addition of Span 80 significantly increased the mean diameter by 15-fold (p < 0.05), indicating the spontaneous formation of vesicles. Pluronic formulations significantly protected punicalagin from chemical degradation (p < 0.05). Pluronic at 25 formulations presented as free-flowing liquid-like systems, while Pluronic at 35 resulted in an increase of about 44-fold in |ƞ*|. The addition of Span 80 significantly reduced the Pluronic sol-gel transition temperature (p < 0.05), indicating the formation of vesicles. Formulations with Span 80 significantly enhanced punicalagin skin permeation compared to formulations without Span 80 (p < 0.05). Formulations with Span 80 were demonstrated to be the most promising formulations, as they allowed significant permeation of punicalagin (about 80 to 315 µg/cm2), which has been shown to have antioxidant activity.

Potentially pathogenic bacteria isolated from neglected air and surfaces in hospitals

Hospital infections (HI) are a serious public health problem in many countries. Several studies have identified strains correlating to surgical site infections, many with multi-resistance. The goals of this study was to quantify, to identify and to verify the resistance profile of microorganisms collected at two hospitals settings, and to alert health professionals how environmental contamination can influence hospital infection rates. For air sampling in operating rooms, intensive care unit and materials sterilization center, the impaction method (Spin Air, IUL®) and passive sedimentation were used. For the isolation of bacteria on surfaces and uniforms contact plates (RODAC®) were used. Identification of the microorganisms was performed using Vitek® 2 Systems. The antibiograms were conducted according to the disk diffusion method recommended by CLSI. The surgical center of hospital B presented more than 500 CFU/m3 in aerial microbial load. In the aerial microbiota of the sampled areas of both hospitals, M. luteus, S. haemolyticus and S. hominis spp hominis were the prevalent microorganisms, with a percentage greater than 30%. On the surfaces and uniforms there was a prevalence of M. luteus (40%) and S. hominis spp hominis (20%) among others, and some of the resistant strains were isolated from environments with microbial load within the recommended limits.

Risks associated with pathogenic fungi isolated from surgical centers, intensive care units, and materials sterilization center in hospitals. Risks associated with pathogenic fungi isolated from critical hospital areas.

The hospital environment requires special attention to air quality, since it needs to be healthy for the protection of patients and health professionals in order to prevent them against hospital infections. The objective of this study was to isolate, identify and evaluate the susceptibility profile of isolated fungi from two hospitals. For air sampling the impaction (Spin Air, IUL®) and passive sedimentation methods were used. For the isolation of fungi from surfaces, contact plates (RODAC®) were used. The identification of the fungi was performed by observing the macroscopic and microscopic aspects of the colonies, whereas for better visualization of fruiting structures, the microculture technique was performed on slides. To evaluate the susceptibility profile, the broth microdilution test recommended by CLSI was performed. Thirty-five isolates were identified: Aspergillus flavus (12), Aspergillus fumigatus (11), Aspergillus niger (1), Aspergillus terreus (2), Penicillium spp. (7), and Fusarium spp. (2) in the hospitals evaluated. All isolates had a minimum inhibitory concentration (MIC) more than 128 µg/ml for fluconazole; 0.5 to 4.0 µg/ml for amphotericin B (hospital 1), and all isolates from haospital 2 had MIC ≥2.0 µg/ml. In hospital 1, MIC for posaconazole ranged from 0.25 µg/ml to ≥32 µg/ml, and hospital 2 ranged from 0.5 to 1.0 µg/ml. The monitoring and evaluation of air quality and surfaces are essential measures for prevention and control of hospital infections, as these microorganisms are becoming increasingly resistant to antimicrobial agents, thus making treatment difficult, especially in immunocompromised individuals.

Antifungal potential of punicalagin against Cryptococcus neoformans species complex.

This study evaluated the antifungal activity and cytotoxicity profile of the ellagitannin punicalagin, a compound extracted from the L. pacari A. St.-Hil (Lythraceae) leaf, against Cryptococcus neoformans species complex. Minimum inhibitory concentrations (MIC) were checked using the broth microdilution method. Minimum fungicidal concentrations (MFC) and time of death were used to confirm the antifungal activity of the compound. The in vitro cytotoxicity of punicalagin was tested in BALB/c3T3 fibroblasts and A549 human lung cancer cell line, while the hemolytic potential was tested on sheep erythrocytes. The morphological changes induced in yeast strains by the presence of punicalagin were also analyzed. Tested on eight isolates of the C. neoformans complex punicalagin showed MIC of 0.5 to 4.0 µg/mL and MFC> 256 µg/mL. Punicalagin also demonstrated a good growth inhibitory activity in time-kill curves, but it was not able to achieve a statistically significant reduction of fungal growth suggesting a fungistatic effect of the compound. In vitro cytotoxicity studies using the two cell lines showed that punicalagin has low activity on these cells and no activity on sheep erythrocytes. Morphological changes were seen in the yeasts strains studied when treated with punicalagin. Therefore, punicalagin is a potential antifungal for important pathogenic yeasts and presents a low cytotoxicity profile associated with no hemolytic effects.

Obtaining a Dry Extract from the Mikania laevigata Leaves with Potential for Antiulcer Activity.

BACKGROUND: Mikania laevigata leaves are commonly used in Brazil as a medicinal plant. OBJECTIVE: To obtain hydroalcoholic dried extract by nebulization and evaluate its antiulcerogenic potential. MATERIALS AND METHODS: Plant material and hydroalcoholic extract were processed and analyzed for their physicochemical characteristics. A method using HPLC was validated to quantify coumarin and o-coumaric acid. Hydroalcoholic extract was spray dried and the powder obtained was characterized in terms of its physicochemical parameters and potential for antiulcerogenic activity. RESULTS: The analytical method proved to be selective, linear, precise, accurate, sensitive, and robust. M. laevigata spray dried extract was obtained using colloidal silicon dioxide as adjuvant and was shown to possess 1.83 ± 0.004% coumarin and 0.80 ± 0.012% o-coumaric acid. It showed significant antiulcer activity in a model of an indomethacin-induced gastric lesion in mice and also produced a gastroprotective effect. CONCLUSION: This dried extract from M. laevigata could be a promising intermediate phytopharmaceutical product. SUMMARY: Research and development of standardized dried extract of Mikania laevigata leaves obtained through spray drying and the production process was monitored by the chemical profile, physicochemical properties and potential for anti-ulcerogenic activity. Abbreviations used: DE: M. laevigata spray dried extract, HE: hydroalcoholic extract.

Chemopreventive effect and angiogenic activity of punicalagin isolated from leaves of Lafoensia pacari A. St.-Hil.

Punicalagin is the major ellagitannin constituent from leaves of Lafoensia pacari, a Brazilian medicinal plant widely used for the treatment of peptic ulcer and wound healing. Genotoxic, cytotoxic, antigenotoxic, and anticytotoxic effects of punicalagin were assessed using micronucleus (MN) test and comet assay in mice. Due to the extensive use of L. pacari in the wound healing process, we also assessed the angiogenic activity of punicalagin using the chick chorioallantoic membrane (CAM) angiogenic assay. The highest dose of punicalagin (50mg/kg) showed significant cytotoxic effect by MN test and in the co-treatment with cyclophosphamide (CPA), this cytotoxicity was enhanced. Co-treatment, pre-treatment and post-treatment of punicalagin with CPA led to a significant reduction in the number of DNA breaks and in the frequency of CPA-induced MN, indicating antigenotoxic effect. Using the CAM model, punicalagin exhibited angiogenic activity in all doses mainly at the lowest concentration (12.5µg/µL). Therefore, these findings indicate an effective chemopreventive role of punicalagin and a high capacity to induce DNA repair. Also, the angiogenic activity presented by punicalagin in this study could contribute for the processes of tissue repairing and wound healing.

An approach on phytoalexins: function, characterization and biosynthesis in plants of the family Poaceae / Uma abordagem sobre fitoalexinas: função, caracterização e biossíntese em plantas da família Poaceae

ABSTRACT: Phytoalexins are compounds that have been studied a few years ago, which present mainly antimicrobial activity. The plants of the family Poaceae are the most geographically widespread and stand out for their economic importance, once they are cereals used as staple food. This family stands out for having a variety of phytoalexins, which can be synthesized via the shikimic acid (the phenylpropanoids), or mevalonic acid, being considered terpenoid phytoalexins. The characterization of these compounds with antimicrobial activity is carried out using chromatographic techniques, and the high-performance liquid chromatography (HPLC) coupled with mass spectrometry are the most efficient methods in this process. This research aimed to present an approach of the function, characterization and biosynthesis of phytoalexins in plants of the family Poaceae.

RESUMO: As fitoalexinas são compostos que vêm sendo estudados há alguns anos, apresentando principalmente atividade antimicrobiana. As plantas da família Poaceae são as geograficamente mais difundidas e se destacam por sua importância econômica, uma vez que são os cereais que compõe a base da alimentação, em todos os continentes. Essa família se destaca por apresentar uma variedade de fitoalexinas, sendo que as mais importantes são sintetizadas a partir da via do ácido chiquímico. A caracterização desses compostos com atividade antimicrobiana é realizada por meio de técnicas cromatográficas, sendo que a cromatografia líquida de alta eficiência (HPLC) acoplada com espectrometria de massas é a técnica mais eficiente nesse processo. O trabalho apresenta como objetivo uma abordagem da função, caracterização, biossíntese e aplicações das fitoalexinas em plantas da família Poaceae.

Blocking the L-type Ca2+ channel (Cav 1.2) is the key mechanism for the vascular relaxing effect of Pterodon spp. and its isolated diterpene methyl-6α-acetoxy-7ß-hydroxyvouacapan-17ß-oate.

Pterodon spp. Vogel (Fabaceae), popularly known as "sucupira", has ethnopharmacological application which is described as having antispasmodic and relaxant effects. Hence, it was hypothesized that sucupira oil-resin (SOR) could induce smooth muscle relaxation. So, this study investigated the mechanisms involved in the vasorelaxant effect of SOR and its isolated diterpene (methyl-6α-acetoxy-7ß-hydroxyvouacapan-17ß-oate). Vascular reactivity experiments were performed using rat aortic rings (n=5-8) with (E+) or without endothelium (E-) in an isolated bath organ. The SOR (0-56 µg/mL) relaxed phenylephrine (E+: 86.7±7.1%; E-: 92.3±4.7%) and KCl contracted rings (E-: 97.1±2.8%). In the same way, diterpene (0-48 µg/mL) also relaxed phenylephrine (E+: 94.5±3.6%; E-: 92.2±3.4%) and KCl contracted rings (E-: 99.7±0.2%). The pre-incubation of arterial rings with cyclopiazonic acid (reticular Ca2+-ATPase inhibitor), tetraethylammonium (K+ channels blocker) or MDL-12,330A (adenylyl cyclesinhibitor) did not modify either SOR- or diterpeneinduced vasorelaxation. However, ODQ (guanylyl cyclase inhibitor) impaired only diterpene-induced vasorelaxation. SOR and diterpene significantly reduced CaCl2-induced contraction stimulated by Bay K8644 (1 µM), phenylephrine (0.1 µM) or KCl solution (40 mM). Computational molecular docking studies demonstrated that the vasodilator effect of diterpene relies on blocking the Cav 1.2 channel, and patch clamp results showed that diterpene substantially decreased the ionic current through Cav 1.2 in freshly dissociated vascular smooth muscle cells. These findings suggest that SOR and its isolated diterpene induce endothelium-independent vascular relaxation by blocking the L-type Ca2+ channel (Cav 1.2).

Microemulsion based on Pterodon emarginatus oil and its anti-inflammatory potential

This article reports the development of a pharmaceutical product containing vegetable actives from a Brazilian medicinal plant. The possibility of forming a microemulsion using Pterodon emarginatus ("sucupira") oil was evaluated and the anti-inflammatory potential of this microemulsion was also examined. A formulation was developed using P. emarginatus oil, a mixture of ethoxylated Castor Oil (Ultramone(r) R-540/propylene glycol 2:1) (surfactant/cosurfactant) and distilled water at a ratio of 10:15:75, respectively. The microemulsion which was selected was then subjected to the preliminary stability test and analyzed in terms of average diameter of droplets, pH, zeta potential, and polydispersity index, on the 1st, 7th, 15th, and 30th days after preparation and stored at different temperatures (5 ± 2 °C, 25 ± 2 °C, and 40 ± 2 °C). The anti-inflammatory in vivo activity of both oil and formulation were evaluated, using the experimental model of croton oil-induced ear edema. The preliminary stability test showed that the microemulsion stored at 5 and 25 °C retained its original features throughout the 30-day period. The anti-inflammatory potential of both oil and formulation was shown to be statistically significant (p < 0.001), when compared to the control group, however, the microemulsion proved to be more effective (p < 0.05) than the oil when applied directly to the ear.

Descrevemos o desenvolvimento de um produto farmacêutico contendo ativo vegetal baseado em uma planta medicinal brasileira. Foi avaliada a habilidade de formação de uma microemulsão à base do óleo de Pterodon emarginatus (sucupira) e seu potential anti-inflamatório. Uma formulação foi desenvolvida utilizando o óleo de P. emarginatus, mistura de óleo de mamona etoxilado (Ultramona(r) R-540)/propilenoglicol (2:1) (tensoativo/cotensoativo) e água destilada, na proporção de 10:15:75, respectivamente. A microemulsão selecionada foi submetida ao teste preliminar de estabilidade e foi analisada quanto ao diâmetro médio das gotículas, pH, potential zeta e índice de polidispersão, no 1º, 7º, 15º e 30º dias após o preparo e foram estocadas em diferentes temperaturas (5±2 °C, 25±2 °C e 40±2 °C). Avaliaram-se a atividade anti-inflamatória in vivo do óleo de sucupira e da formulação, usando o modelo experimental do edema de orelha induzido pelo óleo de cróton. No teste preliminar de estabilidade observou-se que a formulação estocada a 5 °C e a 25 °C mantiveram suas características originais durante 30 dias. O potencial anti-inflamatório de ambos, óleo de sucupira e formulação foi estatisticamente significativo (p <0.001), quando comparado ao controle, porém a microemulsão foi mais eficiente (p <0.05) que o óleo aplicado diretamente nas orelhas dos animais.

Composição química e atividade antimicrobiana dos óleos essenciais das folhas e flores de Callistemon viminalis (sol. ex Gaertn.) G. Don ex. Loudon (Myrtaceae) / Chemical composition and antimicrobial activity of essential oils of leaves and flowers of Callistemon viminalis (sol. ex Gaertn.) G. Don ex. Loudon (Myrtaceae)

O presente trabalho teve como objetivo determinar a composição química e avaliar a atividade antimicrobiana dos óleos essenciais obtidos das folhas (frescas e secas) e flores da Callistemon viminalis. Os óleos essenciais foram analisados por CG/EM. A concentração inibitória mínima (CIM) foi determinada utilizando o método de microdiluição em caldo frente a bactérias Gram-positivas e Gram-negativas. De um total de 96,72%, 98,70% dos compostos identificados nos óleos essenciais foram monoterpenos. O 1,8-cineol foi o componente majoritário no óleo essencial das folhas frescas (70,43%), folhas secas (71,71%) e flores (42,39%), seguido do tricicleno com 12,97% nas folhas frescas, 8,20% nas folhas secas e 28,08% nas flores. A CIM para todos os micro-organismos testados foi ≥ a 2.000 μg/mL. Esse estudo é o primeiro relato sobre a composição química e atividade antimicrobiana do óleo essencial das flores de C. viminalis.

The aim of this study was to determine the chemical composition and the antimicrobial activity of essential oils of Callistemon viminalis leaves (fresh and dried) and flowers. The essential oils were analyzed by GC/MS and the minimum inhibitory concentration (MIC) against Gram-positive and Gram-negative bacteria was determined by the broth microdilution method. Between 96.72% and 98.70% of the compounds identified in the essential oils were monoterpenes. 1,8-Cineole was the major component in the essential oils of fresh leaves (70.43%), dried leaves (71.71%) and flowers (42.39%), followed by tricyclene: 12.97% in the fresh leaves, 8.20% in the dried leaves and 28.08% in the flowers. The MIC for all microorganisms tested was ≥ 2,000 μg/mL, for all the essential oil samples. This study is the first report on the chemical composition and antimicrobial activity of essential oil from the flowers of C. viminalis.

Composição química e atividade antimicrobiana do óleo essencial e anatomia foliar e caulinar de Citrus limettioides Tanaka (Rutaceae) / Chemical Composition and Antimicrobial Activity of Essential Oil and Anatomy of Citrus limettioides Tanaka(Rutaceae) Leaves and Stem

Citrus limettioides (Rutaceae) é utilizada popularmente para tratamento da sinusite, e também como antitérmica e hipotensora. Este trabalho teve como objetivos: identificar os constituintes dos óleos essenciais da folha e da casca dos frutos de C. limettioides; avaliar a atividade antimicrobiana dos óleos essenciais da folha e da casca dos frutos e realizar o estudo anatômico da folha e caule jovem. Os óleos essenciais foram obtidos por hidrodestilação em aparelho de Clevenger modificado e analisados por CG-EM. A concentração inibitória mínima (CIM) foi determinada utilizando o método de microdiluição em caldo frente a bactérias Gram-positivas, Gram-negativas e fungos. Para o estudo anatômico foram realizados cortes histológicos a mão livre e as secções foram coradas com azul de Alcian/safranina, reagente de Steinmetz, lugol e analisados em microscopia de luz. O linalol foi o componente majoritário no óleo essencial das folhas (36,88 a 45,15%) e o limoneno no óleo da casca dos frutos (70,60 a 75,18%). O óleo essencial das folhas apresentou CIM de 2000 μg/mL frente às bactérias e nenhuma atividade frente aos fungos. O óleo essencial da casca dos frutos não apresentou atividade antimicrobiana. A folha é hipoestomática, predominando os estômatos anomocíticos; o mesofilo é dorsiventral com parênquima paliçádico bisseriado e parênquima lacunoso com 11 camadas de células. Cavidades secretoras foram observadas no mesofilo da folha, no pecíolo e no caule, enquanto cristais prismáticos foram identificados em todas as estruturas da folha e do caule. Os dados obtidos no estudo microscópico podem auxiliar na identificação taxonômica desta planta e na avaliação da autenticidade da droga vegetal.

Citrus limettioides (Rutaceae), a sweet lime, is popularly used to treat sinusitis, and as an antipyretic and hypotensive agent.the aim of this study was to identify the constituents of the essential oils from the leaf and fruit peel of C. limettioides, test the antimicrobial activity of these essential oils and perform an anatomical study of the leaf and young stem. Essential oils were isolated by steam distillation in a modified Clevenger apparatus and analyzed by GC-MS. the minimum inhibitory concentrations (MIC) against Gram-positive and Gram-negative bacteria and fungi were determined by broth microdilution.Anatomical studies were performed on freehand histological sections, which were stained with Alcian blue / safranin, Steinmetz reagent and lugol and analyzed by light microscopy. Linalol was the major component in the leaf essential oil (36.88 to 45.15%) and limonene in the fruit peel essential oil (70.60 to 75.18%).the essential oil from leaves showed antimicrobial activity against bacteria (MIC 2000 μg/mL) and no activity against fungi.the essential oil from the fruit peel showed no antimicrobial activity. the leaf is hypostomatic with predominance of anomocytic stomata, the mesophyll is dorsiventral with biseriate palisade and 11 layers of spongy parenchyma cells.Secretory cavities were observed among the mesophyll cells of the leaf, petiole and stem, whilst prismatic crystals were identified in all structures of leaf and stem. the anatomical data obtained in the microscopic study may help in the taxonomic identification of this plant and to verify the authenticity of the drug plant.

Chemical variability of the essential oils from fruits of Pterodon emarginatus in the Brazilian Cerrado

Pterodon emarginatus Vogel, Fabaceae, is a tree species commonly known as "sucupira-branca". It is a popular medicinal plant in the Brazilian cerrado (Savanna). This study investigates the chemical variability of the essential oils from fruits of P. emarginatus. The fruits were collected from five sites in the Brazilian Cerrado and their essential oils were analyzed by GC/MS. The results obtained by Principal Component and Cluster Analysis identified two groups: cluster I containing β-caryophyllene and δ-elemene and cluster II containing α-copaene, β-cubebene, allo-aromadendrene, α-cubebene and γ-muurulene. The Canonical Discriminant Analysis was used to differentiate between clusters on the basis of essential oil composition. The results showed high variability in the composition of the essential oils from fruits of P. emarginatus, contributing to studies of domestication of this species.

Hypotensive effect of Aspidosperma subincanum Mart. in rats and its mechanism of vasorelaxation in isolated arteries.

ETHNOPHARMACOLOGICAL RELEVANCE: Aspidosperma subincanum is a medicinal herb that is known to be useful for the treatment of cardiovascular-related illnesses. However, its effects and pharmacological mechanisms of action have not been studied. The aim of the present study was to determine the effect of an ethanol extract of Aspidosperma subincanum (EEAS) on blood pressure (in vivo) and vascular tension (in vitro) in the rat thoracic aorta. MATERIALS AND METHODS: Catheters were inserted into the right femoral vein and artery of anesthetized rats for EEAS infusion and the measurement of blood pressure, heart rate and aortic blood flow (flow probes were placed around the aorta). Moreover, the vasodilator effect of EEAS in isolated pre-contracted rat aortas was examined. RESULTS: Intravenous infusion of EEAS resulted in significant and dose-dependent hypotension, bradycardia and increased aortic blood flow. In isolated arteries, EEAS (0-27 µg/mL) induced a concentration-dependent relaxation of pre-contracted aortic rings; endothelial denudation potentiated this effect. Pre-treatment of the aortic rings with ODQ, an inhibitor of soluble guanylyl cyclase (sGC); MDL-12,330A, an inhibitor of adenylyl cyclase (AC); or CPA, a SERCA inhibitor, reduced EEAS-induced vasorelaxation. Treatment with an EEAS impaired contractions induced by phenylephrine (an adrenergic agonist) and Bay K 8644 (an L-type Ca(2+) channel activator). The blockade of K(+) channels with tetraethylammonium, clotrimazole, glibenclamide or 4-aminopyridine reduced the relaxation stimulated by EEAS. CONCLUSIONS: These findings suggest that EEAS induces hypotension associated with bradycardia. EEAS induces endothelium-independent vascular relaxation. The sGC/cGMP and AC/cAMP pathways, SERCA activation and Ca(2+) and K(+) flux across the sarcolemma, are likely involved in this relaxation.

Chemical Constituents of Essential Oil from Lippia sidoides Cham. (Verbenaceae) Leaves Cultivated in Hidrolândia, Goiás, Brazil.

Several studies involving the family Verbenaceae, occurring in the Brazilian Cerrado, have emphasized the popular use of many aromatic species. We highlight the use of Lippia sidoides Cham., known as "alecrim pimenta," native to northeastern Brazil and northern Minas Gerais. Leaves of this species were collected in antropized Brazilian Cerrado area, in Hidrolândia, Goiás, and their essential oils were extracted by hydrodistillation in a Clevenger-type apparatus and thereafter analyzed GC/MS. Among the compounds identified in this study were the most abundant oxygenated monoterpenes, followed by sesquiterpenes hydrocarbons. The oxygenated monoterpene 1,8-cineole was the major constituent followed by isoborneol and bornyl acetate. The chemical composition of essential oil described in this paper differs from that described in the literature for L. sidoides found in its native environment, where the major constituents are thymol and carvacrol.

Production and characterization of tyrosinase activity in Pycnoporus sanguineus CCT-4518 crude extract

Tyrosinase is an enzyme of industrial interest. The production and characterization of tyrosinase from P. sanguineus CCT-4518 were investigated. The selection of inductors, luminosity influence, inoculum size and type of culture medium on the production of tyrosinase and the effect of inhibitors on enzyme activity were performed. Optimum conditions for intracellular tyrosinase production was observed after 2 days using 0.15% L-tyrosine as inducer, in the presence of light, with inoculum size of 10 mycelium discs, using 2% malt extract broth medium, incubated at 30°C, and constant agitation of 150 rpm. Tyrosinase activity was completely inhibited by the addition of 6 mM salicylhydroxamic acid or phenylthiourea, however an inhibition of 4.15% was recorded by the addition of 0.1 mM sodium azide. No inhibition could be detected in case of 0.1 mM phenyl methanesulfonyl fluoride addition. Optimal conditions for intracellular tyrosinase activity using L-dopa as substrate were observed at pH 6.6 and 45°C. Thermal stability studies indicated that the enzyme is stable at 45°C for 15 minutes. Higher temperatures decreased tyrosinase activity. Enzyme production was confirmed by non-denaturing polyacrylamide gel electrophoresis and the protein profile was investigated by denaturing polyacrylamide gel electrophoresis.

Dissolution test of herbal medicines containing Paullinia cupana: validation of methods for quantification and assessment of dissolution / Ensaio de dissolução de medicamentos fitoterápicos contendo Paullinia cupana: validação de métodos de quantificação e avaliação de dissolução

"Guaraná" (Paullinia cupana) is used as a physical activity enhancer and stimulator due to its methylxanthines and condensed tannins. The aim of this work was to evaluate the dissolution behavior of five herbal medicines in the form of capsules and tablets containing guaraná. Assay and dissolution methods were validated and results obtained allowed simultaneous marker quantification with precision, accuracy, selectivity and robustness. Findings showed that 100 percent of the herbal medicinal products analyzed did not provide satisfactory results concerning the presence of four markers, 60 percent had three markers (caffeine, catechin and epicatechin), while 40 percent had only caffeine at tested dosage forms. In addition, after 30 minutes, only capsule A showed at least 80 percent of the dissolved markers. In other capsules, marker dissolution did not exceed 60 percent whereas 60 percent of the samples had some characteristic pharmacotechnical problems. These results evidence the need for rigorous quality control to help ensure the therapeutic action of these drugs. To this end, dissolution studies are an essential tool for quality assurance of herbal medicines.

Guaraná (Paullinia cupana) é utilizado como revigorante e estimulante devido à presença de metilxantinas e taninos condensados. Este trabalho visou avaliar o comportamento de dissolução de cinco fitoterápicos, na forma de cápsulas e comprimidos, contendo guaraná. O método de quantificação e de dissolução foram validados e os resultados obtidos permitiram a quantificação dos marcadores simultaneamente, com precisão, exatidão, seletividade e robustez. Foi verificado que 100 por cento dos fitoterápicos analisados encontravam em desacordo quanto à presença dos quatro marcadores, sendo que 60 por cento apresentaram três marcadores (cafeína, catequina e epicatequina) e 40 por cento apresentaram somente a cafeína. Além disso, após o tempo de 30 minutos de ensaio foi possível observar que somente a cápsula A apresentou pelo menos 80 por cento dos marcadores dissolvidos. Nas demais cápsulas o comportamento de dissolução apresentado pelos marcadores não ultrapassou 60 por cento e, além disso, 60 por cento das amostras apresentaram alguns problemas farmacotécnicos característicos. Frente aos resultados obtidos torna-se evidente a necessidade de um rigoroso controle de qualidade que contribua para assegurar a ação terapêutica desses medicamentos e, nesse sentido, o estudo de dissolução constitui uma ferramenta essencial para a garantia de qualidade dos fitoterápicos.

Impact of ultrasound-assisted extraction on quality and photostability of the Pothomorphe umbellata extracts.

Pothomorphe umbellata (L.) Miq. is a Brazilian shrub with therapeutic and economic applications. There are some reports on the technological development of P. umbellata preparations; however, there are no studies on the influence of non-conventional extraction procedures on the quality of P. umbellata extracts. The aim of this study was to evaluate the effects of the ultrasound-assisted extraction (UE) parameters upon the extraction yield (EY%) of 4-nerolidylcatechol (4-NC) and the antioxidant activity of P. umbellata extracts using a factorial design and response surface methodology. Extracts obtained by UE and percolation were compared, and the photostability of 4-NC was evaluated via the exposure of UVA and visible light to the samples. The most influential variables observed for the UE were the ethanol-to-water and drug-to-solvent ratios. UE improved the extraction kinetics of 4-NC from plant material and improved the antioxidant activity of the extracts. Some of the ultrasound extracts showed an antioxidant activity that was not proportional to their 4-NC concentration, which suggests the presence of other active antioxidant compounds in these P. umbellata extracts. There was no significant difference in the photostability of 4-NC between the percolated and ultrasound extracts. Surprisingly, the isolated 4-NC material was significantly more stable when exposed to UVA-visible light compared to 4-NC in the plant extracts.

Estudo da atividade antibacteriana de plantas medicinais, aromáticas e corantes naturais / Study of the antibacterial activity of medicinal plants, herbs and natural dyes

O emprego de recursos naturais como mecanismo auxiliar no tratamento de distintas patologias tem ressurgido, com ênfase, nos útimos tempos. Tendo em vista esta situação, este estudo teve por finalidade investigar a atividade antimicrobiana de algumas plantas medicinais e aromáticas bastantes comuns na cultura brasileira. Foi constatado que, de modo particular, o alecrim-pimenta (Lippia sidoides) levou a uma considerável inibição do crescimento bacteriano. Tambêm plantas como alecrim (Rosmarinus officinalis), cravo (Sysygium aromaticum), noz-moscada (Myristica fragrans), pimenta-da-jamaica (Pimenta dioica) e sálvia (Salvia officinalis) demonstraram constituir fontes eficientes de compostos bioativos antibacterianos.

Medicinal plants, natural colour and spices were examined in vitro for antibacterial effects on the growth of Escherichia coli (EPEC) serotype O:125, Salmonella thyphimurium, Staphylococcus aureus, Listeria monocytogenes ATCC 7644 and Yersinia enterocolitica ATCC 9610. The most effective medicinal plant was Lippia sidoides at concentration of 3 percent (w/v), that added to Trypticase Soya Broth completely inhibited the growth of S. thyphimurium, S. aureus, L. monocytogenes and Y. enterocolitica. Among the spices, the cloves (Sysygium aromaticum) extract had the strongest inhibitory activity on the growth of E. coli, S. thyphimurium e Y. enterocolitica demonstred through the agar difussion technique. Sage (Salvia officinalis) and rosemary (Rosmarinus officinalis) extracts inhibited the growth of L. monocytogenes and S. aureus. The nutmeg (Myristica fragrans) extract also prevented the development of L. monocytogenes.