Evaluation of the anti-inflammatory effect of 1,4-dihydropyridine derivatives.

INTRODUCTION: Inflammation is a physiological event that protects the organism against different factors that lead to loss of tissue homeostasis. Dihydropyridine (DHP) derivatives are heterocyclic compounds known for their different biological activities, including anti-inflammatory activities. OBJECTIVE: To evaluate the anti-inflammatory activity of 1,4-dihydropyridine (1,4-DHP) derivatives using anti-inflammatory models in vitro, in RAW264.7 cells induced by lipopolysaccharide (LPS) and in vivo using the acute lung injury (ALI) model in mice. RESULTS: Fifteen compounds derived from 1,4-DHP were tested in RAW264.7 cells for their cytotoxic effect and cell viability. Thereafter, only the six compounds that showed the highest cell viability were tested for the production or inhibition of the pro-inflammatory cytokine interleukin 6 (IL-6). The best compound (compound 4) was tested for its anti-inflammatory effects in vitro and in vivo, showing inhibition of nitric oxide (NO), pro-inflammatory cytokines, increased phagocytic activity, and an increase in IL-10 in vitro. In in vivo tests, compound 4 also reduces the levels of NO, myeloperoxidase (MPO) activity, leukocyte migration, and exudation, as well as reducing the levels of tumor necrosis factor-alpha (TNF-α) and IL-6 and preventing the loss in the lung architecture. CONCLUSION: This compound showed important anti-inflammatory activity, with a significant ability to reduce the production of pro-inflammatory mediators and increase the phagocytic activity of macrophages and anti-inflammatory mediator secretion (IL-10). These findings led us to hypothesize that this compound can repolarize the macrophage response to an anti-inflammatory profile (M2). Moreover, it was also able to maintain its anti-inflammatory activity in vivo experiments.

Melanoma growth in non-chemically modified translucid bacterial nanocellulose hollow and compartimentalized spheres.

BACKGROUND: Bacterial nanocellulose (BNC) has been used as cell support in numerous tissue engineering studies. Its use can be explained based on the fact its structure allows the creation of a required microenvironment for an ideal material, which supports 3D cell culture. Its structure and interconnected pores lead to animal cells adhesion and proliferation, also allowing oxygen and nutrients transportation. METHODS: We developed a new methodology to produce spherical platforms synthesized by Komagataebacter hansenii (ATCC 23769) under dynamic culture conditions in minimal medium. The chemical composition and physical properties of the platforms were evaluated. Then, human melanoma cells (SK-MEL-28) were encapsulated into the platforms and evaluated by metabolic activity, morphology and their ability on adhering to the Hollow Translucid BNC Spheres (BNC-TS-H) and Compartmentalized Translucid BNC Spheres (BNC-TS-C) up to 3 days. RESULTS: BNC-TS-H and BNC-TS-C platforms were produced as translucid spheroid platforms with distinct microenvironment under dynamic fermentation. The chemical and physical characterizations confirmed the platforms composition as BNC. The produced internal microenvironments in spherical platforms are relevant to determine tumor cell fate. In the first 12 h of culture, cells could adhere to nanocellulose microfibers assuming their typical tumorous phenotype in 72 h of culture. CONCLUSION: The dynamic fermentation in minimal medium produced distinct microstructured platforms of BNC-TS-H and BNC-TS-C. The platforms microstructure resulted in microenvironments that enabled distinct cell-cell and cell-matrix interactions. This behavior suggests several applications in tissue engineering. GENERAL SIGNIFICANCE: The method produced translucid BNC sphere platforms with distinct microenvironments for 3D cell culture.

Anti-leukemic activity of semisynthetic derivatives of lupeol.

In our previous work, lupeol was isolated from aerial parts of V. scorpioides and modified by semisynthetic approach. The purpose of this study was to investigate the cytotoxicity of lupeol and its derivatives previously prepared on the human K562 acute myeloid leukemia cell and human Jurkat acute lymphoid leukemia cell in vitro. Compounds 3ß-hydroxylup-20(29)-en-30-al (2), lup-20(30)-en-3ß,29-diol (3), 3ß-acetoxylup-20(29)-en-30-al (5) and 3ß-acetoxy-30-hydroxylup-20(29)-ene (6) presented cytotoxicity with IC50 ranging from 11.72 to 56.15 µM at 24 h of incubation for both cell lines. Most of the active compounds (3, 5 and 6) were selective to leukemia cells, in compare with healthy cells. The hemolysis assay showed high blood compatibility of the cytotoxic lupeol derivatives which makes possible an intravenous administration of these compounds aiming to the potential to development of anti-leukemic drugs.

Metabolomic analysis among ten traditional "Arnica" (Asteraceae) from Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of several Asteraceae species in Brazil are popularly used as anti-inflammatory. Some of these species are popularly recognizes as "arnica" because of the morphological and sensorial analogy with the traditional European Arnica montana. These used species in Brazil were identified as Calea uniflora Less, Chaptalia nutans (L.) Polák, Lychnophora ericoides Mart. Lychnophora pinaster Mart. Lychnophora salicifolia Mart. Porophyllum ruderale (Jacq.) Cass, Pseudobrickellia brasiliensis (Spreng.) R. M. King & H. Rob. Sphagneticola trilobata (L.) Pruski and Solidago chilensis Meyen. However, the comparative chemical profile of these so-called "arnicas" has never been reported in the literature. AIM OF THE STUDY: This work aimed to compare the main plants recognized as "arnica" in Brazil by using metabolomic analysis, based on UPLC-ESI-QTof-MS2 data and multivariate statistical analysis. MATERIALS AND METHODS: The metabolites profiling of 10 "arnica" species were established by UPLC-ESI-QTof-MS2. Three tinctures of each species (dry leaves) were produced and one aliquot of each tincture was injected and analyzed three times by UPLC-ESI-QTof-MS2. Data were acquired both in negative and positive modes and processed by MassLynx®, MarkerLynx® and Matlab® softwares. Principal component analysis (PCA) was used to reduce dimensionality and data redundancy; hierarchical trees helped to identify and eliminate contaminated or misplaced injections/samples. To achieve the objectives both hierarchical and k-means clustering techniques were employed to group similar samples or species. RESULTS: Diagnostic analysis of MS data allowed the identification of 54 metabolites. The identification was supported with the use of an external standard, fragmentation pattern and data from the literature. The main classes of identified compounds included phenolic acids, coumarin, flavonoids, heterosides, terpenoids and nitrogen compounds. Cluster analysis revealed that Sphagneticola trilobata, Solidago chilensis and Lychnophora pinaster have some chemical features similar to those of Arnica montana. In contrast, the same statistical analysis also showed that Pseudobrickellia brasiliensis, Porophyllum ruderale and Chaptalia nutans are chemically diverse from Arnica montana. The variability of the samples relied principally on nitrogenated compounds (confidence level 4) found in P. brasiliensis and P. ruderale, three phenolic compounds (level 2) detected in P. brasiliensis and in C. nutans and triterpenes (level 3) found in L. salicifolia and L. pinaster. CONCLUSIONS: In summary, the mass spectrometry technique in conjunction with multivariate statistical analysis proved to be an excellent tool to identify correlated compounds, as well as to verify the chemical similarity among evaluated species. This methodology was successfully used to establish important correlations in medicinal preparations of so-called "arnicas" used in Brazil.

Construção do guia de plantas medicinais de Florianópolis, SC, Brasil / Production of the guide to medicinal plants in Florianópolis, Santa Catarina, Brazil / Construcción de la guía de plantas medicinales de Florianópolis, Estado de Santa Catarina, Brasil

Há escassez relativa de experiências com plantas medicinais e de orientações para confecção de materiais didáticos sobre elas na Atenção Primária à Saúde (APS). Uma equipe interinstitucional (Universidade Federal de Santa Catarina e Secretaria Municipal de Saúde de Florianópolis) produziu um guia de plantas medicinais em Florianópolis, SC. O objetivo deste trabalho é apresentar essa experiência e o guia, que é voltado para profissionais da APS e que valorizou saberes científicos, populares e profissionais, evitando uma abordagem colonizadora do tema. Após alguns tópicos introdutórios, o guia apresenta 38 plantas medicinais comumente usadas em Florianópolis (nomes científicos e populares; partes usadas; fotos; características botânicas; uso popular; informações científicas; uso clínico; modo de usar; reações adversas e contraindicações; plantas na gestação; toxicidade; glossário; e referências). Espera-se contribuir para a construção de outros guias adaptados aos contextos locais e para a valorização do uso de plantas medicinais na APS. (AU)

Experiences with medicinal plants and guidance for producing educational resources about these plants for use in primary health care (PHC) are relatively scarce. A team from the Santa Catarina Federal University and Florianópolis City Council Department of Health produced the "Guide to Medicinal Plants in Florianópolis". This article presents this experience and the guide, which is aimed at PHC professionals and values scientific, traditional and professional knowledge, avoiding a colonizing approach to the topic. After presenting some introductory topics, the guide describes 38 medicinal plants commonly used in Florianópolis (scientific and common names, parts used, photo, botanical characteristics, common uses, scientific information, clinical use, adverse reactions and contraindications, plants during pregnancy, toxicity, glossary and references). It is hoped this guide will contribute to the production of other guides adapted to local contexts and recognition of the value of medicinal plants in PHC. (AU)

Hay una escasez relativa de experiencias con plantas medicinales y de orientaciones para la confección de materiales didácticos sobre ellas en la atención primaria de la salud (APS). Un equipo interinstitucional (Universidad Federal de Santa Catarina y Secretaría Municipal de Salud de Florianópolis) produjo una guía de plantas medicinales en Florianópolis, estado de Santa Catarina. El objetivo de este trabajo es presentar esa experiencia y la guía, dirigida a los profesionales de la APS y que valorizó saberes científicos populares y profesionales, evitando un abordaje colonizador del tema. Después de algunos tópicos de introducción, la guía presenta 38 plantas medicinales comúnmente utilizadas en Florianópolis (nombres científicos y populares, partes usadas, foto, características botánicas, uso popular, informaciones científicas, uso clínico, modo de usar, reacciones adversas y contraindicaciones, plantas en la gestación, toxicidad, glosario y referencias). Se espera contribuir para la construcción de otras guías adaptadas a los contextos locales y para la valorización del uso de plantas medicinales en la APS. (AU)

Anti-Inflammatory Profile of Jungia sellowii Less. by Downregulation of Proinflammatory Mediators and Inhibition of NF-κB and p38 Pathways.

Jungia sellowii Less. (Asteraceae) is a native plant found in Southeast Brazil used traditionally to treat inflammatory diseases. This study was conducted (1) to investigate the toxicity of the crude extract (CE) and (2) to investigate the mechanism of the anti-inflammatory action of J. sellowii L. roots. The potential acute toxicity of CE was performed by administration of only different doses of CE (500, 1,000, and 2,000 i.p.) on mice for 14 days. The anti-inflammatory effect was evaluated using carrageenan-induced acute pleural cavity inflammation in a mouse model, evaluated through the following inflammatory variables: leukocyte, protein concentrations of the exudate, myeloperoxidase (MPO), adenosine deaminase (ADA), nitric oxide metabolites (NOx), and proinflammatory cytokine (tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin- (IL-) 6, and IL-12) levels in mouse pleural fluid leakage. The p65 protein phosphorylation of nuclear factor NF-kappa B (p65 NF-κB) and p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation were analyzed in lung tissue. Our results demonstrated that the administration of CE up to 2,000 mg/kg did not present a toxic effect. In addition, the pretreatment of mice with CE; its derived fractions (aqueous fraction (AqF), butanol fraction (BuOHF), and ethyl acetate fraction (EtOAcF)); and isolated compounds (curcuhydroquinone O-ß-glucose (CUR) and α and ß piptizol (Pip)) reduced the following inflammatory variables: neutrophils, protein concentrations of the exudate, MPO, ADA, NOx, and proinflammatory cytokine (TNF-α, IFN-γ, IL-6, and IL-12) levels in mouse pleural fluid leakage. The compounds CUR and Pip also decreased the p65 protein phosphorylation of NF-kappa B and p38 (MAPK) in lung tissue. J. sellowii L. has important anti-inflammatory activity with potential applications in drug development against inflammatory disorders. These effects found can be attributed to the ability of the new isolated compounds CUR and Pip to suppress p65 NF-κB and p-p38 MAPK pathways.

Systemic Administration of Calea pinnatifida Inhibits Inflammation Induced by Carrageenan in a Murine Model of Pulmonary Neutrophilia.

OBJECTIVE: The aim of this study was to investigate the anti-inflammatory effects of the crude extract (CE), derived fraction, and isolated compounds from Calea pinnatifida leaves in a mouse model of pulmonary neutrophilia. METHODS: The CE and derived fractions, hexane, ethyl acetate, and methanol, were obtained from C. pinnatifida leaves. The compounds 3,5- and 4,5-di-O-E-caffeoylquinic acids were isolated from the EtOAc fraction using chromatography and were identified using infrared spectroscopic data and nuclear magnetic resonance (1H and 13C NMR). Leukocytes count, protein concentration of the exudate, myeloperoxidase (MPO) and adenosine deaminase (ADA), and nitrate/nitrite (NO x ), tumor necrosis factor-alpha (TNF-α), interleukin-1-beta (IL-1ß), and interleukin-17A (IL-17A) levels were determined in the pleural fluid leakage after 4 h of pleurisy induction. We also analyzed the effects of isolated compounds on the phosphorylation of both p65 and p38 in the lung tissue. RESULTS: The CE, its fractions, and isolated compounds inhibited leukocyte activation, protein concentration of the exudate, and MPO, ADA, NO x , TNF-α, IL-1ß, and IL-17A levels. 3,5- and 4,5-di-O-E-caffeoylquinic acids also inhibited phosphorylation of both p65 and p38 (P < 0.05). CONCLUSION: This study demonstrated that C. pinnatifida presents important anti-inflammatory properties by inhibiting activated leukocytes and protein concentration of the exudate. These effects were related to the inhibition of proinflammatory mediators. The dicaffeoylquinic acids may be partially responsible for these anti-inflammatory properties through the inhibition of nuclear transcription factor kappa B and mitogen-activated protein kinase pathways.

"Arnicas" from Brazil: comparative analysis among ten species

Abstract The "arnicas" found in Brazil are examples of different species of the family Asteraceae used in popular medicine for its attributed anti-inflammatory action. Among the species known and used as "arnica" we selected: Calea uniflora Less., Chaptalia nutans (L.) Polák, Lychnophora ericoides Mart., Lychnophora pinaster Mart., Lychnophora salicifolia Mart., Lychnophora diamantinana Coile & S.B.Jones, Porophyllum ruderale (Jacq.) Cass., Pseudobrickellia brasiliensis (Spreng.) R.M.King & H.Rob., Sphagneticola trilobata (L.) Pruski, and Solidago chilensis Meyen, due to their extensive use. This research provides new information on leaf morphology and anatomy and on chemistry of the major metabolites found in these species through histochemical tests and phytochemical review. The results revealed anatomical characters for the differentiation and quality control of the vegetal drugs, being these: distinctive epidermal attachments, epidermis cells, parenchymal cells of the mesophyll, vascular bundles, midvein patterns and secretory structures of exudation of secondary metabolites. The review of chemical profiles showed differences in the chemical composition of the species, as different skeletons of sesquiterpene lactones in the species evaluated in addition to other chemical classes such as terpenes, flavonoids, chromenes and phenolic acids derivate. Based on the results obtained in this work it is important to emphasize that the information about the ten species of arnica generate subsidies for differentiation and identification of characteristic markers and for the diagnosis of the species and it can be applied in the "arnicas" quality control.

A single-step isolation by centrifugal partition chromatography of the potential anti-inflammatory glaucolide B from Lepidaploa chamissonis.

Sesquiterpene lactones (SL) are commonly found in Asteraceae and present a promising anti-inflammatory activity. Previously described in Lepidaploa genus, glaucolide B has never been investigated for its anti-inflammatory potential. This study aimed to establish an efficient process for the extraction of glaucolide B (1) from Lepidaploa chamissonis leaves and to develop a simple and fast method for its purification by using centrifugal partition chromatography (CPC), as well as to investigate in vitro the anti-inflammatory effects of glaucolide B. Thus, an optimized washing extractive process performed on L. chamissonis leaves allowed to obtain a SL enriched extract (4.11 g). After a successful defatting pretreatment of the crude extract, the glaucolide B enriched ethyl acetate portion (2.00 g) was fractionated by CPC affording, in a single-step isolation, compound 1 (1.04 g) in great yield (25%) and purity (97%). Cytotoxicity effect of 1 on RAW 264.7 macrophages was determined by using MTT assay, revealing a CC10 of 14.11 µM. Compound 1 at 1, 3 and 10 µM inhibited the nitrite/nitrate (NOx) metabolites production and the pro-inflammatory interleukin 6 (IL-6) secretion on lipopolysaccharide-stimulated RAW 264.7 cells. The extractive process used turned to be selective for SL and CPC technique proved a simple and effective tool for the isolation of 1 within few hours. Isolated for the first time from L. chamissonis leaves, glaucolide B presented a significant inhibitory effect on both NO and IL-6 secretion under non-toxic concentrations.

Calea uniflora Less. attenuates the inflammatory response to carrageenan-induced pleurisy in mice.

Calea uniflora Less. (family Asteraceae), also named "arnica" and "erva-de-lagarto", is a native plant to the South and Southeast of Brazil. This species was used to treat rheumatism, respiratory diseases, and digestive problems in Brazilian folk medicine. In vitro studies have shown the important biological effects of C. uniflora. However no studies have focused on the mechanism of action of anti-inflammatory activity of C. uniflora. The aim of this study was to evaluate the anti-inflammatory effects of the crude extract, its fractions, and isolated compounds obtained from of C. uniflora, using mouse model of carrageenan-induced inflammation. The following inflammatory parameters: leukocyte influx, degree of exudation, myeloperoxidase (MPO) and adenosine deaminase (ADA) activities, nitric oxide metabolites (NOx), proinflammatory cytokines and phosphorylation of the p65 subunit of NF-κB (p-p65 NF-κB), and p38 mitogen-activated protein kinase (p-p38 MAPK) levels were determined. The crude extract of C. uniflora, its fractions and its isolated compounds reduced the leukocyte influx, degree of exudation, MPO and ADA activities, NOx, TNF-α, IFN-γ, MCP-1 and IL-6 levels (p<0.05). The isolated compounds reduced p-p65 NF-κB and p-p38 MAPK levels (p<0.01). This study demonstrated that C. uniflora exhibits a significant anti-inflammatory activity via inhibition of the leukocyte influx and degree of exudation. These effects were associated with a decrease in the levels of several proinflammatory mediators. The mechanism of the anti-inflammatory action of C. uniflora may be, at least in part, via the inhibition of p65 NF-κB and p38 MAPK activation by the isolated compounds.

Beneficial effect of Ageratum conyzoides Linn (Asteraceae) upon inflammatory response induced by carrageenan into the mice pleural cavity.

ETHNOPHARMACOLOGICAL RELEVANCE: Ageratum conyzoides Linn (Asteraceae), a tropical plant that is very common in West Africa and some parts of Asia and South America, has been used to treat inflammatory disorders. In Brazil, teas made from A. conyzoides L. are used as anti-inflammatory, analgesic and anti-diarrheic agents. Therefore, it is necessary to study the mechanism of anti-inflammatory action of A. conyzoides L. to support its medicinal use for treating inflammatory conditions. These studies will also support the development of effective pharmacological agents with potent anti-inflammatory properties. AIM OF THE STUDY: To evaluate the anti-inflammatory effects of the crude extract (CE), its derived fractions: ethanol (EtOH-F), hexane (HEX-F), ethyl acetate (EtOAc-F) and dichloromethane (DCM-F) and isolated compounds, such as 5'-methoxy nobiletin (MeONOB), 1,2-benzopyrone and eupalestin, which are obtained from the aerial parts of A. conyzoides L. MATERIALS AND METHODS: These evaluations were performed using an animal model of inflammation induced by carrageenan. The following inflammatory parameters were analysed: leukocyte influx, protein concentration of the exudate, myeloperoxidase (MPO), adenosine deaminase (ADA) and nitric oxide metabolites (NOx) concentrations, interleukin 10 (IL-10), interleukin 17A (IL-17A), interleukin 6 (IL-6), tumor necrosis factor (TNF), interferon gamma (IFN-γ) and phosphorylation of p65 subunit of NF-κB (p-p65 NF-κB), p38 mitogen-activated protein kinases (p-p38 MAPK) were also analysed. RESULTS: CE, its EtOH-F, HEX-F, EtOAc-F and DCM-F and the isolated compounds, including MeONOB, 1,2-benzopyrone and eupalestin, significantly reduced leukocyte influx, protein concentration of the exudate, MPO, ADA, and NOx concentrations (p<0.05). CE, EtOH-F and isolated compounds significantly reduced IL-17A, IL-6, TNF and IFN-γ levels (p<0.05). CE, EtOH-F and isolated compound 1,2-benzopyrone also increased IL-10 levels (p<0.05). Isolated compounds, MeONOB, 1,2-benzopyrone and eupalestin, reduced p-p65 NF-κB and p-p38 MAPK (p<0.01). CONCLUSIONS: This study demonstrates that A. conyzoides L. exerts its important anti-inflammatory properties by inhibiting leukocyte influx and protein concentration of the exudate, as well as reducing the levels of several pro-inflammatory mediators. The anti-inflammatory action of A. conyzoides L. may be because of the inhibition of p65 NF-κB and MAPK activation by the isolated compounds.

Three new trixane glycosides obtained from the leaves of Jungia sellowii Less. using centrifugal partition chromatography.

Jungia sellowii (Asteraceae) is a shrub that grows in Southern Brazil and polar extract of its leaves presents anti-inflammatory properties. Cyperane, guaiane, nortrixane, and trixane sesquiterpene types were reported as the main metabolites in Jungia species. This work aims to describe the isolation and identification of sesquiterpenes in the leaves of J. sellowii using liquid-liquid partition and centrifugal partition chromatography. Thus, the crude extract of fresh leaves of J. sellowii was partitioned with hexane, dichloromethane, ethyl acetate and butanol, respectively. The butanol fraction was then subjected to a selected ternary system optimized for the CPC (centrifugal partition chromatography): ethyl acetate-ethanol-water (9:2:10, v/v/v). The separation was carried out isocratically at a flow rate of 25 mL/min at 1200 rpm, affording seven fractions A to G. TLC of fractions B, C and F displayed a single spot corresponding to three new glycosylated sesquiterpenoids. Their structures were established by using spectroscopic data in comparison to those reported in the literature. Furthermore, the isolates were evaluated for their leishmanicidal and cytotoxic effects. No cytotoxic effect was observed against the three cancer cell lines (HL60, JURKAT and REH), but compound 1 showed a weak antiprotozoal activity. Liquid-liquid partition and CPC turned to be a versatile technique of glycoside purification which is environmentally friendly and requires a limited amount of organic solvents.

Phytochemical analysis of Vernonanthura tweedieana and a validated UPLC-PDA method for the quantification of eriodictyol

AbstractVernonanthura tweedieana (Baker) H. Rob., Asteraceae, is used in the Brazilian folk medicine for the treatment of respiratory diseases. In this work the phytochemical investigation of its ethanol extracts as well as the development and validation of an UPLC-PDA method for the quantification of the eriodictyol from the leaves were performed. The phytochemical study for this species lead to the identification of ethyl caffeate, naringenin and chrysoeriol in mixture, eriodictyol from leaves, and the mixture of 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-propan-1-one and evofolin B, apigenin, the mixture of caffeic and protocatechuic acid and luteolin from stems with roots, being reported for the first time for V. tweedieana, except for eriodictyol. The structural elucidation of all isolated compounds was achieved by 1H and 2D NMR spectroscopy, and in comparison with published data. An UPLC-PDA method for quantification of the eriodictyol in leaves of V. tweedieana was developed and validated for specificity, linearity, precision (repeatability and intermediate precision), limit of detection (LOD) and limit of quantification (LOQ), accuracy and robustness. In this study, an excellent linearity was obtained (r2 = 0.9999), good precision (repeatability RSD = 2% and intermediate precision RSD = 8%) and accuracy (average recovery from 98.6% to 99.7%). The content of eriodictyol in the extract of leaves of V. tweedieana was 41.40 ± 0.13 mg/g. Thus, this study allowed the optimization of a simple, fast and validated UPLC-PDA method which can be used to support the quality assessment of this herbal material.

Jungia sellowii suppresses the carrageenan-induced inflammatory response in the mouse model of pleurisy.

This study was conducted to explore the anti-inflammatory effect of Jungia sellowii (Asteraceae) using a murine model of pleurisy induced by carrageenan (Cg). This plant is used in southern Brazil to treat inflammatory diseases. J. sellowii leaves were extracted with ethanol/water to obtain the crude extract (CE), which was fractionated with different solvents, yielding n-hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc) and n-butanol (BuOH) fractions, and aqueous fraction (Aq). The major compounds succinic acid (SA) and lactic acid (LA) were isolated from Aq fraction, and their structures were determined by (1)H and (13)C NMR. Pleurisy was induced by Cg (Saleh et al. 1996). The leukocytes, exudation, myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities, metabolites of nitric oxide (NO x ) levels, protein levels and mRNA expression for interleukin 1 beta (IL-1ß), tumour necrosis factor alpha (TNF-α), interleukin 17A (IL17A) and inducible of nitric oxide synthase (iNOs), and p65 protein phosphorylation (NF-κB) were analysed 4 h after pleurisy induction. Animals pre-treated with CE, BuOH, Aq, SA, or LA inhibited leukocytes, exudation, MPO and ADA activities, NO x , IL-1ß, TNF-α, and IL-17A levels, and the mRNA expression for IL-1ß, TNF-α, IL-17A, iNOS, and p65 protein phosphorylation (NF-κB) (p < 0.05). Our study demonstrated that J. sellowii can protect against inflammation induced by Cg by decreasing the leukocytes and exudation. Its effects are related to the decrease of either proinflammatory cytokines and/or NO x . The isolated compounds SA and LA may play an important role in this anti-inflammatory action by inhibiting all the studied parameters. The anti-inflammatory properties of these compounds are due to the downregulation of NF-κB.

Herbalists, traditional healers and pharmacists: a view of the tuberculosis in Ghana

This paper is the result of a visit by Brazilian researchers to Ghana, with the aim of improving understanding of the relationship between traditional healers and conventional health practices, specifically in relation to tuberculosis. Through this exploratory visit, this group of researchers promoted by the Edital Pro Africa (CNPq) had an opportunity to learn about, reflect on, and discuss the different social, economic and cultural realities and contexts that have led to the different health conditions and forms of healthcare in Ghana. Besides the direct relationship between the social and economic conditions of the country and the health of its population, it was also concluded that there is a clear distancing, in the Ghanaian reality, between the traditional healers and the conventional system, in terms of culture and modes of operation, each constituting isolated systems with little or no collaboration between them.The visit enabled us to see the difficulties involved in managing TB, including diagnosis, treatment, monitoring and co-infection with HIV. The majority of patients with TB only go to hospital after several attempts at self-medication, due to the non-specificity of the principal symptoms, and also to the trust in the traditional medicine. Initiatives to encourage research into medicinal plants in Ghana are seeking partnerships with developed countries, but not always with clear or secure national interests. For the traditional healers, there are high hopes that the information gathered by researchers from the local universities, on the plants and traditional methods they use, will result in affirmation and recognition of their practices, but they complain strongly that they receive no feedback on the research carried out.

Effects of the topical application of an ethyl acetate fraction from Vernonia scorpioides on excisional wounds infected with Staphylococcus aureus in rats

We investigated the healing process on excisional wounds infected with Staphylococcus aureus in rats, treated with 50 µL of ethyl acetate III from Vernonia scorpioides (Lam.) Pers., Asteraceae, rifamycin diethylamide B 25 mg, or saline. The lesions were measured daily and after seven days were surgically removed and histologically processed. The results indicate a favorable action of the EAIII, demonstrated by the increased wound contraction, smaller area of necrotic tissue, good development of granulation tissue, extensive extracellular matrix deposition and epithelial regeneration. This sub-fraction was phytochemically investigated in parallel studies, revealing the presence of sesquiterpene lactones (glaucolides and hirsutinolides) such as diacethylpiptocarphol and related hirsutinolides, flavonoids and cinnamic acid derivatives and also a new polyacetylene, which have been previously published. Results support the effectiveness of V. scorpioides antimicrobial activity in infected wound healing in rats.

Search for alkaloids on callus culture of Passiflora alata

Preliminary work on Passiflora alata leaves failed to detect harmane alkaloids using LC. The aim of this work was to investigate the production of harmane alkaloids through the cell culture of P. alata, inducing its precursor (L-tryptophan). The leaf explants presented satisfactory results after disinfection, and the callus formation was initiated in MS media with adequate quantities of phytohormones. Sixty days after inoculation, calli were inoculated in the optimized semi-solid MS media, with and without the addition of L-tryptophan (50, 100, 200 mg/L) and kept in standard conditions for 90 days. Calli were collected on days 6, 16, 26, 36, and 90, followed by acid-base extraction, and analysed by LC. The results showed an absence of harmane, harmin, harmol, harmalol, and harmaline. With L-tryptophan feeding, two peaks were detected, collected and analysed through positive mode electrospray [ESI(+)-MS] and sequential analysis in tandem ESI(+)-MS/MS. The spectra obtained were very similar, with a repetition of the more intense ions, and consecutive loss of 68 Da units, attributed to the heterocycle pyrazole. It appeared that this transformation was not related to any enzymatic pathway previously described for the plant from L-tryptophan, and the biosynthesis of β-carboline alkaloids in callus culture of P. alata were not observed in this work.

As folhas de varias espécies de Passiflora são utilizadas como ansioliticas e sedativas. Passiflora alata Curtis, Passifloraceae consta em três edições da farmacopéia brasileira, porem não há muitos estudos sobre sua composição química. No passado, enfatizava-se a ação conjunta de alcalóides e flavonóides. Em trabalho anterior, não foi detectada a presença de alcalóides harmanicos através de CLAE. Assim, decidiu-se investigar a produção dos mesmos através de cultivo celular, introduzindo seu precursor metabólico L-triptofano. Os explantes foliares apresentaram resultados satisfatorios para germinação apos assepsia, e a formação de calo foi iniciada em meio MS com quantidades adequadas de fitohormonios, previamente determinadas. Sessenta dias após a inoculação os calos foram repicados para meio semi-solido com e sem L-triptofano (50, 100, 200 mg/L), mantidos por 90 dias em condições padrão. Amostras foram coletadas com 6, 16, 26, 36, e 90 dias, realizada extração acido-base e o extrato analisado por CLAE. Os resultados mostraram a ausência de harmana, harmina, harmol, harmalol e harmalina. Dois picos presentes nas amostras com L-triptofano foram coletados e analisados através de espectrometria de massas, electrospray modo positiva [ESI(+)-MS] e analise em tandem ESI(+)-MS/MS. Os espectros correspondentes foram similares, mostrando a perda consecutiva de 68 Da, atribuídos ao pirazol. Este fato aponta para uma transformação não enzimática, não relacionada a uma biossintese previamente descrita para alcalóides β-carbolínicos.

Evaluation of the proliferative activity of methanol extracts from six medicinal plants in murine spleen cells

A number of natural compounds have been used as immunomodulatory agents, enabling the function of the immune system to be modified by stimulating or suppressing it. There has been increasing interest in the study of therapeutic action of plant extracts regarding their immunomodulatory activity. The aim of this study was to identify and evaluate the action of extracts of the medicinal plants Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis and Vernonia scorpioides on the development of spleen cells from mice, using the in vitro cellular proliferation assay. The cells, obtained by mechanical rupture of mice spleen (5x10(4) cells/mL), were incubated with methanol extracts (10, 50, 100 and 200 µg/mL) and phytohemagglutinin (PHA, 5 µg/mL). The basal control for proliferation consisted of cells alone, while the positive control consisted of cells and PHA. The cell culture was kept at 37 ºC in 5 percent CO2 for 72 hours, and cell proliferation was revealed by the blue tetrazolium reduction assay (MTT). The results were expressed as percentage of growth and were analyzed using the Kruskal-Wallis and Mann-Whitney tests. The C. brasiliense, I. pes-caprae and M. elaeagnoides extracts showed dose-dependent induction of cell proliferation, with a significant increase in cell proliferation (p<0.03) and percentage growth of 88.2 percent, 73.1 percent and 52.7 percent, respectively, suggesting T lymphocyte stimulation. By contrast, M. robusta, R. imperialis and V. scorpioides extracts showed significance only with a negative percentage of growth, suggesting inhibition of cell proliferation (p<0.04). Further biomonitoring studies will enable the fractions and isolated substances responsible for the immunomodulatory activities to be identified.

Várias substâncias de origem natural têm sido utilizadas como agentes imunomoduladores, permitindo modificar a função do sistema imune e propiciando o estudo de atividades terapêuticas de extratos de plantas. Este trabalho objetivou identificar a atividade imunomodulatória dos extratos de seis plantas medicinais da flora brasileira, Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis e Vernonia scorpioides, sobre a proliferação de células esplênicas de camundongos. As células esplênicas murinas obtidas por ruptura mecânica do baço (5x14³ células/mL) foram incubadas com os extratos metanólicos das plantas (10, 50, 100, 200 µg/mL) e fito-hemaglutinina (PHA, 5 µg/mL). O controle basal de proliferação foi constituído de células apenas e o controle positivo formado por células e PHA. O cultivo celular foi mantido a 37 ºC, 5 por cento de CO2, 72 horas, com quantificação da proliferação celular pelo ensaio de redução do azul de tetrazólio. Os resultados expressos em percentagem de crescimento foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney. Os extratos de C. brasiliense, I. pes-caprae e M. elaeagnoides mostraram indução dose-dependente da proliferação celular (p<0,03), com percentagem de crescimento de, respectivamente, 88,2 por cento, 73,1 por cento e 52,7 por cento, sugerindo estímulo de linfócitos T. Contrariamente, os extratos de M. robusta, R. imperialis e V. scorpioides apresentaram significância apenas com percentagem de crescimento negativa, indicando inibição da proliferação celular (p<0,04). A continuidade no estudo biomonitorado permitirá a identificação das frações e substâncias isoladas responsáveis pelas atividades imunomoduladoras.

Anti-inflammatory evaluation of Coronopus didymus in the pleurisy and paw oedema models in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Mastrunço (Coronopus didymus--CD) is currently considered as a medicinal specie often used in Brazil, especially in southeast region, for the treatment of several diseases in which pain and inflammation are common. Treatment with the plant can be done by infusion, decoction, or through food. The aim of this study was: to investigate the anti-inflammatory effect of hydroalcoholic extract obtained from the leaves of CD following the traditional procedure. MATERIALS AND METHODS: The anti-inflammatory activity was determined using mouse of pleurisy and paw oedema models, both process being induced by different flogistic agents such as: carrageenan (Cg), bradykinin (BK), histamine (HIS), substance P (SP), dextran (DEX) or prostaglandin E(2) (PGE(2)). We evaluated the effect of CD (200-600 mg/kg) administered by oral route (p.o.) upon leukocytes migration, myeloperoxidase (MPO), and adenosine-deaminase (ADA) activities and nitric oxide (NO) levels. RESULTS: CD (200-600 mg/kg) inhibited the leukocytes by 60.0+/-1.42%, neutrophils by 82.75+/-1.29%, MPO by 42.30+/-4.23%, and ADA activities by 57.89+/-1.94%, as well as NO levels by 64.28+/-2.15% in Cg induced pleurisy. CD also inhibited total and differential leukocytes in the pleurisy induced by BK (1.30+/-0.11/0.29+/-0.02), HIS (1.20+/-0.09/0.42+/-0.05) and SP (0.74+/-0.06/0.14+/-0.01). In addition, CD was effective in reducing paw oedema induced by Cg by 72.79+/-1.13%, SP by 68.26.+/-0.78%, BK by 66.66.+/-0.77%, PGE(2) by 53.346.+/-1.18 and DEX by 65.14+/-2.35%. CONCLUSION: Several mechanisms, including the inhibition of enzymes (MPO and ADA) and mediators (BK, HIS, SP, NO and PGE(2)) release and/or action, appear to account for the anti-inflammatory effect of Coronopus didymus.