Effects of a nanoemulsion with Copaifera officinalis oleoresin against monogenean parasites of Colossoma macropomum: A Neotropical Serrasalmidae.

Monogeneans are ectoparasites that may cause losses in production and productivity in the aquaculture of Colossoma macropomum. Chemotherapeutics used in aquaculture usually have major adverse effects on fish; hence, the use of essential oils has been considered advantageous, but these are not soluble in water. Thus, the use of nanostructures to enhance water solubility of compounds and improve bioactivity may be very promising. This study investigated the antiparasitic activity of nanoemulsion prepared with Copaifera officinalis oleoresin (50, 100, 150, 200 and 300 mg/L), against monogenean parasites from the gills of C. macropomum. The particle size distribution and zeta potential suggested that a potentially kinetic stable system was generated. The nanoemulsion from C. officinalis oleoresin achieved high efficacy (100%) at low concentrations (200 and 300 mg/L) after 15 min of exposure. This was the first time that a nanoemulsion was generated from C. officinalis oleoresin using a solvent-free, non-heating and low-energy method. Moreover, this was the first time that an antiparasitic against monogeneans on fish gills, based on nanoemulsion of C. officinalis oleoresin, was tested.

First cytotoxic, genotoxic, and antigenotoxic assessment of Euterpe oleracea fruit oil (açaí) in cultured human cells.

Euterpe oleracea Mart., popularly known as "açaí", is a tropical fruit from the Amazon region where it has considerable economic importance. Açaí has been used as food and for several medicinal purposes. Despite the widespread use of this fruit, there is a lack of data regarding the safety of using this fruit oil exclusively. Therefore, we evaluated the in vitro cytotoxic, genotoxic, and antigenotoxic effects of E. oleracea fruit oil (EOO) in cultured human lymphocytes (non-metabolizing cells) and HepG2 cell line (human hepatoma) (metabolizing cells) by using MTT, comet, and micronucleus assays. A wide range of EOO concentrations was tested with a preliminary MTT assay, which allowed selecting five concentrations for comet and micronucleus assays: 2.5, 10, 100, 500, and 1000 µg/mL. The results showed that none of the EOO tested concentrations presented cytotoxic effects. The genotoxic assessment revealed an absence of significant DNA and chromosome damage in human lymphocytes and HepG2 cells but did not show chemoprotection against the DNA damage induced by methyl methanesulfonate and benzo[a]pyrene, used as DNA-damaging agents.

In vitro study of the cytotoxic and genotoxic effects of indomethacin-loaded Eudragit(®) L 100 nanocapsules.

Indomethacin is a non-steroidal anti-inflammatory agent included in one of the most commonly used drug classes worldwide. The use of this drug results in certain side effects, including gastrointestinal complications. Therefore, there exists a need to develop better methods for the delivery of such drugs into the body, such as those employing nanoparticles. The aim of the present study was to evaluate the cytotoxic and genotoxic effects of indomethacin-loaded Eudragit(®) L 100 nanocapsules (NI; based on methacrylic acid and methyl methacrylate) on cells unable (lymphocytes) and able to metabolize drugs (HepG2 cells), using comet and cytokinesis-block micronucleus (CBMN) assays in vitro. Cells were exposed to NI at concentrations of 5, 10, 50, 125, 250, and 500 µg/mL. The comet assay showed that NI induced no significant DNA damage in either cell type at any of the concentrations tested. The CBMN test confirmed these results; however, the highest concentration of 500 µg/mL resulted in a small but statistically significant clastogenic/aneugenic effect in HepG2 cells. These findings should encourage the development of new investigations of this nanomaterial as a delivery vehicle for anti-inflammatory drugs, such as indomethacin.

Evaluation of the genotoxicity of Euterpe oleraceae Mart. (Arecaceae) fruit oil (açaí), in mammalian cells in vivo.

E. oleracea is a tropical plant from the Amazon region, with its fruit used for food, and traditionally, as an antioxidant, anti-inflammatory, hypocholesterolemic, for atherosclerotic disease, and has anticancer properties. The oil of the fruit has antidiarrheic, anti-inflammatory and antinociceptive activities, but without genotoxicity evaluation. Therefore, the aim of this study was to evaluate the genotoxic potential of E. oleracea fruit oil (EOO), in rat cells. Male Wistar rats were treated with EOO by gavage at doses of 30, 100 and 300 mg/kg, for 14 days, within a 24 h interval. The DNA damage in the leukocytes, liver, bone marrow and testicular cells, was assessed by the comet assay, and the clastogenic/aneugenic effects in the bone marrow cells, by the micronucleus test. Our phytochemicals characterization of the EOO showed the presence of vanillic, palmitic, γ-linolenic, linoleic, oleic, cinnamic, caffeic, protocatechuic, ferulic, syringic acids, and flavonoids quercetin and kaempferol rutinoside as the main constituents. Both cytogenetic tests performed showed that EOO presented no significant genotoxic effects in the analyzed cells, at the three tested doses. These results indicate that, under our experimental conditions, E. oleracea fruit oil did not reveal genetic toxicity in rat cells.

Pre-clinical validation of a vaginal cream containing copaiba oil (reproductive toxicology study).

The aims of this study was to evaluate the effects of oil-resin of Copaiba (Copaifera duckei Dwyer), aired in vaginal cream on the reproductive performance of female rats (Rattus norvegicus). To determine the components of the C. duckei oleoresin, gas chromatography coupled with mass spectrometry (CG-MS) was used, and considering the trans-caryophyllene sesquiterpene as a phytochemical marker in the oleoresin. Due to the extensive use of copaiba oleoresin in the suppository form for gynecological infections, an evaluation was carried out on the effects of copaiba oleoresin (Copaifera duckei Dwyer), delivered in a vaginal cream, on the reproductive performance of female Wistar rats. For this purpose, three groups (n=5-6/group) of female rats were treated as follows: 1--vaginal cream of copaiba oleoresin (28.6 mg/kg), 2--base vaginal cream and 3--control (physiological saline 0.9%), administered intravaginally, for 30 days before pregnancy, and from day zero to day 20 during pregnancy. Laparotomy was performed on the 21st day of pregnancy, followed by the determination of reproductive variables: number of live and dead fetuses, mass of the fetuses and placentas, number of implantations and resorptions, number of corpora lutea, pre- and post-implantation loss, and analyses of the fetuses with regard to external and internal anomalies and/or malformations (skeletal and visceral). The trans-caryophyllene present in the sample is suggested as a phytochemical marker and the results of this study demonstrate an absence of maternal toxicity and foetotoxicity embryofoetotoxicity at the dose administered, corresponding to ten times the recommended dose for use in humans. Accordingly, no significant statistical difference was observed between the treated and control groups, for the variables analyzed. Thus, it is concluded that the vaginal cream containing 2.5% copaiba oleoresin is safe during gestation, in female rats (Rattus norvegicus) of the Wistar strain.

Anti-inflammatory and cicatrizing activities of a carbohydrate fraction isolated from sugary kefir.

Kefir is an association of microrganisms generally grown in milk, with known probiotic activities identified from its soured suspensions. Aqueous media are also able to grow kefir, but little is known about the probiotic properties of its fermented products. This work aimed to evaluate some probiotic properties of a carbohydrate fraction isolated from sugary kefir (sugary kefir carbohydrate [SKC]). Anti-inflammatory activity of the isolated fraction of carbohydrate was tested both in vitro (cellular respirometry and macrophage culture) and in vivo (50% effective dose, rat paw edema, vascular permeability, and cicatrizing test). The results indicated no significant difference for oxygen uptake or macrophage culture between control and test groups. Rat paw edema, however, showed a significant inhibitory activity by 30 +/- 4% and 54 +/- 8% (P < .001) for carrageenan and dextran, respectively. In the cicatrizing test, animals treated with SKC cream also presented less trauma after treatments as compared to the negative control group (P < .05). The overall data suggested the SKC as a natural product that could be used as a constituent of an anti-inflammatory compound.

Amazonian plant crude extract screening for activity against multidrug-resistant bacteria.

Antimicrobial resistance is a subject of great concern in public health and also in the designing of strategies for current therapeutic protocols all over the world. New drugs, including those necessary for a reserve armamentarium and exhibiting less side effects deserve special attention. In rural areas, particularly in Brazil, a huge number of natural products, in different artisanal preparations, mainly from plants, have been used by traditional populations to cure diseases. Despite some of these plants have been studied, many of them are awaiting to have their compounds chemically characterized and investigated their pharmacodynamics properties. Further, as well known, the environment plays a crucial role in the metabolism of these plants, yielding different and varied molecular complexes depending on the period of collection, climate conditions, kind of soil and also the plant speciation. In this report, ethanol crude extract of 10 different botanical specimens from the Amazon region of Brazil, in the Amapa State, were screened for antibacterial activity of 7 clinical resistant microorganisms utilizing as control ATCC bacterial species by the Kirby-Bauer method. Plant extracts of Geissospermum argenteum, Uncaria guianensis, Brosimum acutifolium, Copaifera reticulate, Licania macrophylla, Ptycopetalum olacoides and Dalbergia subcymosa yielded activity against Staphylococcus aureus and Pseudomonas aeruginosa, both multidrug resistant, and Staphylococcus aureus ATCC strain.

In vivo study of the anti-inflammatory effect of Rhus toxicodendron.

BACKGROUND: Homeopathic Rhus toxicodendron (Rhus tox) is used in various inflammatory conditions. We screened its effect compared to succussed ethanol controls and appropriate active controls. METHOD: We initially experimented with Rhus tox 6, 12, 30 and 200 cH, using carrageenan-induced paw oedema in rats. The 6 cH dilution appeared most effective and was used in subsequent assays. We used pre-treatment and single treatment regimes in Wistar rats, and mice. RESULTS: We found significant reductions compared to control in carrageenan-induced paw oedema, vascular permeability, writhing induced by intraperitoneal acetic acid and stress induced gastric lesions. CONCLUSIONS: Rhus tox in homeopathic dilution appears to interfere with inflammatory processes involving histamine, prostaglandins and other inflammatory mediators.

Anti-inflammatory and antinociceptive activities of extract, fractions and populnoic acid from bark wood of Austroplenckia populnea.

Austroplenckia populnea (Reiss) Lund is a Brazilian plant from "cerrado", which belongs to Celastraceae family, popularity know as "marmelinho-do campo, mangabeira-brava, mangabarana, vime and maria-mole". This plant is used in folk medicine to treat dysenteries and inflammatory disorders, such as rheumatism. Austroplenckia populnea bark hydroalcoholic crude extract, and its hexane, chloroform and ethyl acetate fractions, obtained by partition, as well as the isolated populnoic acid were investigated for their anti-inflammatory (carrageenan, dextran and histamine-induced rat paw oedema, histamine-induced increase in vascular permeability, and granulomatous tissue induction) and analgesic activities (writhing and hot plate tests). The ED(50) (oral) of the crude extract for the inhibition of carragenan-induced rat paw oedema assay was determined to be 200 mg/kg, which was also used in the assays with the extract and its fractions in all other experiments. Populnoic acid was administered in the dose of 50 mg/kg. Crude extract, hexane and chloroform fractions (200 mg/kg), and indomethacin (10 mg/kg) inhibited significantly (p<0.05) the formation of the carrageenan-induced rat paw oedema, measured in third hour of experiment (peak of oedema formation) by 43.2%, 37.3%, 31.1% and 59.3%, respectively. There was a significant reduction (p<0.05) in dextran-induced rat paw oedema in all groups, while in the assay using histamine as the oedematogenic agent, only the groups treated with populnoic acid (50 mg/kg) and cyproheptadine (10 mg/kg) displayed significant reduction (p<0.05). The populnoic acid and cyproheptadine reduced the peak of oedema formation (1st hour) by 41.3% and 34.7%, respectively. Only for the groups treated with populnoic acid (50 mg/kg) and cyproheptadine (10 mg/kg) it was observed a significant (p<0.05) reduction in histamine-induced increase in vascular permeability (44.8% and 80.3%, respectively). Granulomatous tissue formation was significantly inhibited (p<0.05) by both hexane fraction (46.0%) and dexamethasone (66.2%). In the analgesic assays, the crude extract and its hexane and chloroform fractions, as well as indomethacin diminished significantly the number of writhings (p<0.05) by 69.6%, 47.2%, 44.8% and 62.8%, respectively. On the other hand, none assayed sample displayed significant result in the hot plate test. Based on the obtained results it is suggested that extracts of Austroplenckia populnea bark and populnoic acid display anti-inflammatory activity, supporting its folkloric use to treat inflammatory disorders.

Topical antiinflammatory and analgesic activities of Copaifera duckei dwyer.

The oleoresin of several Copaifera species is used widely in the Amazonian Region mainly as a topical antiinflammatory and healing agent. The topical analgesic and antiinflammatory activities of Copaifera duckei oleoresin, whose terpenoidal chemical composition has been characterized, are now examined. Antiinflammatory activity was evaluated in rats using the carrageenin-induced paw edema and the granuloma tests, and in mice by the croton oil-induced dermatitis test. Analgesic activity was determined in mice using the writhing test method. In the carrageenin-induced edema and granuloma tests the oleoresin in a dose of 1,802 mg/kg inhibited the edema by 18% and granuloma by 42% (p < 0.05), this last result similar to that observed with dexamethasone. Topical doses of 517 mg/kg, 1,035 mg/kg and 1,802 mg/kg produced 52%, 58% and 62% (p < 0.05) reduction of the edema induced by croton oil, respectively, and 48%, 56% and 65% inhibition of the writhing process (p < 0.05). These results suggest that the Copaifera duckei oleoresin has topical antiinflammatory and analgesic activities.

Analgesic and anti-inflammatory activity of a crude root extract of Pfaffia glomerata (Spreng) Pedersen.

The anti-inflammatory and antinociceptive effects of the crude hydroalcoholic extract (PE) of Pfaffia glomerata roots was assessed in the carrageenan-induced rat paw edema at the doses of 100, 200 and 300 mg/kg, using different animal models. An anti-inflammatory dose effect response correlation of r=0.997 and Y=11.67x+0.02 was found. At the same doses, the extract-inhibited acetic acid-induced writhing in mice, but no dose response correlation was found. Oral administration of 100 mg/kg of PE and 0.5 mg/kg of dexamethazone inhibited by 29 and 61%, the granulomatous tissue formation (p>0.05), respectively. These results indicate the potential of this plant extract to treat chronic inflammation. At the assayed doses no significant activity was found in the hot plate test, as well as in the cell migration-induced by carrageenan.

Effect of Atropa belladonna and Echinacea angustifolia in homeopathic dilution on experimental peritonitis.

Atropa belladonna and Echinacea angustifolia have been used in homeopathy as modulators of inflammatory processes, in simple potency or 'accord of potencies', as recommended by homotoxicology. We evaluated their effects on leukocyte migration and macrophage activity induced by experimental peritonitis in vivo. Mice were injected (i.p.) with LPS (1.0mg/kg) and treated (0.3ml/10g/day, s.c.) with different commercial forms of these medicines. Echinacea angustifolia D4--a simple potency preparation--and Belladonna Homaccord, Belladonna Injeel, Belladonna Injeel Forte, Echinacea Injeel and Echinacea Injeel Forte--all in 'accord of potencies'--were tested. The association of A. belladonna and E. angustifolia in 'accord of potencies' produced an increase of polymorphonuclear cell migration (Kruskal-Wallis, P = 0.03) and a decrease of mononuclear cell percentages (Kruskal-Wallis, P < or = 0.04), when compared with control, mainly in preparations containing low potencies. The proportion of degenerate leukocytes was lower in the treated groups, compared to a control group (P < or = 0.05). The treated groups showed increased phagocytosis (P < or = 0.05), mainly in preparations containing high potencies. Our results suggest that A. belladonna and E. angustifolia, when prepared in 'accord of potencies', modulate peritoneal inflammatory reaction and have a cytoprotective action on leukocytes.

Evaluation of the genotoxic potential of the Casearia sylvestris extract on HTC and V79 cells by the comet assay.

Casearia sylvestris is common in tropical America growing wild in Brazil in the states of Amazonas and São Paulo. Its leaves are used in Brazilian folk medicine for several diseases. The present investigation was carried out to examine the genotoxic effects of a C. sylvestris crude ethanolic extract on Hepatoma Tissue Culture (HTC cells) of Rattus norvegicus and Chinese hamster V79 cells in culture, using the comet assay. For the genotoxic evaluation the cells were treated with three different concentrations (0.5, 1 and 2 mg/ml) of extract prepared from a 25 mg/ml aqueous solution. The positive control was cyclophosphamide for HTC cells and methyl methanesulfonate for V79 cells. The duration of the treatment was 2 h. The results showed that the extract of C. sylvestris presented no genotoxic effects and not modified effect inducing DNA damage by alkylating agents cyclophosphamide and methyl methanesulfonate in HTC and V 79 cells respectively.

Analgesic and anti-inflammatory activities evaluation of (-)-O-acetyl, (-)-O-methyl, (-)-O-dimethylethylamine cubebin and their preparation from (-)-cubebin.

The anti-inflammatory and antinociceptive effects of the acetylated (2), methylated (3) and aminated (4) derivatives of cubebin (1), obtained by its reaction with acetic anhydride, methyl iodide and dimethylethylamine chloride, respectively, were investigated, using different animal models. The compound (2) was the most effective anti-inflammatory one in the carrageenin-induced paw edema in rats and was the only one which showed dose-response correlation for this assay with r = 0.993 and Y = 64.58x + 0.22. Besides, compounds (2) and (4) were more effective than cubebin in inhibiting acetic acid-induced writhing in mice, producing dose-response correlation with doses of 10, 20 and 30 mg/kg, respectively. Regarding the hot plate and the cell migration tests in rats, none of the four tested compounds showed activity. Overall, the results showed that the acetylation and amination of cubebin were efficient in enhancing its analgesic activity, as well as its anti-inflammatory activity.

Central properties of the essential oil and the crude ethanol extract from aerial parts of Artemisia annua L.

The present study evaluated the central activity of the essential oil and the ethanolic extract from Artemisia annua L. in animals as a part of a psychopharmacological screening of this plant. The extract was prepared with fresh leaves in ethanol (AEE) and the essential oil (AEO) was obtained by hidrodestilation. The ED(50) and the LD(50) obtained for the essential oil were 470mg/kg (correlation coefficient r=0.97333 and linear regression y=-26.52x+0.158) and 790mg/kg, and for the extract, 450mg/kg (correlation coefficient r=0.99266 and linear regression y=-27.34+0.156) and more than 2g/kg, respectively. The doses increased the latency time to convulsions induced by picrotoxin and pilocarpine but prevented the onset of pentylenotetrazol and strychnine induced seizures. In addition to, the products have caused marked inhibition in the Rota-rod assay. According to the results, the AEO has a high acute toxicity and a possible cholinergic action, and the AEE showed a possible central activity as dopaminergic and cholinergic agents, and did not present a significant acute toxicity. These differences should be due to chemical substances present in each product. These products had no significant effect as an anticonvulsant, while exhibited a strong depressant activity on the CNS.

Study of anti-inflammatory activity of Tibetan mushroom, a symbiotic culture of bacteria and fungi encapsulated into a polysaccharide matrix.

Tibetan mushroom (TM) is a fermented beverage composed by a dozen of bacteria and yeasts living together into polysaccharide grains secreted by them. TM is similar to kefir, a probiotic beverage originated in the Caucasian mountains exhibiting some anti-bacterial, anti-mycotic, anti-neoplastic and immunomodulatory effects. Aiming to evaluate a plausible anti-inflammatory property of TM we conducted cotton-induced granuloma and paw edema assays in rats, the latter using carrageenin, dextran and histamine as stimuli. TM samples were thawed and continuously cultured during 15 days into molasses solutions (50 g/l). The experiments used TM suspensions after 24h fermentation and TM grains mechanically disintegrated. The results showed a significant inhibition on the formation of granuloma tissue for the test group as compared to the negative control group. TM suspensions presented an inhibition of 43% for the inflammatory process. Rat paw edema also showed significant decreases with the mediators. The edema induced by carrageenin was inhibited 62% at the 3rd hour. The edema dextran-induced was completely inhibited at 1h and antagonized the histamine edema 52% at 1h.

Atividade antiinflamatória do granulado de Calendula officinalis L. e Matricaria recutita L.

Este trabalho descreve os resultados da atividade antiinflamatória de um granulado composto pelos extratos secos de Calendula officinalis L. e Matricaria recutita L. (GECOMR® - granulado encapsulado de Calendula officinalis e Matricaria recutita). Estas espécies são utilizadas na medicina popular devido às suas atividades antiinflamatória, antiúlcera gástrica e antiespasmódica. A redução do processo edematogênico nos grupos de animais tratados com o GECOMR foi observada para todos agentes inflamatórios testados (Carragenina, dextrana e histamina). Esta associação mostrou-se eficaz sobre os modelos de edema de pata induzidos pelas diferentes substâncias inflamatórias, com as doses de 100 e 250 mg.kg-1. A associação dos extratos hidroalcoólicos destas espécies medicinais não modificou a indicação isolada de cada uma.

This study shows the results of the anti-inflammatory activity of a phytocomplex composed by dry extracts of Calendula officinalis L. and Matricaria recutita L. (GECOMR®). These plants have been used by folk medicine for its anti-inflammatory, antiulcerogenic and antispasmodic activities. The reduction of the edematogenic process in the treated animals with GECOMR was observed for all inflammatory agents tested, (Carrageenan, Dextran and Hystamine). Its association has been efficient in rat paw edema induced by inflammatory substances in 100 and 250 mg.kg-1 doses. This association of the ethanolic extracts did not modify the indication of each plant isolated.

Atividade antiinflamatória de quefir, um probiótico da medicina popular

O quefir é uma bebida fermentada originária do Cáucaso e formada por bactérias ácido-lácticas e leveduras, sendo utilizada por suas propriedades organolépticas e terapêuticas. O presente estudo avaliou a atividade antiinflamatória deste probiótico sobre modelo de indução de tecido granulomatoso e contorções abdominais induzidas por ácido acético, in vivo. Os resultados obtidos mostraram que o quefir inibiu a formação do tecido granulomatoso, bem como o número de contorções abdominais causadas por ácido acético, quando comparado ao grupo controle. Desta forma, pode-se sugerir uma possível ação antiinflamatória.

Kefir is a fermented beverage with its origin in Caucasian region, formed by acid-lactic bacteria's and yeasts, and used due to its organoleptic and therapeutic properties. The present study evaluated the possibility of an anti-inflammatory activity for induction of granulomatous tissue in rats and writhing test in mice. The results obtained shown that the kefir has inhibited the formation of the granulomatous tissue, as well as the number of writhings induced by acetic acid as compared to a control group. In this way the authors suggest a possible antiinflammatory action.