Nutritional and chemical composition of Alpinia zerumbet leaves, a traditional functional food.

Alpinia zerumbet, a species of the Zingiberaceae family, is a common plant in tropical and subtropical areas used in traditional medicine to treat various diseases and also included as food in the traditional Okinawan diet (Japan). The leaves and rhizomes of this plant are used as spice and flavoring in foods such as rice, meats, and pasta. Studies of the chemical and nutritional characteristics of fresh leaves and of leaves submitted to thermal treatments such as boiling and steaming are lacking. In the current study, the leaves of A. zerumbet were subjected to boiling or steaming for 10, 20, and 30 min as part of the thermal treatments. The study also provides noteworthy results regarding the proximate composition, physical-chemical data, minerals, phenolic compounds, antioxidant activity, volatile compounds, and LC-MS chromatographic profiles of the extracts produced with fresh leaves and with thermal treatments. The carbohydrate content of A. zerumbet leaves improved during the thermal treatments, showing an increase after steaming (18.86 to 19.79%) and boiling for 30 min (25.85%). After boiling treatment for 20 min, a significant amount of protein was found (6.79%) and all heat treatments resulted in low content of lipid (<1.0%). The boiling treatment for 10 min (BT10) resulted in the highest concentrations of total phenolic components (TPC), 339.5 mg/g, and flavonoids (TF), 54.6 mg/g, among the three thermal treatments (BT10, BT20 and BT30). The results of the steaming treatments (ST 10, 20, and 30 min) differed, with ST20 leading to higher TPC (150.4 mg/g) and TF (65.0 mg/g). The quantity of total phenolics and flavonoids, as well as the antioxidant activity, were significantly affected by the cooking method and the length of time of sample exposure to heat. The samples boiled for 30 and 10 min had higher concentrations of antioxidant activity as measured by the phosphomolybdenum and DPPH methods (151.5 mg/g of extract and 101.5 µg/mL, respectively). Thirty-eight volatile organic compounds (VOCs) were identified by chromatographic analysis of fresh and thermally treated leaves of A. zerumbet. Terpenoids were the predominant class of volatile compounds in the fresh leaves and in all thermal treatments. p-Cymene, 1,8-cineole, 4-terpineol, linalool, α-copaene and ß-bisabolene have the greatest impact on overall aroma perception, with odor activity values (OAV) greater than five. Among the phenolic compounds identified by LC-HRMS in the extracts of fresh and thermally treated leaves were proanthocyanidins, (+) catechin, (-) epicatechin, quercetin-3-O-glucoronide, isorhamnetin-3-O-glucoronide, kaempferol-3-O-rutinoside, pinocembrin, alpinetin, pinostrobin, and other compounds. The present results support the traditional use of this plant as a potential food with properties that certainly contribute to health improvement.

Evaluation of the Amazonian fruit Ambelania acida: Chemical and nutritional studies.

Ambelania acida is native to the Amazon region, with few published studies of its fruits. We examined the proximate composition of its fruits, including minerals, fatty acids, volatile organic compounds (VOCs), as well as its antioxidant capacity. The protein contents (2.61%) of the pulp and seeds (13.6%) were higher than observed in other taxa of the family or in other tropical fruits. Peel and pulp showed high contents of potassium, calcium, and magnesium, and the potassium content in the pulp was 1125 mg/100 g. The peel had higher contents of total phenolics, tannins, and ortho-diphenols than the pulp, as well as better antioxidant activity as evidenced by 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), and Fe2+ chelating activity assays. GC-MS analyses identified 42 VOCs in the peel and pulp, with more than 90% being classified as terpenes. Eleven types of fatty acids were identified in the lipid fractions of the peel, pulp, and seeds. Linoleic acid, an essential fatty acid for humans, was the principal fatty acid in the edible portion of the fruit, therefore, evidencing its nutritionally significant profile for the fruits when considering the relationship among polyunsaturated, saturated, and monounsaturated fatty acids. The information gathered here indicates that this native fruit is a healthy food source and its cultivation and consumption should be stimulated.

Endlicheria bracteolata (Meisn.) Essential Oil as a Weapon Against Leishmania amazonensis: In Vitro Assay.

The difficulties encountered and the numerous side effects present in the treatment of cutaneous leishmaniasis have encouraged the research for new compounds that can complement or replace existing treatment. The growing scientific interest in the study of plants, which are already used in folk remedies, has led our group to test Endlicheria bracteolata essential oil against Leishmania amazonensis. Several species of the Lauraceae family, or their compounds, have relevant antiprotozoal activities Therefore, the biological potential on L. amazonensis forms from the essential oil of Endlicheria bracteolata leaves was verified for the first time in that work. The antileishmanial activity was evaluated against promastigotes and intracellular amastigotes, and cytotoxicity were performed with J774.G8, which were incubated with different concentrations of E. bracteolata essential oil. Transmission electron microscopy and flow cytometry were performed with E. bracteolata essential oil IC50. Promastigote forms showed E. bracteolata essential oil IC50 of 7.945 ± 1.285 µg/mL (24 h) and 6.186 ± 1.226 µg/mL (48 h), while for intracellular amastigote forms it was 3.546 ± 1.184 µg/mL (24 h). The CC50 was 15.14 ± 0.090 µg/mL showing that E. bracteolata essential oil is less toxic to macrophages than to parasites. Transmission electron microscopy showed that E. bracteolata essential oil treatment is capable of inducing mitochondrial damage to promastigote and intracellular amastigote forms, while flow cytometry showed ΔÑ°m disruption in treated parasites. These results could bring about new possibilities to develop products based on E. bracteolata essential oil to treat cutaneous leishmaniasis, especially for people who cannot receive the conventional therapy.

Wound healing properties of Copaifera paupera in diabetic mice.

Copaifera oleoresin is one of the most used natural products in popular medicine all over the world. Among other effects (i.e., anti-inflammatory, antinociceptive, microbicidal) one of the most well-known is its wound healing capacity. However, the mechanism by which the oleoresin presents its effect is still not clear. In this study, our aim was to evaluate the wound healing capacity of oleoresin obtained from Copaifera paupera, its mechanism of action and identify its major components. For these purposes, diabetic Swiss Webster mice were topically treated with oleoresin (100, 150 or 200 mg/kg) for 14 consecutive days after an excision was performed in the back of the mice. Cytokines, wound retraction and histological evaluation were conducted at 3, 7 and 10 days (for cytokines); 0, 3, 7, 10 and 14 days (for wound retraction); and 7 and 14 days (for histological evaluation). Our data indicate that oleoresin significantly reduced production of MCP-1 and TNF-α at days 7 and 10 post-excision and increased IL-10 production at both days. All treatments demonstrated an effect similar or higher to that in collagenase-treated mice. Histological evaluations demonstrated that higher dose treatment resulted in better resolution and closure of the wound and higher levels of collagen deposition and indexes of re-epithelialization even when compared with the collagenase-treated group. The treatment with oleoresin from Copaifera paupera demonstrated that it is even better than an ointment routinely used for improvement of wound healing, suggesting this oleoresin as an option for use in diabetic patients.

Natural Products: Insights into Leishmaniasis Inflammatory Response.

Leishmaniasis is a vector-borne disease that affects several populations worldwide, against which there are no vaccines available and the chemotherapy is highly toxic. Depending on the species causing the infection, the disease is characterized by commitment of tissues, including the skin, mucous membranes, and internal organs. Despite the relevance of host inflammatory mediators on parasite burden control, Leishmania and host immune cells interaction may generate an exacerbated proinflammatory response that plays an important role in the development of leishmaniasis clinical manifestations. Plant-derived natural products have been recognized as bioactive agents with several properties, including anti-protozoal and anti-inflammatory activities. The present review focuses on the antileishmanial activity of plant-derived natural products that are able to modulate the inflammatory response in vitro and in vivo. The capability of crude extracts and some isolated substances in promoting an anti-inflammatory response during Leishmania infection may be used as part of an effective strategy to fight the disease.

In vitro evaluation of (-)α-bisabolol as a promising agent against Leishmania amazonensis.

Current treatments for leishmaniasis present some difficulties due to their toxicity, the use of the intravenous route for administration and therapy duration, which may lead to treatment discontinuation. The aim of this study is to investigate new treatment alternatives to improve patients well being. Therefore, we evaluated the inhibitory effect of (-)α-bisabolol, a sesquiterpene alcohol found in various essential oils of different plant species, against the promastigotes and intracellular amastigotes forms of Leishmania amazonensis, as well as the cytotoxic, morphological and ultrastructural alterations of treated cells. Promastigotes forms of L. amazonensis were incubated with (-)α-bisabolol to determine the antileishmanial activity of this compound. The cytotoxicity effect was evaluated by testing against J774.G8 cells. After these tests, the infected and uninfected cells with L. amazonensis were used to determine if the (-)α-bisabolol was able to kill intracellular parasites and to cause some morphological changes in the cells. The (-)α-bisabolol compound showed significant antileishmanial activity against promastigotes with a 50% effective concentration of 8.07 µg/ml (24 h) and 4.26 µg/ml (48 h). Against intracellular amastigotes the IC50 (inhibitory concentration) of (-)α-bisabolol (24 h) was 4.15 µg/ml. The (-)α-bisabolol also showed a cytotoxic effect against the macrophage strain J774.G8. The value of 50% cytotoxic concentration was 14.82 µg/ml showing that (-)α-bisabolol is less toxic to macrophages than to the parasite. Ultrastructural studies of treated promastigotes and amastigotes showed several alterations, such as loss of cytoplasmic organelles, including the nucleus, and the presence of lipid inclusions. This study showed that (-)α-bisabolol has promising antileishmanial properties, as it can act against the promastigote forms and is able to penetrate the cell, and is also active against the amastigote forms. About 69% of the promastigotes forms suffered mitochondrial membrane damage after treatment with IC50 of (-)α-bisabolol, suggesting inhibition of the metabolic activity of parasites. These results open new prospects for research that can contribute to the development of products based on essential oils or isolated compounds from plants for the treatment of cutaneous leishmaniasis.

Chemical and biological analyses of the essential oils and main constituents of Piper species.

The essential oils obtained from leaves of Piper duckei and Piper demeraranum by hydrodistillation were analyzed by gas chromatography-mass spectrometry. The main constituents found in P. demeraranum oil were limonene (19.3%) and ß-elemene (33.1%) and in P. duckei oil the major components found were germacrene D (14.7%) and trans-caryophyllene (27.1%). P. demeraranum and P. duckei oils exhibited biological activity, with IC(50) values between 15 to 76 µg mL(-1) against two Leishmania species, P. duckei oil being the most active. The cytotoxicity of the essential oils on mice peritoneal macrophage cells was insignificant, compared with the toxicity of pentamidine. The main mono- and sesquiterpene, limonene (IC(50) = 278 µM) and caryophyllene (IC(50) = 96 µM), were tested against the strains of Leishmania amazonensis, and the IC(50) values of these compounds were lower than those found for the essential oils of the Piper species. The HET-CAM test was used to evaluate the irritation potential of these oils as topical products, showing that these oils can be used as auxiliary medication in cases of cutaneous leishmaniasis, with less side effects and lower costs.

Composição química e atividade biológica dos óleos essenciais das folhas e caules de Rhodostemonodaphne parvifolia Madriñán (Lauraceae) / Composition and biology activity of essential oils from leaves and stems of Rhodostemonodaphne parvifolia Madriñán (Lauraceae)

Os óleos essenciais das folhas e caules de Rhodostemonodaphne parvifolia coletada na Reserva Florestal Adolpho Ducke foram obtidos por hidrodestilação e analisados por cromatografia em fase gasosa com detector de ionização de chama e espectrometria de massas. Treze constituintes foram identificados nas folhas, dos quais o ß-cariofileno foi o que apresentou maior percentual (41,30%). Nos caules foram identificados quinze compostos, com predominância dos sesquiterpenos ß-cariofileno (16,20%) e epi-cedrol (13,30%) e do monoterpeno linalol (15,40%). Os óleos foram analisados em ensaios químicos, de sequestro do radical livre DPPH, e enzimáticos, de inibição da enzima acetilcolinesterase. Nos ensaios de sequestro de radicais livres, apesar de terem se mostrado ativos em ensaios qualitativos, os óleos essenciais apresentaram fraca atividade antioxidante no ensaio quantitativo (inativos em concentração inferior a 1.000 µg/mL). A inibição da acetilcolinesterase só foi observada nos óleos essenciais das folhas. A composição química e a atividade biológica destes óleos estão sendo relatadas pela primeira vez neste trabalho.

The essential oils of leaves and stems of Rhodostemonodaphne parvifolia collected in the Adolpho Ducke Forest Reserve were obtained by hydrodistillation and analyzed by GC-FID and GC-MS. Thirteen components were identified in the leaf oil, of which ß-caryophyllene was the major (41.30%). Fifty components were identified in the stem oil, among them the sesquiterpenes ß-caryophyllene (16.20%) and epi-cedrol (13.30%), together with the monoterpene linalool (15.40%). The essential oils were analyzed at chemical, DPPH free radical scavenging; and enzymatic bioassays, acetylcholinesterase inhibition. At antioxidant DPPH assays, the oils showed to be active at the qualitative assay but only weakly active at quantitative test (inactive at concentration lower than 1,000 µg/mL). The inhibition of acetylcholinesterase was detected only in leaves essential oils. This paper describes for the first time the composition of these essential oils and their biological activities.

Quantitative determination by HPLC of iridoids in the bark and latex of himatanthus sucuuba

The main iridoids from the bark and latex of Himatanthus sucuuba were isolated and characterised by spectroscopic methods. HPLC was used for the quantitative analyses of these iridoids and the chromatograms of bark and latex showed a similar iridoid composition. Both parts of the plant are used in folk medicine for the treatment of various ailments.

Os iridóides principais das cascas e látex de Himatanthus sucuuba foram isolados e caracterizados por métodos espectroscópicos. CLAE foi utilizada para a análise quantitativa desses iridóides e os cromatogramas da casca e do látex apresentaram uma composição semelhante de iridóides. Ambas as partes da planta são utilizadas na medicina popular para o tratamento de várias doenças.